Quality control methods for fruit extracts of Kigelia africana using high performance thin layer chromatography.

Kigelia africana is a tree native to Africa but also found in eastern and southern parts of India with reported anti-bacterial, anti-inflammatory, and immunomodulatory activities. Verbascoside, caffeic acid and ferulic acid are important markers for the quality control of the plant. Two different HPTLC methods were developed and validated; method - 1 for estimation of verbascoside and caffeic acid while method - 2 for estimation of caffeic acid and ferulic acid. Developed methods were applied to the methanolic fruit extract to determine the quantities of markers. Both methods were found to be linear, specific, precise, accurate, sensitive and robust. Results indicated that both methods can be used for quantitative determination of verbascoside, caffeic acid and ferulic acid in fruit extract. The developed methods may be utilised as a part of the quality control and standardisation for the raw material and extracts of Kigelia africana and can also aid to chromatographic fingerprinting of the plant.

Efficacy and Safety of Directly Acting Antivirals in Patients with Hepatitis C Infection on Hemodialysis.

INTRODUCTION: The high prevalence of hepatitis C virus (HCV) infection among patients on maintenance hemodialysis (MHD) has been reported in India. Due to the strong association of HCV infection with death and cardiovascular disease, it is important to treat the infection. However, treatment poses a challenge since only a few directly acting antivirals recommended in the guidelines for HCV treatment in the dialysis population are available in India. Pangenotypic sofosbuvir has concerns about its safety due to its renal elimination. MATERIALS AND METHODS: This prospective study was undertaken between 2019 and 2020 among patients on hemodialysis with HCV infection. Clinical details, biochemical parameters, viral load, and genotyping were recorded and the outcome of treatment with sofosbuvir in combination with velpatasvir/daclatasvir for 12 weeks was noted. Descriptive and inferential statistical analysis was carried out. The Chi-squared/Fisher exact test was used. RESULTS: In the present study, 54 hemodialysis patients with HCV were treated with full doses of sofosbuvir and velpatasvir/daclatasvir. Genotype 1 was the most common, seen in 75.9% (n = 41). Around 96.29% (n = 52) of patients achieved sustained virological response (SVR) at the end of the study. None of the patients experienced serious side effects requiring dose reduction or discontinuation of the treatment. CONCLUSION: Sofosbuvir combination therapy offers an excellent response in dialysis patients irrespective of the genotype and presence of cirrhosis with minimal monitoring as in non-chronic kidney disease (CKD) patients.

Glycogen synthase kinase-3: A potential target for diabetes.

Elevated circulating glucose level due to ß-cell dysfunction has been a key marker of Type-II diabetes. Glycogen synthase kinase-3 (GSK-3) has been recognized as an enzyme involved in the control of glycogen metabolism. Consequently, inhibitors of GSK-3 have been explored for anti-diabetic effects in vitro and in animal models. Further, the mechanisms governing the regulation of this enzyme have been elucidated by means of a combination of structural and cellular biological investigations. This review article examines the structural analysis of GSK-3 as well as molecular modeling reports from numerous researchers in the context of the design and development of GSK-3 inhibitors. This article centers on the signaling pathway of GSK-3 relevant to its potential as a target for diabetes and discusses advancements till date on different molecular modification approaches used by researchers in the development of novel GSK-3 inhibitors as potential therapeutics for the treatment of Type II diabetes.

Clinical Significance of Myositis-Specific and Myositis-Associated Antibody Profiles in Dermatomyositis.

Background: Myositis-specific autoantibodies (MSA) and myositis-associated autoantibodies (MAA) are clinically useful biomarkers that point to the diagnosis, clinical manifestations, and prognosis of dermatomyositis (DM). Materials and Methods: To estimate the prevalence of MSA as well as MAA and analyze possible clinical correlations of these autoantibodies in patients diagnosed with DM, we conducted a cross-sectional study of 30 patients who were diagnosed with DM. Results: MSA were positive in 19 patients (63%) in which Mi 2 was positive in 8 (27%) patients, and this was the most frequently found MSA. A total of 11 (36.7%) patients showed positive MAA. AntiPM/Scl 75 and anti-Ro 52 were positive in 5 (16.7%) patients each and these were the most commonly found MAA. Anti-La was absent in all our patients. There were 8 (27%) patients in whom both MSA and MAA were positive. Either MSA and/or MAA were positive in 22 (73%) patients. On a bivariate analysis, the patients who were positive for anti-PM/Scl 75 showed a significant difference in manifesting cutaneous ulcers (P value 0.023). It was also found that anti-SAE-positive patients showed a significant difference with malignancy (P value 0.014). Anti-Ro 52-positive patients were less likely to have symmetrical proximal muscle weakness (P value 0.006). Conclusions: All patients who were anti-MDA 5 positive had myositis and none of the anti-MDA 5-positive patients had rapidly progressive interstitial lung disease (RPILD). More than one MSA in the same patient was noted in three patients.

A cross-sectional study of epidemiological determinants for Covid-19 infection among nurses of a tertiary care hospital.

BACKGROUND: All health care workers including nurses are working in the frontline against coronavirus disease 2019 (Covid-19), which keeps them at high risk of getting infected. This study was conducted to identify risk factors for Covid-19 infection and compliance to Covid appropriate behavior among nurses. MATERIAL AND METHODS: A cross-sectional study was conducted on 150 nurses in a tertiary care hospital attached to a medical college in Mumbai, from April 2020 to December 2020. Data were collected telephonically using an interviewer-administered pre-validated, semi-structured questionnaire. Data entry and analysis were performed using SPSS version 21.0. RESULTS: The mean age of the nurses was 38.19 ± 12.14 years. The majority (80.7%) were exposed to Covid-19 while taking active care of Covid patients; a total of 108 (72%) were symptomatic at the time of testing; dietary modifications because of fear of Covid were performed by 121 (80.2%); 92.77% used the appropriate personal protective equipment (PPE) category according to the workplace; 121 (80.77%) followed all steps of donning and doffing at all times, and 19 (12.77%) reported a breach in PPE. A greater proportion of nurses working in Covid duties opted for hospital isolation than home isolation (p = 0.003). Risk factors such as sleep, shift duty, shift pattern, food timing, mode of travel, and type of PPE during travel were also found to be significantly associated with work type - Covid versus non-Covid (p < 0.05). CONCLUSIONS: Use of workplace appropriate PPE, proper donning and doffing facilities, duty shifts with a fixed duration, adequate hand hygiene practices, and regular food intake with adequate sleep can prevent Covid-19 infection at the workplace among nurses.

Structure-based De Novo Design and Docking Studies of 5(S)-Methyl-L-Proline Containing Peptidomimetic Compounds as Dipeptidyl Peptidase-4 Inhibitors.

BACKGROUND: Diabetes affects millions of people worldwide, with predicted numbers of about 700 million adults affected by 2045. Among the several anti-diabetic drug therapies available in the market, Dipeptidyl Peptidase-4 (DPP-4) inhibitors have emerged as a promising therapeutic approach with scope for exploration in the segment of peptidomimetics. OBJECTIVE: Series of proline-containing peptidomimetic compounds were designed and investigated for their drug-likeness through Lipinski's rule of five, lead-likeness through the rule of three, predictive pharmacokinetic studies (absorption, distribution, metabolism, and excretion), and toxicity properties through in-silico approaches. The designed compounds were evaluated for their interactions with binding sites of the enzyme DPP-4 using an extra precision docking approach. METHODS: Proline-containing peptidomimetic compounds were designed rationally. Drug-likeness and lead-likeness properties were calculated using Schrödinger Maestro v11.2 software. ADME and toxicity properties were predicted using PreADMET version 2.0. Docking study was performed using Schrödinger Maestro v11.2 software, and ligands for the study were designed using MarvinSketch software. RESULTS: 5(S)-methyl-L-proline containing 17 ligands were designed. All of them were found to obey Lipinski's rule of five. Compounds were found to have good ADME profile and low toxicity predictions. CONCLUSION: Four compounds were found to have good interactions with DPP-4 binding sites and hence created the scope to develop DPP-4 inhibitors containing 5(S)-methyl-L-proline moiety.

Epidemiological determinants of leptospirosis in rural and urban districts of Maharashtra, India.

BACKGROUND: Leptospirosis is emerging as one of the growing public health problems in many parts of India. It can occur in both rural and urban areas with varied risk factors. This study was taken up in three districts of Maharashtra namely-Mumbai, Ratnagiri, and Sindhudurg to understand the determinants of leptospirosis in both the urban and rural areas and look for differences if any. MATERIALS AND METHODS: This cross-sectional study was carried out during the year 2017. A pretested validated questionnaire was used to collect data. Field observations were made. Eighty-seven cases from Sindhudurg and 14 from Ratnagiri and 307 cases from Mumbai were included in the study. RESULTS: A total of 408 cases were included in the study. A total of 63 (62.4%) were males and 38 (37.6%) were females. Most cases belonged to the 20-35 year age group (37%). In rural areas, 32.7% of them visited government facilities first, whereas, in the urban areas, it was 73.9% (P = 0.006). Headache, myalgia, and prostration were more common in cases from rural areas (P = <0.05). Skin rash was found to be associated with urban cases of leptospirosis. The presence of rodents, cattle sheds, pets, and working in paddy fields were common environmental risks in rural areas, and using water for recreational activities were common in urban areas (P < 0.001). CONCLUSIONS: Context-specific risk factors were found significantly associated with the cases. No important difference was found in the epidemiology of leptospirosis in the urban and rural areas except the source of infection.

(RE) currences in COVID-19: (RE)-activation or (RE)-infection?

The COVID-19 which started in December 2019 has spread rapidly worldwide with millions of cases out of which many have successfully recovered. Owing to novelty, COVID-19 has been a mystery in itself since the beginning. Some cases develop severe complications while some remain asymptomatic. With research being done in all parts of the world, many lacunae in the etiology and pathophysiology of the disease have been filled. As we move ahead with the pandemic new information is getting added to the existing knowledge pool. The residual damages and long-term health effects are yet to be encountered. The question that has been in the minds of many researchers and is still being explored remains that about re-infection. There are multiple cases worldwide where the discharged patients have been detected positive once again. In India, the guidelines for prophylaxis, testing strategy, quarantine, home isolation, and discharge policies have been revised time and again by the ministry and ICMR. It is difficult to label ones re-detected positive status, taking into consideration strain of coronavirus, dead viral particles, antibodies and reliability of tests. The role of vaccine and herd immunity also becomes controversial with number of such cases arising. We have tried to compile and find out the scientific causes, its effects on individual and what can be the implications on the Public Health and scope of development of strategies required for such occurrences. Efforts need to be taken in such a way that there is neither a panic situation nor should there be a false sense of security post-recovery.

Bringing Real-World Microbiology Experiences to Undergraduate Students in Resource-Limited Environments.

Undergraduate microbiology curriculum should be amenable to periodic changes to incorporate new developments and ideas. The curriculum should be used not merely as a way to disseminate facts but also as a way to allow students to experience the process of science. In the context of undergraduate microbiology education in Osmania University (Hyderabad, India), existing curriculum does not explicitly allow students to engage in deeper understanding of concepts and understanding of the process of science, both in lecture and laboratory courses. The assessment methods that are currently used are limited in scope as they only test factual recall and superficial understanding of the subject and very minimally assess critical thinking skills. Another factor hampering innovation in the broader context of undergraduate education is the unavailability and inaccessibility to adequate resources. To address the issue of resource-limitations in implementing activities that expose undergraduate students to real-world microbiology experiences, a collaboration between a research institute and two teaching colleges was formed. This collaboration involved teacher and student workshops on exploring microbial diversity using 16S rRNA analysis with a view of blending novel research questions with technical skills in the undergraduate microbiology lab. This effort is an example of educators providing students with authentic experiences and, helping them gain critical knowledge and research skills in microbiology even under resource constraints, and students demonstrating motivation to participate in similar activities in the future. The collaborative effort described here can be a broadly sustainable model to improve overall undergraduate education in relatively resource-limited environments.

Differences in serum markers of oxidative stress in well controlled and poorly controlled asthma in Sri Lankan children: a pilot study.

BACKGROUND: Asthma is a disease characterised by hyper responsiveness and bronchoconstriction of airways, and is a major health burden globally. A dysfunction of the oxidant-antioxidant balance, termed oxidative stress, has been implicated in the pathophysiology of asthma. The present study aims to assess the changes in oxidative stress markers, namely nitric oxide metabolites and antioxidant capacity, in children with poorly controlled and well controlled asthma, in comparison to healthy controls. METHODS: The present study enrolled 72 children (ages 5-15 years) classified into three groups: (1) poorly controlled asthma (n = 20), (2) well controlled asthma (n = 24) and (3) healthy controls (n = 27). An interviewer-administered questionnaire was used to record socio-demographic data of the participants. The serum concentrations of the oxidant markers (nitrite, nitrate and total nitric oxide metabolites [NOx]) were determined using the Griess test, and the total antioxidant capacity (TAOC) was determined using the ABTS decolorisation method. The concentrations of these markers were compared across the three groups. RESULTS: The three study groups were similar in terms of socio-demographic data. The differences across the three groups were statistically significant for serum concentrations of nitrate and NOx (but not nitrite) and serum TAOC. Further analyses showed that the disparity for nitrate and NOx concentrations was greatest between poorly controlled asthma and healthy controls (p = 0.001 and p < 0.001) compared to the well-controlled asthmatics and healthy controls (p = 0.036 and p = 0.049). A significant difference in serum nitrate and NOx concentrations was not observed between the two asthma groups (p = 0.311 and 0.203). The TAOC were significantly lower in poorly controlled asthmatics as compared to well-controlled asthmatics (p = 0.003) and healthy controls (p < 0.001). However, there was no significant difference in the serum TAOC between healthy controls and well-controlled asthmatics (p = 0.496). These findings may indicate that it is perhaps the higher TAOC that contributes to the well controlled state of asthma. CONCLUSIONS: The present study indicated that an imbalance of oxidants and antioxidants in the serum may have an underlying role in asthma pathophysiology, and how these markers may be effective in asthma management.

CoMFA, CoMSIA and HQSAR Analysis of 3-aryl-3-ethoxypropanoic Acid Derivatives as GPR40 Modulators.

BACKGROUND: Human GPR40 receptor, also known as free fatty-acid receptor 1, is a Gprotein- coupled receptor that binds long chain free fatty acids to enhance glucose-dependent insulin secretion. In order to improve the resistance and efficacy, computational tools were applied to a series of 3-aryl-3-ethoxypropanoic acid derivatives. A relationship between the structure and biological activity of these compounds, was derived using a three-dimensional quantitative structure-activity relationship (3D-QSAR) study using CoMFA, CoMSIA and two-dimensional QSAR study using HQSAR methods. METHODS: Building the 3D-QSAR models, CoMFA, CoMSIA and HQSAR were performed using Sybyl-X software. The ratio of training to test set was kept 70:30. For the generation of 3D-QSAR model three different alignments were used namely, distill, pharmacophore and docking based alignments. Molecular docking studies were carried out on designed molecules using the same software. RESULTS: Among all the three methods used, Distill alignment was found to be reliable and predictive with good statistical results. The results obtained from CoMFA analysis q2, r2cv and r2 pred were 0.693, 0.69 and 0.992 respectively and in CoMSIA analysis q2, r2cv and r2pred were 0.668, 0.648 and 0.990. Contour maps of CoMFA (lipophilic and electrostatic), CoMSIA (lipophilic, electrostatic, hydrophobic, and donor) and HQSAR (positive & negative contribution) provided significant insights i.e. favoured and disfavoured regions or positive & negative contributing fragments with R1 and R2 substitutions, which gave hints for the modifications required to design new molecules with improved biological activity. CONCLUSION: 3D-QSAR techniques were applied for the first time on the series 3-aryl-3- ethoxypropanoic acids. All the models (CoMFA, CoMSIA and HQSAR) were found to be satisfactory according to the statistical parameters. Therefore such a methodology, whereby maximum structural information (from ligand and biological target) is explored, gives maximum insights into the plausible protein-ligand interactions and is more likely to provide potential lead candidates has been exemplified from this study.

Combiphore (Structure and Ligand Based Pharmacophore) - Approach for the Design of GPR40 Modulators in the Management of Diabetes.

BACKGROUND: Pharmacophore mapping and molecular docking can be synergistically integrated to improve the drug design and discovery process. A rational strategy, combiphore approach, derived from the combined study of Structure and Ligand based pharmacophore has been described to identify novel GPR40 modulators. METHODS: DISCOtech module from Discovery studio was used for the generation of the Structure and Ligand based pharmacophore models which gave hydrophobic aromatic, ring aromatic and negative ionizable as essential pharmacophoric features. The generated models were validated by screening active and inactive datasets, GH scoring and ROC curve analysis. The best model was exposed as a 3D query to screen the hits from databases like GLASS (GPCR-Ligand Association), GPCR SARfari and Mini-Maybridge. Various filters were applied to retrieve the hit molecules having good drug-like properties. A known protein structure of hGPR40 (pdb: 4PHU) having TAK-875 as ligand complex was used to perform the molecular docking studies; using SYBYL-X 1.2 software. RESULTS AND CONCLUSION: Clustering both the models gave RMSD of 0.89. Therefore, the present approach explored the maximum features by combining both ligand and structure based pharmacophore models. A common structural motif as identified in combiphore for GPR40 modulation consists of the para-substituted phenyl propionic acid scaffold. Therefore, the combiphore approach, whereby maximum structural information (from both ligand and biological protein) is explored, gives maximum insights into the plausible protein-ligand interactions and provides potential lead candidates as exemplified in this study.

Giant Perivascular Epithelioid Cell Tumor of the Orbit: A Clinicopathological Analysis and Review of the Literature.

AIM: To describe and review the clinical, radiological, and histopathological characteristics of an orbital perivascular epithelioid cell tumor (PEComa). METHODS: A systematic review of clinical records, radiological investigations, microscopic features, and immunohistochemical characteristics was done. RESULTS: A 9-year-old female child presented with a year-long history of a large orbital mass associated with painless, progressive proptosis of the right eye. Radiologically, a well-defined orbital mass was seen with no intracranial extension. Excision was performed and histopathological examination showed uniform epithelioid cells in nests separated by thin fibrovascular septae. The tumor cells stained positively for Human Melanoma Black-45, but neg-atively for desmin, S-100, smooth muscle actin, MyoD1, microphthalmia-associated transcription factor, vimentin, CD10, CD31, and CD34 with a low proliferation index of 5-7%. Based on the tumor's morphological and immuno-histochemical characteristics, a diagnosis of giant orbital PEComa was made. No recurrence was seen at the last follow-up. CONCLUSIONS: PEComas are uncommon mesenchymal neoplasms that have typical histological features, with an immunohistochemical profile of negativity for epithelial markers and positivity for melanocytic markers. For benign PEComas, complete excision is advised. However, since PEComas elsewhere in the body have been known to be malignant, a close follow-up of such cases is recommended.

Celecoxib Enhances the Efficacy of Low-Dose Antibiotic Treatment against Polymicrobial Sepsis in Mice and Clinical Isolates of ESKAPE Pathogens.

Treatment of multidrug resistant bacterial infections has been a great challenge globally. Previous studies including our study have highlighted the use of celecoxib, a non-steroidal anti-inflammatory drug in combination with antibiotic has decreased the minimal inhibitory concentration to limit Staphylococcus aureus infection. However, the efficacy of this combinatorial treatment against various pathogenic bacteria is not determined. Therefore, we have evaluated the potential use of celecoxib in combination with low doses of antibiotic in limiting Gram-positive and Gram-negative bacteria in vivo in murine polymicrobial sepsis developed by cecum ligation and puncture (CLP) method and against clinically isolated human ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species). The in vivo results clearly demonstrated a significant reduction in the bacterial load in different organs and in the inflammatory markers such as COX-2 and NF-κB via activation of SIRT1 in mice treated with imipenem, a choice of antibiotic for polymicrobial sepsis treatment. Combinatorial treatment of ampicillin and celecoxib was effective on clinical isolates of ESKAPE pathogens, 45% of tested clinical isolates showed more than 50% reduction in the colony forming units when compared to ampicillin alone. In conclusion, this non-traditional treatment strategy might be effective in clinic to reduce the dose of antibiotic to treat drug-resistant bacterial infections.

Levonorgestrel intrauterine system (Mirena): An emerging tool for conservative treatment of abnormal uterine bleeding.

INTRODUCTION: To study the efficacy of levonorgestrel intrauterine system (LNG-IUS; Mirena) in conservative management of abnormal uterine bleeding (AUB). MATERIALS AND METHODS: Seventy women between 30 and 55 years with AUB were included in a study conducted over a period of 3 years. Response was assessed monthly for first 4 months and then yearly for maximum 2 years. RESULTS: Mirena caused a 80% decrease in median menstrual blood loss (MBL) at 4 months, 95% decrease in MBL by 1 year, and 100% decrease (amenorrhea) by 2 years. Mean hemoglobin (Hb) % showed a significant rise of 7.8% from baseline 4 months post Mirena insertion. Mirena acted as an effective contraceptive in women not using any other form of contraception. Hysterectomy could be avoided in most of the women. CONCLUSION: Mirena provides an incredible nonsurgical alternative in treatment of menorrhagia. Its effects are reversible and it is an excellent fertility-sparing device. It is also an effective contraceptive.

DOPA and DHN pathway orchestrate melanin synthesis in Aspergillus species.

Melanins are high molecular weight hydrophobic pigments that have been studied for their role in the virulence of fungal pathogens. We investigated the amount and type of melanin in 20 isolates of Aspergillus spp.; A. niger (n = 3), A. flavus (n = 5), A. tamarii (n = 3), A. terreus (n = 3), A. tubingensis (n = 3), A. sydowii (n = 3). Aspergillus spp. were identified by sequencing the internal transcribed spacer (ITS) region. Extraction of melanin from culture filtrate and fungal biomass was done and followed by qualitative and quantitative analysis of melanin pigment. Ultraviolet (UV), Fourier transformed infrared (FT-IR), and electron paramagnetic resonance (EPR) spectra analyses confirmed the presence of melanin. The melanin pathway was studied by analyzing the effects of inhibitors; kojic acid, tropolone, phthalide, and tricyclazole. The results indicate that in A. niger and A. tubingensis melanin was found in both culture filtrate and fungal biomass. For A. tamarii and A. flavus melanin was extracted from biomass only, whereas melanin was found only in culture filtrate for A. terreus. A negligible amount of melanin was found in A. sydowii. The maximum amount of melanin from culture filtrate and fungal biomass was found in A. niger and A. tamarrii, respectively. The DOPA (3,4-dihydroxyphenylalanine) pathway produces melanin in A. niger, A. tamarii and A. flavus, whereas the DHN (1,8-dihydroxynaphthalene) pathway produces melanin in A. tubingensis and A. terreus. It can be concluded that the amount and type of melanin in aspergilli largely differ from species to species.

Microscopic evaluation, molecular identification, antifungal susceptibility, and clinical outcomes in fusarium, Aspergillus and, dematiaceous keratitis.

PURPOSE: Fusarium, Aspergillus, and Dematiaceous are the most common fungal species causing keratitis in tropical countries. Herein we report a prospective study on fungal keratitis caused by these three fungal species. METHODOLOGY: A prospective investigation was undertaken to evaluate eyes with presumed fungal keratitis. All the fungal isolates (n = 73) obtained from keratitis infections were identified using morphological and microscopic characters. Molecular identification using sequencing of the ITS region and antifungal susceptibility tests using microdilution method were done. The final clinical outcome was evaluated in terms of the time taken for resolution of keratitis and the final visual outcome. The results were analyzed after segregating the cases into three groups, namely, Fusarium, Aspergillus, and Dematiaceous keratitis. RESULTS: Diagnosis of fungal keratitis was established in 73 (35.9%) cases out of 208 cases. The spectra of fungi isolated were Fusarium spp. (26.6%), Aspergillus spp. (21.6%), and Dematiaceous fungi (11.6%). The sequence of the ITS region could identify the Fusarium and Aspergillus species at the species complex level, and the Dematiaceous isolates were accurately identified. Using antifungal agents such as fluconazole, natamycin, amphotericin B, and itraconazole, the minimum inhibitory concentrations (MICs) for Fusarium spp. were >32 µ g/mL, 4-8 µ g/mL, 0.5-1 µ g/mL, and >32 µ g/mL, respectively. Antifungal susceptibility data showed that Curvularia spp. was highly resistant to all the antifungal agents. Overall, natamycin and amphotericin B were found to be the most effective antifungal agents. The comparative clinical outcomes in all cases showed that the healing response in terms of visual acuity of the Dematiaceous group was significantly good when compared with the Fusarium and Aspergillus groups (P < 0.05). The time required for healing in the Fusarium group was statistically significantly less when compared with the Aspergillus and Dematiaceous groups. CONCLUSION: This study demonstrates important differences in microscopic features of scraping material and antifungal susceptibility between the three groups. Early and accurate identification coupled with the MIC data, and thereby appropriate treatment is crucial for complete recovery.

Estrogen mediated protection of cytoskeleton against oxidative stress.

BACKGROUND & OBJECTIVES: Cytoskeletal proteins are deregulated during oxidative stress and cataract formation. However, estrogen which protects against cataract formation and harmful effects of oxidative stress has not been tested on the cytoskeleton of lens epithelial cells (LECs). The current study was undertaken to assess if the protection rendered to LECs by estrogen was mediated by preserving the cytoskeletal proteins. METHODS: Oxidative stress was induced by 50 µM of H 2 O 2 in cultured goat LECs (gLECs) and effect of 1 µM 17ß-estradiol (E 2 ) was tested. After treatment, morphological analysis of cells was carried out using haematoxylin-eosin staining and cell density was also quantified. Cell viability was determined using Hoechst (Ho), YO-Pro (YP) and propidium iodide (PI). F-actin and vimentin were localized using phalloidin and anti-vimentin antibody, respectively, and viewed under fluorescence microscopy. Vimentin was further analysed at protein level by Western blotting. RESULTS: H 2 O 2 led to increased condensation of nucleus, cell death and apoptosis but these were prevented with pre- and co-treatment of E 2 with increase in cell viability (P<0.001). E 2 also prevented H 2 O 2 mediated depolymerization of cytoskeleton but was not able to reverse the changes when given after induction of oxidative stress. INTERPRETATION & CONCLUSIONS: Our findings showed that E 2 helped in preventing deteriorating effect of H 2 O 2 , inhibited cell death, apoptosis and depolymerisation of cytoskeletal proteins in LECs. However, the exact mechanism by which estrogen renders this protection to cytoskeleton of lens epithelial cells remains to be determined.

Redox regulation of T-cell function: from molecular mechanisms to significance in human health and disease.

Reactive oxygen species (ROS) are thought to have effects on T-cell function and proliferation. Low concentrations of ROS in T cells are a prerequisite for cell survival, and increased ROS accumulation can lead to apoptosis/necrosis. The cellular redox state of a T cell can also affect T-cell receptor signaling, skewing the immune response. Various T-cell subsets have different redox statuses, and this differential ROS susceptibility could modulate the outcome of an immune response in various disease states. Recent advances in T-cell redox signaling reveal that ROS modulate signaling cascades such as the mitogen-activated protein kinase, phosphoinositide 3-kinase (PI3K)/AKT, and JAK/STAT pathways. Also, tumor microenvironments, chronic T-cell stimulation leading to replicative senescence, gender, and age affect T-cell susceptibility to ROS, thereby contributing to diverse immune outcomes. Antioxidants such as glutathione, thioredoxin, superoxide dismutase, and catalase balance cellular oxidative stress. T-cell redox states are also regulated by expression of various vitamins and dietary compounds. Changes in T-cell redox regulation may affect the pathogenesis of various human diseases. Many strategies to control oxidative stress have been employed for various diseases, including the use of active antioxidants from dietary products and pharmacologic or genetic engineering of antioxidant genes in T cells. Here, we discuss the existence of a complex web of molecules/factors that exogenously or endogenously affect oxidants, and we relate these molecules to potential therapeutics.

Cx43, ZO-1, alpha-catenin and beta-catenin in cataractous lens epithelial cells.

Specimens of the anterior lens capsule with an attached monolayer of lens epithelial cells (LECs) were obtained from patients (n=52) undergoing cataract surgery. Specimens were divided into three groups based on the type of cataract: nuclear cataract, cortical cataract and posterior subcapsular cataract (PSC). Clear lenses (n=11) obtained from donor eyes were used as controls. Expression was studied by immunofluorescence, real-time PCR and Western blot. Statistical analysis was done using the student's t-test. Immunofluorescence results showed punctate localization of Cx43 at the cell boundaries in controls, nuclear cataract and PSC groups. In the cortical cataract group, cytoplasmic pools of Cx43 without any localization at the cell boundaries were observed. Real-time PCR results showed significant up-regulation of Cx43 in nuclear and cortical cataract groups. Western blot results revealed significant increase in protein levels of Cx43 and significant decrease of ZO-1 in all three cataract groups. Protein levels of alpha-catenin were decreased significantly in nuclear and cortical cataract group. There was no significant change in expression of beta-catenin in the cataractous groups. Our findings suggest that ZO-1 and alpha-catenin are important for gap junctions containing Cx43 in the LECs. Alterations in cell junction proteins may play a role during formation of different types of cataract.