فهرست مطالب ehsan aryan

Tuberculosis (TB), a contagious disease caused by Mycobacterium tuberculosis  (M. tuberculosis), remains a health problem worldwide and this infection has the highest mortality rate among bacterial infections. Current studies suggest that intranasal administration of new TB vaccines could enhance the immunogenicity of M. tuberculosis antigens. Hence, we aim to evaluate the protective efficacy and immunogenicity of HspX/EsxS fusion protein of M. tuberculosis along with ISCOMATRIX and PLUSCOM nano-adjuvants and MPLA through intranasal administration in a mice model.

In the present study, the recombinant fusion protein was expressed in Escherichia coli and purified and used to prepare different nanoparticle formulations in combination with ISCOMATRIX and PLUSCOM nano-adjuvants and MPLA. Mice were intranasally vaccinated with each formulation three times at an interval of 2 weeks. Three weeks after the final vaccination, IFN-γ, IL-4. IL-17, and TGF-β concentrations in the supernatant of cultured splenocytes of vaccinated mice as well as serum titers of IgG1 and IgG2a and sIgA titers in nasal lavage were determined.

According to obtained results, intranasally vaccinated mice with formulations containing ISCOMATRIX and PLUSCOM nano-adjuvants and MPLA could effectively induce IFN-γ and sIgA responses. Moreover, both HspX/EsxS/ISCOMATRIX/MPLA and HspX/EsxS/PLUSCOM/MPLA and their BCG booster formulation could strongly stimulate the immune system and enhance the immunogenicity of M. tuberculosis antigens.

The results demonstrate the potential of HspX/EsxS-fused protein in combination with ISCOMATRIX, PLUSCOM, and MPLA after nasal administration in enhancing the immune response against M. tuberculosis antigens. Both nanoparticles were good adjuvants in order to promote the immunogenicity of TB-fused antigens. So, nasal immunization with these formulations, could induce immune responses and be considered a new TB vaccine or a BCG booster.

Multiple Sclerosis (MS) is a multifactorial disease that causes neurological disability. Human retroviruses may have a role in the etiology of several acute and chronic neurological disorders.

We aimed to investigate the presence of HTLV-I/II and HIV antibodies in the sera of MSpatients to assess the relationship between these infections and MS.

This case-control study was conducted on serum samples of MS patients and individuals admitted to the same hospital with no symptoms of MS as the control group. Samples were examined for HIV and HTLV-I/II antibodies using ELISA kits.

A total of 79 people, including 38 MS patients and 41 healthy individuals, were evaluated. The frequency of HIV antibodies was 0.0% in both the case and control groups. The HTLV-I/II antibodies were detected in 1 patient in the control group andnopatients in the MS group, with no statistically significant difference between the two groups (P: 0.999).

This study could not establish a relationship between the presence of HTLV-I/II or HIV antibodies and MS.

Microorganisms living in the human body play an important role in human health. Human microbiota is known as an essential agent and playing an important role in the biological processes of the human body. The role of the microbiota in the host metabolism, intestinal inflammatory responses and their treatment has been well recognized. In this study we investigated the relationship between intestinal microbiota and schizophrenia.

Using keywords of”Gut microbiota”، “Schizophrenia”، “Fecal transplantation” و “Mental disorders”, we searched data bases of  SCOPUS، PubMed و Web of Science in order to find studies on the effects and relationship of the gut microbiota with occurrence of some diseases, the gut-brain relationship, and the role of fecal transplantation in the treatment of inflammatory bowel diseases and mental illnesses, particularly schizophrenia.

Recent studies have emphasized the importance of the microbial gut-brain axis. The intestinal microbiota has certain impacts in the process of development of mental disorders such as anxiety, stress, depression, and behavioral disorders.

Use of gut microbiota maybe helpful for the patients with schizophrenia for improvement of the cognitive changes. In addition, due to the low prevalence and high rate of complications of this disease, more studies on the role of fecal microbiota transplantation in the patients with schizophrenia are recommended.

Herpes simplex viruses have been implicated as a cause of cardiovascular disease (CVD) due to their widespread distribution and ability to infect human vascular endothelial cells.

The present study aimed to evaluate the link between Herpes simplex infections and cardiovascular disease.

This case-control study involved 236 patients aged 35 - 65, 118 with known cardiovascular disease and 118 controls. Patients’ cardiovascular disease evaluation was based on data from questionnaires, a specialist’s physical examinations, and paraclinical tests. Herpes simplex viruses (HSV)-antibodies (IgM, IgG) were determined using ELISA in serum samples, and the biochemical blood indicators were analyzed to examine their relationship to the level of HSV antibodies. All the data were analyzed using SPSS software version 20.

IgM antibodies against herpes simplex were negative among all 236 patients. The positivity rate of IgG antibodies against herpes simplex was 58.8 and 51.2% in the case and control groups, respectively with no significant difference (P = 0.253). Patients with cardiovascular disease had a 0.587-fold higher positive rate of IgG antibodies than patients in the control group (odds ratio (OR) = 0.587). The mean age of patients with positive IgG antibodies was significantly lower than patients with negative IgG antibodies (P = 0.012). IgG positivity among men and women (P = 0.670) and different job statuses (P = 0.866) were not significantly different.

The positivity rate of IgG antibodies against herpes simplex was not significantly different among the study groups. Although patients with positive IgG antibodies’ mean age was lower than those with negative IgG antibodies, the gender and job status were not different.

Hepatitis Cvirus(HCV) infection is one of the leading causesof morbidity and mortality worldwide. It has been hypothesized that a number of bacteria and viruses might be involved in the pathogenesis of cardiovascular disease. The aim of this study was to define the prevalence of HCV in patients with cardiovascular disease in comparison with a control group.

In this study, 281 individuals including 143 cardiovascular patients and 138 healthy controls were assessed for identification of HCV antibodies. The data collection was done between April 2016 and February 2017. The prevalence of HCV antibodies was determined by the enzyme-linked immunosorbent assay (ELISA) method.

There was no HCV infection in both patients with or without cardiovascular disease. There was a significant direct correlation between cardiovascular diseases and mean level of FPG (Fasting plasma glucose) (p<0.001). Also the Systolic and Diastolic blood pressures were significantly higher in the patients with cardiovascular disease (p<0.001 and p=0.005,respectively).

The results of this study show that no evidence of HCV infection is found among a group of cardiovascular patients in the city of Mashhad.

The world is currently challenging with the COVID-19 pandemic due to the SARS-CoV-2, a new member of coronaviruses which emerged in late December 2019. The rapid transmission of the disease made it a global concern that has attracted worldwide attention. As there have been no promising treatments or specific vaccines yet, the most important key to control the pandemic is an early diagnosis. Accordingly, performing diagnostic tests accelerates case detection and prevents further transmissions. Current available tests such as quantitative real-time polymerase chain reaction have some limitations. Therefore, new strategies should be developed for accurate and rapid detection of COVID-19, a life-threatening disease. Biosensing is one of the novel approaches for the SARS-CoV-2 detection, having the potential for rapid and early diagnosis to control this pandemic. RNA, antigens, and antibodies are the main targets in COVID-19 biosensors. Although there have been limited reported studies of COVID-19 biosensing strategies, this review summarized the recent progress in this field.

The line probe assay (LPA) is one of the most accurate diagnostic tools for detection of different Mycobacterium species. Several commercial kits based on the LPA for detection of Mycobacterium species are currently available. Because of their high cost, especially for underdeveloped and developing countries, and the discrepancy of non-tuberculous mycobacteria (NTM) prevalence across geographic regions, it would be reasonable to consider the development of an in-house LPA. The aim of this study was to develop an LPA to detect and differentiate mycobacterial species and to evaluate the usefulness of PCR-LPA for direct application on clinical samples.

One pair of biotinylated primers and 15 designed DNA oligonucleotide probes were used based on multiple aligned internal transcribed spacer (ITS) sequences. Specific binding of the PCR-amplified products to the probes immobilized on nitrocellulose membrane strips was evaluated by the hybridization method. Experiments were performed three times on separate days to evaluate the assay’s repeatability. The PCR-LPA was evaluated directly on nine clinical samples and their cultivated isolates.

All 15 probes used in this study hybridized specifically to ITS sequences of the corresponding standard species. Results were reproducible for all the strains on different days. Mycobacterium species of the nine clinical specimens and their cultivated isolates were correctly identified by PCR-LPA and confirmed by sequencing.

In this study, we describe a PCR-LPA that is readily applicable in the clinical laboratory. The assay is fast, cost-effective, highly specific, and requires no radioactive materials.

Genital infection caused by Herpes simplex virus (HSV) is one of the most common health problems, worldwide. Several methods such as cell culture, serological and molecular methods have been used to detect this virus. Currently, Real-Time Polymerase Chain Reaction (Real time-PCR) technique is widely used due to its high sensitivity and specificity. Besides, Real time-PCR can be employed in the follow-up of therapeutic effects in HSV-infected person who is being treated with antiretroviral drugs. We conducted a review on traditional and current diagnostic methods with a focus on their limitations in the diagnosis of HSV infection.

Tuberculosis (TB) is the leading cause of death by infectious diseases worldwide, and especially prevalent in developing countries. Several vaccines against TB have been developed, recently. The aim of the present study was to design and construct a cloning vector encoding Mycobacterium tuberculosis (MTB) mpt51 gene. DNA was extracted from MTB H37Rv strain. Gene-specific primers were designed using Gene Runner software and the mpt51 gene was amplified by PCR. The amplified fragment and pcDNA3.1(+) cloning vector were both digested with restriction enzymes, the mpt51 fragment was ligated into the vector, and the Escherichia coli (E. coli) TOP10 strain were transformed by the recombinant plasmid. Positive clones were identified by colony PCR, restriction enzyme digestion, and DNA sequencing. The mpt51 gene was successfully cloned into pcDNA3.1(+). A 6400 bp band for the pcDNA3.1(+)/mpt51 recombinant plasmid and a 926 bp band for mpt51 were observed by colony PCR, and restriction enzyme digestion on agarose gels. The DNA sequence was 100% homologous with the mpt51 fragment of H37Rv in GenBank. In the current study, the mpt51 gene of MTB was correctly cloned into pcDNA3.1(+). The expression of this recombinant vector can be studied in eukaryotic cells. Moreover, it is possible to determine the efficacy of this vector as a DNA vaccine candidate, and to test its protective function compared to BCG in animal models in future.

Tuberculosis caused by Mycobacterium tuberculosis (M. tuberculosis), remains as one of the leading causes of deaths worldwide, with nearly two million death cases annually. BCG (Bacille Calmette-Guerin) continues to be the most widely used vaccine in the world, but the protective immunity differs in different parts of the world. Accordingly, new strategies including DNA vaccines are essentially needed. This study was aimed to design and construct a cloning vector containing mpt64 gene of M. tuberculosis. M. tuberculosis H37Rv was cultured on Lowenstein Jensen medium, and genomic DNA was extracted. The mpt64 gene was amplified by PCR using designed specific primers. After the digestion of mpt64 and pcDNA3.1 (+) by BamHI and EcoRI restriction enzymes, the mpt64 fragment was ligated into the digested vector using T4 DNA ligase enzyme. Then, the recombinant vector was transformed into competent Escherichia coli (E. coli) TOP10 strain. To confirm the colonies of transformed bacteria, antibiotic resistance, colony-PCR, restriction enzyme digestion and DNA sequencing were used. To confirm the clones, colony-PCR using mpt64 specific primers was performed and the fragment of 718 bp was observed by gel electrophoresis. Clones were also verified by restriction enzyme digestion using BamHI and EcoRI restriction enzymes and the 718 bp fragment was observed. Furthermore, results of DNA sequencing showed 100% homology with the mpt64 fragment of H37Rv in GenBank. In this study, the mpt64 fragment was successfully cloned in pcDNA3.1 (+) vector. This construct can be used in future studies as a DNA vaccine in animal models to induce immune system responses. Key words: Mycobacterium tuberculosis, DNA vaccine, cloning vector, mpt64

Today, several vaccines have been developed to prevent infectious diseases. The older first-generation vaccines may have many problems. In this regard, genetic engineering plays an important role using tools such as shuttle vectors to develop recombinant DNA vaccines that usually include plasmid constructed so that can propagate in two different host species. The present study reviews a variety of shuttle vectors, their structures, productions, pathogenicity and more importantly their applications in the production of novel vaccines. A systematic review was performed based on search in international databases with no time limit including Scopus, PubMed and Google Scholar. All databases were searched using the standard (English and Persian) keywords. Relevant articles from 1996 to 2018 were collected from search of international databases including Science Direct, Google Scholar, and PubMed using keywords such as “shuttle vectors”, “recombinant plasmids” and “DNA vaccines”. In this study, a total of 31 full texts were used. A shuttle vector typically contains similar components to replication origins and promoters and can propagate in various hosts. Nowadays, they are used in designing and constructing of new vaccines against infectious diseases including tuberculosis and viral hepatitis. Also, Multi-epitope peptide DNA vaccines are effective against some viruses and they are potentially effective against some bacteria such as Helicobacter pylori. Shuttle vectors as a powerful genetic engineering tool have a high ability to study the mechanisms of pathogenic microorganisms and make new vaccines such as DNA vaccines and multi-epitope vaccines. The hope is that such multi-epitope DNA vaccines might induce immunity against multiple antigenic targets, multiple strain variants, and/or even multiple pathogens. However, the ability of DNA vaccination to co-deliver a series of antibody and/or CD4 T cell epitopes remains largely unexplored.

Tuberculosis (TB) remains a major cause of death around the world. Bacillus Calmette Guérin (BCG) is the only vaccine used in TB prevention that has a protective effect in children, but its effectiveness declines in adults. Design and development of new vaccines is the most effective way against TB.
The aim of this study was to design and construct a DNA vaccine encoding mtb32C and mpt51 fusion genes of Mycobacterium tuberculosis. First, mpt51 fragment was amplified by PCR method. The pcDNA3.1+/mtb32C plasmid was transformed into E. coli JM109 and then extracted. The mpt51 gene and pcDNA3.1+/mtb32C plasmid were both digested with EcoRI and BamHI restriction enzymes followed by ligation of mpt51 fragment into the digested vector. The recombinant plasmid containing mtb32C and mpt51 was subsequently transformed into competent E. coli TOP10 strain. The clones were confirmed by colony-PCR, restriction enzyme digestion and sequencing. Using agarose gel electrophoresis, a 926 bp fragment corresponded to mpt51 was observed. Digestion of the vector pcDNa3.1+/mtb32C and mpt51 gene was confirmed by electrophoresis. Then, the pcDNA3.1+/mtb32C plasmid was extracted. Sequencing results confirmed the accuracy of the desired plasmid. In this study, we constructed a cloning vector encoding Mtb32C/Mpt51 gene of Mycobacterium tuberculosis. The eukaryotic expression of this vector can be confirmed in future studies. It can be considered as a DNA vaccine in animal models later. Successful cloning provides a basis for the development of new DNA vaccines against tuberculosis.

As one of the major causes of hospital and community acquired infections, Methicillin-resistant Staphylococcus aureus (MRSA) requires accurate and timely diagnosis. This study aimed to investigate the prevalence and antibiotic resistance patterns of MRSA in patients hospitalized in the Birjand-based Imam Reza hospital.
In this cross-sectional study, a total of 102 clinical Staphylococcus aureus isolates were evaluated. Staphylococcus aureus isolates were confirmed via conventional microbiological and PCR methods (coa gene). The antimicrobial resistance patterns of the isolates were determined using the Kirby-Bauer disk-diffusion based on CLSI guidelines. Resistance to methicillin in the isolates was confirmed by means of PCR method (mceA gene). Finally, the obtained data was analyzed using SPSS software (version 16).
In this study, 50.9% and 58.8% of Staphylococcus aureus isolates were reported as methicillin-resistant using the Kirby-Bauer disk-diffusion and PCR methods, respectively. The highest antibiotic resistance in MRSA strains was found to penicillin (96.6%), to erythromycin (45%), and to ciprofloxacin (36.6%). In present study, resistance to azithromycin, erythromycin, ciprofloxacin, gentamicin, minocycline, and rifampin in MRSA isolates was significantly greater than Methicillin-sensitive Staphylococcus aureus (P A significant percentage of MRSA isolates in the hospitalized patients was resistant to methicillin, which is confirmed even with a wider range in their genotype.

Autophagy is a physiologic process in which double membrane vesicles engulf damaged proteins and organelles for delivering them to lysosomein order to degrade and recycle them via lysosomal digestion. Beclin1 is one of the basic proteins involved in the initial step of autophagosome formation. In the current study, the effect of exogenous Beclin1 to induce autophagy and the effect of 3MA to inhibit of autophagy was assessed in Huh7.5 cells as an in vitro models of hepatocellular carcinoma. The Recombinant pcDNA-Beclin1was transfected into Huh7.5 cells. Also, the cell treated with 3MA. Next, the autophagy induction and inhibition was conducted via LC3 staining as a main autophagy marker using flow cytometry. The result of this study suggest that the over expression of exogenous Beclin1 in Huh7.5 cells elevated the autophagosome formation as shown by intracellular autophagosomal marker LC3-II staining for about 32.32 % and 3MA decreased it up to2% in compared with control cells in which the stained LC3-II was12.08. Recombinant beclin1 may be used as a potential autophagy inducer agent and 3-methyl-Adenin inhibits autophagy formation in Huh7.5 cell. The staining autophagy formation marker LC3-II with specific antibody is a reliable method to measure autophagy activation via flow cytometry.

Iron oxide nanoparticles are among the most effective tools which can replace current medical techniques for diagnosis and treatment of various diseases. Hepatitis C infection is one of the main health problems in the world, affecting around 3% of the world's population. This infection can develop into liver cirrhosis and liver cancer over the time in 80% of patients.
In this study, the effects of PEG interferon loaded iron oxide nanoparticles on hepatitis C virus infection compared with unloaded nanoparticles was studied in vitro. First, Huh7.5 cells were cultured to replicate the hepatitis C virus. After loading the peg interferon alpha on iron oxide nanoparticles, their effects on the replication of hepatitis C virus was investigated by several methods. The results of this study showed that iron oxide nanoparticles and peg interferon loaded iron oxide nanoparticles were able to reduce the load of hepatitis C virus in infected cell culture, but differences were not statistically significant. These data indicated that hepatitis C viral load was decreased in infected cells after induction of PEG interferon loaded iron oxide nanoparticles, but it needs more research to clarify in animal models or even to examine with other types of bare and drug-loaded nanoparticles in a similar way to our study.

Hepatitis C virus (HCV) is a member of the Flaviviridae family, which causes approximately 500,000 deaths annually. HCV infection treatment is often associated with significant adverse effects. Curcumin is an active ingredient of turmeric which has therapeutic anti-inflammatory effects in many diseases including infectious ones. Although curcumin is not soluble in water, if it is synthesized in the form of nanomicelles, it will be water soluble and can be absorbed in the gastrointestinal tract (GI). In this study, the antiviral effects of curcumin nanomicelles were investigated on the attachment and entry of HCV particles.
The cytotoxicity of curcumin nanomicelles was determined in Huh7.5 cells and their antiviral effects on the attachment and entry of HCV was investigated in a cell culture system.
Curcumin nanomicelles could decrease the viral load in the cell culture supernatants compared to virus control.
According to the results of this research, we determined the antiviral effects of curcumin nanomicelles in the later stages of HCV replication.

The purpose of this study was to clone, express, and purify a novel multidomain fusion protein of Micobacterium tuberculosis (Mtb) in a prokaryotic system.
An hspX/esxS gene construct was synthesized and ligated into a pGH plasmid, E. coli TOP10 cells were transformed, and the vector was purified. The vector containing the construct and pET-21b () plasmid were digested with the same enzymes and the construct was ligated into pET-21b (). The accuracy of cloning was confirmed by colony PCR and sequencing. E. coli BL21 cells were transformed with the pET-21b ()/hspX/esxS expression vector and protein expression was evaluated. Finally, the expressed fusion protein was purified on a Ni-IDA column and verified by SDS-PAGE and western blotting.
The hspX/esxS gene construct was inserted into pET-21b () and recombinant protein expression was induced with IPTG in E. coli BL21 cells. Various concentrations of IPTG were tested to determine the optimum concentration for expression induction. The recombinant protein was expressed in insoluble inclusion bodies. Three molar guanidine HCl was used to solubilize the insoluble protein.
An HspX/EsxS Mtb fusion protein was expressed in E. coli and the recombinant protein was purified. After immunological analysis, the HspX/EsxS fusion protein might be an anti-tuberculosis vaccine candidate in future clinical trial studies.

Novel tuberculosis (TB) vaccines that aim to boost and/or replace Bacillus Calmette-Guerin (BCG) are currently in development. DNA vaccines can stimulate both humoral and cell-mediated immunity in different animal models of TB and is thought to be a promising strategy in the development of new vaccines against TB. The aim of this study was to design and construct a DNA vaccine encoding ag85a and tb10.4 fusion genes of Mycobacterium tuberculosis.
tb10.4 fragment was amplified by PCR and the product was digested with restriction enzymes. Next, it was cloned into the pcDNA3.1 plasmid. The ag85a gene and pcDNA3.1ﻖ.4 plasmid were digested by EcoRI and BamHI restriction enzymes. Constructed vector was sequenced. The molecular analysis was done using bioinformatics software. New chimeric vector containing ag85a-tb10.4 genes were purified. Expression of pcDNA3.1ﻖ.4-ag85a plasmid was confirmed in eukaryotic cells.
Fragments of 297 bp for tb10.4 and 1017 bp for ag85a were observed in agarose gel electrophoresis. Alignment of ag85a-tb10.4 genome sequence with reference genes in GenBank showed exact identities that indicate correction of all cloning procedures. Transfection of eukaryotic cells with pcDNA3.1ﻖ.4-ag85a vector and existence of tb10.4-ag85a fusion gene were both confirmed with RT-PCR.
In this study, tb10.4 and ag85a genes were isolated from Mycobacterium tuberculosis H37Rv strain and cloned into pcDNA3.1. Also, the capability of constructed vector in producing fusion ag85a-tb10.4 protein was confirmed with RT-PCR. pcDNA3.1ﻖ.4-ag85a vector can be used for further studies in future.

Tuberculosis (TB) is the most important infectious disease and is one of the most common causes of death in the world especially in developing countries. TB is caused by infection with Mycobacterium tuberculosis. Designing and construction new vaccines against Mycobacterium tuberculosis are the only effective way to prevent and control the disease. The aim of this study was to design and construct a cloning vector encoding hspX and tb10.4 fusion genes of Mycobacterium tuberculosis.
At first, tb10.4 fragment was amplified by PCR method and it was digested with restriction enzymes and was cloned into the plasmid pcDNA3.1 . Then, hspX fragment was amplified by PCR method and it was digested by HindIII and BamHI restriction enzymes. The recombinant plasmid pcDNA3.1 / tb10.4 also digested with the same enzymes and hspX was subcloned into the recombinant vector. This construct was transformed into the Escherichia coli strain TOP10. The confirming the clones were performed by colony PCR, restriction enzyme digestion and sequencing methods.
PCR and enzymatic digestion products were run on the agarose gel and tb10.4 and hspX genes were observed 291bp and 435bp, respectively. In addition, results of DNA sequencing showed 100% homology with hspX and tb10.4 genes of Mycobacterium tuberculosis H37Rv strain recorded in GenBank.
In this study, hspX and tb10.4 genes of Mycobacterium tuberculosis were cloned into pcDNA3.1 vector correctly. This vector can use as a DNA vaccine to induce immune system responses in animal models in future studies.

Tuberculosis (TB) is a major cause of death worldwide. Finding an effective vaccine against TB is the best way to control it. Several vaccines against this disease have been developed but none are completely protective. The aim of this study was to design and construct a cloning vector containing the Mycobacterium tuberculosis (M. tuberculosis) heat shock protein X (hspX).
First, an hspX fragment was amplified by PCR and cloned into plasmid pcDNA3.1() and recombinant vector was confirmed.
A 435 bp hspX fragment was isolated. The fragment was 100% homologous with hspX of M. tuberculosis strain H37Rv in GenBank.
In this study, the cloning vector pcDNA3.1(), containing a 435-bp hspX fragment of M. tuberculosis, was constructed. This could be used as a DNA vaccine to induce immune responses in animal models in future studies.

Mycobacterium tuberculosis is the causative agent of tuberculosis (TB). Approximately one-third of the world's population is infected with M. tuberculosis. Despite the availability of drug and vaccine, it remains one of the leading causes of death in humans especially in developing countries. Epidemiological studies have indicated that only 10-30% of people exposed to tubercle bacillus are infected with M. tuberculosis, and at least 90% of the infected people finally do not acquire TB. The studies have indicated that the host efficient immune system has essential roles in the control of TB infection such that the highest rate of mortality and morbidity is seen in immunocompromised patients such as people infected with HIV.
M. tuberculosis is an obligatory intracellular bacterium. It enters the body mainly through the respiratory tract and alveolar macrophages combat this pathogen most commonly. In addition to alveolar macrophages, various T-cell subpopulations need to be activated to overcome this bacterium's resistance to the host defense systems. CD4 T cells, through production of several cytokines such as IFN-γ and TNF-α, and CD8 T cells, through cytotoxic activities and induction of apoptosis in infected cells, play critical roles in inducing appropriate immune responses against M. tuberculosis.
Although cell-mediated immunity is the cornerstone of host responses against TB and the recent studies have provided evidence for the importance of humoral and innate immune system in the control of TB, a profound understanding of the immune responses would provide a basis for development of new generations of vaccines and drugs. The present study addresses immune responses involved in M. tuberculosis infection.

Vincent’s angina refers to acute necrotizing ulcerative lesions of the oral mucosal membranes and tonsils.. In this case, we report a 17-year old girl with emotional stress referred with symptoms of Vincent’s angina. Gram staining showed numerous fusospirochetal bacteria. With antibiotic therapy, amoxicillin and cephalexin, no clinical improvement was observed but improvement was achieved by using mouthwash and salt water.. It seems that there is a close relationship between stress and the Vincent angina disease. Moreover, simple diagnostic and therapeutic procedures such as Gram staining and using mouthwash and salt water are beneficial in case of Vincent angina..

Although milk is the valorous source of nutrition, consumption of aflatoxin M1 (AFM1) contaminated milk induces problems to human health. AFM1 is found in milk when dairy animals are fed with AFB1-contaminated feedstuff. AFM1 remains stable during milk pasteurization and storage as well as during the preparation of various dairy products.. The aim of the current study was to evaluate AFM1 contamination in raw milk of Fariman city.. During the summer (July to September, 2012), 45 samples of raw milk were collected randomly from Fariman, Iran. The samples were analyzed with a commercial competitive enzyme-linked immunosorbent assay (ELISA) kit.. AFM1 was detected in all milk samples. The mean concentration of AFM1 was 11.61 ± 0.72 ng/L with a range of 6.3-23.3 ng/L. None of the samples had levels exceeding the maximum tolerance limit (50 ng/L) accepted by the European Union.. AFM1 contamination of milk is not a concern in this city. However, dairy cattle feed samples of various livestocks must be regularly checked for aflatoxin and the storage conditions of feeds should be strictly controlled..

Tuberculosis (TB) is regarded as a health problem worldwide, particularly in developing countries. Mycobacterium tuberculosis (M. tuberculosis) is the cause of this disease. Approximately two billion people worldwide are infected by M. tuberculosis and annually about two million individuals die in consequence. Forty million people are estimated to die because of M. tuberculosis over the next 25 years if the measures for controlling this infection are not extensively developed. In the vaccination field, Bacillus Calmette–Guérin (BCG) is still the most effective vaccine but it shows no efficacy in adult pulmonary patients. One of the other problems regarding TB is its appropriate diagnosis.. In this experimental study, the recombinant cytochrome P450 CYP141 protein of M. tuberculosis was expressed and purified to be used as a vaccine candidate and diagnostic purpose in subsequent investigations.. The optimization of the cytochrome P450 CYP141 protein expression was evaluated in different conditions. Then, this protein was purified with a resin column of nickel–nitrilotriacetic acid and investigated via Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) and Western Blotting.. The highest expression of the cytochrome P450 CYP141 protein was obtained by the addition of 1 mM of isopropyl β-D-1-thiogalactopyranoside (IPTG) to the bacterial culture grown to an optical density at 600 nm (OD600) of 0.6, 16 hours after induction. This protein was subsequently purified with a purification of higher than 80%. The results of Western Blotting indicated that the purified protein was specifically detected.. In this experimental study, for the first time in Iran the expression and purification of this recombinant protein was done successfully. This recombinant protein could be used as a vaccine candidate and diagnostic purpose in subsequent investigations..

The occult hepatitis B infection (OBI) is defined as the presence of hepatitis B virus (HBV) DNA in the sera or in the liver biopsy and the absence of hepatitis B surface antigen (HBsAg) by serological test.. The current study aimed to evaluate the occult HBV infection by polymerase chain reaction (PCR) and determine HBV genotyping among the patients with abnormal alanine transaminase (ALT) in Ahvaz city, Iran..Patients and The sera of 120 patients, 54 (45%) females and 66 (55%) males, with abnormal ALT 40-152 IU were collected. All the patients were negative for HBsAg for more than one year. The patients` sera were tested by PCR using primers specified for the S region of HBV. Then the positive PCR products were sequenced to determine HBV genotyping and phylogenic tree.. Of these 120 subjects, 12 (10%) patients including 6 (5%) males and 6 (5%) females were found positive for HBV DNA by PCR, which indicated the presence of occult HBV infection among these patients. The sequencing results revealed that genotype D was predominant with sub-genotyping D1 among OBI patients.. Occult hepatitis B infection is remarkably prevalent in Ahvaz, Iran, and should be considered as a potential risk factor for the transmission of Hepatitis B Virus throughout the community by the carriers..