Pasteur Institute of Iran

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  The features of Helicobacter pylori adhesins, their interactions with their host counterparts, regulated and selective gene expressions are amongst the many clever strategies this microorganism undertakes to survive the otherwise sterile gastric milieu. The ingenious crafting of these interactions and the respective host reactions govern, in part, an array of consequences ranging from asymptomatic infections to varying degrees of gastric inflammation, ulcer formation, atrophic, metaplastic and dysplastic changes and ultimately gastric cancer. The most well studied H. pylori adhesins include those which bind host blood group antigens namely BabA and SabA. In this review, I attempt to tell the historical tale of how these moieties and their respective interactions with the host were discovered and characterized. The details of the subsequent applications of these findings in further genotyping studies will be later reviewed to avoid disruption of this crafty tale. J Med Microbiol Infec Dis, 2014, 1 (2): 5 pages.

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Introduction: The most common mode of rabies virus transmission is through a bite wound or contact of broken skin with saliva of a rabid animal. Various other routes of virus transmission include exposure of mucous membranes (i.e. eyes, nose, and mouth) to infected saliva of a rabid animal, aerosol transmission, and corneal transplantation. Laboratory workers during work with rabies virus and veterinarians during examination and surgery of rabid animals may be at risk for exposure to saliva or other infectious fluids splashing into their eyes. The aim of this study was to investigate the possibility of ocular rabies pathogenesis in mice as an animal model. Our results will determine if rabies virus strains challenge virus standard (CVS) and street rabies virus (SRV) are able to infect the central nervous system (CNS) of mice through the ocular route. Methods: This study was performed in two experiments. In experiment 1, different lethal doses of fixed rabies virus strain CVS were made and instilled into both eyes of test mice. In experiment 2, concentrated rabies virus strains CVS and SRV were instilled into both eyes of the test mice. Mice in all groups were kept for 3 months and tested by fluorescent antibody test (FAT) for detection of the presence of viral antigen in brain tissue. Results: Mice with ocular instillation of fixed and street rabies viruses developed no clinical symptoms of rabies and all were healthy and alive during the 3-month observation period. The FAT results were negative in both experiments. Conclusion: Our results suggest that CVS and SRV viruses are not able to infect the CNS of mice via intact conjunctiva and cornea. 

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Introduction: Essential oils are used as flavoring agents in various foods. Layer-by-Layer (LBL) technique is a method in which the material is dipped into a series of different solutions containing oppositely charged polyelectrolytes. The aim of this study is to investigate the effectiveness of a multilayered edible coating with an antimicrobial compound (Lemon and Peppermint) in enhancing the quality and shelf-life of rainbow trout (Oncorhynchus mykiss) during refrigerated storage (4 ± 1˚C) over a period of 16 days. Methods: In this study, multilayered coating was used with two concentrations of Lemon (LEO) and Peppermint (PEO) essential oils (0.5 and 1%). Antibacterial effect of these treatments was evaluated by enumeration of bacteria in special culture media. The control and the coated fish samples were analyzed periodically for pH and microbiological (total viable count, psychrotrophic count, lactic acid bacteria, Enterobacteriaceae, and coliform) characteristics. Results: The results obtained in this study demonstrate that multilayered coating in combination with Lemon and Peppermint essential oils can significantly decrease the number of bacteria and delay the spoilage of the samples (p<0.05). Conclusion: Multilayered edible coating with an antimicrobial compound can properly delay the growth of spoilage microorganisms and prolong the shelf life of meat products.

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Introduction: Helicobacter pylori, as an etiologic cause of peptic ulcers and gastric cancer, should be tested and treated. The true state of infection can only be detected by isolation of gastric biopsies through the invasive method of gastroscopy. However, there are several non-invasive methods for detection of infection, the most common of which is serology. Methods: Here we have evaluated the efficacy of two commonly used commercial IgG-based ELISA kits (Kit-1 and Kit-2) against the endoscopy (biopsy)-based methods of rapid urease test (RUT) and bacterial culture. Our study population included 754 subjects categorized as having: 1) nonulcer dyspepsia (NUD, N=485), 2) peptic ulcer disease (PUD, N=65), and 3) gastric cancer (GC, N=204). Results: The rates of agreement between the results obtained by Kit-1, Kit-2 and both kits with culture/RUT were 62.4% (318 of 500, P=0.0001), 74.4% (183 of 246, P<0.0001), and 81.8% (117 of 143, P<0.0001), respectively. The agreement rate between the two kits, regardless of the results of the culture and RUT, was 83.6% (147 of 176, P<0.0001), leaving 16.4% of the subjects with discrepant results. The sensitivity rate and more drastically the specificity rates (against biopsy-based tests), claimed by Kit-1 (100% and 75%) and Kit-2 (>96% and >99%) were significantly reduced (Kit-1: 94.4% and 40.3%; Kit-2: 86.5% and 71%) for the Iranian population. Conclusion: Our data raises questions regarding the accuracy of commercial IgG-based ELISA kits for the detection of H. pylori infection. Therefore, caution should be practiced when such tests are used as the sole basis of medical decision making.

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A 79-year-old male presented with an ulcerated lesion on the glans penis. Histopathological evaluation of the biopsy from the lesion revealed the presence of Leishmania amastigotes and confirmed the diagnosis of cutaneous leishmaniasis. The patient was treated with several sessions of cryotherapy; the lesion healed and left no scar.

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Introduction: Despite considerable efforts to control AIDS pandemic, it is still one of the significant infectious concerns worldwide. The advance in medical research has led to the development of highly active antiretroviral therapy with a considerable effect to suppress the disease. However, an effective vaccine capable of eradication the HIV pandemic is not available yet. Failure to develop a prophylactic vaccine diverted the efforts to clinical trials of therapeutic vaccines. Methods: Here, we review different approaches to dendritic cell-based HIV therapeutic vaccines. We have summarized the dendritic cell-based trials as HIV therapeutic vaccination, registered in the United States clinical trial database. Results and Conclusion: The strategies applied in the clinical trials were mostly of low success rates; however, by using dendritic cell therapy, they could trigger the host immune response against HIV-1 infections.

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Introduction: Breast cancer is one of the most important causes of mortality in women. Various factors are involved in the development of cancer, including viruses. Toll-like receptors (TLRs) have an essential role in the innate immune system. The present study investigated the relationship between TLR2 rs5743708 polymorphisms and Torque teno virus (TTV) infection with breast cancer. Methods: Blood samples from 80 women with breast cancer and 80 healthy women were collected, and after DNA extraction, the presence of TTV was investigated by a PCR assay and polymorphism in the TLR2 gene (rs5743708) was explored using the PCR-RFLP method. Also, the physical and chemical properties of TLR2 protein in the two wild and mutant forms were analyzed using the ExPASy database. Results: Statistical analysis showed that there was no significant relationship between the age and TTV infection; TTV infection and breast cancer; the grade of cancer, and TTV infection; while there were significant relationships between rs5743708 polymorphisms and breast cancer; GG genotype and increased incidence of cancer; TTV infection and rs5743708 polymorphisms. Also, instability index, aliphatic index, grand average of hydropathicity, and molecular weight of TLR2 protein varied in wild and mutant states. Conclusions: Although there was no significant relationship between TTV infection and breast cancer, the rs5743708 polymorphisms might be involved in TTV infection and breast cancer.

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Introduction: Chronic suppurative otitis media (CSOM) is one of the most common middle ear infections leading to extra and intracranial complications if not diagnosed promptly. Early identification and detection of the etiological agents and antibiotic susceptibility patterns assist in preventing complications. Methods: Two hundred twelve ear swabs were collected using sterile cotton swabs. Direct gram staining was done and then inoculated into blood, MacConkey, and Nutrient agar. Bacterial isolates were identified using conventional methods. According to CLSI guidelines, Antibiotic susceptibility testing was performed by Kirby-Bauer disc diffusion method. Minimum inhibitory concentration (MIC) was performed by the agar dilution method. Extended-spectrum beta-lactamases producing bacteria were detected by the phenotypic confirmatory test and then corroborated by uniplex PCR. Results: Out of 212 samples, 157 samples (74.06%) were culture-positive for bacteria. The isolated bacteria included Pseudomonas aeruginosa (46.24%), Staphylococcus aureus (26.59%), Klebsiella pneumoniae (14.45%), coagulase-negative Staphylococcus aureus (5.20%), Proteus mirabilis (4.05%), Enterococcus faecalis (2.89%), and Escherichia coli (0.58%). The P. aeruginosa isolates showed 96.25% and 95% susceptibility to amikacin and ofloxacin, respectively. All Gram-negative bacilli isolates were 100% sensitivite to imipenem. Ten (30.30%) isolates were ESBL producers with the CTX-M-14 gene detected in most of them. Conclusion: Our study found that P. aeruginosa was the most common isolated pathogen bacteria. Knowledge of CSOM causing bacteria and their susceptibility to antibiotics would help choose an appropriate treatment, thereby preventing antibiotic resistance and complications in these cases.

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Hematopoietic stem cell transplantation (HSCT) severely undermines the recipients' immune status and makes them prone to complications following viral infection. Here, we report a 3-year-old boy with mucopolysaccharidosis type VI who acquired SARS-CoV-2 infection after HSCT. The boy was diagnosed with SARS-CoV-2 during the post-transplant period (19 days after HSCT) when dealing with acute graft versus host disease (GVHD). He was successfully treated with remdesivir and tocilizumab and recovered. Well-timed treatment with tocilizumab might reduce the risk of invasive mechanical ventilation and death in patients with severe COVID-19 pneumonia in the early post HSCT period.

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Introduction: Extra-pulmonary tuberculosis (EPTB) is a significant cause of morbidity, and early diagnosis is critical for improving patient outcomes. Conventional diagnostic methods for EPTB often require improvement, highlighting the need for more rapid and sensitive diagnostic procedures. In this cross-sectional study, we aimed to evaluate the diagnostic usefulness of multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets for detecting Mycobacterium tuberculosis in samples from suspected cases of EPTB. We compared the performance of mPCR with conventional methods, including Ziehl Neelsen (ZN) microscopy, culture in LJ media, and BacT/Alert system. Our study aimed to provide insight into the utility of mPCR and its different targets for diagnosing EPTB in our setting. Methods: We conducted a cross-sectional survey of 250 non-repeat clinical samples from extrapulmonary sites to detect M. tuberculosis. Both conventional diagnostic methods, including ZN microscopy, culture in LJ media, and BacT/Alert system, and molecular methods, including multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets, were performed on the samples. Of the 250 samples, results for all the diagnostic methods were available for 116 samples, which were included in the final analysis. The study population comprised 83 patients with suspected EPTB and 33 controls. Results: Among the 83 samples in the EPTB group, conventional diagnostic methods, including ZN microscopy, LJ culture, and BacT/Alert system, showed low positivity rates of 6.02%, 8.43%, and 15.66%, respectively. In contrast, multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets showed a significantly higher positivity rate of 79.51%. The IS6110 gene was amplified in 79.51% of the samples, while mpb64 was amplified in 49.39%. Conclusion: Our study demonstrates that multiplex PCR (mPCR) using IS6110 and mpb64 as gene targets is a more sensitive diagnostic method for extra-pulmonary tuberculosis (EPTB) than conventional methods. Both IS6110 and mpb64 showed high sensitivity of 100%, but mpb64 was more specific when compared with the gold standard. Our findings suggest that mPCR, particularly with the inclusion of mpb64 as the target gene, may be a valuable tool for the early and accurate diagnosis of EPTB. 

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Introduction: The rising foodborne disease outbreaks poses significant challenges to key objectives in food microbiology. This trend is primarily attributed to global population growth and intensified food production. A thorough microbiological assessment of end products is therefore crucial. Methods: We evaluated the bacterial presence and abundance in various dairy products (sour cream, cottage cheese, buttercream, cream cheese, pasteurized milk, protein-rich milk, and yogurt) sourced from a local supermarket in Bosnia and Herzegovina. Two enumeration methods (pour plating and most probable number) were employed alongside morphological, biochemical, and molecular analyses (Gram staining, oxidase test, catalase test, indole test, lipolytic activity assay, and RT-qPCR). Our focus was on spoilage-causing lactic acid bacteria (LAB), hygiene indicator thermotolerant coliforms (TC), and the foodborne pathogen Salmonella spp.  Results: Six out of seven dairy products harbored high levels of LAB, suggesting potential spoilage, except for cottage cheese. Additionally, both TC and Escherichia coli exceeded acceptable microbial limits, particularly in pasteurized milk. Furthermore, initial tests detected presumptive Salmonella spp. in cream cheese, protein-rich milk, and yogurt. Conclusion: These results highlight the need for stringent sanitary practices during dairy production to extend product shelf-life and prevent premature spoilage from unwanted bacterial presence. Moreover, eliminating pathogen contamination during manufacturing is crucial to mitigate serious food safety risks, including potential food poisoning.

 

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Introduction: Acromegaly is often associated with alterations in carbohydrate metabolism, ranging from impaired glucose tolerance to overt diabetes mellitus (DM). This study aimed to evaluate the serum concentrations of TNF-α and IL-10, along with other biochemical parameters, in patients with acromegaly and concomitant diabetes. Furthermore, we sought to investigate the associations between these parameters. Additionally, this study investigated the prevalence of Cytomegalovirus (CMV) infection and its potential correlation with TNF-α, IL-10, and other biochemical parameters in this patient population. Methods: Serum concentrations of TNF-α and IL-10 were measured in 50 patients with acromegaly and concomitant diabetes and 50 healthy controls using commercially available ELISA kits. CMV DNA was detected in serum samples using a qualitative PCR assay targeting the CMV late antigen gp64 gene. Results: Patients with acromegaly and concomitant diabetes exhibited significantly higher levels of IGF-1, insulin, HOMA-IR, cholesterol, triglycerides, LDL, VLDL, ALT, AST, bone-specific alkaline phosphatase (BALP), TNF-α, and IL-10 compared to the control group (all P<0.05). CMV infection was detected in 1.9% (1/50) of the healthy control group and 23.5% (12/50) of the acromegaly and diabetes group. Within the acromegaly and diabetes group, CMV-positive patients had significantly higher levels of TNF-α and IL-10 compared to CMV-negative patients (both P<0.05). Conclusion: This study demonstrated a significant association between elevated levels of TNF-α and IL-10 and acromegaly with concomitant diabetes. Further research is needed to determine if these cytokines play a causal role in the pathogenesis of these comorbidities. The observed increase in ALT, AST, and BALP levels in patients suggests potential liver and bone involvement in acromegaly with concomitant diabetes. Moreover, a higher prevalence of CMV infection was observed in patients with acromegaly and concomitant diabetes compared to healthy controls, suggesting a potential link between CMV infection and this patient population. Further research is warranted to elucidate the nature of this association and its potential clinical implications.

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Introduction: Green fluorescent protein (GFP) and its variants are pivotal in tracking gene expression across various gene delivery systems. While GFP is typically employed for intracellular reporting, it can be modified to display on cell surfaces for labeling. Previous research indicates GFP might have immunogenic effects, notably enhancing tumor-specific T cell responses. This study explores the immunostimulatory differences between enhanced GFP (EGFP) and the supercharged variant, +36 GFP. Methods: Recombinant EGFP and +36 GFP proteins were generated using an Escherichia coli expression system. Murine bone marrow-derived dendritic cells (BMDCs) were generated using established protocols. Splenocytes were isolated from murine spleens via mechanical disruption and red blood cell lysis. The RAW 264.7 macrophage cell line was cultured in complete DMEM medium. Immune cells were then incubated with varying concentrations of EGFP and +36 GFP, separately, for 48 h. Cytokine levels (IFN-γ, TNF-α, IL-10) were quantified using sandwich ELISA.