Antioxidant responses and water status in Brassica seedlings subjected to boron stress

The effect of boron (B) on growth, water status and oxidative damage was investigated in the leaves and roots of 7-day-old seedlings of Brassica juncea var. Varuna. For this seedlings of Brassica were grown in solution culture with variable boron supply (0.033, 0.33, 3.3 and 33 mg B L^?1) under controlled conditions in green house. Photosynthetic pigments were found to be decreased more under excess (3.3 and 33 mg B L^–1) than deficient boron supply (0.033 mg B L^–1) when compared to control (0.33 mg B L^–1). Accumulation of hydrogen peroxide and thiobarbituric acid reactive substances content in both leaves and roots under deficient and excess boron supply suggested oxidative damage due to excessive production of reactive oxygen species. Increased activity of antioxidative enzymes: superoxide dismutase, catalase and peroxidase along with polyphenol oxidase was observed in leaves and roots under boron deficiency and excess than in control. Increased proline concentration, decreased total water content and water saturation deficit also indicated the water deficit condition in leaves and roots of boron-stressed Brassica seedlings.     

The impact of foliar boron sprays on reproductive biology and seed quality of black gram

An experiment was conducted under glass house condition to study the effect of foliar application of boron (B) on reproductive biology and seed quality of black gram (Vigna mungo). Black gram (V. mungo L. var. DPU-88-31) was grown under controlled sand culture condition at deficient and sufficient B levels. After 32 days of sowing B deficient plants were sprayed with three concentrations of B (0.05%, 0.1% and 0.2% borax) at three different stages of reproductive development, i.e. prior to flowering, initiation of bud formation and after bud formation. Deficient B supply decreased the anther and pollen size, pollen tube growth, pollen viability as well as stigmatic receptivity which were increased by foliar B application. Foliar spray at all the three concentrations and at all stages increased the yield parameters like number of pods, pod size and number of seeds formed per plant. Foliar B application also improved the seed yield and seed quality in terms of storage seed proteins (albumin, globulin, glutenin and prolamin) and carbohydrates (sugars and starch) in black gram. The foliar application of B in appropriate doses (particularly 0.1%) after bud formation made quantitative and qualitative improvement in seed yield of black gram by supplementing additional/critical B requirements for reproductive development.     

The cytosolic chaperone Hsc70 promotes traffic to the cell surface of intracellular retained melanocortin-4 receptor mutants

Inherited modifications in protein structure frequently cause a loss-of-function by interfering with protein synthesis, transport, or stability. For the obesity-linked melanocortin-4 receptor (MC4R) and other G protein-coupled receptors, many mutants are intracellular retained. The biogenesis and trafficking of G protein-coupled receptors are regulated by multiple factors, including molecular chaperone networks. Here, we have investigated the ability of the cytosolic cognate 70-kDa heat-shock protein (Hsc70) chaperone system to modulate cell surface expression of MC4R. Clinically occurring MC4R mutants S58C, P78L, and D90N were demonstrated to have reduced trafficking to the plasma membrane and to be retained at the endoplasmic reticulum (ER). Analyses by fluorescence recovery after photobleaching revealed that the mobility of MC4R mutant protein at the ER was reduced, implying protein misfolding. In cells expressing MC4R, overexpression of Hsc70 resulted in increased levels of wild-type and mutant receptors at the cell surface. MC4R and Hsc70 coimmunoprecipitated, and fluorescence recovery after photobleaching analyses showed that increasing cellular levels of Hsc70 promoted the mobility of ER retained MC4R. Moreover, expression of HSJ1b, a cochaperone that enhances degradation of Hsc70 clients, reduced cellular levels of MC4R. Hsp70 and Hsp90 chaperone systems collaborate in the cellular processing of clients. For MC4R, inhibition of endogenous Hsp90 by geldanamycin reduced receptor levels. By contrast, expression of the Hsp90 cochaperone Aha1 (activator of Hsp90 ATPase) increased cellular levels of MC4R. Finally, we demonstrate that signaling of intracellular retained MC4R mutants is increased in cells overexpressing Hsc70. These data indicate that cytosolic chaperone systems can facilitate rescue of intracellular retained MC4R by improving folding. They also support proteostasis networks as a potential target for MC4R-linked obesity.     

CD59, a complement regulatory protein, controls choroidal neovascularization in a mouse model of wet-type age-related macular degeneration

We have shown that membrane attack complex (MAC) formation via the activation of the alternative pathway plays a central role in the laser-induced choroidal neovascularization (CNV). This study was undertaken to understand the role of a complement regulatory protein, CD59, which controls MAC assembly and function, in this model. CNV was induced by laser photocoagulation in C57BL/6 and Cd59a(-/-) mice using an argon laser. Animals from each group were sacrificed on day 1, 3, 5, and 7 postlaser. Retinal pigment epithelium-choroid-scleral tissue was examined to determine the incidence and size of CNV complex, and semiquantitative RT-PCR and Western blot analysis for CD59a was studied. Recombinant soluble mouse CD59a-IgG2a fusion (rsCD59a-Fc) protein was injected via i.p. or intravitreal routes 24 h before laser. Our results demonstrated that CD59a (both mRNA and protein) was down-regulated during laser-induced CNV. Cd59a(-/-) mice developed CNV complex early in the disease process. Increased MAC deposition was also observed in these Cd59a(-/-) mice. Administration of rsCD59a-Fc inhibited the development of CNV complex in the mouse model by blocking MAC formation and also inhibited expression of angiogenic growth factors. These data provide strong evidence that CD59a plays a crucial role in regulating complement activation and MAC formation essential for the release of growth factors that drive the development of laser-induced CNV in mice. Thus, our results suggest that the inhibition of complement by soluble CD59 may provide a novel therapeutic alternative to current treatment.     

Neutrophilic phagocytosis by tumor cells. A feature to consider in the differential diagnosis in fine needle aspiration smears

OBJECTIVE: To analyze neutrophilic phagocytosis by tumor cells in fine needle aspirate (FNA) smears from different types of tumor. STUDY DESIGN: A retrospective review of a total of 7 cases showing prominent neutrophilic phagocytosis by tumor cells in FNA smears during the period July 2003-December 2004. RESULTS: This feature was seen in malignant fibrous histiocytoma and poorly differentiated renal cell carcinoma in addition to giant cell carcinoma of the lung. CONCLUSION: Neutrophilic phagocytosis by tumor cells is seen in FNA smears and on cytomorphology. The differential diagnoses should include both pleomorphic sarcomas and carcinomas.