Endothelial cell oxidant generation during K+-induced membrane depolarization

We tested the hypothesis that membrane depolarization may initiate oxidant generation in the endothelial cell. Depolarization was produced in bovine pulmonary arterial endothelial cells (BPAEC) in monolayer culture with varying external K⁺, or with glyburide (10 μM), tetraethylammonium (TEA, 10 mM), gramicidin (1 μM), or nigericin (2 μM). Evaluation of bisoxonol fluorescence of BPAEC indicated concentration-dependent depolarization by high K⁺ (2% change in fluorescence/mV change in membrane potential in the 5.9–48 mM range of K⁺) and essentially complete depolarization with glyburide. Generation of oxidants was assessed with o-phenylenediamine dihydrochloride (o-PD) oxidation in the presence of horseradish peroxidase (HRP). There was a time-dependent increase in o-PD oxidation with 24 mM K⁺, nigericin, and gramicidin over 2 hours compared with control. In 1 hour o-PD oxidation increased 2.8-fold for 24 mM and 3.7-fold for 48 mM K⁺ compared with control. Catalase reduced 24 mM K⁺-induced o-PD oxidation by 50%, while Cu/Zn-superoxide dismutase (SOD) abolished the increase. Oxidation of o-PD was reduced by 57% in the absence of HRP in the system. With K⁺ channel blockade, o-PD oxidation increased 3.8-fold with glyburide and 4.6-fold with TEA compared with control. These data indicate formation of H₂O₂ and possibly other oxidants with depolarization and suggest involvement of K⁺-channels in this process. © 1996 Wiley-Liss, Inc.     

Photoluminescence and thermoluminescence dosimetry properties of Ag/Y co-doped ZnO nanophosphor for radiation measurements

This work explores the thermoluminescence (TL) and photoluminescence (PL) properties of Ag/Y co-doped zinc oxide (ZnO) nanophosphor. The proposed dosimeter was prepared by the coprecipitation method and sintered at temperatures from 400°C to 1000°C in an air atmosphere. Raman spectroscopy was studied to investigate the structural features of this composition. The new proposed dosimeter revealed two peaks at 150°C and 175°C with a small shoulder at high temperature (225°C). The PL spectrum showed strong green emissions between 500 to 550 nm. The Raman spectrum showed many bands related to the interaction between ZnO, silver (Ag), and yttrium oxide (Y₂O₃). The rising sintering temperature enhanced the TL glow curve intensity. The Ag/Y co-doped ZnO nanophosphor showed an excellent linearity index within a dose from 1 to 4 Gy. The minimum detectable dose (MDD) of the Ag/Y co-doped ZnO nanopowder (pellets) equaled 0.518 mGy. The main TL properties were achieved in this work as follows: thermal fading (37% after 45 days at 1 and 4 Gy), optical fading (53% after 1 h and 68% after 6 h by exposure to sunlight), effective atomic number (27.6), and energy response (flat behavior from 0.1 to 1.3 MeV). Finally, the proposed material shows promising results nominated to be used for radiation measurements.     

Clinical Implications of MiR128, Angiotensin I Converting Enzyme and Vascular Endothelial Growth Factor Gene Abnormalities and Their Association with T2D

Type 2 DM (T2D) results from the interaction of the genetic and environmental risk factors. Vascular endothelial growth factor (VEGF), angiotensin I-converting enzyme (ACE), and MicroRNAs (MiRNAs) are involved in important physiological processes. Gene variations in VEGF, ACE and MiRNA genes are associated with diseases. In this study we investigated the associations of the VEGF-2578 C/A (rs699947), VEGF-2549 insertion/deletion (I/D), and ACE I/D rs4646994 and Mir128a (rs11888095) gene variations with T2D using the amplification refractory mutation system PCR (ARMS-PCR) and mutation specific PCR (MSP). We screened 122 T2D cases and 126 healthy controls (HCs) for the rs699947, and 133 T2D cases and 133 HCs for the VEGF I/D polymorphism. For the ACE I/D we screened 152 cases and 150 HCs, and we screened 129 cases and 112 HCs for the Mir128a (rs11888095). The results showed that the CA genotype of the VEGF rs699947 and D allele of the VEGF I/D polymorphisms were associated with T2D with OR =2.01, p-value = 0.011, and OR = 2.42, p-value = 0.010, respectively. The result indicated the D allele of the ACE ID was protective against T2D with OR = 0.10, p-value = 0.0001, whereas the TC genotype and the T allele of the Mir128a (rs11888095) were associated with increased risk to T2D with OR = 3.16, p-value = 0.0001, and OR = 1.68, p-value = 0.01, respectively. We conclude that the VEGF (rs699947), VEGF I/D and Mir128a (rs11888095) are potential risk loci for T2D, and that the D allele of the ACE ID polymorphism may be protective against T2D. These results help in identification and stratification for the individuals that at risk for T2D. However, future well-designed studies in different populations and with larger sample sizes are required. Moreover, studies to examine the effects of these polymorphisms on VEGF and ACE proteins are recommended.     

Analysis of Dip2B Expression in Adult Mouse Tissues Using the LacZ Reporter Gene

Disconnected (disco)-interacting protein 2 homolog B (Dip2B) is a member of the Dip2 superfamily and plays an essential role in axonal outgrowth during embryogenesis. In adults, Dip2B is highly expressed in different brain regions, as shown by in situ analysis, and may have a role in axon guidance. However, the expression and biological role of Dip2B in other somatic tissues remain unknown. To better visualize Dip2B expression and to provide insight into the roles of Dip2B during postnatal development, we used a Dip2b^{tm1a(wtsi)komp} knock-in mouse model, in which a LacZ-Neo fusion protein is expressed under Dip2b promoter and allowed Dip2B expression to be analyzed by X-gal staining. qPCR analyses showed that Dip2b mRNA was expressed in a variety of somatic tissues, including lung and kidney, in addition to brain. LacZ staining indicated that Dip2B is broadly expressed in neuronal, reproductive, and vascular tissues as well as in the kidneys, heart, liver, and lungs. Moreover, neurons and epithelial cells showed rich staining. The broad and intense patterns of Dip2B expression in adult mice provide evidence of the distribution of Dip2B in multiple locations and, thereby, its implication in numerous physiological roles.     

Therapeutic Potential of Selected Medicinal Plant Extracts against Multi-Drug Resistant Salmonella enterica serovar Typhi

Salmonella enterica serovar Typhi is Gram negative, rod shaped, facultative anaerobic bacterium, belongs to enterobacteriaceae family that causes typhoid fever in humans. This bacterium has become a super bug due to acquisition of multi drug resistance. Bacteria is transmitted through food and water contaminated with human feaces. Present study reports the screening of Adhatoda vasica, Amaranthus hybridus and Aloe barbadensis and their evaluation against multi-drug resistant Salmonella enterica serovar Typhi. Qualitative analysis of ten phytochemicals was conducted using chemical method and Gas Chromatography-Mass Spectrometry (GCMS). Antibacterial activity of plants was carried out by agar well diffusion method on Mueller Hinton agar. Total tannins, total alkaloids and total flavonoids of different parts of three plants were estimated through spectrophotometer. Total tannins content in different parts of plants was present in the given order Amaranthus hybridus leaf > Aloe barbadensis leaf > Adhatoda vasica leaf > Adhatoda vasica flower > Adhatoda vasica stem. Whereas, the order of total flavonoid concentration was Amaranthus hybridus leaf > Aloe barbadensis leaf > Adhatoda vasica leaf > Amaranthus hybridus seed. Total alkaloids have order, Adhatoda vasica leaf > Amaranthus hybridus leaf > Adhatoda vasica flower > Amaranthus hybridus seed > Aloe barbadensis leaf. Results of phytochemical analysis suggested that plants have strong profile of antioxidants, total phenolic contents and various enzymes proposing them best alternate to cure bacterial infections. GC-MS analysis further confirmed stronger phytochemical profile that can be utilized as antagonists to Salmonella enterica serovar Typhi.     

Variation in Molinia caerulea (L.) Moench, the Purple Moor Grass, in Relation to Edaphic Environments

Two diverse population of Molinia caerulea (L.) Moench, onegrowing on a Ca²⁺-rich, alkaline, Leblanc waste tip, the otheron an acid moorland, were investigated. It was anticipated thata comparative study of the morphology, growth and floweringpatterns, protein composition, specific isoenzyme and root-surfacephosphatase activities of both populations would clarify thetaxonomic status of populations of Molinia and help towardsunderstanding the ability of M. caerulea to colonize widelydiffering habitats. Both populations were identified as Moliniacaerulea caerulea. The Molinia growing on Leblanc spoil displayedcharacteristics typified by r-selected plants, i.e. displayinglarger flowers after faster development. Acid moorland plantsshowed greater vegetative development and were more typicalof K-selected types. These differences between the more vigorousshoot growth of the moorland compared to the waste tip plantspersisted under controlled conditions, irrespective of the mediain which they were grown. Root surface phosphatase activityshowed a plastic response to edaphic pH which may contributeto the success of M. caerulea caerulea in colonizing diversehabitats.Copyright 1995, 1999 Academic Press Molinia, edaphic extremes, variation, morphometry, enzyme activities, plasticity, colonization, taxonomy     

A comparative study among dietary supplementations of antibiotic,grape seed and chamomile oils on growth performance and carcass properties of growing rabbits

The main objective of this study was to evaluate the effect of chamomile oil (Ch), grape seed oil (GS), their mixture and antibiotic (colistin) (AN) as feed addetives on the productivity of growing rabbits as well as in vitro study to evaluate the antimicrobial activity of both Ch and GS oils. To achive this objective, a total of 96 New Zealand (NZW) weaned rabbits, 5 weeks-old were randomly allotted into eight groups. Rabbits were kept under observation for eight weeks and the trial ended at thirteen weeks-old. The experimental treatments were: 1) Basal diet (BD); 2) BD + antibiotic; 3) BD + 0.5 ml GS/ kg diet; 4) BD + 1.0 ml GS/ kg diet; 5) BD + 1.5 ml GS/ kg diet; 6) BD + 0.5 ml Ch/ kg diet; 7) BD + 1.0 ml Ch/ kg diet and 8) BD + 1.5 Ch/ kg diet. Live body weight (LBW) was markedly elevated (p < 0.05) in groups fed on ration included feed additives compared with the control at weeks 9 and 13 of age. Cumulative body weight gain (BWG) and feed intake (FI) increased (p < 0.05) throughout 5–9 and 5–13 weeks of age in rabbits fed rations plus the studied additives. Feed conversion ratio (FCR) was insignificantly altered by dietary feed additives. Spleen and intestine relative weights reduced (p < 0.05) in groups treated with different studied additives. In view of the experiment finings, it could be concluded that dietary supplementation of GS and Ch have a positive impact on the productivity of growing rabbits than that of the control and antibiotic-treated groups.     

热解纤维素菌属F32中乙酰木聚糖酯酶Axe7的表征

【目的】从嗜热厌氧微生物热解纤维素菌属F32(Caldicellulosiruptor sp.F32)菌株中鉴定出可水解木聚糖侧链乙酰基团的脂酶。【方法】通过基因组序列注释、比对以及蛋白结构预测的方法,发现一个潜在的脂酶7家族的(CE-7)乙酰木聚糖脂酶Axe7。利用基因克隆、质粒构建以及在大肠杆菌中表达目标蛋白并纯化等实验方法,获得了该酶的重组蛋白。【结果】以4-甲基乙酸伞形酯(4-Methylumbelliferyl-acetate)作为底物时,该酶的最适反应p H在6.5-7.0之间,最适反应温度为85°C,在最适的温度和p H条件下,Axe7活性半衰期(Half-life)超过4 h。在不同金属离子(1.5 mmol/L)存在下,Axe7活性可保持为最适反应酶活的(66.3±4.6)%-(95.7±2.3)%之间,说明金属离子对其酶活有一定的影响。通过测定酶动力学发现Axe7的Km和kcat值分别为0.39 mmol/L和66.95 s-1。【结论】从高温厌氧微生物中发现并表征一个热稳定性良好的乙酰木聚糖脂酶,为木质纤维素的高温糖化和生物炼制提供了一个可工业化的潜在选择。