Toxicity assessment of chronic exposure to common insecticides and bee medications on colony development and drones sperm parameters.

The effect of agrochemicals and beekeeping treatments on drones have not been widely investigated compared to workers or queens. In the present study, we investigated the chronic exposure of chemicals set (deltamethrin, acetamiprid, oxalic acid, fumagillin, and amitraz) on some sperm parameters and on the histomorphology of seminal vesicles. We also assessed the colony development and nosema load before and after the exposure. Thirty native Apis mellifera anatolica honeybee colonies with sister queens equalized with brood and total frame of bees were used for this experiment. Five colonies were used for each group. Deltamethrin, acetamiprid and fumagillin were given as oral chronic exposure at final concentrations of 25.10-6 mg L-1, 0.01 m L-1 and 50 mg L-1 respectively in syrup solution (50/50). Colonies were exposed to oxalic acid by spraying 5 mL per frame space of 3% (w/v) of oxalic acid dihydrate. Finally, the amitraz was applied based on the manufacturer's instructions. The concentrations chosen represented the field-realistic concentrations and those used by beekeepers in the region. Results showed that deltamethrin reduced brood production. In the same group, we found a high increase in nosema load. All treatments decreased sperm count except for fumagillin but this compound increased sperm mortality and increased the percentage of sperm with defected acrosome integrity. The amitraz exhibited a high sperm mortality and high percentage of sperm with defected membrane integrity function. The sperm parameters such as the count, the motility, the acrosome integrity, the membrane function of sperm, and the histomorphology of seminal vesicles of drones exposed to oxalic acid were the most affected. Bee medications commonly used such as oxalic acid and fumagillin should be more investigated and should be considered by beekeepers and particularly queen breeders.

Rapid, practical and safe isolation of adipose derived stem cells.

We compared migration, proliferation, growth curve, confluency and differentiation into adipogenic, osteogenic and chondrogenic cell lineages of mesenchymal stem cells derived from adipose tissue cultured in scratched and nonscratched cell culture flasks. Mesenchymal stem cells were isolated from rat adipose tissue using a nonenzymatic method. We investigated two groups. For the control group, minced adipose tissue was implanted conventionally onto the surface of standard plastic cell culture flasks. For the experimental group, the tissues were cultured in flasks with a scratched surface. We found that scratched flasks promoted cell migration, proliferation and confluency. Our findings suggest that scratched flasks may be used to ensure rapid, practical and safe isolation of adipose tissue-derived stem cells.

Intracerebroventricularly injected nesfatin-1 activates central cyclooxygenase and lipoxygenase pathways.

Nesfatin-1 is a multifunctional neuropeptide having crucial autonomic roles. It is well known that nesfatin-1 collaborates with other central neuromodulatory systems, such as central corticotropin-releasing hormone, melanocortin, oxytocin, and cholinergic systems to show its autonomic effects. Central arachidonic acid cascade plays an important role to provide the homeostasis by exhibiting similar autonomic effects to nesfatin-1. Based on these similarities, the current study was designed to show the effects of intracerebroventricularly (ICV) injected nesfatin-1 on the hypothalamic arachidonic acid (AA) cascade. Immunochemistry and western blot approaches demonstrated that ICV administration of nesfatin-1 provokes an increase in the hypothalamic cyclooxygenase (COX) -1, -2 and lipoxygenase (LOX) protein expression. Moreover, the microdialysis study demonstrated that centrally injected nesfatin-1 increased the posterior hypothalamic extracellular AA products. In conclusion, these findings report that while nesfatin-1 is generating its autonomic effects, it also might be using central prostaglandins and leukotrienes by activating central COX and LOX pathways.

Activation of the central histaminergic system mediates arachidonic-acid-induced cardiovascular effects.

The aim of this study was to explain the involvement of the central histaminergic system in arachidonic acid (AA)-induced cardiovascular effects in normotensive rats using hemodynamic, immunohistochemistry, and microdialysis studies. Intracerebroventricularly (i.c.v.) administered AA (0.25, 0.5, and 1.0 µmol) induced dose- and time-dependent increases in mean arterial pressure and decreased heart rate in conscious normotensive Sprague-Dawley rats. Central injection of AA (0.5 µmol) also increased posterior hypothalamic extracellular histamine levels and produced strong COX-1 but not COX-2 immunoreactivity in the posterior hypothalamus of rats. Moreover, the cardiovascular effects and COX-1 immunoreactivity in the posterior hypothalamus induced by AA (0.5 µmol; i.c.v.) were almost completely blocked by the H2 receptor antagonist ranitidine (50 and 100 nmol; i.c.v.) and partially blocked by the H1 receptor blocker chlorpheniramine (100 nmol; i.c.v.) and the H3-H4 receptor antagonist thioperamide (50 and 100 nmol; i.c.v.). In conclusion, these results indicate that centrally administered AA induces pressor and bradycardic responses in conscious rats. Moreover, we suggest that AA may activate histaminergic neurons and increase extracellular histamine levels, particularly in the posterior hypothalamus. Acting as a neurotransmitter, histamine is potentially involved in AA-induced cardiovascular effects under normotensive conditions.

Centrally administered CDP-choline induced cardiovascular responses are mediated by activation of the central phospholipase-prostaglandin signaling cascade.

The present study was designed to determine the involvement of central prostaglandin synthesis on the pressor and bradycardic effect of cytidine 5'-diphosphocholine (CDP-choline). Intracerebroventricular (i.c.v.) administration of CDP-choline was made and blood pressure and heart rate were recorded in male Sprague Dawley rats throughout this study. Microdialysis and immunohistochemical studies were performed to measure extracellular total prostaglandin concentration and to show cyclooxygenase-1 and -2 (COX-1 and -2) immunoreactivities, respectively, in the posterior hypothalamic area. Moreover, rats were pretreated (i.c.v) with mepacrine [a phospholipase A2 (PLA2) inhibitor], ibuprofen [a nonselective COX inhibitor], neomycine [a phospholipase C (PLC) inhibitor] or furegrelate [a thromboxane A2 (TXA2) synthesis inhibitor] 5 min prior to the injection of CDP-choline to determine the effects of these inhibitors on cardiovascular responses to CDP-choline. Control rats were pretreated (i.c.v) with saline. CDP-choline caused a dose- and time-dependent increase in blood pressure and decrease in heart rate. Immunohistochemical studies showed that CDP-choline increased COX-1 and -2 immunoreactivities in the posterior hypothalamic area. CDP-choline also elevated hypothalamic extracellular total prostaglandin concentration by 62%, as shown in microdialysis studies. Mepacrine or ibuprofen pretreatments almost completely blocked the pressor and bradycardic responses to CDP-choline while neomycine or furegrelate partially attenuated the drug-induced cardiovascular effects. The results suggest that CDP-choline may stimulate prostaglandin synthesis through the activation of PLA2, cyclooxygenases (COX-1 and -2) and prostaglandins and at least TXA2, may mediate the drug׳s cardiovascular effects.

Spermatogonial stem cell sensitivity to capsaicin: an in vitro study.

BACKGROUND: Conflicting reports have been published on the sensitivity of spermatogenesis to capsaicin (CAP), the pungent ingredient of hot chili peppers. Here, the effect of CAP on germ cell survival was investigated by using two testis germ cell lines as a model. As CAP is a potent agonist of the transient receptor potential vanilloid receptor 1 (TRPV1) and no information was available of its expression in germ cells, we also studied the presence of TRPV1 in the cultured cells and in germ cells in situ. METHODS: The rat spermatogonial stem cell lines Gc-5spg and Gc-6spg were used to study the effects of different concentrations of CAP during 24 and 48 h. The response to CAP was first monitored by phase-contrast microscopy. As germ cells appear to undergo apoptosis in the presence of CAP, the activation of caspase 3 was studied using an anti activated caspase 3 antibody or by quantifying the amount of cells with DNA fragmentation using flow cytometry. Immunolocalization was done with an anti-TRPV1 antibody either with the use of confocal microscopy to follow live cell labeling (germ cells) or on Bouin fixed paraffin embedded testicular tissues. The expression of TRPV1 by the cell lines and germ cells was confirmed by Western blots. RESULTS: Initial morphological observations indicated that CAP at concentrations ranging from 150 uM to 250 uM and after 24 and 48 h of exposure, had deleterious apoptotic-like effects on both cell lines: A large population of the CAP treated cell cultures showed signs of DNA fragmentation and caspase 3 activation. Quantification of the effect demonstrated a significant effect of CAP with doses of 150 uM in the Gc-5spg cell line and 200 uM in the Gc-6spg cell line, after 24 h of exposure. The effect was dose and time dependent in both cell lines. TRPV1, the receptor for CAP, was found to be expressed by the spermatogonial stem cells in vitro and also by premeiotic germ cells in situ. CONCLUSION: CAP adversely affects spermatogonial survival in vitro by inducing apoptosis to those cells and TRPV-1, a CAP receptor, may be involved in this effect as this receptor is expressed by mitotic germ cells.

Histological investigations on the effects of feeding a diet containing red hot pepper on the reproductive organs of the chicken.

In the present study the growth and reproductive organ differences in chickens fed a diet containing 1% red hot pepper (10 g/kg diet) from the first day of age were investigated. In birds fed with the experimental diet it was observed that the abdominal fat content decreased. During the experiment the increase in weight gain in the treated group in the first 4 months was reversed in favour of the control group in month 5. Follicular development in the treated group was faster and laying started 11 days before the control group, and the epithelial and muscular development of the oviduct was always greater than that of the control group. The results indicated that red hot pepper consumed in lower concentrations during the development period in the chickens caused faster development of the reproductive system organs. Laying started 11 days earlier in chicks fed with the red hot pepper added diet, an important economic aspect for egg producers, but which may have implications for other animals. A decrease in abdominal fat content and disorders of lipid metabolism are still under investigation.