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Juniper Publishers-Open Access Journal of Case Studies

A Case Study on Birth Experience

Authored by Ayman Aboda

Abstract

Background: Normal consciousness may be profoundly changed during labour. Access to heightened states of personal and existential awareness may be part of the uninhibited ritual of natural childbirth.

Aim: Validate the possibility of heightened awareness and transcendence that can occur beyond the province of normal human experience. Peek experiences may bring life changing realizations of deep universal connection and personal insight.

Findings: Breakthrough experiences may occur in any state of intense physical focus, including the mindfulness of supported birth practices.

Discussion: The perspective reiterates the importance of rational, individualized, woman-centred care in birth.

Conclusion: The environment of birth, the safety and support of known and trusted care givers can allow birth events to unfold with the least possible intrusion. A hitherto under-recognized sequalae for such freedom may be the possibility of connecting to a state of expanded awareness that can bring profound unveiling of inner being and self-acceptance.

Keywords: Normal consciousness; Natural childbirth; Woman's emotional experience; Bleeding; Fearfulness; Birth experience; Acute medical resuscitation, Neurosurgeon

Statement of Significance

Problem

Constrictive models of care during birth.

What is already known

Current institutionalized models are driven by risk and litigation. Success in mitigating such harms has been tempered by spiralling costs of intervention and more insidiously, a diminution of the birth experience.

What this paper adds

The rationalization of appropriate care. The importance of choice and the understanding that birth can yield of itself, a glimpse into the extraordinary, an experience of wonder and life changing self-awareness that can be truly transformative for the mother and for all those with whom she shares her journey.

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Pushed to the Limit - Reclaiming the Ecstasy of Birth

“I remember when my world changed forever. I experienced something miraculous when I gave birth, I gave birth to myself as a mother. During labour, I noticed the world around me change. The lights seemed dimmer and there was a silence in the room. I was conscious but it felt different. There was an incredible sense of calm. I could sense everything, the bed, the room, my breath, and heartbeat, my thoughts and feelings. Time stood still. I was no longer connected to the room. I was distant in a place I had never been before. I was totally removed from everything. There was a sense of floating without boundaries. It was so calm, peaceful. I remember looking down as I hovered over my body, watching myself from above, as if I were two separate beings. I felt so loved. Fearless. I have never felt like this before, the beauty, the purity and bliss. A realisation of my existence came. It was as if I had gained clarity and the answer to life's many questions. Everything was exposed in its truest form. I knew why I was here and what I was born to do. I was here to create.”

GS – personal birth story.

What’s it like?

Long before the infamy of Covid 19, a philosopher once asked what it would be like to be a Bat? [1] Would it think as we do? Would it experience the world in the same way? Of course, we imagine that it would, but then all we’re really doing is putting ourselves within, thinking of it as just a smaller version of us; something that sees as we do, hears in the same way – albeit with bigger ears, and otherwise makes sense of its perceptions with the same quality and perspective of mind. But no one can really say for sure – which was in fact the whole point of the question: to remind us that all we can ever truly know is what it’s like to be us. And what is that you might then ask. Well, that’s easy, it’s the awareness we have of living, of being here, experiencing the world around us and knowing where we are within it. Now whilst this may or may not be true for other creatures - including bats, the outward-looking, body-centred perspective through which we frame our experience of living seems so much a given norm, that it’s hard to imagine there could be any other way of doing things. But is there?

Looking beyond the veil

In his book, “Life after Life”, Raymond Moody described a near-death experience which was noted following an acute medical resuscitation [2]. The survivor told his rescuers that whilst they’d been busy trying to save him, he’d been looking down as things occurred, watching the entire scene unfold from a point that was from somewhere outside his body. On the one hand he knew that this was something that shouldn’t normally happen and yet it had seemed so undeniably real that he couldn’t believe it hadn’t. More recently, American neurosurgeon Eban Alexander reported a similar sensorial shift with translocation of living awareness away from the body at a time when deep, septic coma had stopped all measurable brain activity [3].

These and other accounts suggest that the seat of consciousness may not be as firmly anchored as previously imagined, and that the usual, body-centred perspective may be just one of many possibilities available to us. But of course, we know this already for in our dreams each night, most of us experience vast multiplicities of non-ordinary awareness or altered states of being that we simply take for granted as flights of fantasy. In his book, “Journeys out of the Body,” Robert Monroe described specific techniques to intentionally induce similar states of transpersonal, or Out-of-Body Experience (OBE) in daytime too, where consciousness could exist replete and fully visceral despite separation from the physical body [4]. Cognitive awareness of self and surroundings remain corporally vital, with the experience most often described as a heightened or more splendid state of normal which could at times lead to an unprecedented feeling of bliss or profound belonging as if having become intimately connected to some greater universal presence or unconditional ease.

Not surprisingly, Out of Body Experiences can transform lives, and indeed they do. For the prophet Elija, the apostle Paul, Saint Francis of Assisi, the Buddha, and the modern spiritual teacher Ekhart Tolle, this was powerfully so. Sadly however, such revelations seem sparsely scattered, contained if lucky, to the zeal of a few determined practitioners or else the fervour fuelled of religious rapture or tribal ceremony such as the dreamtime of indigenous Ngangkari or shamanic ritual or more illicitly, from the psychedelic haze of contemporary drug culture. Without the agency of such provocateurs, one might reasonably wonder if the phenomenon was indeed meant to happen in normal life. Afterall, nature tends only to preserve or promulgate those behaviours that best suit the ability to survive - including the way we perceive the world. The paucity of OBE’s, notwithstanding their reputed glimpse of wonder, may simply reflect that they are less advantageous at keeping us alive - unless of course something changes that makes them more able to do so.

Holocaust survivor Viktor Frankl described looking down at his body during the terror of war and internment [5]. He thus felt distanced from the inhumanity and outrage of the horrors inflicted upon him and was somehow more able to survive. To withdraw from the immediacy and acuity of the senses can help to endure, to sustain trials of physical hardship and duress. Similarly, to quell the plethora of the senses, to focus intently may optimize performance in peek states and zones of intense concentration allowing an almost trance like state of deep attention for what needs to be done. In such states, the extraordinary can become possible and feelings of deep, transpersonal connection and grounding can arise.

Pushing to the limits –the energy of childbirth

“In the most intense hours of labour, I became mindless, floating in boundless space between contractions. All thoughts receded. My mind plummeted into an immense silence bathed in love and well-being. I felt a oneness with all mothers who had ever birthed” [6].

A woman's emotional experience during labour may show common themes, beginning with excitement and a need to feel safe and supported as her contractions develop which means she can then focus her energy on getting through things, one surge at a time [7]. Her world then narrows as concentration deepens and she may become less aware of things around her, moving into a ‘zone’ of timelessness, of dissociation or disconnection with the rest of the world. In this phase, perhaps 10% of women may experience states of altered awareness, including fully-fledged OBE’s where the boundaries of self and ordinary reality become less defined and deeper, more subtle levels of creativity and spiritual encounter can occur. These feelings then transition and become more grounded as an urge to push strengthens and a reconnection with the present emerges, often bringing a realization of awe and amazement as the power of their bodies moves them irresistibly towards birth [8].

modelling

“In achieving the safety of childbirth our society may have lost more than it has gained.” [9]

Birth however is not always perfect. Whilst it is a natural process, left to itself, it may not always lead towards a desired or safe ending. There may be pain, unprecedented in its intensity; bleeding, a fearfulness of what lies ahead, of what remains unknown or unexpected. There may be escalating risk or concern that must be managed. Models of care have thus evolved to mitigate, to wrestle control from such uncertainty, to bring order to chaos, to reign-in the freedoms of nature’s way. And so, we change. We change things, and more sadly so, we change ourselves. We stop seeing women as mothers or as mothers-to-be; they become patients. We take them from home, from community and on-country, and admit them to hospital to be cared for, to be monitored, to be made right. Labour becomes a diagnosis, an illness, a condition that must be treated and fixed. Natural rhythms are ignored and give way to ever more intrusive cascades of intervention.

Without doubt, neonatal and maternal outcomes have improved, but there comes too a growing apprehension of cost, of over-service, and more distressingly, an ever-widening wound of visceral and emotional discontent that follows thereafter. What began in good faith, what began because we genuinely care, because we wanted to help, has become a mandate. We now do what we do because we feel we have to. We feel we must. We believe it’s best. But is it always so?

By institutionalizing the birth experience, we can diminish a woman’s ability to participate, to help nurture the power and flow of her body to progress naturally and spontaneously. More subtly, by controlling her experience so arduously, we hide from her the opportunity to encounter an intimate, perhaps sacred awakening of deeper self that might otherwise have occurred. We thus sterilise the possibility of transience, of enchantment. We diminish the extraordinary to something best forgotten. We brush it aside as vagrant, a symptom of momentary dysfunction, or the rekindling of aberrant memory in the fever of fatigue [10].

Pushing back - reclaiming safe passage

Birth is of itself complete, it lacks nothing to make it more so. We have tried to make it better and of course in many ways we have, but at what cost? And for such gains won, how much has been lost? In our desire to make it fit, to conform to modern standards, there may be much that we have taken, that we have forsaken. We may have stolen from it something subtle, an opportunity to transcend normal living and experience an intimate revelation of profound connection and wonder.

The awareness we have of ourselves in the physical world is miraculous, a perspective made possible by the wondrous workings of what makes us human. Occasionally however, states of expanded, other-worldly consciousness can occur, where glimpses of a far-from-normal reality can pervade our senses with life changing ardour. When we consider the impact that such transformative experiences can bring, we may well grieve for their paucity.

In Australia, over 300 000 women give birth annually. 1 in 10 of these may experience feelings of altered consciousness which for some, may lead to an encounter of unprecedented personal realization and bliss. Imagine the impact that such life-changing experiences could bring. Imagine a birthing space where the unfolding of such deep ritual was nurtured, where we protect and gently support each mother to follow if she desires, the natural rhythms of her body. Where we gave freedom to experience the power of birth, and, if it should happen, the revelation of divine inner being that may enrich her life thereafter. Imagine what this would bring for her little one as she bonds and for those with whom her life is shared. Imagine what this would do to her world. To yours. To ours. It’s not just a want, it’s a rite worth wanting, a rite worth reclaiming. All we need do, is just let it happen. I think we can.

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Effect of Different Water Sources on Survival Rate (%) Growth Performance, Feed Utilization, Fish Yield, and Economic Evaluation on Nile Tilapia (Oreochromis niloticus) Monosex Reared in Earthen Ponds- Juniper Publishers

Abstract

The aim of the present study was to investigate the effect of water source on survival rate %, growth performance, feed utilization, fish yield, economic evaluation and production of Nile tilapia (Oreochromis niloticus) monosex reared in earthen ponds. Nine earthen ponds were used and divided into three categories of three earthen ponds each. The average size of each pond was approximately 5200m2, 6000 monosex all male Nile tilapia were used in each pond and were stocked for 192 days. The fingerlings average weight was 4.38±0.03g/ fish, the fish were fed using a floating feed 25% crude protein, and were fed at a daily rate of 3% of their body weight. Results showed that body weight was increased significantly (P<0.05) with well water to 472.33g/fish. While were 354.17 and 320.17g/fish for fresh and agricultural drainage water, respectively. Specific growth rates (SGR%) increased with well water compared to both fresh and drainage water. Feed conversion ratio (FCR) and protein efficiency ratio (PER) were improved with Agriculture drainage water. Survival rates with fresh and well water were 98.53% and 98.31% respectively, however, was 95.05% with Agriculture drainage water. Total fish yield were affected significantly by treatments. It was 2128, 1921.8, and 2837.7kg at fresh, drainage and well water respectively. Net return arrived to 12996 for well water source when it was 6784LE for agricultural drainage water and 9158LE for fresh water.

Keywords: Water resources; Nile tilapia; Growth per

Introduction

Nile tilapia, Oreochromis niloticus (Linnaeus) is one of the most important freshwater fish in world aquaculture [1]. It is widely cultured in many tropical and subtropical countries of the world [2]. Rapid growth rates, high tolerance to adverse environmental conditions, efficient feed conversion, ease of spawning, resistance to disease and good consumer acceptance make it a suitable fish for culture [3]. Farmed tilapia production increased semi dramatically in recent years, increasing from 383,654mt in 1990 to 2,326,413mt in 2006 [4]. Tilapia has established a secure position in a number of water impoundments of India. But, its performance in open water ponds of the country has been discouraging over the years [5]. For tilapia aquaculture is excessive reproduction and the resulting small size of the fish produced.

Egypt has suitable natural conditions for desert aquaculture. Egypt has vast resources of groundwater [6]. Fresh groundwater resources in Egypt contribute 20% to the potential water resources in Egypt. One of the groundwater resources is the Nile Valley and Delta system with the storage capacities of 200 billion m3 and 300 billion m3, respectively. Oasis water in the west desert, Bahariya, Farafra, Dakhla, Kharga, and Siwa, were established from underground natural wells and springs.

With the prohibition of the establishment of fish farms on agricultural land, with the prohibition of the use of Nile water for fish farming, with increased competition for spaces adjacent to the lakes and sources of agricultural drainage water, despite its disadvantages, has caused the possession of new fish farm in the Nile [7]. Valley of the most difficult things and out of reach. Hence the search for an alternative to invade the desert, especially with the development of methods of fish farming and providing the requirements of education and with the provision of underground water of the highest purity with different salinity (fresh & brackish & marine) and where the trained professionals are available [8]. In the hope to produce clean fish with improved quality and cheaper than other animal proteins we conducted the present research in a private fish farm located in the desert belonging to Noubaria Agricultural Development company (Ragab Farms) aiming to study the effect of water source on survival rate (%) growth performances, feed conversion ratio, protein efficiency ratio, annul fish yield and profitability Nile tilapia (Oreochromis niloticus) monosex commercial farming.

Materials and Methods

Water Source

Three types of water sources: fresh water, agricultural drainage water, and well water were compared in the present experiment. Water supplies were replaced three times during the experimental period (192 days).

Experimental design

Nine earthen ponds (5200m2) were used in these experiment were divided into three categories of earthen ponds even three ponds represent one treatment (fresh water, drainage water and well water.

Stocking density

6000 monosex all male Nile tilapia (Oreochromis niloticus) fingerlings of average weight (4.38±0.03g/ fish) were stoked in each pond on April 11, 2007 and observed through October 19, 2007. The area of each pond 5200m2.

Experimental Fish

Fingerlings of all male Nile tilapia (Oreochromis niloticus) monosex were collected from Noubaria Agricultural Development Company (Ragab Fish Hatchery) and were over wintered in earthen ponds to provide suitable fingerlings for the beginning of the growing season. All ponds in this experiment were sampled monthly using a cast net method. Sample sizes were 1% of the stocked numbers and the average individual fish weight was calculated to determine growth rates. Then, with these calculations, the feed amounts were adjusted for the following month.

Experimental diet

The floating commercial diet used in this experiment was fed at a daily rate of 3% of the fish body weight by using self feeders The ingredients of the commercial diet used in the experiment is presented in (Table 1). The dietary composition of vitamin and mineral premix is listed in. Fish were fed a floating ration for 6 days per week. Feeding rate was adjusted monthly based upon the calculated biomass of fish obtained through the monthly sampling and assumption of 100% survival.

Water quality

Physical parameters: Water temperature °C was determined at every days in the experiment.

Chemical Parameters: Samples for determination of dissolved oxygen (DO) were immediately fixed after sampling and DO concentration was determined according to Winkler's technique. Methods described by Golterman et al. [9] were used in determination of ammonia. Also pH was measured by digital pH meter (Orion model 720 A, s /No 13602) in all experiments.

Chemical Analysis of the commercial of Diet: Chemical analysis of the commercial diet used in the experiment was done according to AOAC (2000) as shown in Table 1.

Growth parameters and Statistical analysis: Data on growth, feed utilization, survival rate and proximate and chemical composition of whole fish body were subjected to one-way ANOVA [10]. To locate significant differences between fish size within different water resources of pond. Duncan's multiple rang test [11] was done. All percentages and ratio were transformed to arcsine values prior to analysis [12].

Results and Discussion

Experimental diet

The commercial diet used in the present experiment contained 25% CP and 4.3kcal/g gross energy (Table 1). Although there are large variations in the data available about the optimum protein level for tilapias which range between 20 and 40% crude protein [13-15] practical diets as low as 25% protein was successfully used for rearing monosex tilapia [3].

Vit. A 8000 I.U. Vit. D3 4000 I.U.; vit. E 50mg; Vit. k3 19mg;

Vit. B1 40mg; vit. B2 25 mg; Vit. B6 125mg; vit B12 69mg;

Pantothenic acid 40mg; Nicotinic acid 125mg; Folic acid 400mg;

Water quality

Collected data on water temperature and dissolved oxygen (DO), pH and ammonia are summarized in Tables 2-4. Water temperature throughout the present experiments ranged between 24.13±0.53 and 30.26±0.45 °C in fresh water experiment, 24.23±0.53 and 30.65±0.53 °C in drainage water experiment and between 29.94±0.12 and 33.63±0.43 in well water experiment which was the high temperature and closely related to the average of optimal value for tilapia (28-30 °C). Our results were agreement with Broussard [16] reported that tilapia as a warm water fish that dominate African lakes, are known to grow well in high temperature. The fluctuation of water temperature are reached its maximum values during August, however its minimum were during April and November.

*Each value was on average of four sub samples

Biotin 20mg; cholin chloride 80 mg; copper 400mg; Iodine 40mg;

Iron 120mg; Manganese 220mg; Zink 22mg; Selenium 4mg

Means in the same column having different letters are significantly different (P<0.05).

Overall means of water dissolved oxygen (DO) throughout the present experiment were 7.20±0.37mg DO/I for fresh water, 7.19±0.36mg DO/I for drainage water and 6.33±0.36mg DO/I for well water. The fluctuation of water dissolved oxygen (DO) showed that the maximum values of DO were obtained in November for the fresh and drainage water and August in well water, however, the lowest values were in April. In general, dissolved oxygen levels were within the high standards and higher than cited by Boyd [18] for good production of tilapia (4.20 to 5.90mg DO/I) in aquaculture ponds. One of the most important environmental factors is dissolved oxygen. It is considered a limiting factor for success or failure in intensive culture. An excellent aquaculture attribute of tilapia is their tolerance to low dissolved oxygen concentration [16]. The dissolved oxygen content in earthen ponds depends on the pond water temperature, fish biomass and rate of water exchange [18]. Chervinski [19] reported that O. niloticus survived short term exposure to 0.1mg DO/ l. However, Collins [20] observed in a review on oxygen concentration of various studies, that growth rate of non-salmonid fish was increasingly depressed as dissolved oxygen fall below 50% saturation. Rappaport et al. [21] reported that growth of carp was reduced by predawn dissolved oxygen less that 25% saturation. Tichert-C & Green [22] compared the growth of tilapia monosex in earthen ponds aerated or unaerated at 10 or 30% saturation of dissolved oxygen. They found that tilapia production and final weight were significantly greater in aerated ponds than unaerated ponds.

The water pH values throughout the present experiments ranged between 8.00±0.13and 8.10±0.13 with an overall mean of 8.04±0.13 in fresh water and ranged between 8.01±0.13 and 8.10±0.13 with an overall mean of 8.05±0.13 in drainage water and ranged between 7.98±0.13and 8.01±0.13 with an overall mean of 8.00±0.13 in well water. The fluctuations of pH reach the highest value of 8.10+0.13 during August in fresh and drainage water and 8.01+0.13 in well water. The results showed that the present pH values are suitable. For rearing tilapia monosex in earthen ponds. Johnson [23] recommended the range of pH 6.5 to 9.0 for most of freshwater fish species.

The water un-ionized ammonia (NH3) throughout the present experiments ranged between 0.09±0.01 and 0.12±0.01 with an overall mean of 0.11±0.01 in fresh water and ranged between 0.10±0.01 and 0.13±0.01 with an overall mean of 0.11±0.01 in drainage water and ranged between 0.06±0.01 and 0.10±0.01 with an overall mean of 0.077±0.01 in well water. The fluctuations of un-ionized ammonia reach the highest values of 0.13mg/ l during August. Unionized ammonia concentrations in the experimental ponds generally remained below levels which would cause chronic toxicity problems in tilapia. Tilapia is more tolerant to elevated levels of ammonia than more other sensitive species such as salmonids [23]. Some tilapias have been shown to acclimate to higher levels of ammonia after chronic exposure to low levels [24]. Johnson [23] showed that levels of un-ionized ammonia which may adversely affect growth in tilapia range from 1mg/ l to 2mg/ l ammonia where temperature and pH are within normal range.

Growth performance of tilapia monosex

Mean weight: Results of the present study showed that the mean weights at all rearing intervals different significantly (P<0.05) during all the experimental periods (Table 5 & Figure 1). Averages of fish body weights for fresh water, drainage water and well water were found to be 23.16, 18.66 and 25.16g, respectively after the 1st month of stocking. The statistical evaluation of results indicated that live weights at this period increased significantly (P<0.05) with using well water. A similar trend was also observed in fish body weights during the other growing periods. At harvest average body weight of fish stocked at well water was significantly (P<0.05) higher than that of fish stocked in fresh or drainage water, which indicates that weights fish were decreased in fresh and drainage water with increasing for used well water at harvest were 354.17g, 320.17g and 472.33g for fresh, drainage and well water, respectively. This significant advancement in fish body weights with increasing at higher temperature of water advocated by Azaze et al. [25] reported that the final mean weight was significantly higher at 26 and 30 °C than at 22 and 34 °C. This finding agrees with our results.

Average daily gain (ADG g/day): Results presented in Table 5 revealed that water sources, affected significantly (P<0.05) ADG during all experimental periods tested (30, 60, 90.120.150.180 and 210 days after start). In general these results indicated that the well water favored significantly ADG of the tilapia monosex in intensive culture system. The results of this point were in agreement with those found by [17] who grew O. niloticus from 49g to 271g in 122 days (1.4%/day). Siddiqui et al. [15] found that ADG of tilapia O. niloticus reared for 98 days at different water exchange in outdoor concrete tanks was 1.06g / day at 30% dietary crude protein. In the present study the average daily gain was higher with 25% crude protein at all treatments. However, the optimal feeding rate depends on fish size and Specific Growth Rate (SGR %): Results presented in Table 5, revealed that water sources, affected significantly (P<0.05) SGR% during all experimental periods tested (30, 60, 90.120.150.180 and 205 days after start). In general these results indicated that the well water, favored significantly (P<0.05) SGR% of the tilapia monosex in intensive culture system.

During all tested experimental periods tested (30, 60, 90.120.150.180 and 205 days after start) SGR% increased significantly (P<0.05) in almost linear manner in the well water than fresh and drainage water In the present study SGR% values in case of well water continuously higher than fresh or drainage water in all experimental periods. This may be due to the higher temperature of the well water (average 31.94 °C) compared to 27.47 and 27.81 °C for fresh and drainage water, respectively. The results obtained in SGR% are in agreement with those found by Eid & El Denasoury [27] who indicated that increasing temperature from 16 °C to 27 °C improve growth rate of Nile tilapia, which using well water.

Feed conversion ratio (FCR): Results presented in Table 5, show that there were significant (P<0.05) effects of water sources on FCR, feed conversion ratio was observed at harvest was 2.87 at fresh water, followed by 2.83 at well water and 2.80 at drainage water and 2.94 for 1700m2 followed by 2.89 for 4000m2 followed by 2.75 for 5200m2 and was 2.57 for 6000 fish/ acre, 2.75 for 8000 fish/ acre and 2.78 for 10000 fish/ acre. The analyses of variance of the FCR values are presented in Table 5. The FCR is affected by the physiological state of the fish, environmental condition, [28]. Lovshin et al. [29] found that FCR for all male tilapia in earthen ponds was higher (4.3) than when compared with all male and female tilapia in earthen ponds (FCR=7.2). while, fish growth is affected by the amount of feed consumed and the efficiency of assimilation [30].

Protein efficiency ratio (PER): Results of protein efficiency ratio (PER) are presented in Table 5, There were significant (P<0.05) effects of water sources, on PER, it improved significantly (P<0.05) with each increase in pond sizes and decrease stocking density throughout the experimental periods. The best PER observed at harvest was 1.42 with drainage water, followed by 1.40 at well water and 1.38 at fresh water Nyina-W et al. [31] confirmed that when protein supply is appropriate (400500g protein/kg feed for percid fish), different lipid contents in feeds do not have an impact on the rearing results of pikeperch.

Fish survival rate: Results in Table 6 showed that survival rates were changed significantly (P<0.05) by water resources, in fresh and well water were insignificantly (P<0.05) different but survival of the fish in drainage water was 95.05% which was less than survival rates in both fresh water and well water indicating the probable effect of some faction of water quality.

Fish yield: Results of Table 4 show fish yield (kg) per acre as affected by water sources,. Results revealed that total yield increased significantly (P<0.05) with well water. The total production was found to be 133.34% and 90.31% for well water and drainage water, respectively, while it was found to be 76.33% and 68.84% for 4000m2 and 5200m2.

The results of the present experiment were similar to those of Tal & Ziv [32] who showed that the net yield of tilapia monosex in earthen ponds was 16750Kg /ha (7035.0kg/ acre) after 100 days of stocking of 80.000 fish/ha, (33600 fish/acre, 8 fish/m2) on the other hand Eid & Denasoury [27] indicated that increasing temperature from 16 °C to 27 °C improved growth rate of Nile tilapia. Watanabe et al. [33] found that growth rates generally increase with increasing temperature and where markedly lower at 22 °C and well water is the best because the temperature constant through the year and the best quality of the water. [34] found the higher yield obtained in small pond sizes because the bigger ponds with greater surface area were more difficult to manage and often resulted in lower fish yields.

All fish species are characterized by an ideal range of temperature in which they show their maximum growth [3537]. Several studies have been reported that the specific water temperature range showed the faster growth in Pikeperch, Sander lucioperca at 20 °C to 25 °C [38-40]. Low temperature causes sluggishness by retarding the digestion speeding of fish [41]. Some researchers have found that the digestion rate has been increased as the temperature increases [42]. Environmental temperature is one of the most important ecological factor which also influence the behavior and physiological process of aquatic animals [43].

One of the major advantages of groundwater sources is their constant temperature throughout the year. Shallow sources of groundwater approximate the mean air temperature of the area. The chemistry of groundwater is directly dependent on the geology of the area surrounding the source. In limestone areas, groundwater is hard, and high in calcium and carbon dioxide [44]. In areas of granite formation, the groundwater tends to be soft, low in dissolved minerals and carbon dioxide. As will be discussed later, there are advantages and disadvantages to both, emphasizing the need for early extensive water quality testing.

Water temperature has substantial effect on fish metabolism. In response to decreasing of water temperature the enzyme activity of tissues have been increased [45]. Velmurugan et al. [46] have investigated that histopathological and tissue enzyme changes of C. gariepinus exposed to nitrite when water temperatures changes from 27 °C to 35 °C. In a stressful and unfavorable environmental condition GPT and GOT may increase in blood serum. In the present study serum GPT and GOT level were affected by different water temperature. Serum GPT and GOT amount in different fish fed at 20 °C are comparatively lower than those of fish fed at 16 °C and 24 °C experiments (Tables 1-3). These results indicated that 20 °C may be a favorable water temperature for better growth of 16g juvenile Korean rockfish [47,48].

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Quality by Design in Enzyme Catalyzed Reactions-JuniperPublishers

Journal of Chemistry-JuniperPublishers

                                Abstract

Quality by Design is the new-age path chosen towards achieving the demanding quality standards in pharmaceutical industry. The present paper aims to throw light on Pharmaceutical Quality byDesign (QbD) and how its implementation will help manufacture better quality of Pharmaceuticals. Quality by Design is introduced along with its key elements to help make the understanding process easier. To attain built-in quality is the primary objective of Quality by Design. Finally, it can be said that the quality that is achieved by end product testing is not something that can be guaranteed unlike the quality assurance that can be provided by Quality by Design.

Keywords: Quality by Design (QbD); Quality Target Product Profile; Design Space; Critical Quality Attributes

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Introduction

“Quality Can Be Planned.”-Joseph Juran

The above quote is self-explanatory when it comes to product quality in the pharmaceutical manufacturing industry. Quality by design (QbD) is not very old but a recent inclusion in the pharmaceutical industry. It`s sole objective is to achieve better quality standards that is especially important in the pharmaceutical industry. The QbD approach consists of various components, important ones being risk assessment, assessment and management of the identified risks, design of experiments (DoE), quality target product profile (QTPP), and establishing a control strategy to keep the product within the design space that was created with the QbD study [1]. Out of all the components, a lot of pharmaceutical development studies have incorporated DoE for a more rational approach [2].

The target of analytical QbD approach is to establish a design space (DS) of critical process parameters (CPPs) where the critical quality attributes (CQAs) of the method have been assured to fulfil the desired requirements with a selected probability [3-4].

The principles that are involved in the pharmaceutical development and are relevant to QbD are all described in the ICH guidelines (ICHQ8-11) [5].

Any Pharmaceutical Development Process Typically Covers the Following Sections:

a) Complete portfolio including all the details as well as analysis of the Reference Listed Drug Product

b) Quality Target Product Profile (QTTP) compilation.

c) Figuring out the Critical Quality Attributes (CQA)

d) Complete characterization of API &CMA (Components of Drug product) identification of the API

e) Excipient selection& excipients CMA identification

f) Formulation Development

g) Manufacturing Process Development [6]

Quality Target Product Profile (QTPP) describes the design criteria for the product, and should therefore form the basis for development of the CQAs, CPPs, and control strategy.

Critical Quality Attributes (CQA) – A physical, chemical, biological, or microbiological property or characteristic that should be within an appropriate limit, range, or distribution to ensure the desired product quality (ICH Q8) Critical Process Parameter (CPP) – A process parameter whose variability has an impact on a CQA and therefore should be monitored or controlled to ensure the process produces the desired quality. (ICH Q8) Critical Process Parameters (CPP) identification and their impact analysis is done by conducting a preliminary risk analysis for every process parameter (PP) that is involved in the individual unit operations.

Need for QbD in Pharmaceutical Industry [7,8]:

a) To integratepatient needs, quality requirements and scientific knowledge all in one design while the pharmaceutical product is still under developmentand further extending to the manufacturing process.

b) To have a better understanding about the impact of raw materials and process parameters on the quality of the final product. This is especially important for biopharmaceutical products since raw materials like cell culture media can be the risk for variability, effecting important factors likecellular viability, cell growth and specific productivity.

c) To collaborate closely with rest of the industries and the regulators and successfully keep up with the regulatory reviews

d) To maintain harmonization in all the regions so that a single CMC submission worldwide is all that is needed.

e) To encourage continuous quality improvement for the benefit of patients.

f) To enable better product design that will have less problems while manufacturing, thus facilitating more efficiency in the manufacturing process.

g) To make post-approval changes easier since it will be contained within a pre-defined design space, thus resulting in regulatory flexibility.

Every production process in a pharmaceutical industry to implement certain control strategies with the ultimate goal of a robust process. A robust process is the gateway to high product quality at the end of the day [9]. Process variability stands as a hurdle to process robustness, and this originates from lack of control on the process parameters. Thus, QbD steps-in to avoidbatch to batch variability in pharmaceutical products [10].

The net outcome of the detailed QbD study (applied in any product) is the segregation of process parameters with respect to their criticality and the finalization of a proven acceptable range (PAR) for every operation. The knowledge that is gained post the QbD evaluation encompasses every minute detail of the operational process as well as the product in general, and lead to the defining of a Design Space. This way, the impact that the manufacturing process might have with regard to the variability of the CQAs becomes apparent, which helps in strategizing testing, quality and monitoring of batches [11].

Process Evaluation: Linking Process Parameters to Quality Attributes

It is important to carefully evaluate the process completely before applying QbD to it. The better knowledge you have of the process, the more effective your QbD will be. Moreover, process characterization is required to specify the proven acceptable ranges (PAR) for critical process parameters (CPPs). In the traditional approach that is implemented in biopharmaceutical production, existing empirical process knowledge is used on a daily basis. However, this approach leads tolaborious and time consuming post approval changes during process adaptation and any new technology implementation that may have become necessary for raising the efficiency of the process. Also, the effects aprocess scale-up can have on the quality of the final product cannot be predicted when using the empirical process development.

This can increase costs and also can cause difficulty in implementing any changes in the set manufacturing process. Thus was born a way to achieve deeper understanding of processes which would lead to greater flexibility and freedom to effect changes. The concept of operation under a pre-defined design space gave this flexibility. Design space is nothing but a concept that is a part of the “Quality by Design” (QbD) paradigm. Now, manufacturers are to follow a science-based process development than their empirical counterpart.

The QbD Concept is Best Explained in this Flowchart Below

Define a Quality Target Product Profile (QTPP) for product performance

⇩

Identify its Critical Quality Attributes (CQAs)

⇩

Create experimental design (DoE)

⇩

Analysis done to understand the impact of Critical Process Parameters (CPP) on CQAs

⇩

Identify and control the sources of variability.

Process characterization sets the ball rolling in any process development, which employs a sound risk assessment rating the various critical process parameters according to their importance [12-14].

Downstream Processing in Biotransformation

Downstream processes of biopharmaceutical industry essentially include the following steps:

a) Harvesting

b) Isolation

c) Purification

Various unit operations that constitute any biopharmaceutical process follow a designed sequence to form an integrated process [15]. Thus, any change in any one of the one-unit operation can affect the functioning of the subsequent unit operations. This is the reason why interaction effects between participating parameters across unit operations should also be taken into account during the process development. Interactions are said to happen when setting of a parameter will show effect on the response of another parameter. Due to this dependence between the parameters, the combined effects of any two parameters hailing from different unit operations cannot be predicted from their individual effects. Regulatory authorities demand inclusion of interactions of parameters within the QbD approach during any process optimization [16]

Example: Downstream processing of 1, 3-propanediol

Process: Fermentation

Fermentation broth that uses flocculation, reactive extraction, and distillation was studied. Flocculation of soluble protein as well as cellular debris that were present in the broth was carried out by using optimal concentrations of chitosan (150 ppm) and polyacrylamide (70 ppm). It was seen that the soluble protein that was present in the broth decreased to 0.06 g L-1. Recovery ratio (supernatant liquor: broth was found to be greater than 99% (Figure 1) [17,18].

The above flowchart shows a typical fermentation process broken down in steps. Glycerol fermentation process is taken as example for the illustration [19].

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Case Study for API

API product development from the very nascent stages require a lot of planning when implementing QbD at every stage. Whether it is two-step process or a multi-step process, each and every operation and parameter needs to be scrutinized before creating a relevant design space. Brainstorming every possible roadblock that might threaten the quality of the final pharma product is what will help design a top-quality process. A futuristic vision is important in the initial steps of QbD planning. The most important part is to pay sufficient attention to detail lest critical aspects might be missed. This is best done by sitting with the entire development team and taking every minor detail into account. Given below is a case study for a API intermediate development process with the help of QbD that highlights the important steps as to how to go about implementing it from the very beginning of your research. QbD is done best, when it is implemented from the very nascent stage of product development.

Quality Target Product Profile

When making your QTPP, make sure you list down everything from your vendor details to target costing. This step basically asks you to think of every aspect of your product and make a comprehensive profile of it. The specification of quality must be highlighted here with all the challenging impurities that might threaten your quality. Everything from stability testing requirements to raw material quality [20] is encompassed in this stage of QbD.

CQA Determination

Given below are some typical CQA parameters that are considered in most of the enzymatic methods of API intermediate preparations.

a) Purity

b) Chiral purity

c) Enzyme residue

d) Assay

e) Appearance

f) Residual Solvent

g) Yield

h) Polymorphic forms

i) Moisture content

j) Melting point (Table 1)

Initial Risk Assessment

The risk assessment can be done in various ways and is the customizable step in QbD. This part calls for a group-discussion or a team meeting where everyone can list down all possible risks related to the project in discussion and grade each one in the list with the amount of risk that it poses. The simplest module suggests you number them 1, 2, 3 with the increasing or decreasing order of the risk threat. A more complicated and detailed risk assessment requires linking of CQAs and CPAs to highlight the risk of their interdependence (Figure 2) [21].

Post risk assessment, comes the control strategies to be followed to tackle the possible risks that are probable. The control strategies are for you to think and execute to achieve your target quality specifications.

Design of Experiment

This is a valuable tool for channelizing your experimental work, to move ahead in a systematic manner. Design of experiments can be of several types: comparative, screening, response surface modeling, and regression modeling [1].

Comparative Experiments: The aim of this study is simple, i.e., picking best out of two options. The selection can be done by the comparison data generated, which is the average of the sample of data.

Screening Experiments: If you want to zero-in on key factors affecting a response, screening experiments would be the best bet. For this, list down concise list of factors that might have critical effects on response that you desire. This model serves as preliminary analysis during development studies.

Response Surface Modeling: Once you have identified the critical factors that affect your desired response, response surface modeling comes handy to identify a target and/or minimize or maximize a response.

Regression Modeling: This is used to estimate the dependence of a response variable on the process inputs.

A step by step guide is given for the DoE step of the QbD process (Figure 3).

Response columns were filled post experimentation as per the design creation (Figure 4).

Factorial Design Analysis Done as Given Under

Analysis Done First for One of the Responses, “Yield”: (Table 2)

P-Values Were Checked for Significance and Higher P-Value Term Eliminated First to Create a Reduced Model:

(Table 3) (Figures 5 & 6)

Observation

From the above graph, significant interaction between the two terms can be inferred.

Analysis Done for the Response “Diacid”: (Table 4)

P-values Checked for Significance and Higher P-Value Term Eliminated First to Create a Reduced Model: (Table 5) (Figure 7)

Observation

From the above graph, significant interaction between the two terms can be inferred.

Response Optimizer Was Used to Optimize Both The Terms With Respective to The Given Responses- Yield and Diacid: (Table 6) (Figure 8)

The optimized parameters predicted for maximum yield and minimum impurity (of di-acid) was found to be 8pH and 37C.

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Case Study 2

As mentioned before, regression analysis is another important tool that can be used to study existing data. This means that if you have done some experiments (without designing them beforehand), you can quickly run a regression analysis of the collected data to derive a relationship between CPPs and the reaction results.

A lot of times, when one follows the one-factor-at-a-time optimization process, by the time any CPP is optimized, a lot of data stands generated. Instead of just tabulating the data and wasting time manually making sense out of them, regression analysis can come to your rescue. As always, graphical data representation seems much easier to understand and also saves your valuable time.

The effect of pH was studied [22] separately in the preparation of deoxynojirimycin base (stage III). The reaction involved N-formyl amino sorbitol, water, oxygen and whole cells of Gluconobacter oxydans DSM2003. Later involvement of sodium hydroxide and sodium borohydride gave rise to deoxynojirimycin. Further work-up and 2-methoxy ethanol facilitated crystallization yielded Deoxynojirimycin base. In this experiment, pH of the reaction was changed to find out its role during the reaction and a regression analysis was run using Minitab to study this affect.

Observations recorded showed that reaction did not occur at pH2 and at pH8, the reaction did not reach completion. The optimum pH range between 4 to 6 showed certain effect on yield and purity. The significance of pH variation during the reaction was thus established as described below (Graphs 1-3):

When null hypothesis p-test was carried out, no significant effect of pH was to be found on product purity, impurity1 and impurity2, but its significant influence was seen in minimizing impurity3.

Furthermore, large-scale batches conducted were statistically analyzed as well to achieve better understanding of the influence of list of parameters on the output obtained. The following parameters were studied during the stage III reaction described above:

a) pH, RPM and Oxygen cylinders consumed during the course of the reaction.

Their effect on the output and reaction completion time was studied. It was seen that only RPM showed statistically significant effect on the reaction completion time and rest of the factors did not contribute to any significant effect on the output or reaction completion time.

During biotransformation process, i.e. during oxidation of N-formyl using Gluconobacter oxydans DSM2003 whole cell, three main unknown impurities peaks were observed in HPLC chromatogram while reaction monitoring. This process is capable of removing these three impurities during down streaming, work up & isolation to the levels mentioned below:

a) Impurity 1 (has defined RRT on HPLC chromatogram) not more than 3%

b) Impurity 2 (any other unknown impurity) not more than 1%

c) Impurity 3 (has defined RRT on HPLC chromatogram) not more than 10 %

Since higher level of impurities affect the yield of the process, efforts were carried out to study the factors which can reduce the formation of process impurities.

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Conclusion

The concept of Quality by Design (QbD) is highly reliable when it comes to achieving foolproof quality of your product. This is a modern tool that is going viral in Pharmaceutical industry especially because this industry demands high quality standards and tolerates no compromise when it comes to the quality. Breaking down QbD, it essentially comes down to identifying the critical parameters of the process and assigning a particular design space for every single critical attribute. Thus, QbD can be considered as an intelligent approach to quality that yields robust processes. QbD also ensures that there is continuous improvement in the process during the entire lifecycle of a Pharmaceutical product [23].

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Acknowledgement

Our group would like to thank the Department of Scientific and Industrial Research India, Dr. Hari Babu (COO Mylan), Sanjeev Sethi (Chief Scientific Office Mylan Inc); Dr. Abhijit Deshmukh (Head of Global OSD Scientific Affairs); Dr. Yasir Rawjee {Head - Global API}, Dr. Sureshbabu Jayachandra (Head of Chemical Research); Dr. Suryanarayana Mulukutla (Head Analytical Dept MLL API R & D) as well as analytical development team of Mylan Laboratories Limited for their encouragement and support. We would also like to thank Dr. Narahari Ambati (AGC- India IP) & his Intellectual property team for their support.

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Let’s Talk and Grow Together: A Bidirectional Communication between Granulosa- and Oocyte Derived Factors in the Ovary

Abstract

Reproduction, one of the most active and appealing area of research for endocrinologists and reproductive biologists since many a years, has several faces that remains to be unmasked in terms of its regulatory aspects. Available information on the regulation of oocyte development and maturational competence are gaping and needs elucidation to achieve utmost quality of eggs, a major area of concern. The notion of the somatic follicular cells providing an appropriate microenvironment for the development of oocyte throughout its journey has been replaced with the current perception of a complex yet regulated cross-talk between the granulosa-and oocyte-derived factors to orchestrate follicle development. Interestingly, actions of FSH and LH are mediated or modulated by these locally produced non-steroidal peptide factors from the follicular layer and the oocyte itself (insulin-like growth factors (IGFs), epidermal growth factor (EGF) family members, TGFβ super family members etc.), forming an intimate regulatory network within the ovarian follicles. Present article will provide a deeper insight into the need and underlying mechanisms of action of these growth factors in the intraovarian network to sustain a healthy oocyte.

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Antimicrobial finishes for Textiles - Juniper Publishers

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Abstract

Infestations by micro-organism instigate cross disease by pathogens and odor develops in fabric have direct contact with the skin. Moreover, discoloration, tints and loss of functional characteristics of textiles are consequence of microbial damage. Antibacterial finished textile is an important area for medical and hygienic applications and there is enormous need of non-toxic and eco-friendly antimicrobial agents. The synthetic biocides finishes extensively reported were polyhexamethylenebiguanide (PHMB), quaternary ammonium compounds (QACs), metals (including metal oxides and salts), triclosan and n-halamines. Whereas, the natural based biocides (aromatic compounds, dyes, essential oil), chitosan and antimicrobial peptides (AMPs) were mainly considered among plant-based extracts. This paper will cover briefly, review of the latest research work on antimicrobial finishing, types of finish agents and various current developments in antimicrobial finishing to minimize the risks associated with application of organic, inorganic and plant based antimicrobial finishes

Keywords: Antimicrobial; Organic and inorganic finishes; Life; Textiles

Introduction

The major use of the antimicrobial was in the medical and the pharmaceutical industry. However, newer applications are possible. The textile fibers are these days increasingly treated with antimicrobial reagents. The other examples include the applications in food packaging and food storage, and medical, surgery and hygienic products etc. [1-3]. With the improvement of life standards, the demand of hygienic products is increasing for biocidal finishes in textiles (sports-wears, undergarment, bed-linen) and water filtration. The antibacterial finish treatment has become vital area of medical, surgical and healthcare activities due potential pathogenic microorganisms present in hospital environment and cause cross-infection diseases [4-8]. The types of micro-organisms include different kinds of organisms such as virus, bacteria, unicellular plants and animals, certain algae and fungi. Classification in bacteria family is “gram positive, gram negative, spore bearing or non-spore bearing type”. Some of the bacteria are of pathogenic nature that may cause infections to human [9]. A microbe (e.g. bacteria and fungus) normally protected with an outer cell wall that is composed of polysaccharides. The cell wall keeps up the veracity of cellular components and protects the cell from the extracellular situation; below the cell wall is a semi-permeable membrane that holds intracellular organelles, enzymes and nucleic acids. Chemical reactions within cell wall take place due to the enzymes present in cell wall. The nucleic acids hold the entire genetic directory of organisms [10]. The microorganisms responsible for microbial damage are generally present in surroundings; besides, formation of the substrates and the chemical processes may encourage growth of the microbes; further moist and warm environment still exaggerate the problem [11]. A gram-positive bacterium contains peptidoglycan and teichoic acid, peptidoglycan comprises of 90% of cell walls and made of amino acid and sugar. One example of gram-positive bacteria is Staphylococcus aureus that is in form of pair, short chain or graphic like cluster. Its size range is 0.5μm to 1.0μm and grows in temperature range of 35 to 40 °C.

Staphylococcus aureus is major cause of cross infection in hospital environment and 19% of total surgical infection. It’s also responsible for boils and also cause scaled skin infections. Other gram-positive bacteria are Staphylococcus epidermidis, Streptococcus pneumonia, Streptococcus pyogenes and Steptococcus viridians. The gram-negative bacteria are alike to gram positive bacteria apart from an outer layer of membrane affixed to peptidoglycan by lipoproteins which used to transport too low molecular weight substances. Gram negative bacteria are firm to diminish has compare to gram positive bacteria for the reason that of extra cell walls. An example of gram-negative bacteria is Escherichia coli (E. coli); its shape is similar to a bacillus and dwell in intestine of human. Escherichia coli can be proliferated during eating and/or usage of raw food stuff. The indications of E. coli are result in rigorous diarrhea (especially in kids) and kidney destruction. Other bacteria of this class are Klebsieella pneumonia, Pseudomonas aeruginosa, Salmonella typhi, Salmonella enteritidis and Haemophilusinfluenzae etc. [12]. Infestations by micro-organism instigate cross disease by pathogens and odor develops in fabrics that are worn after to skin or having direct contact with the body mass. Moreover, discoloration, tints and loss of functional characteristics of textiles are consequence of microbial damage [13-17]. Fungi moth or mildews are organism with lower progress ion rate; they stained the substrate and damage the fabric functional characteristics. Algae are classic microbes that either be fungi or bacteria, generate darker stains on the fabric surface [18]. Dust mites’ dwell in the home textiles and bed linen items include blankets, bed sheet, pillows and; especially in mattress and carpets. The dust mites feed on human skin and causes allergic reaction by healing waste products.

Antimicrobial Finishing Process

“The antimicrobial finishing process imparts the ability, to textile substrate, to inhibit the growth (-static) or reproduction of at least some types of microorganisms or to kill (-cidal) at least some types of microorganisms” [19,20]. Therefore, an antimicrobial finish should be capable to kill the microbes by breaching the cell wall or alter cell membrane permeability, obstructing the synthesis of proteins of microbes, blocking enzyme production necessary for microbes’ food. A few established antimicrobial agents, e.g. silver, quaternary ammonium compounds (QAC), N-Halamines; triclosan [21- 28] and polyhexamethylenebiguanide (PHMB) all are almost biocides [29-31].

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Finishing Mechanism

Three finishing mechanisms may be recognized based on the antimicrobial function performed by the particular finish on the textile. These mechanisms include control-release, regeneration and barrier-block. The first two finishing mechanism having problems in usage

The problems with control release mechanism are its durability after laundering and leaching of antimicrobial agent from fabric which can come in contact with wearer’s skin. These agents have the potential to affect the normal skin, which could lead to extreme skin irritation and allergy issues. These problems can occur with the fabric using a regenerate mechanism as these agents require chlorine bleach to activate its antimicrobial properties after laundering. Chlorine bleach not only damages the cotton fabric but is also harmful for human skin. Barrier- block mechanism does not pose the problems associated with other two methods. These agents are bonded on fabric surface and do not leach, thereby killing the bacteria that come in contact with the fabric [32].

Classification of Antimicrobial Finishing Process

The antimicrobial finish can be applied by physical and chemical methods, and by adding functional agents on to textile fibers. Such functional finishes can be of two main types, i.e. temporary antimicrobial finish and durable antimicrobial finish. The temporary finishes may lose easily while come into contact with skin or body fluids or during washing process because of weak bonding of finishing agent with fibers surface. Durable finish can generally be achieved by adding an antimicrobial finishing agent into fiber or textiles in wet processing, this method also known as controlled release mechanism. In such treatment, the finishing agent itself bonded with the fiber surface or a bonding substance may be used. The treated textiles deactivate bacteria by slowly releasing the biocide from the fiber or fabric surface [32].

Types of Antimicrobial Finish

A variety of chemical agents are available that may impart significant effect in textile fibers to inhibit the growth of microorganism. The important types of antimicrobial chemical agents are described in the following sections

Organic antimicrobial agents such as quaternary ammonium compounds (QACs),N-Halamines, Polyhexamethylene Biguanide; triclosan,; silicon based quaternary agent [33]; iodohors, phenols and thiophenols, heterocyclics, inorganic salts, nitro compound, urea, amines and formaldehyde derivatives, have been applied for antimicrobial treatment of textiles [34].QACs have been tested for antimicrobial activity of protein base wool, cellulose base cotton, synthetic base polyamides and polyester, the MIC value 10-100mg/l presented good reproducibility and good washing durability. These formations kill the microbes by altering cell membrane permeability, obstructing the synthesis of proteins of microbes, blocking enzyme production necessary for microbes’ food. The N-halamine compound is used for the development of antimicrobial cotton fabric through pad-drycure process followed by the exposure to chlorine bleach. The chlorinated sample showed potential antimicrobial ability against gram +ve and Gram –ve pathogens. It was experimented that on chlorinated after 15 days storage 85% of chlorine could be recharged that shows N-halamine compounds have good biocidal efficiency for healthcare textiles. Another organic based antimicrobial agent, Triclosan has been investigated for its antimicrobial ability for polyester, nylon, regenerated cellulose and acrylic fibers; with MIC value below than 10ppm versus bacteria against. Triclosan has excellent durability after use/washing and it prevents microbial growth by obstructing lipid biosynthesis. The most acceptable organic agent used for healthcare procedures, pharmaceutical and food industry is Poly-hexamethylene biguinide (PHMB). It’s efficient against both types of bacteria, in addition to yeasts and fungi. PHMB is slightly toxic and fewer skin infection issues were reported. It used in variety of products including undergarment and towel fabric to obstruct microbial growth and exhibited good washing durability. PHMB is bacterio-static at 1-10mg/l but at elevated values its bactericidal activity and inhibition rate raise collectively. The utmost antibacterial inhibition action of PHMB obtained between 5-6pH value [35,36].

Inorganic antimicrobial agents

The inorganic finishing agents such as metal oxides, copper and zinc, titanium, magnesium, silver and gold were applied for antimicrobial effects on textiles. These agents exhibited good durability for cellulose, protein, regenerated and synthetic materials with MIC value 0.05-0.1mg/lversus gram negative bacteria, E.coli. Sliver is wide acceptable inorganic antimicrobial agent and kills microorganisms by blocking and disengages the intracellular proteins. However, silver is a slight toxicagent, it releases slowly and can worn-out of the fabric [37-40].

Zeolites of chabazite-type with its optimal morphology and lowest silicon to aluminum ratio (Si/Al)solution that replaced with different combinations of silver, copper, and zinc ions to prepare single, binary, and ternary metal cation-modified zeolites were experimented and silver based zeolites exhibited more antimicrobial activity than the others and demonstrated good/suitable mechanical characters and excellent biocide effect against food borne bacteria and fungi on green polyethylene developed based on injection-molded composite. Further, the result confirmed its capability to rule the propagation of dangerous pathogens in environment of food processing and storage. Thus, these innovative antimicrobial materials are prospects for hygiene surfaces, kitchen accessories and packaging applications [41].

Limitations of inorganic and organic agents

In general, antibacterial property of any inorganic finishing agent is established with its chemical components. The biocide efficiency of inorganic agents slowly drops in use and during wash. The most of such agents carry limited intensity of microbes’ inhibition, moreover they are poisonous, initiate skin problem to humans and having problem to decompose in down streaming [42,43]. To reduce the risks allied with the application such inorganic agents, there is enormous need of substitute agents for antimicrobial treatment of textiles. As mentioned early, a wide range of organic antimicrobial agents are available for textiles treatment but out of these agents; triclosan, quaternary ammonium compounds, Polyhexamethylene Biguanide have been used on commercial scale. Polyhexamethylene Biguanide is slightly contaminated with poisonous concerns and hard to decompose in down streaming. In US Preregistration Eligibility Decision for PHMB by US Environment Protection Agency” the discharge of effluents containing PHMB is not allowed without mandatory treatment.

Eco-Friendly Antimicrobial Agents (Natural Plant and Fruit Extracts)

Plant extracts provided an attractive source of eco- friendly antimicrobial finish. The natural cure using plant extracts is increasingly receiving interest in the development of antimicrobial textiles. One of the plant-based sources is belong to Meliaceae family Neem (Azadirachtaindica); it is one of the most prominent from natural gifted sources of antimicrobial compound. All parts of neem are established for potential antimicrobial constituents. The extract from each part of the neem presented active antimicrobial effectiveness to block the proliferation of the bacteria. Currently, a small number of studies has demonstrated neem’ used for textiles to evaluate it antimicrobial activity. However, cotton and cotton/polyester blended fabric treatment with seed and bark extracts were reported [44-46]. Moreover, the cotton fabric imparted with neem leaf extract loaded nanoparticle [47]and synthesis of sliver nano-particles using extract of neem leaf for cotton treatment was also used [48]. Another plant-based source belongs to the Liliaceae family Aloevera (Aloe barbadensis), its leaf extract has antibacterial and antifungal potential and have been used for dressing gauzes, sutures and other medical textile applications [49,50]. Similar to neem applications in textiles, a few studies of aleovera application for cotton fabric treatment were articulated. However, more research is required. Antimicrobial finishing of cotton and cellulose fibers is significantly useful and important in medical textiles utilization [51]. One of the other plant-based sources is Ginkgo biloba or Ginkgoceae (Mantissa Plantarum Altera). Ginkgo biloba tree has flourished in jungles for more than 150-250 million years. It is assumed to be one of the aged living species on earth [52-54]. The standardized extract formulation of ginkgo leaf in used hold “5-7% ginkgolides and bilobalide (BB) [55]”. It is an excellent candidate for antimicrobial treatment of healthcare cotton textiles. The formulation of Ginkgo biloba extract standard values were forced because cyto-toxicity issues was reported beyond these limits [56-58]. Jang and Lee investigated ginkgo leaf extract antimicrobial activity for Tencel fabric in extract formulation containing silicon softer along with crosslinking agent.

The study concluded that Ginkgo Biloba extract is ecofriendly antimicrobial agent and their application was investigated in health and medicinal purposes. It is exclusively for non-toxicity characteristics linked with such other agents, it is potential candidate for antimicrobial finishing of institutional textiles range including home accessories and hospital bed sheet, nurses’ uniforms, surgical gown and drapes etc. [59]. The plant based natural fruit source reported for antimicrobial properties, the fruit-based source is Pineapple (Ananascomosus) juice was investigated against harmful microbes60. The antimicrobial activity was evaluated through agar diffusion method. Another plant base source reported is Papaya (Carica papaya). Its fleshy tissues hold three influential antioxidants i.e., vitamin A, C and E. Further, it contains stuff of proteolytic enzymes that have good antimicrobial activity against bacteria, fungi and virus. The papaya fruit seeds are spicy and very strong that yield them almost indigestible. These seeds have more potential pharmaceutical worth as compare to it’s the flesh and are effective against bacterial infection. The juice presented the excellent antimicrobial ability versus a number of gram native bacteria [60-63]. Moreover, the uses of both these fruit juices/ extracts were not reported to assess their biocide or bio-static activity.

The medicinal plants Clove (Eugenia caryophyllata), Falsedaisy (Eclipta alba), Leadwort (Plumbagozeylanica), and Mint (Mentha Arvenesis) parts were dried, powdered, grinded and extracted with solvents and applied through pad-drycure and microencapsulation techniques. The fabric samples were then subjected to antimicrobial testing and the bacterial growth was analyzed after 5, 10, 15 and 20 washing cycles. The antimicrobial activity of microencapsulated finish was effective till 15 wash cycles64. Several other plant base dyes are reported for their antimicrobial and antifungal activity such as Henna (Lawsoniainermis), Walnut and alkanet (Anchusa Tinctoria), curcumin, pomegranate, cutch, red onion peel and a mixture of red onion peel/curcumin (40 g/L, 50%)65. The extract of neem (Azadirachataindica), Lam (Buteamonosperma) and Gaetin (Litchi chinensis) trees was used to check antibacterial, antifungal activity and aesthetic properties (stiffness and appearance) of 100% silk fabric. It was confirmed that the formulation of antimicrobial finish improves the aesthetic properties. It is further reported that treated finished showed good/suitable/optimize results and 89% reduction in microbial growth was achieved up to 25 washes66. Application of the plant base dye-stuff is the art of imparting hues and tints to textile substrate. Dye-stuff or coloring matters acquired from natural resource are tested for antibacterial activity of the fabrics and results of the dyed fabrics presented these days have effective antibacterial activity. Although, synthetic dyes contain a range of vibrant color and are extensively used but now a day, natural dyes gaining interest because of strict environmental standard forced by a number of European states due to carcinogenicity and photosynthetic issues of synthetic dyes. Natural dyes are considered as eco-friendly, nontoxic, medicinal features [67]. The essential oil extracted from Rosemary (Rosmarinusofficinalis) and orange (Citrus sinensis) were obtained by steam distillation from rosemary vegetal mater and orange peel used to evaluate the antimicrobial activity for textile substrate (56% cotton/44% polyester) with concentration of 1%, 3% and 5%of each oil and antimicrobial activity was assessed against each strain. The demonstrated results support textiles functionalized with rosemary and orange essential oils, both are efficient active antimicrobial barriers with maximum reduction of 56.99% for rosemary and 92.48% reduction for orange essential oil [68].

Plant based Bamboo material is well known for their antimicrobial ability. In presented reported study focused to evaluate the antimicrobial property of plasma treated bamboo fabric imparted with combinatorial herbal extract. The knitted bamboo fabrics were rendered to plasma treatment at most appropriate setting to improve the hydrophilicity. The variations in the hydrophilic characteristics, physical and chemical changes of the plasma treated fabric were measured by using standard tests and combinatorial herbal powder was subjected to different solventextracts and their antimicrobial efficiency against pathogens were evaluated. The ethanol herbal extract presented higher antimicrobial activity against E. coli (12mm), S. aureus (14mm) for zone of inhibition and tests proved wash durability retained till 25 washes [69,70].

The chitosan and alginate have been used for antimicrobial finishing of textiles. Chitosan is derivate of chitin, water-soluble cellulose based. Chitin is a polysaccharide base on amino sugars. In an acid solvent amine component turn into quarterly amino unit that inhibits growth of microbes. Theses amino unit performs as shield to block protein and slow down proliferation by distracting cell membrane; this permit the substance to escape from bacterial cell, consequential results is death of the bacteria. Antimicrobial activity of chitosan was reported in many studies and it is widely accepted antimicrobial agent [71]. The β-Cyclodextrin, Chitosan citrate and β-Cyclodextrin/Grafted Chitosan with lavender essential oil were also used to evaluate the combined effect of fragrance and antimicrobial activity on cotton textiles through pad-dry method. The results discovered that β-CD was highly soluble in 0.6g/l NaOH solution and 80 gpl β-CD and 6% essential lavender oil solutions were found to be a most suitable combination for fragrance and antimicrobial property [72]. In another study, the most common polymers polypyrrole (PPy) was used with its environmental stability, ease of synthesis, exciting chemical, electrical, electrochemical and optical properties [73]. The antimicrobial activity of polypyrrole-graftchitosan copolymer was investigated by chemically synthesized, and then its composition and morphological characteristics were evaluated. The results discovered the strong interactions among polypyrrole and chitosan chains. Further, the electrical conductivity of chitosan increased to semi-conducting level by grafting. The thermal stability and crystallinity of polypyrrolegraft- chitosan copolymer increased while compared to chitosan. The copolymer was evaluated versus various bacterial and fungal strains at different concentrations and results achieved were evaluated with the reference antibiotics and it was pointed that the polypyrrole-graft-chitosan copolymer has stronger antibacterial activity than the polypyrrole and the chitosan alone; and it further increased at higher concentrations [74]. Monica Periolatto reported, the sound fastness and stability was attained with both photo-grafted chitosan and polypyrrole coating on textiles. It was proposed that a synergic impression of polypyrrole-chitosan finish, exploitable in textiles [75].

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Challenges Associated with Plant Finishes

The chitosan is one of the recognized bioactive agents used on commercial scale for fabric antimicrobial finishing. On the other hand, its effectiveness is spoiled by a few factors. It depends on the chitosan molecular weight, pH value, ions intensity, add-on of non-aqueous solvents and the grade of deacetylation. Moreover, their treatments for textiles are efficient at maximum concentrations consequently reduce the air permeability of fabric and impart stiffness. Normally, herbal extracts including chitosan evaluated for their antimicrobial activity for textiles reported various issues, such as problem in extraction, separation of bioactive substances, textiles treatment with bioactive agents and the most important concern is poor finish durability after uses and during washing. Regardless of few most important challenges linked with plant based antimicrobial finishes, nevertheless these extract formulations are appealing with their non-toxic and environment friendly characteristics [76,77].

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Role of Nanotechnology in Antimicrobial Finishing

Nanotechnology may provide finishes to combat infectious pathogens. Their application through nanotechnology engaged several parameters that control, manipulate and assemble nanoscale constituents to develop materials, systems or devices. Studies reported that the silver nano-particles exhibit excellent antimicrobial property versus microorganisms. For examples, the expedient use of antimicrobial metals such as zinc, copper and silver were incorporated into an FDA-approved polymer (polycaprolactone– PCL) to produce filaments. Hot melt extrusion was used to extrude pellets obtained by vacuum-drying of solutions of PCL and the different metals in order to manufacture metal-homogeneously-loaded filaments. Wound dressings with different shapes were produced with the filaments containing different concentrations of metals. The antibacterial efficacy of the wound dressings was tested using a thermal activity monitor system, revealing that silver and copper wound dressings had the most potent bactericidal properties [78].

Now a day, metal oxide nanoparticles (MeO-NPs) become a potential substitution to combat toxic infectious complaints and substantially resistant to different types of antibiotics [79]. ZnO particles nano-structured use on the cotton textile surface with different surfactants to stabilize, homogenize the coating and has improved the durability of ZnO NPs with decreased its leaching and showed the highest antibacterial and antifungal activities against different pathogenic bacterial and fungal species with high reduction reached over 90% [80]. Another technique used to investigate the antimicrobial property, i.e. application of Znonano-particle and use soluble starch as capping agents revealed that antimicrobial activity is oversee by the type of capping agents and results in achieved lower particle size of 3-5nm and higher antimicrobial rate as compared to other capping agents [81].

The use of Copper nano particle/nano-composite for antimicrobial ability in glycerol-polyvinyl alcohol matrix in gel and moldable plastic form proves that it can be produced and easily figured at high temperature. The materials show very good long-term stability in air, protecting the produced copper nano-particles from oxidation and proven inhibition of bacterial proliferation of both Escherichia coli and Enterococcus faecalis bacteria in nano-composite existence [82]. Biocompatible nanogold (AuNPs) have gained considerable attention for potential applications in nano-medicine due to their characteristic size dependent chemical, electronic and optical properties and displayed antibacterial efficacy towards different bacterial species and the MIC was evaluated to be 960μL/ml against S. aureus [83].

To overcome the toxicity and washing durability problems associated with plant-based extracts, application of chitosanneem nano composites for development of antimicrobial cotton was used. Silver nanoparticles micro-gel based on poly-(N-isopropylacrylamide) and chitosan [84]; and chitosan nanoparticles loaded with Fe2+ or Fe3+ surfactant-assisted chitosan chelating Fe2+, Fe3+ and ionic gelation chitosan showed very high antimicrobial property at lower concentrations as compared to chitosan [85]. In another study, the results confirmed the biosynthesized AgNPsusing pre-hydrolysis liquor of Eucalyptus wood as effective growth inhibitors against microbes for various biomedical applications [86]. Further, Chitosan and acrylic acid bi-grafted polypropylene melt-blown nonwoven membrane immobilized with silver nanoparticles presented excellent antibacterial and hydrophilic properties [87].

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Conclusion

An interesting variety of antimicrobial finishing agents is available. However, limitations are possible to provide acceptable performance, environment-friendly traits, and cost requirements. Majority of inorganic antimicrobial agents are poisonous, potential problem to degrade in environment, inhibited a limited range of microbes and possess poor laundering durability; but comparatively organic agents have lower adverse effects. The use of nano-particle has improved the efficiency of some of the present use antimicrobial agents and reduced the environmental issues associated with these agents (such as toxicity and washing durability) and exhibit excellent antimicrobial property versus microorganisms. Moreover, despite the washing durability challenge associated with natural plants based antimicrobial finishes; they are widely accepted antimicrobial agents for textiles finishing with their eco-friendly and non-toxic characteristics. Use of plant based nano-particle antimicrobial agents has been growing in many different fields primarily due to their advanced characteristics and protection against pathogens as comparison to conventionally used biocides and such value-added finishes may provide sustainable healthcare applications in textiles

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Juniper Publishers| Overview and Future of Hemo-Components and Natural Guided Regeneration

Journal of Surgery-JuniperPublishers

The History of Platelet Rich Fibrin (hemocomponents) started in 1970, when Matras described a fibrin glue, formed by polymerizing fibrinogen with thrombin and calcium, which was used to improve skin wound healing in a rat model in 1970 [1]. Because of the low concentration of fibrinogen in plasma, the stability and quality of fibrin glue were low. A few years later several research works proposed an upgraded concept for the use of blood extracts, termed “platelet-fibrinogen-thrombin mixtures” or “gelatin platelet - gel foam” [2,3]. In this new concept, the fibrin glues were presenting a significant concentration of platelets within the final preparation. The idea was first to reinforce naturally the fibrin gel, and also to combine the healing properties of the platelets with those of the fibrin. This improvement allowed to prepare more natural products, integrating more natural blood constituents as it should.These products were the first platelet-rich plasma gels. These new strategies insisted in the role of platelets within the fibrin gel, and offered excellent preliminary results in ophthalmology, neurosurgery and general surgery. Whitman proceeded to develop this technique in 1997 and particularly Marx et al. [4,5] in 1998. The Leukocyte- and Platelet-Rich Fibrin L-PRF clot was often described as “optimized blood clot” that can be surgically handled and used. The rationale to use this glue/membrane and its success is due to fibrin, platelets, growth factors slow release, leukocytes and other cells: all these components are the key active actors of the natural healing process and combined together are forming a kind of engineered tissue extracted from the blood circulating tissue [6].Unfortunately at the moment there is a lack of an international standard for characterization, classification and identification of surfaces in implantable materials [7,8], in particular a standardization is needed to obtain an optimal and reproducible results, however the current classification of platelet-rich concentrates is based on their fibrin architecture and cell content. It consists in two main groups of products, platelet-rich plasma (PRP) and platelet-rich Fibrin (PRF), both of which are available in a pure or leukocyte-enriched form (L-PRP and L-PRF) [9]. Each product has an unique biological profile that dictates its clinical applications. L-PRF concentrates provide slow release of many growth factors and can be easily prepared during surgery [10-14]. They are inexpensive and autologous; therefore, they avoid the complications associated with allogenic blood use.Pure Platelet-Rich Plasma (P-PRP) products are preparations without leukocytes and with a low density fibrin network after activation. One largely advertised method of P-PRP is known under the commercial name PRGF [Plasma Rich in Growth Factors or Preparations Rich in Growth Factors or EndoRet, Biotechnology Institute BTI (dental implant company), Vitoria, Spain] and was tested in many clinical situations, particularly in sports medicine. P-PRP gel released most of its growth factors in the first hours and completely dissolved in the medium after 3 days, even after a maximum artificial fibrin polymerization.Leukocyte-and Platelet-Rich Plasma (L-PRP) products are preparations with leukocytes and with a low-density fibrin network after activation. The methods to prepare the PRP membranes require two or one centrifugations, there are, infact, some new faster machines like Arthrex ACP®, nevertheless an anticoagulant is always needed. PRP families are not adapted (complicated, expensive, with mixed clinical relevance) for daily oral applications. PRP families are substitutions to fibrin glues in most other surgeries, particularly to improve skin wound healing. The use of gelling of the PRP on the surgical site makes it adequate surgical adjuvants in many clinical situations, even if the exact effects - in comparison to fibrin glues - remain largely debated.The PRP solutions have also the advantage to be liquid before activation, and can therefore be used as injection or placed during gelling on a skin wound or suture (similar to the use of fibrin glues) in various sports medicine or orthopedic applications. In this strategy of regenerative medicine, the platelet suspensions are injected like other pharmaceutical preparations. The results of this method remain however largely debated in the literature, probably because of the large quantity of different protocols [14-16]. Pure Platelet-Rich Fibrin (P-PRF) - or Leukocyte- Poor Platelet- Rich Fibrin preparations without leukocytes and with a high-density fibrin network. These products only exist in a strongly activated gel form, and cannot be injected or used like traditional fibrin glues. However, because of their strong fibrin matrix, they can be handled like a real solid material for other applications.L-PRF membrane remains solid and intact after 7 days and relases continuously a large quantity of growth factors, a significant part of it being produced by the cell population within the membrane. L-PRF family fits the needs of the applications in oral and maxillofacial surgery, as L-PRF clots and membranes present a volume and shape easy to combine with most surgical techniques, as filling and interposition healing biomaterial or as protection healing membrane. The fibrin architecture of L-PRF is constitued by connected trimolecular junctions, due to a slow polymerization of the platelet concentrate and due to the absence of heterologous thrombin. The results of this process is a flexible fibrin network, able to promote the gradual release of growth factors and leukocytes migrationduring extended period.It is easy to prepare in large quantity and inexpensive, what makes it particularly adapted for daily clinical practice. PRF families in general are usable in other disciplines with interesting results, particularly for the treatment of skin chronic wounds and ulcers. The methods to prepare PRF never require an anticoagulant and a lower G-force is needed (around 400G). PRF products cannot be used as injectable products in sports medicine for example [12,17]. Some groups advocated that the presence of leukocytes may be negative for the therapeutic outcome, due to a potential risk of stimulation of the inflammatory process after the membrane placement in a wounded site [18]. Other researchers insisted on the need of some leukocyte population in the injectable PRP in order to increase the growth factors production, the release of anti-pain mediators and the natural anti-infectious activity.Some kind of leukocytes, lymphocytes in particular, are playing a key function as regulation turntable of the healing and inflammatory process, and there is no reason to discard them. Leukocytes are not only inflammatory cells: they also present anti-nociceptive effects through different chemokines, anti-inflammatory cytokines (IL-4, IL-10 and IL-13) and opioid peptides (b-endorphin, metenkephalin, and dynorphin-A) and can therefore promote a clinically relevant inhibition of pathological pain [19-21]. The classification previously described is the only nomenclature which considers all forms of platelet concentrates for surgical use. However, other classifications systems were proposed in the recent years, but are limited because they only refer to Platelet-Rich Plasma products and sports medicine applications. Both proposals are not significantly evidence-based and do not allow to improve the current terminology [22].Most publications about growth factors and platelet concentrations showed the relative lack of significance of these parameters, due to the many inter-individual variations and the short-term effects of these parameters: platelets being activated and active during a very short time and the growth factors being released, consumed locally or dissolved in the blood circulation in few minutes or hours after their release [23,24]. Platelet concentrates for surgical use are a system of all blood elements within a logical healing platform including the fibrin matrix, the platelets, the mediators and the cells all together to reach a clear and reproducible clinical result [25]. Castro in a systematic review founded favorable effects on hard and soft tissue healing and postoperative discomfort reduction were often reported when L-PRF was used, nevertheless, they found a lack of standardization of the protocol in regenerative procedure [26].Temmerman et al. [27] compared bone ridge preservation L-PRF socket filling and natural healing following tooth extraction after 3 months; the results showed the use of L-PRF as a socket filling material in order to achieve ridge preservation is beneficial for all parameters considered (vertical height changes, width reduction, mineralized bone) during a 3 month observation period. Furthermore, the use of L-PRF results in less post-operative discomfort and pain for the patients. Multiple surgical specialties have recognized the potential advanatges of platelet-rich concentrates. Their use has been described in ophthalmology, neurosurgery, general surgery [22] orthopedic surgery, sports medicine [28] and oral and maxillofacial surgery [29]. Several applications of L-PRF concentrate have been described in the literature including postoperative hand wound healing yielding faster re-epithelialization and in the treatment of androgenic alopecia diminishing hair loss among others [30- 32].The role of L-PRF in endoscopic endonasal skull base surgery defect reconstruction was investigated by Soldatova et al. [33] who demonstrated the potential benefits of L-PRF membranes for the reconstruction of skull base defects with encouraging rate of healing progression as measured by the crusting score. During the lasts years the production of platelet concentrates for surgical use from the PRF (Platelet-Rich Fibrin) family are becoming very popular in some surgical fields. The main product is classified as L-PRF and is used in oral and maxillofacial applications in particular. Many systems are available on the global market, but only one system to date is duly CE-marked and FDA-cleared (Intra-Spin System, Intra-Lock, Boca-Raton, FL, USA) [34].The impact of the centrifuge characteristics and centrifugation protocols on the cells, growth factors and fibrin architecture of L-PRF was investigated comparing 4 different centrifuges. The results showed significant differences in the vibrations level at each rotational speed between the 4 tested machines. The CE-marked and FDA-cleared device was the most stable machine in all configurations and it remains under the threshold of resonance, unlike the 3 other tested machines [35]. In another study M.F-Kobayashi demonstrated in vitro that reducing the centrifugation speed favored an increase in growth factor release from PRF clots which in turn may directly influence tissue regeneration by increasing fibroblast migration, proliferation and collagen mRNA levels [36].Go to

Conclusion

L-PRF treatment offers additional advantages: favorable effects on hard and soft tissue healing, postoperative discomfort reduction, simple harvesting, simplicity in use, no need for primary closure, and no risk for early membrane exposure. The economic implication in the final cost of a treatment has also to be taken into consideration. The vitro and molecular biology studies are very useful to understand which molecules are present in the clot and to hypothesize their role in the healing and regenerative process, however more clinical standardized studies are needed to demonstrate the quantity of growth factor is actually necessary to significantly improve the regenerative processes. Literature’s results are often discordant, several practitioners report different clinical experiences and mixed clinical outcomes. These unpleasant facts are due to a chaotic market and a lack of standardization of the procedure. Further researches and clinical trial under a rigid protocol are needed to fully understand the potential and optimal effect of L-PRF in regenerative procedures. To read more articles in Journal of Surgery Please Click on: https://juniperpublishers.com/oajs/index.php For More Open Access Journals in Juniper Publishers Click on: https://juniperpublishers.com/journals.php

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Juniper Publishers-Open Access Journal of Case Studies

Clinical, Radiographic, and Histologic Outcomes of Ankle Arthrodesis in a Diabetic Patient using a Biphasic Calcium Phosphate Bone Graft with a Novel Submicron Needle-shaped Surface Topography

Authored by Thomas Fusco

Abstract

Patients with diabetes mellitus who sustain ankle fractures are at increased risk of complications including non-union, malunion, wound dehiscence, soft tissue infection, osteomyelitis, deterioration to Charcot arthropathy, and amputation [1-3]. Multiple publications have evaluated treatment options for diabetic patients with ankle fractures; management options include non-surgical treatment and immobilization, open reduction internal fixation, arthrodesis of the tibiotalar joint, arthrodesis of the tibiotalocalcaneal joints, external fixation, and amputation [4-7]. It is important to identify an effective bone graft for these cases, as well as to understand the science behind the bone graft selected. In recent years, the study of osteoimmunology has confirmed the reciprocal interaction of bone forming mesenchymal stem cells and immune cells on bone growth. This report describes an adult female patient with a late presenting ankle fracture dislocation, initially treated with ankle arthrodesis using a novel bone graft with submicron needle-shaped surface topography. The patient developed Charcot arthropathy and was revised to a tibiotalocalcaneal (TTC) nail. During revision, a biopsy was taken of a solidly fused segment and sent for histology. Radiographic, clinical, and histological data were tracked and showed solid fusion at 6 weeks radiographically, which was confirmed via histology and surgical exploration at 12 weeks, indicating notable results in this challenging arthrodesis case using a novel bone graft with submicron needle-shaped surface topography.

Keywords: Charcot arthropathy; Diabetic ankle fracture; Diabetic ankle arthrodesis; Osteoimmunology, Mesenchymal stem cells; Surface topograph

Introduction

Patients with diabetes mellitus who sustain ankle fractures are at increased risk of complications including non-union, malunion, wound dehiscence, soft tissue infection, osteomyelitis, deterioration to Charcot arthropathy, and amputation [1-3].There is no gold standard for treatment, as the success of both bony and soft tissue healing depends on factors such as fracture pattern, patient co-morbidities, hemoglobin A1c, the presence of neuropathy, the ability of the patient to remain non-weight bearing, and others [3]. Multiple publications have evaluated treatment options for diabetic patients with ankle fractures; management options include non-surgical treatment and immobilization, open reduction internal fixation, arthrodesis of the tibiotalar joint, arthrodesis of the tibiotalocalcaneal joints, external fixation, or amputation [4-7]. In 2021, a meta-analysis of open ankle arthrodesis reported an overall complication rate of 14% for this procedure [8]. In diabetic ankle fractures the reported complication rate is much higher, with one review reporting a complication rate of 47% [9]. One important challenge of arthrodesis in diabetic patients is identifying the most scientifically advanced bone graft for the procedure [10]. Synthetic calcium phosphate bone grafts closely resemble human cancellous bone, have an exceptional safety profile, and have proven to be a cost-effective alternative to autograft [11-13].

In response to tissue trauma or surgery, the immune system reacts by upregulating pro-inflammatory macrophages, also known as the macrophage M1 phenotype. If M1 macrophages are chronically activated a fibrotic healing response is initiated, ultimately leading to a non-union. Conversely, if the macrophage M2 phenotype is activated a pro-healing response is initiated, upregulating mesenchymal stem cells and promoting the formation of bone rather than scar tissue [14]. Utilizing a biphasic calcium phosphate (BCP) bone graft with needle-shaped submicron surface features (BCP<µm) polarizes naïve monocytes to the pro-healing M2 phenotype. In preclinical studies, this BCP<µm has been shown to promote bone formation even in soft tissue, without the need for added cells or growth factors [15]. This novel BCP<µm bone graft mimics the structure of cancellous bone, with bone formation taking place throughout the bone graft leading to a solid, uniform fusion that has been demonstrated in clinically relevant animal models [16,17].

This case describes an adult female patient with a late presenting ankle fracture subluxation, initially treated with ankle arthrodesis using a novel bone graft with submicron surface topography. The patient developed Charcot arthropathy and was revised to a tibiotalocalcaneal (TTC) nail. During revision, a biopsy was taken of a solidly fused segment and sent for histology. Radiographic, clinical, and histological data were followed for 24 weeks from initial surgery and 12 weeks from revision surgery.

Case

The patient is a 73-year-old female who presented to the emergency department three weeks status-post left ankle injury. The patient reported that three weeks prior she sustained a left ankle injury and went to the emergency department where she was diagnosed with an ankle fracture dislocation and underwent closed reduction. She was discharged for outpatient follow up at that time. Upon outpatient follow up, the patient reported that radiographs were obtained showing loss of reduction and repeat dislocation. She stated that she returned to the emergency department where she was re-reduced and splinted. Again, the reduction did not hold, and she returned to the emergency department. At this time, the author of this paper became involved. Radiographs from the emergency department on that day revealed a trimalleolar ankle fracture with subluxation See Radiograph Series 1.The patient’s past medical history included diabetes mellitus type II with neuropathy, peripheral artery disease, hypertension, hypothyroidism, hyperlipidemia, coronary artery disease, gastroesophageal reflux disease, and depression. On admission, her hemoglobin A1c was 9.1, and her body mass index was 36.9kg/m2. She denied the use of tobacco. For graphic representation of the surgical and post-operative timeline, see Figure 1.

The patient was immediately admitted from the emergency department and scheduled for closed reduction and external fixator placement in the operating room. This procedure was performed uneventfully the following day. The patient was discharged home, planning to return to the operating room one week later for definitive fixation. At the time of definitive fixation, the external fixator was removed, and a tibiotalar arthrodesis was performed using a four-screw construct. This was fortified with placement of a new external fixator. During this procedure, a novel BCP bone graft with a needle-shaped submicron surface (BCP<µm)was used to pack the arthrodesis site. This surgery was uneventful, and the patient was discharged home as expected with instructions to remain non-weight bearing on the left lower extremity. See Radiograph Series 2.

Four weeks after definitive surgery, the patient presented to her post-operative appointment with concern for an infected external fixator pin. The decision was made to bring the patient back to the operating room for external fixation removal and thorough debridement. At that time the pin sites were cleaned and debrided, and questionable bone was explored with cultures sent to pathology. Clinically, the bone did not appear osteomyelitic, which was confirmed with multiple negative bone cultures. The patient was prescribed a full course of oral antibiotics.

Two weeks after removal of the infected external fixator, the patient returned for her six-week post-operative appointment. At that time, she was doing well. Her wounds were clean, dry, and intact, her edema was improving, and her radiographs showed a healing fusion site with hardware in good position without loosening or lucency around the screws Radiograph Series 3.

Three months status-post her definitive surgery, the patient returned for follow-up. Radiographs were obtained, which showed interval deterioration of her hindfoot with Charcot arthropathy collapse See Radiograph Series 4.A thorough discussion was held with the patient, and ultimately the decision was made to revise the tibiotalar arthrodesis to a TTC nail. During this procedure, the arthrodesis site was explored, and the novel BCP bone graft had integrated well into the fusion site with solid appearing bone and no migration of the granules. Biopsy of the fused segment was obtained and sent for histology See Intra-Operative Image.

Following the TTC nail, the patient had an uncomplicated post-operative course. At her 12-week post-operative office visit she had no complaints, and her radiographs showed a well-aligned healing arthrodesis. The patient was cleared to begin weight bearing. Shortly thereafter, the patient moved to another state and was lost to follow up See Radiograph Series 5.

Biopsy Results

The biopsy obtained from the revision surgery at the tibiotalar arthrodesis site twelve weeks post-operatively was processed for routine bone tissue histology with hematoxylin-eosin and Safranin O stains. The biopsy showed mature bone tissue with integration of the BCP<µm bone graft. Bone was observed in direct apposition with the BCP<µm particles, growing around and between the material and inside its pores. Bone tissue around the material had a normal phenotype, including woven and lamellar structure, the presence of blood vessels, and cells such as osteoblasts, osteoclasts, and osteocytes residing in lacunae. Ongoing new bone formation, as characterized by presence of osteoid matrix, was commonly observed directly against the surface of the BCP<µm particles or inside their pores. No adverse reactions to the BCP<µm bone graft were observed See Histology.

Discussion

Ankle arthrodesis in Charcot arthropathy is known to lead to increased complications such as non-union, malunion, infection, and amputation [2]. Successful fusion depends on a variety of factors including patient age and co-morbidities, arthrodesis technique and implants chosen, and bone graft selected [3].This case exhibits the benefits associated with the use of a biphasic calcium phosphate bone graft with submicron needle-shaped surface topography in achieving a reliable and solid fusion.

The patient, an elderly female with uncontrolled type II diabetes mellitus and Charcot arthropathy, is a difficult clinical scenario with a successful fusion outcome using BCP<µm bone graft. The patient had radiographic fusion via X-Ray at six weeks post-operatively from the index procedure. Additionally, due to the unforeseen complication of Charcot arthropathy at the joint below the fusion site, the surgeon was able to obtain histology from the original fusion site, which showed a significant amount of mature lamellar bone at only twelve weeks after the initial fusion with BCP<µm. The literature on specific bone grafts in ankle and hindfoot arthrodesis is limited; however, one large, prospective, randomized multi-center study evaluating rhPDGF-BB/β-TCP bone graft reported fusion outcomes of 61.2% as determined by Computed Tomography (CT) at six months post-operatively [18]. In contrast, this case was able to report fusion confirmed by X-Ray six weeks post-operatively, as well as by second look surgery and histology at twelve weeks post-operatively.

The case also highlights a successful fusion outcome in a procedure known to be technically difficult and fraught with risks such as infection and non-union. Recent literature reports high complication rates in both diabetic ankle fractures and TTC nails, with one review of diabetic ankle fractures reporting an overall complication rate of 47%, and another retrospective review reporting a non-union rate of 22.8% after subtalar arthrodesis via TTC nail [9,19].

In this case, the fusion was confirmed via multiple methods: clinically, radiologically, histologically, and via the gold standard for fusion assessment: surgical exploration [20]. Remarkably, this case demonstrated fusion via X-Ray at six weeks post-operatively, as well as mature lamellar bone formation via surgical exploration and histology at twelve weeks post-operatively. This report demonstrates that BCP<µm bone graft with needle-shaped submicron surface topography is an effective bone graft for use in complicated extremity arthrodesis procedures.

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Feasibilities of the Pinhole Surgical Technique: Mini Review- Juniper Publishers

       Juniper Publishers- Open Access Journal of Dentistry & Oral Health

Feasibilities of the Pinhole Surgical Technique: Mini Review- Juniper Publishers

Authored by Saad Al-Almaie

Abstract

The concept and technique of the pinhole surgical procedures has been a debated topic in dentistry in recent years. This paper consists of a mini review of the literature available on pinhole surgical technique. This review shows that pinhole surgical technique allows us to reposition the gums quickly and easily, with the less invasive method, a decreased patient discomfort, a shorter treatment and recovery time and much less pain.

Keywords: Pinhole surgical technique; Minimally invasive surgery; Recession

Introduction

Gum recession refers to the loss of gum tissue along the gum line. This can occur as a result of periodontal disease (gingivitis, periodontitis, advanced periodontitis), the natural aging process, or abrasive habits when it comes to brushing the teeth. When gum recession occurs, the root structure of the tooth becomes exposed and loses of the tissues covering the root. This means that tooth decay and other problems can affect the teeth along the gum line and beneath it. Since healthy gums are essential for a healthy mouth, getting gum recession treated is important for lasting dental wellness [1-3]. The surgeons are now interested in more user-friendly, less invasive, esthetically favorable patient oriented surgeries as they now aim at/prioritize patient comfort, satisfaction, and patient centered outcomes. The concept of “extension for prevention” has now changed to “conserve to preserve.” The concept of “minimally invasive surgery (MIS)” is one such peculiar and innovative approach which aims to produce minimal wounds, minimal flap reflection, and gentle handling of the soft and hard tissues [4]. Wickham and Filtz described the techniques of using smaller incisions as “MIS” which were later defined by medical subject headings as those procedures that avoid the use of open invasive surgery in favor of closed or local surgery. Hunter and Sackier described the same as “the ability to miniaturize our eyes and extend our hands to perform microscopic and macroscopic operations in places that could previously be reached only by large incisions [5].” In recent years a novel surgical approach to root coverage, called the pinhole surgical technique, has been gaining exposure. Chao introduced it for Miller class I, II, and III recession defects and reported favorable predictability for root coverage and defect reduction up to 18 months following the procedure [6]. Miller introduced his classification system [7]. The author related the extent of the soft tissue recession to the location of the mucogingival junction as well as the height of interproximal clinical attachment adjacent to the surface affected by the recession.

Review of Literature

Pub Med databases were used to search for published articles about pinhole surgical technique. Clinical studies and cases using this technique were included. Letters to the Editor, historical reviews, and unpublished articles were not sought. Some articles were directly excluded after reading only their titles. At this stage there were 9 to 10 articles included, and the inclusion and exclusion criteria were defined. Thirteen articles were included for full-text reading. After reading these, three more articles were excluded, as they did not fulfill the inclusion criteria. These articles were included in introduction and discussion in addition to that, more reviews of literature had been included regarding pinhole surgical technique.

Main Outcomes of Selected Studies

This treatment offers a more conservative alternative to common periodontal treatments, such as gum grafting. With the Pinhole Surgical Technique, the clinician makes a small hole in the gums, the size of a needlepoint, and then manipulates the tissue to correct recession and other issues. In addition, for some cases, a biocompatible collagen agent is inserted, filling the space between gingival tissue and the roots, which helps heal the area and allow healthy tissue to attach to the teeth. Most of the selected studies considered different benefits, including cosmetic improvement, arresting further breakdown, no need for scalpels or sutures, preservation of natural gum tissue, and reduced risk of periodontal problems [6,8-13].

Discussion

Zucchelli and Sanctis conducted a case series study to evaluate root coverage with a new surgical approach to the coronally advanced flap procedure for treatment of multiple recession defects in patients with esthetic demands at 1 year examination. The results showed 88% complete root coverage and greater reductions in recession in cases with less amount of keratinized tissue apical to recession defect [8]. In 2005 zucchelli et al. [9] conducted a long term case series (5 years) for treatment of Multiple Adjacent Recession Defects (MRTD) using envelope type coronally advanced flap and the 5 year follow up showed, increased in keratinized tissue and 85% of treated recessions defects showed complete root coverage. Chambrone et al. [10] conducted a systematic review of periodontal plastic surgery in the treatment of multiple recession type defects and concluded that the mean width of keratinized tissue increased significantly and mean root coverage achieved ranged from 94% - 98% and the need for more randomized controlled trials to identify the indication for each surgical technique. A split mouth study randomized controlled was conducted by pini prato et al. [11] in 2010 to evaluate coronally advanced flap versus connective tissue graft in the treatment of multiple gingival recessions with a 5 year follow up and concluded that 52% sites showed completed root coverage when treated with coronally advanced flap and connective tissue graft in comparison to 35% coverage in coronally advanced flap treated sites. A new minimally invasive treatment of multiple gingival recession defects in maxillary anterior region was achieved by zadeh [12] as a case series by vestibular incision subperiosteal tunnel access technique. The result showed good outcome in esthetic zones.

A randomized controlled trial for treatment of multiple recession defects was conducted by ozcelik et al. [13] using coronally advanced flap combined with orthodontic button application and the result showed 84% complete root coverage and high patient satisfaction with esthetic demands. A novel approach for treatment of multiple recession defects using a Pin Hole Surgical Technique was introduced by John Chao [6]. The Author concluded that 94% mean defect reduction was obtained along with minimum post-operative complications and optimal patient based outcome. The Chao Pinhole Surgical Technique (PST) is a minimally invasive option for treating gum recession. Unlike traditional grafting techniques, PST is incision and suture free. Traditional gum recession treatments involve the use of donor tissue or soft tissue grafts in order to rebuild the gum line. This soft tissue would be sutured in place and would join with existing gum tissue as it healed. During the Chao Pinhole Surgical Technique, a needle is used to make a small hole in the patient’s existing gum tissue. Through this pinhole, special instruments are used to gently loosen the gum tissue. These tools help expand and slide the gum line to cover the exposed root structure. There are no grafts, no sutures, and no incisions needed with the Chao Pinhole Surgical Technique. It simply involves the adjustment of the existing tissue.

Chao conducted different benefits of the Pinhole Surgical Technique

Less discomfort for the patient after treatment.

Faster recovery for the patient than traditional grafting.

No need for scalpels or invasive surgical tools.

No need to take donor tissue from the patient’s palate.

Excellent, natural-looking, long-lasting results.

The Pinhole Surgical Technique, is an incision-free, suturefree procedure for treating gum recession. Through a small pinhole made by a needle and uses specially designed instruments to gently loosen the gum tissue and glide it over the receded part of the tooth. Since there is no incision or suturing, patients can expect minimal post-operative symptoms (pain, swelling and bleeding) [6].

Conclusion

Pinhole surgical technique is one such novel technique which holds promise as a minimally invasive, predictable, efficient, time and cost-effective procedure for recession coverage in Millers Class I and Class II recession defects mainly occur in buccal areas and the main etiologic factors include trauma from tooth brushing, malposition of teeth, ectopic insertion of frenum and muscle attachment. There is a need for more long term research to be carried out to analyze the success of pinhole surgical technique in management of single or multiple recession defects.

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A Summary of Research on Artificial Reefs Monitoring Technology- Juniper Publishers

Abstract

In this paper, the status of oceanic pasture and artificial reefs are introduced. The development of oceanic pasture and artificial reefs monitoring in the word is presented. Development of monitoring technology is divided into three stages according to the history. The first stage is that divers observing the ocean and obtain information about marine organisms. The second stage is using sensor such as sonar, cameras, to observe the ocean and obtain information about marine organisms. The third stage is employing underwater vehicles in the monitoring tasks. Finally we propose that underwater vehicles will be the future trend of artificial reef monitoring based on the advantages of underwater vehicles in monitoring.

Keywords: Oceanic pasture; Artificial reefs; Monitoring; Underwater vehicle; Observing

Status of Artificial Reefs

Currently, Artificial reefs are built around the world: Australia has built Artificial Rocky Reef Fish Communities in Botany Bay, New South Wales [1] French has but artificial reef in golfe juan marine protected area [2]. The United States has built artificial reefs from east and west coasts to the southern gulf of Mexico [3]. Japan's reef construction has a long history over three hundred years. National and local governments invest 60 billion yen per year on the construction [4] China has set up artificial reefs in Zhejiang province, Jiangsu province, Shandong province, Guangdong province, Guangxi province and other places. In order to observe the construction of artificial reefs and growth status of fish in reefs, we commonly use some methods, such as underwater naturalist, underwater photography, sonar, surface buoy and underwater vehicles and so on.

Oceanic Pasture and Artificial Reefs Monitoring at Home and Abroad

Since the complex environment of underwater, it is dangerous for diver to dive in the ocean. Researchers considered a more secure and effective way to monitor ocean. In the second stage, the researchers relied on sensor such as sonar, cameras, to observe the ocean and obtain information about marine organisms. Shyue [4] made research on the distribution of artificial reefs by using multi beam echo sounder. Tian [5] investigated the physical status of artificial reefs by means of a side-scan sonar system. Cuevas [6] introduced the managed Artificial Reefs in Mississippi by using utilizing Side Scan Sonar. Decologne [7] described a technological project to observe submarine that was deployed an autonomous bottom moored acoustic observatory within Bay of Biscay coastal zone. Takahashi [8] made quantitative surveys of fish assemblage at a high-rise artificial fish reef by stationary underwater cameras. Sarria [9] used acoustic communications which placed inside artificial reefs to detect the presence of the species while they are inside. Collins [10] introduced subsea wireless technology which transmitted wirelessly between a seabed sensor and a surface buoy to monitor seabed. Huang [11] calculated indexes of fish resource, through method based on dual-frequency identification sonar. The use of sensor greatly improved the researchers' understanding of artificial reefs and oceanic pastures.

In China related research develops in a slow space. Deng [12] proposed that applying side sweeping to monitor position and height of artificial reefs and Liu [13] calculated the volume of artificial reefs in Dalian by using side sweeping system. Xu [14] proposed that monitoring sea cucumber and sea urchin in the artificial reef of Haizhou Bay by using underwater stereo camera. Liu [15] integrated monitoring and evaluation techniques to apply in the habitat island and artificial reef area. Xing [16] designed a set of video systems for real-time monitoring of oceanic pastures. Shi [17] monitored water quality in the South China Sea by using surface buoy. Wang [18] proposed a complete set of submarine cable online observation system for oceanic pastures which located in weihai xixiakou oceanic pastures to online monitor ecological environment. However, there are no researchers in China designed and manufactured underwater vehicles specifically for artificial reefs monitoring.

Conclusion

In this paper we introduced the development of oceanic pasture and artificial reefs monitoring in detail. This general technology may exist obvious disadvantages in monitoring. We suppose that the underwater vehicles are better tools for monitoring and underwater vehicles will be widely used in future for artificial reefs monitoring.

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Juniper Publishers- Open Access Journal of Environmental Sciences & Natural Resources

Impact of Nitrogen on Phonology and Yield of Rapeseed under Varies Plant Population

Authored by Arshad Ali

Abstract

Competition among plant for same resources such as nutrients, water, light etc ruin its desirable quality when these resources are not in sufficient amount. Fertilizer application and Plant population must be managed properly for satisfactory growth and yield of Rapeseed. A Field Experiment on impact of nitrogen on phonology and yield of rapeseed under varies plant population conducted at Palatoo Research Farm of Agricultural University, Peshawar, Amir Muhammad Khan Campus Mardan in 2015-16. The trail was conducted in Randomized complete block design replicated three times. Three levels of nitrogen i.e. 0, 75, and 125kg ha-1 along with three plant population densities i.e. 10, 15, and 20 plants m-2 were evaluated in the study. Results of the study revealed that seed yield. In short, the plant population of 20 plants m-2 and nitrogen nutrition of 125kg ha-1 were efficiently produced the highest seed yield and.

Keywords: Rapeseed; Nitrogen; Replication; Plant population; Days to maturity; Seed yield

Introduction

Rapeseed (Brassica napusL.) is a member of family cruciferae. It is now the 2nd most important source of vegetable oil in the world and its oil is considered healthy for human nutrition due to moderate content of poly-unsaturated fatty acids and lowest content of saturated fatty acids among vegetable oils [1]. It is source of edible oil production in Pakistan and got attention of people of this country due to its bumper production in the agro-climatic condition of Pakistan. In Pakistan rapeseed was cultivated on 211 thousands hectares with production of 194 thousand tons. Unfortunately the production is decreased 3.2% respectively due to decrease in area [2]. In Pakistan the production of edible oil is not sufficient to fulfill the demand of our country. During the last two and half decades the edible oil demand by Consumer’s has increased from 0.3 million tons to 2.764 tons. Application of nitrogen (N) fertilizer and plant density management regulate rapeseed growth and yield [3]. Rapeseed is very efficient in term of nutrient uptake and require higher amount of nitrogen compare to cereals. Storage of optimum N in stem delayed senescence [4]. Excessive application of fertilizers and leaching of nitrate leads to environmental and economic concern in agriculture, and new target are being set to reduce N supply also improving genetically the NUE of crops including oilseed rape. Suboptimal application of nitrogen suffer protein yield, while oil yield is not favored by excessive nitrogen application [5]. Plant population density influences yield and yield components of rapeseed [6]. Dense plant stand shade the lower leaves in the canopy as effect plant, leading to a reduction in capturing of light [7]. In semiarid conditions, the oilseed crop gave low yield at a plant density of 10-15 plants m-2 [8]. Plant density is one of the most important cultural practices determining seed yield, as well as other important agronomic features of the crop [9,10]. Increments in yield were noted with increase in plant density until other production factors are not limited [11]. Keeping in view the importance of nitrogen and plant population, the trail was conceptualized to optimize production coast of rapeseed with finding of optimum plant stand for higher yield of rapeseed.

Materials and Methods

To study the "impact of nitrogen on phonology and yield of rapeseed under varies plant population" a field research was conducted at Palatoo Research Farm of Agricultural University, Peshawar, Amir Muhammad Khan Campus Mardan during winter 2015-16. Two factors i.e. Nitrogen levels (0, 75 and 125 kg ha-1) and plant densities (10, 15 and 20 plants m-2) were evaluated in the experiment. Seed of rapeseed was sow on a fine and well prepared seed bed with plot size of 2m x1.5 m having 4 rows. A basal dose of P at the rate of 60 kg ha-1 was applied at seedbed preparation. Half dose of Nitrogen (N) was applied at sowing time and remaining half dose was applied at rosette stages. Urea and Single super phosphate was utilized as a source of nitrogen and phosphorus. For maintaining required plant population (10, 15, 20 plants m-2) thinning was done after 20 days of sowing. Irrigation was done according to crop need and weeds were removed manually in each experimental unit.

Data Recording Procedure

Days to emergence were counted from sowing date till 75% plants emerged in a plot. Days to flowering were recorded when 75% plant arise flowers in each plot, days were counted from sowing date till 75% flower formation. Data on pod formation was noted by counting day from sowing up to 80% plant produce pods. Days to physiological maturity were recorded when 75% pods change color and days were counted from sowing to that date. Two central rows were harvested threshed and seeds collected were weighed using electrical balance and converted to kg ha-1.

Statistical Analysis

The data recorded was analysed statistically using analysis of variance techniques (ANOVA) suitable for randomized complete block design. Treatments means were separated using LSD value at 0.05% probability level for significant F test.

Results

The phonological study (days to emergence, flowering, pod formation and maturity) of rapeseed is greatly concerned with maintaining of optimum plant stand and management of nitrogen. Data in Table 1 regarding days to emergence revealed non-significant impact of nitrogen and plant population. Nitrogen and plant population significantly alter flowering duration of rapeseed (Table 1), the interaction between density and nitrogen was found significant (Figure 1). Flowering duration extend with increment in N application. Maximum days to flowering (113.33 d) were taken by the plots fertilized with 125kg N ha-1. Plant population densities had adverse effect on flowering formation. Plant density of 10 plants m-2 took more days to flowers formation. Nitrogen nutrition and various plant population densities significantly affect days to pod formation (Table 1). Days to pod formation increases with increments in N application, while the plant population of 10 plants m-2 take more days to produce pods. Similarly days to physiological maturity was altered by nitrogen levels and plant population densities (Table 1).

Means of the same category followed by different latters are significantly different from each other using LSD test at 0.05 level of probability.

NS = Non-significant, * = Significant at 0.05 level of probability.

The combine effect of nitrogen and plant population density was found non-significant for maturity. The duration of crop extend as the N application is increases. Maximum days to maturity (164.56 d) were noticed in the plots applied with 125 kg N ha-1, while minimum days to maturity (156.33 d) were observed with 0 kg N ha-1. Among different population density, plant population of 20 plants m-2 took more days to maturity. It was observed that decreasing space between plants with increasing nitrogen levels delayed days to physiological maturity (Figure 2). Seed yield of rapeseed significantly affected by plant population and nitrogen levels, whereas the interaction between density and nitrogen was also found significant (Table 2). Plant density of 20 plants m-2 produces the highest seed yield as compared to other plant population. Increments in nitrogen fertilization boast the seed yield of rapeseed. Nitrogen nutrition at the rate of 125kg ha-1 gives higher seed yield (1567.67kg ha-1) than 0 kg N ha-1(1079.67kg ha-1). It was noticed that diminishing space between plants with increasing nitrogen levels enhanced seed yield (Figure 3).

ns = Non-significant, * = Significant at 0.05 level of probability.

Discussion

Days to emergence is not significantly altered by nitrogen and plant population. The non-significant impact of nitrogen and plant population on days to emergence could be explained by the fact that during germination the young seedling utilizes its own endospermic food and not capable of nutrient uptake. Flowers formation is significantly affected by nitrogen and plant population. Delayed flowers appearance was observed with higher nitrogen. It may be due to the fact that higher nitrogen rate enhance the vegetative growth of the plants due to non-stop cell division which ultimately delay the reproductive cycle [12]. Reported that crop phonology of Brassica napus was not affected by N application, but slightly delayed with increasing nitrogen however. With wider plant spacing delayed reproductive growth was noticed in the study. Similarly, reported that increasing the spacing between rows considerably delayed the flowering of rapeseed [13]. The pod formation period is significantly increased with nitrogen nutrition and with 10 plants m-2. This may be due to the sequence of growth stages followed by the crop, as flowering formation is delayed which ultimately extend pod formation period in same pattern. Plant population and nitrogen significantly affect crop duration. Higher nitrogen nutrition delayed crop maturity. Possible reason might be with increasing nitrogen increased the vegetative growth of the plants therefore it delayed the maturity of the crop. Similar results were previously reported by observed that increasing nitrogen nutrition delayed the maturity of the crop [12]. Similarly dense population of 20 plants m-2 takes more days to reach maturity. Likewise increasing seeding rate decreased maturity of canola was earlier reported by [14]. Seed yield of rapeseed enhanced with nitrogen nutrition of 125 kg ha-1 with using of dense plant population. The possible reason might be due to increase in nitrogen and population density pod m-2 increase which cause boast in seed yield. The higher seed yield may be due to efficient utilization of nitrogen by higher plant densities. Present result supports the previous study of [15]. Similarly, indicating that increasing nitrogen and plant population had a positive impact on seed yield [16,17].

Conclusion

From the results discussed it is concluded that nitrogen at 125kg ha-1 with a plant population of 20 plants m-2 perform well in the existing climatic condition and recommended for farmer cultivation in the agro-ecological condition of Mardan.

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Synthesis and Characterization of Low-Cost Activated Carbons for Pollutants Removal from Automotive Emissions-JuniperPublishers

Journal of Chemistry-JuniperPublishers

                                       Abstract

Air purification is one of the most widely known environmental applications of activated carbons. In order to guarantee the successful removal of contaminants and pollutants on activated carbons, the development of new adsorbents has been increasing in the last few years. This paper presents a systematic study for cleaning vehicles emissions of CO, SO2, NO2 and H2S using the process of physical adsorption on novel adsorbents obtained from tropical biomasses. Use of this simple method is a valuable alternative to meet emission standards in Developing Countries. It is known that the agricultural wastes studied here are a feasible alternative for granular activated carbons preparation for pollutants removal during engines operation, approaching its efficiency to the commercial Catalytic Converters.

Keywords: Combustion gases purification; Activated carbons; Adsorption and adsorbents; Pollutants removal

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Introduction

“Quality Can Be Planned.”-Joseph Juran

Wastes cannot be introduced to the environment in unlimited amounts, especially in case of air pollutants. Different measures have been taken to limit the pollution emission. These are e.g.: the elimination of technological processes generating a lot of waste, introduction of new technologies which minimize the contamination, etc. If it is not possible to reduce the emissions, the waste gas must be purified [1,2]. Nowadays, the economic conditions of Developing Countries don’t allow that all individuals own a new automotive. It is necessary to develop alternatives to reduce the negative environmental impact associated with obsolete engines operation. The best way to address it is by reducing certain exhaust gas components during fuel combustion. The answer therefore is to look at vehicles as an integral whole to identify which solutions would be more feasible. Taking this holistic approach to vehicle improvement as a basis, three main exhaust emission control strategies can be defined:

a.Reduction of fuel consumption;

b.Exhaust gas treatment, and

c.Performance monitoring.

From these three alternatives the second one is currently the more effective for air quality improvement. The main gas treatment currently used is the Catalytic Converter, typically comprises of an expensive porous ceramic substrate with large surface area [3]. Unfortunately some users in Central America and the Caribbe an Countries tend to remove the Catalytic Converter from the vehicles to get better power loads. Over the last decade, the study of combustion gas treatment has been focused on more sophisticated Catalytic Converters. Consequently, the study of other alternatives for exhaust gas purification is important. There are a few methods to purify harmful combustion gases such as physical adsorption [4,5], chemical absorption [6], catalytic methods, etc. [7,8]. It is necessary then to select a suitable method to purify harmful gas for Developing Countries. Besides, standards for vehicles become more mandatory day to day. The more feasible alternative would be the development of customized activated carbons filters for the betterment of the environment. This can be accomplished by reducing the emissions that contribute to smog and acid rains [9].

Activated carbons can be obtained from different precursors, with benefits to the environment [10-12]. Due to its chemical composition, forest biomasses are valuable sources in the synthesis of adsorbents materials. Several examples of activated carbons preparation can be found in the open literature [13,14]. They have been used among others in the purification of pollutants gases such as carbon dioxide, sulfur dioxide, hydrogen sulfide, nitrogen oxides and mercury [15-17]. Taking into account this background, the main objectives of this work can be summarized as follows:

I.Study the feasibility of some agricultural wastes as raw material for activated carbons production with high specific surface area, high mechanical resistance and wide availability.

II. Definition of the best experimental conditions for “chemical activation” with H3PO4 such as “physical activation” with steam water for each precursor.

III. Study of the elimination of pollutant gases in vehicles engines with filters of the adsorbents produced.

IV. Proposal of a methodology for filters evaluation in the removal of undesirable pollutants (CO, SO2, NO2 and H2S) during engines operation.

The practical aspects addressed in this research cover the broad spectrum of applications for air cleaning. Better engines performances can be obtained with an adsorption technique of activated carbons, through an extremely economic method.

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Materials and Methods

The raw materials selected for the study are presented in Tables 1-3.

Preparation of Activated Carbons

The starting raw materials were cut up in small pieces and next subjected to pyrolysis. This process was carried out in a tubular reactor in nitrogen atmosphere. The samples were heated (10 °C/ min) from room temperature to the final pyrolysis temperature of 500. In the final pyrolysis temperature, samples were kept for 60 minutes and then cooled down. The solid products of pyrolysis were next subjected to physical activation [18]. In the case of chemical activation the raw materials were the original precursors which were overnight impregnated into H3PO4 and later submitted to pyrolysis into a stainless steel reactor of 30cm of length per 3cm of diameter. Once the reactor reached the desired temperature the samples were kept at the final temperature according to the experimental conditions of the specific experiment. The activated products then cooled down and washed with enough water till get a neutral pH. Finally, the products were dry at 120°C and then stored [19]. Two different processes were used for the synthesis of the activated carbons from the chars previously obtained by pyrolysis. [18,19]. The experimental conditions used were:

Factorial experimental designs 32 were executed to evaluate the simultaneous influence of the activation conditions on the final product features [14,20]. Following the details for both synthesis processes:

Key properties of the activated carbons prepared were analyzed:

a. Raw material availability;

b. High specific surface area;

c. High mechanical resistance;

d. High adsorption speed.

Characterization of the Raw Materials and Synthetized Activated Carbons

Elemental Analysis: The amount of elements (carbon, hydrogen, nitrogen and oxygen) in the raw materials was determined by an Elemental Analyzer by flash combustion. The samples were firstly dried in an oven at 110°C before the measurement was carried out. The materials was burned at a temperature of 1000°C in flowing oxygen for C, H and N analysis in the analyzer. The CO2, H2O and NOx combustion gases were passed through a reduction tube with helium as the carrier gas to convert the NOx nitrogen oxides into N2 and bind the free oxygen. The CO2 and H2O were measured by selective IR detector. After corresponding absorption of these gases, the content of the remaining nitrogen was determined by thermal conductivity detection. The oxygen was calculated by the difference of carbon, hydrogen and nitrogen.

Apparent Density Measurement: Apparent Density is a measure of the mass per unit volume of a material. It is also called Bulk Density and provides a measure of the “fluffiness” of a material in its natural form. In this work the Standard ASTM D1895 was used. According to this standard the materials are poured into a cylinder of known volume (e.g. 100 mL pipettes) and later weight. Apparent density was calculated as the mass of material divided by the volume occupied into the cylinder [21].

Specific Surface Area Measurement: In order to examine the structure of the synthetized materials, the measurement of the specific surface area of the activated carbons was carried out by gas adsorption isotherms using a Sorptometer applying BET Model. All samples were degassed at 200°C prior to N2 adsorption measurements. Specific surface area was determined by a multipoint BET method using the adsorption data in the relative pressure range: 0.05-0.3 [22,23].

Mechanical Resistance Measurement: The mechanical resistance of the obtained activated carbons was measured through a simple method. A know mass of the granular material was impacted by six glass balls into a semispherical container of stainless steel. The percentage relation between the fragmented mass retained in a 0.5mm mesh and the initial mass is used to estimate the mechanical resistance [24].

Adsorption Speed Evaluation: The adsorption speed was determined by Arrhenius Equation:

where: dX/dt is speed of the adsorption process studied; α is the residual concentration of the adsorbed; and k'ads the apparent kinetic constant of the adsorption process that can be determined by:

Applying logarithm to (Equation 2) brings the possibility to change an exponential equation into a linear dependence, see below:

Plotting ln k'ads vs 1 the activation energy (EA) and the preexponential factor (k0) of the adsorption process studied can be calculated. In the Results discussion ads k'ads will be refers as K for practical reasons.

Designing Process of Activated Carbon Filters

Figure 1 illustrates the process of activated carbon units customized assembling. These filters are very useful to study pollutant gases elimination in automotive engines with the adsorbents produced [25]. The samples, in the form of granules of 2-5 mm in diameter, were packed into a steel column (length 300 mm and internal diameter 90 mm). The gas was passed through the bed of the adsorbent at 0.50 L/min. The concentration of CO, SO2, NO2 and H2S were monitored using a Gas Chromatograph Equipment with standard TCD detector. The concentrations were calculated by integration of the area above the curves.

Pollutants Monitoring at Laboratory Scale

There is no known method available in the open literature which is capable of simultaneously determining all components of combustion gas evaluated here [2]. So the method developed in this work in an innovative alternative for Developing Countries. In order to determine the suitability of the obtained adsorbents in the elimination of CO, SO2, NO2 and H2S, the pollutants removal rate was determined. Figure 2 show a schematic diagram of the customized laboratory system for pollutants monitoring. The system includes, among others:

a. 6 cylinder automotive engine;

b. Activated carbon filter;

c. Exhaust gas analyzer device (Gas Chromatograph);

d. Computer system for data acquisition and recording, etc.

Gas Monitoring System: A standard gas chromatography was used with the following specs:

i. Detector: TCD;

ii. Carrier: Helium;

iii. Column: Porapak Q and Molecular Sieve 5A;

iv. Oven Temperature: 100°C;

v. Sample volume: 1 ml;

vi. Carrier Flow: 25 ml/min;

vii. Detector Temperature: 120°C

Under the described chromatographic conditions, the four gases could be easily separated and quantified [25,26].

The methodology used starting with the preparation of the calibration gas sample by injecting known volumes of each of the four pure gases (CO, SO2, NO2 and H2S) and balance nitrogen into adequate Gas Sampling Bag through the bag’s rubber septum. One mL of the calibration gas mixture and the combustion gases were analyzed by a GC system. Randomly measures of combustion gases before and after the purification process were made on a similar manner in order to evaluate the removal rate of the undesirable pollutants.

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Results and Discussion

Composition and Physical Properties of Raw Materials and Synthetized Products

Table 4 brings a summary of the chemical composition (elemental analysis), such as some physical properties of the 5 precursors studied. Elemental nitrogen, carbon and hydrogen were determined from the elemental analyzer by flash combustion while oxygen was determined by the difference of these three elements. Table 4 shows that the largest amount of element in the raw materials was carbon (except for Corncob which had smaller amount of carbon than oxygen), followed by oxygen, and the smallest amount was nitrogen. The lower content of carbon for Corncob can be attributed to a higher content of volatiles in the structure, translated into a high porosity for the raw material. All precursors do not contain sulfur in their structure, which is very favorable from the ecological and technological points of view.

dap: apparent density; dr: real density; P: porosity

The activation process increases carbon amount (~20- 40%) after modification. On the contrary, there was a reduction in the oxygen content (~20-30%) after physical and chemical modifications. There will be also a reduction of hydrogen amount. The amount of nitrogen was so small for all materials. In Table 4 it can be observed that the initial porosity of all material, except corncobs, have lower values, below 0.5, it makes these materials adequate for activated carbons preparation through chemical or physical activation. The best products synthetized by each process will be reported in Tables 5 & 6.

S: specific surface; K: apparent kinetic constant of the adsorption process; Y: yield; Mr: mechanical resistance; dap: apparent density.

S: specific surface; K: apparent kinetic constant of the adsorption process; Y: yield; Mr: mechanical resistance; dap: apparent density.

Activated carbon from Central American Mahogany was the more reactive material with a significant porosity development (S = 847 m2/g). Should be noted also that Activated carbon from Mamey Zapote was the best adsorbent (S=940m2/g) and also have the higher mechanical resistance, yield and apparent density, very important for filters durability, but it’s the less available material. Finally it’s necessary to remark that Common Corncobs, a widely available agricultural by-product in Central American Countries, showed the worst results for all adsorbents properties; it can be attributable to the higher porosity of the initial raw material (P = 0.79%).

Table 6 show that activated carbon from Central American Mahogany was again the more reactive material but registered now the higher yield and adsorbent area (S = 832 m2/g). Furthermore, Activated carbon from Mamey Zapote was the second better adsorbent (S = 805 m2/g) and again have the higher mechanical resistance and apparent density. One more time activated carbon from Common Corncobs was the worst adsorbent (S = 470 m2/g). This fact is a consequence of poorly porous structure development during the activation process. Further analysis of the data from Tables 5 & 6 indicates that irrespectively of the variant used, the process of activation leads to further changes in the structure of carbonaceous material. The activated carbons synthetized from different materials studied here differ significantly mainly in the specific surface area development. The adsorbents differ not only in the surface properties but also in their texture and morphology that depend first of all on the variant of activation and the pyrolysis conditions of the initial material. Figure 3 illustrates the differences in specific surface development with both methods and the same materials.

In Figure 3, it can clearly be observed that larger specific surface area developments were achieved with physical activation processes. Those products were the better adsorbents to remove the undesirable pollutants. It also confirms that the factorial experimental designs used are the most suitable to optimize the conditions for activated carbon preparation. Textural parameters significantly affect the adsorption properties of the samples studied. [5] This observation suggests that the functional groups of the surface also have considerable influence on the abilities for combustion gases removal. All adsorbents studied had a rapid decrease in CO, SO2, NO2 and H2S concentration after gases interacted with the corresponding filters. High intensity of these harmful gas reductions at ambient conditions can be the reason for much better adsorption on higher surface area products. This explains the lower efficiency of gases removal by chemically activated carbons. The chemical activation process has the additional disadvantage of the required product washing after preparation which inevitably aggregates additional costs.

Gas Monitoring System

The calibrating gas analysis through the regression equations obtained from triplicate analysis of the gas mixtures at identical concentrations, revealed excellent agreement with the known concentrations. The pollutants monitoring system and analytical method used were effective for the simultaneous analyses of the four toxic combustion gases. Figure 4 shows two examples of 2 chromatograms obtained during the analysis of combustion gases purified with activated carbons from Mamey Zapote physically activated. In this figure it can clearly be observed the significant difference before and after gases interaction with adsorbents that can remove large amount of these undesirable gases with the associated environmental benefits.

The analyses of combustion gases revealed moderate concentrations of H2S and CO but very high concentrations of SO2 and NO2. The most effective adsorbent to remove these gases were the physically activated ones. At the present state of knowledge we can only speculate about the reasons for such poor results obtained from the chemically activated samples. Most probably the reason is the presence of a large number of acidic groups on their surface, in contrast to the physically activated samples, that probably have more basic functional groups present on the surface of the samples. Other chemicals present in the combustion atmospheres did not appear to interfere with the analyses. The chromatographic peaks were well separated and defined and the gases were present in amounts that could be easily determined. An excellent precision with relative standard deviations significantly below 2% were achieved in all gas monitoring analysis. The speed, sensitivity and selectivity of the used method make it suitable for analyzing combustion gas mixture of the four gases studied. Table 7 shows the overall average values of pollutants removal with activated carbons (A.C.) during automotive engines combustion.

In Table 7 it can clearly be observed that SO2 and NO2 amounts monitored are remarkable higher than the average limit values for 24 h. The good news is that the activated carbons studied can efficiently remove about 80% of pollutants in exhaust gases from automotive engines with the added value that the harmful gases concentration goes below the limit values. Figure 5 show the correlation between pollutants removal rate and specific surface area of activated carbons during automotive engines operation. In Figure 5 it can be clearly be observed that higher activated carbons specific surface area translated into higher pollutants removal rates that could be estimated by the equation 4 with a correlation coefficient R2 = 0.995:

A proper choice of the parameters of chemical and physical activation such as temperature, activation time, activates agent, etc., permits getting universal adsorbents showing very good adsorption properties towards such pollutants as SO2, CO, NO2 and H2S, however more studies are needed.

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Conclusion

Agricultural wastes studied here are a feasible alternative for the synthesis of activated carbons for pollutants removal during automotive engines operation. The main features that make these products feasible for the diminishing of automotive engines emission are their high adsorption capacity, approaching its efficiency to the commercial Catalytic Converters such as the cheaper costs and its renewability. Based on these results the granular activated carbons studied, produced in large amounts, are fully exploitable for combustion gases treatment. The complex composition of the flue gas with SO2, CO, NO2 and H2S can be successfully analyzed with good compound separation and repeatability. The method used in this investigation would be also be suitable for combustion toxicology researches and could possibly be easily modified to analyze these gases when they are liberated from biological sources [27].

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Acknowledgement

The author wishes to acknowledge Maria Andrea Camerucci and Ana Lia Cavalieri from Mar del Plata University, Argentina, they provided a crucial help in the experiments of this work.

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Use of Moringa (Moringa Stenopetala) Seed Extract for Removal of Some Anionic Dyes (Direct and Reactive Dyes) in Textile Wastewater - Juniper Publishers

Abstract

With textile wastewater being one of the most sources of pollution containing higher value of colour, BOD, COD and several pollutants, brings serious problem to the ecological environment. This study is particularly focused on evaluating the efficacy of removal of dyes such as reactive dyes, direct dyes, mixture of dye wastes and mixture of industrial and dye wastewaters by the natural absorbent Moringa. The seeds of the Moringa tree contain a coagulant protein that can be used in the treatment of industrial wastewater. The extracts of seeds (coagulant) obtained by two methods viz. simple extraction with distilled water and with saline water have been used for the study. The effect of some operating parameters on coagulation namely pH, coagulant dosage, mixing time, colour removal and turbidity was studied. It was observed that the colour removals in direct red and reactive red dyes are 94.45 and 98.4% respectively with simple extract of seeds done with distilled water and it was in the order of 96.6 and 97.3% respectively with saline extracts of the seed. These values of colour removal of dyes are optimized at 70ml/L of coagulant and pH 10. The mixture of industrial wastewater and dye wastes, colour removal and turbidity removal was 85.8% with simple extract and 53% with saline extract respectively at optimized point. Moringa stenopetala seed has demonstrated to have high removal ability for anionic dyes.

Keywords: Moringa stenopetala; Natural coagulant;Colour removal; Wastewater; Treatment; Coagulation; Flocculation

Introduction

In textile waste water, dyes are considered the most important pollutants. High volume of waste water that is produced by the textile industry causes water pollution. Generally, dyes in waste water from textile and dyestuff industries are difficult to remove. This is because dyes are usually synthetic and have complex aromatic structures which make them more stable consequently they are difficult to biodegrade [1]. The removal of dyes from textile waste water is one of the most environmental challenge [2]. For many years, researchers have been working on ways of removing dyes from wastewater and different procedures have been developed; for example, physical and chemical degradation and adsorption onto materials such activated carbon and, in addition to a large number of other techniques such as Fenton’s oxidation, electrochemical degradation, ozonisation, etc [3]. The most commonly used in the textile industry are chemical methods that use oxidizing agents such as, peroxide of hydrogen, ozone and purification by physicochemical process of coagulation-flocculation, in which chemical compounds are used, the most employed are iron and aluminum salts [4]. This physicochemical process is widely used both in developed countries as in developing, for it easy operation and low cost. Nevertheless, when applied in textile wastewater, it generates large volumes of sewage sludge and the ineffective decoloration of some soluble dyes.

Moringa tree belongs to the family Moringaceae which is shrubs trees cultivated across the whole of the tropical belt including Ethiopia, used for a variety of purposes such as food, medicinal, and others [5]. The seeds of the Moringa tree contain a coagulant protein that can be used in the treatment of industrial wastewater. For drinking water clarification, Moringa seeds is also used as coagulant/flocculent agent due to its high content of a water-soluble cationic protein which able to reduce turbidity. Oil extracted from Moringa can be used for water treatment, for drinking water clarification and it is also used for textile wastewater treatment [6]. The use of natural coagulant is followed in developing countries, as substitution of external chemical coagulants such as aluminum sulfate and ferric chloride. The water -soluble extract of the dry seeds of Moringa is one of the natural coagulants. Moringa is used for water treatment in two different methods, one as a primary source of activated carbon and the second method through seed extraction, and produce a product working as a coagulant/flocculant agent [2].

At low temperature working conditions, the performance of these customary chemical coagulants is dubious and on-going developments have resulted in the introduction of polymerized aluminium coagulants. Owing to its higher superiority and lower consumption of alkalinity, Polyaluminium chloride (PACl) has garnered a growing market. Flocculants such as the more common polyacrylamide, which are organic synthetic polymer available in the market, offers a wider selection of chemical coagulants to cater for the diverse requirements of the individual water treatment plants [7]. Currently, there is an increased interest in the decolorization and decontamination of industrial textile wastewater. Different treatment technologies have been studied in order to solve the problems caused by the toxic substances contained in industrial textile wastewater, such as electrocoagulation, adsorption, photocatalytic process, ozonation, membrane bioreactor and anaerobic/aerobic biological treatment [8]. However, these methods are neither economically nor technologically suitable for large scale use and normally require the combination of two or three methods to achieve an appropriate level of colour removal [9-11]. The various coagulants attempted for colour removal and their efficiencies are listed in Table 1. To overcome the drawbacks of inorganic coagulants and synthetic polymers associated with growing environmental concerns worldwide, there is a need to consider other potential alternatives for textile wastewater treatment in order to minimize environmental damage and improve the wellbeing of human populations. Therefore, researchers have shown considerable interest in the development of natural polymers as coagulants in the recent past [7,12-14].

RTW: Real Textile Wastewater; SS: Suspended Solids; SSP: Surjana Seed Powder; MSP: Maize Seed Powder; TW, Cr: Tannery Waste, Chromium

Plant materials as coagulants offer several advantages over conventional coagulants such as aluminium sulphate as stated below [8].

1. Activity is maintained over a wide range of influent pH values - no pH correction required

2. Natural alkalinity of the raw water is unchanged following coagulation-no addition of alkalinity required

3. Sludge production is greatly reduced and is essentially organic in nature with no aluminium residuals – sludge volumes are reduced by a factor of up to 5.

4. Minimal coagulant dosage requirement

5. Efficiency at low temperature

6. Chemical coagulants are generally more expensive, toxic and with low biodegradability.

Many researchers carried out studies using Moringa seed for water and wastewater treatment on the seed itself and as cake powder. As the studies reported, the Moringa seeds have content of protein (26.50% - 32.00%), fiber (5.80% - 9.29%), ash (5.60% -7.50%), fat (42% - 45%) and moisture contents (8.7% - 9.1%) [15]. It is also stated that Moringa seeds as one of the most effective natural coagulants, applied to transform water constituents into forms that can be separated out physically. Significant quantities of high molecular weight water-soluble proteins present in the seed of Moringa carry a positive charge [16]. When the crushed seeds added to raw water, the protein produces positive charges acting like magnets and attracting the predominantly negatively charged particles (such as clay, silt, bacteria, and other pollutants). Under proper agitation, these bound particulates then grow in size to form the flocs, which may be removed by filtration or left to settle by gravity [17-19]. In developing countries, Moringa seed is considered favorably in terms to reduce the costs of wastewater treatment in comparison with chemical coagulants [17,20-22]. In addition, the sludge produced by Moringa seed as a coagulant is stated to be innocuous and 4-6 times less in volume than the chemical coagulants produced. In the present attempt, a study has been carried out systematically to assess the efficacy of colour removal in textile waste water containing some anionic dyes such as direct and reactive dyes which are most widely used for coloration of cotton, using Moringa stenopetala seed extracts as coagulant.

Materials and Method

Materials

Domestic mill for grinding the seeds in to powder, nylon sieve filter, Whatman filter paper 4, digital pH meter, UV spectrophotometer (Perkin Elmer, Model Lambda 2500), and Moringa seed were used during the experiment work. The HI93703 Turbidity meter was used to measure turbidity as per ISO 7027 Method. The dyes used in this study were Direct Blue/ Red, Reactive Red/ Blue and a mixture of both dyes; such as C.I Direct Red 81 (λmax 497nm), C.I Direct Blue 86 (λmax 346nm), C.I Reactive Blue 19 (λmax 315nm) and C.I Reactive Red 195 (λmax 532 nm)). Finally, all dyes are mixed together simultaneously with themselves (λmax 362nm) and with industrial wastewaters obtained from Bahir Dar Textile Share Company, Bahir Dar, Ethiopia (λmax 333nm). Various chemicals as presented in Table 2 for making the buffer solutions of different pH, Sodium chloride and ethanol for Moringa seed extraction, were used.

Methods

Moringa (M. Stenopetala) seed collection: Moringa seed were collected manually from the dried pods of trees. The seeds were dried in sun light for three dyes. The hull was removed from the seed surface and wings from the kernels after drying. The kernels were ground in to medium fine powder with a domestic mill to achieve solubilization of active ingredients in the seed.

Coagulant extracts: It is possible to use the Moringa seed as coagulant either as seed or extracting the oil from the seed (defatted cake). To enhance effectiveness on wastewater treatment, some studies have recommended the importance of using defatted Moringa seed[23,24]. In line with this recommendation, defatted Moringa seed was prepared for the purpose of this study.Extracts were prepared by two methods such as a simple extract with distilled water and a saline extract with sodium chloride, as explained below.

1. Simple extract

Production process of simple extract coagulant: Grinding of dry seed without shell→Aqueous dissolution of seed in distilled water→stirring→filtration To prepare 1 L of simple extract coagulant in aqueous solution, 50 g of seed powder was dissolved on 1 L of distilled water by mixing vigorously for 45 min in a magnetic stirrer at room temperature. Then, the mixture was filtered twice: once through commercial filter paper on a funnel and once again through a finefiltering system (Whatman filter paper). The result was clear, milklike liquid and was used as coagulant without further purification.

2. Saline extract

Production process of saline extract coagulant: Grinding of dry seed without shell → Dissolution of seed in a solution of 0.5M NaCl → Stirring → Filtration

To prepare 1L of saline extract coagulant in aqueous solution, 50g of powder were dissolved on 1L of NaCl 0.5M solution by mixing vigorously at pH 7 for 45min in a magnetic stirrer at room temperature. Then, the mixture was filtered twice: once through commercial filter paper on a funnel and once again through a finefiltering system (Whatman filter paper). The result was clear, milklike liquid and was used as coagulant without further purification. After, the seeds kernel dried, a domestic mill was used to grind in to fine powder to achieve solubilization of active ingredients in the seed. The extract oil was soaked in 95% ethanol, 100g of the powder in 500ml of ethanol for 45 minutes at room temperature while mixed with the help of stirrer from time to time. To obtain the defatted cake, it was required to filter the solution using filter paper. The remaining solids (pressed cakes) in the filter were then dissolved in water followed by stirring and filtration in the same way. Then the cake was allowed to dry in oven at 40 °C for 24 hours. In that process ethonal got removed from the seed cake powder. Finally, the dried seed powder is stored under room temperature until it is used for coagulation experiments (Figure 1).

a. Yield of Moringa seed in coagulant preparation

The yield is calculated as given in equation (1).

Yield of moringa=

Where Mi is the original weight of Moringa powder; Mf is the Final weight of Moringa powder after filtration (undissolved solids in the solution).

b. Preparation of synthetic Effluent samples

In order to test the coagulants extracts, in a first stage, synthetic samples were prepared in the laboratory using two types of dyes, all are azo type but with a classification of different class; direct and reactive dye; tested individually and as mixtures and with industrial wastewater, and mixture of dyes added to industrial waste water. The dye stock solutions were prepared by dissolving accurately weighed dyes in water to the concentration of 100-300mg/L, then dyeing with 100% cotton fabric sample following the standard procedures was carried out and dye waste water was collected after dyeing. Different concentrations were prepared from collected dye waste (250, 500, 1000ml in beakers) for treating with coagulant.

c. Effluent analysis

Total solids, dissolved solids, suspended solids, BOD5, COD were measured following standard procedure (AMHA, 1995 and Standard methods, 1995) and SPSS Statistical Data Analysis Software was used for analyzing the data.

d. Optimisation of process parameters

In order to optimize the various process parameters, coagulant dosage range of 10-80ml/L, mixing time of 30-45 minutes, pH in the range of 2-12 were used.

e. Colour Measurements

The difference in absorbance before and after treatment measured in UV-VIS spectrophotometer was used for measurements of colour removal. The results are presented as graphs with respect to various process parameters. The percentage removal efficiency of the parameters was calculated using following formula (2).

Colour Removal efficiency=

Where Ao is the Absorbance value before treatment and A is the Absorbance value after treatment.

Go to

Results and Discussion

Yield of extracts

It has been observed that the yield of moringa seed powder dissolved in the solution is 93.4% and 92.5% for simple and saline extracts respectively. There was no significant difference between the extraction methods in terms of yield and it is evident that maximum percentage of Moringa powder is going to be consumed by the water and used wastewater treatment in the studies by dissolution.

Characteristics of Raw Textile Effluent

An initial experiment was carried out to determine the preliminary characteristics of textile effluent for examining the effectiveness of the M. stenopetala as a coagulant. The pH of the effluent was found to be in the range of pH 9 and 11.5 for all dye effluents and industrial wastewater having dyes Table 3. This indicated that the effluent/dye waste from the textile industry has more alkaline in nature. As the measurement shows, the total solids were found at maximum level of 9000ppm and 8000ppm for the two direct dyes Red 81 and Blue 86 respectively. It was decreased to 3575ppm and 3830ppm after treatment with simple extract and 3500 and 2200 in saline extract as shown in Table 4. In addition, the experiment also confirmed that the particles of TDS are higher than TSS in the textile wastewater samples. Furthermore, the experimental result of wastewater sample show COD higher than BOD5 values. This value indicates, textile wastewater contains high amount of non-biodegradables, 3 to 4 times than degradable organic matters. For instance, direct red 81 abd blue 86 have higher value of COD which is decreased to 750 and 190 respectively in simple extract. In the same way, the highest value of BOD5 was found at reactive 195 and blu 19 which was reduced to 180 and 205 respectively in simple extract. The reduction of COD and BOD5 was also observed in saline extract treatment. The treated effluent was characterized and the values of various parameters (pH, Total Dissolved Solids, turbidity, BOD5 and COD) are compared with the raw effluent. The characteristics of raw textile effluent and after treatment with simple and saline extracts are summarized in (Table 3 & 4). One liter of wastewater was used for the treatment from the total solution. All samples have been mixed for 30-45minutes. From the table, it is also clear that less removal of Turbidity, TS, COD and BOD5 for Industrial wastes and the one mixed with dye solutions waste water due to plenty of other components of wastes in industrial wastewater.

The Effects of processing parameters on Coagulation

Effect of coagulant dose

Coagulant dosage is one of the most important parameters that have been considered to determine the optimum condition for the performance of coagulants in coagulation and flocculation. The coagulant dosage indicates the concentration of M. stenopetala seed extract in the water. This difference is important to note since a lot of the seed mass was separated during the filtration step when preparing the extract. Essentially, insufficient dosage or overdosing would result in the poor performance in flocculation. Therefore, it is significant to determine the optimum dosage in order to minimize the dosing cost and sludge formation and also to obtain the optimum performance in treatment. The effect of coagulant doses (10-80ml/L) on the removal of reactive and direct dyes using Moringa stenopetala coagulant and flocculation time is 30-45min is shown in Figure 2-5. This shows that there was continuous removal of these dye colours with increasing coagulant doses up to 70ml/L as shown in (Figure 2 & 3). After 70ml/L, the colour removal decrease that confirms the optimal concentration as 70ml/L. This may be as a result of re-suspension of solids at this concentration. Furthermore, the high concentrations (>25.0mg/L) of the coagulant confer positive charges on the particle surface (a positive zeta potential), thus redispersing the particles.

It is also an assumption that an increase in the coagulant dose may cause a decrease in pH of system. The decrease in pH may be as a result of the hydrolysis of the coagulants. Low pH values of the coagulated system usually may be attributed to the neutralization of the negatively charged surfaces of wastewater colloids, leading to their destabilization by H+ ions. However, acidification of coagulated wastewater may disturb sorption or could increase the solubility of freshly formed sludge. The highest percentage colour removal of these dyes was found to be 98.4, 86, 94.45, 89.3, 90.5 and 85.8% and 97.3, 84.45, 96.6, 84.8, 87.73 and 84.6% for Reactive red 195, Blue19, Direct Red 81, Blue 86, Industrial waste water and the mixture for both simple and saline extract respectively. This confirms that Moringa seeds to have absorbent properties and effective for removal of colour.

Colour removal

After the characterization of the effluent, the prepared coagulants from the Moringa stenopetala seeds were added to the effluent in the form of coagulant to determine the effectiveness of the extract over the textile effluent. The result shows that the extract removes the turbidity and colour from the textile effluent. Removal efficiency of up to 98.4%, for colour, was reached using 70ml Moringa Stenopetala coagulant extract. The use of Moringa seeds has an added advantage over the chemical treatment of water because it is biological and has been reported as edible [25]. All dyes selected for this study, was prepared in the laboratory for treatments and also mixed with industrial wastewater. A dye solution was prepared as required and its pH was measured by using pH meter. One liter of the initial solution was put into two beakers, and various doses of coagulant were added (10, 20, 30, 40, 50, 60, 70 and 80 in ml/L). The beakers were put one by one onto a standard magnetic stirrer. The solution was stirred for 10 min at high speed and then slowly mixed at 60rpm for 30-35min. After that, the solution was left for 45-60min for settling. The supernatant after settling was filtered through a Whatman filter paper (pore size 20–25μm). The reduction in colour concentration was measured at maximum absorbency visible wavelength of each dye solution was measured by using a UV-VIS spectrophotometer. The result of colour removal is shown in (Figure 2 & 3). In industrial wastewater colour removal is less because the effluent contains high content of dyestuff, surfactants and other additives that are generally made up of organic compounds with a complex structure. These wastewaters are collected from different sections having such different compounds and collected in wastewater plant for treatment.

Effect of flocculation time

The time of macrofloc formation (flocculation time) is one of the operating parameters that is given great consideration in any water treatment plant that involves coagulation–flocculation operations (Figure 4 & 5). Presents the effect of flocculation time using different dose of coagulants for removal of reactive dyes, direct dyes and mixture of industrial wastewater and dyes of textile effluents. The consistence increment of removals has been revealed with increasing flocculation time up to 60min, and then it decreased. The optimum flocculation time was found to be 60min. The highest removal of selected dye colours was found to be 98.4, 86, 94.45, 89.3, 90.5 and 81.5% in reactive red, reactive blue, direct red, direct blue mixed dyes and mixture of dyes/and industrial wastewater respectively in simple extract at 60 minute. As well as, for saline extract the maximum colour removal was 97.3, 84.41, 96.6, 84.8 87.73 and 80.3% in reactive red, reactive blue, direct red, direct blue, mixed dyes and mixture of dyes/and industrial wastewater respectively.

Ebeling et al studied that the removal of Turbidity and Soluble Reactive Phosphorus (SRP, orthophosphate) was increasing as settling time increased from 5 to 45min [26]. A series of jar tests were conducted to fetch the effect of settling time (0, 5, 10, 15, 20, 25 and 30min) on the removal efficiencies of BOD5, COD and TSS using low alkalinity wastewater and the results show that the small particles settle out quickly within the first 5min, with little change in the values up to 15min. The differences in BOD5, COD and TSS removal were not significant after 20min of settling as indicated by other studies [27,28]. The result of settling time is given below in (Figure 4 & 5).

Effect of mixing time

Studies were made to find out the effect of variation of the mixing time on the colour removal efficiency. Various mixing time ranging from 15 minutes to 60 minutes were maintained and the results were reported in (Figure 6 & 7). The experimental result shows that there was a continuous removal of colour, turbidity, TDS and TSS while increasing the mixing time from 15 to 45 minutes. Similar results are observed by Patel and Vashi for some other dyes [29]. When the mixing time is short (<45 minutes), the collisions between the coagulants and colloids are not efficient to precipitate suspended solids in wastewater. On the other hand, if mixing time is longer (>45 min) it would lead to an increase in flocs breakage & limit the size of the floc formed. As a result, small size flocs which are not dense to settle down & finally cause the sample to be turbid again. In sum, it is also found that the longer or shorter mixing time would result in the poor performance of the coagulant seed for binding and bridging. There were similar results in experiments made by other researchers [16]. Initially at less contact time (≤15min.) the maximum colour removal efficiency was achieved to only 63.6 and 63.7% for mixed dyes and reactive red respectively in both simple and saline extract methods respectively. But results show that when the reaction time was increased to 45 minutes, the efficiency was nearly 95% and it decreased when time increased was beyond 45minutes [30- 35].

Effect of pH

The aqueous solution of dye wastes was treated by constant concentrations of dose at 70ml/L in dye wastewater of adsorbent for half a day with varying pH (2 -12). The pH was maintained with the help of buffer solutions. (Figure 8 & 9) show the effect of pH of the dye solution on the decolorization percent within the range of (2-12) for both simple and saline extract methods. The results showed that the decolorization reached maximum between pH 8-10 for both the selected dyes waste and industrial wastewater for all dosages. The effect of pH was one of the crucial parameters to determine the optimum leveling order to minimize the dosing cost and to obtain the optimum performance in treatment. pH variation in comparison had a significant effect on the decolorization of reactive dye, direct dye and mixture of them with textile wastewaters by Moringa stenopetala seed extract.

From these figures, we can understand that the highest colour removal was at pH 10 for all dye wastes. As pH increases from 2-10 the colour removal from textile wastewater increases and reach the highest point at pH 10. The removal of dyes are more at higher pH, because the surface of activated coagulants are negatively charged, the decrease in adsorption capacity at the low pH values would be expected as the acidic medium would lead to an increase in hydrogen ion concentration which would then neutralize the negatively charged coagulant surface thereby decreasing the adsorption of the positively charged ions because of reduction in the force of attraction between adsorbate and adsorbent [36-40].

Removal of Turbidity

Varying doses of the seed extract were added to the effluent followed by mixing at optimum time. The mixture was then filtered through filter paper, finally measure the turbidity value by turbidity meter. The result shows that the higher turbidity removal efficiency was obtained in simple extract method in the order of 84.6% and 83.7% in Saline extract methods for reactive blue dye (Reactive Blue 19) with the dosage of 70ml/L as shown in (Figure10 & 11).

Effect of Dye types

Comparative efficiency of colour removal between reactive, direct dyes and mixture of them with industrial wastewater for both the dyes is shown as bar chart in Figure 12. The average colour removal efficiency of reactive dyes is higher than direct dyes because of the fact that the reactive dyes are a colour that can be water dissolved due to the negative charge of the sulphonate group (SO3-) and direct dyes are the molecular structure with planar positive charges that are more than the negative charges and is water soluble. Direct dyes also have sulphonate (SO3-) functionality, but in this case, it is only to improve solubility, as the negative charges on dye and fibre will repel each other. The colour shade affects the efficiency of reactive colour removal natural coagulant. On the other hand, for the mixed dye, the average colour removal is found between reactive and direct dyes, because all reactive and direct dyes are mixed together and to have interchanging of all charges which affect colour removal [41-43].

Conclusion

Seeds of Moringa stenopetala contain materials that are effective as coagulant. Coagulant dose and coagulation pH are important factors influencing the mechanism of coagulation. Depending up on the type of dye, the coagulation process varies. The optimized parameters for the coagulation of textile wastewater using M. stenopetala were pH 10 and dosage of 70ml/L that can result in removal of 98.4 and 94.45% of colour in simple extract and 97.3 and 96.6% in saline extract in both reactive and direct red dyes respectively. Moringa seed can also remove the maximum turbidity of textile dye wastes to the tune of 85%. It could be concluded that natural coagulant aid created a significant impact on the physical treatment of textile wastewater. The colour removal of mixture of industrial wastewater having reactive and direct dye solution was 85.8 and 84.6% in both simple and saline extract respectively because industrial effluent contains high content of dyestuff, surfactants and other additives that are generally made up of organic compounds with a complex structure while it is collected from different sections which use different compounds.

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[grisdidthis]

hood building post #1ost-c0unt

After redoing the whole thing from scratch FOUR WHOLE TIMES because of being unable to settle on a terrain I was happy with, here we see Platform 42 in its final form. The place has come a long way since V1! (And by “come a long way” I mean, of course, “I am a masochist who loves giving herself more work”, but shhhh. It’s okay. It’s alright.)

To recap: P42 started as a hood this tiny, with seven households and no community lots, target population 201 sims. 

Backstory for it is that the world went up in zombies, and the only place where the undead won’t eat your brain is this city in the middle of the sea that’s run by evil scientists. A bunch of other people live there too, but most of them have been exposed to so much fuckery that by now, there’s not a normal person to be found among them. The story I have on standby at @platform42ts2​ (which is undergoing the slowest and most piecemeal of revamps while yours truly learns how to do Photoshop) follows the quest of a bunch of residents, new and old, who are trying to kick out the mad science gang and turn the place into a more wholesome environment. 

The current hood layout and household distribution, depicted below, reflect the aftermath of those shenanigans, because I’m not waiting the five thousand years it will take to finish and publish the whole story in order to start playing this hood properly. (I’ll just wait the two thousand years it will take me to decorate 15 buildings and edit everyone’s relationships and stats. Nyeh.) 

text to go with those numbers (AND to serve as an index for the “look at this decrepit pile of junk I spent a whole week artfully arranging!” posts that will pop up as I work my way through decorating all of these.)

Apartment Lot: Ardeidae Labs - Apt. #1 - Beaker (Circe & Loki) - Apt. #2 - Curious (Lazlo, Vidcund, Pascal, Jenny, Nervous Subject) Apt. #3 - Beaker/Loste (Kristen & Erin), Apt. #4 - McMillan/Muldeck (Dana & David) 

Community Lot: The Open Library - Owned by Caitlin Cartaine.

Community Lot: The Common Market - Owned by Izza Madhavi

Community Lot: Govender Gardens - Owned by Joko Govender

Community Lot: Administration Office

Household: Sala/Starbrook (Rita & Juniper)

Household: Madhavi/Hamadani (Izza, Fatima, Hafeez, Tariq)

Household: Heron/Maple (Philemon, Mace, Beatrice, the Drude)

Household: Grunt/Smith (Tank, Ripp, Buck, Jill)

Household: Von Laar/Madhavi (Lars & Nadeem) 

Household: Govender/Marais (Joko, Maraliese, Imka, Igmar, Alphonse)

Apartment Lot: Community Housing Center - Dumping ground for all those townies who will inevitably start growing on me to the point where I can’t help myself and promote them to playables.

Apartment Lot: The Old Fish Factory - Apt. #1 - Nagarajan (Nasim & Arjun) Apt. #2 - Nagarajan/Kahale (Alona, Sanjay, Kali, Lani) Apt. #3 - Nagarajan/Loner (Devika & Ajay)

Household: Madhavi/Salman (Abdul & Zeena)

Household: Cartaine/Yi (Moina, Caitlin, Eilis, Jiaying, Suyin, Subject 4) 

LOT USED FOR RANDOM STORY SETS 

So the number of households more than doubled, and the number of playable sims went from twenty to close to fifty (if you include the Strange Isle folks, who weren’t named in the list above because they live in a sub-hood.) HEY AT LEAST ALL THE PEOPLE I SET OUT TO MAKE ARE FINISHED AND PACKAGED. (And I’m long overdue on taking and posting some mugshots of the ones I haven’t shared yet BUT OH WELL.) I’ve resigned myself to the fact that this hood will take an ice age to be what I want it to be, so. Fifteen interior decoration hellscapes left to tackle!

(I say, as if making houses I will never be able to afford IRL isn’t the part of this game I live for.)

[juniperpublishersna]

Bioconjugation among Metallopharmaceuticals: A Review-Juniper Publishers

JUNIPER PUBLISHERS-OPEN ACCESS JOURNAL OF DRUG DESIGNING & DEVELOPMENT

Abstract

This review pertains to an effort to notify the importance of metal binding of naturally occurring molecules. Metallopharmaceutical science is a huge discipline of multifarious applications. In due course of design of metallic drugs one has to rely upon biological relevance of the compound. Sometimes the target activity is lost into toxicity. Hence, the association of a biomolecule or modified bio-compound coordinated with metallic system is the essence of bioconjugation and is the need of hour. Bio-conjugated metallic complexes are always praised for better action. Some diseases have been exemplified in this review and a comprehensive way of presentation has been established throughout the text.

Keywords:  Bioconjugation; AD; Diabetes; Cancer; Antioxidant   

Introduction

Bioconjugation is a meticulous chemical strategy to form a stable covalent link between two molecules, at least one of which is a biomolecule. Synthesis of bioconjugates involves a variety of challenges, ranging from the simple and nonspecific use of a fluorescent dye marker to the complex design of antibody drug conjugates. Antibody-drug conjugates such as Brentuximab vedotin and Gemtuzumab ozogamicin are examples of bioconjugation, and are an active area of research in the pharmaceutical industry [1]. A promising strategy to enable the use of metal nuclides in antibody-targeted imaging and therapy is to design molecules that coordinate to the metal ion and preclude its release in-vivo [2].

A necessary prerequisite of any ligand that binds a metal to form a contrast agent is that the resulting contrast agent be stable so as to prevent the loss of the metal and its subsequent accumulation in the body. Other considerations include an ability to reversibly bind water, which in turn increases it contrastability and decreases the dose level required. This ability is clearly important since the interaction between any two nuclear spins through space decreases at a rate equal to the reciprocal of the distance raised to the sixth power [3].

Hence, metals in medicine are used in organic systems for diagnostic and treatment purposes. Inorganic elements are also essential for organic life as cofactors in enzymes called metalloproteins. When metals are scarce or high quantities, equilibrium is set out of balance and must be returned to its natural state via interventional and natural methods. Metals play a vital role in an immense number of extensively differing biological processes. Some of these processes are quite specific in their metal ion requirements, in that only certain metal ions in specified oxidation states can accomplish the necessary catalytic structural requirement (Figure 1) [4].

One of the principal themes of bioinorganic chemistry is the synthesis of metal complexes that have the ability to mimic the functional properties of natural metalloproteins [5,6]. Proteins, some vitamins and enzymes contain metal ions in their structure involving macromolecular ligands. Inorganic and bioinorganic chemistry are the major contributing fields of medical science and human health witnessed by the past half century. Today, metal-containing therapeutics constitutes a multi-billion dollar industry. Recent investigations in bioinorganic chemistry include the use of metal ions as synthetic scaffolds for the preparation of small molecule therapeutics.   

Insulin Mimicry via Metallic Compounds

In a continued interest towards metallopharmaceuticals (Figure 2) Sodium vanadate and derivatives of bismaltolato- oxovanadium (IV) complexes (BMOV) have been reported to lower levels of blood sugar in diabetic patients [7]. In other words it may be said that scientific community is busy with copying a hormone called as insulin to develop an ultimate treatment of diabetes. Recent under trial experiments with Gold and Silver based glucose level stabilizing agents have further unfurled seek for more efficacy [8,9].

Antidiabetic drugs may be either insulin injections which are used in serious cases of diabetes or oral hypoglycemic drugs, and are suitable for most adult patients. Different hypoglycemic drugs are available in market. These drugs may be classified as the following: Sulphonylureas: increase insulin secretion and help to reduce blood glucose levels. But sulphonylurea may cause weight gain, hypoglycemia and allergic reactions. They are contraindicated in case of pregnancy, lactation and diabetes type 1. They act by affecting the pancreatic β-cells stimulates the movement of insulin-containing secretory granules to the cell surface then into circulation. Biguanides (metformin): They prevent production of glucose in the liver, so improve the body's sensitivity to insulin. They may cause temporary nausea and/or diarrhea, loss of appetite and metallic taste. They are contraindicated with kidney or liver diseases and heart problems. Alpha Glucosidase Inhibitor (Acarbose): They may cause diarrhea, gas, constipation, or stomach pain. Hence, the search for more intelligent/efficient antihyperglycemic or antihypoglycemic agents continues. Dissemination of such area of research expects clinically approved use of metal containing compounds for identifying new medicinal agents from throughout the periodic table to be used as antidiabetic and antioxidant tools.   

Biotransformation of Metallic Compounds

Elemental Medicine is nowadays accepted as a rapidly developing field busy with developing novel therapeutic and diagnostic metal complexes. Advances in biotransformation of metal complexes and targeting, with particular reference to platinum anticancer, gold anti-arthritic, and bismuth antiulcer drugs has remained active goal since decades [10,11]. Studies of iron and copper complexes have shown that they can be more active in cell destruction as well as in the inhibition of DNA synthesis, than the uncomplexed organic ligands [12]. Hence, the field of inorganic chemistry in medicine may usefully be divided into two main categories: firstly, ligands as drugs which target metal ions in some form, whether free or protein- bound; and secondly, metal-based drugs and imaging agents where the central metal ion is usually the key feature of the mechanism of action [13,14]. In addition to metal complexes of novel ligands, compounds of metals with already known organic pharmaceuticals like aspirin, paracetamol, metformin, etc. have gained keen interest [15]. It has been seen that their biological relevance increases on complexing with the respective ligands (organic medicinal chelates). Research has shown significant progress in utilization of transition metal complexes as drugs to treat several human diseases like carcinomas, lymphomas, infection control, anti-inflammatory, diabetes, and neurological disorders [16].

Cancer is the second most frequent cause of death in the world. The discovery of antitumor activity of cisplatin began a search for other metal complexes with cytotoxic properties against cancer cells [17]. The instant information regarding anticancer activities of the ten most active metals: arsenic, antimony, bismuth, gold, vanadium, iron, rhodium, titanium, gallium and platinum have been already updated. Despite the efficacy of cancer treatment using cisplatin, the use is still limited due to severe side effects such as neuro-, hepato- and nephro-toxicity and by resistance phenomena [18]. Gold (III)-dithiocarbamato complexes have recently gained increasing attention as potential anticancer agents because of their strong tumor cell growth- inhibitory effects, generally achieved by exploiting non-cisplatin- like mechanisms of action [19].

The potential applications of Mo-based complexes in medicinal chemistry as metallopharmaceuticals in treating diseases such as cancer and tumors [20] indicate the emphasis of significant approach of non-platin anticancer agents. Ruthenium compounds are highly regarded as potential drug candidates. The compounds offer the potential of reduced toxicity and can be tolerated in-vivo. The various oxidation states, different mechanism of action, and the ligand substitution kinetics of ruthenium compounds give them advantages over platinum- based complexes, thereby making them suitable for use in with promising cytotoxic profiles [21]. The role of transition metals as micronutrients as well as co-factors of several metallo- enzymes in living systems further corroborates the rationale behind synthesis and evaluation of novel transition-metal based complexes for their anticancer effects [22]. Future use of substituted organic ligands and their metal complexes would hence bring forth effective anticancer agents and would depend on structural modifications as would afford them better potency against a number of tumors/cancers, together with low toxicity and better solubility.   

Antioxidant Properties of Metal Complexes

An antioxidant is a molecule that inhibits the oxidation of other molecules. Oxidation is a chemical reaction that can produce free radicals, leading to chain reactions that may damage cells. Antioxidants terminate these chain reactions. Transition metal complexes have been shown to possess encouraging antioxidant activities [23]. Co(II), Ni(II), Cu(II) and Mn(II) complexes of 6-bromo-3-(3-(4-chlorophenyl)acryloyl)-2H-chromen-2-one have been recently found to be effective antioxidants [24]. Generally, antioxidant activity of complexes are determined in- vitro by the hydroxyl radical scavenging, DPPH, NO and reducing power methods [25]. The chemical principles of methods based on biological oxidants comprise superoxide radicals scavenging (O2•-); hydroxyl radical scavenging (HO.); hydrogen peroxide scavenging (H2O2); peroxyl radical scavenging (ROO.) and nitric oxide scavenging (NO.) [26]. Among the non-biological testing scavenging of 2, 2-diphenyl-1-picrylhydrazyl radical (DPPH« assay) and scavenging of 2, 2-azinobis-(3-ethylbenzothiazoline- 6-sulphonate) radical cation (ABTS assay) are mostly experimented. Furthermore, thiobarbituric acid reactive substances (TBARS) and protein carbonyl assays have also been the subject of great attention in this context [27,28]. The novel electrochemical approach to antioxidant activity assay based on the reaction with stable radical 2,2'-diphenyl-1-picrylhydrazyl (DPPH) monitored by the rotating disk electrode (RDE) method has been described advantageous in comparison with usual spectrophotometrical assay since it can be applied to colored compounds and in a wide range of concentrations [29].   

Dementia Relevant Metallic Systems

Alzheimer's disease currently affects over 5.4 million Americans with $236 billion spent annually on the direct costs of patient care [30]. Studies on antioxidant drugs would surely open successful doors to treat AD patients. Seeking for potential antioxidants, chemical behavior of Quercetin as antioxidant and metal chelator has become the subject of intense experimental research [31]. Under comparative antioxidant studies of Co(II), Ni(II), Cu(II) and Mn(II) complexes of 6-bromo-3-(3- (4-chlorophenyl)acryloyl)-2H- chromen-2-one Ni(II) complex shows superior antioxidant activity than other complexes [32]. Commonly it is has been observed that metal complexes may serve as better free radical scavengers [33-35] as compared to the respective free ligands. In some cases antioxidant complexes have rendered a well pronounced larvicidal activity [36]. Hence, synthetic chemistry is playing revolutionary role in human beings by synthesizing novel compounds by different techniques [37]. The target of scientific community has been thus to prepare bioactive compounds relevant to anticancer, antioxidant and enzyme inhibition studies at both the in-vitro as well as in-vivo fronts.   

Biomarkers

Biochemical pathways are famously complex and interconnected, so it’s no surprise that depictions of them have to be simplified (Figure 3). Increasingly, molecular and cell biologists have been coming to terms with the fact that it is hard to decide a label for some protein as a green fluorescent protein (GFP) and expect it to carry on as before. Putting a star next to its name on the whiteboard, or renaming it ‘Target-GFP’, doesn't capture what’s really going on. It is very, very hard to observe living systems at the molecular level without perturbing the very things trying to see, but a great deal of effort is now going into trying to minimize these effects [38]. Under the light shed for evaluation of antidiabetic and antioxidant research, besides developing biomarkers treatment strategies have also been the subject of huge interest. The current status of the aimed field in terms of literature survey is discussed below: (Figure 3).   

Diabetes and Bio-Conjugation

With the aim to continue the enthusiastic search of metallopharmaceutical drugs against diabetes [39,40], thiazolidinediones (TZD) have been reported to be effective anti-diabetic agents that improve insulin sensitivity through the activation of the nuclear receptor and adipocyte-specific transcription factor, peroxisome proliferator-activated receptor gamma (PPAR-γ) [41]. Recently it has been found that Selective PPARγ modulators (sPPARγM) retain insulin sensitizing activity but with minimal side effects compared to traditional TZDs agents [42]. A combination of virtual docking, Surface plasmon resonance (SPR)-based binding, luciferase reporter and adipogenesis assays have been suggested to enlighten the interaction mode, affinity and agonistic activity of L312 to PPARγ in-vitro, respectively [43]. The pharmaceutical isoforms having anti-diabetic effect act by improving the biochemical parameters, this effect is probably due to the high content of polyphenolic compounds found in the formulations [44].

In due course of finding a successful antihyperglycemic candidate, metallic compounds like Vanadium complexes have been well demonstrated in streptozotocin-induced (STZ) diabetic rats and was found that that the vanadate and vanadyl forms of vanadium possessed a number of insulin-like effects in various cells [45]. In the current times basic aspect of diabetes including insulin molecular characterization, chemical basis and its secretion, hypoglycemic drugs and their mode of action associated with diabetes are among the main quests being searched [46]. In an approach of comparative antidiabetic studies of isoforms of BMOV having different metallic centres, it has been found that none so far has surpassed bis (maltolato) oxovanadium (IV) (BMOV) for glucose- and lipid-lowering in an orally available formulation [47]. It is hence clear that ligand and metal selection should be meticulously done to formulate efficient antidiabetic compound.

The bioconjugate chemistry of antihyperglycemic metallic complexes have presented worth some results. The conspicuous application of chromium (III)-amino acid complex against nicotinamide-streptozotocin induced diabetic Wistar rats showed that supplementation of Cr(III)-complex in 8 weeks decreased the blood glucose level in range 46.446-79.593% [48]. Similarly, vanadyl (IV) adenine complex has been introduced as a new drug model for the diabetic complications [49]. Therefore it is expected to be worthy if derivatives of biogenic ligands are formed to design a ligand of favourable properties. For instance, zinc metal-organic framework (MOF) synthesized under mild hydrothermal routes using 5-aminotetrazole and methyl-2- amino-4-isonicotinate anionic ligands has been reported to possess a well pronounced in-vivo antidiabetic activity and low in-vitro cell toxicity [50]. With the same effort,

N,N-Dimethylbiguanide hydrochloride complexes of Neodymium introduced as oral glucose-lowering agent to treat non-insulin dependent diabetes mellitus and to act as antioxidant has shown prominent effect of functional group position in the respective ligands [51].

The medical properties of naturally occurring compounds such as chromones, flavonoids and coumarins are expected to enhance when complex with metal ions suggest the importance of bioconjugate chemical drug research. These complexes can be successfully used in the satisfactory treatment of diseases such as diabetes mellitus [52]. In recent years regulation of the enzymatic activity of human aldose reductase (HAR) has been the main focus of investigation, due to its potential therapeutic application in Diabetes mellitus (DM). Docking behaviour of human aldose reductase (HAR) with different ligands namely such as embelin (Figure 4), copper-embelin complex, zinc- embelin complex, vilangin and quercetin evaluated along with their putative binding sites using Discovery Studio Version 3.1 has shown that that vilangin has maximum interaction energy (-48.94kcal/mol) and metformin with the least interaction energy (19.52kcal/mol) as compared to the other investigated ligands [53]. Therefore, it is strongly suggested that such type of study outcomes might provide new insight in understanding these seven ligands, as potential candidates for human aldose reductase (HAR) inhibitory activity & for the prevention of Diabetes mellitus (DM) associate disorders.

Based on combined in-vitro and in-vitro antioxident evaluation of resveratrol (Figure 5) and molecular modeling studies, it has been indicated that ligand-target interactions/ biological activities are largely dependent on enantiomerism of a target compound [54].   

Antioxidant Activity and Bioconjugation

Antioxidant studies are carried out at the cost of various standard methods [55]. Metal dyshomeostasis is known to be linked with numerous diseases such as Alzheimer's and Parkinson’s diseases, cancer, etc. Recent studies have indicated that some of the metallic compounds of certain ligands may be active while some render inactivity when antioxidant activity test was carried out using picryhydrazyl (DPPH) [56]. On one hand polyphenols have been suggested as efficient antioxidant and anti inflammatory candidates [57] and on the other hand their metallic compounds are expected to exhibit enhanced antioxidant activity due to flexible oxidation state of a metallic centre [58]. Nickel complex of the non-steroidal antiinflammatory drug diflunisal (Hdifl) resulted in the additive antioxidant effect of the respective ligand [59].

The antioxidant activity of the ligand, bis(N-(3-methoxy- salicylidene)-4-amino -phenyl)ether (H2L) and its metal complexes Mn(III) and Cu(II) complexes determined by DPPH, superoxide, hydroxyl and ABTS radical scavenging methods in- vitro, suggest that the Cu(II) complex exhibits greater antioxidant activity against DPPH, superoxide, hydroxyl and ABTS radicals than those of the ligand and the Mn(III) complex [60]. The biotin- 8-hydroxyquinoline conjugates and their metal complexes with manganese(II), cobalt(II), nickel(II), copper(II) and zinc(II) have also been well studied for the possible application in oxidative stress [61]. Similar fashion has been observed with the metallic compounds of

p-coumaric acid [62], 2-(3-amino-4, 6-dimethyl-1Hpyrazolo[ 3,4-b]pyridin-1-yl)aceto-hydrazide [63], chromone Schiff base (Figure 6) [64], etc.

Another important aspect of the antioxidant studies is the strength of a complex not to undergo ROS generation to render a mechanistic action without harming a normal mammalian cell e.g., Ag complex of 1, 10-phenanthroline [65] has shown an interesting behaviour in this context.   

Sugar and Urea Derivative Based Complexes

Urea derivatives bonding through the nitrogen, sulfur and oxygen atoms to the central metal ion form an important class of biologically active ligands. They have been receiving considerable attention due to their pharmacological properties, anti tubercular activity, antiviral potentiality, activity against protozoa small pox and certain kinds of tumour [66]. The chelating characters of thiosemicarbazone have been studied very widely with different metal ions, their complexes with transition and non transition elements were reported.

The ability of sugars to sequester metals is of current interest in the possible development of metal chelates for clinical use and as models for biologically important compounds. Amino sugars form Schiff base with salicylaldehyde and other aromatic aldehydes and only few reports of transition metal complexes of these ligands have been found. Metal chelation could be a rational therapeutic approach for interdicting Alzheimer's disease (AD) pathogenesis. Amyloid plaques that are clusters of proteins and metal ions accumulated between neurons (nerve cells) in Alzheimer's patients’ brains. Enhancing the targeting and efficacy of metal-ion chelating agents through sugar appended ligand is a recent strategy in the development of the next generation of metal chelators.   

Conclusion

From the overall survey it has been established that biomolecules impart great effects in metallic systems to develop molecules of interest. Metallopharmaceuticals are engaged in designing heme-oxygenase and nitric oxide synthase models to bring forth highly demanded gasotransmitter efficiency applicable at various bio-essential routes. Under these circumstances scientific community should fabricate bioconjugated systems to form compounds of human beneficial and multi-purposeful.

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