Molecular codes and in vitro generation of hypocretin and melanin concentrating hormone neurons.

Hypocretin/orexin (HCRT) and melanin concentrating hormone (MCH) neuropeptides are exclusively produced by the lateral hypothalamus and play important roles in sleep, metabolism, reward, and motivation. Loss of HCRT (ligands or receptors) causes the sleep disorder narcolepsy with cataplexy in humans and in animal models. How these neuropeptides are produced and involved in diverse functions remain unknown. Here, we developed methods to sort and purify HCRT and MCH neurons from the mouse late embryonic hypothalamus. RNA sequencing revealed key factors of fate determination for HCRT (Peg3, Ahr1, Six6, Nr2f2, and Prrx1) and MCH (Lmx1, Gbx2, and Peg3) neurons. Loss of Peg3 in mice significantly reduces HCRT and MCH cell numbers, while knock-down of a Peg3 ortholog in zebrafish completely abolishes their expression, resulting in a 2-fold increase in sleep amount. We also found that loss of HCRT neurons in Hcrt-ataxin-3 mice results in a specific 50% decrease in another orexigenic neuropeptide, QRFP, that might explain the metabolic syndrome in narcolepsy. The transcriptome results were used to develop protocols for the production of HCRT and MCH neurons from induced pluripotent stem cells and ascorbic acid was found necessary for HCRT and BMP7 for MCH cell differentiation. Our results provide a platform to understand the development and expression of HCRT and MCH and their multiple functions in health and disease

Present state and future perspectives of using pluripotent stem cells in toxicology research

The use of novel drugs and chemicals requires reliable data on their potential toxic effects on humans. Current test systems are mainly based on animals or in vitro–cultured animal-derived cells and do not or not sufficiently mirror the situation in humans. Therefore, in vitro models based on human pluripotent stem cells (hPSCs) have become an attractive alternative. The article summarizes the characteristics of pluripotent stem cells, including embryonic carcinoma and embryonic germ cells, and discusses the potential of pluripotent stem cells for safety pharmacology and toxicology. Special attention is directed to the potential application of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) for the assessment of developmental toxicology as well as cardio- and hepatotoxicology. With respect to embryotoxicology, recent achievements of the embryonic stem cell test (EST) are described and current limitations as well as prospects of embryotoxicity studies using pluripotent stem cells are discussed. Furthermore, recent efforts to establish hPSC-based cell models for testing cardio- and hepatotoxicity are presented. In this context, methods for differentiation and selection of cardiac and hepatic cells from hPSCs are summarized, requirements and implications with respect to the use of these cells in safety pharmacology and toxicology are presented, and future challenges and perspectives of using hPSCs are discussed

Neurobiology of cataplexy.

Cataplexy is the pathognomonic and the most striking symptom of narcolepsy. It has originally been, and still is now, widely considered as an abnormal manifestation of rapid eye movement (REM) sleep during wakefulness due to the typical muscle atonia. The neurocircuits of cataplexy, originally confined to the brainstem as those of REM sleep atonia, now include the hypothalamus, dorsal raphe (DR), amygdala and frontal cortex, and its neurochemistry originally focused on catecholamines and acetylcholine now extend to hypocretin (HCRT) and other neuromodulators. Here, we review the neuroanatomy and neurochemistry of cataplexy and propose that cataplexy is a distinct brain state that, despite similarities with REM sleep, involves cataplexy-specific features

The effects of Matricaria chamomilla L. hydroalcoholic extract on atherosclerotic plaques, antioxidant activity, lipid profile and inflammatory indicators in rats

Introduction: Treatment of cardiovascular risk factors seems to be necessary and involves a number of changes in drug treatment and lifestyle. This study aimed to evaluate the effects of Matricaria chamomilla L. hydroalcoholic extract on antioxidant activity, atherosclerotic plaques, lipid profile and inflammatory indicators in rats. Methods: Thirty male Wistar rats were divided into five experimental groups consisting of group 1 (Sham; normal dietary), group 2 (control; high cholesterol diet (2)), group 3 (high cholesterol diet plus 55 mg/kg of chamomile hydroalcoholic extract), group 4 (high cholesterol diet plus 110 mg/kg of chamomile hydroalcoholic extract), and group 5 (high cholesterol diet plus 10 mg/kg of lovastatin). At the beginning and end of the study, blood samples of all the animals were taken for determination of antioxidant activity and the level of biochemical parameters. The hearts and aorta were also isolated for ontological tests. Results: No symptom of plaque formation was observed in experimental groups 3, 4 and 5 that received the high cholesterol diet. High cholesterol diet (2) resulted in a significant increase in serum cholesterol level, TG and LDL-c levels in groups 2 and 3 as compared to group 1 (P<0.001). No significant difference was observed in serum cholesterol, TG and LDL-c levels in experimental groups 4 and 5, compared to experimental group 1. In group 4, serum HDL-c concentration did not show significant changes as compared to group 1. In groups 4 and 5, no significant change was observed in inflammatory factors as compared to group 1. The levels of superoxide dismutase in red blood cells and malon-dialdehyde in plasma of groups 3 and 5 showed no significant change when compared with group 1. Conclusion. Chamomile led to the management and correction of changes in risk factors of cardiovascular diseases

INVESTIGATING THE METHADONE EFFECTS ON AST, ALT AND ALP ENZYMES OF SERUM OF MALE WISTAR RATS

Narcotics abuse and addiction have been turned to a major problem in the communities. Addiction to any type of narcotics is known to be followed by side effects both physically and psychologically. One way to control addiction is the use of another maintenance medication. The current research paper aims at investigating the effect of one such a drug as methadone on the serum levels of Alkaline phosphatase, Alanine aminotransferase, Aspartate aminotransferase enzymes. A total number of 28 male adult Wistar rats were completely haphazardly assigned to four group (n=7): a control group that received an ordinary daily dietary regime and Experimental groups one, two and three that were administered through oral gavage with 5mg/ml, 20mg/ml and 40mg/ml of methadone syrup, respectively, on a daily basis besides the ordinary dietary regime for eight weeks. Blood samples were collected from the animals at the beginning and at the end of the study for the determination of serum levels of alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase enzymes. In the end, the data were analyzed via taking advantage of one-way variance analysis (one-way ANOVA) and TUKEY Test. The serum levels of all three ALT, AST and ALP enzymes were significantly increased in the groups that had been given different doses of methadone (P < 0.001). According to the results obtained herein and the effects that methadone has on the elevation of liver enzymes levels and also due to the fact that this maintenance medication is being increasingly administered on a daily basis for the treatment of addiction to various narcotics, it seems that there is a need for devising better strategies and better planning for the amount and method of methadone use as well as the way this medication has to be distributed within the society

INVESTIGATION THE ANTI-HYPERLIPIDEMIC EFFECTS OF SAFFRON PETALS' OF HYDROALCOHOLIC EXTRACT IN RATS

The use of medicinal herbs has been customary in traditional medicine since very long ago and according to the intolerances exhibited by some of the patients for chemical drugs due to the emergence of side effects, the medicinal herbs can be utilized as alternative treatments through adhering to evidence-based medical principles. The current research paper investigates the effects of saffron petal hydroalcoholic extract on the lipid profiles. Thirty Wistar rats were assigned to six groups: a control group and a sham group that received ordinary dietary regime and high cholesterol (2) dietary regime, respectively, and experimental groups one to four that, besides the high cholesterol dietary regime, were also, correspondingly, treated by 50, 100 and 200 mg/kg of their body weights saffron petal hydroalcoholic extract per day and 10 mg/kg of the body weight lovastatin per day. Blood samples were collected of the animals after eight weeks to determine their serum lipid profiles. SPSS (version16) was employed to perform the analysis. The serum levels of cholesterol, triglyceride and LDL-C were found significantly increased in sham group and experimental group three (high cholesterol dietary regime plus 200 mg/kg of the extract) in contrast to the control group (P<0.001). The significance level was found lower in experimental group 1 (high cholesterol regime plus 50 mg/kg of the extract) (P<0.05, P<0.001 and P<0.05, respectively). The serum levels of these three factors and HDL-c were not found significantly different in experimental groups two and four in comparison to the control group. The results are indicative of the favorable effects of saffron petal hydroalcoholic extract, especially in lower dosages, on the regulation and correction of the serum lipid profile

The effect of hydroalcoholic extract of Teucrium polium L. on the inflammatory markers and lipid profile in hypercholesterolemic rats

Background: Cardiovascular diseases are among the most common causes of mortality worldwide. Therefore, it is necessary to control the risk factors of these patients. Since the level of inflammatory markers and lipid profiles has increased in cardiovascular diseases and due to the increasing role of plants in the treatment of diseases, the current study aimed to investigate the effect of hydroalcoholic extract of Teucrium polium on inflammatory markers and lipid profile in hypercholesterolemic rats. Materials and methods: A total of 24 adult male Wistar rats were randomly divided into four groups of six each and treated with oral administration for 8 weeks. The control group received normal diet, the sham group received high-cholesterol diet and experimental groups 1 and 2 received high-cholesterol diet in the 8 weeks and doses of 85 and 170 mg/kg, respectively, of the T. polium hydroalcoholic extract (TPHAE) in the second 4 weeks. At the beginning and the end of the study, rats were examined for biochemical parameters. The mean level of variables for each group was presented as mean +/- standard error of mean. Results: The results of this study showed that, after administration of TPHAE, there was a significant decrease in the mean of inflammatory markers in all groups compared to sham group (P<0.001). Also, administration of the extract significantly reduced the serum levels of triglyceride, cholesterol and LDL-cholesterol and significantly increased the serum HDL-cholesterol levels. In addition, the 170 mg/kg dose of TPHAE was the most effective in reducing serum levels of inflammatory and lipid markers. Conclusion: Treatment with TPHAE caused dose-dependent decrease in serum levels of inflammatory markers and lipid profile in hypercholesterolemic rats. Therefore, it can be applied as a natural product for the management of cardiovascular diseases

ANTIHYPERLIPIDEMIC EFFECTS OF HYDROALCOHOLIC LEAF EXTRACT OF TEUCRIUM POLIUM IN HYPERCHOLESTEROLEMIC RATS

Hyperlipidemia is one of the most important factors contributing to the cardiovascular diseases. There are various chemical drugs for regulating the lipid profile level. According to the side effects proved for the chemical drugs, we, in the current research paper, investigate the effect of wall germander (Teucrium polium) hydroalcoholic extract effect on the lipid profile in the hypercholesterolemic rats. Wistar male rats, ranging in weight from 150 g to 180 g, were assigned to four groups (n=5): control group (ordinary diet), sham group (high, 2, cholesterol dietary regime), experimental group one (high, 2, cholesterol dietary regime plus 0.85mg/ml Teucrium polium hydroalcoholic extract) and experimental group two (high, 2, cholesterol dietary regime plus 1.7mg/ml Teucrium polium hydroalcoholic extract). The treatment was run in an eight-week period with the termination of which blood samples were collected from the animals. To determine the lipid profile, commercial diagnostic kits were applied. The results were analyzed by the use of SPSS software, ver. 16. From the week six on, the weights of the rats from the experimental group two (high, 2, cholesterol diet plus 1.7 mg/ml Teucrium polium hydroalcoholic extract) showed a significant decrease (P<0.05) even in comparison to the control group (ordinary diet). The serum level of triglycerides, cholesterol and LDL-c was found significantly decreased in the experimental group two (P<0.001) as compared to Sham group but it did not any significant change in contrast to the control group. In experimental group one (high, 2, cholesterol dietary regime plus 0.85 mg/ml Teucrium polium hydroalcoholic extract), only the cholesterol and TG serum levels were found significantly decreased in respect to the sham group (P<0.05 and P<0.001, respectively). Regarding the lipoprotein amount, a significant increase was documented in the experimental group two, featuring high density cholesterol (HDL-c), in contrast to the sham group (P<0.001). The present study's findings indicated that an appropriate dosage of T. Polium can cause the regulation of lipid profile as well as the weights of the hypercholesterolemic rats. According to the side effects of the chemical drugs similar in their functions to the effect of T. polium, it can be suggested as an antihyperlipidemic drug

The association of diabetes mellitus and gender with risk of bone fractures: A systematic review and meta-analysis

Bone fracture shows a growing trend. This increase is a result of several factors occurs. In this study, a meta-analysis was performed to examine the relationship of fracture risk with Type 2 diabetes mellitus and gender. For this purpose, we searched the credible databases including PubMed, ISI, and Scopus for the related epidemiologic studies that were published up to May 2016. The data provided in these articles was extracted and then analyzed by a random-effects model. Funnel plot was used to check for publication bias. Statistical analyses were carried out by R version 3.2.1 and STATA (version 11.1). After filtering irrelevant articles, 30 studies were selected for meta-analysis. The analyses showed that type 2 diabetes mellitus is positively associated with overall fracture risk (summary RR. 1.05, 95 CI: 1.04, 1.06). The incidence of fractures was higher in men than in women (Phet=0.043). The interpretation of the meta-regression showed that no significant relationship exists between the gender risk factor in diabetic patients and years of study (P=0.280). The results highlight the importance of adopting suitable strategies for prevention of bone fracture in T2DM patients

Hypocretin (orexin) biology and the pathophysiology of narcolepsy with cataplexy.

The discovery of hypocretins (orexins) and their causal implication in narcolepsy is the most important advance in sleep research and sleep medicine since the discovery of rapid eye movement sleep. Narcolepsy with cataplexy is caused by hypocretin deficiency owing to destruction of most of the hypocretin-producing neurons in the hypothalamus. Ablation of hypocretin or hypocretin receptors also leads to narcolepsy phenotypes in animal models. Although the exact mechanism of hypocretin deficiency is unknown, evidence from the past 20 years strongly favours an immune-mediated or autoimmune attack, targeting specifically hypocretin neurons in genetically predisposed individuals. These neurons form an extensive network of projections throughout the brain and show activity linked to motivational behaviours. The hypothesis that a targeted immune-mediated or autoimmune attack causes the specific degeneration of hypocretin neurons arose mainly through the discovery of genetic associations, first with the HLA-DQB1*06:02 allele and then with the T-cell receptor α locus. Guided by these genetic findings and now awaiting experimental testing are models of the possible immune mechanisms by which a specific and localised brain cell population could become targeted by T-cell subsets. Great hopes for the identification of new targets for therapeutic intervention in narcolepsy also reside in the development of patient-derived induced pluripotent stem cell systems