Assessment of vitamin B12 levels and cardiovascular risk factors in metformin- and non-metformin-treated type 2 diabetic patients.

Oxidative stress enhances cardiovascular risk. Metformin decreases intestinal absorption of vitamin B12. Our objective was the evaluation of type 2 diabetics focusing on differences due to their treatment. A prospective study on 224 type 2 diabetics was realized between 2015-2018 in Targu Mures, Romania, divided into 2 subgroups (metformin vs. other therapy - 2nd/3rd generation sulfonylureas, insulin, dietary regimen -, followed for at least one year) and non-diabetic controls (n=25) for oxidative stress level comparison. Serum homocysteine (HC), vitamin B12 were determined by chemiluminescence (Immulite One). Lipid peroxidation was assessed by serum malondialdehyde (MDA) measurement (HPLC). Biochemical tests, minerals, cystatin C, high-sensitivity C reactive protein (hs-CRP) were measured on Konelab20Xti, glycated hemoglobin on Nycocard Reader. GraphPad InStat-3 was used for statistics. Negative correlation occured between serum vitamin B12 and HC, this vitamin's level was significantly lower and serum zinc was significantly higher in patients on metformin. Hyperhomocysteinemia was present in 87% of the subjects, 46% had zinc deficiency and 41% elevated hs-CRP. Serum cystatin C showed positive correlation with creatinine. Serum MDA was significantly higher in diabetics compared to control patients. Elevated hs-CRP and homocysteine represent raised cardiovascular risk. Intense oxidative stress, vitamin, mineral deficiencies are frequent in diabetic subjects.

Profiling the Concentration of Reduced and Oxidized Glutathione in Rat Brain Using HPLC/DAD Chromatographic System.

This study aimed to develop a HPLC/DAD method in order to determine and quantify the reduced glutathione (GSH) and oxidized glutathione (GSSG) levels in rat brain. Due to the presence of the thiol group (-SH), GSH can interact with the Ellman's reagent (DTNB), with which it forms a reaction product through which the level of GSH can be quantified, using the DAD detection system. Chromatographic separation was achieved after a derivatization process by using a mobile phase acetonitrile (A) and phosphate buffer (20 mM, pH = 2.5) (B). The compounds of interest were detected at 330 nm using a chromatographic C8 column. The method of determination met the validation criteria, specified by the regulatory bodies. The applicability of the method was demonstrated in a chronic toxicology study of central nervous system (CNS), following different treatment regimens with haloperidol.

A Simple HPLC/DAD Method Validation for the Quantification of Malondialdehyde in Rodent's Brain.

In the present study, a HPLC/DAD method was set up to allow for the determination and quantification of malondialdehyde (MDA) in the brain of rodents (rats). Chromatographic separation was achieved on Supelcosil LC-18 (3 µm) SUPELCO Column 3.3 cm × 4.6 mm and Supelco Column Saver 0.5 µm filter by using a mobile phase acetonitrile (A) and phosphate buffer (20 mM, pH = 6) (B). Isocratic elution was 14% for (A) and 86% for (B). The injection volume (loop mode) was 100 µL with an analysis time of 1.5 min. Flow rate was set at 1 mL/min. The eluted compound was detected at 532 nm by a DAD detector by keeping the column oven at room temperature. The results indicated that the method has good linearity in the range of 0.2-20 µg/g. Both intra- and inter-day precision, expressed as RSD, were ≤15% and the accuracies ranged between ±15%. The lower limit of quantification (LLOQ), stability, and robustness were evaluated and satisfied the validation criteria. The method was successfully applied in a study of chronic toxicology following different treatment regimens with haloperidol and metformin.

Oxidative stress and peripartum outcomes (Review).

Pregnancy, labor and childbirth are accompanied by excessive oxidative aggression. The excessive formation of free radicals [reactive oxygen species (ROS), reactive nitrogen species (RNS), chlorine reactive species (CRS)] causes cellular oxidative damage, which can be scavenged by enzymatic or non-enzymatic antioxidants in normal healthy pregnancy, physiological labor and delivery without any complications. An imbalance between the pro-oxidant and antioxidant factors may lead to oxidative stress, which contributes to the development of many diseases. This oxidative aggression can be a precursor for pathologies in the pregnant woman including eclampsia, miscarriage, preterm labor, and intrauterine growth retardation; in the offspring it may lead to bronchopulmonary dysplasia/chronic lung disease, necrotizing enterocolitis, retinopathy of prematurity, and periventricular leukomalacia. This review summarizes the studies conducted to identify the mechanisms of oxidative stress and the effect of cell membrane oxidation, the mechanisms that are behind oxidative stress-related diseases, and also those studies which have demonstrated the effect of antioxidants in preventing diseases or diminishing the effects of oxidative stress in the body, in obstetrics and neonatology.

Liquid chromatographic method for the simultaneous determination of achiral and chiral impurities of dapoxetine in approved and counterfeit products.

A reversed-phase high performance liquid chromatographic method was developed and validated for the simultaneous determination of the related substances of S-dapoxetine, including R-dapoxetine, (3S)-3-(dimethylamino-3-phenyl-1-propanol), S-3-amino-3-phenyl-1-propanol, 1-naphtol, 4-phenyl-2H,3H,4H-naphtho[1,2-b]pyran and 1-(2E)-Cinnamyloxynaphthalene. During the screening experiments seven different polysaccharide-type chiral stationary phases (amylose-based Lux-Amylose-1, Lux-i-Amylose-1 and Lux-Amylose-2, as well as cellulose-based Lux-Cellulose-1, Lux-Cellulose-2, Lux-Cellulose-3 and Lux-Cellulose-4) were tested in polar organic mode using a mobile phase consisting of 0.1% diethylamine in methanol, ethanol, 2-propanol and acetonitrile with 0.5 mL min-1 flow rate at 20 °C. Best results were obtained on Lux Cellulose-3 column with the ethanol-based mobile phase. To increase the retention factor of two, early-eluting impurities, water was added to the mobile phase. In order to counterbalance the increased total analysis time, higher column temperature (40 °C) and gradient elution, combined with flow-programming` was applied. Using the optimized conditions baseline separations were achieved for all compounds within 30 min. The method was validated according to the International Council on Harmonization guideline Q2(R1) and applied to the analysis of an approved, tablet formulation and dapoxetine-containing products sold on the internet. As expected, in the case of the pharmacy-acquired product, all of the monitored impurities were below 0.1%. However, interesting results were obtained when internet-acquired samples were analyzed. These tablets contained racemic dapoxetine and/or high concentration of R-dapoxetine impurity. Based on this work polysaccharide-based chiral stationary phases can be successfully applied for the simultaneous determination of achiral and chiral impurities in reversed-phase mode applying gradient elution and flow-rate programs. The study further underlines the importance of not only achiral, but also enantiomeric quality control, whenever counterfeiting of a single enantiomeric agent is suspected.

Characterization of the Site-Specific Acid-Base Equilibria of 3-Nitrotyrosine.

The complete macro- and microequilibrium analyses of 3-nitrotyrosine, a biomarker of oxidative stress damage, are presented for the first time. The protonation macroconstants were determined by 1 H-NMR-pH titration, while microconstants were elucidated by a combination of deductive and NMR methods, in which properties of the methyl ester derivative as an auxiliary compound were also studied. Combination of the NMR-pH characterization of the title and auxiliary compounds and the pair-interactivity parameters of 3-iodotyrosine provided the sufficient system to evaluate all the microconstants. NMR-pH profiles, macroscopic and microscopic protonation schemes, and species-specific distribution diagrams are included. The phenolate basicity of 3-nitrotyrosine is 500 times below that of tyrosine, and it is even lower than that of 3-iodotyrosine. This phenomenon can be explained by the stronger electron withdrawing and the negative mesomeric effect of the nitro group. Based on our results, 89 % of the phenolic OH groups are deprotonated in 3-NT molecules at the pH of the blood plasma.

Extractability of polyphenols from black currant, red currant and gooseberry and their antioxidant activity.

In this study, we analyzed extracts of Ribes (black currant, red currant and gooseberry) fruits obtained with methanol, methanol 50% and water. For each extract total polyphenol content, total flavonoid content and total anthocyanin content was assessed. The antioxidant activity of extracts was evaluated by 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging capacity and by the photo-chemiluminescence (PCL) method. Identification and quantification of individual phenolic compounds was performed by means of high performance liquid chromatograph coupled with diode array detector (HPLC-DAD) analyses. From each fruit, best extraction of polyphenols was obtained with methanol 50%. In case of red currants and gooseberry there was no significant difference in flavonoids and anthocyanins extraction rate by the different extraction solvents. For black currants the methanol and methanol 50% extract presented the highest antioxidant activity. For red currants extracts with methanol 50% showed stronger antioxidant activity (IC50 = 5.71 mg/ml for DPPH, IC50 = 1.17 mg/ml for ABTS) than those with methanol or water. In case of gooseberry by the DPPH test the water extract proved to be the most active (IC50 = 5.9 mg/ml). In the PCL test black currants methanol 50% extract was over 6 times more powerful as the ones from red currants. In case of gooseberries, water extract presented the highest antioxidant activity (41.84 µmol AAE/g). In black currant cyanidin-3-glucoside was the major compound. Quercetin 3-O-glucoside was identified in each sample. From cinnamic acid derivatives neochlorogenic acid was present in black currants in the highest amount (356.33 µg/g).

Vaccinium Extracts as Modulators in Experimental Type 1 Diabetes.

Antihyperglycemic effects of four extracts obtained from leaves and fruits of Vaccinium myrtillus and Vaccinium corymbosum were assessed in diabetic rats. In addition, the effects of extracts on diabetic-related complications such as the development of diabetic cataract and oxidative stress were evaluated. Type 1 diabetes was induced with a single dose of streptozotocin in Wistar rats. The rats were randomly divided into seven equal groups: NC-normal control, DC-diabetic control, PC-positive control treated with metformin, VML-received V. myrtillus leaf extract, VMLF-received VML and fruit extract, VCL-received V. corymbosum leaf extract, and VCLF-received VCL and fruit extract. Body weight and glucose levels were monitored every second week. After 8 weeks of treatment, serum glucose, insulin, and malondialdehyde were measured. Lenses were removed after sacrifice and eight lenses from each group were randomly selected for evaluation of cataract development. A decrease in body weight was observed in all diabetic groups in the first weeks. In the VML group, no significant decrease was observed. Glucose levels during the experiment were high in DC, PC, and VCL groups, with no improvement during the 8 weeks. In VML, VMLF, and VCLF groups, a decrease in blood glucose levels was observed. Similar results regarding serum insulin and glucose levels at the end of the experiment were observed within groups. V. myrtillus extracts prevented the development of cataract compared with the DC group (P < .05).

Smoking habits of pharmacy students attending the University of Medicine and Pharmacy in Targu Mures.

Smoking is common among health professional students. The aim of this study was to assess the smoking habits of the pharmacy students attending the University of Medicine and Pharmacy in Târgu Mureș (UMPh TM), Romania. Material and methods: Smoking habits and attitudes toward smoking among 414 pharmacy students attending UMPh TM (86% female) were evaluated using a self-completed questionnaire. Results: The rate of smoking increases during the time students attend the university (24.1% to 33.3% from 1st to 5th year) and males are significantly are more likely to smoke than females (41.4% vs. 27.3%, p=0.042). 36.9% of the smoking pharmacy students are tobacco-dependent, and 40.4% of smokers started daily smoking at the age of 16-19. We found significant differences between smoker and non-smoker pharmacy students regarding their attitudes toward smoking and tobacco control policies, with non-smokers being more supportive of smoke-free policies. Conclusions: Prevention programs and education have a very important role in decreasing the percentage of smokers and support for smokefree policies, but it is critical to begin such programs early in their university training.

Thinking about the past, present and future in adolescents growing up in Children's Homes.

The present study tested the hypothesis that adolescents growing up in Children's Homes differ from adolescents growing up in a family environment in how they think about their past, present and future, in the way they make decisions about future events and rewards, and in their levels of empathy and perspective taking. The participants were 40 adolescents from Children's Homes in Budapest, Hungary, and 40 age- and gender-matched controls. Group differences were found in participants' past and present time perspectives, and girls from Children's Homes showed reduced consistency in their plans for the future. Additionally, gender differences emerged in empathy, perspective taking, and in participants' present and future time perspectives. We discuss the implications of our findings for interventions to improve the future prospects of adolescents in Children's Homes.