Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241102EnglishN2018January19General SciencesStudy of Biodegradation of Luffa Cylindrica/Poly(lactic) Acid Composites English0105Swarnalata TripathyEnglish Chinmay PradhanEnglish Chhatrapati ParidaEnglishObjective: The main objective of present research is to convert the agricultural waste Luffa cylindrica (LC) fiber into a valuable product. Methodology and Results: By injection molding technique, biodegradable composites were fabricated using 2, 5 and 10 weight percent of natural fiber of LC in biodegradable polymer matrix poly lactic acid (PLA).The patterns of disintegration of the virgin PLA and its composites were studied for a period of 75 days in sand, soil and compost environment and for a period of 30 days in bacterial and fungal conditions. The biodegradation were evaluated by measuring weight loss of the composite samples with time at every interval of 15 days in the above mentioned environment. The probable outcome of the study presents that the PLAand the composites using LC fibers almost degrade completely in bacterial environment after 30 days. However,in fungal environment the degradation was found upto 4 %. Lastly the composite samples were unable to degrade in sand, soil and compost environment. Before reinforcement the LC fibers are treated chemically for enhancing fiber matrix adhesion. The thermal degradation and the structural changes of the chemically treated LC fibers are studied by thermogravimetry analysis and X ray diffractometer. Conclusion: The fiber parts were mostly consumed by microorganisms. High fiber loading provides larger surface area for the microorganisms as a result the degradation rate is found to be high. Thus, the degradation process is accelerated by the microorganisms. EnglishBiodegradation, Biocomposites, Poly(lactic) acid, luffa cylindricaIntroduction Increasing environmental concerns has motivated the researchers worldwide to develop and utilize materials that are compatible with the environment. Natural fibers have gained importance as they are cheaper, environment friendly and more sustainable than synthetic or man-made fibers like glass, kevlar, aramid, nylon, terylene etc. Natural or plant fibers are renewable and when they are burnt they leave very less residue and discharge less carbon dioxide in the environment. Thus composites made from natural fibers are environment viable. In the prevailing research, a common tropical fruit found in Odisha, India. luffa cylindrica (LC) also known as sponge gourd, vegetable sponge, wash sponge, gourd towel, dishcloth gourd, loofah gourd is chosen as reinforcement for fabrication of composite materials.It belongs to the family cucurbitaceae and is abundantly found throughout India as an agricultural waste. The LC fibers are composed of 60% cellulose, 30% hemicellulose and 10% lignin. The cellulose present in plant fiber consists of some identical units named as glucopyranoze unit. Each unit contains three hydroxyl groups, which are the active sites for strong bond formation. When the cellulose percent is high, the number of active sites are high leading to better adhesion between fiber and matrix. Such high percent of cellulose in the LCfiber is the motivating agent of our investigation.PLA is one of the most promising biopolymer which is completely biodegradable. It is obtained from natural feed stocks as well as agricultural wastes. It has a wide range of applications in various fields and possess various advantages such as biocompatibility and better thermal processibility. PLA is hydrophobic and LC fibers are hydrophilic. They are thermodynamically incompatible and it leads to poor fabrication of the composites. As the chemical nature of fiber and matrix are different, various treatments like chemical treatment, electron beam irradiation, gamma irradiation, plasma treatment, ultra violet exposure etc are given to the fiber before reinforcement to improve compatibility between LC fiber and PLA matrix. In this study the LC fibers are exposed to various chemical treatments like alkali treatment, bleaching and acid hydrolysis. Biodegradation is the enzymatic breakdown of living organisms such as bacteria and fungi into their metabolic products. During the process of degradation, PLA gradually degrades to lactic acid which splits into CO2, H2O and biomass.Thermal analysis of composite material is an influential tool for understanding the structural property,heat resistance capacity,mass degradation etc. Thermogravimetric analysis(TGA) is the most important technique to study the thermal behaviour of materials. The information regarding thermal stability of composite materials are indispensible for proper use of these materials. Kumar et alin 2010 investigated biodegradation of flax fiber reinforced PLA composites. They reported that different amphiphilic additives can be added for delaying and The study of biodegradation of natural fiber reinforced polymer composites have been carried out by many researchers. S.N Surip et. al., in 2016 studied the degradation of Poly lactic acid reinforced with kenaf fibers. The study revealed degradation  PLA / kenaf fiber composites increased by 17% compared to degradation of only virgin PLA. A. Araujo et al in 2014 carried out biodegradation assessment of PLA and its nanocomposites in presence of organoclay. They reported that degradation rate of nanocomposites of PLA increased when the composites were exposed to ultra violet radiation. Also they revealed that the rate of degradation was affected by the presence of organoclay in the composites. However, the study of biodegradation of composites involving PLA and LC fibers are scarce in literature survey. The present research deals with extensive study of biodegradation of PLA/LC fiber composites in soil, sand, compost, bacterial and fungal conditions. Research Methodology - Materials- Polylactic acid (PLA) of grade 4042 D (molecular weight MW~6,00,000), was purchasedfrom Nature Works, USA. The LC fiber was collected from the local forest area. All chemicals such as sodium hydroxide (NaOH), sodium hypochlorite (NaClO), sulfuric acid (H2SO4) of AR grade were purchased from E. Merck, India. Chemical treatment of LC fiber- Small pieces of LC fibers were cut of length around 2cm and then were washed thoroughly, in order to remove the impurities such as oil, dust etc. Then these were dried at 700C in vacuum oven for 20 minutes. Chemical treatments like alkaline treatment followed by bleaching and acid hydrolysis were done. The LC fibres were soaked in 5% sodium hydroxide solution at 800C for 1 hour for alkali treatment. The main drawback is hydrophillic nature of the natural fibres which causes poor adhesion between fiber and matrix. Therefore, such composites possess very poor mechanical properties in wet conditions. The LC fibers were treated with sodium hydroxide to improve interfacial bonding and to reduce moisture absorption. The hemicellulose and lignin were treated from the natural fibers during alkaline treatment. The number of hydroxyl groups present in the fiber are gradually reduced in order to increase the hydrophobicity of fibers that helps in strengthening the bond between fiber and matrix. Bleaching of alkali treated LC fibers was done with 2% sodium hypochlorite solution. Continuous stirring of the mixture was done at 800C for 2 hours. Bleaching increases the whiteness of the fibers. The bleached LC fiber/ water suspension was finally prepared andkept on an ice bath. Along with continuous stirring, sulphuric acid was added slowly to the suspension kept in an ice water bath till it reached the final concentration of 60% sulphuric acid. Under continuous stirring the obtained suspension was heated at 250C for 20C. The mixture was centrifuged in an ultracentrifuge for 20 minutes at 300C with 7000 rpm to remove excess acid. Isolation of micro and nano-fibers with a high degree of crystallinity was obtained by acid hydrolysis by removing the amorphous regions of the raw cellulose material. The degree of polymerization (DP) and the molecular weight of the bleached fibers decreases due to acid hydrolysis. Processing and fabrication of composite - Prior to use, the PLA pellets and the LC fibers were dried at 800C for 24 h under vacuum. In a micro-compounding moulding equipment (DSM Micro SSC Compounding System, DSM research, The Netherlands), the polymer and fiber were mixed at 1700C for 10 minutes at 100rpm.  The PLA pellets and LCfibers were mixed In different weight proportion to obtain B0, B1, B2, B3, B4, B5 and B6 samples. The virgin Poly lactic acid is B0. The LC fibers are in the weightratio 2%, 5% and 10% in B1, B2 and B3 samples respectively. In samples B1, B2, B3, the LC fibers are subjected to chemical treatments such as alkali treatment, bleaching and acid hydrolysis. The LC fibers are in the weight ratio 2%, 5% and 10% in B4, B5 and B6 samples respectively. But in B4, B5 and B6 samples, LC fibers were exposed to only alkali treatment and bleaching. Fibers were not subjected to acid hydrolysis. Biodegradation of LC fibers in soil, sand and compost –                                         The degradation of the samples were checked by weight loss over time (at an interval of 15 days) in soil-sand, sand-compost, soil-compost and soil-sand-compost. Samples were put in thermocol cups containing mixture of sand, soil and compost. These cups were kept at room temperature. Water was sprinkled on it every day. After every 15 days, samples were taken out, washed with water and then dried. Then the samples were weighed for evaluation. Microbial degradation of LC fibers - Degradation by bacteria – 8.75 grams of nutrient broth was dissolved in 350 ml of distilled water. It was mixed thoroughly. 7 conical flasks were taken and in each flask, 50ml of nutrient broth was poured. The mouth of the flask was plugged with cotton and covered with brown paper. It was autoclaved for 45 minutes. The conical flasks were kept in laminar air flow under ultra violet light for 20 minutes. Then 20μL of bacterial strain (Bacillus subtilis) was added in each flask using sterilised micropipette and these flasks were kept in an incubator at 30ºC. After 24 hours, the samples of LC fibers were put in the flask. After 15 days the samples were taken out, washed and dried. Then the samples were weighed for evaluation. Degradation by fungus – 350 ml of potato dextrose broth was done by dissolving 7.4 gms of potato dextrose broth powder in 350 ml of distilled water. In 7 conical flasks, potato dextrose broth was poured, each flask containing 50ml of potato dextrose broth. The mouth of the conical flask was plugged with cotton and brown paper was wrapped over it. It was autoclaved for 45 minutes. The conical flasks were then placed in laminar air flow under ultra violet light for 20 minutes. Then 20μL of fungal strain (Fusariumoxysporum) was added in each conical flask and these flasks were kept in an incubator at 37ºC. After 72 hours, the samples were put in the flasks. Result – Biodegradation of composites Biodegradation studies of neat pure PLA and PLA/ LC fiber composites were carried out in soil/sand, sand/compost, soil/compost, soil/sand/compost, and finally in bacterial and fungal environments. Ultimately, biodegradation depends on the nature of the materials. PLA biocomposites will expand further if the materials degrade faster after their end-uses. Biodegradation of composites in soil/sand  The PLA and its composites are not affected by soil/sand composition which may be attributed to chemical nature of PLA.The degradation nature is closely related to the chemical component and bonding characteristics of the plastic.   Discussion The figure 1 shows intense peaks at 15.87o and 22.86o. The peak at 15.87o corresponds to amorphous cellulose or cellulose II of [101] crystallographic plane. The peak at 22.86o correspond to crystalline cellulose [cellulose I] of[002] crystallographic plane. Natural fibers are composed of cellulose, hemicelluloses and lignin.The presence of peaks at 15.87o and 22.86o confirms that when the LC fibers are treated chemically, hemicelluloses and lignin are removed and only cellulose is present after chemical treatment. Two different crystalline allomorphs have been identified by the characteristic X-ray diffraction (XRD) patterns of treated fiber viz. cellulose I and cellulose II. Cellulose I is the most abundant form found in nature. Cellulose II can be prepared by mercerization or alkali treatment.In other words, the chemical modification of LC fiber exposed large numbers of cellulosic hydroxyl groups which can increase the adhesion between the fiber and matrix. Removal of hemicelluloses and lignin decreases the hydrophilic nature of the fiber which attributes for increase of bio compatibility between fiber and matrix. Natural fibers are hydrophilic in nature and absorb a lot of moisture. Thus when the fibers are heated there is loss of mass due to the moisture in the temperature range of 150oC. But when the LC fibers are treated with alkali, the hydrophilic nature is decreased due to removal of hemicellulose and lignin. This is evident from figure 2. When the chemically treated LC fiber is heated up to 200oC, there is only a marginal loss of mass of 2.37%. This is shown as stage I in the figure 2. In the second stage (II) in the temperature range 250oC to 350oC a rapid loss of mass of around 55% is observed. This loss of mass is due to degradation of amorphous cellulose present in the sample. In the third stage (III) beyond 350oC the degradation is due to decomposition of crystalline cellulose. There is a mass loss of 76% observed for treated LC fibers at 600oC compared to loss of only 61% in the composites with untreated LC fibers. The higher loss of mass of the cellulose fibers at high temperatures beyond 350oC may be attributed to removal of lignin and hemicellulose during the chemical treatment. Figure 3 shows composite degradation in soil and compost. Here the sand was replaced by compost which initiated the bio degradation. Composites B1 (2%fiber) and B2 (5%fiber) samples degraded slightly. No biodegradation was seen in other composite samples. The moisture and heat present in the compost attacks the PLA chains and splits them apart leading to smaller polymer fragments, and finally, lactic acid. It can be suggested that microorganisms found in active compost consume the smaller polymer fragments and lactic acid as energy source. PLA degradability is driven by the hydrolysis and cleavage of the ester linkages in the polymer backbone. But when the fiber weight percentage increases as in composite sample B3 (10% wt reinforcement) biodegradation didn’t occur. It may be due to non-breakage of polymer chain by presence of excess fiber. Also no biodegradation was observed in composite sample B4, B5 and B6 where the fiber were exposed to only alkali treatment and bleaching. So it can be concluded that the fiber treated with alkali followed by bleaching and acid hydrolysis (sample B1, B2)are actively biodegraded. Figure 4 shows degradation in sand-soil-compost medium. Composite sample B0 (virgin PLA) does not degrade at all. However when the virgin PLA is reinforced by LC fibers the composite samples start degrading.Thus soil /compost/sand environment is further a better active medium for biodegradation as compared to sand/compost and soil/compost environment. Figure 5 indicates degradation of composites in bacterial condition. Allthe composite samples degraded in bacterial environment unlike soil/ sand/compost environment where only composite samples B1 and B2 degraded. Thus the weight of LC fiber in composites play an important role in bio degradation in bacterial environment. However the rate of degradation in sample B4, B5 and B6 are observed to be more compared to sample B1, sample B2 and sample B3.Thus when the fibers are subjected to acid hydrolysis as in  sample B1, sample B2 and sample B3  the degradation is low. This may be due to strong bonding between acid hydrolyzed fiber and PLA matrix. Thus degradation in bacterial environment is more prominent compared to soil/ compost/sand environment. Figure 6 shows degradation of composites in fungal condition.  Again the rate of degradation in sample B4, B5 and B6 are observed to be more compared to sample B1, sample B2 and sample B3 which shows better chemical bonding between acid hydrolyzed fiber and PLA matrix. However degradation in bacterial environment is more prominent compared to degradation in fungal environment. Conclusion The biodegradation of PLA/ LC fibers were carried out in sand, soil, compost and microbial (bacterial and fungi) conditions at room temperature. The composite did not degrade in sand, soil and compost. But the PLA composites degraded under bacterial and fungal conditions. This work effectively demonstrates the conversion of a waste product like LC fiber into a highly valuable product and its disposal under various environments. Acknowledgement Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Major research project funded by DST, Government of Odisha, 1184/ST-(BIO)-02/2017 Dt: 1/3/17 Englishhttp://ijcrr.com/abstract.php?article_id=2420http://ijcrr.com/article_html.php?did=2420 AraújoA., OliveiraM., Oliveira R., Botelho G., Machado A. V. ; `Biodegradation assessment of PLA and its nanocomposites’; Environ SciPollut Res (2014) 21:9477–9486 Bhatia, A, Gupta, R, Bhattacharya, S, Choi, H. International Polymer Processing, vol.25 (1), 5-14(2010). Kumar R., Yakubu M.K., Anandjiwala R.D.; `Biodegradation of Flax Fiber Reinforced Poly lactic acid’; Express Polymer Letters Vol.4, No.7 (2010) 423-430 S.N. Surip, W.N R Wan Jaafar, N.N. Azmi, N.A. Hassan.; `Biodegradation properties of polylactic acid reinforced kenaffibers’; Acta Physica Polonica A Vol.129, No.3 (2016) 835-837

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241102EnglishN2018January19General SciencesFormulation of Cost Effective Alternative Bacterial Culture Media Using Fruit and Vegetables Waste English0615Pratibha JadhavEnglish Mrunalini SonneEnglish Arati KadamEnglish Suraj PatilEnglish Kirti DahigaonkarEnglish Jaspal Kaur OberoiEnglishVegetable stalks and fruit peels are generally used for composting or merely disposed off as waste. Considering their nutritional values, these kitchen wastes can be utilized for production of alternative cultivation media. Cost of conventional culture media is very high. Use of these alternatives would reduce the cost drastically. The present study is aimed at replacing the nutrient source by various locally available cheap materials, such as vegetables and fruits waste, that contains considerable amount of protein and starch. These raw materials are cheap and easily available in local shops and vegetable markets and also as kitchen wastes. They were selected as a natural nutrient source to prepare the alternative culture media. Growth of bacteria and pigment production efficiency was analyzed using nine formulations. The results showed that drumstick  formulations having seed and peel extract, formulations B and D supported growth of E.coli, Serratia sp., Pseudomonas sp. EnglishAlternative media, Cost effective media, EDS analysisINTRODUCTION Microbiological studies depend on the ability to cultivate and maintain microorganisms under laboratory conditions by providing suitable culture media that offers favorable conditions. A nutrient media prepared for the growth of microorganisms in a laboratory is called culture media. Microorganisms can obtain energy directly from sunlight while carbon can be made available in organic forms such as carbohydrates or inorganic forms such as carbon dioxide and water. Nutrient agar medium is commonly used as general purpose medium for the cultivation of broad range of bacteria. It is a basic medium composed of peptic digest of animal tissue, beef extract and yeast extract, sodium chloride and agar. Commercially available media such Nutrient Agar, Cetrimide Agar, MacConkey Agar are used for the growth of microorganisms but these are very expensive. In today’s world waste disposal is also a major problem. So lot many researches are carried so as to use domestic waste for production of cheap media. Higher cost of cultivation media is a matter of concern[2]. Therefore, various alternative media are formulated and alternatives for agar are tested, so as to reduce the cost involved. Agar is a solidifying agent and very few studies have concentrated on replacing agar for solidification. By comparing other studies carried out in this area by a number of researchers, now it would be possible to use a number of sources as alternative culture media[1]. Even then, increasing cost of culture media has necessitated continuous search for more readily available culture media at affordable price. Different media for the growth and isolation of organisms have been reported from different substrates[6]. Some vegetables and fruits have been used to cultivate both fungi and bacteria, such as Gooseberry[36],Carrot, Tomato, Cabbage, Pumpkin etc [8]. with easily available low cost material as substitutes for Nutrient Agar. Some others have used cow pea, green gram and black gram as starch and protein substitutes to reduce the cost of microbial media[40].               Microorganisms are omnipresent and very diverse. Preparation of suitable culture media is one of the necessary to study them. Different microorganisms grow in different environments and have variety of growth requirements; like nutrients, pH, osmotic conditions and temperature[7]. The current limitations of cultivation of microbes in lab need to be addressed by formulation of newer media. Microbial culture media can be of different types, depending on the nutritional growth requirements of the microorganisms. Microorganisms require various macro elements namely C, H, O, N, S, P, K, Ca, Mg and Fe. For Carbohydrates, Lipids, Proteins and Nucleic acids synthesis C, H, O,N S, P are used, while K, Ca, Mg, Fe exist in the cell as cations, playing a variety of roles. In addition to macro elements, all microorganisms require several microelements like Mn, Zn, Co, Mo, Ni and Cu. These are generally part of enzymes and cofactors. Microorganisms also require organic compounds as growth factors. MATERIAL AND METHODS Collection of samples              Vegetables and fruits like Drum stick (seeds and peels), Orange peel, Potato peel, Cauliflower stalk and Fenugreek stem were collected from local vegetable market. The collected samples were transported to the laboratory and processed immediately. Treatment of samples           Peels, stalks and seeds were sun dried for 2-3 days. Dried material was grinded to powder using electronic blender. The powdered samples were kept in air tight containers until its use. Test organisms used E.coli,  Serratia sp., Pseudomonas sp. Formulation of Media            The dry powder was kept in warm water for 2-3 hours. Then filtered with the help of filter paper and filtrate was used to nine different solid formulated media. Then agar, which is solidifying agent was added in 100 ml distilled water[9]. In all experiments pH of the media was adjusted to 6.5 - 7.0. The dissolved media was sterilized in autoclave (Dixons, ST19T) at 121°C for 20 minutes under 15 psi pressure and were poured into sterile Petri dishes separately. Preparation of fresh culture Bacterial cultures used for analysis were E.coli, Pseudomonas sp., Serratia sp.. Then these bacteria were streaked on the freshly prepared NA medium from the original stock culture. The cultures were allowed to incubate at 37°C for 24 hours. Microbial inoculation into alternative media The young cultures of test bacteria such as E.coli, Pseudomonas sp., Serratia sp.. were inoculated in triplicates in each alternative culture medium. Then all the plates were incubated at 37°C for 48 hours. After the incubation all the plates were observed for bacterial growth and pigmentation. Analysis of bacterial growth in formulated media Bacterial growth was checked at 37°C for total duration of 24 hrs. Time interval selected for growth curve was one hour. Fresh culture of test organisms was inoculated in Nutrient broth and formulated medium. Growth was measured at 600nm. Estimation of proteins and carbohydrates Protein was estimated by Folin Lowry’s method while carbohydrates content was analyzed by DNSA method. Chemical analysis of dehydrated powder The chemical composition, macronutrient content of the tested samples was determined by EDS (Energy Dispersive Spectroscopy) analysis. RESULTS Alternative media supported the growth of test organisms like E.coli, Pseudomonas sp., Serratia sp. There was no significant variation in the colony morphology. Pigmentation was slightly affected in some cases. Growth of Serratia sp. on different media Serratia sp. grown on control (NA) showed pigmentation. Serratia sp. grown on formulated media showed no significant variation in colony morphology. Serratia sp. showed maximum growth and pigmentation on DP, DS, B, D formulations. Pigmentation was slightly affected in A, E, F formulations. Growth of Pseudomonas  sp. on different media Pseudomonas sp.  grown on control (NA) showed pigmentation. When Pseudomonas sp. was grown on formulated media there was no significant variation in colony morphology but showed no pigmentation on DP, DS, A, B, C, D, E, F formulations. Growth of E.coli sp. on different media E.coli grown on control (NA) showed growth. When E.coli was grown on all formulated media there was no significant variation in colony morphology. Fig  3 Growth of E.coli on different media:   Microbial growth curve:                                       The microbial growth was tested and measured at different incubation intervals. Growth was measured at 600 nm on spectrophotometer. When growth of Serratia sp. in NA was compared with DP,DS,A,B,C,D,E,F, it was found that maximum growth was found in DP,DS,B and D and less growth was found in A,C,E,F,DS. Percent increase in growth in comparison to NA was DP-25%, B-42%, D-33%. When growth of E.coli sp.in NA was compared with DP,DS,A,B,C,D,E,F, it was found That max growth was found in DP, DS, B and D and less growth was found in A,C,E,F,DS. Percent increase in growth in comparison to NA was DP-16%, B- 20.75%, D-14%. When growth of Pseudomonas sp. in NA was compared with DP,DS,A,B,C,D,E,F, it was found that maximum growth was found in DP, DS, B, and D and less growth was found in A,C,E,F,DS. Percent increase in growth in comparison to NA was DP-8 %, B-47.61%, D-33% Protein and Sugar concentration was estimated by standard method. B media contains 10 mg/100 ml of Sugar and 8.7 mg/100 ml of protein. D media contains was 9.9 mg/100ml of Sugar and 8.9 mg/100 ml of protein. DP media it was 0.7 mg/100ml of Sugar and 6.3 mg/100 ml of protein. Thus, being rich in sugar and proteins, the media is able to support growth of microorganisms. Chemical analysis of the dehydrated powder: To determine chemical composition and nutritional content of formulated media EDS (Electronic Dispersive Spectroscopy) analysis was done. In B formulation Magnesium (Mg), Molybdenun(Mo), Nikel(Ni) was found which was absent in control NB. But Cu (copper), Chloride(Cl), Sulfur(S) are present in control NB and absent in B formulation media. In D formulation Magnesium (Mg), Calcium (Ca), N (Nitrogen), was found which was absent in control NB. Cu (copper), Sulfur (S) was present in control NB but absent in D formulation. In DP formulation Magnesium (Mg), Phosphorus (Ca), N (Nitrogen), was found which was absent in control NB. Cu (copper), Aluminium (Al) was present in control NB but absent in formulated DP media. Comparative cost study of media: The cost of Alternative media is drastically less than the commercial liquid media. The commercial liquid media containing agar for solidification is also very cost effective as compared to commercial solid media               The cost of alternative media is drastically less than commercial liquid media. The B, D, and DP media containing agar for solidification is also very cost effective as compared to commercial solid media. The rise in cost of B, D and DP agar media is due to incorporation of agar which adds on to the cost Alternative cheap source of solidifying agent needs to be found so that this cost be brought down. DISCUSSION                Cultivation of microorganisms in laboratory requires nutrient environment which serves as source of nutrients for growth. Our investigation was aimed at replacing nutrient media by alternatives such as vegetable and fruit waste. In reported studies; carrot, tomato, cabbage, pumpkin waste was used in formulation of media[8] but in our research we introduced cauliflower, drum stick, orange peel, potato peel and fenugreek stem for formulation of media. CONCLUSION The formulated medias A to F supported growth of bacteria such as E.coli, Pseudomonas sp., Serratia sp. In preliminary study it was found that E. coli grew well in B formulation in 24 hours.  Pseudomonas sp. grew best in B formulation in 24 hours while pigment production was observed after 48 hours. Serratia grew best in all formulations and pigment production was observed in 24 hours at room temperature. EDS analysis was done to find the elements present in NA, DP, B and D formulation that supported growth of  E.coli, Serratia sp. and Pseudomonas sp. After comparing the growth of organisms in commercial and alternative media, the growth of Serratia sp on alternative media shows 25%, 42%, 33% rise in DP, B and D media respectively. E.coli shows 16%, 21%, 14% rise in growth on DP, B and D media respectively. Pseudomonas sp. shows 8%, 48%, 33% rise in growth on DP, B and D respectively, which is higher than that of growth on commercial media. Alternative media could be used as cheap media for routine experiment in laboratory. On comparison with Nutrient Broth, our formulated media DP, D and B gave better results. Formulated media was found to be highly cost effectively.          Englishhttp://ijcrr.com/abstract.php?article_id=2421http://ijcrr.com/article_html.php?did=2421 A. Ravathie, P. Sevvel, R. Nirmala, N. Kularajany (2012). Journal of natural product and plant resources, 2; 697-700. Adesemoye AO and Adedire CO, Use of Cereals as basal medium for the formulation of alternative culture media for fungi, World J Microbiol and Biotech, 2005; 21: 329-336. Adoki A., (2008). Factors affecting yeast growth and protein yield production from orange, Platinum and banana waste processing residues using Candida sp. African. J. Biotechnology 7(3), 290-295. Anupama and Ravindra P. Value added Food: single cell protein (2000). Biotechnology advances,  18, 459- 479. Asad M. J., Asghan M., Yaqub M. and Shahzad K (2000) Production of single cell protein,. (2000). Delignified Corn Cob By Arachniotus species, Pak. J. Of Agric. Sci., 2000. 37, 3-4. Basu S, Bose C, Ojha N, et al. Evolution of bacterial and fungal growth media. Bioinformation. 2015;11(4):182-184. doi:10.6026/97320630011182 Bhattacharya S, Vijayalakshmi N, Parija S C. Uncultivable bacteria : Implications and recent trends towards identification. Indian J Med Microbiol 2002;20:174-7 Deivanayaki M and Iruthayaraj AP, Alternative vegetable nutrient source for microbial growth, Inter J Biosci, 2012; 2: 47-51. Famurewa and O.M. David, 2008. Formulation and Evaluation of Dehydrated Microbiological Media from Avocado Pear (Peasea americana Cmill). Research Journal of Microbiology, 3: 326-330.) Jamel P., Alam M. Z. and Umi N.,( 2008) Media optimization for bio proteins production from cheaper carbon source,.. J. of Engi. Sci. and Techno., 3(2) 124-130. Janssen PH, Yates PS, Grinton BE, Taylor PM, SaitM (2002) Improved culturability of soil bacteria and isolation in pure culture of novel members of the divisions Acidobacteria,Actinobacteria, Proteobacteria, and Verrucomicrobia.. Appl Environ Microbiol.;68:2391–6. Kasanadze AK (2000) Replacement of Agar by cassava flour in microbial media. BSc Dissertation University of Malawi, Bunda College of Agriculture, Lilongwe. Konstantinidis KT, Tiedje JM. (2005) Genomic insights that advance the species definition for prokaryotes..ProcNatlAcadSci USA;102(2567–2572):99. Kuria P, Demo P, Nyende AB, Kahangi EM. (2008) Cassava starch as an Alternative gelling agent for the in vitro micro propagation of potato (Solanum tuberosum L.).. African Journal of Biotechnology, .7: 301–307. Leach HW, McCowan LD, Schoch TJ. (1959) Swelling and solubility patterns of various starches..Cereal Chemistry. 36: 534-544. Lucyszyn, N., M.Quoirin, M. M. Homma and M. R. Sierakowski. (2008) Agar/galactomannan gels applied to shoot regeneration from tobacco leaves.. Biol. Plant. 51(1):173-176. Maliro MFA, Lameck G. (2004) Potential of cassava flour as a gelling agent in media for plant tissue cultures. African Journal of Biotechnology ,. 3: 244-274. Mbanaso ENA. (2008) Effect of multiple subcultures on Musa shoots derived from cassava starch-gelled multiplication medium during micropropagation. African Journal of Biotechnology Vol 7, No 24 (2008)   Mohamed, M. A. H., A. A. Alsadon and M. S. Al Mohaidib .( 2009) Corn and potato starch as an agar alternative for Solanumtuberosum micro propagation. .Afr. J. Biotechnol. 9(1): 012- 016. Murashige, T. and F.Skooga.( 1994) A revised medium for rapid growth and bio assays with the tobacco tissue cultures.. Physiol. Plant. 15: 473-497. N. Ravimannan, R. Arulanantham, S. Pathmanathan and K. Niranjan. (2014) Annals of Biological Research,.    5(1), 36-39. Naik, P. S. and D. Sarker. ( 2001) Sago An Alternative Gelling Agent For Potato In Vitro Culture..Biol.Plant.  44(2): 293. Najafpur, Ghasem D. (2007) Single Cell Protein. Biotechnology advances. Biochemical Engineering and Biotechnology Advances, .332-347. Nambiar VS, Bhdalkar k, Daxini. (2003) Drumstick Leaves As Source Of Vitamin A In ICDS-SFP.. Indian Journal of Pediatrics.70: 383-387.  Nambiar VS, Mehta R, Daniel M. (2005) Polyphenol Content of Three Indian Green Leafy Vegetables.2005. Journal of  Food science and Technology.,42: 312-315. Nambiar VS, Seshadri S. (1998.) Beta Carotene Cotent Of Green Leafy Vegetables Of Western India By HPLC. Journal of Food Science and Technology. 35: 365-367.  Nichols D. (2007) Cultivation gives context to the microbial ecologist..FEMS Microbiol E.coli.; 60:351–7. Ozel, C. A., K. M. Khawar and O. Arslan  (2008).Comparison Of The Gelling Of Isubgol, Agar And Gelrite  On In Vitro Shoot Regeneration And Rooting Of Variety Samsun Of Tobacco (Nicotiana tabacuml.).. Scient.Horticul.2008. 117(2): 174-181. Pandhre GR, Satwase AN, Hashmi  SI. (2011) Effect Of Different Pretreatments And Drying Methods On Color Characteristics Fenugreek Leaves., Int J Cur Sci Res 1: 115-119. Pham VHT, Kim J. Cultivation Of Unculturable Soil Bacteria.(2012). Trends Biotechnology . 30:475–84. Porndarun, H. Vichai. and C.Walairut. (2010) Department of product development, Faculty of agro industry , Kasetsart University, Bangkok 10900,Thailand,. 100-106. Priadi, D., H. Fitriani and E. Sudarmonowati. (2008) The growth  of cassava (Manihotesculenta Crantz)on various alternative gelling agents. Biodiversitas 9(1): 9-12. Prokash, S., M. I. Hoque and T. Brinks.( 2000) Culture media And Containers,. Biotechnology And Eco Development Research Foundation, Bangalore, India.. S. Tharmila, E. C. Jayaseelan and A. C. Thavaranjith (2011) Archives of applied science Research,.3; 389-393. S. Tharmila, E. C.Jeyaseelan and A. C. Thavaranjit (2011) Archives of Applied Science Research. Vol.3 (3):389-393. Sathiya Vimal S, Vasantha Raj. S , Senthil kumar. R. P , and Jagannathan. (2013) Natural Sources of Gooseberry Component used for Microbial Culture Medium Journal of Applied Pharmaceutical Science Vol. 3 (11), 040-044. Vartoukian SR, Palmer RM, Wade WG. (2010) Strategies for culture of ‘unculturable’ bacteria. FEMS Microbiol Lett. 309:1–7. Zapata, A. Cost Reduction In Tissue Culture Of Banana.(2001). (Special Leaflet), International Atom Energy  Agency Labs., Agric. And Biotech. Lab., Vienna, Austria. Zhe, W. R., C. Debasis, E. J. Hahn and K. Y. Paek. (2005) Growth Of Doritaenopsis In Peat substituted Growing Medium. J. Korean Soc. Hort. Sci.. 46(1): 76-81. Wasas AD, Huebner RE, Klugman KP. Use of Dorset Egg Medium for Maintenance and Transport of Neisseria meningitidis and Haemophilus influenzae Type b. Journal of Clinical Microbiology. 1999;37(6):2045-2046.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241102EnglishN2018January19HealthcareComparative Evaluation of the Antibacterial Activity of Calcium Hydroxide in Combination with 2% Chlorhexidine and Ozonated Chlorhexidine Against E.faecalis – An Invitro Study English1618Ravivarman C.English Karthikeyan A.English Senthilkumaran M.English Josephin Shalini R.EnglishIntroduction: Calcium hydroxide has been used with various vehicles in the treatment modalities of Endodontics. Owing to the increased antibacterial resistance, Ozone is currently being discussed as a possible alternative vehicle because of its reported high antimicrobial property without the development of drug resistance. Aim: The purpose of this study was to investigate and compare the effectiveness of calcium Hydroxide in combination with 2% chlorhexidine and ozonated chlorhexidine for elimination of E.faecalis bacteria. Materials and Methods: In this study aqueous ozone was used, it was prepared by passing gaseous ozone through CHX in a glass beaker. Minimal inhibitory concentration of ozonated CHX was obtained by serial dilution method. The samples were assigned to 3 groups: Group I- Calcium hydroxide with saline; Group II- Calcium hydroxide with 2% Chlorhexidine; Group IIICalcium hydroxide with ozonated chlorhexidine. Efficacy of intracanal medicaments against E.faecalis was carried out by disc diffusion method. Results were statistically analysed by one way ANOVA. Results: The antimicrobial effectiveness of the samples in Group II where ozone was ozonated with reduced percentage i.e. 0.12% was around 21mm which is almost equal to that of 2% CHX and the results were found to be statistically significant at 0.5. Conclusion: Ozonated CHX with reduced concentration possessed similar antimicrobial effect as conventional CHX in combination with calcium hydroxide. EnglishOzone, Chlorhexidine, Calcium hydroxide, Zone of inhibitionIntroduction: The success of root canal treatment relieson biomechanical preparation, irrigation, microbial control and complete obturation of the root canal system. Bacteria and their products are considered the primary etiologic agents of pulp necrosis and peri-radicular lesions1. Due to anatomical complexities and consequent limitations of access by instruments and irrigants, microorganisms infecting the root canal may survive endodontic procedures. Microorganisms may also infiltrate through a poor temporary seal during the period betweenappointments for endodontic treatment. Therefore, the use of a root canal dressing is important for obtaining and maintaining a disinfected canal after mechanical instrumentation and before root canal obturation.  Despite of its viscous consistency, the paste made by mixing calcium hydroxide and chlorhexidine (CHX) gel 2% was found not to influence the sealing ability of the obturation technique. Calcium hydroxide plays an important role in endodontics due to its ability to induce hard tissue formation, its antibacterial effectand its ability to act as a physical barrier toprevent root canal reinfection2.   In  an  attempt  to  enhance  the  anti-microbial activity of calcium hydroxide, different substances have  been  used  as  vehicles.  Chlorhexidine glucometer has been widely used as an endodontic irrigant, because of its wide antimicrobial activity against Gram-positive and Gram-negative microorganisms. CHX may also present residual antimicrobial activity on the dentin surface after prolonged with the root canal3. Studies have suggested that CHX could be used in combination with calcium hydroxideto improveantimicrobial efficacy against resistant microorganisms. However a meta- analysis conducted by Mastoid Saatchi concluded that mixing calcium hydroxide with CHX does not improve antibacterial property as intracanal medicament against E.feacalis. Currently, ozone has beendiscussed as a possible alternative antiseptic agent in dentistry because of its reported high antimicrobial power without the development of drug resistance. Ozone gas in a concentration of 4g/m3(Heal Ozone; Kavo, Liberace, Germany) is already being used clinically for endodontic treatment4. Regarding the demand on relative non-toxicity toward periapical and oral mucosal tissue  for  the  endodontic  irrigants,  the  ozone  gas  concentration currently used in endodontics (4 g/ m3) has been shown to be slightly less cytotoxic than NaOCl (2.5%) and aqueous ozone (up to 20 μg/mL) showed essentially no toxicity to oral cells in vitro. Hence the purpose of the study is to compare the antibacterial effectiveness ozonated CHX in combination with Calcium hydroxide and that of calcium hydroxide in combination with 2%CHX. Materials and methods: Preparation of samples: In this study aqueous ozone was used, it was prepared by passing gaseous ozone through CHX in a glass beaker. As a preliminary step, the minimal inhibitory concentration of ozonated CHX was obtained by serial dilution method. The samples were assigned to 3 groups: Group I- Calcium hydroxide was mixed with saline; Group II- Calcium hydroxide was mixed with 2% Chlorhexidine; Group III- Calcium hydroxide was mixed with ozonated chlorhexidine. Isolation of micro-organisms: Pure strain of Enterococcus faecalis from American type culture (ATCC #29212) was used. Respectively cultures were grown over-night at 37°C in brain heart infusion (BHI) broth on a rotary shaker 150 rpm and microbial growth were checked by changes in turbidity at 24 hours. Antimicrobial activity testing: Antimicrobial efficacy of the assigned samples wastested against E.faecalis using agar disc diffusion method. Mueller-Hinton broth was selected and the plates were incubated at 37° C in bacteriological incubator for 48h. Wells of diameter 6mm and depth 3mm were made and the samples were subjected to antimicrobial testing. Zone of inhibition: Samples were placed in the prepared wells and thediameter of inhibition areas of microbial growth was measured after 48h with a millimetrecalliper, for each sample, which corresponded to the substance’s antimicrobial potential. Results: The antimicrobial effectiveness of the samples in Group II where ozone was ozonated with reduced percentage i.e. 0.12% was around 21mm which is almost equal to that of 2% CHX and the results were found to be statistically significant at 0.5. Discussion: The success of root canal treatment lies in the complete elimination of microorganisms. E.faecalis is the most common microorganism present in the persistent per-radicular infection. Due to its versatile nature various medicaments have been tried to completely eliminate from root canals. And also owing to the increased antimicrobial resistance, alternatives to antibiotics have been tried5. Ozone, due to its interesting biologic properties can be used to treat root canal infection with reduced antimicrobial resistance and side effects6. Ozonated oils with zinc oxide eugenol when used as root filling material in primary tooth had shown good success rate7. Chlorhexidine has been proved to be effective against E.faecalis and to improve its properties ozone can be incorporated.at the same time, a meta-analysis concluded that mixing of CHX with calcium hydroxidehas no additive effect8. In this study, ozonated chlorhexidine has been used in combination with calcium hydroxidefor elimination of E.faecalis. The samples were assigned to three groups and subjected to disc diffusion method for antimicrobial efficacy. After 48 hours the antimicrobial efficacy was evaluated by measuring the zone of inhibition. The antimicrobial efficacy of 0.12% ozonated CHX in combination with calcium hydroxidewas almost equal to that of  2% CHX with calcium hydroxide. Conclusion:Since the cytotoxic effects of CHX has been found to be dose dependant, ozonated CHX with reduced percentage and with added effects of ozone seems to be a better alternative. However further clinical trials and randomized control studies have to be done to use it as an intracanal medicament invivo. Englishhttp://ijcrr.com/abstract.php?article_id=2422http://ijcrr.com/article_html.php?did=2422 C. M. Sedgley, S. L. Lennan , O. K. Appelbe  Survival of Enterococcus faecalis in root canals ex vivo. International Endodontic Journal. 2005; 38:735-42. Zerella JA,  Fouad AF, Spangberg LSW, Effectiveness of a calcium hydroxide and chlorhexidine digluconate mixture as disinfectant during retreatment of failed endodontic cases. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2005;100:756-61. Zamany A, Safavi K, Spangberg LS. The effect of chlorhexidine as an endodontic disinfectant. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2003;96(5):578-81. Baysan A, Lynch E. Effect of ozone on the oral microbiota and clinical severity of primary root caries. Am J Dent. 2004;17(1):56– 60.1  Nagayoshi M, Kitamura C, Fukuizumi T, Nishihara T, Terashita M. Antimicrobial effect of ozonated water on bacteria invading dentinal tubules. J Endod. 2004;30(11):778–81.8.  Estrela C, Estrela CR, Decurcio DA, Hollanda AC, Silva JA. Antimicrobial efficacy of ozonated water, gaseous ozone, sodium hypochlorite and chlorhexidine in infected human root canals. Int Endod J. 2007;40(2):85–93. Chandra SP, et al Success of root fillings with zinc oxide-ozonated oil in primary molars:  preliminary  results. Eur Arch Pediatr Dent 2014;15;191-5 Masoud Saatchi, et al. antibacterial effect of calcium hydroxide combined with chlorhexidine on Enterococcus faecalis: a systematic review and meta-analysis. J Appl Oral Sci. 201;22(5):356-65.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241102EnglishN2018January19HealthcareA Case of Congenitally Corrected Transposition of the Great Arteries (CC-TGA) Associated with Intracardiac Anomalies with Complete Situs Inversus and Dextrocardia English1921Ankur MittalEnglish Rattilal MeenaEnglish Sudheer MaluEnglish Neera SamarEnglishA case of congenitally corrected transposition of the great arteries (CC-TGA) associated with intracardiac anomalies – pulmonary stenosis, Ventricular septal defect (VSD), supravalvular Aortic stenosis (AS), with complete situs inversus and dextrocardia. The CC-TGA is rare but its association with intracardiac anomalies and rhythm disorders is not rare. EnglishCongenital heart disease, Transposition of great arteries (TGA), Ventricular septal defect (VSD), Situs inversusINTRODUCTION  CC-TGA describes heart in which there are discordant atrioventricular connections in combination with discordant ventriculoarterial connections (double discordance). This is a rare complex adult Congenital heart disease (CHD) ( 75%), congenital complete heart block (CHB) (5%) 2 and dextrocardia.4           The mirror image dextrocardia is usually observed with complete situs inversus, which occurs most frequently in individuals whose hearts are otherwise normal. In this article we report a 18 year male patient presented with CC-TGA, pulmonary stenosis, VSD, supravalvular AS, with complete situs inversus and dextrocardia.     CASE HISTORY                                                                              A 18 year old boy admitted in our hospital with dyspnea on exertion off and on since 3 years, fever with chills off and on 4 days, jaundice of 2 days duration. No history of palpitation, syncope, chest pain and swelling feet. Clinically jaundice was present. Cyanosis, clubbing, edema was absent, jugular venous pressure was normal, Blood Pressure -110/70mmhg, pulse-normal, and no signs of  Infective endocarditis seen . Liver was mildly enlarged and palpable in left hypocondrium and spleen mildly palpable on right hypocondrium. Chest shape was normal. Cardiac apex beat present in right 5th Intercoastal space - 4cm from midsternal line. First heart sound was unremarkable. Second heart sound was single and soft. No third and fourth heart sound.  Harsh systolic murmur grade3/6 heard at Right sternal border in 3rd-4th Intercoastal space. Auscultation of lungs was normal. On investigations: Hemoglobinb12gm%, Peripheral blood film shows P. vivax ring forms, S. bilirubin: 2.5 mg% with direct 0.3, indirect 2.2; Liver transaminases were normal. Chest X-ray posterioanterior view was suggestive of mesocardia, and gastric bubble seen below right dome of diaphragm. Electrocardiography suggestive of dextrocardia, inversus position of atria. 2D Echocardiography & doppler study suggestive of dextrocardia, CC- TGA, Pulmonary stenosis (PS), VSD, supravalvular AS, no Pulmonary artery hypertension, no vegetations. CT thorax-abdomen suggestive of  situs inversus with right sided lung showing two lobes and left sided lung showing three lobes. Blood for culture (3sets), separated from one another by at least 1hr, obtained from different venipuncture sites were negative after 72hours. Our clinical diagnosis was malaria fever, congenital Heart disease – CC-TGA, dextrocardia, complete situs inversus, pulmonary stenosis, VSD, supravalvular AS.  DISCUSSION In CC-TGA there is usual atrial arrangement, systemic venous blood passes from the right atrium through a mitral valve to a left ventricle and then to the posteriorly located pulmonary artery. Pulmonary venous blood passes from the left atrium through a tricuspid valve to a left sided right ventricle and then to an anterior, left-sided aorta. Circulation is thus functionally (physiologically) corrected but the morphological right ventricle supports the systemic circulation.           The associated intracardiac anomalies in our presented case are PS, VSD, and supravalvular AS. Heart blocks, arrhythmias not present.           Patients without associated abnormalities (isolated CC-TGA) can exceptionally survive until 7th or 8th decade.1 Progressive systemic (tricuspid) Atrioventricular valve regurgitation and system (right) ventricular dysfunction tend to occur from 4th decade onward, whereas atrial tachyarrhythmias are more common from 5th decade onward.3 Patients with associated anomalies (pulmonary stenosis, left-sided tricuspid valve anomaly or VSD) often require surgical palliation (systemic-to-pulmonary artery shunt for cyanosis) or repair of associated anomalies i.e., tricuspid valve replacement and double-switch procedure.           Adult patient with CC-TGA may develop Congestive heart failure with exertional dyspnoea, palpitation, syncope secondary to complete Heart block or cyanosis (if Left ventricular outflow tract obstruction and VSD are present). Clinical features depend on presence of associated lesions and complications.   Our patient presented with breathlessness on exertion, without palpitation, cyanosis, edema clubbing, and syncope.         This case was treated empirically, and advised surgical correction of intracardiac anomalies at higher centre, but patient’s relatives refused because they did not afford costly treatment.           Our case is interesting, though not uncommon in pediatric cardiac practice, this case has a potential to create awareness for general practitioners also. As our best efforts, we search in literature, few such cases reported in literature till date. Conclusion Transposition of the Great Arteries (CC-TGA) Associated with Intracardiac Anomalies with Complete Situs Inversus and Dextrocardia in adult is interesting, though not uncommon in pediatric cardiac practice, this case has a potential to create awareness for general practitioners also. Patients with corrected TGA represent an important group of adults with congenital heart disease. Most patients are asymptomatic but require ongoing, knowledgeable follow-up by specialized care-givers to optimize their longevity and quality of life. Acknowledgment Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Englishhttp://ijcrr.com/abstract.php?article_id=2423http://ijcrr.com/article_html.php?did=2423 Roffi M, de Marchi SF, Seiler C: Congenitally corrected transposition of the great arteries in an 80 year old woman. Heart 79:622-623, 1998. Webb G et al.: Congenital heart disease. In Braunwald’s Heart Diseases: A Text Book of Cardiovascular Medicine, 7th ed, Zipes D et al. (eds), Philadelphia, Elsevier Saunders, 2005. Kafali G, Elsharshari H, Ozer S et al.: Incidence of dysarrhythmias in congenitally corrected transposition of the great arteries. Turk J Pediatr 2002; 44:219-223. Elisabeth Bedard, Michael A Gatzoulis: Adult congenital heart disease, evidence-based cardiology, 3rd edition, by S. Yusuf, J.A. Cairns, ©2010 Blackwell publishing, 1020-21.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241102EnglishN2018January19HealthcareA Rare Case Report of Hydrometrocolpos in a Female Newborn English2224Khadija KansoEnglish Bassem Abou MerhiEnglish Marwan ZeidanEnglish Soukaina IbrahimEnglish Fatima GhandourEnglish Fadi IskandaraniEnglish Eliane El-HouwayekEnglish Fatima YassineEnglish Imad ChokrEnglishIntroduction: Congenital hydrocolpos and the hydrometrocolpos is a rare disorder caused by accumulation of cervico-vaginal secretions that present with cystic pelvic mass in neonates.Prenatal diagnosis is important to prevent complications like sepsis and renal failure and to affect prognosis. We will report a case of a newborn female admitted to our NICU with abdominal mass and imperforated anus diagnosed to have hydrometrocolpos, septated vagina with bicornate uterus and urogenital sinustreated surgically. Case Report: This report will help us to develop a high index of suspicion of hydrometocolpos in female newborn with abdominal mass. EnglishHydrocolpos, Hydrometrocolpos, Neonatal Intensive Care Unit (NICU).Introduction: Hydrocolpos and hydrometrocolpos is uncommon disorder caused by vaginal distention and fluid accumulation (1). Congenital hydrocolpos can be associated with other genitourinary anomalies or it can be a part of a syndrome. Prenatal diagnosis either by ultrasound or magnetic resonance imaging is essential to prevent complications (1, 2). Case Report: We report a case of 35weeks and 5 days preterm girl with history of normal vaginal delivery to a 19 years old mother, G1P1A0 admitted to our neonatal intensive care unit (NICU) with abdominal distension, lower limbs swelling and imperforated anus. The pregnancy though uncomplicated was poorly followed and no antenatal ultra-sonograms (US) were available. On physical examination, the baby had a palpable mass in the lower mid abdomen. The superior margins of the mass were well felt and appreciated. Both lower limbs found to have pitting pedal edema. She had low imperforated anus. The child was referred to Department of Radio- diagnosis for evaluation of the abdominal mass. A plain radiograph of the abdomen (KUB) was initially obtained and showed homogenous soft tissue opacity in the suprapubic region of the abdomen that was causing peripheral displacement of the bowel loops(Figure 1).   Ultrasound of the abdomen was done at the same time and showed that there was a huge fluid filled structure occupying almost all the abdomen and showing a midline septations. This mass was causing displacement of the liver superiorly. Bowel loops were being displaced peripherally. There was also bilateral hydronephrosis. Rest of the intra- abdominal organs appeared to be normal (Figure 2). Following ultrasound, a preliminary diagnosis of neonatal hydrometrocolpos was made. A computed tomography scan (CT Scan) of the abdomen and pelvis was done immediately after KUB and US to reveal a cystic mass 8.5x8.2x6 cm occupying the abdomen and pelvic cavity with median septations and double thin walls extending down to the rectum with severe bilateral hydronephosis. The uterus was not well dilneated (Figures 3a, 3b, and 3c).   The child was transferred to the operating room (OR) where imperforated hymen,septated vagina with bicornate uterus, hydrometocolpos and urogenital sinus (vagina was connected to the urethra) were identified. Drainage of milky intravaginal collection and closure of the communication between urethra and vagina was performed. Foley catheter was inserted through the cloaqual canal into the vagina for continuous drainage and lastly colostomy was performed. After stabilization in the NICU the patient was discharged from our institute to be followed up later for more exploration of the ano-genital area firstly for anatomical definition and secondly for surgical anastomosis and closure of the colostomy after 6 months. Discussion and Review of Literature: Hydrocolpos is an uncommon disorder that can be associated with genitourinary anomalies ranging from persistent urogenital sinus to cloacal dysgenesis (1). It can be defined as vaginal distention with fluid accumulation and it can be due to either a combination of increased activity of secretory cervical glands and vaginal obstruction or the presence of urogenital sinus with urine collection (1, 2). Hydometrocolpos is characterized by uterine distention that might result if the accumulated fluids stretchthe cervical canal as well as the body of the uterus (2). The causes of accumulation of cervico-vaginal secretions are diverse and include imperforate hymen which is defined as a membrane occluding the lower third of vagina, transverse vaginal septum, vaginal atresia and malformations of cloaca including urogenital sinus (2,3). Cloacal malformations form a group of non-hereditary ano-rectal malformations. The cloaca is defined as a single common chamber of the caudal intestinal, urinary, and genital tract; it is normally present in the 4th to 5th week embryogenesis. Later in the fetal life the cloaca will be divided into the urogenital sinus anteriorly and ano-rectum posteriorly. The urogenital sinus then becomes the urinary bladder and urethra with a portion transformed into the vagina and hymen. The type of cloacal anomalies will results from the timing of developmental arrest (4). Hydrometrocolposis rare with an incidence of 0.006% approximately (2, 3). Bischoff et al. reported a high incidence of hydrometrocolpos in patients with cloacal malformations (28%) (5). Child with this disorder presents clinically with increasing abdominal distension (3). Prenatal diagnosis of hydrometrocolpos by radio-imaging may be difficult but is essential in that it can substantially affect prenatal and immediate neonatal treatment and it can help improving prognosis (4).Ultrasonography and in particular magnetic resonance imaging can be helpful in the antenatal diagnosis of congenital hydrometrocolpos. Magnetic resonance imaging (MRI) provides informationin cases in which the diagnosis is unclear or additional anomalies cannot be adequately evaluated by ultrasound. MRIproved to be better when ultrasonography is limited because of late presentation or by the presence of obesity or oligohydramnios (3, 4). Antenatal imaging will be helpful for preoperative planning (4) and for appropriateprenatalcounseling to parents. It may improve the outcomes especially related to obstructive uropathies(1). Association of an imperforate hymen with other genitourinary anomalies is well known such as imperforate anus, cloacal anomalies, renal agenesis, uterine anomalies or persistent urogenital sinus. It may be a part of a syndrome like McKusick-Kaufman syndrome (autosomal recessive disorder which includes vaginal atresia and secondary hydrometrocolpos, polydactyly, congenital cardiac anomalies and hydrops fetalis), Ellis Van Creveld syndrome, Bardet- Biedl syndrome, VACTERL association (Vertebral, anal, cardiovascular, tracheooesophageal, renal and limb anomalies), and MURCS association (Mullerian duct aplasia/hypoplasia, renal agenesis/ectopia and cervico-thoracic somite dysgenesis such as Klippel Feil abnormality, anomalous ribs or Sprengel deformity) (1, 2, and 3).The most frequent complication of congenital hydrometrocolpos includesurinary tract obstruction, leading to hydronephrosis and renal failure. Other complications include repeated urinary tract infections, sepsis, pyocolpos, rupture and peritonitis (1, 2, and 3). Drainage of the collection which can be performed under ultrasound guidance was associated with decreased risk of urinary obstruction and associated complications (3, 4). The recognition of hydrocolpos prenatally can facilitate rapid treatment at birth (4). Infants with prenatal diagnosis of pelvic mass should be categorized as high risk deliveries and should be promptly followed by multidisciplinary team soon after birth (1). Conclusion: Hydrocolpos or hydrometrocolpos is uncommon disorder of neonates that can present with a midline abdomino-pelvic mass. Ultrasound and magnetic resonance imaging can help in the prenatal diagnosis and early surgical treatment for better outcome and prognosis. This report will aid us to the development of high index of suspicion of congenital hydrocolpos in neonates with pelvic mass for early and even antenatal diagnosis that lead to further prevention of secondary complications such as hydronephrosis and gastrointestinal obstruction. Acknowledgement: Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Englishhttp://ijcrr.com/abstract.php?article_id=2424http://ijcrr.com/article_html.php?did=2424 Murthy V, Costalez J, Weiner J, Voos K. Two Neonates with Congenital Hydrocolpos. Case Reports in Pediatrics, vol. 2013, Article ID 692504, 3 pages, 2013. Shetty D, Varma R. Neonatal Hydrocolpos: A Case Report and Review of Literature. SSRG International Journal of Medical Science. 2017;4. B R, Basavalingu D, Paramesh VM, NagendraPDK. Radiological Diagnosis of Neonatal Hydrometrocolpos- A Case Report. J Clin Diagn Res. 2016; 10(3): TD18–TD19. Winkler N,Kennedy A, Woodward PJ. Cloacal Malformation. J Ultrasound Med. 2012; 31:1843–1855. Bischoff A, Levitt MA, Breech L, Louden E, Pe˜na A. Hydrocolpos in cloacal malformations. Journal of Paediatric Surgery. 2010;45:1241–45.

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241102EnglishN2018January19HealthcareIn Vitro Interaction Studies between Artemether – Lumefantrine and Lamivudine/Metronidazole English2530Awofisayo Sunday O.English Arhewoh Matthew I.English Okhamafe Augustine O.EnglishThis work assesses tablets of artemether and lumefantrine in vitro drug interaction with lamivudine or metronidazole. Spectra changes on artemether or lumefantrine vibration bands were evaluated using Fourier transform infrared spectroscopy (FTIR) and analyzed with essential FTIR (eFTIR) software. Instantaneous pH changes and acid buffering capacity in biorelevant media were also determined. USP type-2 dissolution apparatus (paddle) containing Fed State Simulated Intestinal Fluid Version 2 (FeSSIF-V2) was employed for dissolution test. Sample (5 mL) collected at various predetermined time intervals were analyzed simultaneously for artemether and lumefantrine with high performance liquid chromatographic (HPLC) reverse phase (RP) system, at 25oC. Artemether (O-H) stretching vibration was shifted to wavenumber 3387.0 and 3408.22 cm-1 by lamivudine and metronidazole, respectively. There was no significant shift in spectral bands corresponding to the endoperoxide linkage due to both drugs. Lamivudine and metronidazole showed no significant change in the pH of biorelevant media (p > 0.05). The release kinetics in FeSSIF-V2 for artemether changed from Higuchi (R2 = 0.9124) to first order (R2 = 0.9422) due to presence of lamivudine while that of lumefantrine from first order (R2 = 0.9423) to Higuchi due to the metronidazole (R2 = 0.9871). There was no significant level of interaction between lamivudine and metronidazole with the actives of AL tablet in vitro. The drugs therefore can be co-administered without any biopharmaceutical implications. EnglishRelease kinetics, Artemether-lumefantrine, Lamivudine, MetronidazoleINTRODUCTION Co-prescribing and use of more than one drug at the same time is regular occurrence in clinical practice for the treatment of a single or multiple pathological conditions (Bennett and Brown, 2008). The concurrent use of multiple medicinal agents has been further necessitated by co-morbidities of pathologic conditions such as HIV/AIDS, diabetes, hypertension and malaria infection (Ahsan et al., 2012). Malaria is one of the widely reported infectious diseases in the world that caused about one million deaths in the year 2006 (Rosenthal, 2014). The artemisinin derivatives have been co-formulated with other antimalarial agents and used with success in areas earlier reported with cases of multi-drug resistant Plasmodium falciparum infection (Adjei et al., 2008, Sagara et al., 2001). Some diseases of global interest along with malaria include intestinal parasitic infections such as amoebiasis or giardiasis and HIV/AIDS (Haque et al., 2003; CDC 2008, Cohen et al., 2008). Artemether-lumefantrine (AL) is a fixed dose combination (FDC) antimalarial that is widely prescribed based on World Health Organization recommendation for uncomplicated Plasmodium falciparum malaria (WHO, 2015). This drug however may be co-prescribed with other drugs for which malaria has co-morbidities or share geographical distribution. In the management of human immunodeficiency virus (HIV) infection, most clinicians prescribe fixed dose combinations of either tenofovir/emtricitabine or abacavir/lamivudine both of which can be given once daily. Zidovudine/lamivudine is commonly reserved for second or third line regimen due to toxicity and dosing schedule (Schooley, 2010; Thompson et al., 2010). Lamivudine as antiretroviral drug (ARD) and for hepatitis or metronidazole for intestinal and hepatic forms of amoebiasis/giardiasis may be co-administered with antimalarial AL. Antimalarial drug resistance has been of global concern with searchlight directed at avenues that predicate the bioavailability of the drugs. Drug - drug interactions (DDI) have been reported as consequent to treatment failure (Edwards and Aronson, 2000). A study on drug-drug interaction of pyronaridine/artesunate and ritonavir in healthy volunteers by Morris and co-workers concluded that co-administration of ritonavir and pyronaridine/artesunate interacts to alter exposure to artesunate, dihydroartemisinin and ritonavir itself (Morris et al., 2012). Several in vitro models have been developed for predicting in vivo drug interactions of co-administered dugs (Wienkers and Heath, 2005).   Fourier transform infra-red spectroscopy (FTIR) utilizes the phenomenon of wave interference and a software Fourier transform of the interferogram, to produce an infra-red spectrum that is characteristic of molecules. This instrumental method has been exploited extensively as an in vitro approach to predicting possible in vivo DDI (Kumari and Balaji, 2013). This study seeks to evaluate in vitro drug–drug interaction between AL and lamivudine or metronidazole. MATERIALS AND METHOD Materials AL tablet (Coartem®) was bought from a registered drug outlet, in Uyo State, Southern Nigeria. Lamivudine and metronidazole tablets were purchased in Lagos State, Nigeria. The details of the drugs are presented in Table 1. Acetonitrile, methanol, tetrahydrofuran (THF), potassium dihydrogen phosphate were HPLC grade, products of Sigma Aldrich, Germany. FaSSIF/FeSSIF/FaSSGF and FeSSIF-V2 powder are products of Biorelevant.com, UK.  Other reagents were of analytical grade, products of Sigma Aldrich, Germany. Methods Preparation of Standard solutions The internal standard (IS) solution was prepared by accurately weighing 10 mg of halofantrine into 10 mL volumetric flask. A volume of 6 mL of methanol was added to dissolve and subsequently made to mark to produce a stock concentration of 1 mg/mL. A weight of 20mg and 120mg of reference standard artemether and lumefantrine, respectively were poured into different 10 mL volumetric flask. Artemether and lumefantrine were dissolved in acetonitrile and tetrahydrofuran (THF), to produce 2 mg/mL and 12 mg/mL stock solutions, respectively. Mixed standard solutions of artemether – lumefantrine and (IS) were prepared by adding equal aliquot volume of artemether and lumefantrine stock solutions (2 mL), dispensed into 5mL plain polypropylene sample bottles to produce 1 mg/mL and 6 mg/mL of the standards, respectively. Serial dilutions of the mixed standards and reference standard solutions were made to obtain graded concentrations of 0.01/10, 0.1/10.0, 0.5/20.0, 1.0/40.0, 5.0/60.0, 10.0/80.0 and 20.0/100.0 mg/mL.  The mixed standard solutions were diluted with acetonitrile and THF (50:50 %, v/v). The obtained solutions were spiked with IS stock solution to give 5 µg/mL with micropipette (Huang et al., 2010).   Preparation of buffers and simulated intestinal solution A weight of 1.0 g of sodium chloride was dissolved in 0.450 L of distilled water. The pH of the resulting solution was adjusted to 1.6 with hydrochloric acid solution and made up to 0.5 L with distilled water at room temperature. A weight of 2.020, 4.325 and 5.937 g of sodium hydroxide pellet, glacial acetic acid and sodium chloride, respectively, were dissolved in 0.450 L of distilled water and the resulting pH adjusted to 5.0 with either of 1N sodium hydroxide or hydrochloric acid. Fasted state simulated gastric fluid (FaSSGF) was prepared by dissolving 1.120 g of FaSSIF/FeSSIF/FaSSGF powder in 0.25 L of pH 1.6 buffer. This was stirred until the powder was completely dissolved. The solution was made up to 0.5L mark. The solution was allowed to stand for 2 hours before use and an unused portion was discarded after 48 h of preparation. Fed state simulated intestinal fluid was prepared by dissolving 5.60 g of FaSSIF/FeSSIF/FaSSGF powder in 0.25 L of buffer pH 5.0. The solution was made up to 0.5 L volume and allowed to stand for 2 h before use. Any unused solution was discarded after 48 h.Fed State Simulated Intestinal Fluid – Version 2 (FeSSIF – V2) was prepared by dissolving 5.0 g of FeSSIF – V2 powder in 5 L of distilled water (Galia et al., 1998). Assessment of tablet quality parameters The weight uniformity, tablet friability, disintegration and hardness were performed based on established pharmacopeia protocols. The chemical content of artemether and lumefantrine were simultaneously determined using the chromatographic system. FTIR Spectroscopic analysis FTIR spectroscopy was performed using FTIR spectrophotometer (FTIR 84005,Schimadzu, Japan). One milligram of AL crushed powder and 200 mg of dried potassium bromide (KBr) powder were mixed in a mortar and compressed into a translucent disk. The scanning was carried out at a speed of 2 mm/s over a wavenumber region of   4000 to 500 cm-1. Similarly, the process was repeated with the addition of 1 mg of the co-administered drugs (i.e, lamivudine or metronidazole) to the AL before pellet formation with KBr. The resulting spectra were analyzed using essential FTIR (eFTIR) software. Buffering capacity of drugs A tablet of lamivudine (150 mg) or metronidazole (200 mg) was crushed and dispersed in 500 mL of FaSSGF and FeSSIF separately. The change in pH of the medium was observed as the instantaneous pH change due to the added drugs. The amount of hydrochloric acid that produced one unit pH change with continuous pH monitoring was also determined. Dissolution studies A tablet each of lamivudine or metronidazole was dissolved in 500 mL of FeSSIF-V2 in a dissolution apparatus (USP type 2, paddle method). A tablet of AL was placed in the apparatus with speed of agitation and temperature at 100 rpm and 37oC, respectively. A volume of 5 mL of the dissolution media was sampled at 5, 15, 30, 45, 60 and 90 min. The sampled volume was filtered using 0.45 µm syringe filter. The filtrate was analyzed for artemether and lumefantrine concentration simultaneously using HPLC system. Statistical analysis All experiments were performed in triplicates and the differences in the effects of lamivudine or metronidazole on artemether and lumefantrine profile were analyzed statistically using single factor analysis of variance and statistical significant difference was taken at p < 0.05. RESULTS AND DISCUSSION In this study, FeSSIF-V2 was employed for AL dissolution studies as this medium has been extensively used to predict in vivo drug release (Lakka and Goswami, 2012). The content of FeSSIF-V2 such as sodium taurocholate and lecithin makes the output of this biorelevant medium to closely mimic intestinal environment (Vertzoni et al., 2005).The use of this updated postprandial biorelevant media, FeSSIF-V2, presents a readily available dissolution testing tool for quick adjudging of in vivo performance of drugs. In the model simulating the co-administration of lamivudine or metronidazole with AL, the result of instantaneous pH change due to lamivudine or metronidazole on FeSSIF produced a pH increase of 0.13 and 0.17 units, respectively (Table 2). The increases were however not significant as the pH were still within the range of physiological value for intestinal absorption of drugs. In the stomach-simulated media (FaSSGF), the instantaneous pH change result gave an increase of 0.31 and 0.55 units, respectively. Since a pH change of less than 1 unit was recorded in the gastrum, the ratio of unionized to ionized form of the dissolved drugs (i.e., artemether and lumefantrine) becomes insignificant.   The R2 value for the determination of artemether and lumefantrine were 0.9920 and 0.9930, respectively. The dissolution curve for artemether and lumefantrine are presented in Figure 1a and 1b, respectively. It was observed that artemether had significantly higher drug release in the presence of lamivudine (p < 0.05).but there was no significant difference in the drug release profile for lumefantrine in the presence of either lamivudine or metronidazole. An ideal formulation releases the exact amount of the active ingredient at the right time thereby optimizing the therapeutic action of the active ingredient (Dey et al, 2012). Drug release and other effects of co-administration of AL with other drugs have not been extensively evaluated. The dissolution profile for artemether and lumefantrine are expressed in Table 3. The C40 value for artemether in the medium containing metronidazole was significantly lower than that with lamivudine (25 versus 65 mg; p < 0.05). Media with metronidazole did not achieve 70% artemether release throughout the dissolution period. There was no difference in the dissolution profile for lumefantrine with respect to the additives (9.0 versus 10.0 mg). Lumefantrine did not achieve 70% release for the additives throughout the dissolution period. The R2 values of the media conditions describing the kinetic model that best describes the dissolution curves for artemether and lumefantrine drug release from  AL tablet were Higuchi (R2 = 0.9124) and first-order (R2 = 0.9423), respectively. Table 4 presents the output for the coefficient of determination for the various tested models. The basic information provided by the FTIR spectra analyses of drugs and their possible changes due to chemical interactions with the medium or chemical substances therein were compared for AL. The FTIR spectra analyses for AL and the mix (i.e., with the investigated drugs - lamivudine and metronidazole) are presented in Figure 3. The observed peaks for AL alone were compared with the characteristic spectra features of pure artemether and lumefantrine from the literature. This confirms the identity and the co-formulation status without interaction between actives or the excipients in the tablet as the spectra peaks for both artemether and lumefantrine were evident (Musibau et al., 2016).  In this study, the AL spectra in Figure 2a presented broad peaks at 3462.54 cm-1 due to the artemether component attributed to the aliphatic (O-H) bending.  It also featured peak at 2937.48 cm-1 due to the aliphatic (C-H) stretching vibrations. The values here were found to be consistent with values for pure artemether spectra from the literature and the eFTIR software library. The AL sample exhibited broad peaks at 3419.79 cm-1due to aliphatic (C-H) stretching vibration and 2951.09 due to aliphatic (C-H) bending vibrations due to the lumefantrine entity. Previous study by Musibau and co-workers also confirmed the co-formulated status of the actives in AL (Musibau et al., 2016).This formed the baseline for the investigation of the effect of the co-pelletized studied drugs. A similar study conducted by Balaji and Kumari evaluating the formulation of immediate release pellets containing artemether and lumefantrine revealed artemether  and lumefantrine spectral bands that were consistent with the literature values also indicating that artemether and lumefantrine in their product were co-formulated without any form of interactions (Balaji and Kumari, 2013). The inference therefore was that there was no breakdown in the structure of artemether in the co-formulation of the AL products. Similarly, the   literature values for pure lumefantrine and the software library were in agreement with the characteristic spectra presentation of lumefantrine in the AL sampled in this study. As observed in Figure 2, the spectra of artemether in the AL drug showed a broad peak at the wavenumber 3462.54 cm-1 and 2937.48 cm-1 due to (O-H and (C-H) stretching vibrations, respectively. The corresponding (O-H) stretching vibrations due to lamivudine and metronidazole were shifted downwards to 3387.0 and 3408.22 cm-1, respectively with respect to the plain AL spectral bands. The reference artemether band had 3379 and 2947 cm-1, respectively. Comparing the wavenumber difference for the effect of lamivudine and metronidazole with respect to the reference artemether band, they had values of 8 and 9 cm -1, respectively. The same arguments hold for the difference in wavenumber of peak expression for (C-H) stretching as approximately 18 and 9 cm-1, respectively. The endoperoxide bridge (C-O-O) demonstrated a broad IR stretching at 890 - 820 cm-1, adjudging by the reference artemether band. The co-pelletization with lamivudine and metronidazole did not affect the (C-O-O) stretching of artemether as they revealed IR spectra band within that range (i.e., 871.82 and 881.47 cm-1, respectively). Similarly, the spectra for pure lumefantrine revealed a broad peak at 3394.72, 2951.82 and 1462.04 cm-1for (O-H) and (C-H) stretchings and for (C-H) bending, respectively (Figure 2). Comparing the spectra bands for AL alone revealed a co-formulated product of artemether and lumefantrine along with excipients with no complexation with respect to the lumefantrine component. Comparing the spectra bands for lamivudine and metronidazole when co-pelletized, shift in spectral wavenumber downwards of values 7 and 14 cm-1 were observed for (O-H) stretching due to the investigated drugs, respectively. Considering (C-H) stretching, the differences in band wavenumber values were 22 and 12 cm-1, respectively. The software adjudged the spectra on Figure 2B and 2C corresponding to lamivudine and metronidazole, respectively, as not having a significant shift in the spectral features of artemether or lumefantrine as presented in Figure 2A. CONCLUSION In this study, the presence of metronidazole or lamivudine in the admixture with AL has not significantly affected the FTIR spectra presentation of AL. There was no complex formed with AL by either lamivudine or metronidazole. AL tablet can be co-administered with either lamivudine or metronidazole, based on the in vitro assessments. Furthermore, the release profile of AL drug components was affected by the presence of metronidazole but unaffected by lamivudine. The release of artemether from AL matrix in the presence of lamivudine and metronidazole were first order kinetics and Higuchi respectively. The release kinetics for lumefantrine for both drugs was Higuchi. ACKNOWLEDGMENT The authors are grateful to the Management of Central Research Laboratory of University of Lagos for the use of their equipments and Mr. PD Ojobor for his technical assistance. AUTHOR’S STATEMENT The authors have no conflict of interest to disclose. Englishhttp://ijcrr.com/abstract.php?article_id=2425http://ijcrr.com/article_html.php?did=2425 Adjei GO, Kurtzhal JA, Rodrigues OP, Alifrangis M, Hoegberg LC, Kitche ED, Badoe EV, Klamptey R, Coka BD. 2008. Amodiaquine – artesunate vs Artemether – lumefantrine Efficacy and Safety Trial with One Year Follow up. Mal J, 7:127. Ahsan MR, Sultan MZ, Amjad FM, Sultana S, Baki MM. 2012. Ofloxacin with Paracetamol and Zinc in Aqueous Medium. J Sci Res, 4:201 – 708. Bennett PN, Brown MJ 2008. Clinical Pharmacology (10thedn) Church Hill Livingstone Elsevier, Edinburgh London, New York Oxford, Philadelphia, St. Louis, Syndney, Toronto Pp. 74 – 114. Bercu TE. 2007. Amoebic Colitis: New Insights into Pathogenesis and Treatment. Curr Gastroenterol Rep, 9(5): 429-33. CDC. 2008. HIV Transmission through Transfusion- Missouri and Colorado, 2008. Morbidity and Mortality Weekly Report, 59 (41):1335-9. https://www.ncbi.nlm.nih.gov/pubmed/20966896. (Accessed 12 January, 2015) Cohen MS, Hellmann N, Levy JA, DeCock K, Lange J. 2008. The Spread, Treatment, and Prevention of HIV-1: Evolution of a Global Pandemic. The J Clin Invest, 18 (4): 1244–54. Dey S, Dutta S, Mazumder B. 2012. Formulation and Evaluation of Floating Matrix Tablet of Atenolol for Gastroretentive Drug Delivery. Int J Pharm Pharm Sci, 3: 433-437. Edwards IR, Aronson JK. 2000. Adverse Drug Reactions: Definitions, Diagnosis and Management. Lancet, 356:1255 – 1259. Galia E, Nicolaides E, Horter D, Lobenberg R, Reppas C, Dressman JB. 1998 Evaluation of Various Dissolution Media for Predicting In Vivo Performance of Class I and II Drugs. Pharm Res, 15(5): 698-705. Gonzales ML. 2009. Antiamoebic Drugs for Treating Amoebic Colitis. Cochrane Database Syst Rev, 15(2): CD0060085. Huang L, Lizak PS, Javewardene AL, Marzan F, Lee MN, Aweeka FT. 2010. A modified method for determination of lumefantrine in human plasma by HPLC-UV and combination of Protein Precipitation by solid Phase Extraction: application to a Pharmacokinetic Study. Anal Chem Insight, 29(5): 15-23. Haque R, Huston CD, Hughes M, Houpt E, Petri WA. 2003. Current Concepts in Amoebiasis. N Engl J Med, 348: 1565-73. Kumari MH, Balaji A. 2013. Formulation and Evaluation of Immediate Release Pellets Containing Artemether and Lumefantrine. Int J Pharm Ind Res, 03(04): 335-345. Kumari MH. 2013. Recent Novel Advancements in Pellet Formulation: A Review. Int J Pharm Sci Res, 4(10): 3803-382. Lakka NS, Goswami N. 2012. Solubility and Dissolution Profile Studies of Gliclazide in Pharmaceutical Formulation by RP-HPLC. Int J Pharm, 3 (6): 126-129. Morris CA, Lopez-Lazaro L, Jung D, Methareethorn J, Duparc C, Borghini-Fuhrer I, Pokorny R, Shin CS, Fleckenstein L. 2012. A Study of Drug-Drug Interaction of Pyroaridine/artesunate and ritonavir in healthy volunteer. Am J Trop Med Hyg, 86(3):489-495. Mustapha MA, Iwuagwu MA, Uhumwangho MI. 2016. Comparison of Performance Indices of Artemether-lumefantrine Fixed Dose Combination Tablets using Quality by Design Approach 1: Physico-technical Metrics Evaluation. J  Pharm Sci Tech, 5(2):105-108. Rosenthal PJ. 2014. Protozoal and Helminthic Infection. In: Current Medical Diagnosis and treatment. Papadakis MA, McPhee SJ, Rabow MW (Eds), McGraw Hill Education, 53rd Ed. 1442-1477. Schooley RT. 2010. Antiretroviral Treatment of Adult HIV Infection. JAMA, 304 (3):321-333. doi 10,1001/jama.2010.1004. Sunesen VH, Pedersen BL, Kristensen HG, Mullertz A. 2005. In vivo / in vitro correlation for a poorly soluble drug, danazol using the flow through dissolution method with biorelevant dissolution media. Eur J Pharm Sci, 24: 305-313. Thompson MA, Aberg JA, Cahn P, Montaner SG, Rizzmadini G, Telenti A, Gatel JM, Gunthard HR, Hammer SM, Hirsch MS, Jacobsen DM, Reiss P, Richmann DD, Volberding PA, Yeni P, Vertzoni M, Dressman J, Buffer J, Thempenstall J, Reppas C. 2005. Simulating of Fasting Gastric Conditions and its Importance for the In-vivo Dissolution of Lipophilic Compounds. Eur J Pharm Biopharm, 60 (3): 413 – 417).  Wienka LC, Heath TG. 2005. Predicting In vivo Drug Interaction from In Vitro Drug Discovery Data. Nature, 4: 825-833. World Health Organization. 2015. Guidelines for the treatment of malaria, 3rd ed., WHO, Geneva 2015. http://www.who.int/malaria/publications/atoz/9789241549127/en/ (Accessed on September 10, 2015).

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241102EnglishN2018January19HealthcarePenile Agenesis- An Extremely Rare Urogenital Anomaly English3132Mayukh ChakrabortyEnglish Sanghamitra ChakrabortyEnglishAim: Penile agenesis, defined as aphallia, is an extremely rare congenital anomaly. Though this condition is diagnosed clinically, it may be associated with various ano-rectal and systemic malformations. Until date, very few cases of aphallia have been reported globally. The most important dilemma in such cases lies in the treatment associated with gender reassignment. Case Report: We report a rare of case aphallia as a solitary presentation, in a neonate born in a rural hospital of West Bengal and the dilemma imposed on the parents regarding the management. Discussion and Conclusion: The confusion regarding the surgical intervention associated with the psyco-sexual influence is a major hindrance. EnglishAphallia, Congenital anomaly, Ano-rectalIntroduction: Penile agenesis is a very rare genitourinary malformation noticed in 1 in 30 million live births.[1]Aphallia occurs from the absence or failure in development of genital tubercle. Aphallia was first reported by French Surgeon, Saviard in 1701 as “Child who had no rod”, but the detailed description was reported by Imminger in 1853. Literature review suggests that till date 100 cases have been reported.[2] Moreover, Penile agenesis may be solitary or associated with wide array of malformations like genitourinary, gastrointestinal tract and developmental defects of caudal axis.[3] In the present instance a solitary malformation of penile agenesis in a genotypic male new-born and the dilemma associated with the treatment is described. Case Report:                     A full term new born was delivered in a gynaecology and obstetric department of a rural hospital of West Bengal by emergency caesarean section. The baby was born of non-consanguineous marriage and the antenatal history of the mother was uneventful and uncomplicated. The birth weight of the child was 3.5 kg .On general examination of the child, there was no penis. The scrotum was well developed with bilateral normally descended testis. Urethral opening was noticed in the anterior aspect of scrotum. (Figure 1) No other external malformations were noticed. Ultrasonography of whole abdomen revealed no renal abnormality and no female internal genitalia was observed. Karyotyping workup confirmed that the baby was 46XY. No fistula or aberration of urethral tract was noticed in Cystourethrogram. The baby was diagnosed as a case of aphallia and the treatment of reconstruction surgery were discussed with the parent. The parent were counselled regarding the constraints of treatment in this rural set-up and transferred to a higher set-up for reconstruction surgery. Discussion: Aphallia develops as a result of failure of development of genital tubercle into phallus with incomplete separation of urogenital sinus from hindgut.[1] The urethral opening may be situated at sites like in the perineum, near pubis, anterior aspect of scrotum or most commonly just anterior to the anus and in the rectum. Penile agenesis associated with other malformations is much more prevalent (54%) than the solitary variety. In this instance we report a solitary variety of aphalia with urethral opening situated on the scrotum.[3]Aphallia must be differentiated from micropenis, rudimentary penis, penile amputation, concealed penis and pseudohermaphroditism.[4] The confusion lies in the surgical management of such patients. Genital reassignment surgery was the main stay of treatment consisting of bilateral orchiectomy, vaginal reconstruction, urethral transposition and labial reconstruction with adequate postoperative outcome. [5]Moreover, life-long hormonal therapy initiated during puberty is an important part of treatment. Recently phallus reconstruction has also been successful in few such cases.[6]So, there is great dilemma between both the treatment modalities. The newer concept of gender imprinting in brain since in-utero and the psychosocial consequences of gender reassignment is an important perspective in such cases.[7] Thus the treatment should be individualised according to age of presentation, psychosocial adjustment of both patient and parents. The treatment is a multidisciplinary approach involving a paediatric suregeon, urologist, endocrinologist and psychologist. Present case report is unique in this that the patient presented at birth and there are no other genitourinary malformations. The parents were counselled regarding both the surgical interventions and referred to a tertiary care centre. Moreover, recent advances in radiological techniques T2-Weighted MRI(Magnetic resonance imaging) can not only detect such anomalies at an early pregnancy but also ascertain the prognosis. But, it is not advisable in Indian scenario where the bias for male gender is still very high. Conclusion: This case highlights that though penile agenesis is a rare urogenital finding yet the main dilemma lies in the intervention. Thus, treatment should be individualised depending on the age of presentation and psychosexual orientation of patient. Pre-surgical counselling remains an indispensible part of the treatment. Acknowledgement: Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors/editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Conflict of interest and Funding: There is no conflict of interest and no funding from external agency. Englishhttp://ijcrr.com/abstract.php?article_id=2426http://ijcrr.com/article_html.php?did=2426 Kessler W.O., McLaughlin A.P., 3rd Agenesis of penis. Embryology and management. Urology. 1973;1:226–229. Jack S Elder. In: Walsh PC, Retik AB, Vaughan ED Jr, Wein AJ, (editors).Campbell’s Urology. 8th ed. Saunders: Elsevier Science; 2002. pp. 2343-45. Skoog S.J., Belman A.B. Aphallia its classification and management. J. Urol. 1989;141:589–592. Roth J.K.J., r M., arshall R.H., Angel J.R., Daftary M., Lewis R.W. Congenital absence of penis. Urology. 1981;17:579–583. Hendren W.H. The genetic male with absent penis and urethrorectal communication: experience with 5 patients. J. Urol. 1997;157:1469–1474.  Chibber PJ, Shah HN, Jain P, Yadav P. Male Gender Assignment in Aphallia: A Case Report and Review of theLiterature. Int Urol Nephrol 2005; 37: 317-19. Diamond M, Sigmundson HK: Sex reassignment at birth. Long term review and clinical implications. Arch Pediatr Adolesc Med 1997;151:298

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241102EnglishN2018January19HealthcareTethered Cord Syndrome - A Rare Case Report English3337Rohit Kumar JainEnglish Rattilal MeenaEnglish Sourav ShristiEnglish Neera SamarEnglish Yogesh Kumar MishraEnglish Satish KumarEnglish Rakesh SoniEnglish Ashish KhandelwalEnglish Adheer Kumar YadavEnglish Ashok YadavEnglishTethered cord syndrome is a stretch-induced functional disorder associated with the fixation (tethering) effect of inelastic tissue on the caudal spinal cord, limiting its movement. This abnormal attachment is associated with progressive stretching and increased tension of the spinal cord as a child ages, potentially resulting in a variety of neurological and other symptoms. Due to the variation of the growth rate of the spinal cord and the spinal column, the progression of neurological signs and symptoms is highly variable. We report a very rare case of young patient of tethered cord syndrome presented with incontinence of urine and stool since birth and progressive weakness of both lower limbs. Other associated features and radiological findings are also discussed. EnglishTethered cord syndrome, Occult spinal dysraphism, neurological, Urological, Incontinence, Low conus medullaris, MyelomeningoceleINTRODUCTION- A tethered spinal cord is best defined as an abnormal attachment of the spinal cord to the tissues that surround it. The term has acquired a number of different meanings over time. This label has been applied to descriptions of radiographic findings and to varied constellations of clinical signs and symptoms. For example, in 1976 Hoffman and colleagues [1] used the phrase ‘‘tethered spinal cord’’ to define a radiographic diagnosis---a spinal cord ‘‘with a low conus medullaris and a thickened filum terminale measuring 2 mm or more in diameter,’’ excluding other conditions such as ‘‘lipomyelomeningoceles, meningoceles, myelomeningoceles, diastematomyelia, and intraspinal space-occupying dysraphic conditions such as dermoid tumors, intraspinal meningoceles, neurenteric cysts, and teratomatous cysts’’[1] . Many of which today are considered typically representative of tethering of the spinal cord.                                                                                          TCS is an entity described as an array of congenital anomalies, including cutaneous, urologic, neurologic, and orthopedic systems. It is thought to result from the abnormal fixation of the distal spinal cord secondary to a developmentally acquired or post-operative pathology. Unlike spina bifida aperta (i.e., myelomeningocele), which is readily diagnosed prenatally, spina bifida occulta often manifests more insidiously. As a result, these closed defects are usually discovered following symptom onset or incidentally during an unrelated work up of coincident comorbidities. Because the prognosis of this syndrome is highly dependent on symptom duration, it is important that all disciplines potentially involved (urology, orthopedics, dermatology, pediatricians, etc.) be aware of this condition as delay in diagnosis can have serious long-term effects. CASE SUMMARY - A 19 year old male patient presented to us with complaint of incontinence of urine and stool since birth and weakness of both lower limbs from last 6 months. Incontinence of urine and stool was first noticed by his parents when he didn’t achieve control over urine and stool at the age when a normal child should have achieved. Weakness of both lower limbs was noticed by patient himself from last 6 months, which was intermittent and worsened after activities involving bending of spine. There was no history of seizures, headache and intellectual deterioration. Vital signs were normal. His general physical examination revealed a small 2×2 cm sized dimple over sacral region which had a tuft of hairs overlying it, foot deformity in the form of pes cavus and varus, single testis  ( right) since birth . Central nervous system examination revealed diminished motor power (3/5) into both lower limbs, Deep tendon reflex: knee/Ankle reflexes were diminished, Babinski sign was negative. Sensory system examination was normal. Other systemic examination was normal. Magnetic Resonance Imaging-Brain and spine revealed mild dilatation of bilateral lateral ventricles with normal third and fourth ventricles, T2W sagittal sequence reveals non-visualization of coccyx vertebrae and linear hypo-intense tract extending from skin to coccyx region. The lower end of the cord is reaching upto L3-L4 level. Magnetic Resonance Imaging-Abdomen and pelvis revealed small contracted Left sided undescended testis with thickened trabeculated wall of urinary bladder, possibly neurogenic bladder. The left testis is seen at the deep left inguinal canal and appears small in size, it measures approx 10 mm × 14 mm. DISCUSSION HISTORY: As early as the mid-19th century, there were descriptions of spinal cord tethering and related symptomatology. Johnson, in 1857, discusses a fatty sacral tumor connected with spinal membranes in a child. In 1891, in England, Jones performed the first successful intervention for tethered cord. In 1910, Fuchs observed incontinence with spinal flexion in myelomeningocele patients that was attributed to increased tension on the distal spinal cord[2,3]. While several other contemporaries discussed observations consistent with tethered cord, it wasn’t until 1976 that the term “tethered spinal cord” finally emerged as a designation; Hoffman et al. coined the phrase to describe a series of 31 surgical patients with an abnormally low lying conus medullaris and thickened filum (>2 mm) whose symptoms improved following sectioning of the filum[1]. Several authors have attempted to determine what constitutes a “normal” conus level. Their findings vary from the T12 to the inferior aspect of L2, with the most common termination at or above L1/L2 [4-8]. For the remainder of this paper, normal conus will be considered one that terminates at or above the L2 vertebral body. EMBRYOLOGY The spinal cord forms as the result of two distinct processes: primary and secondary neurulation. Primary neurulation entails the proliferation and folding of neuroectoderm into a neural tube that ultimately comprises the spinal cord. This process begins on postovulatory day (POD) 18; the notochord induces the overlying ectoderm to proliferate as neuroectoderm, forming a groove that progressively elevates until it fuses and forms the neural tube. Cutaneous ectoderm (which eventually becomes skin) separates from the neuroectoderm and fuses on the midline during a critical process called “disjunction.” The mesoderm forms the posterior bony and soft tissue elements. Disruption of this stage is responsible for many spinal cord pathologies, including, myelomeningocele (nondisjunction), lipomyelomeningocele (premature disjunction), and dermal sinus tract (incomplete disjunction). Closure of the neural tube begins around POD 22 at the site of future cervical levels; it precedes both rostrally and caudally. Closure of the rostral and caudal neuropore occurs by POD 26 and POD 28, respectively. The formation of the brain and the spinal cord mark the end of primary neurulation. Developmental failure of the primary neurulation process may result in an open neural tube defect.                                                               Secondary neurulation refers to the formation of distal spinal elements caudal to S2 as well as the filum. This phase of development occurs between POD 28 and 48. Neuroectoderm caudal to the posterior neuropore, also known as the “caudal cell mass (CCM),” begins canalization. During this process, the vacuoles that form in the middle of the CCM coalesce with the vacuoles located in the neural tube’s central canal [9]. Subsequently, disproportionate growth rates between the spinal cord and the vertebral column cause the spinal cord to ascend and pull away from its sacral attachments. The cauda equine forms as nerve roots elongate to accommodate the differential growth. This process of retrogressive differentiation continues until the conus reach the adult level by three months of age. Errors that occur during canalization or regression are thought to contribute to the formation of low lying conus, terminal lipomas/myelocystoceles and fatty filum pathology [9-11]. PATHOPHYSIOLOGY Our understanding of TCS pathophysiology has primarily been built upon the work conducted by Yamada et al. over the last 30 years. The authors postulated that progressive low to moderate traction placed on the filum causes a reduction of cytochrome a, a3, which indicates an ischemic state. Using animal models, Yamada’s group showed that the degree of caudal traction on the spinal cord correlated with the severity of neurological deficit secondary to the impairment of oxidative metabolism [12]. They also demonstrated a proportional reduction in spinal cord blood flow in relation to the force of traction, which they designated as “traction induced hypoxia” [12]. Their model also showed how chronic tension can preload the cord in such a way that even minor additional traction can cause severe, permanent damage [13]. A study conducted by Stetler et al. showed that the tethering of the filum terminale caused a reduction in the blood flow to the spinal cord, leading to tissue hypoxia as a result of mitochondrial redox dysfunction [14]. Metabolic derangements were corrected following restoration of blood flow; however, if blood flow was interrupted for longer periods of time, recovery was only partial. Under these circumstances, it appears that excessive tension can cause irreversible changes and permanent dysfunction that will not be restored following tethered cord release. CLINICAL PRESENTATION Common presenting signs and symptoms include cutaneous signatures associated with occult spinal dysraphism (OSD) (59%); neurogenic bladder (18%), lower extremity weakness, numbness, or spasticity (12%), leg or foot discrepancy (6%), foot deformity, spinal deformity, and non-dermatomal leg/back pain (6%) [15]. While children often present with a combination of findings, symptoms can also be isolated to one system. This diverse presentation is one of the reasons why it is so crucial for physicians to acquaint themselves with the clinical picture of TCS. The diagnosis of various syndromes should also encourage physicians to evaluate patients for OSD, as the two are often associated with each other.                 Significant cutaneous lesions can be seen in up to 3% of the general population; in patients with OSD, the incidence approaches 80% and there is a greater chance that multiple lesions will be detected upon careful examination [11,16,17]. At times, these findings may be the only symptoms indicating underlying dysraphism. Cutaneous discrepancies include midline hairy patches, hemangiomas, dermal pits/sinuses, hypertrichosis, subcutaneous lipoma, “cigarette burns,” lumbosacral appendage, and nevi. While the appearance of any of the aforementioned findings is sufficient to warrant investigation, recent work spanning 12 years of pediatric patients suggests that an isolated sacral dimple in an otherwise asymptomatic child has a significantly low association with tethered cord syndrome; the incidence of necessary surgical de-tethering in that population ranged from only 0.13 to 0.17% [18-20]. That being said, ultrasound is simpler to perform and easier to obtain than an Magnetic Resonance Imaging at a later point when the child may already have become symptomatic and will also require sedation.                 Urologic dysfunction is most commonly the initial derangement in OSD-related tethering. Patients encounter problems that range from blatant incontinence to subtle changes observed during urodynamic studies. Presentation may include incontinence, urinary urgency, increased urinary frequency, and recurrent UTIs; in the pediatric population, these symptoms tend to be more subtle than other clinical findings [14]. Because bladder dysfunction is difficult to assess in infants, these problems may not even become apparent until children are much older. The most common bladder symptom among toddlers is delayed or unsuccessful toilet training; during testing, detrusor hyperreflexia is the most common finding. Because a disruption in urodynamics often precedes clinical symptoms, this highlights the importance of urological work to aid in preventing delayed diagnosis and treatment. Besides detrusor hyperreflexia, other common symptoms include diminished bladder compliance, external detrusor sphincter dyssynergia, decreased sensation, and hypocontractile destrusor function [21,22].                  Neurological problems that manifest in TCS involve the disruption of the motor and sensory pathways of the lower extremities. Although they comprise elements of upper and lower motor neuron dysfunction, motor deficits are more prevalent than sensory deficits [14]. Sensory deficits, if present, are in the feet or perineum or children may present with painless ulcerations of the feet/legs.                 Orthopedic abnormalities are found in more than 90% of patients with TCS [14]. Among children, foot deformities are most common. These deformities most likely result from neuromuscular imbalance at a time when bones are growing and aligning; deformities are unlikely to arise later in life if mal-alignment does not occur during this period. Other abnormalities include limb length discrepancies, gluteal asymmetry, vertebral anomalies, and scoliosis. Progressive scoliosis or kyphosis can be seen in about 25% of children with TCS and may also contribute to complaints regarding pain. Vertebral anomalies are commonly observed in children with TCS. These include bifid vertebrae, laminar anomalies, hemivertebrae and sacral agenesis. Segmentation errors may be multiple and these boney abnormalities can be observed in approximately 95% of children with TCS [11]. DIAGNOSIS                  Diagnosis of TCS requires the correlation of clinical symptoms with relevant radiographic findings. Presently, to our knowledge, there has never been a case of TCS that was reported with normal imaging. Among the various radiographic procedures available, plain X-rays have the most limited application; they are primarily used to follow the progression of scoliosis. Ultrasound is ideal for infants because there is no need for radiation or sedation. It is also reported to have 96% sensitivity and 96% specificity [18]. That being said, ultrasound is limited by operator abilities and often difficult to interpret. Additionally, its use is restricted to infants 4-6 months old because spine ossification reduces the reliability of ultrasound findings [18,23]. It can, however, function as a screening tool. If ultrasound results are normal in the setting of sacral dimples or isolated strawberry hemangioma, then the probability of TCS is relatively low and Magnetic Resonance Imaging studies can be postponed [24]. Magnetic Resonance Imaging is the imaging procedure of choice for the assessment of OSD/TCS. T1-weighted imaging provides clear anatomical detail of neural tissue and the filum. This enables visualization of vertebral levels, the conus position, and the presence of fat/thickening/syrinx. Sagittal views allow for level localization while axial views display fat and the diameter of the filum. As mentioned before, a filum below the L2 vertebral body or thicker than 2 mm is considered abnormal in children. The absence of movement between supine and prone MR imaging would also indicate a diagnosis of TCS [1]. MR imaging can also reveal urologic sequelae, such as a distended bladder. T2-weighted imaging permits the identification of spinal cord tumors and fluid containing structures. Complete imaging of the entire neuroaxis is important for the screening of “skip lesions” or other abnormalities that are often observed in association with TCS (i.e., Myelomeningocele, Split cord malformation, dermal and lipomatous tumors, etc.)               Using only medical history and examination to determine the cause of bladder dysfunction in children can be problematic without the assistance of urodynamic studies (UDS). Because TCS symptom reversal is associated with the duration of dysfunction, patients will have a better chance for successful outcomes if urodynamic studies are implemented early to help establish a definitive diagnosis. UDS can also indicate clinical deterioration and provide a way for physicians to monitor patient improvement following a detethering procedure. For these reasons, physicians should obtain UDS prior to and following any surgical procedure. The most common UDS finding is detrusor hyperreflexia. The following metrics may also be identified: decreased bladder compliance, dyssynergia, and decreased sensation. Important aspects of bladder function assessment include: bladder capacity, bladder pressure, leak point pressure, compliance, uninhibited contractions, EMG activity, and sensation [10]. CONCLUSION The prompt identification of TCS still represents a major clinical challenge. Some of the obstacles contributing to that challenge are symptom ambiguity, presentation discrepancy, and the absence of a standardized pathology accounting for the majority of OSD cases resulting in pediatric TCS. These factors are particularly problematic with respect to timely diagnosis and treatment of children. Because the duration of tethering is a key determinant in disability and recovery, it is paramount that physicians from various specialties establish a uniform understanding of the physiology, presentation, classification, and treatment of OSD. Additionally, given that presentation of OSD can be limited to subtle changes in urodynamic studies, it is imperative that a dedicated multidisciplinary team of specialists (including pediatric urologists, pediatric neurosurgeons, pediatric orthopedic surgeons, and physiatrists) evaluate patients with suspected TCS. While the true incidence of OSD is unknown, advanced imaging, a greater clinical awareness, and ongoing relevant publications assist in making this condition a more mainstream diagnosis among primary care physicians. ACKNOWLEDGEMENT: Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. SOURCE OF FUNDING:      Nil CONFLICT OF INTEREST :    None Englishhttp://ijcrr.com/abstract.php?article_id=2427http://ijcrr.com/article_html.php?did=2427[1] Hoffman HJ, Hendrick EB, Humphreys RP. The tethered spinal cord: its protean manifestations, diagnosis and surgical correction. Childs Brain 1976;2(3):145–55. [2] Hertzler DA, DePowell JJ, Stevenson CB, Mangano FT (2010) Tethered cord syndrome: a review of the literature from embryology to adult presentation. Neurosurg Focus 29: e1. [3] Fuchs A (1910) Euber beziehungen der enuresis nocturna zu rdimentarformen der spina bifida occulta (myelodisplasie). Wien Med Wochenschr 80: 1569-1573 [4] McLone D, Warf B (2001) Pathophysiology of tethered cord syndrome Pediatric Neurosurgery: Surgery of the Developing Nervous system. (4th edn), 20: 282-288. [5] Drake J (2007) Surgical management of tethered spinal cord-walking the fine line. Neurosurg Focus 23: E8. [6] Pang D, Wilberger JE (1982) Tethered cord syndrome in adults. J Neurosurg 57: 32-47. [7] Wilson DA, Prince JR (1989) John Caffey award. MR imaging determination of the location of the normal conus medullaris throughout childhood. AJR Am J Roentgenol 152: 1029-1032. [8] Saifuddin A, Burnett SJ, White J (1998) The variation of position of the conus medullaris in an adult population. A magnetic resonance imaging study. Spine 23: 1452-1456. [9] Blout JP, Elton S (2001) Spinal Lipomas. Neurosurg Focus 10: e3. [10] Lew S, Kothbauer K (2007) Tethered cord syndrome: an updated review. Pediatr Neurosurg 43: 236 248. [11] Warder DE (2001) Tethered cord syndrome and occult spinal dysraphism. Neurosurg Focus 10: e1. [12] Yamada S, Zinke DE, Sanders D (1981) Pathophysiology of ‘tethered cord syndrome’. J Neurosurg 54: 494-503. [13] Yamada S (1996) Pathophysiology of the tethered spinal cord; in Yamada S (eds): Tethered cord syndrome. Park Ridge, American Association of Neurological Surgeons, 29-48. [14] Stetler WR, Park P, Sullivan S (2010) Pathophysiology of adult tethered cord syndrome: review of the literature. Neurosurg Focus 29: e2. [15] Bui CJ, Tubbs RS, Oakes WJ (2007) Tethered cord syndrome in children: a review. Neurosurg Focus 23: e2. [16] Powell KR, Cherry JD, Hougen TJ, Blinderman EE, Dunn MC (1975) A prospective search for congenital dermal abnormalities axis. J Pediatr 87: 744-750. [17] Chapman P, Stieg P, Magge S, Barnes P, Mei F (1999) Spinal lipma controversy. Neurosurgery 44: 186-192. [18]  Kucera J, Coley S, O’Hara S, Kosnik E, Coley B (2014) The simple sacral dimple: diagnostic yield of ultrasound in neonates. Pediatric Radiology 45: 211-216. [19]  Henriques JG, Pianetti G, Henriques KS, Costa P, Gusmão S (2005) Minor skin lesions as markers of occult spinal dysraphisms – prospective study. Surg Neurol 63: S8-S12. [20] Chern JJ, Kirkman JL, Shannon CN, Tubbs RS, Stone JD, et al. (2012) Use of lumbar ultrasonography to detect occult spinal dysraphism. J Neurosurg Pediatr 9: 274-279. [21] Fone PD, Vapnek JM, Litwiller SE, Couillard DR, McDonald CM, et al. (1997) Urodynamic findings in the tethered spinal cord syndrome: does surgical release improve bladder function? J Urol 157: 604-609. [22] Giddens JL, Radomski SB, Hirshberg ED, Hassouna M, Fehlings M (1999) Urodynamic findings in adults with tethered cord syndrome. J Urol 161: 1249-1254. [23] Rajpal S, Iskander B (2011) Lipomyelomeningocele. In: W Richard, Youmans Neurological Surgery. (6thedn), Philadelphia, PA. Elsevier Inc. [24] Kesler H, Dias MS, Kalapos P (2007) Termination of the normal conus medullaris in children: a whole-spine magnetic resonance imaging study. Neurosurg Focus 23: e7. [25]Derek C. Samples and Izabela Tarasiewicz (2016)Review and Classification of Occult Spinal Dysraphism and Tethered Cord Syndrome in Children  ; J Spine, an open access journal Volume 5 • Issue 4 • 1000325