Evaluation of 13-week repeated oral dose toxicity of Areca catechu in F344/N rats.

This study investigated the potential toxicity of the Areca catechu water extract after 13-week repeated oral administration at 0, 166.7, 500, and 1500 mg/kg/day in rats. During the study period, clinical signs, mortality, body weight, food consumption, water consumption, urinalysis, estrous cycle, sperm count and motility, ophthalmoscopy, hematology, serum biochemistry, gross pathology, organ weights, and histopathology were examined. At 1500 mg/kg/day, both sexes exhibited an increase in the incidence of abnormal clinical signs, which included, decreased body weight gain and food consumption, and increased urine bilirubin, ketone bodies, specific gravity, and protein and kidney weight. An increase in liver weight and estrous cycle alterations was observed in females. Serum biochemical and histopathological investigations revealed an increase in the levels of serum aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase, and the incidence of hepatic necrosis in females. At 500 mg/kg/day, an increase in the incidence of abnormal clinical signs including diarrhea and soiled perineal region, was observed in both sexes. No treatmentrelated effects were observed at 166.7 mg/kg/day. Under the present experimental conditions, the target organs were determined to be the liver, kidney, and female reproductive system in rats. The no-observedeffect level was considered to be 166.7 mg/kg/day in rats.

Four-Week Repeated Intravenous Dose Toxicity and Toxicokinetic Study of TS-DP2, a Novel Human Granulocyte Colony Stimulating Factor in Rats.

TS-DP2 is a recombinant human granulocyte colony stimulating factor (rhG-CSF) manufactured by TS Corporation. We conducted a four-week study of TS-DP2 (test article) in repeated intravenous doses in male and female Sprague-Dawley (SD) rats. Lenograstim was used as a reference article and was administered intravenously at a dose of 1000 µg/kg/day. Rats received TS-DP2 intravenously at doses of 250, 500, and 1000 µg/kg/day once daily for 4 weeks, and evaluated following a 2-week recovery period. Edema in the hind limbs and loss of mean body weight and body weight gain were observed in both the highest dose group of TS-DP2 and the lenograstim group in male rats. Fibro-osseous lesions were observed in the lenograstim group in both sexes, and at all groups of TS-DP2 in males, and at doses of TS-DP2 500 µg/kg/day and higher in females. The lesion was considered a toxicological change. Therefore, bone is the primary toxicological target of TS-DP2. The lowest observed adverse effect level (LOAEL) in males was 250 µg/kg/day, and no observed adverse effect level (NOAEL) in females was 250 µg/kg/day in this study. In the toxicokinetic study, the serum concentrations of G-CSF were maintained until 8 hr after administration. The systemic exposures (AUC0-24h and C0) were not markedly different between male and female rats, between the administration periods, or between TS-DP2 and lenograstim. In conclusion, TS-DP2 shows toxicological similarity to lenograstim over 4-weeks of repeated doses in rats.

Target organ identification of jellyfish envenomation using systemic and integrative analyses in anesthetized dogs.

Proper treatment of jellyfish envenomed patients can be successfully achieved only from an understanding of the overall functional changes and alterations in physiological parameters under its envenomation. The majority of previous investigations on jellyfish venoms have covered only a couple of parameters at a time. Unlike most other fragmentary jellyfish studies, we employed an integrative toxicological approach, including hemodynamics, clinical chemistry and hematology analyses, using N. nomurai jellyfish venom (NnV) in dogs. After the baseline measurements for mean arterial pressure (MAP), cardiac output (CO) and heart rate (HR), NnV was intravenously administered to the dogs at doses of 0.1 or 0.3mg/kg body weight. The dogs showed significant decreases in MAP (-27.4±3.7 and -48.1±9.9 mmHg), CO (-1.1±0.1 L/min and -1.0±0.2 L/min), and HR (-4.5±0.3 and -9.9±3.1 beats/min) comparing with the respective baseline controls. The onset of systemic hypotension and bradycardia occurred within 1 min of NnV injection and they lasted for 1-35 min, depending on the NnV doses. Interestingly, serum biochemical analyses of envenomed dogs exhibited dramatic increases of alkaline phosphatase (ALP), creatine phosphokinase (CPK), alanine aminotransferase (ALT) and aspartate aminotransferase (AST), indicating its possible target organs. In conclusion, we have demonstrated simultaneously, for the first time, the multiple organ toxicities (cardiotoxic, myotoxic and hepatotoxic) of a scyphozoan jellyfish venom. Based on these results, an integrative toxinological approach using dogs appears to be effective in predicting jellyfish venom toxicities and designing their therapeutic strategies. We expect this method can be applied to other jellyfish venom research as well.

Enzymatic analysis of Hemiscorpius lepturus scorpion venom using zymography and venom-specific antivenin.

Hemiscorpius lepturus envenomation exhibits various pathological changes in the affected tissues, including skin, blood cells, cardiovascular and central nervous systems. The enzymatic activity and protein component of the venom have not been described previously. In the present study, the electrophoretic profile of H. lepturus venom was determined by SDS-PAGE (12 and 15%), resulting in major protein bands at 3.5-5, 30-35 and 50-60 kDa. The enzymatic activities of the venom was, for the first time, investigated using various zymography techniques, which showed the gelatinolytic, caseinolytic, and hyaluronidase activities mainly at around 50-60 kDa, 30-40 kDa, and 40-50 kDa, respectively. Among these, the proteolytic activities was almost completely disappeared in the presence of a matrix metalloproteinase inhibitor, 1, 10-phenanthroline. Antigen-antibody interactions between the venom and its Iranian antivenin was observed by Western blotting, and it showed several antigenic proteins in the range of 30-160 kDa. This strong antigen-antibody reaction was also demonstrated through an enzyme-linked immunosorbent assay (ELISA). The gelatinase activity of the venom was suppressed by Razi institute polyvalent antivenin, suggesting the inhibitory effect of the antivenin against H. lepturus venom protease activities. Prudently, more extensive clinical studies are necessary for validation of its use in envenomed patients.

Brown alga Ecklonia cava attenuates type 1 diabetes by activating AMPK and Akt signaling pathways.

The antidiabetic therapeutic effect of Ecklonia cava, a brown alga, was investigated using streptozotocin-induced type 1 diabetes mellitus rats and C2C12 myoblasts. The methanol extract of E. cava (ECM), having a strong radical scavenging activity, significantly reduced plasma glucose level and increased insulin concentration in type 1 diabetes mellitus rats. Moreover, the elevation of plasma ALT in diabetic rats was dramatically restored near to normal range by the treatment of ECM, whereas AST level was not meaningfully altered in any group throughout the experiment. The characteristic indications of diabetes, such as polyphagia and polydipsia, were greatly improved by ECM treatment as well. The mechanism of action of ECM appears to be, at least partially, mediated by the activation of both AMP-activated protein kinase/ACC and PI-3 kinase/Akt signal pathways. Taken together, the present results suggest that E. cava has both in vivo and in vitro antidiabetic effects.

Cytotoxicity and hemolytic activity of jellyfish Nemopilema nomurai (Scyphozoa: Rhizostomeae) venom.

The recent bloom of a giant jellyfish Nemopilema nomurai has caused a danger to sea bathers and fishery damages in the waters of China, Korea, and Japan. The present study investigated the cytotoxic and hemolytic activities of crude venom extract of N. nomurai using a number of in vitro assays. The jellyfish venom showed a much higher cytotoxic activity in H9C2 heart myoblast than in C2C12 skeletal myoblast (LC(50)=2 microg/mL vs. 12 microg/mL, respectively), suggesting its possible in vivo selective toxicity on cardiac tissue. This result is consistent with our previous finding that cardiovascular function is a target of the venom. In order to determine the stability of N. nomurai venom, its cytotoxicity was examined under the various temperature and pH conditions. The activity was relatively well retained at low environmental temperature (or=60 degrees C). In pH stability test, the venom has abruptly lost its activity at low pH environment (pH

Mutual synergistic toxicity between environmental toxicants: A study of mercury chloride and 4-nonylphenol.

Mercury chloride (HgCl(2)) and 4-nonylphenol (NP) are widespread environmental and industrial pollutants that are known to have toxic effects as well as endocrine disrupting activities. Although the individual effects of HgCl(2) and NP in liver have been relatively well recognized, little is known about the interaction of NP and HgCl(2) during the induction of their toxicity. In the current study, we investigated the synergism between HgCl(2) and NP using HepG2 cells. Surprisingly, the concurrent treatment of HepG2 with HgCl(2) and NP induced a significant cytotoxicity at concentrations where neither of them have any cytotoxic effect when treated alone. The cytotoxicity of NP is enhanced in the presence of HgCl(2) (a shift from 74.9 to 47.4µM in LC(50)) and vice versa (a shift from 94.9 to 66.3µM in LC(50)). Estrogen receptor antagonists such as ICI 182,780 did not protect HepG2 cells from these cytotoxic insults. Whereas the intracellular level of reduced form glutathione (GSH) was considerably decreased upon the co-treatment with NP and HgCl(2). Furthermore, the synergistic cytotoxicity was significantly inhibited by 20-mM N-acetylcysteine (NAC). These results indicate that the mutual synergistic cytotoxicity of HgCl(2) and NP on HepG2 cell is not associated with estrogen receptor signaling but mediated by reactive oxygen species (ROS) generation. In our real life, we are continuously and often simultaneously exposed to many different kinds of environmental pollutants. The present study suggests a mechanism of potential synergistic adverse effects of these toxic pollutants.