Abstract

Background and Objective: The aim of this study was to evaluate chemical composition, antibacterial and antifungal effect and antioxidant property of Salvia officinalis, Mentha piperita and Mentha Longifolia.

Material and Methods: At first, chemical analysis of essential oils was determined using GC/MS. Then the antibacterial and antifungal effect of tested essential oils on L. monocytogenes, S. aureus, S. typhimurium and E. coli and two fungal strains including A. niger and A. flavus were determined using disk diffusion agar and broth microdilution methods.  The antioxidant property of essential oils was evaluated using DPPH assay.

Results: Linalool (14.38%), l. menthone (19.03%) and δ-terpinene (21.78%) were the major components of Salvia officinalis, Mentha piperita and Mentha Longifolia, respectively. all tested essential oils had antibacterial effect on foodborne pathogens, which was comparable with tetracycline’s effect. In addition, all essences had appropriate antioxidant potential compared with BHT.

Conclusion: based on the results, Salvia officinalis, Mentha piperita and Mentha Longifolia can be introduced as appropriate natural preservatives.

Keywords: Salvia officinalis; Mentha piperita; Mentha Longifolia, Antibacterial Agents.

Abstract

Background and Objective: Ophthalmic pterygium is a potentially vision-threatening lesion of unknown etiology that often extends on the corneal surface and has a worldwide distribution. Despite various studies, the pathogenesis of pterygium remains unclear and the involvement of human papillomavirus is controversial. We aimed to investigate the involvement of papillomavirus in pterygium formation.

Material and Methods: This case-control study was conducted on 50 tissue specimens of pterygium from the patients who had pterygium surgery as the case group and 10 conjunctival biopsy specimens of individuals without pterygium including the patients with  cataract surgery, as controls. The evidence of papillomavirus infection was tested by polymerase chain reaction (PCR).

Results: All samples, case and control, were not positive for papillomavirus. Both groups were positive for beta-globulin gene used to check the quality of extracted DNA.

Conclusion: In this study, due to the absence of papillomavirus in the context of Pterygium it seems that other factors are involved in causing the disease.

Keywords: Pterygium; Human Papilloma Virus; PCR.

Abstract

       Background and Objective: Various cellular factors affect the process of HIV activity. One of these cellular factors are structures known as microRN that are expected to be involved in controlling HIV replication and infectivity. The expression of one or a set of them may represent the patient's clinical conditions. In this study,  the expression of miR-29a and miR-29b involved in regulating viral genes’ expression was evaluated in three HIV-positive groups and a healthy control group. Later,  the expression level of these microRNAs  was compared between the cases and controls.

      Methods: Total RNA extraction was performed on the collected samples using RNx-plus kit and then the microRNA expression levels were evaluated using Relative Real-time PCR. The obtained data was entered into SPSS 22 and Graphpad softwares and analyzed using Kruskal-Wallis and Man-Whitney tests. P-value of less than 0.05 was considered as statistical significance level.

     Results: The expression level of miR-29a  was reduced in patients under treatment and drug-resistant patients ( P ≤ 0.05)  . All three HIV-positive groups including people without drug treatment, patients under treatment and drug-resistant patients showed reduced miR-29b expression level compared to  control group (P ≤ 0.05).

     Conclusion: the decreased expression of miR-29a and miR-29b in patients under treatment and drug-resistant patients indicates an  increased viral replication and reduced CD4 cell count. It may be possible to predict the progression of the disease by miRNA measurement or control viral replication using these mir-RNAs that requires further studies.

        Keywords: HIV, expression, mir-29a, mir-29b.

Abstract

      Background and objective: Hepatitis B virus (HBV) is a DNA virus with high tendency toward hepatic tissue. There are currently about 3 million HBV-infected people and 350 to 400 million chronic carriers of this virus in the world. X protein plays a role in the over-expression of oncogenes, carcinogenicity of liver cells and overlaps with the basal core promoter of the virus. Mutations at specific nucleotides of this region increase viral replication and liver disease progression. The aim of this study was to investigate the frequency of mutations at nucleotides 1762, 1764 and 1766 of HBV X gene in patients with chronic hepatitis B and hepatitis B-related cirrhosis.

      Methods: In this study, 102 patients including 68 chronic hepatitis patients and 34 patients with hepatitis B-related cirrhosis were enrolled. After DNA extraction, HBV X gene was amplified and sequenced using Semi Nested-PCR. Obtained gene sequences were compared with the standard sequence of HBV virus X gene available in the gene bank (Okamoto AB033559). Then, the mutations in the gene X of HBV were identified.

      Results: Comparison of the standard sequence with sequences obtained from patients showed the presence of A1762T / G1764A mutation in 12 chronic (17.64%) and 13 cirrhotic (38.23%) patients. Also, C1766G / G1764T mutations were found in 8.23% of chronic patients and 17.64% of cirrhotic patients.

      Conclusion: A1762T / G1764A mutations in the overlapping region of the basal core promoter with gene X C-terminal may lead to liver disease progression from chronic hepatitis to cirrhosis, by changing the amino acid sequence of the X protein.

    

ABSTRACT

          Background and Objective: Several studies have

shown the antimicrobial activity of lactobacilli against Salmonella enterica (serotype typhimurium). The aim of this study was to evaluate the inhibitory potential of metabolites produced by probiotic culture of Lactobacillus casei against S. typhimurium and its impact on S. typhimurium motility and biofilm formation.

         Methods: In order to evaluate the impact of the metabolites, L. casei cell-free culture supernatant (CFCS) was collected by centrifugation of L. casei secondary cultures. Effectiveness of the CFCS against Salmonella was evaluated by the well-diffusion method. In addition, in vitro effect of this treatment on motility in Swarm agar and biofilm formation by the bacteria was investigated.

         Results: Inhibition zone diameters of S. typhimurium were 0.83 and 12.1 mm at concentrations of 50 and 100 μl of Lactobacillus CFCS against the log4 of S. typhimurium, respectively. Moreover, CFCS treatment inhibited the motility and biofilm formation by Salmonella. Concentrations of 5% and 10% were determined as the minimum inhibitory concentrations for motility and biofilm formation by S. typhimurium. Furthermore, effectiveness of the CFCS against Salmonella was dose-dependent (P<0.05).

         Conclusion: L. casei CFCS is able to inhibit the growth, motility and biofilm formation in S. typhimurium.

         Keywords: Anti-Bacterial, Lactobacillus Casei Metabolites, Phenotypic Characteristics of S. typhimurium

ABSTRACT

        Background and Objective: In this study, we compared the susceptibility of Salmonella typhimurium phoP mutant and its parent to stress conditions that the pathogen may encounter in a host.

        Methods: For this purpose, we used the phoP deletion mutant constructed in our previous study. In order to test the in vitro susceptibility of the mutant to stress factors, the effect of acidic pH, heat, bile salts and polymyxin on growth of the mutant was examined. Then, minimum inhibitory concentration and minimum bactericidal concentration of bile salts and polymyxin were determined. Salmonella typhimurium 14028 was used as the parent strain.

         Results: The mutant was highly susceptible to bile salts and polymyxin in comparison with the parent strain, but no difference was observed in their susceptibility to acid and heat.

          Conclusion: This study confirms the role of the phoP in resistance of Salmonella to polymyxin and bile salts. Mutation in the phoP leads to susceptibility of the mutant to bile and cationic antimicrobial peptides.

         Keywords: In vitro, Mutant, Salmonella typhimurium, Disease Susceptibility.

ABSTRACT
            Background and Objectives: Diagnosis of hepatitis E virus (HEV) infection could be missed in some cases if serological tests are used solely. Molecular characterization of HEV is essential for diagnosis of acute and chronic HEV infections, and evaluating the chronic HEV infection status in immunocompromised patients. The aim of this study was to prepare a suitable HEV positive control, determine the limit of detection (LOD) of HEV RNA for a specific molecular test, and evaluate the efficiency and precision of the test.
           Methods: Genomic region of HEV NCBI reference sequence was constructed. LOD, intra-assay precision, and inter-assay precision were calculated to evaluate the efficiency and precision of the test. Then, tenfold serial dilutions of the HEV positive control were prepared. Real time PCR was performed three times for each dilution. Mean, standard deviation, and coefficient of variation of cycle thresholds obtained in three independent and simultaneous tests were calculated, and the results were analyzed.
          Results: The LOD of this test was determined as 1.4×104 copy/ml or 42 copy/reaction or 14 copy/µl. Intra-assay precision and inter-assay precision for all assays were lower than 2.5% and 10%, respectively.
          Conclusion: We propose that the real time PCR assay targeting the ORF2/3 overlapping conserved region is suitable for detection of a wide range of different HEV genotypes found in acute and chronic HEV infections. However, the precision of the test should be improved for detecting HEV RNA lower than 103 copy/ml.
          Keywords: Hepatitis E virus, Limit of Detection, Real Time PCR.

ABSTRACT
        Background and Objectives: Human cytomegalovirus (HCMV) is the most common viral cause of morbidity and mortality in immunocompromised patients. The aim of this study was to evaluate the frequency of active CMV infection in hemodialysis patients in Gorgan, Iran.
        Methods: Plasma samples were obtained from 149 hemodialysis patients at Hemodialysis Unit of Panje-Azar Medical Centre in Gorgan, Iran. Presence of CMV-DNA in plasma samples was evaluated by polymerase chain reaction (PCR) using specific primers for highly conserved regions of major capsid protein gene of HCMV. In addition, level of CMV-IgM antibody was measured by serological testing. Demographic information and past medical history of patients were also recorded. Data was analyzed by SPSS software (version 18).
       Results: Total prevalence of CMV infection was 6.7% (10/149) among the patients receiving hemodialysis. CMV-DNA and anti-CMV IgM antibody were detected in 2.68% and 4.69%, of the samples, respectively. One case was found positive for both CMV-DNA and anti-CMV IgM antibody. CMV infection did not have any correlation with gender, age, ethnicity, duration of hemodialysis, and history of blood transfusion.
        Conclusion: A notable proportion of hemodialysis patients in Gorgan have active CMV infection. Accurate detection of these individuals is important for preventing infection spread, especially in immunocompromised individuals. Simultaneous diagnosis of CMV infection using serological testing and PCR assay could help reduce the risk of infection spread.
          Keywords: HCMV, Hemodialysis, PCR, Iran.

ABSTRACT
          Background and Objectives: C-C chemokine receptor type 5 (CCR5) is a chemokine receptor expressed at high levels on the surface of T-cells. A 32-bp deletion in the coding region of the CCR5 (CCR5Δ32) leads to production of an incomplete protein that is not expressed on the cell surface. CCR5Δ32 may be involved in development of autoimmune disease, such as systemic lupus erythematosus. We investigated frequency of the CCR5Δ32 polymorphism in SLE patients and healthy controls, and evaluated the relationship between the CCR5Δ32 polymorphism and susceptibility to SLE in Golestan Province, Iran.
          Methods: Whole blood samples were taken from 80 SLE patients admitted to Shahid Sayyad Shirazi hospital and 80 healthy controls (from a blood bank) in the Golestan Province, in 2016. Baseline clinical and laboratorial characteristics were evaluated regarding the CCR5Δ32 genotypes. The CCR5Δ32 polymorphism was determined from genomic DNA by polymerase chain reaction.
          Result: Genotype frequencies of both groups were in the Hardy-Weinberg equilibrium. The frequencies of the CCR5 and the CCR5Δ32 alleles were 98.13% and 1.88% among the patients, and 98.75% and 1.25% among the controls, respectively. Homozygote CCR5Δ32 was not observed in the subjects. The frequency of heterozygous Δ32 was 3.8% and 2.5% among the SLE patients and controls, respectively (P-value>0.05). There was no significant association between the CCR5 status and clinical signs of SLE (P>0.05).
          Conclusion: Our data suggest that the CCR5Δ32 polymorphism has no correlation with SLE in our study population. In addition, the frequency of the Δ32 polymorphism in SLE patients and controls does not follow the Hardy-Weinberg equilibrium
          Keywords: CCR5, Homozygote CCR5Δ32, Heterozygote CCR5Δ32, CCR5Δ32 allele, SLE.

ABSTRACT
          Background and Objectives: Nonviral carriers including those based on synthetic cationic lipids, offer several advantages over the viral counterparts. These carriers are able to form complexes with nucleic acids and deliver genes into the cells via the cellular endocytosis pathway, without significant toxicity. The level of transgenes expression depends on some experimental variables including cell type and density, Lipofectamine and DNA concentrations and Lipofectamine-DNA complexing time. The main objective of this study was to optimize transfection of SW480 colon cancer cells with Lipofectamine 2000.
          Methods: In this study, SW480 cells were transfected with plasmid containing green fluorescent protein reporter gene using Lipofectamine 2000. Green fluorescent protein expression was studied under a reverse fluorescence microscope and the results were analyzed with the ImageJ software. Effect of different quantities of plasmid DNA and different Lipofectamine 2000 volumes on cell transfection efficiency was evaluated.
          Results: The optimal volume of Lipofectamine and quantity of plasmid was 2 µl and 1µg, respectively, which showed 59% efficiency for the transfection of SW480 cells at 24 hours post-transfection.
          Conclusion: This study shows that Lipofectamine 2000 is an efficient reagent for the delivery of genes into SW480 cells. According to the results, the quantity of DNA per transfection and reagent concentrations are essential factors for a successful transfection.
          Keywords: Optimization; pEGFP-NI; Lipofectamine; SW480.

ABSTRACT
            Background and objectives: Coronaviruses are the main causes of respiratory tract infections in humans. They are also the second leading cause of common cold after rhinoviruses, and can lead to otitis media and asthma. The aim of this study was to investigate the molecular detection of coronaviruses in clinical samples of patients with flu-like symptoms.
            Methods: Specimens were taken from 297 patients with flu-like symptoms who were referred to the influenza laboratory of Golestan University of Medical Sciences during 2012-2014. RNA was extracted from the specimens using an RNA extraction kit. Accordingly, RNA was used for cDNA synthesis and GAPDH was used as the internal control. Synthesized cDNA was investigated for presence of human coronaviruses genome with real-time polymerase chain reaction using specific primers. Data were analyzed by SPSS 16.0 software. 
            Results: The coronavirus genome was not detected in the specimens of patients with flu-like symptoms.
            Conclusion: Genome of human coronaviruses is absent in samples from patients with upper respiratory tract infections and influenza-like symptoms, which may indicate the low prevalence of the virus in the Golestan Province, Iran.
            KEYWORDS: Human coronaviruses, Upper respiratory tract infection, Golestan Province.

ABSTRACT
             Background and objectives: Utilization of essential oils instead of chemical preservatives has received significant attention in recent years. The present study aims to evaluate chemical composition and antibacterial and antioxidant properties of essential oils of Zataria multiflora, Artemisia deracunculus and Mentha piperita.
             Methods: Chemical profile of the essential oils was analyzed by gas chromatography/mass spectrometry. The microwell dilution and agar disk diffusion methods were used to evaluate the antibacterial properties of the essential oils. Total phenolic content, β-carotene-linoleic acid bleaching test and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays were carried out to determine the antioxidant properties.
             Results: Menthol (39.18%) and mentone (21.64%) were the main components of the essential oil of M. piperita, while estragol (34.75%) and limonene (15.72%) were the major components of the essential oil of A. dracunculus. The main components of the essential oil of Z. multiflora were carvacrol (36.81%) and thymol (33.04%). The essential oils of M. piperita and Z. multiflora showed greater antimicrobial effects. Moreover, Z. multiflora showed the greatest antioxidant activity among the essential oils. The total phenolic content of Z. multiflora was 228.14±0.45 mg gallic acid equivalent/g.
             Conclusion: Given their favorable antioxidant and antimicrobial properties, the essential oils of Z. multiflora, A. deracunculus and M. piperita can be used as natural food preservatives.
             Keywords: Zataria multiflora, Artemisia deracunculus, Mentha piperita, antibacterial effect, antioxidant effect.   

ABSTRACT
            Background and objectives: Pterygium is a non-cancerous growth of conjunctival tissue that can extend onto the corneal surface. The presence of some oncogenic viruses in pterygium and the neoplastic nature of these lesions led us to the postulated involvement of the viruses in the etiology of pterygium. Given the association of human herpesvirus 6 (HHV-6) with ocular diseases, we aimed to investigate presence of this virus in pterygium.
            Methods: Fifty tissue specimens were collected from patients with pterygium who underwent pterygium surgery between February 2013 and May 2015. The specimens were tested by real-time PCR using Maxima SYBR Green/ROX qPCR Master Mix (2X) kit. Demographic and clinical data were collected and analyzed using SPSS software (version 18).
            Results: Six (12 %) specimens were positive for HHV-6 DNA. There was no statistically significant correlation between pterygium and presence of HHV-6.
            Conclusion: Based on the results, a direct association between HHV-6 and development of pterygium seems less probable, which suggests that other etiologic agents must be involved in the multistep process of the disease.
            Keywords: Human Herpesvirus 6; pterygium; Real-time PCR.

ABSTRACT
            Background and Objectives: Pterygium is a common ocular surface lesion that manifest as wing-shaped, benign conjunctival growth, which can extend onto the corneal surface. Presence of some oncogenic viruses in pterygium and the neoplastic nature of the lesion led us to the postulated involvement of the viruses in the etiology of pterygia. The aim of this study was to evaluate prevalence and possible role of human cytomegalovirus (HCMV) in the formation of pterygia.
            Methods: Fifty pterygium specimens and 10 normal conjunctival biopsy specimens (controls) were investigated by polymerase chain reaction using primers specific for the highly conserved regions of major capsid protein gene of HCMV. Data were analyzed using SPSS statistical software (IBM SPSS Statistics 18; IBM Corporation, USA) at significance level of 0.05.
            Results: The HCMV DNA was detected in seven (14%) patients with pterygium but in none of the control subjects. All subjects were β-globin positive.
            Conclusion: Given the results, direct involvement of HCMV in the development of pterygium seems less probable, thus suggesting that other agents might be involved in the multistep process of the disease.
            Keywords: Human Cytomegalovirus, Pterygium, Polymerase Chain Reaction.

ABSTRACT
              Background and Objectives: Red blood cell (RBC) transfusion is necessary for the prevention and treatment of a variety of life-threatening injuries and diseases. However, viral contamination of these products is a great threat to recipients. Screening donors for GB virus C by nucleic acid testing is not routinely implemented worldwide. The aim of the present study was to evaluate prevalence of GBV-C RNA in whole blood/red cell components.
              Methods: In this cross sectional pilot study, we collected 153 units of packed RBCs from blood banks of two public hospitals in Gorgan (northeast of Iran), between October and November 2014. The samples were screened for the presence of GBV-C RNA in plasma by nested RT-PCR using specific primers targeting highly conserved regions of 5' UTR of GBV-C. Data were analyzed using SPSS software (version 18).
              Results: Overall, 48 (31.37%) whole blood or red cell components were positive for GBV-C viremia. The GBV-C RNA was detected in 31/88 citrate phosphate dextrose-adenine 1 (CPDA1) RBC, 16/50 washed RBC and 1/13 reduced-leukocyte RBC. However, whole blood CPDA1 was negative for GBV-C viremia. Direct sequencing of PCR products confirmed GBV-C contamination.
              Conclusions: Transmission of GBV-C infection was observed in blood products. Thus, efforts should be made to develop new strategies for assuring blood transfusion safety.
              Keywords: Molecular testing, Epidemiology, Transfusion-transmissible infections, GB Virus C.

Background and objectives: Intestinal parasitic infections are a major public health problem worldwide, especially in developing countries. It is estimated that around 3.5 billion people are infected with intestinal parasites. Human intestinal parasites (HIP) are clinically important due to broad epidemiological distribution, reinfection and drug resistance. In the last decades, bioactive compounds from herbs were used against a wide variety of microorganisms including parasites. We aimed to perform a systematic review on studies on the effects of medicinal herbs on HIPs in Iran.
 
      Methods: Relevant scientific publications until April, 2015 were extracted from five English databases (PubMed, Google Scholar, Ebsco, Science Direct and Scopus) and four Persian databases (Magiran, Irandoc, IranMedex and the Scientific Information Database).
 
      Results: A total of 18 papers and two dissertations met the inclusion criteria. Overall, 22 different plant extracts were used against Giardia lamblia, Entamoeba histolytica, Cryptosporidium and Hymenolepis nana. Based on the results, the extracts could exert time- and dose-dependent inhibitory effects against the tested HIPs. Five plants types including Allium, Chenopodium botrys, Carum copticum, F. asafoetida and Artemisia annua were able to completely inhibit the tested parasites, while Thymus vulgaris and A. paradoxum showed the lowest inhibitory effect (7%).
 
      Conclusions: Given the findings, it is recommended to conduct in vivo studies on medicinal herbs with favorable in vitro effects against HIPs.