Transforming genes among three different oncogenic subgroups of human adenoviruses have similar replicative functions.

We have examined the functional similarity of the transforming genes for replicative functions among three different subgroups of human adenoviruses (A, B, and C), using mutant complementation as an assay. A host range deletion mutant (dl201.2) of Ad2 (nononcogenic subgroup C) lacking about 5% of the viral DNA covering two early gene blocks (E1a and E1b) involved in cellular transformation was isolated and tested for its ability to replicate in nonpermissive KB cells in the presence of Ad7 (weakly oncogenic group B) or ad12 (highly oncogenic group A). The complementation of the mutant defect was demonstrated by cleaving the viral DNA extracted from mixed infected cells or the DNA extracted from purified virions from mixed infected cells with restriction endonuclease BamHI, which produces a different cleavage pattern with the DNA of each serotype. It was found that the defects in E1a plus E1b of dl201.2 could be complemented by Ad7 and Ad12, indicating that these genes in Ad2, Ad7, and Ad12 have similar functions during productive infection.     

A unique subgenomic species of adenovirus 2 DNA generated under high multiplicities of infection.

We have identified a novel subgenomic viral DNA in KB cells infected with adenovirus 2 (Ad2) under high multiplicities of infection. KB cells were infected with Ad2 at multiplicities of infection greater than 100 PFU/cell. 32P-labeled viral DNA was selectively extracted by a modification of the method of Hirt (8) from the infected cells and analyzed by electrophoresis on agarose gels. In addition to full-length DNA (33 to 23 x 10(6) daltons), a unique subgenomic DNA species of about 12 to 13% (2.6 x 10(6) daltons) of full-length DNA in size was found in the infected cells. This subgenomic DNA was found to be double stranded and was not packaged inside the virus particles. This DNA could be isolated in large amounts (30 to 50% of total viral DNA) from infected cells. When cleaved with restriction endonuclease KpnI, the subgenomic DNA yielded two fragments, each corresponding to about 6% and 7% of the full-length genome in size.     

Length weight relationship of five deep sea fishes from Kerala,south west coast of India

Length–weight relationships (LWRs) were estimated for five deep sea fishes viz. Astronesthes martensii, Glyptophidium macropus, Neobythites multistriatus, Physiculus roseus, Synagrops japonicus from Kerala, south west coast of India. Fishes were collected from commercial trawlers monthly from February 2018 to March 2019 operating at depth ranged from 270 m (Lat. 9°29.35′ N, Long. 75°44.74′ E) to 350 m (Lat. 9°26. 49′ N, Long. 75°42.36′ E) in the south east Arabian Sea. Correlation coefficients (r²) were found high for all species, with b value ranged from 2.923 to 3.404.     

Phosphorylation of the pro-apoptotic protein BIK: mapping of phosphorylation sites and effect on apoptosis

BIK is a pro-apoptotic BCL-2 family member and is the founding member of a subfamily of pro-apoptotic proteins known as "BH3-alone" proteins. Ectopic expression of BIK induces apoptosis in variety of mammalian cells. BIK complexes with various anti-apoptotic BCL-2 family proteins such as adenovirus E1B-19K and BCL-2 via the BH3 domain. However, the heterodimerization activity of BIK alone is insufficient for its apoptotic activity. Previous studies have shown that phosphorylation regulates the functional activity of both anti-apoptotic and pro-apoptotic members of the BCL-2 family. Here, we have examined phosphorylation of BIK and its effect on the apoptotic activity of BIK. We show that BIK exists as a phosphoprotein and is phosphorylated at residues 33 (threonine) and 35 (serine). Mutation of the phosphorylation sites, in which the Thr and Ser residues were changed to alanine residues, reduced the apoptotic activity of BIK without significantly affecting its ability to heterodimerize with BCL-2. Our results suggest that phosphorylation of BIK is required for eliciting efficient apoptotic activity. Partial purification of the protein kinase from HeLa cell cytoplasmic extracts suggest that BIK may be phosphorylated by a casein kinase II-related enzyme.     

Requirement of BAX for efficient adenovirus-induced apoptosis

Infection of human epithelial cells with adenoviruses induces an apoptosis paradigm that is efficiently suppressed by the expression of viral E1B-19K protein, which is a functional homolog of the cellular antiapoptosis protein BCL-2. The mechanisms of adenovirus (Ad)-induced apoptosis appear to involve the cellular BCL-2 family proapoptotic proteins. Recent genetic studies with fibroblasts derived from mutant mouse embryos indicate that a class of the BCL-2 family proapoptotic proteins (designated BH-123 or multidomain proteins) such as BAX and BAK constitutes an essential component of the core apoptosis machinery in animal cells. We have examined the role of BAX in Ad-induced apoptosis in human epithelial cells using two colon cancer cell lines, HCT116Bax (Bax(+/-)) and HCT116BaxKO (Bax(-/-)) (L. Zhang, J. Yu, B. H. Park, K. W. Kinzler, and B. Vogelstein, Science 290:989-992, 2000). Infection of Bax(+/-) cells with an Ad type 2 mutant (dl250) defective in expression of the E1B-19K protein resulted in enhanced cytopathic effect, large plaques on cell monolayers, fragmentation of cellular DNA, and enhanced cell death. These mutant phenotypes were not efficiently expressed in Bax(-/-) cells, suggesting that BAX is essential for Ad-induced apoptosis. Infection of Bax(+/-) cells with dl250 induced increased levels of an N-terminally processed form of BAX. Cells infected with the 19K mutant also contained enhanced levels of truncated BAX in membrane-inserted form. Our results suggest that at least a part of the mechanism utilized by E1B-19K to suppress apoptosis during Ad infection may involve modulation of the activities of BAX.     

Batch and continuous removal of arsenic using hyacinth roots

Arsenic is considered a primary pollutant in drinking water because of its high toxicity. The unique property of water hyacinth roots (Eichhornia crassipes) to remove heavy metals is of great signiicance for the development of a cost-effective phytoremediation technology. An experimental test program was conducted at the United States Environmental Protection (USEPA) Test and Evaluation (T&E) Facility in Cincinnati, Ohio, to investigate the potential of water hyacinth roots to remove arsenic from spiked drinking water samples. Water hyacinth roots were washed, dried, and powdered to provide dried hyacinth roots (DHR) for batch and continuous column experiments, Various quantities of DHR were added to water spiked with 300 micrograms per liter (microg/L) arsenic. A concentration of 20 g/L DHR was found adequate for greater than 90% arsenic removal in the batch tests. Based on the batch test results, continuous column experiments were performed using a 2-L column. In a continuous system, 15 L of water containing 300 microg/L arsenic were treated to below 20 microg/L using 50 g DHR, and 44 L of water containing 600 microg/L arsenic were treated to below 20 microg/L using 100 g DHR, giving a specific accumulation rate of approximately 260 microg As/g DHR.     

A Comparative Study on Torque Requirement During Extrusion Pretreatment of Different Feedstocks

Extrusion pretreatment of biomass can be one of the viable continuous pretreatment methods. The torque requirement of feedstock during extrusion was an important factor, and it was not reported in the literature. Screw compression ratio, screw speed, barrel temperature, and feedstock moisture content are the contributing factors to the torque. The current study was undertaken to investigate the effect of screw compression ratio, screw speed, temperature on torque requirement for different moisture content of switchgrass, prairie cord grass, corn stover, and big bluestem and to compare the torque requirement among the selected feedstocks. Biomass was extruded in a lab scale single-screw extruder with different screw compression ratios (2:1 and 3:1), screw speeds (50, 100, and 150?rpm), and barrel temperatures (50°C, 100°C, and 150°C) over a range of moisture contents (15%, 25%, 35%, and 45% wb). Statistical analyses revealed that all the independent variables considered in this study had a significant effect on torque requirement for the selected feedstocks. Among the independent variables considered moisture content, screw speed, and temperature had a negative effect on torque requirement for all the feedstocks. Switchgrass required the highest torque followed by corn stover, big bluestem, and prairie cord grass.     

Cooperativity between inhibition of cytosolic K+ efflux and AMPK activation during suppression of hypoxia-induced cellular apoptosis

Cellular potassium homeostasis has recently emerged as a critical regulator of apoptosis in response to variety of stimuli. However, functional hierarchy of this phenomenon in the apoptotic cascade and therefore, its significance as a pathway for intervention is not fully established. Chronic hypoxia, a known threat to cell survival, also modulates cellular potassium homeostasis. In this study, we tested if hypoxia-induced apoptosis in lymphocytes can be prevented by modulating cellular K+ homeostasis. We observed that chronic hypoxia accelerated the rate of apoptosis in resting murine splenocytes concomitant with cytosolic K+ efflux. We tested several modalities including elevated extracellular potassium besides various K+ channel inhibitors to curtail hypoxia-induced K+ efflux and interestingly, established that the supplementation of KCl in extracellular medium was most effective in preventing hypoxia-induced apoptosis in these cells. Subsequent mechanistic dissection of pathways underlying this phenomenon revealed that besides effectively inhibiting hypoxia-induced efflux of K+ ion and its downstream cell-physiological consequences; elevated extracellular KCl modulated steady state levels of cellular ATP and culminated in stabilization of AMPKα with pro-survival consequences. Also, interestingly, global gene expression profiling revealed that KCl supplementation down regulated a distinct p53-regulated cellular sub-network of genes involved in regulation of DNA replication. Additionally, we present experimental evidence for the functional role of AMPK and p53 activation during suppression of hypoxia-induced apoptosis. In conclusion, our study highlights a novel bimodal effect wherein cooperativity between restoration of K+ homeostasis and a sustainable ‘metabolic quiescence’ induced by AMPK activation appeared indispensible for curtailing hypoxia-induced apoptosis.