Soil polluted with nano ZnO reveals unstable bacterial communities and decoupling of taxonomic and functional diversities.

Due to relentless production and disposal of nano zinc oxide (nZnO), it has become critical to comprehend the serious risks large-scale accumulation of nZnO pose to bacterial communities in soil. The primary objective was to evaluate the changes in bacterial community structure and associated functional pathways through predictive metagenomic profiling and subsequent validation through Quantitative Realtime PCR in soil spiked with nZnO (0, 50, 200, 500 and 1000 mg Zn kg-1) and similar levels of bulk ZnO (bZnO). The results revealed that soil microbial biomass-C, -N, -P, soil respiration and enzyme activities decreased markedly at higher ZnO levels. The alpha diversity decreased with increasing ZnO level, with more impact under nZnO, while beta diversity analyses indicated a distinct dose- dependent separation of bacterial communities. The dominant taxa including Proteobacteria, Bacterioidetes, Acidobacteria and Planctomycetes significantly increased in abundance, while Firmicutes, Actinobacteria and Chloroflexi decreased in abundance with elevated nZnO and bZnO levels. Redundancy analysis indicated that changes in bacterial community structure instilled a greater dose- rather than size- specific response on key microbial parameters. Predicted key functions did not show a dose- specific response, and at 1000 mg Zn kg-1, methane metabolism as well as starch and sucrose metabolism were attenuated, while functions involving two component systems and bacterial secretion systems were enhanced under bZnO indicating better stress avoidance mechanism than under nZnO. Realtime PCR and microbial endpoint assays confirmed the metagenome derived taxonomic and functional data, respectively. Taxa and functions that varied substantially under stress were established as bioindicators to predict nZnO toxicity in soils. Taxon-function decoupling indicated that the soil bacterial communities deployed adaptive mechanisms under high ZnO, with lesser buffering capacity and resilience of communities under nZnO.

Understandability and Actionability of Available Video Information on YouTube Regarding Hemophilia: A Cross-Sectional Study.

Introduction With the advent of revolutionary information technology, most general medical information can be accessed by the community at large. However, the factual nature of information, its understandability, and actionability of diseases like Hemophilia are unknown to the general population. Hence the present study has been envisaged to assess the understandability and actionability of available video information on YouTube about Hemophilia. Methods A cross-sectional study was performed using the Patient Education Materials Assessment Tool for Audiovisual materials (PEMAT-AV) to assess the understandability and actionability of 50 videos shown by order of relevance utilizing three independent assessors. An online google survey was prepared using the PEMAT questionnaire as a basis and results were recorded and saved as a Microsoft Excel sheet for analysis. Data was analyzed using either Microsoft Excel or an online calculator as the case may be. Results A total of 50 short videos on Hemophilia were assessed by three independent assessors using PEMAT. The data so obtained was rechecked by an independent reviewer before data analysis. Three videos were excluded due to non-English language while only two videos out of 50 showed 100% average understandability and actionability. Average understandability and actionability scores range between 34 to 100 percent and 11.1 to 100 percent, respectively. Most videos have higher average understandability than actionability (P value=0.003). Conclusion Our study shows there are only a few high-quality short videos available as audio-visual patient education materials on YouTube about Hemophilia. There is a great need to develop content that is beneficial to patients as patient educational material.

Osteoinduction Using a Prefabricated Scaffold in the Gluteus Muscle of Wistar Rats: An Animal Study.

Introduction: Bone grafts are very significant for the reconstruction of the trauma and the deformations created thereafter. Various bone forming and promoting agents are widely studied. Hence, in this study, we aim to evaluate the osteoinduction effect of scaffold impregnated with Simvastatin in wistor rats in an observational study. Material and Methods: We implanted 18 Wistar rats to be equally divided as controls and test groups. Polycaprolactone coated with 20 mg concentration of Simavastatin under sterile condition was used as test and hydroxy appetite as control. Then, we observed the histopathological Heamatoxilin and Eosin and ABH, sections at 4, 12, and 26 weeks. We noted the calcium deposition and bone formation. Results: We observed a significant variation among groups at 4, 12, and 26 weeks in the calcium deposition. While for the bone formation no significant variation were noted among groups at 4 and 12 weeks, however, there was a significant difference noted at 26 weeks. At the end of 12 weeks, mild fibroblast proliferation was seen in the surrounding area. After 26 weeks, ABH sections showed focal area of fibroblast proliferation with mild mononuclear infiltration was noticed; but implant could not be located. Conclusion: We conclude that Simavastatin showed an anabolic effect on bone healing.

Colour Doppler evaluation of uterine and ovarian blood flow in patients of polycystic ovarian disease and post-treatment changes.

AIM: To assess the morphology and colour Doppler parameters in patients with polycystic ovarian syndrome (PCOS) and also to assess the changes in Doppler parameters in follow-up patients, who underwent treatment. MATERIALS AND METHODS: The study was conducted on 50 women of reproductive age who had clinical and biochemical findings suggestive of PCOS. Clinico-hormonal parameters were recorded. Ultrasound and colour Doppler flow measurements of bilateral ovaries were performed in the early proliferative phase of the menstrual cycle. After assessment of the bilateral ovaries, colour Doppler ultrasound was used to evaluate the main uterine artery at the cervico-uterine junction. Follow-up imaging after 3 months was undertaken in patients who underwent treatment (metformin) and changes in the imaging and hormonal parameters were correlated. RESULTS: The mean value of luteinising hormone (LH) and the ratio of LH: follicle-stimulating hormone (FSH) was significantly higher in PCOS patients. Ultrasound parameters were significantly higher in PCOS patients. Ovarian stromal vessels in PCOS patients had a significantly higher peak systolic velocity (PSV), low resistance index (RI), and pulsatility index (PI). The PSV of uterine arteries were significantly decreased and the RI and PI were significantly increased. On follow-up patients revealed changes in hormonal parameters. CONCLUSION: PCOS is a heterogeneous disorder and is a convergence of multisystem endocrine derangements. Ultrasound is good diagnostic tool for PCOS and the use of Doppler aids in the evaluation of haemodynamic changes in small vessels of utero-ovarian circulation and in response assessment.

Interplay of genes in plant-pathogen interactions: In planta expression and docking studies of a beta 1,3 glucanase gene from Piper colubrinum and a glucanase inhibitor gene from Phytophthora capsici.

Oomycete pathogen, Phytophthora capsici is devastating for black pepper (Piper nigrum L.) and causes foot rot disease at all stages of plant growth. Phytophthora secretes a glucanase inhibitor protein (GIP), which is capable of inhibiting defence proteins like endoglucanases. In this particular study Quantitative PCR analysis, molecular docking studies and analysis of sequences of Glucanase inhibitor protein and beta-1,3 glucanse genes were done mainly depending on the data derived from Phytophthora capsici whole genome sequencing and Piper colubrinum RNA-sequencing (RNA-Seq). Amino acid sequence length of GIP gene from P. capsici was about 353 amino acids and that of glucanase pcEGase gene from P. colubrinum was about 312 amino acids. GIP gene from P. capsici showed high level of expression at early hours of the inoculation time period and pcEGase gene showed high level of expression at 16 hpi. High level of expression of pcEGase gene at 16 hpi is an indication that the GIP gene is successfully inhibited by the glucanase protein from the plant. Moreover insilico studies gave some hint on the importance of certain sites on the surfaces of both interacting proteins that might be having a role in binding of the two proteins and subsequent reactions thereof. Insilico analysis also conclusively proved that inhibition of glucanase inhibitor protein is mainly caused by recognition of an arginine as well as an isoleucine residue during the interaction of the two proteins.

AMP-acetyl CoA synthetase from Leishmania donovani: identification and functional analysis of 'PX4GK' motif.

An adenosine monophosphate forming acetyl CoA synthetase (AceCS) which is the key enzyme involved in the conversion of acetate to acetyl CoA has been identified from Leishmania donovani for the first time. Sequence analysis of L. donovani AceCS (LdAceCS) revealed the presence of a 'PX4GK' motif which is highly conserved throughout organisms with higher sequence identity (96%) to lower sequence identity (38%). A ∼ 77 kDa heterologous protein with C-terminal 6X His-tag was expressed in Escherichia coli. Expression of LdAceCS in promastigotes was confirmed by western blot and RT-PCR analysis. Immunolocalization studies revealed that it is a cytosolic protein. We also report the kinetic characterization of recombinant LdAceCS with acetate, adenosine 5'-triphosphate, coenzyme A and propionate as substrates. Site directed mutagenesis of residues in conserved PX4GK motif of LdAceCS was performed to gain insight into its potential role in substrate binding, catalysis and its role in maintaining structural integrity of the protein. P646A, G651A and K652R exhibited more than 90% loss in activity signifying its indispensible role in the enzyme activity. Substitution of other residues in this motif resulted in altered substrate specificity and catalysis. However, none of them had any role in modulation of the secondary structure of the protein except G651A mutant.

Intravitreal bevacizumab in the treatment of vasoproliferative retinal tumours.

AIM: To evaluate the efficacy of intravitreal bevacizumab in the treatment of retinal vasoproliferative tumours (VPT). MATERIALS AND METHODS: Six eyes of 6 patients with VPT who received intravitreal bevacizumab were retrospectively reviewed. All patients received between one and three injections of intravitreal bevacizumab depending upon response to treatment. Best-corrected visual acuity (BCVA), tumour size, and presence of co-pathology or sequelae were noted pre- and postoperatively and then analysed. Subsequent retreatments were performed in patients with recurrent or persistent VPT according to the ophthalmologist's discretion. Retreatments included photodynamic therapy with verteporfin, ruthenium-106 plaque brachytherapy, or endoresection of tumour. RESULTS: The mean follow-up duration was 33.3 months (range 10-66 months). At baseline, the mean logMAR BCVA was 1.45 (Snellen equivalent of 6/165); range 0.10-1.90 (6/8-CF). Following bevacizumab treatment the mean logMAR BCVA was 0.98 (Snellen equivalent of 6/57); range 0.5-1.9 (Snellen equivalent of 6/19 to CF). Therefore, there was no statistically significant change in visual acuity. The mean tumour thickness reduced from 2.4 to 2.1 mm following treatment with bevacizumab. However, this did not reach the statistical significance of P<0.05. Despite the visual improvement following bevacizumab therapy, five out of six patients had recurrence of tumour activity during the follow-up period and required further intervention in order to achieve sustained regression. CONCLUSIONS: Intravitreal bevacizumab appeared to result in temporary reduction of tumour thickness in 3 out of 6 VPT patients. However, neither the reduction in tumour thickness nor the change in visual acuity were statistically significant and intravitreal bevacizumab monotherapy had limited effectiveness in causing long-term regression of the lesions. Additional therapy was indicated in five out of six patients to establish long-term regression. The efficacy of bevacizumab as an adjunct is as yet undetermined and further studies are needed. Presently, we recommend other treatment modalities in the long-term management of VPTs.

Disulfiram targets cancer stem-like cells and reverses resistance and cross-resistance in acquired paclitaxel-resistant triple-negative breast cancer cells.

BACKGROUND: Triple-negative breast cancer (TNBC) has significantly worse prognosis. Acquired chemoresistance remains the major cause of therapeutic failure of TNBC. In clinic, the relapsed TNBC is commonly pan-resistant to various drugs with completely different resistant mechanisms. Investigation of the mechanisms and development of new drugs to target pan-chemoresistance will potentially improve the therapeutic outcomes of TNBC patients. METHODS: In this study, 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT), combination index (CI)-isobologram, western blot, ALDEFLUOR analysis, clonogenic assay and immunocytochemistry were used. RESULTS: The chemoresistant MDA-MB-231PAC10 cells are highly cross-resistant to paclitaxel (PAC), cisplatin (CDDP), docetaxel and doxorubicin. The MDA-MB-231PAC10 cells are quiescent with significantly longer doubling time (64.9 vs 31.7 h). This may be caused by high expression of p21(Waf1). The MDA-MB-231PAC10 cells express high aldehyde dehydrogenase (ALDH) activity and a panel of embryonic stem cell-related proteins, for example, Oct4, Sox2, Nanog and nuclealisation of HIF2α and NF-κBp65. We have previously reported that disulfiram (DS), an antialcoholism drug, targets cancer stem cells (CSCs) and enhances cytotoxicity of anticancer drugs. Disulfiram abolished CSC characters and completely reversed PAC and CDDP resistance in MDA-MB-231PAC10 cells. CONCLUSION: Cancer stem cells may be responsible for acquired pan-chemoresistance. As a drug used in clinic, DS may be repurposed as a CSC inhibitor to reverse the acquired pan-chemoresistance.

Identification and synthesis of impurities formed during sertindole preparation.

Sertindole (1), an atypical anti-psychotic drug is used for the treatment of schizophrenia. During the laboratory optimization and later during its bulk synthesis the formation of various impurities was observed. The impurities formed were monitored and their structures were tentatively assigned on the basis of their fragmentation patterns in LC-MS. Most of the impurities were synthesized and their assigned constitutions confirmed by co-injection in HPLC. We describe herein the formation, synthesis and characterization of these impurities. Our study will be of immense help to others to obtain chemically pure sertindole.

Comparative evaluation of 2.0-mm locking plate system vs 2.0-mm nonlocking plate system for mandibular fracture: a prospective randomized study.

This study evaluated the efficacy of a 2.0-mm locking plate/screw system compared with a 2.0-mm non-locking plate/screw system in mandibular fractures. A prospective randomized clinical trial was conducted. Patients were randomly assigned to receive 2.0-mm locking plates (group A) or 2.0-mm nonlocking plates (group B). All patients were followed up for 12 weeks postoperatively. Complications were analysed according to the type of plate used and the site of fracture. Fifty patients with 76 fractures met the inclusion criteria. Thirty-six fracture sites were treated with 2.0-mm locking plates and 40 with 2.0-mm nonlocking plates. The number of patients requiring postoperative maxillomandibular fixation was significantly higher in group B (p < 0.01); seven complications occurred representing 9% of the total. Two complications occurred in the locking group and five in the nonlocking group with complication rates equalling 6% and 13%, respectively. When comparing the overall complication rates according to plates used, the χ(2) test showed no statistically significant difference between the locking and nonlocking plates (p > 0.05). In conclusion, mandible fractures treated with 2.0-mm locking plates and 2.0-mm nonlocking plates present similar short-term complication rates.

Reconstruction of ankylosed temporomandibular joint: Sternoclavicular grafting as an approach to management.

A retrospective analysis of 15 cases of ankylosed temporomandibular joint (TMJ) reconstructed, with sternoclavicular joint graft (SCG), during the period 2002-2007 was undertaken. All cases were analyzed for functional adaptation of the graft, considering maximum interincisal opening, and protrusive and laterotrusive movement of the jaw. Significant improvement was noticed in all cases except one, although maximum improvement was seen 3-6 months postoperatively. Radiological evaluation was carried out at regular intervals for 2-3 years to assess the anatomical adaptation of the graft. No major postoperative complications were observed and all the cases showed complete regeneration of the clavicle during follow up. This finding indicates that reconstruction of ankylosed TMJ with sternoclavicular joint graft is a satisfactory method of treatment.

Operative performance in laparoscopic cholecystectomy using the Procedural-Based Assessment tool.

AIMS: The Intercollegiate Surgical Curriculum Project (ISCP) has devised assessment tools for index operations to assess trainee technical skills. In this study we used the Procedural-Based Assessment (PBA) tool to evaluate operations performed by trainees. METHODS: Live and simulated laparoscopic cholecystectomies were performed by trainees. Two experienced surgeons assessed each operation blindly and independently. RESULTS: Eighty-four live (supervised) and 112 simulated (unsupervised) operations were performed by 28 trainees. Mean inter-rater reliability was kappa = .86 and .84 for live and simulated operations, respectively. Construct validity using Mann-Whitney for generic technical skills was significant for live and simulated operations, P < or = .05. Assessing specific technical skills showed construct validity for simulated unsupervised operations only, Mann-Whitney P < .05, but not for supervised live operations, Mann-Whitney P > .05. CONCLUSIONS: The PBA showed good inter-rater reliability. Assessing generic technical skills, PBA showed construct validity for both types of operations and for specific technical skills in the unsupervised simulated operations. We conclude that the PBA seems to be a reliable and valid assessment tool for generic technical skills in unsupervised simulated and live supervised laparoscopic cholecystectomies.

Angiogenesis, vascular endothelial growth factor and its receptors in human surgical wounds.

BACKGROUND: Angiogenesis plays an essential role in tissue repair. Vascular endothelial growth factor (VEGF) mediates angiogenesis through receptor kinases VEGF-R1 and VEGF-R2, and co-receptors, neuropilins Np1 and Np2. This study examined the spatial and temporal expression of these factors in relation to angiogenesis in surgical wounds. METHODS: Scar biopsies were obtained from patients between 3 days and 2 years after surgery. Normal skin control biopsies were taken during surgery. Microvessel density (MVD) was quantified using a Chalkley grid. VEGF, VEGF-R1, VEGF-R2, Np1 and Np2 endothelial expression was determined by immunohistochemistry, and correlated with MVD and scar age. RESULTS: Cumulative MVD was significantly greater in scars than controls (P = 0.011), and was related to scar age (P = 0.007). Expression of VEGF, VEGF-R2, Np1 and Np2 was increased significantly in all scars and correlated with MVD. In contrast, scar VEGF-R1 expression was decreased, and correlated with increased VEGF and VEGF-R2. CONCLUSION: Levels of VEGF, VEGF-R2, Np1 and Np2 are increased, whereas VEGF-R1 expression is decreased in angiogenesis, suggesting a role for VEGF-receptor complexes in early wound healing. This altered protein expression and increased presence of vessels is prolonged, suggesting that structural remodelling continues for at least 2 years after surgery.

Rapid high-performance liquid chromatography-tandem mass spectrometry method for simultaneous measurement of venlafaxine and O-desmethylvenlafaxine in human plasma and its application in comparative bioavailability study.

A rapid high-performance liquid chromatography-tandem mass spectrometry method has been developed and validated for simultaneous measurement of venlafaxine and O-desmethylvenlafaxine in human plasma using fluoxetine as an internal standard. In the liquid-liquid extraction method, compounds and internal standard were extracted from plasma using methyl tertiary butyl ether as an extraction solvent. The HPLC separation of the analytes was performed on a Zorbax SB-C(18), 50 x 4.6 mm, 5 microm column, using a isocratic elution program using a mobile phase consisting of HPLC-grade methanol: 5 mm ammonium acetate (80:20 v/v) at a flow-rate of 1.0 mL/min with a total runtime of 3.0 min. The proposed method has been validated with a linear range of 4-400 ng/mL for venlafaxine and 5-500 ng/mL for O-desmethyl venlafaxine. The method was applied for a bio-equivalence study of 75 mg tablets formulation in 32 Indian male healthy subjects under fasting conditions.

Comparison of postoperative intraocular pressure in patients with Densiron-68 vs conventional silicone oil: a case-control study.

UNLABELLED: A solution of perfluorohexyloctane and silicone oil with a specific gravity of 1.06 g/cm(3) (Densiron-68) has similar properties as conventional silicone oil (SO) in terms of the shape of the bubble and its ability to act as an internal tamponade agent. We conducted a case-control study to compare the postoperative intraocular pressure (IOP) in patients treated with Densiron-68 with those treated with SO. METHODS: Seventy-one eyes of 71 patients and 57 eyes of 57 patients who had received Densiron-68 and SO, respectively, were included in our study. Both groups were found to have matched for their preoperative comorbidities (diabetes, glaucoma, phakic status, and refractive errors). IOP at first day, between seventh and fourteenth day, and at 4 week postoperatively was recorded. RESULTS: The mean IOP was higher in patients treated with Densiron-68 at day 1 and between seventh and fourteenth day postoperatively (P=0.05 and 0.01, respectively). By the 4th week, the IOP difference between the two groups was insignificant (P=0.17). The difference in the two groups could still be clinically significant and the raised IOP in Densiron-68 group was more difficult to treat in some cases.On day 1, nine eyes (12.7%) in the Densiron-68 group and two eyes (3.5%) in the SO group had IOP greater than 30 mmHg. At 4 weeks, IOP of more than 30 mmHg was seen in nine eyes (12.7%) in the Densiron-68-treated group and in one eye (1.8%) in the SO group. CONCLUSION: The use of Densiron-68 was associated with a higher IOP in the early postoperative period when compared with SO.

Radioprotective properties of Hippophae rhamnoides (sea buckthorn) extract in vitro.

BACKGROUND: Hippophae rhamnoides, a high altitude habitat plant, has been extremely used in traditional medicinal practices for treating a variety of ailments. Recently, an extract (RH-3) prepared from berries of Hippophae rhamnoides has been reported to exhibit significant radioprotection against whole body lethal irradiation. OBJECTIVE: Present study was undertaken to elucidate the DNA binding ability of an extract (RH-3) prepared from berries of Hippophae rhamnoides and its role in modulating radiation induced frank and clustered DNA damage. METHOD: Agarose gel electrophoresis was employed as method to understand DNA binding potential and DNA protective ability of RH-3. RESULTS: RH-3 in a dose dependent fashion interacted with plasmid DNA (pUC18) reducing the mobility of supercoiled form and increasing the amount of the complex in the well indicating its ability to interact with plasmid DNA. RH-3 at higher concentrations (> 0.4 mg/ml) almost completely prevented the migration of supercoiled form without interfering with mobility of open circular form indicating its ability to selectively interact with supercoiled form. Studies done with supercoiled or open circular form also revealed the binding specificity of RH-3 for supercoiled form of plasmid. Both inhibited radiation induced strand breaks and DNA interaction by RH-3 were found to be dependent upon pH and the order of efficacy was found to be acidic pH> neutral pH > alkaline pH. RH-3 in a dose dependent manner inhibited radiation induced frank single, double strand breaks as well as endonuclease IV detectable abasic sites (clusters) and maximum reduction was observed at a concentration of 200 µg/ml. CONCLUSION: Results obtained in this study suggest that the ability of RH-3 to interact with DNA could be playing a significant role in preventing radiation induced DNA damage.

Neuropilins in physiological and pathological angiogenesis.

Neuropilin-1 (Np1) and neuropilin-2 (Np2) are transmembrane glycoproteins with large extracellular domains that interact with both class 3 semaphorins and vascular endothelial growth factor (VEGF), and are involved in the regulation of many physiological pathways, including angiogenesis. The neuropilins also interact directly with the classical receptors for VEGF, VEGF-R1 and -R2, mediating signal transduction. The heart, glomeruli and osteoblasts express both Np1 and Np2, but there is differential expression in the adult vasculature, with Np1 expressed mainly by arterial endothelium, whereas Np2 is only expressed by venous and lymphatic endothelium. Both neuropilins are commonly over-expressed in regions of physiological (wound-healing) and pathological (tumour) angiogenesis, but the signal transduction pathways, neuropilin-mediated gene expression and the definitive role of neuropilins in angiogenic processes are not fully characterized. This review details the current evidence for the role of neuropilins in angiogenesis, and suggests future research directions that may enhance our understanding of the mechanisms of action of this unique family of proteins.