Survival response to increased ceramide involves metabolic adaptation through novel regulators of glycolysis and lipolysis.

The sphingolipid ceramide elicits several stress responses, however, organisms survive despite increased ceramide but how they do so is poorly understood. We demonstrate here that the AKT/FOXO pathway regulates survival in increased ceramide environment by metabolic adaptation involving changes in glycolysis and lipolysis through novel downstream targets. We show that ceramide kinase mutants accumulate ceramide and this leads to reduction in energy levels due to compromised oxidative phosphorylation. Mutants show increased activation of Akt and a consequent decrease in FOXO levels. These changes lead to enhanced glycolysis by upregulating the activity of phosphoglyceromutase, enolase, pyruvate kinase, and lactate dehydrogenase to provide energy. A second major consequence of AKT/FOXO reprogramming in the mutants is the increased mobilization of lipid from the gut through novel lipase targets, CG8093 and CG6277 for energy contribution. Ubiquitous reduction of these targets by knockdown experiments results in semi or total lethality of the mutants, demonstrating the importance of activating them. The efficiency of these adaptive mechanisms decreases with age and leads to reduction in adult life span of the mutants. In particular, mutants develop cardiac dysfunction with age, likely reflecting the high energy requirement of a well-functioning heart. The lipases also regulate physiological triacylglycerol homeostasis and are important for energy metabolism since midgut specific reduction of them in wild type flies results in increased sensitivity to starvation and accumulation of triglycerides leading to cardiac defects. The central findings of increased AKT activation, decreased FOXO level and activation of phosphoglyceromutase and pyruvate kinase are also observed in mice heterozygous for ceramide transfer protein suggesting a conserved role of this pathway in mammals. These data reveal novel glycolytic and non-autonomous lipolytic pathways in response to increased ceramide for sustenance of high energy demanding organ functions like the heart.

A nutrient responsive lipase mediates gut-brain communication to regulate insulin secretion in Drosophila.

Pancreatic ß cells secrete insulin in response to glucose elevation to maintain glucose homeostasis. A complex network of inter-organ communication operates to modulate insulin secretion and regulate glucose levels after a meal. Lipids obtained from diet or generated intracellularly are known to amplify glucose-stimulated insulin secretion, however, the underlying mechanisms are not completely understood. Here, we show that a Drosophila secretory lipase, Vaha (CG8093), is synthesized in the midgut and moves to the brain where it concentrates in the insulin-producing cells in a process requiring Lipid Transfer Particle, a lipoprotein originating in the fat body. In response to dietary fat, Vaha stimulates insulin-like peptide release (ILP), and Vaha deficiency results in reduced circulatory ILP and diabetic features including hyperglycemia and hyperlipidemia. Our findings suggest Vaha functions as a diacylglycerol lipase physiologically, by being a molecular link between dietary fat and lipid amplified insulin secretion in a gut-brain axis.

In Silico Characterization of Intracellular Localization Signals and Structural Features of Mosquito Densovirus (MDV) Viral Proteins.

As entomopathogenic viruses, mosquito densoviruses (MDVs) are widely studied for their potential as biocontrol agents and molecular laboratory tools for mosquito manipulation. The nucleus of the mosquito cell is the site for MDV genome replication and capsid assembly, however the nuclear localization signals (NLSs) and nuclear export signals (NES) for MDV proteins have not yet been identified. We carried out an in silico analysis to identify putative NLSs and NESs in the viral proteins of densoviruses that infect diverse mosquito genera (Aedes, Anopheles, and Culex) and identified putative phosphorylation and glycosylation sites on these proteins. These analyses lead to a more comprehensive understanding of how MDVs are transported into and out of the nucleus and lay the foundation for the potential use of densoviruses in mosquito control and basic research.

Effect of vitamins C and E on spermatogenesis in mice exposed to cadmium.

Cadmium (Cd) is a potential pollutant of the environment. It manifests cyto-toxic effects in different organs in animals. In the present study, intraperitoneal injection of CdCl(2) (1mg/kg body weight) increased lipid peroxidation in Swiss mice testes indicating oxidative stress during 5th to 8th week of post-treatment . The enzymatic activity of superoxide dismutase (SOD), catalase (CT) and peroxidase (PD) were significantly decreased over the post-treatment phase in Cd-treated mice testes compared to vehicle controls. Further, ascorbic acid content also declined significantly in Cd-exposed mice testes. Following Cd treatment, a marked increase in sperm abnormality percentage and significant decrease in sperm count was observed. The purpose of the present study was to evaluate the effect of vitamins C and E supplementation on Cd-treated mice testes. Therefore, Cd-treated mice groups were injected with vitamins C and E, separately, to assess the effect of the vitamins in combating Cd-induced cytotoxicity and other manifestations. Supplementation of vitamin C (10mg/kg body weight) and vitamin E (100mg/kg body weight) to Cd-induced mice groups declined lipid peroxidation, increased sperm count profile, depressed the percentage of sperm abnormality, increased the activity of antioxidant enzymes mentioned above and also increased the concentration of ascorbic acid to a measurable extent. The role of vitamins in reducing oxidative stress-related effects on spermatogenesis in Cd-treated Swiss mice testes have been reported.

Morphological and genetic analyses of the first record of longrakered trevally, Ulua mentalis (Perciformes: Carangidae) and of the pinjalo snapper, Pinjalo pinjalo (Perciformes: Lutjanidae) in the Odisha coast, Bay of Bengal.

Identification of fish species have so far been carried out mostly by classical morpho-taxonomy. In the present study, however, an attempt has been taken to identify two species of fishes Ulua mentalis and Pinjalo pinjalo of order Perciformes which happens to be the first record in Odisha coast Bay of Bengal, India during the year 2015, using DNA barcoding technique for reconfirmation over conventional morpho-taxonomy. During recent past, study of molecular-taxonomical profile of mitochondrial DNA in general and Cytochrome Oxidase subunit I (COI) gene in particular has gained enormous importance for accurate identification of species. In the present study, the partial COI sequence of Ulua mentalis and Pinjalo pinjalo were generated. Analysis using the COI gene produced phylogenetic trees in concurrence with other multi gene studies and we came across the identical phylogenetic relationship considering Neighbor-Joining and Maximum Likelihood tree. Moreover, these molecular data set further testified in Bayesian framework to reevaluate the exact taxonomic groupings within the family. Surprisingly, Ulua mentalis and Pinjalo pinjalo seems to be closely related to their sister taxa.

Protective action of vitamins on the spermatogenesis in lead-treated Swiss mice.

The protective action of vitamins C and E against lead acetate-induced reduced sperm count and sperm abnormalities in Swiss mice has been studied. Intraperitoneal injection of lead acetate (10mg/kg body weight) in the present study stimulates lipid peroxidation in the testicular tissue, indicated by a significant increase in malondialdehyde content in the experimental mice group. This is associated with an increased generation of noxious reactive oxygen species (ROS). Significantly reduced sperm count associated with increased sperm abnormality percentage in the lead-injected mice group compared to controls substantially proves the ongoing damaging effects of lead-induced ROS on developing germ cells. However, intraperitoneal administration of vitamin C (Vit C) at a concentration equivalent to the human therapeutic dose (10 mg/kg body weight) was able to minimize significantly the testicular malondialdehyde content with a concomitant increase in sperm count and significant decrease in the percentage of abnormal sperm population. Vitamin E (Vit E) (100 mg/kg body weight) treatment of a batch of lead-injected mice had a similar effect as Vit C but with a comparatively lower efficacy. On the other hand, coadministration of both vitamins (Vit C + Vit E) at the above mentioned doses to lead-treated mice led to the most significant decline in malondialdehyde content along with elevated sperm count and reduction in the percentage of abnormal sperm population. The protective action and the synergistic action of both vitamins (C and E) against lead-induced genotoxicity are discussed.

Status of antioxidant defense system in chromium-induced Swiss mice tissues.

Based on epidemiological studies, chromium(VI) compounds are considered as more toxic and carcinogenic than chromium(III) compounds. The deleterious effects of chromium(VI) compounds are diversified affecting almost all the organ systems in a wide variety of animals. The present study, describes the cytotoxic effects of chromium trioxide, a well-known chromium(VI) compound in three tissues (liver, kidney, lungs) of male Swiss mice during post-treatment phase (5th-8th week after treatment). Lipid peroxidation, an index of oxidative stress, was determined as thiobarbituric acid-reactive substances (TBA-Rs) in mice tissues dosed with a single intraperitoneal injection of chromium trioxide (1mg/kg body weight). Tissue specific and statistically significant increases in TBA-Rs was observed in chromium-treated mice groups compared to controls in all the weeks of post-treatment. Endogenous ascorbic acid (vit-C) content of tissues which happens to be one of the stable antioxidants, declined significantly due to chromium-induction. Activity of antioxidative enzymes like superoxide dismutase (SOD), catalase (CT) and peroxidase (PD) was significantly inhibited among chromium-injected mice groups compared to respective controls. Protective role of ascorbic acid and the antioxidative enzymes in chromium-induced cytotoxicity in mice is discussed.

Role of vitamin C on lead acetate induced spermatogenesis in swiss mice.

In the present study significantly increased lipid peroxidation value (LPP) after a single intraperitioneal injection of lead acetate (LA) (100 mg/kg b.w.) indicated enormous generation of Reactive Oxygen Species (ROS). Lead-induced ROS has a direct inhibitory effect on the growth and differentiation of the spermatogonial cells showing a significant decline in sperm count. Chromosomal analysis of the primary spermatocytes at week 4 post-treatment in lead-treated mice revealed significantly higher no of aberrant cells including chromosomal deficiency, autosomal and XY-asynapsis plates compared to untreated control mice, Sperm morphology studies at week 1-4 and at week 8 post-treatment, indicated higher percentage of deformed sperm population compared to vehicle injected groups of mice. Supplementation of vitamin C (Vit C) at a dose of 10 mg/kg body weight to lead-treated mice groups, however, significantly reduced the LPP with a concomitant increase in sperm count, marked decrease in the no of aberrant cells and significant decline in the percentage of morphologically abnormal sperm population. Protective role of Vit C in combating lead-induced oxidative stress in mice testicular cells, has been discussed.

Potential role of vitamins in chromium induced spermatogenesis in Swiss mice.

Chromium (Cr) (VI) compounds are known carcinogens and mutagens. The mechanism of carcinogenicity and mutagenicity caused by chromium(VI) compounds remained unclear for several years. However, in the recent past chromium-induced carcinogenicity and/or mutagenicity was known to happen due to the generation of reactive oxygen species (ROS). In the present context, chromic acid (CrO(3)), a potential Cr(VI) compound could be able to generate reactive oxygen radicals in the testes of Swiss mice as evidenced from significantly higher lipid peroxidation compared to untreated controls. The cytotoxic effects of the compound on the testes are depicted in terms of significantly reduced sperm count level accompanied with increased abnormal sperm population in treated mice. Supplementation of vitamins like Vitamin C and Vitamin E (Vit C and Vit E) to CrO(3) injected mice groups could partially prevent the incidence of abnormal sperm population and increased the sperm count. Of the two vitamins, taken for the study, Vit C happens to be more effective in ameliorating germ cells from degeneration and from mutation to abnormal sperm. Possible antioxidative role of both the vitamins have been studied for significant decrease in lipid peroxidation associated with marked elevation in sperm count level and significant decrease in the percentage of abnormal sperm formation in CrO(3)-treated mice.

Drosophila Sirt2/mammalian SIRT3 deacetylates ATP synthase ß and regulates complex V activity.

Adenosine triphosphate (ATP) synthase ß, the catalytic subunit of mitochondrial complex V, synthesizes ATP. We show that ATP synthase ß is deacetylated by a human nicotinamide adenine dinucleotide (NAD(+))-dependent protein deacetylase, sirtuin 3, and its Drosophila melanogaster homologue, dSirt2. dsirt2 mutant flies displayed increased acetylation of specific Lys residues in ATP synthase ß and decreased complex V activity. Overexpression of dSirt2 increased complex V activity. Substitution of Lys 259 and Lys 480 with Arg in human ATP synthase ß, mimicking deacetylation, increased complex V activity, whereas substitution with Gln, mimicking acetylation, decreased activity. Mass spectrometry and proteomic experiments from wild-type and dsirt2 mitochondria identified the Drosophila mitochondrial acetylome and revealed dSirt2 as an important regulator of mitochondrial energy metabolism. Additionally, we unravel a ceramide-NAD(+)-sirtuin axis wherein increased ceramide, a sphingolipid known to induce stress responses, resulted in depletion of NAD(+) and consequent decrease in sirtuin activity. These results provide insight into sirtuin-mediated regulation of complex V and reveal a novel link between ceramide and Drosophila acetylome.