Design of joint differential for transformation of two rotary movements to linear motion.

Tato práce si klade za cíl navrhnout mechanismus diferenciálního propojení, který poskytuje řadu variací zdvihu. Je proveden výzkum a jsou navrženy různé varianty. 3D model diferenciálního mechanismu zabudovaného v PTC kreo parametrickém systému. Jedná se o sedmibodový mechanismus, který ke svému provozu vyžaduje dva pohony. Byla provedena kinematická analýza pro určení dimenze vazeb v modelu a požadovaný výstup, takže z jednoho mechanismu jsou možné různé hodnoty variace zdvihu. Dynamická analýza prováděná pomocí PTC parametrického mechanismu pro analýzu sil v jejich spojích. Analýza napětí pomocí PTC kreo parametrické simulace pro sledování chování mechanismu napětí a posunutí. CAD dokumentace finálního modelu hotová a zkontrolovaná.This thesis aims to design a differential linkage mechanism that provides a range of stroke variation. Research done and different variants are proposed. The 3D model of differential mechanism built in PTC creo parametric system. It is a seven bar mechanism which requires two drives for its operation. Kinematic analysis done for determining the dimension of the links in the model and the required output achieved so that different values of stroke variation is possible from single mechanism. Dynamic analysis done using PTC creo parametric mechanism for analysing forces at their joints. Stress analysis done using PTC creo parametric simulation for observing the stress and displacement behaviour of the mechanism. CAD documentation of final model done and reviewed

Human annexin A6 interacts with influenza a virus protein M2 and negatively modulates infection

Copyright © 2012, American Society for Microbiology. All Rights ReservedThe influenza A virus M2 ion channel protein has the longest cytoplasmic tail (CT) among the three viral envelope proteins and is well conserved between different viral strains. It is accessible to the host cellular machinery after fusion with the endosomal membrane and during the trafficking, assembly, and budding processes. We hypothesized that identification of host cellular interactants of M2 CT could help us to better understand the molecular mechanisms regulating the M2-dependent stages of the virus life cycle. Using yeast two-hybrid screening with M2 CT as bait, a novel interaction with the human annexin A6 (AnxA6) protein was identified, and their physical interaction was confirmed by coimmunoprecipitation assay and a colocalization study of virus-infected human cells. We found that small interfering RNA (siRNA)-mediated knockdown of AnxA6 expression significantly increased virus production, while its overexpression could reduce the titer of virus progeny, suggesting a negative regulatory role for AnxA6 during influenza A virus infection. Further characterization revealed that AnxA6 depletion or overexpression had no effect on the early stages of the virus life cycle or on viral RNA replication but impaired the release of progeny virus, as suggested by delayed or defective budding events observed at the plasma membrane of virus-infected cells by transmission electron microscopy. Collectively, this work identifies AnxA6 as a novel cellular regulator that targets and impairs the virus budding and release stages of the influenza A virus life cycle.This work was supported by the Research Fund for the Control of Infectious Disease (project 09080892) of the Hong Kong Government, the Area of Excellence Scheme of the University Grants Committee (grant AoE/M-12/-06 of the Hong Kong Special Administrative Region, China), the French Ministry of Health, the RESPARI Pasteur Network

Cofilin-1 is involved in regulation of actin reorganization during influenza A virus assembly and budding

Influenza A virus (IAV) assembly and budding on host cell surface plasma membrane requires actin cytoskeleton reorganization. The underlying molecular mechanism involving actin reorganization remains unclarified. In this study, we found that the natural antiviral compound petagalloyl glucose (PGG) inhibits F-actin reorganization in the host cell membrane during the late stage of IAV infection, which are associated with the suppression of total cofilin-1 level and its phosphorylation. Knock-down of cofilin-1 reduces viral yields. These findings provide the first evidence that cofilin-1 plays an important role in regulating actin reorganization during IAV assembly and budding

The Effect of Cortical Spreading Depression Events on Anti-Migraine Agent Uptake Via Regulation of Functional Expression of OATP1A2/OATP1A4 at the Blood Brain Barrier

The blood brain barrier (BBB), regulated by the neurovascular unit (NVU), restricts movement of substances to and from the blood to the brain, posing a daunting challenge for drug design where effective uptake is critical to a drug’s efficacy and safety. Previous research has shown that cortical spreading depression (CSD) events may be associated with migraine and other neurological disorders. What has not been well studied is how CSD pathological conditions, including associated changes in potassium ion levels and the release of mediators such as calcitonin gene related peptide (CGRP), may affect BBB integrity at the level of the microvessels, and consequently, uptake of brain-targeting therapeutics such as Triptans and Gepants. Xenobiotic transport occurs by paracellular, transcellular and/or transport-mediated processes, including by the solute carrier organic anion transporter, OATP. OATP1A2 (human)/ Oatp1a4 (rodent) are widely expressed bidirectional sodium-independent solute carriers involved in transport of a wide range of amphipathic substrates. Previous studies have shown OATP expression and activity may be pathologically regulated during inflammatory and/or hypoxic/reoxygenation states. Previously, we showed CSD promotes paracellular leak as determined by sucrose uptake as well as transporter facilitated uptake of Sumatriptan. Herein, we first investigated how high abluminal potassium levels, in vitro, and potassium chloride (KCl)-induced CSD events, in vivo, impact Oatp1a4 functional expression and uptake of Sumatriptan following prophylactic or abortive administration. Oatp1a4 facilitated parenchymal uptake was observed when Sumatriptan was dosed prior, but not after, CSD induction suggesting subclinical insults can rapidly change Oatp1a4 functional expression and its role in Sumatriptan uptake relies on timing of each dosing paradigm. Next, to investigate how CSD events promote paracellular leak, tight junction reorganization and regulation of transporter functional expression, we tested whether CSD elevates local CGRP leading to uptake of Rimegepant via paracellular and Oatp1a4 transport. Several novel findings were identified including: (1) CSD induction with a single cortical injection of KCl did not significantly change CGRP levels in the V1M portion of the occipital cortex (V1M cortex), plasma, Trigeminal nucleus caudalis (Vc) or Periaqueductal grey (PAG) of mice; (2) brief and acute increases in CGRP induced significant decreases in Transendothelial electrical resistance (TEER), but not a functional breach of BEB paracellular integrity; (3) Sustained Rimegepant produced delayed paracellular leak absent in the presence of CGRP; (4) CGRP dose-dependently altered Oatp1a4 expression in bEnd.3 cells; and (5) sucrose uptake increased in KCl- pulsed cells after sustained exposure to Rimegepant, but Rimegepant is not detected after during KCl pulse, suggesting that the size of paracellular leak during CSD as previously reported is < 1200g/mol (MW Rimegepant succinate),, narrowing the paracellular leak size during CSD from 4000g/mol. Finally, we hypothesized that the CGRP receptor could be selectively targeted at the CLR-RAMP1 interaction to reduce side effects and drug-drug interactions. We used in silico molecular modeling techniques and identified potential novel CGRP receptor targeting compounds. Quantitative functional analysis was performed by measuring nitrate levels in bEND.3 cells treated with the top four compounds; however data was inconclusive requiring future studies with other methods to validate these agents.Release after 08/29/202

Anaerobic Process for Biohydrogen Production using Keratin Degraded Effluent

Fossil fuel energy depletion and removal of environmental wastes are big problem in many countries. Some of the wastes contain a considerable amount of protein and carbon compounds; little concentration is given to utilizing or recycling these wastes in a technological way for production of renewable energy. Feathers are generated in large amounts as waste at commercial poultry processing plant, reaching millions of tons per year worldwide. Feathers are rich in keratin protein. In this work, keratin was degraded and the degraded effluent was evaluated for biohydrogen production by anaerobic (dark fermentation) process. The keratin degrading microorganisms were isolated from the soil sample where the feather wastes were disposed using minimal media with prepared keratin powder. The isolates were named as KHAD1, KHAD2 and KHAD3. The isolates were identified as Serratia marcescens KHAD1 (MH422128), Bacillus subtilis KHAD2 (MH422129) and Pseudomonas aeruginosa KHAD3 (MH422009) based on their 16S rRNA sequencing. The isolates were screened for biodegradability of keratin protein and higher biodegrading potential microorganism such as Pseudomonas aeruginosa KHAD3. The optimization studies for biohydrogen production using isolates in keratin degraded effluent at various pH (1, 2, 4, 6, 8, 10 & 11). The maximum biohydrogen production was obtained at pH 11 (210 ml/L) by Pseudomonas aeruginosa KHAD3

A simple fluorescent sensor for the meticulous recognition of Cu2+ ion and its functioning in logic gate and keypad lock

A highly selective and perceptive “off–on” mode fluorescent receptor (E)-1-(5-methylfuran-2-yl)-N-(2-(phenylthio)phenyl)methanimine (R) has been constructed for the expeditious recognition of Cu2+ ions in the semi-aqueous medium. The strong fluorescence was observed at 468 nm for Cu2+ ions over other tested metal ions by R, which could be attributed to the constraint of the CHdouble bondN isomerization mechanism and provoked chelation enhanced fluorescence effect (CHEF) upon interaction with copper ions. According to Job’s plot, the binding proportion between R and copper ion is 2:1 and further validated by mass analysis and quantum chemical studies. Based on the emission titration, the limits of detection (LOD) were determined to be 0.194 μM. The obtained results designate that the receptor R serves as a significant nominee for an expeditious recognition of Cu2+ ions. Further, R was fruitfully useful in molecular keypad locks, and molecular logic gate circuits and it could be applied to find out the trace quantity of Cu2+ ions in water samples

Inactivation of the p53 Gene by Either Mutation or HPV Infection is Extremely Frequent in Human Oral Squamous Cell Carcinoma Cell Lines

The state of p53 tumour suppressor and the frequency of high-risk human papillomavirus (HPV) infections were studied in nine human oral cancer cell lines. Three cancer cell lines (SCC-4, Tu-177 and FaDu) had similar amounts of p53 transcripts to normal cells, but contained significantly higher levels of p53 protein than the normal control cells. Sequencing highly conserved open reading frames of the p53 gene of these cancer cells showed point mutations in the SCC-4 and Tu-177 cell lines, a base transition from CCC to TCC occurred at codon 151; and in the line FaDu, a mutation of CGG to CTG occurred at codon 248. The HEp-2 and 1483 cancer lines contained significantly lower levels of p53 protein compared to the normal counterpart. Sequencing of p53 cDNA for HEp-2 and 1483 lines showed no mutations, but northern analysis revealed that these cell lines expressed HPV-18 E6/E7 messages. Four cell lines (SCC-9, SCC-15, SCC-25, and Tu-139) expressed negligible amounts of p53 transcripts compared to the normal counterpart and undetectable levels of p53 protein. These cell lines contained mutations in the highly conserved open reading frames of the p53 gene as follows: the SCC-9 had a deletion of 32 base pairs between codons 274 and 285; the line SCC-15 had an insertion of five base pairs between codons 224 and 225; the line SCC-25 had a deletion of two base pairs in codon 209; and the Tu-139 line had a deletion of 46 base pairs between codons 171 and 186. These data indicate that aberrant expression of p53 and altered levels of p53 protein are extremely frequent in oral cancer cell lines, and that these changes may result from either mutations or infection with high-risk HPV

Acinetobacter junii AH4-A Potential Strain for Biohydrogen Production from Dairy Industry Anaerobic Sludge

The present study aims to enhance the efficiency of anaerobic sludge microorganisms to produce hydrogen (H2 ) through various pre-treatment methods. The various pre-treatment methods such as base, acid, chloroform, heat shock, freezing and thawing have enabled to isolate acidogenic bacteria with higher bio H2 producing activity in an anoxic environment. From these various treatments, bacteria were isolated and screened for bio H2 capabilities. Among the one bacterial strain, AH4 strain showed maximum cumulative H2 production and Hydrogen Yield (HY) using 100% diary anaerobic sludge. AH4 strain was identified as Acinetobacter junii using 16S rRNA gene sequence and used for further experimental analysis. Biohydrogen productions of Acinetobacter junii were measured at different experimental setup such as various pH levels (5.0, 5.5, 6.0, 6.5, 7.0, 7.5 and 8.0) and different substrate concentration (10 - 100%) of dairy anaerobic sludge substrate. At pH 7.5 and 60% substrate concentration, the strain AH4 Acinetobacter junii displayed the maximum cumulative H2 production of 945.7 ml/L and H2 yield 1.35 mol H2 / mol glucose. Based on our results, we concluded that Acinetobacter junii can be used as a promising bio agent for hydrogen production on a large scale using dairy anaerobic sludge as substrate

Frequent Mutations of p53 and MTS1/CDK4I Tumor Suppressor Genes in Chinese Preneoplastic and Neoplastic Oral Tissues

This study is, in part, supprtred by grants from the Smokeless Tobacoo Research Council, Inc.(Number 0231) and USPHS research grants P50 DE100598 and RO1 DE10049 from the National Institute of Dental Research, NIH

Optimization (Substrate and pH) and Anaerobic Fermentative Hydrogen Production by Various Industrial Wastes Isolates Utilizing Biscuit Industry Waste as Substrate

The bio hydrogen (H2 ) production by anaerobic digestion of industrial waste is beneficial one due to the availability of proteins and carbohydrates as potential substrate for biological H2 production. An anaerobic fermentative route is a promising method of biohydrogenation. The microbial isolates from various industrial wastes (dairy, sugar and food) were assessed for their potential bio H2 production. The selected individual isolates (F1 - Bacillus subtilis and A3 -Bacillus subtilis) and their cocultures were used for the optimization of bio H2 production utilizing various concentration of biscuit industry waste as substrate at various pH conditions. The mixed consortium which displayed the higher bio H2 production was selected for the detailed analysis of the 3L fermentation studies using 90% Organic Loading Rate (OLR) substrate at pH 6.5. Significantly higher Hydrogen Yield (HY) of 0.87 mol H2 /mol glucose on 16th day of incubation was observed