An Uncommon Ischial Tuberosity Tuberculosis Infection.

Introduction: Ischium is one of the rare sites to be involved by mycobacterium tuberculosis. The incidence is generally not more than 0.2% in any of the large series. We report an unusual case of extrapulmonary tuberculosis of the ischial tuberosity presenting with chronic gluteal pain of 6 months duration. Case Report: A 35-year-old male patient presented with chronic dull aching gluteal pain of 6 months duration, for which lifestyle modifications and rest were advised initially. Antituberculosis chemotherapy was administered (for a period of 1 year) following histopathological confirmation of tuberculosis. At 1 year post antitubercular therapy, the patient had no pain and was symptom free. Furthermore, radiographs showed healed right ischial tuberosity osteomyelitis. Conclusion: Tuberculosis involving the ischial tuberosity is rare. The early diagnosis is mandatory for good results, and with a worldwide resurgence of the disease, a high index of suspicion is necessary. Prompt diagnosis and treatment resulted in a good clinical outcome in this patient.

Biological Synthesis of PbS, As3S4, HgS, CdS Nanoparticles using Pseudomonas aeruginosa and their Structural, Morphological, Photoluminescence as well as Whole Cell Protein Profiling Studies.

Metal sulfide nanoparticles are semi-conductors that possess many applications in optics, optoelectronics and magnetic devices. There are physical and chemical methods for their synthesis but such methods involve toxic precursors as well as many obnoxious by-products. Hence, biological synthesis of metal sulfide nanoparticles are efficient enough to transform toxic metals to non-toxic ones. Pseudomonas aeruginosa, isolated from textile effluent and tolerant of high levels of heavy metals, was used for the green synthesis of metal sulfide (HgS, As3S4, CdS and PbS) nanoparticles. The optical, structural and morphological nature of metal sulfide nanoparticles was also determined. FTIR (Fourier Transform Infra-red) analysis showed spectral changes when P. aeruginosa was grown in medium containing heavy metals viz. Hg, As, Pb and Cd indicating that there are functional groups viz. carboxyl, hydroxyl, phosphate, amino and amide, that exists on the surface of the bacteria, thus facilitating binding of metals on its surface. The bacterial samples which were treated with different metals at different concentrations, were subjected to whole cell protein analysis using SDS-PAGE (Sodium dodecyl Sulphate- Polyacrylamide gel electrophoresis) and protein profiling. The total protein estimation revealed that there was an increase in the protein concentration in the presence of heavy metals and a significant change in the banding pattern was observed which showed induction of a set of proteins under heavy metal stress especially mercury.

Pentavalent Sialic Acid Conjugates Block Coxsackievirus A24 Variant and Human Adenovirus Type 37-Viruses That Cause Highly Contagious Eye Infections.

Coxsackievirus A24 variant (CVA24v) and human adenovirus 37 (HAdV-37) are leading causative agents of the severe and highly contagious ocular infections acute hemorrhagic conjunctivitis and epidemic keratoconjunctivitis, respectively. Currently, neither vaccines nor antiviral agents are available for treating these diseases, which affect millions of individuals worldwide. CVA24v and HAdV-37 utilize sialic acid as attachment receptors facilitating entry into host cells. Previously, we and others have shown that derivatives based on sialic acid are effective in preventing HAdV-37 binding and infection of cells. Here, we designed and synthesized novel pentavalent sialic acid conjugates and studied their inhibitory effect against CVA24v and HAdV-37 binding and infection of human corneal epithelial cells. The pentavalent conjugates are the first reported inhibitors of CVA24v infection and proved efficient in blocking HAdV-37 binding. Taken together, the pentavalent conjugates presented here form a basis for the development of general inhibitors of these highly contagious ocular pathogens.

Microanatomy of the nerve to iliacus muscle and its clinical significance / Microanatomía del nervio del músculo ilíaco y su importancia clínica

The iliacus muscle, arising from iliac fossa is innervated chiefly by nerves to iliacus and femoral nerve. The tendon of iliacus muscle in the caudal part fuses with the tendon of psoas major muscle to form iliopsoas tendon As the iliacus/iliopsoas is responsible for flexing of the thigh and the forward tilting of the pelvis, body posture, Olympic lifts, daily activities like walking and running, so impairment of above functions, due to spinal cord injury or injury to nerves to iliacus, remained a grey area to explore manifestation of nerve lesions at fascicular level. Therefore an experimental study was designed to map the complex fascicular pathways suffering from splits, fusions and multiplexing coupled with measurement of distances of closely sampled histological slides. Tracking, correlation and interpretation of fascicles, in these slides of a cropped femoral nerve in iliacus region from a 70 year old female cadaver were analyzed. The study resulted in three schematic models of fascicular pathways in 3 nerves to iliacus and 2 tabular models of 2 remaining nerves to iliacus revealing complete picture of fascicles interrupted by dynamic transformational processes. These results would facilitate MRI neurographic interpretation at fascicular level and neurosurgical treatment through identification. The fascicular identification and setup would also discover anatomical complications and location of injury. Besides the huge data volume evolved off this experiment, the study would not only open up grey area for neuroanatomical research but also would revolutionize the neurosurgical repair and grafting of nerves to iliacus at fascicular level.

El músculo ilíaco, que se inserta en la fosa ilíaca, está inervado principalmente por los nervios ilíaco y femoral. El tendón del músculo ilíaco en la parte caudal se fusiona con el tendón del músculo psoas mayor para formar el tendón del músculo iliopsoas. Los músculos ilíaco e iliopsoas son responsables de la flexión del muslo y la inclinación hacia delante de la pelvis, la postura del cuerpo, los levantamientos olímpicos, las actividades diarias como caminar y correr, por lo que el deterioro de las funciones anteriores, debido a lesiones de la médula espinal o de los nervios ilíacos, constituyen una dificultad para explorar la manifestación de lesiones nerviosas a nivel fascicular. Por lo tanto, se diseñó un estudio experimental para mapear las complejas vías fasciculares que presentan divisiones, fusiones y multiplexación, junto con medición en muestras histológicas. Se analizó el seguimiento, correlación y la interpretación de los fascículos en muestras de secciones del nervio femoral en la región ilíaca de un cadáver femenino de 70 años. Se obtuvieron tres modelos esquemáticos de vías fasciculares en 3 ramos del nervio ilíaco y dos modelos tabulares de los 2 ramos nerviosos restantes del nervio ilíaco, que muestran una imagen completa de los fascículos interrumpidos por procesos de transformación dinámica. Estos resultados facilitarían la interpretación neurográfica de la resonancia nuclear magnética a nivel fascicular y el tratamiento neuroquirúrgico a través de su identificación. La identificación y configuración del fascículo también permitirían descubrir complicaciones anatómicas y la localización de la lesión. Además del enorme volumen de datos que se desprendió de este estudio, éste no solo contribuiría a la investigación neuroanatómica, sino también puede aportar a la reparación neuroquirúrgica y al injerto de nervios al músculo ilíaco a nivel fascicular.

Sialic Acid-Containing Glycans as Cellular Receptors for Ocular Human Adenoviruses: Implications for Tropism and Treatment.

Human adenoviruses (HAdV) are the most common cause of ocular infections. Species B human adenovirus type 3 (HAdV-B3) causes pharyngoconjunctival fever (PCF), whereas HAdV-D8, -D37, and -D64 cause epidemic keratoconjunctivitis (EKC). Recently, HAdV-D53, -D54, and -D56 emerged as new EKC-causing agents. HAdV-E4 is associated with both PCF and EKC. We have previously demonstrated that HAdV-D37 uses sialic acid (SA)-containing glycans as cellular receptors on human corneal epithelial (HCE) cells, and the virus interaction with SA is mediated by the knob domain of the viral fiber protein. Here, by means of cell-based assays and using neuraminidase (a SA-cleaving enzyme), we investigated whether ocular HAdVs other than HAdV-D37 also use SA-containing glycans as receptors on HCE cells. We found that HAdV-E4 and -D56 infect HCE cells independent of SAs, whereas HAdV-D53 and -D64 use SAs as cellular receptors. HAdV-D8 and -D54 fiber knobs also bound to cell-surface SAs. Surprisingly, HCE cells were found resistant to HAdV-B3 infection. We also demonstrated that the SA-based molecule i.e., ME0462, designed to bind to SA-binding sites on the HAdV-D37 fiber knob, efficiently prevents binding and infection of several EKC-causing HAdVs. Surface plasmon resonance analysis confirmed a direct interaction between ME0462 and fiber knobs. Altogether, we demonstrate that SA-containing glycans serve as receptors for multiple EKC-causing HAdVs, and, that SA-based compound function as a broad-spectrum antiviral against known and emerging EKC-causing HAdVs.

Decoy Receptor Interactions as Novel Drug Targets against EKC-Causing Human Adenovirus.

Epidemic keratoconjunctivitis (EKC) is a severe ocular disease and can lead to visual impairment. Human adenovirus type-37 (HAdV-D37) is one of the major causative agents of EKC and uses sialic acid (SA)-containing glycans as cellular receptors. Currently, there are no approved antivirals available for the treatment of EKC. Recently, we have reported that sulfated glycosaminoglycans (GAGs) bind to HAdV-D37 via the fiber knob (FK) domain of the viral fiber protein and function as decoy receptors. Based on this finding, we speculated that GAG-mimetics may act as artificial decoy receptors and inhibit HAdV-D37 infection. Repurposing of approved drugs to identify new antivirals has drawn great attention in recent years. Here, we report the antiviral effect of suramin, a WHO-approved drug and a widely known GAG-mimetic, against HAdV-D37. Commercially available suramin analogs also show antiviral effects against HAdV-D37. We demonstrate that suramin exerts its antiviral activity by inhibiting the attachment of HAdV-D37 to cells. We also reveal that the antiviral effect of suramin is HAdV species-specific. Collectively, in this proof of concept study, we demonstrate for the first time that virus binding to a decoy receptor constitutes a novel and an unexplored target for antiviral drug development.

Sulfated Glycosaminoglycans as Viral Decoy Receptors for Human Adenovirus Type 37.

Glycans on plasma membranes and in secretions play important roles in infection by many viruses. Species D human adenovirus type 37 (HAdV-D37) is a major cause of epidemic keratoconjunctivitis (EKC) and infects target cells by interacting with sialic acid (SA)-containing glycans via the fiber knob domain of the viral fiber protein. HAdV-D37 also interacts with sulfated glycosaminoglycans (GAGs), but the outcome of this interaction remains unknown. Here, we investigated the molecular requirements of HAdV-D37 fiber knob:GAG interactions using a GAG microarray and demonstrated that fiber knob interacts with a broad range of sulfated GAGs. These interactions were corroborated in cell-based assays and by surface plasmon resonance analysis. Removal of heparan sulfate (HS) and sulfate groups from human corneal epithelial (HCE) cells by heparinase III and sodium chlorate treatments, respectively, reduced HAdV-D37 binding to cells. Remarkably, removal of HS by heparinase III enhanced the virus infection. Our results suggest that interaction of HAdV-D37 with sulfated GAGs in secretions and on plasma membranes prevents/delays the virus binding to SA-containing receptors and inhibits subsequent infection. We also found abundant HS in the basement membrane of the human corneal epithelium, which may act as a barrier to sub-epithelial infection. Collectively, our findings provide novel insights into the role of GAGs as viral decoy receptors and highlight the therapeutic potential of GAGs and/or GAG-mimetics in HAdV-D37 infection.

Glycomics and Proteomics Approaches to Investigate Early Adenovirus-Host Cell Interactions.

Adenoviruses as most viruses rely on glycan and protein interactions to attach to and enter susceptible host cells. The Adenoviridae family comprises more than 80 human types and they differ in their attachment factor and receptor usage, which likely contributes to the diverse tropism of the different types. In the past years, methods to systematically identify glycan and protein interactions have advanced. In particular sensitivity, speed and coverage of mass spectrometric analyses allow for high-throughput identification of glycans and peptides separated by liquid chromatography. Also, developments in glycan microarray technologies have led to targeted, high-throughput screening and identification of glycan-based receptors. The mapping of cell surface interactions of the diverse adenovirus types has implications for cell, tissue, and species tropism as well as drug development. Here we review known adenovirus interactions with glycan- and protein-based receptors, as well as glycomics and proteomics strategies to identify yet elusive virus receptors and attachment factors. We finally discuss challenges, bottlenecks, and future research directions in the field of non-enveloped virus entry into host cells.

Generation and characterization of a novel candidate gene therapy and vaccination vector based on human species D adenovirus type 56.

The vectorization of rare human adenovirus (HAdV) types will widen our knowledge of this family and their interaction with cells, tissues and organs. In this study we focus on HAdV-56, a member of human Ad species D, and create ease-of-use cloning systems to generate recombinant HAdV-56 vectors carrying foreign genes. We present in vitro transduction profiles for HAdV-56 in direct comparison to the most commonly used HAdV-5-based vector. In vivo characterizations demonstrate that when it is delivered intravenously (i.v.) HAdV-56 mainly targets the spleen and, to a lesser extent, the lungs, whilst largely bypassing liver transduction in mice. HAdV-56 triggered robust inflammatory and cellular immune responses, with higher induction of IFNγ, TNFα, IL5, IL6, IP10, MCP1 and MIG1 compared to HAdV-5 following i.v. administration. We also investigated its potential as a vaccine vector candidate by performing prime immunizations in mice with HAdV-56 encoding luciferase (HAdV-56-Luc). Direct comparisons were made to HAdV-26, a highly potent human vaccine vector currently in phase II clinical trials. HAdV-56-Luc induced luciferase 'antigen'-specific IFNγ-producing cells and anti-HAdV-56 neutralizing antibodies in Balb/c mice, demonstrating a near identical profile to that of HAdV-26. Taken together, the data presented provides further insight into human Ad receptor/co-receptor usage, and the first report on HAdV-56 vectors and their potential for gene therapy and vaccine applications.

Triazole linker-based trivalent sialic acid inhibitors of adenovirus type 37 infection of human corneal epithelial cells.

Adenovirus type 37 (Ad37) is one of the principal agents responsible for epidemic keratoconjunctivitis (EKC), a severe ocular infection that remains without any available treatment. Recently, a trivalent sialic acid derivative (ME0322, Angew. Chem. Int. Ed., 2011, 50, 6519) was shown to function as a highly potent inhibitor of Ad37, efficiently preventing the attachment of the virion to the host cells and subsequent infection. Here, new trivalent sialic acid derivatives were designed, synthesized and their inhibitory properties against Ad37 infection of the human corneal epithelial cells were investigated. In comparison to ME0322, the best compound (17a) was found to be over three orders of magnitude more potent in a cell-attachment assay (IC50 = 1.4 nM) and about 140 times more potent in a cell-infection assay (IC50 = 2.9 nM). X-ray crystallographic analysis demonstrated a trivalent binding mode of all compounds to the Ad37 fiber knob. For the most potent compound ophthalmic toxicity in rabbits was investigated and it was concluded that repeated eye administration did not cause any adverse effects.

Adenovirus-based vaccines for fighting infectious diseases and cancer: progress in the field.

The field of adenovirology is undergoing rapid change in response to increasing appreciation of the potential advantages of adenoviruses as the basis for new vaccines and as vectors for gene and cancer therapy. Substantial knowledge and understanding of adenoviruses at a molecular level has made their manipulation for use as vaccines and therapeutics relatively straightforward in comparison with other viral vectors. In this review we summarize the structure and life cycle of the adenovirus and focus on the use of adenovirus-based vectors in vaccines against infectious diseases and cancers. Strategies to overcome the problem of preexisting antiadenovirus immunity, which can hamper the immunogenicity of adenovirus-based vaccines, are discussed. When armed with tumor-associated antigens, replication-deficient and oncolytic adenoviruses can efficiently activate an antitumor immune response. We present concepts on how to use adenoviruses as therapeutic cancer vaccines and consider some of the strategies used to further improve antitumor immune responses. Studies that explore the prospect of adenoviruses as vaccines against infectious diseases and cancer are underway, and here we give an overview of the latest developments.

Gastroprotective effect of aqueous extract and mucilage from Bryophyllum pinnatum (Lam.) Kurz.

CONTEXT: Bryophyllum pinnatum is used as traditional medicine in India, Africa, Tropical America and China for treatment of various diseases. B. pinnatum contains different groups of phytoconstituents viz., flavonoid, terpenoids, alkaloid, phenolic compounds. AIM: The present study was carried out to evaluate the gastroprotective activity of B. pinnatum whole plant aqueous extract, and mucilage (MUC) isolated from the whole plant against ethanol induced gastric ulcer. MATERIALS AND METHODS: Pretreatment of rats with aqueous extract at dose level of 500 and 750 mg/kg b.w., MUC at 500 mg/kg dose level and standard drug Rabeprazole at dose level of 20 mg/kg b.w. where given for 7 days. RESULTS: The aqueous whole plant extract of B. pinnatum at dose of 750 mg/kg p.o. and MUC at dose of 500 mg/kg p.o. markedly decrease the incidence of ulcers in ethanol induced ulcer rats. In ethanol induced ulcer rats, there was a decrease in the gastric volume, free and total acidity and ulcerative index as compared to the control group. Total carbohydrate content was found to be an increase as compare to control the group. The aqueous whole plant extract of B. pinnatum at dose of 750 mg/kg showed a significant reduction in the above parameters which was comparable to the standard drug rabeprazole (20 mg/kg). B. pinnatum extract and MUC showed protection index 72.69 and 69.65% respectively, whereas standard drug rabeprazole showed protection index 75.49%. CONCLUSIONS: Whole plant extracts of B. pinnatum and MUC has potent gastroprotective effect which can be further clinically studied for new drug development.

Deltamethrin induced toxicity and ameliorative effect of alpha-tocopherol in broilers.

Deltamethrin (DM) is a broad-spectrum insecticide mainly used to protect crops, fruit and vegetables from pests such as mites, ants, weevils and beetles. Birds, animals and human beings living in same ecosystem are directly or indirectly at the risk of exposure to this insecticide leading to substantial decrease in growth. Thus we studied DM induced toxicity and ameliorative effects of alpha-tocopherol in broiler birds. DM was estimated in liver, breast and leg muscles of chickens feeding with only DM or DM with alpha-tocopherol daily for 42 days. Birds exposed to DM showed a dose dependent decrease in body weight on 5th, 6th and 7th weeks as compared to controls, and alpha-tocopherol partially restored the reduction in body weight. DM residue was found higher in liver as compared to breast and leg muscles.

Studies on seed production of mungbean (Vigna radiata) sown at different dates.

To get maximum yield of kharif mungbean, a field experiment was conducted in a split plot design and replicated four times. Five varieties of mungbean viz., Pusa 105, Ganga 1, ML 682, PMB 14 and Pant Mung 4 were sown normal and late. Data on growth, development and yield components were recorded from ten randomly selected plants from each plots. Grain and straw yield were taken from the net harvested plot and computed to get kg ha(-1). Among the varieties, PMB 14 ranked first in terms of grain yield (549 kg ha(-1)) followed by Ganga 1 (521kg ha(-1)), ML 682(495 kg ha(-1)), Pant Mung 4 (475 kg ha(-1)) and Pusa 105 (438 kg ha(-1)). Highest grain yield (516 kg ha(-1)) was obtained in case of normal sowing whereas comparably lower yield (476 kg ha(-1)) was recorded under late sown condition. Significant interaction was noticed only in case of pods per plant. To obtain higher grain yield, mungbean should be sown at normal time (1st week of August) with genotype PMB 14.Most consistent performer under normal and delayed sowing was ML682. Pant Mung 4 was found to be most sensitive in delay planting than other tested varieties.

Effect of Asteracantha longifolia Nees. against CCl4 induced liver dysfunction in rat.

Significant recovery after treatment with the whole plant slurry of A.longifolia Nees. was observed in plasma AST, ALT and cholesterol levels in CCl4 induced hepatotoxic rats. This was amply supported by electron micrographs, which indicated normalization of cytoarchitecture of mitochondria and endoplasmic reticulum. The results suggest that the slurry of the plant is useful as a liver tonic.

Antimicrobial activity of crude extracts from plant parts and corresponding calli of Bixa orellana L.

Ethanol extracts from the different parts of B. orellana showed differential antimicrobial activity. It was found that the extracts of in vitro leaves showed maximum activity against Bacillus pumilus followed by the extracts from the roots and hypocotyls. The callus derived from different explants too showed antimicrobial activity. The leaf callus showed maximum activity. The zone of inhibition for the diluted extracts of in vitro hypocotyls and roots and their corresponding calli showed minimum zone of inhibition at concentration 24 mg/ml, whereas the diluted extract of in vitro leaves and leaf derived callus showed minimum zone of inhibition at 16 mg/ml.