Secalonic acid D; A cytotoxic constituent from marine lichen-derived fungusGliocladium sp. T31

Secalonic acid D (SAD) was isolated as the major secondary metabolite of the marine lichen-derived fungus Gliocladium sp. T31. Its structure was established on the basic of physico-chemical and spectroscopic data. This is the first report on the isolation of SAD from this fungus, as well as its inhibitory effect on K562 cell cycle and its cytotoxicity against several tumor cell lines in vitro.     

Adenohypophysis and growth in tadpoles of Rana temporaria.

To investigate a possible estrogen-like compound in ovary of shrimp, Parapenaeus fissurus was extracted with acetone, and after acid hydrolysis, purified by chromatography. The sample extract was assayed by biological, chemical, and immunological methods. With bioassay the shrimp ovarian extract showed estrogen-like activity of 3.26 ng/g of tissue. The shrimp extract displayed maximum uv absorption at 280 nm and showed absorption at 518 nm after reacting with Kober reagent, properties similar to those of estrogens. The ovarian extract was analyzed by gas chromatography. A peak was identified that corresponded to estrone. The extract was further assayed by an immunological method. It was found by such criteria that estrone and estradiol-17β exist in shrimp ovary, and the former was the major compound. According to these data, it is concluded that an estrogen-like compound exists in ovary of shrimp Parapenaeus fissurus.     

Positive and Negative Brain Zones in the Snail

Fine wire electrodes were surgically implanted in two regions of the brain of the snail Helix aspersa. To receive electrical stimulation of the brain, a tethered snail was required to displace the end of a rod. Self-stimulation delivered to the parietal ganglion resulted in non-repetition of the operant response, whereas self-stimulation delivered to the mesocerebrum resulted in an increase in response frequency. The reinforcing effect of local extracellular stimulation of two brain zones was investigated in a semi-intact preparation of a closely related species with an identical cellular map ( H.lucorum ). It was found that mesocerebral stimulation increased the frequency of the reinforced spontaneous movement, but decreased the frequency of the same movement if its absence was reinforced. These results allow us to attribute positive reinforcing effects to this brain area, which is involved in the control of sexual behaviour. Different results were obtained by contingent stimulation of the rostral part of the parietal ganglia, where giant cells controlling avoidance behaviour are located. Stimulation of this zone resulted in a decrease in the frequency of the ongoing spontaneous movements. These findings make possible intracellular investigations of the mechanisms of positive and negative reinforcement.     

Micronucleus test in mussels Perna perna fed with the toxic dinoflagellate Prorocentrum lima

Micronucleus (MN) and other nuclear abnormalities have been measured in the hemocytes of mussels Perna perna to verify whether feeding mussels with different concentrations of Prorocentrum lima results in accumulation of levels of okadaic acid (OA) capable of inducing genotoxic effects at the chromosome level, as evidenced by micronuclei and nuclear abnormalities. Four groups of 12 mussels housed individually in aquaria containing filtered seawater were fed with different concentrations of P. lima. Another group collected directly from the production area served as outdoor control. A significantly higher frequency of MN and nuclear lesions was observed in hemocytes from the groups fed P. lima.     

In-vitro antimicrobial activity of marine actinobacteria against multidrug resistance Staphylococcus aureus

Objective

To investigate the antibacterial activity of marine actinobacteria against multidrug resistance Staphylococcus aureus (MDRSA).

Methods

Fifty one actinobacterial strains were isolated from salt pans soil, costal area in Kothapattanam, Ongole, Andhra Pradesh. Primary screening was done using cross-streak method against MDRSA. The bioactive compounds are extracted from efficient actinobacteria using solvent extraction. The antimicrobial activity of crude and solvent extracts was performed using Kirby-Bauer method. MIC for ethyl acetate extract was determined by modified agar well diffusion method. The potent actinobacteria are identified using Nonomura key, Shirling and Gottlieb 1966 with Bergey''s manual of determinative bacteriology.

Results

Among the fifty one isolates screened for antibacterial activity, SRB25 were found efficient against MDRSA. The ethyl acetate extracts showed high inhibition against test organism. MIC test was performed with the ethyl acetate extract against MDRSA and found to be 1 000 µg/mL. The isolated actinobacteria are identified as Streptomyces sp with the help of Nonomura key.

Conclusions

The current investigation reveals that the marine actinobacteria from salt pan environment can be able to produce new drug molecules against drug resistant microorganisms.     

Heteronemin, a spongean sesterterpene, inhibits TNFα-induced NF-κB activation through proteasome inhibition and induces apoptotic cell death

In this study, we investigated the biological effects of heteronemin, a marine sesterterpene isolated from the sponge Hyrtios sp. on chronic myelogenous leukemia cells. To gain further insight into the molecular mechanisms triggered by this compound, we initially performed DNA microarray profiling and determined which genes respond to heteronemin stimulation in TNFα-treated cells and which genes display an interaction effect between heteronemin and TNFα. Within the differentially regulated genes, we found that heteronemin was affecting cellular processes including cell cycle, apoptosis, mitogen-activated protein kinases (MAPKs) pathway and the nuclear factor κB (NF-κB) signaling cascade.We confirmed in silico experiments regarding NF-κB inhibition by reporter gene analysis, electrophoretic mobility shift analysis and I-κB degradation. In order to assess the underlying molecular mechanisms, we determined that heteronemin inhibits both trypsin and chymotrypsin-like proteasome activity at an IC₅₀ of 0.4 μM. Concomitant to the inhibition of the NF-κB pathway, we also observed a reduction in cellular viability. Heteronemin induces apoptosis as shown by annexin V-FITC/propidium iodide-staining, nuclear morphology analysis, pro-caspase-3, -8 and -9 and poly(ADP-ribose) polymerase (PARP) cleavage as well as truncation of Bid. Altogether, results show that this compound has potential as anti-inflammatory and anti-cancer agent.     

Clinico-toxinological characterization of the acute effects of the venom of the marine snail, Conus loroisii

The venom of the marine snail, Conus loroisii, was studied to assess its risk and lethal factors in regard of human welfare. The lethality of the crude venom (LD50-5.0 mg/kg via i.p.) in mice was associated with reduced motor activity, asphyxiation, followed by respiratory failure. The effects on vital tissues revealed vascular congestion and inflammatory cell infiltration around the portal triad of the liver, spongiosis of the brain, hemorrhages/congested blood vessels in lung and endothelial cells of the renal tubule. Repeated measures of hematological profiles indicated that the venom significantly reduced erythrocytes (P<0.001, GLM repeated measures), followed associated with depletion of hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin and platelet count. Serum enzymes such as, glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase, lactate dehydrogenase and alkaline and acid phosphatases were altered significantly (P<0.05, Friedman test), which in turn confirmed the damage of vital organ tissues. Dual effect of the venom on the activity of mouse brain acetylcholinesterase stand for concentration specific, whereas maximal inhibition (60.41%, P<0.05, Wilcoxon signed rank test) in erythrocyte acetylcholinesterase did not show the dual activity observed in brain. The Ciphergen ProteinChip analysis of the envenomed serum further revealed that the venom causes changes in definite molecules involved in inflammatory process and ionic transport. In all, the venom of C. loroisii is potentially lethal to mammals, through its rapid action on the central and peripheral nervous systems by blocking neurotransmission with selective interference of ionic channels/receptors.     

Sex steroid concentrations and localization of steroidogenic enzyme expression in free-ranging female northern fur seals (Callorhinus ursinus)

Recent precipitous population declines in northern fur seal (Callorhinus ursinus) and other Alaskan pinniped populations are due in part to reduced fecundity and have emphasized deficits in basic reproductive knowledge of these species. Following estrus and mating, fertilized female pinnipeds experience an obligatory delayed implantation lasting several months and non-pregnant (pseudopregnant) females are indistinguishable by sex steroid levels during this time. The current study examined circulating steroid concentrations and ovarian expression of key steroid-synthesizing enzymes in northern fur seals to identify changes associated with embryonic implantation, data necessary for estimating early pregnancy rates of the population. Blood samples were collected from 84 female fur seals captured on an Alaskan rookery from October 15 to November 30, a period spanning the end of the delay and subsequent uterine implantation of the fertilized blastocyst in this species. Concentrations of progesterone, estradiol, and estrone measured in blood collected during boreal fall were contrasted with samples collected during late summer from post-ovulatory females and males. Serum sex steroids were also related to ovarian expression of key enzymes responsible for androgen (17alpha-hydroxylase/17,20-lyase, cytochrome b5, and 3beta-hydroxysteroid dehydrogenase) and estrogen (aromatase cytochrome P450) synthesis. Enzymes necessary for androgen synthesis were highly expressed in ovaries, and accordingly, endocrine profiles were expanded to include DHEA, androstenediol, androstenedione, and testosterone. Estradiol concentrations were universally low and free and conjugated estrone were the primary circulating estrogens in fur seal sera. No implantation-associated peak was identified for estrogens or progesterone and mean values of progesterone and estrone were actually greater in female serum samples collected during summer than fall. However, there was a significant positive relationship between fall sampling date and testosterone concentration. Additionally, DHEA concentrations in females were lower in samples collected during the implantation period than during the summer embryonic delay. These data suggest androgens may play a substantial regulatory role in the embryonic delay of northern fur seals.