Comparative evaluation of sequence analysis of 16S rRNA and rpoB genes for identification of aquatic mycobacteria

The nucleotide sequences of partial 16S rRNA and bacterial RNA polymerase ß-subunit (rpoB) genes for 57 mycobacterial strains were determined. Compared to the 16S rRNA gene sequences, variable regions were scattered along the whole fragment sequence, indicating that therpoBgene is more polymorphic. Unlike 16S rRNA sequences, species variation was observed within M. fortuitum strains. The topology of therpoB-based phylogenetic tree was almost the same as that of the 16S rRNA sequence analysis. These results suggest that therpoBgene is a highly conserved gene, and taxonomical studies based on this gene may be comparable with similar studies based on the 16S rRNA gene. The overall mean distance forrpoB-gene?based sequences was 2.5 times greater than that of the 16S rRNA gene for all 57 mycobacterial strains examined. However, some slowly growing mycobacteria could not be differentiated based onrpoBgene sequences. Moreover, a bootstrap value above 70% was observed for 13 nodes, while this value was 14 nodes in the case of 16S rRNA sequences. To the best of our knowledge, this is the first investigation evaluating the use of 16S rRNA andrpoBsequence analyses for identification of aquatic mycobacteria obtained from diverse geographical locations

Selective Toxicity of Standardized Extracts of Persian Gulf Sponge ( Irciniamutans) on Skin Cells and Mitochondria isolated from Melanoma induced mouse

Melanoma is an aggressive and highly lethal cancer with poor prognosis and resistance to current treatments. Apoptosis signaling is believed to be suppressed in melanoma. Evidence suggests that compounds isolated from marine sponges have anti-cancer properties. This study was designed to evaluate the apoptotic effect of methanolic, diethyl ether, and n-hexane extracts of Irciniamutans (I.mutans) on skin mitochondria isolated from mice animal models of melanoma. Mitochondria were isolated by differential centrifugation. According to our results, methanolic, diethyl ether, and n-hexane extracts of I.mutans raised the reactive oxygen species (ROS) level only in the cancerous skin mitochondria group. Furthermore, methanolic, diethyl ether, and n-hexane extracts induced swelling in the mitochondria, decreased the mitochondrial membrane potential (MMP) and the release of cytochrome c from the mitochondria. Based on these results, the potentially bioactive compounds found in I.mutans render the sea sponge as a strong candidate for further researches in molecular identification and confirmatory in-vivo researches

The effect of some polymorphisms in vitamin D receptor gene in menopausal women with osteoporosis

Introduction: Vitamin D receptor gene is one of candidate genes related to osteoporosis expansion. The association of ApaI, TaqI, and BsmI polymorphisms in vitamin D receptor gene with bone metabolism and density has been already demonstrated. Aim: This study was conducted to investigate the association between the ApaI, TaqI and BsmI polymorphisms and bone density. This study was descriptive-analytical study. Centers for bone density measurement in southwestern Iran. Materials and Methods: In this descriptive-analytical study, 200 participants aged 45- and above 45-year-old women referring the centers of bone density measurement participated. The bone density of femoral neck and lumbar vertebrae was measured using dual-energy X-ray absorptiometry method. Based on t-score, the participants were assigned into patients (n=130) and healthy individuals (n=70). Different genotypes of ApaI (AA/Aa/aa), TaqI (TT/Tt/tt), and BsmI (BB/Bb/bb) were determined by PCR-RFLP. The data on bone density and PCR-RFLP were analysed by chi-square and ANOVA. Also, triad combination of the genotypes was statistically analysed. For each genotype combination, chi-square was run between the patients and control group and p-value was calculated. Results: No significant association was seen between ApaI polymorphism and bone density (p>0.05). TaqI and BsmI polymorphisms had a significant association with femoral neck’s bone density (p0.05). Patients with homozygous dominant TT genotype had the least bone density in femoral neck compared to other genotypes. Lumbar vertebrae’s bone density was similar in three TaqI genotypes. The patients with homozygous recessive bb genotype had the least bone density in femoral neck and lumbar vertebrae compared to other genotypes. Conclusion: TaqI and BsmI polymorphisms could be desirable markers in diagnosis of women at risk of osteoporosis in the studied region in Iran. Therefore, these women will receive suitable medical treatment at proper tim

Toxicity Evaluation of 6-Mercaptopurine Using Accelerated Cytotoxicity Mechanism Screening (ACMS) techniques

6- Mercaptopurine (6-MP) is widely used in clinic as an immunosuppressive for treatment of acute lymphocytic leukemia, Crohn's disease, and ulcerative colitis with documented unpredictable hepatotoxicity. The potential molecular cytotoxic mechanisms of 6-MP against isolated rat hepatocytes were searched in this study using ―Accelerated Cytotoxicity Mechanism Screening (ACMS)‖ techniques. The concentration of 6-MP required to cause 50% cytotoxicity in 2 hour at 37∘C was detected to be 400 μM. A significant increase in 6-MP induced cytotoxicity and reactive oxygen species (ROS) formation, % mitochondrial membrane potential (MMP), lysosomal damage were observed. The addition of chloroquine (lysosomotropic agent), L-carnitine (inhibitor of membrane permeability transition (MPT), Diphenyleneiodonium (DPI) as an inhibitor of production of superoxide, and H2O2 by mitochondria and Dimethyl sulfoxide (DMSO) as a radical scavenger decreased 6-MP-induced cytotoxicity, ROS formation, collapse of MMP, and lysosomal damage. Results from this study suggest that 6-MP -induced cytotoxicity in isolated rat hepatocytes due to ROS formation, mitochondrial and lysosomal damages that resulted in crosstalk toxicity between mitochondrial and lysosomal damage and finally cell death

Toxicity of popular NSAIDs on hearth mitochondria

Introduction: Several chemical compounds and drugs have been known to directly or indirectly modulate cardiac mitochondrial function, which can account for their cardiotoxic and arrhythmic properties. Non-steroidal anti-inflammatory drugs (NSAIDs) are most prescribed drugs in human and veterinary medicine that provide anti-inflammatory, antipyretic, analgesic, antispasmodic and anticoagulant effects. Methods and Results: Rat heart mitochondria were obtained by differential ultracentrifugation and incubated with different concentrations of highly prescribed NSAIDs (diclofenac, naproxen, celecoxib). Our results showed that NSAIDs (diclofenac, naproxen, celecoxib) induced a rise in cardiac mitochondrial reactive oxygen species (ROS) formation, lipid peroxidation, and mitochondrial membrane potential (MMP) collapse before mitochondrial swelling ensued on isolated rat heart mitochondria. Disturbance in oxidative phosphorylation was also confirmed by the decrease in ATP concentration in the NSAIDs (diclofenac, naproxen, celecoxib)-treated heart mitochondria. In addition, the collapse of MMP and mitochondrial swelling produced the release of cytochrome c via outer membrane rupture or mitochondrial permeability transition (MPT) pore opening. Conclusions: Our results suggested that NSAIDs (diclofenac, naproxen, celecoxib)-induced toxicity in heart tissue is the result of disruptive effect on mitochondrial respiratory chain that leads to ROS formation, lipid peroxidation, MMP decline, and cytochrome c expulsion which results in apoptosis signaling and cell loss in heart myocardial tissue

Calcitonin receptor AluI (rs1801197) and TaqI calcitonin genes polymorphism in 45-and over 45-year-old women and their association with bone density

Purpose: Calcitonin receptor gene has also a polymorphism which is associated with bone mass density. This study evaluates the association between calcitonin receptor AluI (rs1801197) and Taq1 calcitonin genes polymorphism with bone density rate. Methods: In this descriptive-analytical study in 2013 in southwestern Iran, 200 blood samples, per the Cochran sample size formula, were taken from women aged 45 and older. DNA was extracted from the samples using the phenol- chloroform method and the genomic fragments in question were proliferated using the polymerase chain reaction (PCR) method. Results: The genotypic distribution of polymorphism AluI for TT, TC, and CC genotypes in control group was 31.4%, 38.6%, and 30% and in patients 25.4%, 55.4%, and 19.2%, respectively. There was no significant difference in polymorphism AluI between patients and control group and no significant association was found between this gene and bone density rate (P > 0.05). All patients and the individuals in the control group exhibited tt genotype for TaqI calcitonin gene and no significant association was found between these participants and osteoporosis. Conclusion: There was no association between two polymorphisms and osteoporosis, and between polymorphism of these two genes and osteoporosis development rate in the participants. © 2016 Morteza Dehghan, Razieh Pourahmad-Jaktaji, and Zarghampoor Farzaneh

Investigation of the presence of G354A (Cys87Tyr) mutation in osteoprotegerin gene in women with osteoporosis in Chaharmahal and Bakhtiari province

Background and aims: Osteoprotegerin (OPG) is a competitive inhibitor of the differentiation and activity of osteoclasts, which inhibits the final stages of osteoclast formation and induces its apoptosis. In addition, OPG is considered as one of the most important candidate genes in the pathogenesis of bone diseases such as osteoporosis and idiopathic hyperphosphatasia. The G354A (Cys87Tyr) mutation in the OPG gene leads to idiopathic hyperphosphatasia. This mutation is probably related to osteoporosis. The purpose of this study was to investigate the presence of G354A (Cys87Tyr) in women with osteoporosis in Chaharmahal and Bakhtiari province. Methods: In this descriptive-analytical study, the bone mineral density (BMD) of the femoral neck and lumbar spine of women referring to Shahrekord bone densitometry centers was measured by the X-ray absorptiometry technique in 2013-2014. Based on T-scores, people with osteoporosis were identified and 70 patients were enrolled in the study after receiving their consent. Finally, DNA was extracted from blood samples, amplified by polymerase chain reaction (PCR) technique, and sequenced by DNA sequencing method. Results: After DNA extraction from the blood, the quality and quantity were determined by gel electrophoresis and spectrophotometry, respectively. Then, the gene was amplified by the PCR method and the product was detected by gel electrophoresis, followed by sequencing the samples to investigate the presence of the mutation. Eventually, genotypes associated with Cys87Tyr mutation were not observed in the studied population. Conclusion: In the present study, the G354A (Cys87Tyr) mutation associated with idiopathic hyperphosphatasia was not found in women with osteoporosis. Keywords: G354A mutation, OPG gene, Osteoporosi

Enrichment of Probiotic Yogurt with Broccoli Sprout Extract and its Effect on Helicobacter pylori

Background and Objective: Antibiotic consumption is the main way to cure infection induced by Helicobacter pylori. On the other hand, antibiotics have side effects on human body. So, finding an efficient way to replace antibiotic consumption seems necessary. The aim of this study was to investigate the effect of broccoli sprout extract on the viability of probiotic bacteria and yogurt’s physicochemical properties, and examine the synergistic effect of this extract with probiotics on Helicobacter pylori growth inhibition.Material and Methods: Four levels of broccoli sprout extract (22.5, 45, 90 and 180 mg ml-1) were prepared and their effect on probiotic yogurt samples was examined. Moreover, their anti- Helicobacter pylori effect was determined.Results and Conclusion: The research results revealed that Broccoli sprout extract did not have any inhibitory effect on Bifidobacterium lactis and Lactobacillus acidophilus. The variations in acidity of the samples were not significant during storage. A positive correlation was observed between broccoli sprout extract concentration and syneresis. The findings showed the synergistic effect of broccoli sprout extract and probiotics on Helicobacter pylori growth inhibition. Therefore, using broccoli sprout extract and probiotic bacteria, we can produce a yogurt that is effective on the growth inhibition of Helicobacter pylori.Conflict of interest: The authors declare that there is no conflict of interest

Can Physical Activity Patterns before and during Pregnancy and Anxiety be Related to Preterm Birth?

AbstractIntroduction: Changes of physical activity before and during three trimesters of pregnancy known as pattern of physical activity and anxiety during pregnancy can be two concerns of pregnant women about preterm birth which require clarification. So, this retrospective study aimed to assess the effect of pattern of physical activity, and anxiety on preterm birth in Iranian pregnant women.Methods: This study was a kind of descriptive correlation which was performed retrospectively. Participants of this study (2019-2020) included 118 pregnant women with preterm (n=62) and term (n=56) birth who participated in the study voluntarily and were selected according to the study inclusion criteria. The energy cost of physical activity before and during three trimesters of pregnancy, and anxiety level were recorded through interview using pregnancy physical activity (PPAQ) and Hospital Anxiety and Depression Rating Scale Anxiety (HADS) questionnaires respectively.Results: There was no significant difference between preterm and term birth groups regarding total physical activity, and sedentary behavior energy expenditure before and during pregnancy as well as anxiety during pregnancy(P>0.05). Physical activity pattern was not significantly different between the two groups (P>0.05). However, physical activity before pregnancy was significantly higher than all trimesters of pregnancy in both groups of the study (P<0.001). In two groups of preterm and term deliver, physical activity reduced in the third compared to the second trimester of pregnancy similarly.Conclusions: Physical activity reduced during pregnancy in preterm and term birth women similarly. In this study, physical activity, sedentary behavior and anxiety during pregnancy and before it were not effective on the pre-term birth