Detection of Wolbachia endobacteria in Culex quinquefasciatus by Gimenez staining and confirmation by PCR

Background & objectives: Wolbachia are common intracellular bacteria that are found in arthropods and nematodes.These endosymbionts are transmitted vertically through host eggs and alter host biology in diverse ways, includingthe induction of reproductive manipulations, such as feminization, parthenogenesis, male killing and sperm-eggincompatibility. Since they can also move horizontally across species boundaries, Wolbachia is gaining importancein recent days as it could be used as a biological control agent to control vector mosquitoes or for paratransgenicapproaches. However, the study of Wolbachia requires sophisticated techniques such as PCR and cell culturefacilities which cannot be affordable for many laboratories where the diseases transmitted by arthropod vectorsare common. Hence, it would be beneficial to develop a simple method to detect the presence of Wolbachia inarthropods.Method: In this study, we described a method of staining Wolbachia endobacteria, present in the reproductivetissues of mosquitoes. The reliability of this method was compared with Gram staining and PCR based detection.Results: The microscopic observation of the Gimenez stained smear prepared from the teased ovary of wildcaught and Wolbachia (+) Cx. quinquefasciatus revealed the presence of pink coloured pleomorphic cells ofWolbachia ranging from cocci, comma shaped cells to bacillus and chain forms. The ovaries of Wolbachia (–)cured mosquito did not show any cell. Although Gram’s staining is a reliable differential staining for the otherbacteria, the bacterial cells in the smears from the ovaries of wild caught mosquitoes did not take the stain properlyand the cells were not clearly visible. The PCR amplified product from the pooled remains of wild caught andWolbachia (+) Cx. quinquefasciatus showed clear banding, whereas, no banding was observed for the negativecontrol (distilled water) and Wolbachia (–) Cx. quinquefasciatus.Interpretation & conclusion: The Gimenez staining technique applied, could be used to detect the members of theendobacteria Wolbachia easily, even in a simple laboratory without any special facilities or even in the fieldcondition and for handling large number of samples in a shorter duratio

Signaling Mechanisms in the Regulation of Renal Matrix Metabolism in Diabetes

Renal hypertrophy and accumulation of extracellular matrix proteins are among cardinal manifestations of diabetic nephropathy. TGF beta system has been implicated in the pathogenesis of these manifestations. Among signaling pathways activated in the kidney in diabetes, mTOR- (mammalian target of rapamycin-)regulated pathways are pivotal in orchestrating high glucose-induced production of ECM proteins leading to functional and structural changes in the kidney culminating in adverse outcomes. Understanding signaling pathways that influence individual matrix protein expression could lead to the development of new interventional strategies. This paper will highlight some of the diverse components of the signaling network stimulated by hyperglycemia with an emphasis on extracellular matrix protein metabolism in the kidney in diabetes

National instruments labVIEW and video imaging technique for health status monitoring

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Finite element modelling of four edges simply supported steel plate under impact load

No Abstract

Load Balanced Clustering Technique in MANET using Genetic Algorithms

Mobile adhoc network (MANET) has characteristics of topology dynamics due to factors such as energy conservation and node movement that leads to dynamic load-balanced clustering problem (DLBCP). Load-balancing and reliable data transfer between all the nodes are essential to prolong the lifetime of the network. MANET can also be partitioned into clusters for maintaining the network structure. Generally, Clustering is used to reduce the size of topology and to accumulate the topology information. It is necessary to have an effective clustering algorithm for adapting the topology change. In this, we used energy metric in genetic algorithm (GA) to solve the DLBCP. It is important to select the energy- efficient cluster head for maintaining the cluster structure and balance the load effectively. In this work, we used genetic algorithms such as elitism based immigrants genetic algorithm (EIGA) and memory enhanced genetic algorithm (MEGA) to solve DLBCP. These schemes select an optimal cluster head by considering the distance and energy parameters. We used EIGA to maintain the diversity level of the population and MEGA to store the old environments into the memory. It promises the load -balancing in cluster structure to increase the lifetime of the network. Experimental results show that the proposed schemes increases the network lifetime and reduces the total energy consumption. The simulation results show that MEGA and EIGA give a better performance in terms of load-balancing

Proteomic profiling of Burkholderia cenocepacia clonal isolates with different virulence potential retrieved from a cystic fibrosis patient during chronic lung infection

Respiratory infections with Burkholderia cepacia complex (Bcc) bacteria in cystic fibrosis (CF) are associated with a worse prognosis and increased risk of death. In this work, we assessed the virulence potential of three B. cenocepacia clonal isolates obtained from a CF patient between the onset of infection (isolate IST439) and before death with cepacia syndrome 3.5 years later (isolate IST4113 followed by IST4134), based on their ability to invade epithelial cells and compromise epithelial monolayer integrity. The two clonal isolates retrieved during late-stage disease were significantly more virulent than IST439. Proteomic profiling by 2-D DIGE of the last isolate recovered before the patient's death, IST4134, and clonal isolate IST439, was performed and compared with a prior analysis of IST4113 vs. IST439. The cytoplasmic and membrane-associated enriched fractions were examined and 52 proteins were found to be similarly altered in the two last isolates compared with IST439. These proteins are involved in metabolic functions, nucleotide synthesis, translation and protein folding, cell envelope biogenesis and iron homeostasis. Results are suggestive of the important role played by metabolic reprogramming in the virulence potential and persistence of B. cenocepacia, in particular regarding bacterial adaptation to microaerophilic conditions. Also, the content of the virulence determinant AidA was higher in the last 2 isolates. Significant levels of siderophores were found to be secreted by the three clonal isolates in an iron-depleted environment, but the two late isolates were more tolerant to low iron concentrations than IST439, consistent with the relative abundance of proteins involved in iron uptake.This work was supported by FEDER and FCT – Fundação para a Ciência e a Tecnologia (contract PEst-OE/EQB/LA0023/2011_ research line: Systems and Synthetic Biology; PhD grant to A.M. – SFRH/BD/37012/2007, and PD grants to S.S. – SFRH/BPD/75483/2010 and C.C. – SFRH/BPD/ 81220/2011. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.info:eu-repo/semantics/publishedVersio

BAT3 Guides Misfolded Glycoproteins Out of the Endoplasmic Reticulum

Secretory and membrane proteins that fail to acquire their native conformation within the lumen of the Endoplasmic Reticulum (ER) are usually targeted for ubiquitin-dependent degradation by the proteasome. How partially folded polypeptides are kept from aggregation once ejected from the ER into the cytosol is not known. We show that BAT3, a cytosolic chaperone, is recruited to the site of dislocation through its interaction with Derlin2. Furthermore, we observe cytoplasmic BAT3 in a complex with a polypeptide that originates in the ER as a glycoprotein, an interaction that depends on the cytosolic disposition of both, visualized even in the absence of proteasomal inhibition. Cells depleted of BAT3 fail to degrade an established dislocation substrate. We thus implicate a cytosolic chaperone as an active participant in the dislocation of ER glycoproteins.United States. National Institutes of HealthBoehringer Ingelheim Fond

Improvement of Weld Bead Characteristics in Gas Metal Arc Welding of SA515 Carbon Steel by Applying Alternating Shielding Gas Flow Technique

High service temperature of pressure vessel components necessitates the use of welded SA515 grade carbon steel components. The gas metal arc welding (GMAW) process using CO2 as shielding gas is known for its undesirable spatter behaviour and inferior weld quality. The alternating shielding gas flow (ASGF) technique is proposed in this study using the shielding gases, viz. CO2 and argon to overcome this difficulty. The welding current, stand-off distance, and shielding gas flow were all varied to improve the bead-on-plate profile geometry. The bead profile parameters such as depth of penetration, bead width, and bead height are considered as weld bead parameters. The following methods are used: correlation analysis, analysis of variance (ANOVA), modelling, and grey relational analysis (GRA). According to the findings, the welding current and ASGF are the most influential parameters impacting the weld bead characteristics. By increasing the welding current, the bead profile parameters increase linearly. The geometry of the bead profile was improved by using the GRA

The SDSS-IV extended Baryon Oscillation Spectroscopic Survey : Luminous Red Galaxy Target Selection

We describe the algorithm used to select the Luminous Red Galaxy (LRG) sample for the extended Baryon Oscillation Spectroscopic Survey (eBOSS) of the Sloan Digital Sky Survey IV (SDSS-IV) using photometric data from both the SDSS and the Wide-Field Infrared Survey Explorer (WISE). LRG targets are required to meet a set of color selection criteria and have z-band and i-band MODEL magnitudes z <19.95 and 19.9 <i < 21.8, respectively. Our algorithm selects roughly 50 LRG targets per square degree, the great majority of which lie in the redshift range 0.6 <z <1.0 (median redshift 0.71). We demonstrate that our methods are highly effective at eliminating stellar contamination and lower-redshift galaxies. We perform a number of tests using spectroscopic data from SDSS-III/BOSS to determine the redshift reliability of our target selection and its ability to meet the science requirements of eBOSS. The SDSS spectra are of high enough signal-to-noise ratio that at least 89% of the target sample yields secure redshift measurements. We also present tests of the uniformity and homogeneity of the sample, demonstrating that it should be clean enough for studies of the large-scale structure of the universe at higher redshifts than SDSS-III/BOSS LRGs reached.Publisher PDFPeer reviewe