Purpose: To investigate the effect of different methods of synchronization on sheep granulosa cell cycle.Materials and Methods: Granulosa cells were aspirated from ovarian follicles and plated in a DMEM medium containing 15% FBS. Upon 70-80% confluency, the medium of the primarycultured as well as the passaged-5 cells were replaced with the medium containing either 0.5% FBS for 24, 48 and 72 h or 0.5 mg mimosine for 24 h. In the last group the cells were cultured in a base medium for further 4 days. In the present investigation, for each culture system, the cells were examined in terms of their cell cycle stage using flow cytometry. Moreover, the cultures were investigated with respect to their apoptotic as well as the proliferating cell contents by using Brdu labeling and TUNEL staining.Results: At primary as well as passaged-5 cultures subjected to serum starvation for 24 h, the frequency of G0/G1, proliferating as well as apoptotic cells were similar to those of control group. At culture with 48 and 72h serum starvation, the percentage of G0/G1 cells tended to increase significantly to 83% and 85% at primary culture and 89% and 90% at passage-5 culture respectively. Moreover, treating the cultures with mimosine caused the G0/G1 cell to increase. The percentages of apoptotic cells in cultures with either serum starvation (for 24 and 48 h) or with mimosine did not increase compared to those of control cultures. According to our results, 72 h after serum starvation, frequency of the apoptotic cells appeared to increase significantly.

Background and Objectives: Mimosa pudica L., belonging to the Fabaceae family, has been used in the Cuban traditional culture by exposure to smoke of aerial parts combustion and in the smokable form of cigarettes, for recreational or leisure purposes and medical applications. The aim of this study was to evaluate the toxicity and physical dependence of M. pudica by the acute inhalation toxicity study and spontaneous withdrawal test in rats exposed to smoke inhalation of the plant. Methods: Mimosine content was determined by a colorimetric method. The acute inhalation toxicity study was performed following the Organization for Economic Cooperation and Development test guidelines 433. The addictive potential of the combustion products of the dried aerial parts, was evaluated through a non-precipitated withdrawal test in female Sprague Dawley rats, in both studies using plant doses of 1000 mg/kg. Results: Quantitative analysis of mimosine content in smoke showed levels of 0. 62±0. 05 µg/mg dried weight. The inhalation toxicity assessment revealed congestion in nasal cavity, focal gliosis in brain and peribronchial pneumonitis. The plant caused signs of physical dependence characterized by the manifestation of hyperactivity (excitability and aggressiveness), piloerection and soft feces, along with decreased body weight and increased rectal temperature during the period of abstinence. Conclusion: Rats exposed to M. pudica showed signs of acute inhalation toxicity similar to smokable substances such as tobacco and marijuana, and evidenced physical dependence comparable to chlordiazepoxide. These findings highlight the potential risks associated with the traditional use of M. pudica in smokable form.

Background: Herbs can be toxic and may be even life-threatening. The mixture of different plants and herbs made by traditional healer and their canvassing on the street attract general people. Here, we report four cases of severe herbal poisoning. Case presentation: In 2008, four young people rushed to DMC Hospital in the early morning with a history of taking herbal medicine (tonic) on that night for gratification. About 3– 4 h after ingestion, they experienced repeated vomiting and abdominal pain. Two patients deteriorated within the hours after admission with restlessness, progressive unconsciousness, and died soon after. The other two patients absconded from the hospital, including the person who prepared the tonic. Screening of the tonic by gas chromatography-mass spectrometry did not reveal toxic components. Discussion: The suspected herbs used for the preparation of that tonic were Santalum album (Chandan wood) which contains Santalol and other etheric oils; Plantago ovata (Ispaghula Husk) containing diverse alkaloids, phenols, etc.; and Mimosa pudica which is the common Mimosa and contains the alkaloid Mimosine. The nature of the tonic and source of the intoxication could not be finally elucidated. Conclusion: The described cases of unknown herbal poisoning in Bangladesh highlight the need for awareness campaigns targeting the population at risk.

Due to rapid aquaculture development, limited supply of fishmeal and its increasing cost and demand, using plant derived feed ingredients such as legumes (lupin and peas), oil seed cakes (soy bean, cottonseed, and rape seed), cereals (corn, rice and wheat), meals of protein rich leaves, concentrates and isolate of non-edible oil seeds (jatropha, castor and neem) as fish feed ingredients is compulsory. However, the major challenge in utilizing the protein rich plant ingredients is the presence of anti-nutritional factors. Anti-nutritional factors are compounds which reduce the nutrient utilization and/or feed intake of aquatic animals and are important in plant origin feed ingredient contents of feed formulation. These factors can cause malnutrition or reduced micronutrients and minerals uptake. The most important anti-nutritional factors include protease inhibitors, phytic acid, saponin, tannin, cyanide, oxalate, gossypol, non-starch polysaccharides, phytoestrogens, mimosine and mycotoxins. There are various traditional and recent technologies, which can be used to reduce the levels of such anti-nutrient factors. Several processing techniques including fermentation, germination, dehulling, autoclaving, soaking etc. are applicable to reduce the anti-nutrient contents of ingredients. However, investigating and appreciation of effective methods and optimizing the conditions of their application is still very demanding. In addition, each aquatic species has tolerable threshold for each of the anti-nutritional substances, which should be considered before choosing the processing method. Other issues including the environmental concerns of the processing method and its economic viability also deserve special attention.