The Usefulness of DNA Sequencing After Extraction by Whatman FTA Filter Matrix Technology and Phenotypic Tests for Differentiation of Candida albicans and Candida dubliniensis

Since C. dubliniensis is similar to C. albicans phenotypically, it can be misidentified as C. albicans. We aimed to investigate the prevalence of C. dubliniensis among isolates previously identified as C. albicans in our stocks and to compare the phenotypic methods and DNA sequencing of D1/D2 region on the ribosomal large subunit (rLSU) gene. A total of 850 isolates included in this study. Phenotypic identification was performed based on germ tube formation, chlamydospore production, colony colors on chromogenic agar, inability of growth at 45 °C and growth on hypertonic Sabouraud dextrose agar. Eighty isolates compatible with C. dubliniensis by at least one phenotypic test were included in the sequence analysis. Nested PCR amplification of D1/D2 region of the rLSU gene was performed after the fungal DNA extraction by Whatman FTA filter paper technology. The sequencing analysis of PCR products carried out by an automated capillary gel electrophoresis device. The rate of C. dubliniensis was 2.35 % (n = 20) among isolates previously described as C. albicans. Consequently, none of the phenotypic tests provided satisfactory performance alone in our study, and molecular methods required special equipment and high cost. Thus, at least two phenotypic methods can be used for identification of C. dubliniensis, and molecular methods can be used for confirmation.     

Antioxidant effect of carnosine treatment on renal oxidative stress in streptozotocin-induced diabetic rats

Nitric oxide (NO) plays a significant role in the development of diabetic nephropathy. We investigated the effects of an antioxidant, carnosine, on streptozotocin (STZ)-induced renal injury in diabetic rats. We used four groups of eight rats: group 1, control; group 2, carnosine treated; group 3, untreated diabetic; group 4, carnosine treated diabetic. Kidneys were removed and processed, and sections were stained with periodic acid-Schiff (PAS) and subjected to eNOS immunohistochemistry. Examination by light microscopy revealed degenerated glomeruli, thickened basement membrane and glycogen accumulation in the tubules of diabetic kidneys. Carnosine treatment prevented the renal morphological damage caused by diabetes. Moreover, administration of carnosine decreased somewhat the oxidative damage of diabetic nephropathy. Appropriate doses of carnosine might be a useful therapeutic option to reduce oxidative stress and associated renal injury in diabetes mellitus.     

Synthesis of some novel pyridine compounds containing bis‐1,2,4‐triazole/thiosemicarbazide moiety and investigation of their antioxidant properties,carbonic anhydrase,and acetylcholinesterase enzymes inhibition profiles

Some novel derivatives of thiosemicarbazide and 1,2,4‐triazole‐3‐thiol were synthesized and evaluated for their biological activities. The title compounds were prepared starting from readily available pyridine‐2,5‐dicarboxylic acid. The reaction carboxylic acid with absolute ethanol afforded the corresponding dimethyl pyridine‐2,5‐dicarboxylate ( 1 ). The reaction of dimethyl‐2,5‐pyridinedicarboxylate ( 1 ) with hydrazine hydrate good yielded pyridine‐2,5‐dicarbohydrazide ( 2 ). Refluxing compound 2 with alkyl/aryl isothiocyanate derivatives for 3–8 h afforded 1,4‐disubstituted thiosemicarbazides ( 3a–e ). Base‐catalyzed intra‐molecular dehydrative cyclization of these intermediates furnished the 4,5‐disubstituted bis‐mercaptotriazoles ( 4a–e ) in good yield (85%–95%). Among the target compounds, 2,2′‐(pyridine‐2,5‐diyldicarbonyl)bis[N‐(p‐methoxyphenyl)hydrazinecarbothioamide] ( 3c ) showed very high activity with value of 72.93% against 1,1‐diphenyl‐2‐picrylhydrazyl free radical at the concentration of 25 μg/mL. The inhibitory effects of the target compounds against acetylcholinesterase (AChE), hCA I, and II were studied. AChE, cytosolic hCA I and II isoforms were potently inhibited by synthesized these derivatives with K_is in the range of 3.07 ± 0.76–87.26 ± 29.25 nM against AChE, in the range of 1.47 ± 0.37–10.06 ± 2.96 nM against hCA I, and in the range of 3.55 ± 0.57–7.66 ± 2.06 nM against hCA II, respectively.     

Molecular genetic diversity and association mapping of morphine content and agronomic traits in Turkish opium poppy (<Emphasis Type="Italic">Papaver somniferum</Emphasis>) germplasm

As the sole plant source of many potent alkaloids, opium poppy (Papaver somniferum L.) is an important medicinal crop. Nevertheless, few studies have characterized opium poppy germplasm with crop-specific molecular markers. Because Turkey is a diversity center for opium poppy, Turkish germplasm is a valuable genetic resource for association mapping studies aimed at identifying QTLs controlling morphine content and agronomic traits. In this study, the morphological diversity and molecular diversity of 103 Turkish opium poppy landraces and 15 cultivars were analyzed. Potentially useful morphological variation was observed for morphine content, plant height, and capsule index. However, the landraces exhibited limited breeding potential for stigma number, and seed and straw yields. Both morphological and molecular analyses showed distinct clustering of cultivars and landraces. In addition, a total of 164 SSR and 367 AFLP polymorphic loci were applied to an opium poppy association mapping panel composed of 95 opium poppy landraces which were grown for two seasons. One SSR and three AFLP loci were found to be significantly associated with morphine content (P < 0.01 and LD value (r ²) = 0.10–0.32), and six SSR and 14 AFLP loci were significantly associated with five agronomic traits (plant height, stigma number, capsule index, and seed and straw yields) (P < 0.01 and LD value (r ²) = 0.08–0.35). This is the first report of association mapping in this crop. The identified markers provide initial information for marker-assisted selection of important traits in opium poppy breeding.     

Effects of dietary mannanoligosaccharide on performance, some blood parameters, IgG levels and antibody response of lambs to parenterally administered E. coli O157:H7

Forty-eight male lambs were used to evaluate the effect of dietary supplementation of mannanoligosaccharide (MOS) with or without parenteral Escherichia coli injection on their growth performance, feed conversion efficiency, blood metabolites, total serum immunoglobulin G (IgG) levels and antibody response. Lambs were randomly assigned to four groups of 12 animals each. In groups C (control) and CE (E. coli challenged), animals were fed commercial concentrate pellets and hay (50:50), and in groups M (MOS) and ME (MOS + E. coli challenged), animals were fed commercial concentrate pellets including MOS at 0.2% and hay (50:50). At day 15 and 30, animals in groups CE and ME were injected subcutaneously with 1 ml of phosphate buffered saline (PBS) suspension containing 10(6) cfu of heat inactivated non-toxigenic E. coli O157:H7, while animals in C and M groups were injected subcutaneously with 1 ml of PBS. The experimental period was 45 days. Data indicated that body weight of lambs at the end of the study were statistically non-significant among the groups. Blood metabolites, i.e. total protein, albumin, calcium and phosphorus concentrations were not affected significantly by MOS supplementation. However, administration E. coli lowered (p < 0.05) total protein, albumin and calcium concentrations in the serum on day 30. The IgG level was not different between groups. However, on day 45, the total IgG level was found to be higher (p < 0.05) in lambs that had received MOS and E. coli than in other groups. Application of MOS did not have any effect on the antibody response to E. coli as OD values.     

The Effects of Boron on Arsenic‐Induced Lipid Peroxidation and Antioxidant Status in Male and Female Rats

The aim of the present study was to investigate the possible protective effects of boron, an antioxidant agent, against arsenic‐induced oxidative stress in male and female rats. In total, 42 Wistar albino male and female rats were divided into three equal groups: The animals in the control group were given normal drinking water, the second group was given drinking water with 100 mg/L arsenic, and the third group was orally administered drinking water with 100 mg/kg boron together with arsenic. At the end of the 28‐day experiment, arsenic increased lipid peroxidation and damage in the tissues of rats. However, boron treatment reversed this arsenic‐induced lipid peroxidation and activities of antioxidant enzymes in rats. Moreover, boron exhibited a protective action against arsenic‐induced histopathological changes in the tissues of rats. In conclusion, boron was found to be effective in protecting rats against arsenic‐induced lipid peroxidation by enhancing antioxidant defense mechanisms.     

Quantitative proteomics reveals differential regulation of protein expression in recipient myocardium after trilineage cardiovascular cell transplantation

Intramyocardial transplantation of cardiomyocytes (CMs), endothelial cells (ECs), and smooth muscle cells (SMCs) derived from human induced pluripotent stem cells (hiPSCs) has beneficial effects on the post‐infarction heart. However, the mechanisms underlying the functional improvements remain undefined. We employed large‐scale label‐free quantitative proteomics to identify proteins that were differentially regulated following cellular transplantation in a swine model of myocardial infarction (MI). We identified 22 proteins that were significantly up‐regulated after trilineage cell transplantation compared to both MI and Sham groups. Among them, 12 proteins, including adenylyl cyclase‐associated protein 1 and tropomodulin‐1, are associated with positive regulation of muscular contraction whereas 11 proteins, such as desmoplakin and zyxin, are involved in embryonic and muscular development and regeneration. Moreover, we identified 21 proteins up‐regulated and another 21 down‐regulated in MI, but reversed after trilineage cell transplantation. Proteins up‐regulated after MI but reversed by transplantation are related to fibrosis and apoptosis. Conversely, proteins down‐regulated in MI but restored after cell therapy are regulators of protein nitrosylation. Our results show that the functionally beneficial effects of trilineage cell therapy are accompanied by differential regulation of protein expression in the recipient myocardium, which may contribute to the improved cardiac function.     

Reactive oxygen species and ischemic cerebrovascular disease

Stroke is an emerging major health problem often resulting in death or disability. Hyperlipidemia, high blood pressure and diabetes are well established risk factors. Endothelial dysfunction associated with these risk factors underlies pathological processes leading to atherogenesis and cerebral ischemic injury. While mechanisms of disease are complex, endothelial dysfunction involves decreased nitric oxide (NO) and elevated levels of reactive oxygen species (ROS). At physiological levels, ROS participate in regulation of cellular metabolism. However, when ROS increase to toxic levels through imbalance of production and neutralization by antioxidant enzymes, they cause cellular injury in the form of lipid peroxidation, protein oxidation and DNA damage. Central nervous system cells are more vulnerable to ROS toxicity due to their inherent higher oxidative metabolism and less antioxidant enzymes, as well as higher content of membranous fatty acids. During ischemic stroke, ROS concentration rises from normal low levels to a peak point during reperfusion possibly underlying apoptosis or cellular necrosis. Clinical trials and animal studies have shown that natural compounds can reduce oxidative stress due to excessive ROS through their antioxidant properties. With further study, we may be able to incorporate these compounds into clinical use with potential efficacy for both the treatment and prevention of stroke.