Expression of 2-deoxy-<Emphasis Type="Italic">scyllo</Emphasis>-inosose synthase (<Emphasis Type="Italic">kan</Emphasis>A) from kanamycin gene cluster in <Emphasis Type="Italic">Streptomyces lividans</Emphasis>

An actinomycetes expression vector (pIBR25) was constructed and applied to express a gene from the kanamycin biosynthetic gene cluster encoding 2-deoxy-scyllo-inosose synthase (kanA) in Streptomyces lividans TK24. The expression of kanA in pIBR25 transformants reached a maximum after 72 h of culture. The plasmid pIBR25 showed better expression than pSET152, and resulted in the formation of insoluble KanA when it was expressed in Escherichia coli. This strategy thus provides a valuable tool for expressing aminoglycoside-aminocyclitols (AmAcs) biosynthetic genes in Streptomyces spp.     

Urea‐induced binding between diclofenac sodium and bovine serum albumin: a spectroscopic insight

We investigated the interaction of diclofenac sodium (Dic.Na) with bovine serum albumin (BSA) in the absence and presence of urea using different spectroscopic techniques. A fluorescence quenching study revealed that the Stern–Volmer quenching constant decreases in the presence of urea, decreasing further at higher urea concentrations. The binding constant and number of binding sites were also evaluated for the BSA–Dic.Na interaction system in the absence and presence of urea using a modified Stern–Volmer equation. The binding constant is greater at high urea concentrations, as shown by the fluorescence results. In addition, for the BSA–Dic.Na interaction system, a static quenching mechanism was observed, which was further confirmed using time‐resolved fluorescence spectroscopy. UV–vis spectroscopy provided information about the formation of a complex between BSA and Dic.Na. Circular dichroism was carried out to explain the conformational changes in BSA induced by Dic.Na in the absence and presence of urea. The presence of urea reduced the α‐helical content of BSA as the Dic.Na concentration varied. The distance r between the donor (BSA) and acceptor (Dic.Na) was also obtained in the absence and presence of urea, using fluorescence resonance energy transfer. Copyright © 2015 John Wiley & Sons, Ltd.     

Identification of novel genetic markers associated with clinical phenotypes of systemic sclerosis through a genome-wide association strategy

The aim of this study was to determine, through a genome-wide association study (GWAS), the genetic components contributing to different clinical sub-phenotypes of systemic sclerosis (SSc). We considered limited (lcSSc) and diffuse (dcSSc) cutaneous involvement, and the relationships with presence of the SSc-specific auto-antibodies, anti-centromere (ACA), and anti-topoisomerase I (ATA). Four GWAS cohorts, comprising 2,296 SSc patients and 5,171 healthy controls, were meta-analyzed looking for associations in the selected subgroups. Eighteen polymorphisms were further tested in nine independent cohorts comprising an additional 3,175 SSc patients and 4,971 controls. Conditional analysis for associated SNPs in the HLA region was performed to explore their independent association in antibody subgroups. Overall analysis showed that non-HLA polymorphism rs11642873 in IRF8 gene to be associated at GWAS level with lcSSc (P = 2.32×10⁻¹², OR = 0.75). Also, rs12540874 in GRB10 gene (P = 1.27 × 10⁻⁶, OR = 1.15) and rs11047102 in SOX5 gene (P = 1.39×10⁻⁷, OR = 1.36) showed a suggestive association with lcSSc and ACA subgroups respectively. In the HLA region, we observed highly associated allelic combinations in the HLA-DQB1 locus with ACA (P = 1.79×10⁻⁶¹, OR = 2.48), in the HLA-DPA1/B1 loci with ATA (P = 4.57×10⁻⁷⁶, OR = 8.84), and in NOTCH4 with ACA P = 8.84×10⁻²¹, OR = 0.55) and ATA (P = 1.14×10⁻⁸, OR = 0.54). We have identified three new non-HLA genes (IRF8, GRB10, and SOX5) associated with SSc clinical and auto-antibody subgroups. Within the HLA region, HLA-DQB1, HLA-DPA1/B1, and NOTCH4 associations with SSc are likely confined to specific auto-antibodies. These data emphasize the differential genetic components of subphenotypes of SSc.     

Development of an antibody against a 170-kDa fragment of fibroin isolated from cocoon fibres of Bombyx mori.

A simple method for the isolation of denatured fragments of the fibroin protein from cocoon fibres by alkali solubilization is discussed. This 170-kDa antigen has been purified and used to raise the polyclonal antibody in rabbit. The specificity of this antibody to the purified cocoon protein has suggested strong immunoreactivity up to a titre of 1:5000 dilution of the antibody. Further, dot-blot analysis with the tissue extracts from silk glands of different larval stages (3rd to 5th) reveals that this antibody reacts showing a stage-specific increase in the intensity of the colour, correlating well with the in-vivo expression of the silk protein. This study suggests the availability of a specific polyclonal antibody that detects the native fibroin with no crossreactivity with other tissue proteins.     

Expression and purification of mouse TIMP-1 from E. coli.

Tissue inhibitors of metalloproteinases (TIMPs) constitute a family of secreted glycoproteins involved in regulating extracellular matrix degradation in both normal and malignant tissues. We have expressed a cDNA clone of mouse TIMP-1 as a 22-kDa protein with 12 cysteine residues in E. coli and purified protein that shows inhibitory activity against collagenase following renaturation by chemical means. The low specific activity and circular dichroism measurements suggest, however, that the renaturation of the mouse recombinant (non-glycosylated) protein is not efficient under the conditions we have used, indicative of either thermodynamic instability or the transition to kinetic intermediates which have very low in vitro refolding rates.     

Lead-210 content of food samples in India

Summary The paper presents results of measurements made on cereals and composite meal samples collected from Bombay market for their lead-210 content. The details of sampling and analytical chemistry procedures are also given. The Pb-210 contents of most of these samples were in the range of 1–5 pCi/kg of cereals samples. The concentrations in composite meal samples were mostly in the range of 1–3.5 pCi of²¹⁰Pb per composite meal. The assessment of daily intake of this isotope through food-stuffs has been made.     

Integrated Nutrient Management Improves Productivity and Quality of Sugarcane (<i>Saccharum Officinarum</i> L.)

Sugarcane is one of the major important sugar yielding crops in Bangladesh. As an exhaustive crop, sugarcane removes a huge amount of plant nutrients from the soil. However, the combined use of organic and inorganic fertilizers can be a good approach to deal with nutrient depletion and promote sustainable crop production as well as improve soil health. Therefore, an attempt was made to identify the most fruitful and profitable integrated nutrient management on the aspects of growth, yield and quality of sugarcane in two consecutive growing seasons. Seven treatments: T₁=Control, T₂=165:55:120:30:10:2.5:4 kg N:P:K:S:Mg:Zn:B ha⁻¹, T₃=Poultry Litter (PL) at 5 t ha⁻¹+95:51:87:9:10:2.5:4 kg N:P:K:S:Mg:Zn:B ha⁻¹, T₄=Cow Dung (CD) at 15 t ha⁻¹+ 36:52:60:17:10:2.5:4 kg N:P:K:S:Mg:Zn:B ha⁻¹, T₅=Press Mud (PM) at 15 t ha⁻¹+10:50:43:0:10:2.5:4 kg N:P:K:S:Mg:Zn:B ha⁻¹, T₆=Mustard Oil Cake (MOC) at 0.5 t ha⁻¹+140:54:115:25:10:2.5:4 kg N:P:K:S:Mg:Zn:B ha⁻¹ and T₇=GM (Green Manure) at 5 t ha⁻¹+140:53:100:28:10:2.5:4 kg N:P:K:S:Mg:Zn:B ha⁻¹ were used in this experiment. Two years data showed that treatment T₃ produced the maximum amount of tillers, total dry matter yield, millable sugarcane, cane yield and sugar yield, followed by the T₄ treatment. The highest stalk heights were recorded in the T₃ treatment, which was statistically similar to all other treatments except T₁ and T₂. The juice quality parameters viz., brix and pol in cane were found significant in treatment T₃ while the highest purity was obtained in the T₇ treatment. All the data of Jaggery (goor) quality parameters, the highest sucrose content, color transmittance, Jaggery (goor) recovery and the lowest ash content of Jaggery (goor) were observed in the T₃ treatment, which was statistically similar to the T₄ treatment in both seasons. The highest cost of production was obtained from the T₆ treatment while the highest gross return, net return and BCR were recorded in the T₃ treatment. No significant changes were found in one cycle of sugarcane in initial and post-harvest soil characteristics viz., pH, organic carbon, total N, and available P, K and S contents due to integrated use of different fertilizer packages. From the experimental findings, it was concluded that treatment T₃ followed by T₄ treatment would be the better productive and profitable integrated nutrient management technology for ensuring higher yields and quality of sugarcane without soil fertility degradation in the High Ganges River Floodplain soils.     

Improvements to the Red List Index

The Red List Index uses information from the IUCN Red List to track trends in the projected overall extinction risk of sets of species. It has been widely recognised as an important component of the suite of indicators needed to measure progress towards the international target of significantly reducing the rate of biodiversity loss by 2010. However, further application of the RLI (to non-avian taxa in particular) has revealed some shortcomings in the original formula and approach: It performs inappropriately when a value of zero is reached; RLI values are affected by the frequency of assessments; and newly evaluated species may introduce bias. Here we propose a revision to the formula, and recommend how it should be applied in order to overcome these shortcomings. Two additional advantages of the revisions are that assessment errors are not propagated through time, and the overall level extinction risk can be determined as well as trends in this over time.