Starvation Resistance in a Stenothermal Species from the Indian Subcontinent: Mechanistic Basis of Clinal Variation

Parallel clines for starvation resistance and lipid content are well documented among drosophilids on the Indian subcontinent. However, the mechanistic basis of these clines has not been investigated so far. Here, we investigate the utilization of lipids during starvation as a function of duration of stress in D. ananassae. We found higher lipid content responsible for high starvation resistance at lower latitudes. Lipids were utilized during starvation only; not during any other climatic stresses like desiccation or thermal stresses. We also found a cline for consumption of total body lipids; as more content (out of total amount of lipids) was utilized by flies at lower latitudes and lesser at higher latitudes. But, there was no latitudinal cline for threshold lipid amount in the case of females while for males there was a positive cline. Lastly, parallel clines have evolved under contrasting climatic conditions i.e. drier and colder northern localities have flies with lower lipid and reduced starvation resistance while hot and humid localities favor flies with higher lipid levels and greater starvation tolerance. Thus, the evolution of clines associated with starvation and lipid content might have resulted due to specific ecological conditions i.e. humidity gradient on the Indian subcontinent.     

The FBXO4 Tumor Suppressor Functions as a Barrier to BrafV600E-Dependent Metastatic Melanoma

Cyclin D1–cyclin-dependent kinase 4/6 (CDK4/6) dysregulation is a major contributor to melanomagenesis. Clinical evidence has revealed that p16^INK4A, an allosteric inhibitor of CDK4/6, is inactivated in over half of human melanomas, and numerous animal models have demonstrated that p16^INK4A deletion promotes melanoma. FBXO4, a specificity factor for the E3 ligase that directs timely cyclin D1 proteolysis, has not been studied in melanoma. We demonstrate that Fbxo4 deficiency induces Braf-driven melanoma and that this phenotype depends on cyclin D1 accumulation in mice, underscoring the importance of this ubiquitin ligase in tumor suppression. Furthermore, we have identified a substrate-binding mutation, FBXO4 I377M, that selectively disrupts cyclin D1 degradation while preserving proteolysis of the other known FBXO4 substrate, TRF1. The I377M mutation and Fbxo4 deficiency result in nuclear accumulation of cyclin D1, a key transforming neoplastic event. Collectively, these data provide evidence that FBXO4 dysfunction, as a mechanism for cyclin D1 overexpression, is a contributor to human malignancy.     

Divergence for tolerance to thermal-stress related traits in two Drosophila species of immigrans group

We investigated associations of the species specific divergence of thermal adaptations of Drosophila nasuta and Drosophila immigrans with seasonal as well as geographical changes in the relative abundance. In D. immigrans, cold resistance and egg-to-adult viability after a cold-shock have shown a positive cline with latitude, whereas no such clinal patterns were evident for heat resistance as well as egg-to-adult viability after heat stress. In contrast, basal heat tolerance and egg-to-adult viability after a heat-shock increased towards the equator i.e. from north to south in D. nasuta. Further, we also found species-specific divergence in seasonal climatic selection responses for heat as well as cold resistance i.e. warmer and higher humidity conditions of the rainy season have evidenced ∼2-fold higher heat tolerance of D. nasuta, but lower temperature as well as lower humidity conditions of the autumn significantly increased (∼3-fold) cold tolerance of D. immigrans. Geographical variations were significant for the absolute and relative hardening capacity for cold tolerance in D. immigrans, but for heat resistance of D. nasuta. Results of multiple regression analysis of changes in thermal tolerance level in latitudinal populations as a simultaneous function of climatic variables (T_ave and RH) have shown that average yearly relative humidity levels significantly affect heat tolerance of D. nasuta, but both T_ave and RH contribute to changes in the cold tolerance of D. immigrans. Thus, our results suggest that climatic selection responses for basal and hardened thermo-tolerance levels affect relative abundance of D. nasuta and D. immigrans across a latitudinal transect as well as seasons.     

Genetic diversity within the genus Pleurotus determined by random amplified polymorphism DNA (RAPD) analysis

Random amplified polymorphism DNA (RAPD) analysis was done to assess the diversity among 10 species of Pleurotus. Understanding of the pattern not only addresses questions concerning evolutionary process and the development of conservation strategies, but also a pre-requisite of the efficient use of genetic resources in breeding programme. The RAPD dendogram obtained by using the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) programme were grouped into the investigated strains into five clusters. RAPD bands were scored as present (1) or absent (0) for all the Pleurotus isolate. Each band was assumed to represent a unique genetic locus. The pattern and extent of RAPD variation were analysed with respect to primer, polymorphic locus and isolate. Total number of amplified fragment and polymorphic fragment produced by 40 decamer primer was 229 and 226, respectively. Polymorphism percentage was 98.69. Ten primers; SBSA11, SBSA13, SBSA15, SBSA16, SBSA18, SBSA19, SBSA20, SBSB14, SBSB15 and SBSB17 were not amplified to the DNA from any of the isolate.     

Ceramide 1-phosphate stimulates glucose uptake in macrophages

It is well established that ceramide 1-phosphate (C1P) is mitogenic and antiapoptotic, and that it is implicated in the regulation of macrophage migration. These activities require high energy levels to be available in cells. Macrophages obtain most of their energy from glucose. In this work, we demonstrate that C1P enhances glucose uptake in RAW264.7 macrophages. The major glucose transporter involved in this action was found to be GLUT 3, as determined by measuring its translocation from the cytosol to the plasma membrane. C1P-stimulated glucose uptake was blocked by selective inhibitors of phosphatidylinositol 3-kinase (PI3K) or Akt, also known as protein kinase B (PKB), and by specific siRNAs to silence the genes encoding for these kinases. C1P-stimulated glucose uptake was also inhibited by pertussis toxin (PTX) and by the siRNA that inhibited GLUT 3 expression. C1P increased the affinity of the glucose transporter for its substrate, and enhanced glucose metabolism to produce ATP. The latter action was also inhibited by PI3K- and Akt-selective inhibitors, PTX, or by specific siRNAs to inhibit GLUT 3 expression.     

Isolation of proline-based cyclic dipeptides from Bacillus sp. N strain associated with rhabitid entomopathogenic nematode and its antimicrobial properties

Entomopathogenic nematodes (EPN) are well-known as biological control agents and are found to have associated bacteria which can produce a wide range of bioactive secondary metabolites. We report herewith isolation of six proline containing cyclic dipeptides cyclo(d-Pro-l-Leu), cyclo(l-Pro-l-Met), cyclo(d-Pro-l-Phe), cyclo(l-Pro-l-Phe), cyclo(l-Pro-l-Tyr) and cyclo(l-Pro-d-Tyr) from ethyl acetate extract of the Luria Broth (LB) cell free culture filtrate of Bacillus sp. strain N associated with a new EPN Rhabditis sp. from sweet potato weevil grubs collected from Central Tuber Crops Research Institute farm. Antimicrobial studies of these 2,5-diketopiperazines (DKPs) against both medicinally and agriculturally important bacterium and fungi showed potent inhibitory values in the range of μg/mL. Cyclic dipeptides showed significantly higher activity than the commercial fungicide bavistin against agriculturally important fungi, viz., Fusarium oxysporum, Rhizoctonia solani, and Pencillium expansum. The highest activity of 2 μg/mL by cyclo(l-Pro-l-Phe) was recorded against P. expansum, a plant pathogen responsible for causing post harvest decay of stored apples and oranges. To our knowledge, this is the first report on the isolation of these DKPs from Rhabditis EPN bacterial strain Bacillus sp.