Detection of Bcl-2 translocation in pancreatic cancer in Iranian patients

Background and Purpose: Pancreatic cancer is one of the most leading causes of cancer-related deaths. Considering the role of Bcl-2 family gene in apoptosis that are known to be over-expressed in most cancers, this study focused on the detection of Bcl-2 translocation t(14;18) to the immunoglobulin heavy chain (IgH) that may contribute to the pathogenesis of pancreatic cancer. Â Materials and Methods: Forty-nine samples of paraffin embedded tissues (extracted from 1537 slides of 105 patients in one of the major local cancer centers) were investigated for detection of Bcl-2 translocation t(14;18) by standard PCR. Results: The Bcl-2 t(14;18) translocation was detected in 23 of all 49 patients (46.9), including 10 of 23 translocated patients (43) with break points within the mbr cluster, 11 with involvement of the mcr locus (48), and 2 in the icr locus (9). Discussion: Detection of Bcl2 translocation in pancreatic cancer is in agreement with results from other studies, and shows this rearrangement could be considered as a new treatment strategy based on apoptosis or personalized treatment. Â Keywords: apoptosis, Bcl-2 translocation, pancreatic cancer,paraffin embedded tissue, personalized treatment, polymerase chain reaction

Biofilm formation in clinical isolates of nosocomial Acinetobacter baumannii and its relationship with multidrug resistance

AbstractObjectiveTo check biofilm formation by Acinetobacter baumannii (A. baumannii) clinical isolates and show their susceptibility to different antibiotics and investigate a possible link between establishment of biofilm and multidrug resistance.MethodsThis study was performed on clinical samples collected from patients with nosocomial infections in three hospitals of Tehran. Samples were initially screened by culture and biochemical tests for the presence of different species of Acinetobacter. Identifications were further confirmed by PCR assays. Their susceptibilities to 11 antibiotics of different classes were determined by disc diffusion method according to Clinical and Laboratory Standards Institute guidelines. The ability to produce biofilm was investigated using methods: culture on Congo red agar, microtiter plate, and test tube method.ResultsFrom the overall clinical samples, 156 specimens were confirmed to contain A. baumannii. The bacteria were highly resistant to most antibiotics except polymyxin B. Of these isolates, 10.26% were able to produce biofilms as shown on Congo red agar. However, the percentage of bacteria with positive biofilm in test tube, standard microtiter plate, and modified microtiter plate assays were 48.72%, 66.66%, and 73.72%, respectively. At least 92% of the biofilm forming isolates were multidrug resistant.ConclusionsSince most of the multidrug resistant strains produce biofilm, it seems necessary to provide continuous monitoring and determination of antibiotic susceptibility of clinical A. baumannii. This would help to select the most appropriate antibiotic for treatment

Role of CYP1A1, CYP2D6, and NOS3 gene polymorphisms in idiopathic recurrent pregnancy loss in the Iranian Azeri population: A case-control study

Background: It is estimated that 1-5% of couples suffer from recurrent pregnancy loss (RPL). Recent studies have shown the effects of gene polymorphisms in RPL. Objective: The aim of this study was to evaluate 3 gene polymorphisms including rs1048943 of CYP1A1, rs28371725 of CYP2D6, and rs7830 of NOS3 in idiopathic RPL to identify their association with RPL. Materials and Methods: Blood samples were collected from 136 women with at least 2 consecutive idiopathic miscarriages (case group) and 136 women with no history of miscarriage and at least one successful pregnancy (control group) from the Iranian Azeri population. This study was carried out between April 2018-April 2020. Amplification-refractory mutation system polymerase chain reaction was used for the rs7830, rs1048943 and rs28371725 polymorphisms in order to genotype each extracted genomic DNA sample. After that, Chi-square, Fisher’s exact test and logistic regression were used to investigate whether each of these polymorphisms is associated with RPL. Results: Among these polymorphisms, only rs1048943 of CYP1A1 showed a statistically significant association with RPL in the Iranian Azeri women studied. Conclusion: Our results suggest that CYP1A1 gene polymorphisms might be associated with a reduced risk of RPL. Further studies in other populations and in the same population with a larger sample size, as well as functional genomics analyses such as gene expression analyses or epigenetic studies are required to validate our results. Key words: Recurrent pregnancy loss, Polymorphism, CYP1A1, CYP2D6, NOS3

Construction of pEGFP-ChEgTrp as DNA model for multi-epitope vaccine against Echinococcus granulosus

Background: Infection with Echinococcus granulosus causes hydatidosis in human and ruminants. With regards to the high prevalence of hydatidosis in Iran, dealing with this disease is important in terms of public health. Objective: The aim of this study was to construct pEGFP-ChEgTrp as DNA model for multiepitope vaccine against Echinococcus granulosus Methods: This experimental study was conducted in the Razi Vaccine & Serum Research Institute, Karaj in 2013. Initially, epitopes stimulating the host immune response were predicted by IEDB Database and the coding sequences were made. The sequences were amplified by PCR. The PCR products were cloned into pEGFP-N1 vector after digestion with XhoI restriction enzyme. The bacteria containing recombinant plasmid were evaluated using Colony PCR, agarose gel electrophoresis and sequencing methods. Findings: Four peptides with 10 linear epitopes were predicted in EgTrp antigen. The nucleotide sequence coding ChEgTrp was amplified by PCR using specific primers and a 270 bp fragment was obtained. This fragment was cloned into pEGFP-N1 vector and the recombinant plasmid was confirmed by Colony PCR and agarose gel electrophoresis. For final confirmation, the recombinant plasmid was sequenced and the pEGFP-ChEgTrp was constructed. Conclusion: The ChEgTrp was successfully cloned into the pEGFP-N1 vector and this plasmid can be used to design DNA vaccines. Keywords: Echinococcosis, Tropomyosin, Molecular Cloning, DNA Vaccine

Thermoacidophilic bacteria isolated from Sarcheshmeh low-grade copper ore in chalcopyrite bioleaching from mineral tailing

This research has focused on isolating and identifying different thermoacidophilic bacteria from a Sarcheshmeh low-grade copper ore and evaluating their ability of copper bioleaching from the mineral tailing. After the isolation of the bacteria, molecular identification was carried out based on the 16S rRNA gene sequences and drawing the phylogenetic tree. Then, the effect of the pH, pulp density, and composition of the media on the copper bioleaching was determined using the identified bacteria. The isolated strain (Strain SCM1) belonged to Delftia acidovorans with a 95.73% of identity. The optimal condition for the copper bioleaching was reported in a medium consisting of sulfur (10 g/L), glucose (10 g/L), yeast extract (2 g/L), and mineral tailing (5% wt/vol) at the pH of 2.00 at 50°C. Under this condition, the highest amount of copper (83%) was bioleached. It proves that the lately isolated strain can be effectively employed in the copper bioleaching process

Cloning, expression and library construction for HIV-1 Tat Protein

Background: Designing novel therapeutic agents has been a critical challenge for HIV disease.Materials and Methods: In current study a DNA sequence which was encoded the Tat protein was synthesized and inserted in pET 28 vector. Vector was cloned in BL21-DE3 E. coli and cultured in TB media. After protein expression, recombinant Tat protein was purified by NTA affinity chromatography. The Tat purified protein efficiency and confirmed by SDS-PAGE and Western blot, respectively. We were immunized the camel against HIV-1 Tat recombinant protein to made a camelid antibody library. Total RNA was extracted from camel lymphocytes and VHH fragments synthesized and amplified using RT-PCR and Nested- PCR methods by special primers.Results: The 350- 450 bp VHH gene fragment was produced by RT-PCR and Nested- PCR and extracted from agarose gel 1%. Then gel extraction was performed and pure fragments were inserted in HEN-4 vector by T4 DNA ligase.Conclusion: The library can be applied for biopanning and isolation of nanobody against HIV-1 Tat Protein. Nanobody small size may be a useful drug for treatment of HIV disease because give them the potency of the recognizing the cryptic epitopes of tat and neutralized the virus

Investigating Methylation changes of Vimentin gene in Breast Cancer

Background & Objective: Epigenetic changes are one of the most common changes in development of cancers. These changes alter the expression of certain genes that play important roles in the etiology of cancer. Increasing the expression of Vimentin gene due to epigenetic changes in Epithelial-Mesenchymal Transition(EMT) in breast carcinoma is well known. The relation between EMT and the malignancy invasion process has also been confirmed. Vimentin is the principal EMT marker that performs this task during metastasis. The purpose of this study is to investigate the methylation of the vitamin gene in a specific position. Material & methods: In this case-control study, the methylation pattern of the promoter of the Vimentin gene was evaluated in a specific site. Blood samples were taken from 30 breast cancer patients and 30 healthy individuals. DNA samples were digested with the methylation sensitive restriction enzyme. Control and treated DNA was amplified by PCR for qualitative investigation and real time PCR for quantitative analysis of methylation. Results: Using the Ct value and the%Me=100(e-0.7(ΔCt)) formula, the correlation between Vimentin hypomethylation and expression of Estrogen Receptor(ER), Progesterone Receptor(PR) and Human Epidermal growth factor Receptor2(HER2) status was checked (p-value <0.05). Conclusion: The results indicate that hypomethylated promoter of the Vim gene in the examined site correlates with ER, but does not have any correlation with PR and Her2, and hence the hypomethylation of the Vimentin gene in this position can be proposed as a molecular biomarke

Association of Y chromosome AZF region microdeletions with recurrent miscarriage in Iranian couples: A case–control study

Abstract Background Recent studies have linked recurrent pregnancy loss (RPL) to abnormalities in the sperm genome, specifically microdeletions in the azoospermia factor (AZF) region. This study investigated the potential association between Y chromosome microdeletions in the AZF region and RPL in Iranian couples. Methods The research presents a case–control study of 240 men: 120 whose partners experienced recurrent miscarriage, and 120 who had successful pregnancies without history of miscarriage. The study used semen parameters, hormone analyses, and microdeletion analysis via multiplex PCR and the YChromStrip kit. Thus, the sequence‐tagged site (STS) markers of AZFa (sY84, sY86), AZFb (sY127, sY134), and AZFc (sY254, sY255) regions were examined. Results The variations in semen parameters and sex hormone levels between cases and controls are suggest impaired testicular function in men whose partners had recurrent miscarriages (p < 0.05). Furthermore, the study revealed a negative correlation between sperm count and follicle‐stimulating hormone (FSH) level, and a positive one between sperm motility and testosterone concentration. There were no microdeletions in the control group, while the RPL group showed 20 deletions in AZFb (sY134) (16.66%) and 10 deletions each in AZFb (sY127) (8.33%) and AZFc (sY254) (8.33%). Conclusion Microdeletions in sY134 (AZFb) were significantly associated with RPL in Iranian men (p = 0.03). AZF microdeletion screening in couples with RPL can provide valuable information for ethnical genetic counseling and management of recurrent miscarriage. Further studies on larger populations or across various ethnic groups, conclusions and the inclusion of other factors like epigenetic changes explain the role of AZF microdeletions in RPL

Association study of novel single nucleotide polymorphisms of androgen receptor and estrogen receptor-α genes with male infertility in Northwest of Iran: A case-control study

Background: Observational evidence on the association of novel single nucleotide polymorphisms (SNPs) of androgen receptor (AR) and estrogen receptor-α (ER-α) genes with odds of male infertility are rare. This is particularly relevant in the Iranian population where male infertility is relatively high. Objective: This study was designed to investigate the relationship between different SNPs of these genes and male infertility among the Iranian population. Materials and Methods: The present project was a population-based, case-control study conducted on 120 idiopathic azoospermia or severe oligospermia infertile cases alongside 120 age-matched subjects enrolled as controls. Overall, 3 variants from the AR gene and 2 variants from ER-α were genotyped - AR rs137852568, AR rs137852599 and AR rs137852563, and ER-α rs796065354 and ER-α rs104893956 - using amplification refractory mutation system methods. Results: The obtained results indicated a significant association between AR rs1378525568 TT genotype as well as AR rs137852599 C allele with odds of male infertility (OR: 0.433, CI: 0.197-0.951 and OR: 0.545, CI: 0.304-0.978, respectively). Other variants of AR were not related to male infertility. A significant association was noted between predisposition polymorphism ER-α rs796065354 genotypes with male infertility. This significant association was not seen between ER-α rs104893956 and the risk of idiopathic azoospermia or severe oligospermia. Heterozygote overdominance was also observed in ESR rs796065354 but not in the other variants studied. Conclusion: Pieces of evidence were found on the association of novel polymorphisms of AR and ER-α with male infertility among the Iranian population. However, larger studies are warranted to confirm our findings. Key words: Male infertility, Androgen receptor, Estrogen receptor-α

Study of drug resistance and ompA gene existence in clinical Acinetobacter baumannii isolates

Background and Aim: Acinetobacter baumannii is one of the most important multi drug-resistant species associated with nosocomial infections. Several factors involve in resistance to drug and its pathogenicity. Of these factors, OmpA protein plays a crucial role. Therefore, the aim of current research was to assess resistance to drug and also ompA gene existence in A. baumannii isolated from clinical sources. Materials and Methods: Firstly, clinical samples were collected from Imam Khomeini, Milad and Motahari Hospitals during 2015-2016 years and the final confirmation were done by using biochemical methods. Afterwards, susceptibilities to antibiotics of different classes were determined by disc diffusion method and presence of ompA gene was checked by using PCR method and verified by sequencing. Results: The results show that of 650 clinical samples, 156 (24%) of isolates were formed of A. baumannii and mostly showed resistance to different classes of antibiotics. 92.95% of isolates were multi drug resistance (MDR) and finally 86.53% were extremely drug resistance (XDR). All of them contained ompA gene. Conclusions: Existence of multi drug resistance in most isolates as well as presence of ompA in all samples can cause bacterial virulence and drug resistance. It seems essential to provide continuous monitoring and determination of antibiotic susceptibility of clinical A. baumannii, decrease the moral and material damage caused by the bacteria