A Novel Enediynyl Peptide Inhibitor of Furin That Blocks Processing of proPDGF-A, B and proVEGF-C

BACKGROUND: Furin represents a crucial member of secretory mammalian subtilase, the Proprotein Convertase (PC) or Proprotein Convertase Subtilisin/Kexin (PCSK) superfamily. It has been linked to cancer, tumorgenesis, viral and bacterial pathogenesis. As a result it is considered a major target for intervention of these diseases. METHODOLOGY/PRINCIPAL FINDINGS: Herein, we report, for the first time, the synthesis and biological evaluation of a newly designed potent furin inhibitor that contains a highly reactive beta-turn inducing and radical generating "enediynyl amino acid" (Eda) moiety. "Eda" was inserted between P1 and P1' residues of hfurin(98-112) peptide, derived from the primary cleavage site of furin's own prodomain. The resulting hexadecapeptide derivative inhibited furin in vitro with IC(50) approximately 40 nM when measured against the fluorogenic substrate Boc-RVRR-MCA. It also inhibited furin-mediated cleavage of a fluorogenic peptide derived from hSARS-CoV spike protein with IC(50) approximately 193 nM. Additionally it also blocked furin-processing of growth factors proPDGF-A, B and VEGF-C that are linked to tumor genesis and cancer. Circular dichroism study showed that this inhibitor displayed a predominantly beta-turn structure while western blots confirmed its ability to protect furin protein from self degradation. CONCLUSION/SIGNIFICANCE: These findings imply its potential as a therapeutic agent for intervention of cancer and other furin-associated diseases

Functions of interleukin-8 are mediated through thiol group(s) of IL-8 receptor in human polymorphonuclear neutrophils Effects of 5,5’-dithio-bis(2-nitrobenzoic acid) on IL-8 receptor

Interleukm-8, a neutrophil chemotacttc agent causes excesstve accumulation of the cells in a number of inflammatory diseases. The activity has been shown to be mediated through a specific functional receptor present on the surface of neutrophils. No information is available about the ammo acids constitutmg the IL-8 binding domain of the receptor. Treatment of neutrophtls with 5.5’-dtthio-bts(3-mtrobenzotc acid), a thiol-specific modifier. at the concentrattons of 0.4 mM and 1 mM reduced IL-8 binding ability and IL-g-induced migration of the cells by 45% and 65%.respectively. Dithiothrettol could regenerate the bmding capacity and the ligand could protect the receptor from the effect of the reagent. All the evidence suggests that one or more critical thiol residues are located m the IL-8 bmdmg site of the receptor which are mdtspensible for normal functions of IL-8

Status epilepticus as the only presentation of the neonatal Bartter syndrome

Bartter syndrome is a rare hereditary (autosomal recessive) salt-losing tubulopathy characterized by hypokalemia, hypochloremia, metabolic alkalosis, and normal blood pressure with hyperreninemia, The underlying renal abnormality results in excessive urinary losses of sodium, chloride, and potassium. We report a case of a four-month-old infant with neonatal Bartter syndrome, who presented only with status epilepticus. To the best of our present knowledge, there is no reported case of Bartter syndrome who presented with status epilepticus

New automated fluoroscopic systems for pediatric applications.” J Appl Clin Med Phys 6:88–105

Pediatric patients are at higher risk to the adverse effects from exposure to ionizing radiation than adults. The smaller sizes of the anatomy and the reduced X-ray attenuation of the tissues provide special challenges. The goal of this effort is to investigate strategies for pediatric fluoroscopy in order to minimize the radiation exposure to these individuals, while maintaining effective diagnostic image quality. Modern fluoroscopy systems are often entirely automated and computer controlled. In this paper, various selectable and automated modes are examined to determine the influence of the fluoroscopy parameters upon the patient radiation exposures and image quality. These parameters include variable X-ray beam filters, automatic brightness control programs, starting kilovolt peak levels, fluoroscopic pulse rates, and other factors. Typical values of radiation exposure rates have been measured for a range of phantom thicknesses from 5 cm to 20 cm of acrylic. Other factors that have been assessed include spatial resolution, lowcontrast discrimination, and temporal resolution. The selection menu for various procedures is based upon the examination type, anatomical region, and patient size. For pediatric patients, the automated system can employ additional filtration, special automatic brightness control curves, pulsed fluoroscopy, and other features to reduce the patient radiation exposures without significantly compromising the image quality. The benefits gained from an optimal selection of automated programs and settings for fluoroscopy include ease of operation, better image quality, and lower patient radiation exposures. PACS numbers: 87.59.-e, 87.62.+n Key words: digital fluoroscopy, pediatric radiation exposur

Biostimulation of indigenous microbial community for bioremediation of petroleum refinery sludge.

Nutrient deficiency severely impairs the catabolic activity of indigenous microorganisms in hydrocarbon rich environments (HREs) and limits the rate of intrinsic bioremediation. The present study aimed to characterize the microbial community in refinery waste and evaluate the scope for biostimulation based in situ bioremediation. Samples recovered from the wastewater lagoon of Guwahati refinery revealed a hydrocarbon enriched high total petroleum hydrocarbon (TPH), oxygen-, moisture-limited, reducing environment. Intrinsic biodegradation ability of the indigenous microorganisms was enhanced significantly (>80% reduction in TPH by 90 days) with nitrate amendment. Preferred utilization of both higher- (>C30) and middle- chain (C20-30) length hydrocarbons were evident from GC-MS analysis. Denaturing gradient gel electrophoresis (DGGE) and community level physiological profiling (CLPP) analyses indicated distinct shift in community’s composition and metabolic abilities following nitrogen (N) amendment. High throughput deep sequencing of 16S rRNA gene showed that the native community was mainly composed of hydrocarbon degrading, syntrophic, methanogenic, nitrate/iron/sulfur reducing facultative anaerobic bacteria and archaebacteria, affiliated to γ- and δ-Proteobacteria and Euryarchaeota respectively. Genes for aerobic and anaerobic alkane metabolism (alkB and bssA), methanogenesis (mcrA), denitrification (nirS and narG) and N2 fixation (nifH) were detected. Concomitant to hydrocarbon degradation, lowering of dissolve O2 and increase in oxidation-reduction potential (ORP) marked with an enrichment of N2 fixing, nitrate reducing aerobic/facultative anaerobic members e.g., Azovibrio, Pseudoxanthomonas and Commamonadaceae members was evident in N amended microcosm. This study highlighted that indigenous community of refinery sludge was intrinsically diverse, yet appreciable rate of in situ bioremediation could be achieved by supplying adequate N sources

Petroleum hydrocarbon rich oil refinery sludge of North-East India harbours anaerobic, fermentative, sulfate-reducing, syntrophic and methanogenic microbial populations

Abstract Background Sustainable management of voluminous and hazardous oily sludge produced by petroleum refineries remains a challenging problem worldwide. Characterization of microbial communities of petroleum contaminated sites has been considered as the essential prerequisite for implementation of suitable bioremediation strategies. Three petroleum refinery sludge samples from North Eastern India were analyzed using next-generation sequencing technology to explore the diversity and functional potential of inhabitant microorganisms and scope for their on-site bioremediation. Results All sludge samples were hydrocarbon rich, anaerobic and reduced with sulfate as major anion and several heavy metals. High throughput sequencing of V3-16S rRNA genes from sludge metagenomes revealed dominance of strictly anaerobic, fermentative, thermophilic, sulfate-reducing bacteria affiliated to Coprothermobacter, Fervidobacterium, Treponema, Syntrophus, Thermodesulfovibrio, Anaerolinea, Syntrophobacter, Anaerostipes, Anaerobaculum, etc., which have been well known for hydrocarbon degradation. Relatively higher proportions of archaea were detected by qPCR. Archaeal 16S rRNA gene sequences showed presence of methanogenic Methanobacterium, Methanosaeta, Thermoplasmatales, etc. Detection of known hydrocarbon utilizing aerobic/facultative anaerobic (Mycobacterium, Pseudomonas, Longilinea, Geobacter, etc.), nitrate reducing (Gordonia, Novosphigobium, etc.) and nitrogen fixing (Azovibrio, Rhodobacter, etc.) bacteria suggested niche specific guilds with aerobic, facultative anaerobic and strict anaerobic populations. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) predicted putative genetic repertoire of sludge microbiomes and their potential for hydrocarbon degradation; lipid-, nitrogen-, sulfur- and methane- metabolism. Methyl coenzyme M reductase A (mcrA) and dissimilatory sulfite reductase beta-subunit (dsrB) genes phylogeny confirmed methanogenic and sulfate-reducing activities within sludge environment endowed by hydrogenotrophic methanogens and sulfate-reducing Deltaproteobacteria and Firmicutes members. Conclusion Refinery sludge microbiomes were comprised of hydrocarbon degrading, fermentative, sulfate-reducing, syntrophic, nitrogen fixing and methanogenic microorganisms, which were in accordance with the prevailing physicochemical nature of the samples. Analysis of functional biomarker genes ascertained the activities of methanogenic and sulfate-reducing organisms within sludge environment. Overall data provided better insights on microbial diversity and activity in oil contaminated environment, which could be exploited suitably for in situ bioremediation of refinery sludge

Layering smart management practices to sustainably maintain rice yields and improve water use efficiency in eastern India

Rice in eastern India is widely cultivated using conventional management practices, including puddled transplanting (PTR) for crop establishment, which are water and energy intensive. To conserve water and maximise its efficient use, improved crop management practices must be identified which are feasible and attractive to farmers. These improved practices include dry direct seeding of rice (dDSR), improved medium-duration rice varieties and alternate wetting and drying (AWD) irrigation management. These have separately been shown to reduce the irrigation water requirement for rice cultivation while maintaining yields similar to those of traditional high yielding long-duration varieties. We conducted a field experiment over two rice-growing seasons to examine the effects on rice grain yield and water productivity of layering these improved management practices, compared to conventional PTR with traditional long-duration rice. We also assessed the usefulness of simple perforated tubes to implement AWD management in the field. dDSR reduced rice grain yield by 3–6% while requiring 23% less irrigation water over PTR. The medium-duration hybrid rice variety produced 2–4% higher grain yield under direct seeding while using 4–5% less irrigation water than the long-duration variety. AWD reduced irrigation water use by 26–52% but also reduced grain yield by 3–14%. Water use efficiency was improved by dDSR, AWD and by using medium-duration hybrid variety. Combining all three management practices reduced irrigation water use by an average of 18% while improving irrigation water productivity by 21%, without significant reduction in grain yield under shallow AWD of 10 cm depth in dDSR and upto 15 cm depth for PTR. This combination is a novel approach to sustainably improve farmers’ rice productivity. Additionally, simple perforated tubes are an effective and innovative tool for farmers to implement AWD management. Our research suggests that farmers in eastern India and similar rice-growing agro-ecologies could reduce the irrigation water applied and increase water use efficiency while maintaining rice crop productivity by combining dry direct seeding with a medium-duration rice variety under AWD irrigation management