Arash Minai-Tehrani

Comparison of different probes based on labeled annexin V for detection of apoptosis

Apoptosis is a well-organized mechanism developed by eukaryotic organisms during maturation. The importance of apoptosis as a major form of cellular suicide with a causative or contributing role in a variety of diseases has become progressively obvious. A large body of evidence implicates the association of apoptosis with an arranged series of physical and biochemical alterations comprising the nucleus, cytoplasm and cell membrane. Understanding the process of apoptosis is not only important for maintaining physiological conditions, but also is critical for therapy development. In this review, we briefly outline the various ways to detect apoptosis with distinct assays and molecules, and provide an outlook on the latest imaging techniques for the visualization of phosphatidylserine (PS) externalization, with specific focus on annexin V based targeting of PS and its conjugates. Furthermore, the distinct advantages and disadvantages of each technique are reviewed. Recognition of the major benefits and drawbacks of each assay based on annexin V conjugates would allow us to use the appropriate method to detect apoptosis for instance, in a variety of diseases, such as cancer and will open novel prospects for treatment approaches.

Metabolomics: a state-of-the-art technology for better understanding of male infertility

Male factor infertility affects approximately half of the infertile couples, in spite of many years of research on male infertility treatment and diagnosis; several outstanding questions remain to be addressed. In this regard, metabolomics as a novel field of omics has been suggested to be applied for male infertility problems. A variety of terms associated with metabolite quantity and quality have been established to demonstrate mixtures of metabolites. Despite metabolomics and metabolite analyses have been around more than decades, a limited number of studies concerning male infertility have been carried out. In this review, we summarised the latest finding in metabolomics techniques and metabolomics biomarkers correlated with male infertility. The rapid progress of a variety of metabolomics platforms, such as nonoptical and optical spectroscopy, could ease separation, recognition, classification and quantification of several metabolites and their metabolic pathways. Here, we recommend that the novel biomarkers determined in the course of metabolomics analysis may stand for potential application of treatment and future clinical practice.

Untargeted metabolomic profiling of seminal plasma in nonobstructive azoospermia men: A noninvasive detection of spermatogenesis

Male factor infertility is involved in almost half of all infertile couples. Lack of the ejaculated sperm owing to testicular malfunction has been reported in 6-10% of infertile men, a condition named nonobstructive azoospermia (NOA). In this study, we investigated untargeted metabolomic profiling of the seminal plasma in NOA men using gas chromatography-mass spectrometry and advance chemometrics. In this regard, the seminal plasma fluids of 11 NOA men with TESE-negative, nine NOA men with TESE-positive and 10 fertile healthy men (as a control group) were collected. Quadratic discriminate analysis (QDA) technique was implemented on total ion chromatograms (TICs) for identification of discriminatory retention times. We developed multivariate classification models using the QDA technique. Our results revealed that the developed QDA models could predict the classes of samples using their TIC data. The receiver operating characteristic curves for these models were >0.88. After recognition of discriminatory retention times asymmetric penalized least square, evolving factor analysis, correlation optimized warping and alternating least squares strategies were applied for preprocessing and deconvolution of the overlapped chromatographic peaks. We could identify 36 discriminatory metabolites. These metabolites may be considered discriminatory biomarkers for different groups in NOA.

The use of objective oriented project planning tools for nanosafety and health concerns: a case study in nanomedicine research project

Abstract Potential human health and environmental risks associated with nanoscience research projects and their deliverables, termed nanosafety, is one of the important issues for translating research findings into commercially viable products. This paper examined the applicability of project management tools to address nanosafety in an efficient manner. Using objectives oriented project planning (OOPP) we describe a new integrated content of the problem tree, the result tree, and the logical framework approach (LFA), by modeling our nanomedicine research project entitled “Nanomedicine preparation based on antibody drug conjugate (ADC)” as a case study. As a main result of the case study, we demonstrated an LFA matrix that highlights the need to deal with nanosafety as an activity of the research project. Consequently, the activity can lead to the output, standing operating procedure (SOP), for managing the project waste disposals and its deliverables side effects. In general, such output can be concluded as an important output for all nanoscience research projects to avoid underestimating risks for their nano-objects. Moreover, this article is written in the hope of providing an easy-to-understand template of project management tools for novice nanomedicine researchers who aim to apply OOPP in the design of their research projects.

Inhibitory action of dicyclomine on lipase activity, kinetics and molecular study.

Lipase is one of the most important groups of enzymes for industry and medicine. It breaks down triacylglycerol to glycerol and fatty acids. Some bacteria use lipase to degrade the extracellular matrix of the host cells to penetrate into the tissues. Dicyclomine is a muscarinic antagonist receptor that relieves the smooth muscle spasm of the gastrointestinal tract and affects the cardiovascular system. In this research, the effect of a dicyclomine on the lipase activity of Pseudomonas aeruginosa was studied. Hanes-Woolf plot showed that the drug inhibited the enzyme by competitive inhibition. The IC50 value (60uM) and Ki (30uM) of the drug revealed that the drug bound to enzyme with high affinity. Determination of enzyme activity in various temperature showed that the maximum activity of lipase was at 60°C both in the presence and absence of the drug. Arrhenius plot determined that the activation energy of the enzyme reaction was increased in the presence of the drug. The model of binding demonstrated that the drug entered a pocket containing 10 amino acids and interacted by hydrogen bond and hydrophobic interaction and the conformational change of the enzyme after binding of the drug was confirmed by fluorescence measurement.

Inhibition of pseudoperoxiadse activity of human red blood cell hemoglobin by methocarbamol

After red blood cells lysis, hemoglobin is released to blood circulation. Hemoglobin is carried in blood by binding to haptoglobin. In normal individuals, no free hemoglobin is observed in the blood, because most of the hemoglobin is in the form of haptoglobin complex. In some diseases that are accompanied by hemolysis, the amount of released hemoglobin is higher than its complementary haptoglobin. As a result, free hemoglobin appears in the blood, which is a toxic compound for these patients and may cause renal failure, hypertensive response and risk of atherogenesis. Free hemoglobin has been determined to have peroxidase activity and considered a pseudoenzyme. In this study, the effect of methocarbamol on the peroxidase activity of human hemoglobin was investigated. Our results showed that the drug inhibited the pseudoenzyme by un-competitive inhibition. Both Km and Vmax decreased by increasing the drug concentration. Ki and IC50 values were determined as 6 and 10mM, respectively. Docking results demonstrated that methocarbamol did not attach to heme group directly. A hydrogen bond linked NH2 of carbamate group of methocarbamol to the carboxyl group of Asp126 side chain. Two other hydrogen bonds could be also observed between hydroxyl group of the drug and Ser102 and Ser133 residues of the pseudoenzyme.

Binding of Cimetidine to Balb/C Mouse Liver Catalase; Kinetics and Conformational Studies

Catalase is responsible for converting hydrogen peroxide (H2O2) into water and oxygen in cells. This enzyme has high affinity for hydrogen peroxide and can protect the cells from oxidative stress damage. Catalase is a tetramer protein and each monomer contains a heme group. Cimetidine is a histamine H2 receptor blocker which inhibits acid release from stomach and is used for gasterointestinal diseases. In this research, effect of cimetidine on the activity of liver catalase was studied and the kinetic parameters of this enzyme and its conformational changes were investigated. Cell free extract of mouse liver was used for the catalase assay. The activity of the catalase was detected in the absence and presence of cimetidine by monitoring hydrogen peroxide reduction absorbance at 240 nm. The purified enzyme was used for conformational studies by Fluorescence spectrophotometry. The data showed that cimetidine could inhibit the enzyme in a non-competitive manner. Ki and IC50 values of the drug were determined to be about 0.75 and 0.85 uM, respectively. The Arrhenius plot showed that activation energy was 6.68 and 4.77 kJ/mol in the presence and absence of the drug, respectively. Fluorescence spectrophotometry revealed that the binding of cimetidine to the purified enzyme induced hyperchromicity and red shift which determined the conformational change on the enzyme. Cimetidine could non-competitively inhibit the liver catalase with high affinity. Binding of cimetidine to the enzyme induced conformational alteration in the enzyme.