Genomic Analysis of an Indian G8P[1] Caprine Rotavirus-A Strain Revealing Artiodactyl and DS-1-Like Human Multispecies Reassortment.

The surveillance studies for the presence of caprine rotavirus A (RVA) are limited in India, and the data for the whole-genome analysis of the caprine RVA is not available. This study describes the whole-genome-based analysis of a caprine rotavirus A strain, RVA/Goat-wt/IND/K-98/2015, from a goat kid in India. The genomic analysis revealed that the caprine RVA strain K-98, possess artiodactyl-like and DS-1 human-like genome constellation G8P[1]-I2-R2-C2-M2-A3-N2-T6-E2-H3. The three structural genes (VP2, VP4, and VP7) were close to caprine host having nucleotide-based identity range between 97.5 and 98.9%. Apart from them, other gene segments showed similarity with either bovine or human like genes, ultimately pointing toward a common evolutionary origin having an artiodactyl-type backbone of strain K-98. Phylogenetically, the various genes of the current study isolate also clustered inside clades comprising Human-Bovine-Caprine isolates from worldwide. The current findings add to the knowledge on caprine rotaviruses and might play a substantial role in designing future vaccines or different alternative strategies combating such infections having public health significance. To the best of our knowledge, this is the first report on the whole-genome characterization of a caprine RVA G8P[1] strain from India. Concerning the complex nature of the K-98 genome, whole-genome analyses of more numbers of RVA strains from different parts of the country are needed to comprehend the genomic nature and genetic diversity among caprine RVA.

Diffusion-weighted MRI assessment of the peritoneal cancer index before cytoreductive surgery.

BACKGROUND: Patients with limited peritoneal metastases from colorectal cancer may be candidates for an aggressive surgical approach including cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (CRS-HIPEC). Selection is based on surgical inspection during laparoscopy or laparotomy. The aim of this study was to investigate whether diffusion-weighted MRI (DW-MRI) can be used to select patients for CRS-HIPEC. METHODS: This was a prospective study at a tertiary referral centre. Patients with confirmed or suspected colorectal peritoneal metastases scheduled for exploratory laparotomy or laparoscopy were eligible. Two radiologists assessed the peritoneal cancer index (PCI) on CT (CT-PCI) and DW-MRI (MRI-PCI). The reference standard was PCI at surgery. Radiologists were blinded to the surgical PCI and to each other's findings. The main outcome was the accuracy of DW-MRI in predicting whether patients had resectable disease (PCI less than 21) or not. RESULTS: Fifty-six patients were included in the study, of whom 49 could be evaluated. The mean(s.d.) PCI at surgery was 11·27(7·53). The mean MRI-PCI was 10·18(7·07) for reader 1 and 8·59(7·08) for reader 2. Readers 1 and 2 correctly staged 47 of 49 and 44 of 49 patients respectively (accuracy 96 and 90 per cent). Both readers detected all patients with resectable disease with a PCI below 21 at surgery (sensitivity 100 per cent). No patient was overstaged. The intraclass correlation (ICC) between readers was excellent (ICC 0·91, 95 per cent c.i. 0·77 to 0·96). MRI-PCI had a stronger correlation with surgical PCI (ICC 0·83-0·88) than did CT-PCI (ICC 0·39-0·44). CONCLUSION: DW-MRI is a promising non-invasive tool to guide treatment selection in patients with peritoneal metastases from colorectal cancer.

Anterior Dislocation of Elbow with Neurovascular Injury: A Rare Case Report.

INTRODUCTION: Anterior elbow dislocation is an infrequent lesion, produced by direct trauma to the proximal ulna after a fall on the elbow in flexion and injury to the neurovascular bundle is not infrequent. Authors report a case of acute anterior dislocation of the elbow joint with neurovascular injury. CASE REPORT: A 30-year-old male admitted with a history of accidental fall followed by pain, swelling over his left elbow. Clinical, radiological, Doppler investigations revealed anterior dislocation of the elbow with brachial artery injury with posterior interosseous nerve palsy without any bony injury. Immediate closed reduction, primary vascular repair with fasciotomy was done following which elbow function improved. CONCLUSION: Anterior dislocations of elbow joint are among the rarest of injuries. Because the dislocation is anterior, injury to nerve and vessel can occur frequently. Therefore, a careful assessment for neurovascular injury mandatory. Early proper reduction and management of neurovascular injury if any is necessary for good elbow function.

A pharmacological overview of lamotrigine for the treatment of epilepsy.

INTRODUCTION: Epilepsy is one of the most common neurological disorders, affecting about 2% of the population worldwide. Lamotrigine (LTG) is a second generation anti-epileptic drug (AED) with broad spectrum of activity, a favourable side-effect profile, simpler dosing than earlier drugs and efficacious in diverse epilepsy syndromes. Areas covered: The present review focuses on pharmacodynamics, pharmacokinetics, clinical efficacy, safety and tolerability of LTG and its effect on cognition, psychiatry, quality of life, women and pregnancy along with effect of enzyme inducing and enzyme inhibiting drugs over LTG and their effect on serum level fluctuations by collecting data from various studies over the years until 2016. Expert commentary: Results from various studies and clinical trials indicate that LTG possessed a favourable profile of anticonvulsant activity and good tolerability as a monotherapy/or add-on therapy in children and adult patients against several types of seizures and syndromes. It has wide clinical dose range with favourable pharmacokinetic properties making it an excellent therapeutic option in epilepsy.

Performance of submerged membrane bioreactor (SMBR) with and without the addition of the different particle sizes of GAC as suspended medium.

In this study the effect of different particle sizes of granular activated carbon (GAC) on the performance of a submerged membrane bioreactor (SMBR) was investigated. The sizes of GAC used were 150-300, 300-600 and 600-1200 µm. The SMBR was operated at a filtration flux of 20 L/m(2)h. The removal of dissolved organic carbon (DOC) and chemical oxygen demand (COD) with the addition of GAC was 95%. The concentration of biopolymers, humic, building block and low molecular weight neutral and acids in the SMBR effluent was reduced by 20%, 66-76%, 20-50%, 30-56%, respectively. It helped to reduce the sludge volume index (SVI) and transmembrane pressure (TMP) development by 30-40% and 58%, respectively. However, the removal of NH4(+) and PO4(3-) was relatively low of 35-45% and 34-43%, respectively. The SMBR effluent was rich in PO4(3-) and was removed/recovered using hydrated ferric oxide (HFO). The removal of PO4(3-) was almost 90%.

Formulation and Evaluation of Hydroxypropyl Methylcellulose-based Controlled Release Matrix Tablets for Theophylline.

The objectives of the study were to formulate hydroxypropyl methyl cellulose-based controlled release matrix tablets for theophylline with varying drug:polymer ratios (1:1 and 1:2) and differing tablet hardness (5, 6 and 7 kg/cm(2)), and to evaluate the tablet's physico-chemical properties such as hardness, uniformity of weight, friability, drug content and in vitro drug release. Initially, granules were made by wet granulation technique and evaluated for angle of repose, bulk density, tapped density, bulkiness, compressibility index and hausner ratio. The results indicate good flow property of the granules and thus, the evaluated tablet physical properties were within the acceptable limits. The FT-IR study for the F-6 formulation showed that there was no interaction between the drug and the polymer. In vitro release studies were performed using Disso-2000 (paddle method) in 900 ml of pH 7.4 at 50 rpm. The result indicated that at high drug:polymer ratio (1:2) and hardness value 7 kg/cm(2), prolonged drug release was observed than the low drug: polymer ratio (1:1) and hardness values (5 and 6 kg/cm(2)). The release kinetics was found to follow korsmeyers-peppas model and the mechanism of drug release was by non-fickian or anomalous diffusion. The F-6 formulation was chosen for stability studies. F-6 formulation was stable when it was kept at different temperatures for a period of 6 months.

Controlled release of 2, 3 desulfated heparin exerts its anti-inflammatory activity by effectively inhibiting E-selectin.

Control of inflammation using appropriate anti-inflammatory agent prevents wound from becoming chronic. Heparin is a heterogeneous mixture of polysaccharide molecules with a mean molecular weight between 12-30 kDa containing 200-300 disaccharide units of glycosaminoglycan chains. Chemical modifications leading to generation of a unique pentasaccharide sequence effectively reduces its anticoagualant activity, while retaining its anti-inflammmatory property. In this study, Standard heparin was partially desulfated to 2, 3 desulfated heparin (2, 3 DSH) and its anti-inflammatory property was determined by assessing its ability to prevent static adhesion of leukocytes to endothelium and clotting assay. The effectiveness of 2, 3 DSH to down regulate E-selectin and key proinflammatory cytokines (IL-1beta and IL-6) was assessed by western blot and immunocytochemistry. These results were compared with commercially available 2-Desulfated Heparin (2DSH) and standard heparin (SH). Finally, a controlled delivery system of 2, 3 DSH was designed using chitosan microspheres and collagen as scaffold. Optimal loading of 2, 3 DSH was achieved and the release kinetics were tuned to follow a controlled release pattern. The steady state concentration of 2, 3 DSH was found to be optimal to elicit anti-inflammatory property and could achieve inhibition of E-selectin expression while unaffecting the normal clotting cascade.

Formulation and Evaluation of Theophylline Controlled Release Matrix Tablets using Guar gum

Theophylline controlled release matrix tablets were prepared with guar gum in two ratios and with three different hardness of 5, 6 and 7kg/cm2. Theophylline controlled release granules were prepared and evaluated for the angle of repose, bulk density, tapped density, compressibility index and hausners ratio. All the formulation showed good flow properties. The compressed tablets were evaluated for the hardness, uniformity of weight, friability, drug content and invitro dissolution studies. All the formulations showed compliance with pharmacopial standards. There was no interaction between drug, polymer and other excipients. It was confirmed by FTIR studies. Among all the formulations F6 (i.e. polymer ratio1:2 and hardness 7kg/cm2) showed prolong release when compare to other formulations. The drug release kinetics showed zero order. The optimum formulation (F6) was stable when it was stored at 40 + 20 C, 280 + 20 C and at 450 + 20 C for 6 months(AU)

Functionally modified gelatin microspheres impregnated collagen scaffold as novel wound dressing to attenuate the proteases and bacterial growth.

An attempt was made to develop a new therapeutic delivery system which would play a dual role of attenuating MMP activity in the wounds and also prevent infection by controlled delivery of antimicrobials. A catechol type MMP inhibitor 2,3-dihydroxybenzoic acid (DHBA) was conjugated to gelatin microspheres using EDC/NHS as coupling agents. The potential of the modified gelatin microspheres (DHB-MS) to attenuate the proteases such as MMP 2 and MMP 9 in the diabetic wound tissues was investigated by gelatin zymography. Further the modified microspheres were loaded with doxycycline and impregnated in a reconstituted collagen scaffold as novel wound dressing. The in vitro release behavior of doxycycline from both DHB-MS and DHB-MS impregnated collagen scaffold was investigated. DHB-MS when incubated with the tissue lysate for 6h displayed the complete inhibition of the MMPs in the tissue lysate. The in vitro drug release studies from the collagen scaffold exhibited the burst release of 44%, exerted immediate chemo prophylaxis and sustained delivery for 72 h. The MTT assay and fluorescent labeling with calcein AM indicated that the DHB-MS is biocompatible to human foreskin fibroblasts. Thus the system developed provides wider scope to control the pathogens involved in infection and also the excess matrix degradation.

Diseño de una matriz de soporte compuesta de colágeno de piel de tiburón-aloe para ingeniería tisular / Design of shark skin collagen-aloe composite scaffold for tissue engineering

Se ha demostrado que el colágeno es un nuevo biomaterial utilizado para la administración de fármacos, la fabricación de apósitos o como sustrato para ingeniería tisular cuya biocompatibilidad y propiedades biodegradables son únicas. El colágeno bovino y porcino tipo I constituyen una fuente fácilmente disponible de material de soporte para diversas aplicaciones biomédicas. Sin embargo, estas fuentes conllevan cierto riesgo potencial de enfermedades infecciosas como la encefalopatía espongiforme bovina o la encefalopatía espongiforme transmisible. Por esta razón, existe una demanda de colágeno tipo I procedente de otras fuentes. En el presente estudio, se utilizan animales acuáticos y, en concreto, especies de tiburón en las que el colágeno tipo I es una proteína principal de la piel y la estructura tiene similitud con la de las especies mamíferas. Se ha intentado utilizar colágeno de piel de tiburón como matriz de soporte con extracto de aloe para mejorar la estabilidad. Estas matrices de soporte se caracterizaron por varias propiedades fisicoquímicas y por la evaluación de biocompatibilidad para facilitar el crecimiento de fibroblastos dérmicos humanos in vitro. La incorporación de extracto de aloe influyó enormemente en la morfología y las propiedades fisicoquímicas de la matriz de soporte. Se observó in vitro que los fibroblastos conservaban la orientación organizada en forma de huso al cultivarse sobre la matriz de soporte de colágeno. Así, la matriz de soporte de colágeno desarrollada con una proporción de 10:1 de colágeno de piel de tiburón y extracto de aloe, respectivamente, sirvió como material biocompatible con una resistencia a la tracción apreciable. La investigación anterior sugiere que la matriz de soporte de colágeno de piel de tiburón desarrollada puede ser una alternativa efectiva al colágeno de mamífero en el campo de la ingeniería tisular y para diversas aplicaciones en la curación de heridas (AU)

Collagen has proven to be a novel biomaterial used for drug delivery, wound cover dressings or as a substrate for tissue engineering with unique biocompatibility and biodegradable properties. Bovine and porcine Type I collagen provide a readily available source of scaffold material for various biomedical applications. However these sources have some potential risk of infectious diseases such as bovine spongiform encephalopathy or transmissible spongiform encephalopathy. Hence there is demand for an alternative Type I collagen from various other sources. The present study utilizes the aquatic animals particularly the shark species in which collagen Type I is a major protein in the skin and the structure has similarity to that of mammalian species. An attempt was made to use shark skin collagen as scaffold with the extract of aloe to improve the stability. These scaffolds were characterized for various physicochemical properties and biocompatibility assessment to support the growth of human dermal fibroblasts in vitro. The incorporation of aloe extract highly influenced the morphology and physicochemical properties of the scaffold. It was observed in vitro that the fibroblasts retained the spindle shape, organized orientation when cultured over collagen scaffold. Thus the developed collagen scaffold at 10: 1 ratio of shark skin collagen and aloe extract respectively served as a biocompatible material with appreciable tensile strength. The above investigation suggests that the developed shark skin collagen scaffold could be an effective alternative for the mammalian collagen for tissue engineering and various wound healing applications (AU)

Formulación y evaluación de comprimidos matriciales de clorhidrato de ambroxol, usando un polímero natural hidrofílico / Formulation and Evaluation of Matrix tablets of Ambroxol hydrochloride using Natural hydrophilic polymer

Se prepararon comprimidos matriciales de liberación prolongada de clorhidrato de ambroxol con diversas proporciones de fármaco: polímero tales como F-1(1:1), F-2(1:1,5) y F-3 (1:2). Se utilizó goma xántica para la formación de la matriz y celulosa microcristalina como diluyente. Se prepararon y evaluaron gránulos para determinar la densidad aparente sin compactar, la densidad compactada, el índice de compresibilidad, el índice de Hausner y el ángulo de reposo. Todos los gránulos se lubricaron y comprimieron con punzones planos de 9 mm. Los comprimidos se evaluaron para determinar la uniformidad de peso, el contenido de principios activos, la friabilidad, la dureza y la disolución in vitro. Todas las formulaciones se ajustaron a los estándares farmacopeicos. F-3 mostró una liberación prolongada de fármaco durante 12 horas con una liberación del 97,3% y el perfi l de liberación fue similar al de la muestra de clorhidrato de ambroxol comercial (A-MS). Además, se realizaron estudios de estabilidad según la guía ICH. La liberación de fármaco sigue cinéticas de orden cero (0,9661) y se determinó que el mecanismo era difusión combinada con erosión (AU)

Sustained release matrix tablets of ambroxol hydrochloride of different drug: polymer ratios, such as F-1(1:1), F- 2(1:1.5) and F-3 (1:2). Xanthan gum was used as matrix former and microcrystalline cellulose was used as diluent. Granules were prepared and evaluated for loose bulk density, tapped density, compressibility index, hausners ratio and angle of repose. All the granules were lubricated and compressed using 9mm fl at-faced punches. Compressed tablets were evaluated for uniformity of weight, content of active ingredient, friability, hardness and In-vitro dissolution. All the formulations showed compliance with Pharmacopoeial standards. F-3 showed the sustained release of drug for 12 hours with 97.3% release and the release profi le was close to the marketed sample of ambroxol hydrochloride (A-MS) and Stability studies were performed as per ICH guide. The drug release follows zero order kinetics (0.9661) and the mechanism was found to be diffusion coupled with erosion (AU)

Cellular and cytokine responses in feathers of chickens vaccinated against Marek's disease.

In Marek's disease virus infection, feather follicle epithelium (FFE) constitutes the site of formation of infectious virus particles and virus shedding. The objective of this study was to characterize cellular and cytokine responses as indicators of cell-mediated immune response in FFE and associated feather pulp following immunization against Marek's disease. Analysis of feather tips collected between 4 and 28 days post-immunization (d.p.i.) from chickens vaccinated post-hatch with either CVI988/Rispens or herpesvirus of turkeys revealed that replication of these vaccine viruses started at 7d.p.i., peaked by 21d.p.i., and subsequently, showed a declining trend. This pattern of viral replication, which led to viral genome accumulation in feather tips, was associated with infiltration of T cell subsets particularly CD8+ T cells into the feather pulp area and the expression of cytokine genes such as interferon-gamma, which is an indication of elicitation of cell-mediated immune responses at the site of virus shedding.

A fluorescent Gram stain for flow cytometry and epifluorescence microscopy.

The fluorescent nucleic acid binding dyes hexidium iodide (HI) and SYTO 13 were used in combination as a Gram stain for unfixed organisms in suspension. HI penetrated gram-positive but not gram-negative organisms, whereas SYTO 13 penetrated both. When the dyes were used together, gram-negative organisms were rendered green fluorescent by SYTO 13; conversely, gram-positive organisms were rendered red-orange fluorescent by HI, which simultaneously quenched SYTO 13 green fluorescence. The technique correctly predicted the Gram status of 45 strains of clinically relevant organisms, including several known to be gram variable. In addition, representative strains of gram-positive anaerobic organisms, normally decolorized during the traditional Gram stain procedure, were classified correctly by this method.

Mapping of the cellular immune responses to woodchuck hepatitis core antigen epitopes in chronically infected woodchucks.

T-cell responses to hepatitis B virus nucleocapsid antigens (HBcAg and HBeAg) play an important role in disease outcome in those infected with hepatitis B virus (HBV). The woodchuck is naturally infected in the wild with woodchuck hepatitis virus (WHV), which shows a high degree of genetic homology to HBV and produces a similar pattern of infection in its natural host. Twenty-three overlapping peptides were constructed to cover the entire WHV core region and used to identify immunodominant cellular epitopes in the nucleocapsid antigen using peripheral blood lymphocytes from 12 chronic WHV carrier and 4 uninfected control animals. A peripheral blood lymphocyte response was seen in all of the chronic WHV carrier animals to at least one peptide, and in 8 of the 12 chronic carrier animals a response was observed to 5 common peptides: peptide analogues of amino acids 16-30, 38-52, 50-69, 76-90 and 91-105. Peptide 91-105 produced maximal proliferation in 5 out of 12 infected animals. In addition, a difference in response was observed between wild and laboratory infected animals; the latter appeared to have a lower response to peptides than animals infected in the wild. This study provides evidence that the woodchuck has a population of peripheral blood cells which are sensitised to epitopes within the nucleocapsid protein and provides a basis on which to develop the use of the woodchuck as an immunological model of HBV infection for testing therapeutic means of enhancing this response.

Serum proteins in hepatic amoebiasis.

Serum proteins were estimated in two patient groups, namely (i) 20 cases with proven ameobic liver abscess and (ii) 12 cases with hepatic amoebiasis without demonstrable pus, commonly referred to by the misnomer 'ameobic hepatitis'. In amoebic liver abscess a fall in albumin and an increase in alpha 2 and gamma globulin results in a serum protein pattern somewhat different from that in hepatic amoebiasis without demonstrable pus, where an elevation in gamma globulin was infrequent. Post-therapy serum protein estimations revealed that, irrespective of the presence or absence of pus, a rising gamma globulin level in hepatic amoebiasis may be of prognostic significance and post-treatment surveillance would be necessary in patients showing this type of response.