Nuclear magnetic resonance spectroscopy based metabolomics to identify novel biomarkers of alcohol-dependence

Alcohol misuse is a ravaging public health and social problem. Its harm can affect the drinkers and the whole society. Alcohol-dependence is a phase of alcohol misuse in which the drinker consumes excessive amounts of alcohol and has a continuous urge to consume alcohol. Current methods of alcohol dependence diagnoses are questionnaires and some biomarkers. However, both methods lack specificity and sensitivity. Metabolomics is a scientific field which deals with the identification and the quantification of the metabolites present in the metabolome using spectroscopic techniques such as nuclear magnetic resonance (NMR). Metabolomics helps to indicate the perturbation in the levels of metabolites in cells and tissues due to diseases or ingestion of any substances. NMR is one of the most widely used spectroscopic techniques in metabolomics because of its reproducibility and speed. Some recent metabolomics studies were conducted on alcohol consumption and alcohol misuse in animals and humans. However, few focused on identifying alcohol dependence novel biomarkers. A sensitive and specific technique such as NMR based metabolomics applied to find novel biomarkers in plasma and urine can be useful to diagnose alcohol-dependence

Averrhoa carambola leaves prevent dyslipidemia and oxidative stress in a rat model of poloxamer-407-induced acute hyperlipidemia

Background: The star fruit [Averrhoa carambola L (Oxalidaceae)] is traditionally used in the treatment of many ailments in many countries. It possesses several pharmacological activities, including antioxidant and anti-inflammatory effects. However, it contains the neurotoxic caramboxin and its high content of oxalic acid limits its consumption by individuals with compromised kidney function. This study assessed the anti-hyperlipidemic and antioxidant activities of different fractions of the methanolic extract of A. carambola leaves (MEACL).Methods: The antioxidant activity was investigated using FRAP, and ABTS and DPPH radical-scavenging assays and the inhibitory activity toward pancreatic lipase (PL) and HMG-CoA reductase was assayed in vitro. Acute hyperlipidemia was induced by poloxamer-407 (P-407) in rats and different fractions of MEACL (n-hexane, chloroform, n-butanol, ethyl acetate (EA), water, and chloroform) were orally administered. Cholesterol and triglycerides were determined at 0, 12, 24, and 48 h and LDL-C, vLDL-C, HDL-C, lipid peroxidation (LPO) and antioxidants were assayed after 48 h. The expression of ABCA1, ABCG5, ABCG8, LDL-R, SREBP-1, and SREBP-2 and the activity of HMG-CoA reductase were assayed in the liver of P-407-administered rats treated with the EA fraction.Results: The in vitro data revealed potent radical-scavenging activities of MEACL fractions with the most potent effect showed by the EA fraction that also suppressed the activities of HMG-CoA reductase and PL. In P-407-induced hyperlipidemic rats, all fractions prevented dyslipidemia as shown by the decrease in total cholesterol, triglycerides, LDL-C, vLDL-C and atherogenic index. MEACL and its fractions prevented LPO and boosted GSH, superoxide dismutase, glutathione peroxidase, and catalase in P-407-administered rats. The EA fraction showed more effective anti-hyperlipidemic and antioxidant effects than other fractions and downregulated SREBP-2 while upregulated ABCA1 and LDL-R and ameliorated LPL and HMG-CoA reductase in hyperlipidemic rats.Conclusion: MEACL showed in vitro and in vivo antioxidant activity and the EA fraction significantly ameliorated dyslipidemia in a rat model of P-407-induced acute hyperlipidemia by modulating LPL, PL, HMG-CoA reductase, and cholesterolgenesis-related factors. Therefore, the leaves of A. carambola represent a safe alternative for the star fruit particularly in kidney disease patients, and the EA is the most effective anti-hyperlipidemic and antioxidant fraction

The influence of age on semen parameters, libido and mating behaviour of Siamese Long Tail sheep (Pengaruh umur terhadap ciri-ciri semen, libido dan tabiat mengawan biri-biri Siam Ekor Panjang)

Abstrak Sejumlah 325 sampel semen telah dipungut daripada 36 ekor biri-biri jantan Siam Ekor Panjang (SLT) yang berumur antara 5 hingga 28 bulan. Pemungutan dilakukan selama 8 bulan bagi mengkaji pengaruh umur terhadap ciri-ciri semen, libido dan tabiatnya mengawan. Peratus pergerakan spermatozoa nyata lebih rendah (p <0.05) pada biri-biri jantan yang berumur kurang daripada 10 bulan berbanding dengan yang lebih tua. Telah dicerapkan juga bahawa dengan meningkatnya umur (sehingga 28 bulan) libido dan tabiat mengawan tidak terjejas. Kajian ini menunjukkan bahawa SLT mampu menghasilkan semen pada usia 5 bulan tetapi semen yang benar-benar bermutu hanya dapat dihasilkan apabila ternakan mencapai usia 12 bulan. Abstract A total of 325 semen samples were collected from 36 Siamese Long Tail (SLT) rams ranging in age from 5 to 28 months for 8 months to study the influence of age on semen characteristics, libido and mating behaviour. An increasing trend in semen volume was observed from 5 up to 28 months of age in the SLT rams. The spermatozoa motility percentage was significantly (p <0.05) lower in rams below 10 months of age compared with the older animals. It was also observed that the libido and mating behaviour of the SLT rams were not affected with the advancement of age up to 28 months. This study showed that although the SLT rams could produce semen at the age of 5 months, semen with desirable quality could only be produced when the rams were around 12 months old

Toxicological evaluation of dried Kacangma Herb (Leonurussibiricus)in rats

Kacangma (Leonurus sibiricus L.) is a popular traditional herb that has been consumed for decades by the people of Sarawak as a herbal medicine or culinary ingredient. The toxicity of dried kacangma herb on Sprague Dawley male and female rats was evaluated through 90-day sub-chronic studies. The rats were fed kacangma at the rate of 0.5 (low dose), 5 (medium dose) and 25 (high dose) g/kg body weight. The control groups of rats received only the commercial rat pellet. Minor treatment-related effects were observed for body weights, organ weights and the lipid profile parameters and these did not appear to be of toxicological significance. In the sub-chronic toxicity studies, some indications of renal and liver toxicity were evident in the medium and high dose groups when plasma creatinine and liver enzymes were found to be higher when compared with the control and the low dose groups. The hematology study reveals statistically significant mild anemia in rats from the medium and high dose groups as indicated by decreases in hemoglobin, red blood cell count and packed cell volume (hematocrit value). Administration of kacangma herb at medium and high dose was also found to cause adverse effects in histopathological structure of the liver and kidney of both male and female rats. However, low dose group showed no significant differences compared to the control. Therefore, it is considered safe and less chance of developing toxicity if the herb is consumed at the dose of 0.5 g/kg body weight as observed throughout the 90 days period of sub-chronic study

Lyophilized Hybrid Nanostructured Lipid Carriers to Enhance the Cellular Uptake of Verapamil: Statistical Optimization and In Vitro Evaluation

Abstract Verapamil is a calcium channel blocker and highly effective in the treatment of hypertension, angina pectoris, and other diseases. However, the drug has a low bioavailability of 20 to 35% due to the first pass effect. The main objective of this study was to develop hybrid verapamil-dextran nanostructured lipid carriers (HVD-NLCs) in an attempt to increase verapamil cellular uptake. The formulations were successfully prepared by a high-shear homogenization method and statistically optimized using 24 full factorial design. The HVD-NLCs formulations were freeze-dried using trehalose as a cryoprotectant. The results showed that the optimized formula (VER-9) possessed a particle size (PS), polydispersity index (PDI), and the percentage of entrapment efficiency (%EE) of 192.29 ± 2.98, 0.553 ± 0.075, and 93.26 ± 2.66%, respectively. The incorporation of dextran sulfate in the formulation had prolonged the release of verapamil (~ 85% in 48 h) in the simulated gastric fluid (pH 1.2) and simulated intestinal fluid (pH 6.8). The differential scanning calorimetry analysis showed no chemical interaction between verapamil and the excipients in the formulation. While wide-angle X-ray scattering studies demonstrated the drug in the amorphous form after the incorporation in the NLCs. The transmission electron microscopy and scanning electron microscopy images revealed that the nanoparticles had spherical shape. The cellular uptake study using Caco-2 cell line showed a higher verapamil uptake from HVD-NLCs as compared to verapamil solution and verapamil-dextran complex. The optimized formulation (VER-9) stored in the refrigerated condition (5 °C ± 3 °C) was stable for 6 months. In conclusion, the HVD-NLCs were potential carriers for verapamil as they significantly enhanced the cellular uptake of the drug

Lyophilized hybrid nanostructured lipid carriers to enhance the cellular uptake of verapamil: statistical optimization and in vitro evaluation

Verapamil is a calcium channel blocker and highly effective in the treatment of hypertension, angina pectoris, and other diseases. However, the drug has a low bioavailability of 20 to 35% due to the first pass effect. The main objective of this study was to develop hybrid verapamil-dextran nanostructured lipid carriers (HVD-NLCs) in an attempt to increase verapamil cellular uptake. The formulations were successfully prepared by a high-shear homogenization method and statistically optimized using 24 full factorial design. The HVD-NLCs formulations were freeze-dried using trehalose as a cryoprotectant. The results showed that the optimized formula (VER-9) possessed a particle size (PS), polydispersity index (PDI), and the percentage of entrapment efficiency (%EE) of 192.29 ± 2.98, 0.553 ± 0.075, and 93.26 ± 2.66%, respectively. The incorporation of dextran sulfate in the formulation had prolonged the release of verapamil (~ 85% in 48 h) in the simulated gastric fluid (pH 1.2) and simulated intestinal fluid (pH 6.8). The differential scanning calorimetry analysis showed no chemical interaction between verapamil and the excipients in the formulation. While wide-angle X-ray scattering studies demonstrated the drug in the amorphous form after the incorporation in the NLCs. The transmission electron microscopy and scanning electron microscopy images revealed that the nanoparticles had spherical shape. The cellular uptake study using Caco-2 cell line showed a higher verapamil uptake from HVD-NLCs as compared to verapamil solution and verapamil-dextran complex. The optimized formulation (VER-9) stored in the refrigerated condition (5 °C ± 3 °C) was stable for 6 months. In conclusion, the HVD-NLCs were potential carriers for verapamil as they significantly enhanced the cellular uptake of the drug