Identification of Mycobacterium species isolated from patients using high-performance liquid chromatography in Tehran during 2014-2015

Background: Non-tuberculous mycobacteria (NTM) are defined as mycobacteria other than Mycobacterium tuberculosis (MTB), which do not cause tuberculosis or leprosy. Early and precise diagnosis of NTM is particularly important for the correct epidemiological control and specific treatments. The aim of this study was to identify the mycobacterium species isolated from patients referred to hospitals in Tehran using the high-performance liquid chromatography (HPLC) method. Materials and Methods: In this cross-sectional descriptive study, a collection of isolates (n=20) was obtained from clinical specimens submitted to the Masoud Laboratory in Tehran, Iran, during 2014-2015. The strains were isolated from sputum, urine, blood, and various sterile body fluid specimens. Chromatography was conducted at a flow rate with a curvilinear gradient of methanol and methylene chloride, beginning at 98 methanol containing 2 methylene chloride and ending at 35 methanol contained in 65 methylene chloride. Results: From a total of 20 clinical isolates, 8 isolates (40) were identified as Mycobacterium abscessus, 6 isolates (30) M. tuberculosis, 3 isolates (15) M. intracellulare and 3 isolates (15) M. fortuitum. Conclusion: For the proper treatment, rapid differentiation between MTB and NTM should be performed in persons who are diagnosed with or are suspected of having infectious TB disease. So, the HPLC method can be suggested as a cost-effective, specific and reliable method for rapid identification of MTB and differentiation of NTM strain from positive cultures isolated from clinical specimens

Protective Effects of Berberine on Oxygen-Glucose Deprivation/Reperfusion on Oligodendrocyte Cell Line (OLN-93)

Background: Oligodendrocytes, the myelinating glial cells of central nervous system, are highly vulnerable to ischemic-induced excitotoxic insult, a phenomenon in which calcium overload triggers cell death. Berberine is an alkaloid extracted from medicinal herbs as Coptidis Rhizoma with several pharmacological effects like inhibition of neuronal apoptosis in cerebral ischemia. Methods: We examined the effects of berberine (0.5-4 μM) and glutamate receptors antagonists (MK-801 [10 μM] and NBQX [30 μM]) on OLN-93 cell line (a permanent immature rat oligodendrocyte) during (30, 60, 240 min) oxygen-glucose deprivation (OGD)/24 h reperfusion. The cells were cultured in 12-well plates. The cells were exposed to glucose-free medium and hypoxia in a small anaerobic chamber. Cell viability was evaluated by MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) assay. The intracellular calcium levels also were evaluated by Ca 2+ -sensitive indicator Fura-2/AM in presence or absence of berberine (2 μM) during 30 min chemical OGD by NaN3 (20 mM). Student′s t-test and ANOVA were used for statistical analysis. Results: Berberine, MK-801and NBQX significantly increased oligodendrocyte viability in all 3 time-scheduled oxygen-glucose deprivation/reperfusion. Berberine at 2 μM produced peak of protection, and increased cell viability to 83%, 77%, and 79% during 30, 60, 240 min ischemic experiments, respectively (P < 0.001). Berberine significantly attenuated intracellular Ca 2+ rise induced by chemical ischemia, and this effect of berberine was significantly stronger than MK-801 and NBQX (P < 0.001). Conclusions: We concluded that berberine protected OLN-93 oligodendrocyte against ischemic induced excitotoxic injury. Attenuation of intracellular Ca 2+ overload by berberine may be the key mechanism that saved OLN-93 from excitotoxicity damage

Preparation of a Selective L-Phenylalanine Imprinted Polymer Implicated in Patients with Phenylketonuria

Background: Molecular imprinting is a method for synthesizing polymers with structure-selective adsorption properties with applications such as, selectivity binding, drug delivery systems and anti-bodies. The present study aims at optimizing the preparation of molecularly imprinted polymer (MIP) against l-phenylalanine, in order to increase phenylalanine-binding in Enzymatic Intestinal Simulated Fluid (ESIF). Methods: The MIP for l-phenylalanine, as a water-soluble template, was successfully synthesized without derivatization. Synthesization was done by a UV polymerization method in which methacrylic acid (MAA), as a functional monomer, and ethylene glycol dimethacrylate (EGDMA), as a cross-linker, were used in the presence of five different porogenic solvents including; acetonitrile, tetrahydrofuran (THF), chloroform, toluene and dimethyl sulfoxide (DMSO). The selectivity of the MIP was examined using 19 different amino acids in human serum and was evaluated by HPLC. In addition, morphological studies were conducted using SEM. Results: The results showed that the obtained MIP with acetonitrile had the highest capacity and selectivity compared with other solvents. The data indicated that Phe-binding to MIP was significantly more than the former binding to NIP in EISF (P≤0.05). Moreover, in comparison with NIP and control group, MIP showed a better selectivity and binding for Phe. This could be used for the reduction of Phe in human serum samples of Phenylketonuria. Conclusion: Our findings suggest that the MIP against Phe prepared with acetonitrile, showed a good selectivity and binding, which caused a reduction of blood Phe concentration in enzymatic simulated intestinal fluid and human serum sample of Phenylketonuria

Commentary on prevention a possible drug-drug interaction: Is concurrent administration of orlistat and pioglitazone increase the risk of durg-induced hepatotoxicity?

Background: Drug-drug interactions (DDIs) are an emerging threat to public health and are difficult to detect. To prevent DDIs and their burden, the possible DDIs should be kept in mind. We know that the obesity predisposes to the development of insulin resistance and type 2 diabetes. Therefore, combinational uses of antiobesity drugs and glucose-lowering drugs are very common. As the hepatotoxicity of both pioglitazone (an antidiabetic drug) and orlistat (an antiobesity drug) has been shown in some cases, the aim of this study was to evaluate the interaction of pioglitazone and orlistat in human hepatocellular cell line human hepatocellular carcinoma (HepG2) cells to determine their effect on liver toxicity. Methods: Human hepatocellular carcinoma cells were treated with 25 μM Pioglitazon (Pio), 20 μM Orlistat (Orl) pioglitazone, orlistat or combination of them. The MTT assay was used to assess cell viability. Results: Pioglitazone and orlistat combination caused a loss of HepG2 cell viability. While pioglitazone (25 μM) and orliatat (20 μM) alone decreased the cell viability around 91% and 85% respectively (notsignificant, P > 0.05), the combination of these two drugs reduced the amount of viable cells to 55% which was significant when compared with each drug alone (P < 0.001). Conclusions: Revealing the significant loss of viability of HepG2 cells in the combination use of pioglitazone and orlistat indicates these two drugs should not be administered at the same time to prevent their hepatotoxic effects especially in patients with liver dysfunction

A 53 KDa Glycan Antigen of Hydatid Cyst Wall May Involve in Evasion from Host Immune System

Background: Recent studies have shown that similar host glycan antigens are expressed by helminths such as Echinococcus granulosus hydatid cysts to evade from host immune system. In this work to investigate these antigens further, immunological cross-reactivity between human sera and hydatid cyst wall antigens has been investigated. Materials and Methods: Hydatid cyst wall antigens were used in enzyme-linked immunosorbent assay and Western immunoblotting and probed with pooled sera of hydatidosis patients and healthy controls. Sodium metaperiodate treatment was used to investigate glycan antigens. Results: A band with molecular weight about 53 KDa reacted with both hydatid patients' sera and also normal human sera. It has been shown that this band was a glycan antigen. Conclusions: A 53 KDa glycan antigen of hydatid cyst wall that reacted with all human sera may have an important role for evasion from host immune system