Prevalence and identification of Candida sp. in pregnant women using VITEK-2

Background: Candida sp. is seen in several areas of body such as, mouth, groin area including vagina and digestive tract as thrush or gastroenteritis. The slide-culture technique and the VITEK-2 automated system were used for species-identification of the fungus; nonetheless, a gold standard or any first identification method would have inherent errors in arriving at a correct identification of a microorganism at species level.Methods: Morphological fungal criteria were ascertained with germ tubes, glucose agar, sugar fermentation and sugar assimilation tests Candida from vaginal swabs and other clinical samples of 85 infected pregnant women with diabetes, by growing swab lots on Sabouraud’s Dextrose Agar (SDA) plates, the slide culture technique and the VITEK-2 automated system.Results: Of 85 patients, 122 isolates in SDA culture were determined as 7 Candida sp.  with number of isolates of each species, as follows: 47 C. albicans, 9 C. famata, 11 C. glabrata, 13 C. guilliermondii, 8 C. krusei, 3 C. parapsilosis and 37 C. tropicalis from vaginal swabs. From 60 vaginal swabs, 46 urine samples and 12 throat swabs it was seen that C. albicans was most prevalent. However, withVITEK-2, 201 fungal strains were identified; Candida sp. was isolated in all samples: 59 C. albicans, 19 C. famata, 21 C. glabrata, 23 C. guilliermondii,18 C. krusei, 13 C. parapsilosis and 48 C. tropicalis.Conclusions: The most prevalent species among the isolated fungi was C. albicans, causing VC in diabetic pregnant women

Modulation of inhibitory activity of xylanase - α-amylase inhibitor protein (XAIP): binding studies and crystal structure determination of XAIP- II from Scadoxus multiflorus at 1.2 Å resolution

Background: Plants produce a wide range of proteinaceous inhibitors to protect themselves against hydrolytic enzymes. Recently a novel protein XAIP belonging to a new sub-family (GH18C) was reported to inhibit two structurally unrelated enzymes xylanase GH11 and α -amylase GH13. It was shown to inhibit xylanase GH11 with greater potency than that of α-amylase GH13. A new form of XAIP (XAIP-II) that inhibits α-amylase GH13 with a greater potency than that of XAIP and xylanase GH11 with a lower potency than that of XAIP, has been identified in the extracts of underground bulbs of Scadoxus multiflorus. This kind of occurrence of isoforms of inhibitor proteins is a rare observation and offers new opportunities for understanding the principles of protein engineering by nature. Results: In order to determine the structural basis of the enhanced potency of XAIP-II against α-amylase GH13 and its reduced potency against xylanase GH11 as compared to that of XAIP, we have purified XAIP-II to homogeneity and obtained its complete amino acid sequence using cloning procedure. It has been crystallized with 0.1 M ammonium sulphate as the precipitating agent and the three-dimensional structure has been determined at 1.2 Å resolution. The binding studies of XAIP-II with xylanase GH11 and α-amylase GH13 have been carried out with surface plasmon resonance (SPR). Conclusion: The structure determination revealed that XAIP-II adopts the well known TIM barrel fold. The xylanase GH11 binding site in XAIP-II is formed mainly with loop α3-β3 (residues, 102 - 118) which has acquired a stereochemically less favorable conformation for binding to xylanase GH11 because of the addition of an extra residue, Ala105 and due to replacements of two important residues, His106 and Asn109 by Thr107 and Ser110. On the other hand, the α-amylase binding site, which consists of α-helices α6 (residues, 193 - 206), α7 (residues, 230 - 243) and loop β6-α6 (residues, 180 - 192) adopts a stereochemically more favorable conformation due to replacements of residues, Ser190, Gly191 and Glu194 by Ala191, Ser192 and Ser195 respectively in α-helix α6, Glu231 and His236 by Thr232 and Ser237 respectively in α-helix α7. As a result, XAIP-II binds to xylanase GH11 less favorably while it interacts more strongly with α-amylase GH13 as compared to XAIP. These observations correlate well with the values of 4.2 × 10-6 M and 3.4 × 10-8 M for the dissociation constants of XAIP-II with xylanase GH11 and α-amylase GH13 respectively and those of 4.5 × 10-7 M and 3.6 × 10-6 M of XAIP with xylanase GH11 and α-amylase GH13 respectively

Polysaccharide binding sites in hyaluronate lyase-crystal structures of native phage-encoded hyaluronate lyase and its complexes with ascorbic acid and lactose

Hyaluronate lyases are a class of endoglycosaminidase enzymes with a high level of complexity and heterogeneity. The main function of the Streptococcus pyogenes bacteriophage protein hyaluronate lyase, HylP2, is to degrade hyaluronan into unsaturated disaccharide units. HylP2 was cloned, over-expressed and purified to homogeneity. The recombinant HylP2 exists as a homotrimer with a molecular mass of approximately 110 kDa under physiological conditions. The HylP2 was crystallized and the crystals were soaked in two separate reservoir solutions containing ascorbic acid and lactose, respectively. The crystal structures of native HylP2 and its two complexes with ascorbic acid and lactose have been determined. HylP2 folds into four distinct domains with a central core consisting of 16 antiparallel β-strands forming an irregular triangular tube designated as triple-stranded β-helix. The structures of complexes show that three molecules each of ascorbic acid and lactose bind to protein at the sugar binding groove in the triple-stranded β-helix domain. Both ascorbic acid and lactose molecules occupy almost identical subsites in the long saccharide binding groove. Both ligands are involved in several hydrogen bonded interactions at each subsite. The binding characteristics and stereochemical properties indicate that Tyr264 may be involved in the catalytic activity of HylP2. The mutation of Tyr264 to Phe264 supports this observation

Mobilization of Stem Cells Using G-CSF for Acute Ischemic Stroke: A Randomized Controlled, Pilot Study

Background. There is emerging evidence to support the use of granulocyte colony-stimulating factor (G-CSF) therapy in patients with acute ischemic stroke. Aims. To explore feasibility, safety, and preliminary efficacy of G-CSF therapy in patients with acute ischemic stroke. Patients and Method. In randomized study, 10 patients with acute ischemic stroke were recruited in 1 : 1 ratio to receive 10 μg/kg G-CSF treatment subcutaneously daily for five days with conventional care or conventional treatment alone. Efficacy outcome measures were assessed at baseline, one month, and after six months of treatment included Barthel Index (BI), National Institute of Health Stroke Scale, and modified Rankin Scale. Results. One patient in G-CSF therapy arm died due to raised intracranial pressure. No severe adverse effects were seen in rest of patients receiving G-CSF therapy arm or control arm. No statistically significant difference between intervention and control was observed in any of the scores though a trend of higher improvement of BI score is seen in the intervention group. Conclusion. Although this study did not have power to examine efficacy, it provides preliminary evidence of potential safety, feasibility, and tolerability of G-CSF therapy. Further studies need to be done on a large sample to confirm the results

Studies - An open randomized comparative study of oral itraconazole pulse and terbinafine pulse in the treatment of onychomycosis

BACKGROUND: Onychomycosis is a recalcitrant disease of the nails caused by dermatophytes, yeasts, and molds. AIMS: To compare the clinical efficacy of oral itraconazole pulse therapy and oral terbinafine pulse therapy in onychomycosis. METHODS: A randomized single-blind clinical comparative study was undertaken on 120 patients of onychomycosis during the period March 1999-February 2002. Sixty patients were randomly assigned to receive oral itraconazole 100 mg, two capsules twice daily for seven days a month and the other group of sixty patients received oral terbinafine 250 mg, one tablet twice daily for seven days every month. Four such monthly pulses were administered for each drug. The patients were evaluated at 4-weekly intervals till sixteen weeks and then at 24, 36 and 48 weeks. RESULTS: We observed a clinical cure rate of 82% and mycological cure rate of 90% in the group of patients treated with itraconazole while the group with terbinafine showed clinical and mycological cure rates of 79% and 87% respectively. This difference was not statistically significant. CONCLUSIONS: Both oral itraconazole and terbinafine are effective in the treatment of onychomycosis when administered in the pulse dosage form. Terbinafine is more cost effective while itraconazole has a broader spectrum of antimycotic activity

An Evaluation of Analgesic Activity of Leaf and Stem Bark Extracts of Ficus Religiosa in Wistar Rats

Introduction: Pain is an unpleasant sensory and emotional experience associated with actual or potential damage. Non- steroidal anti-inflammatory drug (NSAIDs) & Opioids are wide analgesics, but because of adverse effects, their use is limited. Ficus Religiosa is a traditional medicinal plant, its various parts have been used in treating some conditions. Aims & Objective: To evaluate the analgesic activity of methanolic extract of leaves and stem bark of Ficus Religiosa at different doses. Materials & Methodology: This study was conducted in the Department of Pharmacology. Two doses of methanolic extract of leaves (100 & 200 mg/kg) and stem bark (125 & 250 mg/kg) of ficus religiosa were used. Wistar rats were used to evaluate analgesic activity and it was evaluated by using analgesiometer by tail flick method. Phytochemical screening of both the extracts were also done. Results: Both the extracts at their respective doses showed significant analgesic activity as compared with the control group, whereas it was not comparable with the standard drug ibuprofen (40 mg/kg p.o). Conclusion: Therefore, we conclude that both the extracts of ficus religiosa were showing analgesic activity. But this analgesic activity is not comparable with the standard drug Ibuprofen