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Background and objectives: HIV-1 Nef and Vpr antigens have been described as suitable candidates for therapeutic HIV vaccine development. The aim of this study was to generate Nef-Vpr fusion gene construct and to clone the construct into pET-23a (+), a prokaryotic expression vector.
Methods: HIV-1 Nef and Vpr genes were PCR-amplified from the pNL4-3 plasmid using specific primers and Pfu DNA polymerase. Results of PCR amplification were visualized by electrophoresis on 0.8% agarose gel. At first, the amplified Nef fragment was cloned into NheI and BamHI restriction sites of pET-23a expression vector. Next, cloning of Vpr gene was performed into BamHI and HindIII restriction sites of the pET-23a-Nef vector. Finally, purity of the recombinant pET-23-Nef-Vpr construct was determined by NanoDrop spectrophotometry.
Results: PCR amplification of Nef and Vpr genes was confirmed by detection of ~ 620 bp and ~ 291 bp bands, respectively. Cloning of the Nef-Vpr construct into the vector was confirmed by detection of a ~ 911 bp fragment following enzymatic digestion with NheI and HindIII and sequencing.
Conclusion: The successful construction of recombinant fusion plasmid encoding a chimeric Nef-Vpr gene was performed in a prokaryotic expression vector for development of HIV-1 recombinant protein vaccine in near future.

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Background and objectives: Antibiotic resistance is a global health challenge that affects both individuals and the health system in many ways. The aim of this study was to evaluate the antibiotic resistance pattern in isolates from patients admitted to the intensive care unit (ICU) of a hospital in Qazvin, Iran.
Methods: This descriptive and retrospective study was performed on urine and blood samples collected from 1318 ICU patients in the Velayat Hospital of Qazvin (Iran) during 2017-2019. Data were collected from patients’ medical records. All statistical analyses were performed using SPSS software (version 25).
Results: Based on the findings, 65.2% of the samples were related to urinary tract infections and 34.7% to bloodstream infections. Escherichia coli (68.6%) and Stenotrophomonas (41.0%) were the most common bacteria isolated from urinary tract infections and bloodstream infections, respectively. Moreover, the rate of antibiotic resistance was higher among Acinetobacter, Escherichia coli, Stenotrophomonas, Enterococcus and Pseudomonas isolates.
Conclusion: The rate of drug resistance in isolates from ICU patients is alarmingly high and requires immediate attention. It is recommended to modify antibiotic prescriptions in the hospital based on the results of antibiotic resistance pattern, particularly for treatment of infections caused by E. coli and Stenotrophomonas.

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Coma is a state of prolonged unconsciousness. Some coma cases result from inherited disorders such as fatty-acid β-oxidation disorder, acute intermittent porphyria (due to mutations in genes CPT I, CPTII and ACADM), urea cycle defects (due to mutation in OTC gene), organic acidurias, mitochondrial diseases and familial hemiplegic migraine (due to mutations in CACNA1A, ATP1A2 and SCN1A). The evaluation of familial cases of coma or sporadic coma can be performed using next generation sequencing (NGS), a high-throughput  sequencing technique that can sequence an entire genome in a single reaction. This technique has been widely applied in the genetic diagnosis of diseases. In this review, we describe some genes associated with coma or recurrent coma and discuss the role of NGS in detection of these genes.  

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Background and objectives: Previous investigations have shown that physical activity can improve insulin sensitivity and body composition by reducing the concentration of inflammatory biomarkers. The study aimed at evaluating effects of eight weeks of resistance training and high-intensity interval training on adropin, blood glucose markers, and body composition in overweight females.
Methods: sixteen overweight females (mean age: 30 ± 4.3 years and body mass index= 29 ± 2.6 kg/m²) were randomly assigned to a resistance training group (n=8) and a high-intensity interval training group (n=8). Participants in both groups trained three times a week for eight weeks. Body composition and serum level of blood markers were determined at baseline and after the last training session.
Results: Body mass, body fat percentage, and waist-hip ratio decreased significantly in both groups (p<0.05). VO_2max significantly increased in both groups, while the changes in the resistance training group were greater than in the high-intensity interval training group (p<0.05). Insulin and HOMA-IR concentrations decreased significantly in the resistance training group (p<0.05).
Conclusion: Eight weeks of both training procedures could significantly decrease body composition markers. However, the training duration was not sufficient to alter fating blood sugar or adropin concentrations.

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Background and objectives: Recycled polyethylene terephthalate (RPET) nanofibers have become an important part of human life, with a continuous increase in their production and consumption. Herein, the antibacterial activity of nickel nanoparticles/recycled polyethylene terephthalate nanofibers (NiNP/RPET NF web) was evaluated by analyzing alginate expression in Pseudomonas aeruginosa, as an opportunistic microorganism.
Methods: NiNPs were synthesized and NiNP/RPET NF was produced by adding 25 μg/ml of NiNP to 10% solutions of RPET at a weight ratio of 3%. After exposing P. aeruginosa (PA01) to NiNP/RPET NF, the biofilm-forming capacity was determined and real-time PCR was performed to measure algD expression.
Results: Treatment with 25 μg/ml of NiNP/RPET NF reduced growth of P. aeruginosa on Mueller Hinton agar but did not result in complete inhibition. The biofilm optical density (550 nm) was 0.464 ± 0.021 after treatment with NiNP/RPET NF and 0.082± 0.011 in the absence of NiNP/RPET NF. This indicates the significant reduction of biofilm formation after exposure to NiNP/RPET NF (p=0.01). In addition, a 0.6-fold (p=0.03) reduction in alginate expression was detected by real-time quantitative real-time PCR.
Conclusion: Our results indicate the potential of NiNP/RPET NF for application in nano-based antibacterial medical systems.

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Background and objectives: Cutaneous leishmaniasis is endemic in most areas of Iran, and the diagnosis of its species is essential for controlling the disease. Leishmania major is the causative agent for cutaneous leishmaniasis in humans. Molecular methods are generally more sensitive than microscopic methods. The present study aimed to use a polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA) technique for detecting live L. major from wounds of patients with cutaneous leishmaniasis.
Methods: In the present study, a standard strain of L. major promastigotes was used as the positive control for purification of DNA. The Novy–MacNeal–Nicolle and RPMI-1640 media were used for reproduction of parasites. DNA was isolated from specimens taken from 35 patients with suspected cutaneous leishmaniasis whose disease was confirmed by direct smear method. The PCR-ELISA technique was later applied by using the standard strain, patient specimens, and primers specific for the 18s rRNA.
Results: Out of 35 patients, 17 (48.6%) were male and 18 (51.4%) were female. In addition, 8.6% of the patients lived in the Gonbad-e Kavus County, while all patients had been infected in villages around Gonbad-e Kavus. Of 35 patients with confirmed cutaneous leishmaniasis according to the direct smear method, 31 patients (86.31%) had leishmaniasis based on the PCR method and the PCR-ELISA methods.
Conclusion: Based on the results, the PCR-ELISA method is more sensitive and accurate for detecting L. major.

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Background and objectives: Enterococcus faecium is a normal flora of gut microbiota. This opportunistic pathogen has attracted much attention due to its multidrug resistance and ability to survive in hostile environments. Various molecular typing methods such as pulsed-field gel electrophoresis or ribotyping have been developed for clinical and epidemiological investigation of these bacteria. However, these methods are time-consuming and labor-intensive. The present study was conducted to evaluate the discriminatory power of two common fingerprinting methods i.e. BOX-polymerase chain reaction (PCR) and enterobacterial repetitive intergenic consensus (ERIC)-PCR for E. faecium clinical isolates.
Methods: Fifty multidrug-resistant E. faecium isolates were isolated from 74 clinical specimens. The isolates were identified by specific 16S rRNA PCR. All isolates were fingerprinted using BOX-PCR and ERIC PCR. The discriminatory power and reproducibility of these two methods were also assessed.
Results: According to the dendrogram with >60% similarity, 17 different genotypes were observed using ERIC PCR. In addition, BOX-PCR produced 22 distinct patterns at a genetic distance percentage of 60%, with sizes ranging from 278 bp to 1450 bp. The discrimination index of BOX-PCR was higher than that of ERIC-PCR.
Conclusion: We concluded that a combination of ERIC-PCR and BOX-PCR may be a quicker and more reliable alternative for the discrimination of E. faecium clinical isolates.

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Background and objectives: Fanconi anemia (FA) is an autosomal recessive disorder that usually manifest in forms of pancytopenia, hyperpigmentation, and skeletal complications. Mutation in the DNA repair regulatory genes is associated with the development of FA. Examination of chromosomal breakages when chromosomes are exposed to cross-linking agents is a common method of FA diagnosis. This study aimed to evaluate the prevalence and characteristics of patients with FA in Mashhad, northeast of Iran.
Methods: In this study, we evaluated 312 suspected FA patients who had been referred to the laboratory of Ghaem Hospital during 2014-2020. The mitomycin C method was used to identify FA-positive subjects.
Results: After the examinations, 84 patients (26.9%) were cytogenetically positive for FA. Of 84 patients, 48 (57.1%) were male and 36 (42.9%) were female. Thumb abnormality was the most common congenital anomaly (43.2%).
Conclusion: Based on the findings, males are more susceptible to FA, and thumb abnormality is the most common congenital anomaly associated with FA. Combination of clinical manifestations and genetic susceptibility in patients may contribute to a more accurate diagnosis.
 

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Background and objectives: Endometriosis is a chronic disease that affects approximately 10% of women of reproductive age. The purpose of this study was to evaluate the effects of 8 weeks of swimming exercise and omega-3 supplementation on the expression of cyclooxygenase-2 (COX-2) and prostaglandin E2 (PGE2) genes and serum levels of reproductive hormones in the endometrial tissue of a rat model of endometriosis.
Methods: In this experimental study, 30 adult Wistar rats were randomly divided into 5 groups of healthy-control, patient-control, patient+exercise, patient+omega-3, and patient+omega-3+exercise groups. After the induction of endometriosis, the rats were subjected to 8 weeks of swimming exercise, 5 days a week as well as 2 ml/kg/body weight of omega-3. Data were analyzed using one-way analysis of variance and Tukey post hoc test at the significance level of 0.05.
Results: The expression of COX-2 and PGE2 as well as serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol levels were significantly higher in the patient group compared with the healthy-control group (p≤0.0001). Exercise and omega-3 supplementation either separately or combined could significantly reduce the expression of both genes and serum levels of LH, FSH, and estradiol (p<0.05). This effect was more profound in the patient+exercise+omega-3 group.
Conclusion: The results of the present study indicate that regulating the expression of COX-2 and PGE2 genes as well as serum levels of reproductive hormones through swimming exercise and omega-3 supplementation can improve endometriosis.

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Background and objectives: Foodborne pathogens can significantly affect the public health and cause medical, social, and economic burden. Listeria monocytogenes, Salmonella ­enterica, and Yersinia enterocolitica are important foodborne pathogens that can cause various diseases. Plant-derived compounds are promising bioactive substances with inhibitory effects against bacteria. Perovskia abrotanoides Kar. is a medical plant with broad therapeutic activities. In the present study, we aimed to investigate the inhibitory effects of P. abrotanoides extracts against some foodborne pathogens.
Methods: Flowering branches of P. abrotanoides were collected in 2018 and 2019 from three different habitats in the eastern Alborz Mountains, Iran. The antimicrobial activity of the extracts was evaluated using the agar well diffusion test. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extracts were determined against L. monocytogenes, S. ­enterica, and Y. enterocolitica. In addition, the antioxidant activity of the extracts was investigated by the DPPH test.
Results: The lowest MIC (200 µg/ml) and MBC (400 µg/ml) values against Y. enterocolitica were related to the ethyl acetate extract of plants collected from habitat 1 in 2019. The lowest MIC (50 µg/ml) and MBC (400 µg/ml) values against L.­­ monocytogenes were related to the dichloromethane extract of plants collected from habitat 1 in 2019. All extracts showed antioxidant properties. Results of one-way ANOVA indicated that the DPPH scavenging activity of extracts from plants collected in 2019 was greater than that of those collected in 2018. In most cases, the methanol and ethyl acetate extracts showed more radical scavenging potential.
Conclusion: It seems that P. abrotanoides is a rich source of antimicrobial and antioxidant compounds with great potential for use in the pharmaceutical and food industries.

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Background: Urinary tract infection (UTI) is one of the most prevalent bacterial diseases worldwide. Escherichia coli is a well-known etiological agent of UTI. The emergence and spread of metallo-beta-lactamase (MBL)-producing E. coli is a serious threat to public health.
This study aimed to investigate the antibiotic resistance pattern and prevalence of MBL-producing E. coli isolated from UTI.
Methods: From January 2020 to June 2021, 1200 urine specimens were collected from patients suspected of having UTI. Antibiotic susceptibility testing was carried out by the disk diffusion method. The prevalence of MBL (blaVIM, blaIMP, blaSPM, and blaNDM) genes was determined by the polymerase chain reaction (PCR) method.
Results: The highest susceptibility was observed against amikacin (96%) and gentamicin (95%). The isolates were mostly resistant against ampicillin (72%) and cephalothin (60%). All carbapenem-resistant isolates were MBL-positive. Based on the results of PCR, 75% of the isolates were blaNDM-positive.
Conclusion: Resistance to some antibiotics, such as ampicillin and cephalothin, was high, and their prescription must be restricted. The prevalence of MBL-producing isolates was not high; however, due to the high level of resistance against other antibiotics, continuous monitoring of MBL-producing isolates is highly essential.

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Background: Cutaneous leishmaniasis (CL) is a prevalent infectious zoonotic disease between humans and animals. Golestan Province is one of the important centers of CL in Iran. The current method for detecting Leishmania parasites in patients is Giemsa-stained direct smear from skin ulcers; however, PCR is a strongly recommended method for epidemiological studies. This study aimed to investigate Leishmania species in negative ulcer smears from patients suspected of having CL referred to Aq Qala Health Center using the multiplex nested PCR method.
Methods: This study was performed on 72 negative ulcer smears from patients suspected of having CL referred to Aq Qala Health Center, Golestan Province, northeastern Iran, from August 2019 to April 2020, using the multiplex nested PCR method to detect Leishmania major and Leishmania tropica species.
Results: Out of 72 samples, 4 (5.55%) samples were positive by multiplex nested PCR. Moreover, all positive samples are related to Leishmania major species.
Conclusion: The multiplex nested PCR method is strongly recommended for detecting Leishmania species in negative ulcer smears from patients suspected of having CL.
 

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