The Aspergillus niger annexin, anxC3.1 is constitutively expressed and is not essential for protein secretion

An annexin, anxC3.1, was isolated and characterised from the industrially important filamentous fungus Aspergillus niger. anxC3.1 is a single copy gene encoding a 506 amino acid predicted protein which contains four annexin repeats. Disruption of the anxC3.1 gene did not lead to any visible changes in phenotype, nor in the levels of secreted protein, nor specifically in glucoamylase production, suggesting no major role in secretion. anxC3.1 expression was found to be unaltered under a variety of conditions such as increased secretion, altered nitrogen source, heat shock, and decreased Ca2+ levels, indicating that anxC3.1 is constitutively expressed. This is the first reported functional characterisation of a fungal annexin.     

Production and absorption of nitric oxide gas in the nose

Some nitric oxide gas (NO) produced in thesinuses and nasal cavity is absorbed before leaving the nose. Tomeasure production and absorption, we introduced NO at differentconcentrations into one nostril while sampling the NO leaving theopposite nostril with the soft palate closed. The quantity of NO gasproduced in six normal subjects (amount leaving plus the amountabsorbed) averaged 352 nl/min and was the same at gas flows rangingfrom 8 to 347 ml/min and at 10 l/min. An absorption coefficientA was calculated by dividing theamount of NO absorbed by the concentration leaving the nose.A ranged from 17 ml/min at a nasal gasflow of 8 ml/min to an A of 24 ml/minat a nasal gas flow of 347 ml/min. The calculated rates of productionand absorption did not change when gas flow rate was increased,suggesting diffusion equilibrium. The amount of uptake of NO in thenasal mucosa can be explained by its solubility coupled with tissue andblood reactivity.

     

Long-chain formoterol analogues: an investigation into the effect of increasing amino-substituent chain length on the beta2-adrenoceptor activity

The synthesis of a series of long-chain formoterol analogues in which the terminal ether residue of the beta-phenethyl-amino-substituent has been extended beyond the methyl ether residue present in the parent compound are described. Evaluation of these analogues as beta(2)-adrenoceptor agonists was used to provide an insight into the factors controlling the magnitude and duration of receptor activation.     

Effects of diethylstilbestrol in human lymphocytes in vitro: A dose and time-dependent study on genotoxic, cytotoxic and apoptotic effects

Most of the biological, chemical or physical agents that cause cell death in certain doses and time of exposure may induce either apoptosis or necrosis. This study explores in what ways the genotoxic, cytotoxic and apoptotic effects of diethylstilbestrol (DES), a chemical agent currently used in the treatment of various types of cancer, on the human lymphocytes depend upon the dose and the exposure time. For this purpose, firstly it aims to determine in what dosages and durations of DES treatment, genotoxicity and cytotoxicity in human lymphocytes occur in vitro. Secondly, it explores the effects of DES on sister-chromatid exchanges (SCEs) and apoptosis and their relation with the nitric oxide (NO) levels. Finally, it investigates whether different dosages of DES and duration of treatment with it are correlated with each other. In so doing, we investigated the relationship among the viability, necrosis and apoptosis rates of human lymphocytes which were treated with five different DES concentrations (1, 5, 10, 15 and 20 μM) for 24, 48 and 72 h, DNA fragmentation analysis of these cells, their mean SCE values and NO levels. We concluded that 5 μM DES at 24 h is the most effective dosage that induces typical features of apoptosis in human lymphocytes. Despite the fact that there are many other studies on the effects of DES on the cancer cells, we thought it might be worth looking into the effects of DES on human lymphocytes in vitro. We meant the present study to contribute to the research done in the field of cancer treatment. (Mol Cell Biochem 276: 45–53, 2005)     

Evaluation of Oxidative Stress,Apoptosis, and Expression of MicroRNA-208a and MicroRNA-1 in Cardiovascular Patients

Background:MicroRNA expression signature and reactive oxygen species (ROS) production have been associated with the development of cardiovascular diseases (CVDs). This study aimed to evaluate oxidative stress, inflammation, apoptosis, and the expression of miRNA-208a and miRNA-1 in cardiovascular patients.Methods:The study population included four types of patients (acute coronary syndromes (ACS), myocardial infarction (MI), arrhythmia, and heart failure (HF)), with 10 people in each group, as well as a control group. Quantitative real-time PCR was performed to measure mir-208 and miR-1 expression, the mRNAs of inflammatory mediators (TNFα, iNOS/eNOS), and apoptotic factors (Bax and Bcl2). XOX, MDA, and antioxidant enzymes (CAT, SOD, and GPx) were measured by ZellBio GmbH kits by an ELISA Reader.Results:The results showed significant decreases in the activity of antioxidant enzymes (CAT, SOD, and Gpx) and a significant increase in the activity of the MDA and XOX in cardiovascular patients. Significant increases in IL-10, iNos, iNOS / eNOS, and TNF-α in cardiovascular patients were also observed. Also, a significant increase in the expression of miR-208 (HF> arrhythmia> ACS> MI) and a significant decrease in the expression of miR-1 (ACS> arrhythmia> HF> MI) were found in all four groups in cardiovascular patients.Conclusion:The results showed increases in oxidative stress, inflammation, apoptotic factors, and in the expression of miR-208a in a variety of cardiovascular patients (ACS, MI, arrhythmia, and HF). It is suggested that future studies determine the relationships that miR-1, miR-208, and oxidative stress indices have with inflammation and apoptosis.Key Words: Apoptosis, Cardiovascular diseases, Inflammation, microRNA-208a, microRNA-1, Oxidative stress     

Differential Diffusional Properties in Loose and Tight Docking Prior to Membrane Fusion

Fusion of biological membranes, although mediated by divergent proteins, is believed to follow a common pathway. It proceeds through distinct steps, including docking, merger of proximal leaflets (stalk formation), and formation of a fusion pore. However, the structure of these intermediates is difficult to study because of their short lifetime. Previously, we observed a loosely and tightly docked state preceding leaflet merger using arresting point mutations in SNARE proteins, but the nature of these states remained elusive. Here, we used interferometric scattering (iSCAT) microscopy to monitor diffusion of single vesicles across the surface of giant unilamellar vesicles (GUVs). We observed that the diffusion coefficients of arrested vesicles decreased during progression through the intermediate states. Modeling allowed for predicting the number of tethering SNARE complexes upon loose docking and the size of the interacting membrane patches upon tight docking. These results shed new light on the nature of membrane-membrane interactions immediately before fusion.     

Role of ACE and PAI-1 Polymorphisms in the Development and Progression of Diabetic Retinopathy

In the present study we determined the association of angiotensin converting enzyme (ACE) and plasminogen activator inhibitor-1 (PAI-1) gene polymorphisms with diabetic retinopathy (DR) and its sub-clinical classes in Pakistani type 2 diabetic patients. A total of 353 diabetic subjects including 160 DR and 193 diabetic non retinopathy (DNR) as well as 198 healthy controls were genotyped by allele specific polymerase chain reaction (PCR) for ACE Insertion/Deletion (ID) polymorphism, rs4646994 in intron 16 and PAI-1 4G/5G (deletion/insertion) polymorphism, rs1799768 in promoter region of the gene. To statistically assess the genotype-phenotype association, multivariate logistic regression analysis was applied to the genotype data of DR, DNR and control individuals as well as the subtypes of DR. The ACE genotype ID was found to be significantly associated with DR (p = 0.009, odds ratio (OR) 1.870 [95% confidence interval (CI) = 1.04–3.36]) and its sub-clinical class non-proliferative DR (NPDR) (p = 0.006, OR 2.250 [95% CI = 1.098–4.620]), while PAI polymorphism did not show any association with DR in the current cohort. In conclusion in Pakistani population the ACE ID polymorphism was observed to be significantly associated with DR and NPDR, but not with the severe form of the disease i.e. proliferative DR (PDR).