Screening of new isolates of Bacillus thuringiensis (Bt) and cloning of the cry genes

Nine new indigenous isolates of Bacillus thuringiensis (Bt) were characterized for their colony type, crystal inclusion and toxicity analysis with Helicoverpa armigera Hubner and Spodoptera litura Linn. Genomic deoxyribonucleic acid (DNA) isolated from all the new isolates were subjected to screening for cry1, cry2, cry4, cry10 and cry11 genes and predicted possible potential DNA amplicons were cloned and sequenced. Partial cry1 gene fragment (~1.5 kb) amplified by degenerate primers and about 450 bp DNA fragment amplified by cry10 gene specific primers from two isolates T109 and T136 were cloned in to T/A cloning vector. DNA sequencing of about 1.5 kb amplicon showed 99% homology to the holotype sequence of cry1Ac1. Nucleotide sequence of about 450 bp fragments of isolate T109 and T136 showed homology to a hypothetical protein and serine/threonine phosphatase respectively.Keywords: Bacillus thuringiensis (Bt), cloning, cry genes, polymerase chain reaction (PCR), toxicity analysi

Silkworm Thermal Biology: A Review of Heat Shock Response, Heat Shock Proteins and Heat Acclimation in the Domesticated Silkworm, Bombyx mori

Heat shock proteins (HSPs) are known to play ecological and evolutionary roles in this postgenomic era. Recent research suggests that HSPs are implicated in cardiovascular biology and disease development, proliferation and regulation of cancer cells, cell death via apoptosis, and several other key cellular functions. These activities have generated great interest amongst cell and molecular biologists, and these biologists are keen to unravel other hitherto unknown potential functions of this group of proteins. Consequently, the biological significance of HSPs has led to cloning and characterization of genes encoding HSPs in many organisms including the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). However, most of the past investigations in B. mori were confined to expression of HSPs in tissues and cell lines, whereas information on their specific functional roles in biological, physiological, and molecular processes is scarce. Naturally occurring or domesticated polyvoltines (known to be the tropical race) are more resistant to high temperatures and diseases than bi- or univoltines (temperate races). The mechanism of ecological or evolutionary modification of HSPs during the course of domestication of B. mori - particularly in relation to thermotolerance in geographically distinct races/strains - is still unclear. In addition, the heat shock response, thermal acclimation, and hardening have not been studied extensively in B. mori compared to other organisms. Towards this, recent investigations on differential expression of HSPs at various stages of development, considering the concept of the whole organism, open ample scope to evaluate their biological and commercial importance in B. mori which has not been addressed in any of the representative organisms studied so far. Comparatively, heat shock response among different silkworm races/strains of poly-, bi-, and univoltines varies significantly and thermotolerance increases as the larval development proceeds. Hence, this being the first review in this area, an attempt has been made to collate all available information on the heat shock response, HSPs expression, associated genes, amino acid sequences, and acquired/unacquired thermotolerance. The aim is to present this as a valuable resource for addressing the gap in knowledge and understanding evolutionary significance of HSPs between domesticated (B. mori) and non-domesticated insects. It is believed that the information presented here will also help researchers/breeders to design appropriate strategies for developing novel strains for the tropics

Conical Defects in Higher Spin Theories

We study conical defect geometries in the SL(N) Chern-Simons formulation of higher spin gauge theories in AdS_3. We argue that (for N\geq 4) there are special values of the deficit angle for which these geometries are actually smooth configurations of the underlying theory. We also exhibit a gauge in which these geometries can be viewed as wormholes interpolating between two distinct asymptotically AdS_3 spacetimes. Remarkably, the spectrum of smooth SL(N,C) solutions, after an appropriate analytic continuation, exactly matches that of the so-called "light primaries" in the minimal model W_N CFTs at finite N. This gives a candidate bulk interpretation of the latter states in the holographic duality proposed in [1].Comment: 38 page

Role for substance p-based nociceptive signaling in progenitor cell activation and angiogenesis during ischemia in mice and in human subjects

Our data highlight the role of SP in reparative neovascularization. Nociceptive signaling may represent a novel target of regenerative medicine

Tetraspanin 6: a pivotal protein of the multiple vesicular body determining exosome release and lysosomal degradation of amyloid precursor protein fragments

BACKGROUND: The mechanisms behind Aβ-peptide accumulation in non-familial Alzheimer’s disease (AD) remain elusive. Proteins of the tetraspanin family modulate Aβ production by interacting to γ-secretase. METHODS: We searched for tetraspanins with altered expression in AD brains. The function of the selected tetraspanin was studied in vitro and the physiological relevance of our findings was confirmed in vivo. RESULTS: Tetraspanin-6 (TSPAN6) is increased in AD brains and overexpression in cells exerts paradoxical effects on Amyloid Precursor Protein (APP) metabolism, increasing APP-C-terminal fragments (APP-CTF) and Aβ levels at the same time. TSPAN6 affects autophagosome-lysosomal fusion slowing down the degradation of APP-CTF. TSPAN6 recruits also the cytosolic, exosome-forming adaptor syntenin which increases secretion of exosomes that contain APP-CTF. CONCLUSIONS: TSPAN6 is a key player in the bifurcation between lysosomal-dependent degradation and exosome mediated secretion of APP-CTF. This corroborates the central role of the autophagosomal/lysosomal pathway in APP metabolism and shows that TSPAN6 is a crucial player in APP-CTF turnover

Quantum critical spin-liquid-like behavior in S = 1/2 quasi-kagome lattice compound CeRh₁-ₓPdₓSn investigated using muon spin relaxation and neutron scattering

We present the results of muon spin relaxation (μSR) and neutron scattering on the Ce-based quasikagome lattice CeRh1−xPdxSn (x=0.1 to 0.75). Our ZF-μSR results reveal the absence of static long-range magnetic order down to 0.05~K in x=0.1 single crystals. The weak temperature-dependent plateaus of the dynamic spin fluctuations below 0.2~K in ZF-μSR together with its longitudinal-field (LF) dependence between 0 and 3~kG indicate the presence of dynamic spin fluctuations persisting even at T = 0.05~K without static magnetic order. On the other hand, C4f/T increases as --log T on cooling below 0.9~K, passes through a broad maximum at 0.13~K and slightly decreases on further cooling. The ac-susceptibility also exhibits a frequency independent broad peak at 0.16~K, which is prominent with an applied field H along c-direction. We, therefore, argue that such a behavior for x=0.1 (namely, a plateau in spin relaxation rate (λ) below 0.2~K and a linear T dependence in C4f below 0.13~K) can be attributed to a metallic spin-liquid (SL) ground state near the quantum critical point in the frustrated Kondo lattice. The LF-μSR study suggests that the out of kagome plane spin fluctuations are responsible for the SL behavior. Low energy inelastic neutron scattering (INS) of x = 0.1 reveals gapless magnetic excitations, which are also supported by the behavior of C4f proportional to T1.1 down to 0.06~K

What are we measuring? A critique of range of motion methods currently in use for Dupuytren's disease and recommendations for practice

Background: Range of motion is the most frequently reported measure used in practice to evaluate outcomes. A goniometer is the most reliable tool to assess range of motion yet, the lack of consistency in reporting prevents comparison between studies. The aim of this study is to identify how range of motion is currently assessed and reported in Dupuytren’s disease literature. Following analysis recommendations for practice will be made to enable consistency in future studies for comparability. This paper highlights the variation in range of motion reporting in Dupuytren’s disease. Methods: A Participants, Intervention, Comparison, Outcomes and Study design format was used for the search strategy and search terms. Surgery, needle fasciotomy or collagenase injection for primary or recurrent Dupuytren’s disease in adults were included if outcomes were monitored using range of motion to record change. A literature search was performed in May 2013 using subject heading and free-text terms to also capture electronic publications ahead of print. In total 638 publications were identified and following screening 90 articles met the inclusion criteria. Data was extracted and entered onto a spreadsheet for analysis. A thematic analysis was carried out to establish any duplication, resulting in the final range of motion measures identified. Results: Range of motion measurement lacked clarity, with goniometry reportedly used in only 43 of the 90 studies, 16 stated the use of a range of motion protocol. A total of 24 different descriptors were identified describing range of motion in the 90 studies. While some studies reported active range of motion, others reported passive or were unclear. Eight of the 24 categories were identified through thematic analysis as possibly describing the same measure, ‘lack of joint extension’ and accounted for the most frequently used. Conclusions: Published studies lacked clarity in reporting range of motion, preventing data comparison and meta-analysis. Percentage change lacks context and without access to raw data, does not allow direct comparison of baseline characteristics. A clear description of what is being measured within each study was required. It is recommended that range of motion measuring and reporting for Dupuytren’s disease requires consistency to address issues that fall into 3 main categories:- Definition of terms Protocol statement Outcome reportin

The malaria candidate vaccine liver stage antigen-3 is highly conserved in Plasmodium falciparum isolates from diverse geographical areas

<p>Abstract</p> <p>Background</p> <p>A high level of genetic stability has been formerly identified in segments of the gene coding for the liver stage antigen-3 (LSA-3), a subunit vaccine candidate against <it>Plasmodium falciparum</it>. The exploration of <it>lsa-3 </it>polymorphisms was extended to the whole sequence of this large antigen in 20 clinical isolates from four geographical areas; Senegal, Comoro islands, Brazil and Thailand.</p> <p>Methods</p> <p>The whole 4680 bp genomic sequence of <it>lsa-3 </it>was amplified by polymerase chain reaction and sequenced. The clinical isolate sequences were aligned on the sequence of the laboratory reference <it>P. falciparum </it>strain 3D7.</p> <p>Results</p> <p>The non-repeated sequence of <it>lsa-3 </it>was very well conserved with only a few allelic variations scattered along the sequence. Interestingly, a formerly identified immunodominant region, employed for the majority of pre-clinical vaccine development, was totally conserved at the genetic level. The most significant variations observed were in the number and organization of tetrapeptide repeated units, but not in their composition, resulting in different lengths of these repeated regions. The shorter repeated regions were from Brazilian origin. A correlation between the geographical distribution of the parasites with single nucleotide polymorphisms was not detected.</p> <p>Conclusion</p> <p>The lack of correlation between allelic polymorphisms with a specific transmission pressure suggests that LSA-3 is a structurally constrained molecule. The unusual characteristics of the <it>lsa-3 </it>gene make the molecule an interesting candidate for a subunit vaccine against malaria.</p

Genomic Organization, Tissue Distribution and Functional Characterization of the Rat Pate Gene Cluster

The cysteine rich prostate and testis expressed (Pate) proteins identified till date are thought to resemble the three fingered protein/urokinase-type plasminogen activator receptor proteins. In this study, for the first time, we report the identification, cloning and characterization of rat Pate gene cluster and also determine the expression pattern. The rat Pate genes are clustered on chromosome 8 and their predicted proteins retained the ten cysteine signature characteristic to TFP/Ly-6 protein family. PATE and PATE-F three dimensional protein structure was found to be similar to that of the toxin bucandin. Though Pate gene expression is thought to be prostate and testis specific, we observed that rat Pate genes are also expressed in seminal vesicle and epididymis and in tissues beyond the male reproductive tract. In the developing rats (20–60 day old), expression of Pate genes seem to be androgen dependent in the epididymis and testis. In the adult rat, androgen ablation resulted in down regulation of the majority of Pate genes in the epididymides. PATE and PATE-F proteins were found to be expressed abundantly in the male reproductive tract of rats and on the sperm. Recombinant PATE protein exhibited potent antibacterial activity, whereas PATE-F did not exhibit any antibacterial activity. Pate expression was induced in the epididymides when challenged with LPS. Based on our results, we conclude that rat PATE proteins may contribute to the reproductive and defense functions

Gut-microbiota link in Parkinson’s disease: current perspectives

Parkinson’s disease(PD) is a metacentric neurodegenerative disorder results with accumulation and aggregation of alpha-synculein(α-Syn) (or alpha-synculeinopathy) in the substantia nigra in the central nervous system(CNS).Contributory factors include pesticide exposure, head injury and agriculture background. PD has been considered to be a non-genetic disorder, however around 15% individuals with PD have firstdegree relative who has the disease Mutations in genes including SNCA,LRRK2 and gluococerebrosidase (GBA) found to be risk factor for sporadic PD. Brain cells could be lost due to an abnormal accumulation of the protein alpha-synculein.This insoluble protein accumulates inside neurons forming inclusions called Lewy bodies. Other cell death mechanisms include proteasomal and lysosomal system dysfunction, but the mechanisms are not fully understood. Brain –gut axis(GBA) refers to central nervous system(CNS) control of the enteric nervous system(ENS) through vagus nerve intervention. PD is characterized by alphasynculeinopathy affecting all levels of the brain-gut axis.Both clinical and neuropathological evidences indicate the neurodegenerative changes in PD are accompanied by gastrointestinal symptoms that may precede or follow the central nervous systemimpairment. Frequent symptoms in PD include tremor, rigidity, slowness of movement and difficulty with walking. Treatment with L-DOPA(levodopa),with dopamine agonist, medications become less effective and produce complications. Research studies recommend new therapeutic approach in PD based on modification of the gut microbiota with probiotics, prebiotics, or even fecal microbiota transplantation