Development of a sequence-tagged site (STS) marker for sex identification in the dioecious rattan species Calamus guruba Buch.-Ham.

Molecular Breeding - With the decrease of cost in genotyping, single nucleotide polymorphisms (SNPs) have gained wide acceptance because of their abundance, even distribution throughout the maize...     

Production and characterization of Monoclonal antibodies to bluetongue virus

In the present study, a total of 24 MAbs were produced against bluetongue virus (BTV) by polyethyleneglycol (PEG) mediated fusion method using sensitized lymphocytes and myeloma cells. All these clones were characterized for their reactivity to whole virus and recombinant BTV-VP7 protein, titres, isotypes and their reactivity with 24 BTV-serotype specific sera in cELISA. Out of 24 clones, a majority of them (n = 18) belong to various IgG subclasses and the remaining (n = 6) to the IgM class. A panel of eight clones reactive to both whole BTV and purified rVP7 protein were identified based on their reactivity in iELISA. For competitive ELISA, the clone designated as 4A10 showed better inhibition to hyperimmune serum of BTV serotype 23. However, this clone showed a variable percent of inhibition ranging from16.6% with BTV 12 serotype to 78.9% with BTV16 serotype using 24 serotype specific sera of BTV originating from guinea pig at their lowest dilutions. From the available panel of clones, only 4A10 was found to have a possible diagnostic application.     

Dynamic model for predicting survival of mature larvae of Tribolium confusum during facility heat treatments

Structural heat treatment, a viable alternative to methyl bromide fumigation, involves raising the ambient temperature of food-processing facilities between 50 and 60 degrees C by using gas, electric, or steam heaters, and holding these elevated temperatures for 24 h or longer to kill stored-product insects. A dynamic model was developed to predict survival of mature larvae, which is the most heat-tolerant stage of the confused flour beetle, Tribolium confusum (Jacquelin du Val), at elevated temperatures between 46 and 60 degrees C. The model is based on two nonlinear relationships: 1) logarithmic survival of T. confusum mature larvae as a function of time, and 2) logarithmic reduction in larval survival as a function of temperature. The dynamic model was validated with nine independent data sets collected during actual facility heat treatments conducted on two separate occasions at the Kansas State University pilot flour and feed mills. The rate of increase of temperature over time varied among the nine locations where mature larvae of T. confusum were exposed, and the approximate heating rates during the entire heat treatment ranged from 1.1 to 13.2 degrees C/h. The absolute deviation in the predicted number of larvae surviving the heat treatment was within 3-7% of the actual observed data. Comparison of the absolute deviation in the time taken for equivalent larval survival showed that the model predictions were within 2-6% of the observed data. The dynamic model can be used to predict survival of mature larvae of T. confusum during heat treatments of food-processing facilities based on time-dependent temperature profiles obtained at any given location.     

Dual roles of immunoregulatory cytokine TGF-beta in the pathogenesis of autoimmunity-mediated organ damage

Ample evidence suggests a role of TGF-beta in preventing autoimmunity. Multiorgan inflammatory disease, spontaneous activation of self-reactive T cells, and autoantibody production are hallmarks of autoimmune diseases, such as lupus. These features are reminiscent of the immunopathology manifest in TGF-beta1-deficient mice. In this study, we show that lupus-prone (New Zealand Black and White)F(1) mice have reduced expression of TGF-beta1 in lymphoid tissues, and TGF-beta1 or TGF-beta1-producing T cells suppress autoantibody production. In contrast, the expression of TGF-beta1 protein and mRNA and TGF-beta signaling proteins (TGF-beta receptor type II and phosphorylated SMAD3) increases in the target organs, i.e., kidneys, of these mice as they age and develop progressive organ damage. In fact, the levels of TGF-beta1 in kidney tissue and urine correlate with the extent of chronic lesions that represent local tissue fibrosis. In vivo TGF-beta blockade by treatment of these mice with an anti-TGF-beta Ab selectively inhibits chronic fibrotic lesions without affecting autoantibody production and the inflammatory component of tissue injury. Thus, TGF-beta plays a dual, seemingly paradoxical, role in the development of organ damage in multiorgan autoimmune diseases. According to our working model, reduced TGF-beta in immune cells predisposes to immune dysregulation and autoantibody production, which causes tissue inflammation that triggers the production of anti-inflammatory cytokines such as TGF-beta in target organs to counter inflammation. Enhanced TGF-beta in target organs, in turn, can lead to dysregulated tissue repair, progressive fibrogenesis, and eventual end-organ damage.     

Differences in calmodulin and calmodulin-binding proteins in phasic and tonic smooth muscles

To determine whether densities ofcalmodulin (CaM) and CaM-binding proteins are related to phasic andtonic behavior of smooth muscles, we quantified these proteins in theopossum esophageal body (EB) and lower esophageal sphincter (LES),which represent phasic and tonic smooth muscles, respectively. Gelelectrophoresis, immunoprecipitation, Western blot, and hemagglutininepitope-tagged CaM (HA-CaM) overlay assay with quantitative scanningdensitometry and phosphorylation measurements were used. Total proteincontent in the two smooth muscles was similar (~30 mg protein/gfrozen tissue). Total tissue concentration of CaM was significantly(25%) higher in EB than in LES (P < 0.05).HA-CaM-binding proteins were qualitatively similar in LES and EBextracts. Myosin, myristoylated alanine-rich C kinase substrateprotein, Ca²⁺/CaM kinase II, and calponin contents werealso similar in the two muscles. However, content and total activity ofmyosin light chain kinase (MLCK) and content of caldesmon (CaD) werethree- to fourfold higher in EB than in LES. Increased CaM and MLCKcontent may allow for a wide range of contractile force varying fromcomplete relaxation in the basal state to a large-amplitude,high-velocity contraction in EB phasic muscle. Increased content ofCaD, which provides a braking mechanism on contraction, may furthercontribute to the phasic contractile behavior. In contrast, low CaM,MLCK, and CaD content may be responsible for a small range ofcontractile force seen in tonic muscle of LES.

     

Toxicity and disruption of midgut physiology in larvae of the European corn borer,Ostrinia nubilalis,by anion transporter blockers

In this study, four blockers of anion transporters (ATs) belonging to four different classes of organic acids, including DIDS (4, 4'‐diisothiocyanatostilbene‐2, 2'‐ disulfonic acid; a stilbene disulfonic acid), NPPB [(5‐nitro‐2‐(3‐phenylpropylamino) benzoic acid; an anthranilic acid)], 9‐AC (anthracene‐9‐carboxylic acid; an aromatic carboxylic acid), and IAA‐94 (indanyloxy acetic acid; an indanyloxy alkanoic acid), were tested for their toxicity against the European corn borer (ECB), Ostrinia nubilalis. All the AT blockers inhibited the growth of larvae, increased the developmental time, and decreased survival compared to controls, when second‐instar ECB larvae were fed for seven days on treated diet. In general, DIDS and NPPB were the most active compounds, with the rank order of activity being DIDS>NPPB>IAA‐94>9‐AC. All the AT blockers decreased the midgut alkalinity in fifth‐instar larvae when fed for 3 h on treated diet. Effective concentrations required for 50% decrease in midgut alkalinity (EC₅₀) ranged between 29.1 and 41.2 ppm and the rank order of activity was NPPB>DIDS>IAA‐94>9‐AC. Similarly, all the tested AT blockers inhibited ³⁶Cl^? uptake from the midgut lumen in fifth‐instar larvae when fed for 3 h on treated diet. Concentrations required for 50% inhibition of ³⁶Cl^? uptake (IC₅₀) ranged between 7.4 and 11.0 ppm and the rank order of activity was DIDS>NPPB>9‐AC >IAA‐94. Modest to highly strong positive correlations observed among growth, midgut alkalinity, and midgut Cl^? ion transport in AT blocker–fed larvae suggested that these effects are causally related to each other. Finally, AT blockers have the potential to become good candidates for development of insecticides with a unique mode of action. © 2009 Wiley Periodicals, Inc.     

Performance of an anaerobic baffled reactor and hybrid constructed wetland treating high-strength wastewater in Nepal—A model for DEWATS

Centralized wastewater treatment systems require sophisticated technologies and skilled manpower for their operation and maintenance (O&M). These systems have huge construction as well as O&M costs. Therefore, a Decentralized Wastewater Treatment System (DEWATS) rather than a centralized system might be especially beneficial in developing countries. A model for DEWATS is developed in Nepal with Anaerobic Baffled Reactor (ABR) and hybrid Constructed Wetland (CW). The DEWATS treats high-strength wastewater from 80 households (400 PE). This paper summarizes the performance of the DEWATS from July 2006 to August 2007 in the removal efficiencies of TSS, BOD₅, COD, NH₄–N, TP and FC. The ABR is very effective in the removal of organic pollutants and could achieve TSS removal up to 91%, BOD₅ up to 78% and COD up to 77%. The average removal efficiencies of the DEWATS is 96% TSS, 90% BOD₅, 90% COD, 70% NH₄–N, 26% TP and 98% FC.