Proteases of Acanthamoeba.

The members of genus Acanthamoeba are the etiological agent of uncommon but severe or even fatal opportunistic infections in human beings. The presence of different classes of intracellular and extracellular proteases including serine proteases, cysteine proteases, and metalloproteases has been well documented in environmental and clinical isolates of Acanthamoeba spp. However, the role of the proteolytic enzymes in physiological, biological, and pathological mechanisms of the amoeba remains partially investigated. Some attempts have been conducted using various methods to determine the profile of proteases (number, class, optimal conditions, and activity of the enzymes), and possible pathogenicity mechanism of the proteolytic enzymes (various protein substrate degradation, cytopathic effect on different cell lines). In some cases, it was attempted to correlate intracellular and extracellular protease profile with pathogenicity potential of strains. This review revealed that the protease profile of different strains of Acanthamoeba was extremely complex, therefore, further comprehensive studies with application of a combination of various methods may help to elucidate the role of the enzymes.

Empagliflozin Attenuates Renal and Urinary Markers of Tubular Epithelial Cell Injury in Streptozotocin-induced Diabetic Rats.

Empagliflozin, a SGLT-2 inhibitor, improves diabetic nephropathy through its pleiotropic anti-inflammatory effects. The present study aims to evaluate empagliflozin effects on renal and urinary levels of tubular epithelial cell injury markers in streptozotocin-induced diabetic rats. Empagliflozin at 10 mg/kg (p.o.) was administered for 4 weeks, beginning 8 weeks after induction of diabetes. Renal function as well as markers of renal tubular epithelial cell injury were assessed in kidney tissue homogenates and urine. Empagliflozin was able to ameliorate diabetes induced elevations in serum cystatin C levels. It also alleviated renal KIM-1/NGAL levels and urinary albumin, α-GST, and RBP excretions. In addition to decreasing urinary levels of cell cycle arrest indices i.e. TIMP-2 and IGFBP7, empagliflozin mitigated acetylated NF-κB levels in renal tissues of diabetic rats. As a whole, these findings reveal empagliflozin capability in improving diabetic nephropathy via ameliorating indices of renal inflammation, injury, and cell cycle arrest on streptozotocin-induced diabetic rats.

Laboratory Cross-Contamination of Mycobacterium tuberculosis: A Systematic Review and Meta-analysis.

BACKGROUND: Microbiological cultures are the mainstay of the diagnosis of tuberculosis (TB). False-positive TB results lead to significant unnecessary therapeutic and economic burden and are frequently caused by laboratory cross-contamination. The aim of this meta-analysis was to quantify the prevalence of laboratory cross-contamination. METHODS: Through a systematic review of five electronic databases, we identified studies reporting rates of laboratory cross-contamination, confirmed by molecular techniques in TB cultures. We evaluated the quality of the identified studies using the National Institute of Health (NIH) Quality Assessment Tool for Observational Cohort and Cross-Sectional Studies, and conducted a meta-analysis using standard methodology recommended by the Cochrane Collaboration. RESULTS: Based on 31 eligible studies evaluating 29,839 TB cultures, we found that 2% (95% confidence intervals [CI] 1-2%) of all positive TB cultures represent false-positive results secondary to laboratory cross-contamination. More importantly, we evaluated the rate of laboratory cross-contamination in cases where a single-positive TB culture was available in addition to at least one negative TB culture, and we found a rate of 15% (95% CI 6-33%). Moreover, 9.2% (91/990) of all patients with a preliminary diagnosis of TB had false-positive results and received unnecessary and potentially harmful treatments. CONCLUSIONS: Our results highlight a remarkably high prevalence of false-positive TB results as a result of laboratory cross-contamination, especially in single-positive TB cultures, leading to the administration of unnecessary, harmful treatments. The need for the adoption of strict technical standards for mycobacterial cultures cannot be overstated.

Cytotoxic effect of 2, 5-dimethyl-celecoxib as a structural analog of celecoxib on human colorectal cancer (HT-29) cell line.

Dimethyl-celecoxib (DMC), a close derivative of celecoxib (CXB) with a low COX-2 inhibitory function, exhibits significant anti-neoplastic properties. In this study, we have investigated the effect of CXB and DMC on the human HT-29 cell line. The cellular viability, caspase-3 activity, and VEGF, NF-κB, and COX-2 genes expressions were assessed respectively with MTT, colorimetric, and real-time RT-PCR methods. DMC, a close analogue of CXB, was more potent in inhibiting the growth of cells (IC50: 23.45 µM at 24 hr) than CXB (IC50: 30.41 µM at 24 hr). Both CXB and DMC caused a significant difference in caspase-3 activity compared to the control group. DMC significantly decreased the NF-κB expression. Down-regulation of the COX-2 mRNA expression in the celecoxib-treated group was significant compared with that in the DMC-treated group. Alterations in the mRNA expression of VEGF were not significant between the groups. Owing to the more potent growth inhibitory effects of DMC compared to that of celecoxib, it may be important to conduct research on the anticancer application of this compound, which can reduce the side effects relating to COX2 inhibition.

Frequency of Smear-Negative Tuberculosis in Northwest Iran.

BACKGROUND: Microscopic smear examination is the most common test in tuberculosis (TB) detection. It is, however, not strong enough to identify TB in the majority of afflicted individuals; thus, a significant number of TB patients are smear negative and capable of transmitting the infection. The aim of this study was to evaluate the rate of smear-negative TB in northwest Iran. METHODS: In this cross-sectional study, 329 TB-confirmed patients were evaluated through culture up to March 1, 2015, in northwest Iran. The demographic and clinical features of the smear-negative and smear-positive TB patients were compared. The χ2 test was used to compare the frequency of the variables. All the statistical analyses were conducted using SPSS, version 16 (Chicago, IL, USA). RESULTS: Seventy-five cases were smear negative and 254 were smear positive. Smokers, asthmatics, and extra-pulmonary TB patients were primarily among the smear-negative cases. The rate of mortality was also relatively higher among the smear-negative TB patients. CONCLUSION: Totally, 22.8% of the TB cases in northwest Iran were smear negative, with a relatively higher rate of mortality than those with positive smears. A delay in these patients' return to TB diagnosis and treatment centers increases the chance of transmission to others. This is a very sensitive issue in centers where there is no equipment for TB cultivation. Thus, it is essential to equip centers without TB cultivation facilities and to use appropriate diagnostic techniques in centers with those facilities to help rapidly detect smear-negative cases.

The Association between Diet and Multiple Sclerosis.

BACKGROUND: Multiple sclerosis (MS) is a chronic autoimmune disease of the Central Nervous System (CNS) that affects individuals between the ages of 20 and 40 years, with a higher prevalence among women. Prevalence of this disease has increased significantly in recent decades in different geographical areas. There is evidence to suggest that both genetic and environmental factors play a role in the development of MS. OBJECTIVE: This study aims to investigate the potential relationship between diet and MS in the Azeri population of the East Azerbaijan province of Iran. METHODS: 467 MS patients and 260 non-related healthy individuals under the age of 15 completed a dietary demographic questionnaire. The relationship between food consumption and MS was evaluated using the obtained data. RESULTS: MS patients had a significantly higher consumption of fat, high-fat dairy, fast food, soybean, sausages and kielbasa, pickles, and leftover food (p-value=0.0001), while healthy individuals had a higher consumption of fruit (p-value=0.0001). Consumption of Meat, sweets, and fizzy drinks was also found to be higher in MS patients (p-value<0.05). There was no significant difference in the consumption of vegetables, cakes biscuits, and spices between the two groups (p-value>0.05). CONCLUSION: The results suggest that fruit consumption under the age of 15 may be a protective factoragainst MS, while the consumption of fat, high-fat dairy, fast food, soybean, sausages and kielbasa, pickles, leftover food, meat, sweets, sauce, and fizzy drinks under the age of 15, maybe risk factors for MS.

How Molecular Epidemiology Can Affect Tuberculosis Control in the Middle East Countries: A Systematic Review and Meta-Analysis.

BACKGROUND: Nowadays, due to the incidence of specific strains of Mycobacterium tuberculosis and also an increase in the rate of drug resistant-TB, the mortality rate has been increased by this disease. The identification of common strains in the region, as well as the sources of transmission, is essential to control the disease, which is possible by using molecular epidemiology. OBJECTIVES: In this survey, the studies utilizing the spoligotyping method in Muslim Middle East countries are reviewed to determine their role in the control of TB. METHODS: All studies conducted from 2005 to June 2016 were considered systematically in three electronic databases out of which 23 studies were finally selected. RESULTS: The average rate of clustering was 84% and the rate of recent transmission varied from 21.7% to 92.4%. The incidence of Beijing strains has been found to be rising in the abovementioned countries. In Iran and Saudi Arabia known as immigration and labour-hosting countries, respectively, rapid transmittable and drug-resistant Beijing strains were higher than those in other Muslim Middle East countries. CONCLUSION: Considering the incidence of highly virulent strains, due to the increase in immigration and people infected with HIV, tuberculosis, especially drug-resistant form, careful monitoring is needed.

Interleukin-10 Promoter and the CCR5 Polymorphisms in Iranian Azari Population with Multiple Sclerosis.

BACKGROUND: Changes in the expression of cytokines as the result of the single nucleotide polymorphisms (SNPs), can affect the incidence of multiple sclerosis (MS). OBJECTIVE: To investigate the relationship between the frequencies of interleukin-10 (IL-10)-1082 A/G (rs1800896) and CCR5-delta32 genotypes and susceptibility to MS in the Iranian Azari population. METHODS: IL-10-1082 A/G SNP and the CCR5-delta32 were genotyped in 152 patients suffering from MS and 242 healthy non-relatives by allele specific-PCR and simple PCR methods, respectively. RESULTS: The frequencies of AA (37.6%) and AG (55.9%) genotypes of IL-10-1082 were significantly high in the control (p = 0.021) and MS patients (p = 0.015), respectively, with no statistical difference between these groups. There was no significant difference in the CCR5 gene based on the possession of wild/wild and wild/del32 genotypes between MS patients and the control group. The del32/del32 genotype was not seen in any of the investigated groups. Tobacco (cigarettes and hookahs) consumption was higher among the MS patients (p=0.004), and this has the potential to raise the risk of MS in both the individuals and their family. However, it had no significant relation with the frequency of different genotypes of the IL-10-1082 and the CCR5. CONCLUSION: Our finding conclude on possible role of AA genotype of IL-10 -1082 as a protective factor in MS.

Mixed tuberculosis infections in Northwest of Iran.

Primary Mycobacterium tuberculosis (MTB) infection doesn't provide protection against secondary infection and patients can suffer from multiple strains of M. tuberculosis simultaneously. The aim of this study was to use molecular genotyping to identify cases of mixed infection in Northwest of Iran. One hundred and twenty-one positive culture isolates of M. tuberculosis were prepared from patients consecutively in Northwest of Iran from March 2017 to March 2018 and then microevolution and mix infection were assessed using the mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) method. MIRU-VNTR analysis revealed that nine samples (7.3%) had double alleles in at least one locus, as observed by five cases of microevolution, and four cases (3.3%) of mixed infection. According to this study, mixed infection in Northwest of Iran has significantly decreased compared to 13 years ago (7.1% decreased to 3.3%), and in order to eradicate tuberculosis it is necessary to identify all cases of mixed infection, at least in recurrent cases, in the future.

Why the COVID-19 Is Not Significantly Reduced in Iran?

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was transported from China to Iran via passengers. The virus is transmitted through droplets from coughing, sneezing, talking and contact with infected surfaces. Due to high transmissibility in places such as clinics, medical offices, buses and offices where there are large crowds, it spread rapidly in Iran and caused the mortality of a significant number of people, especially the elderly with underlying disease. Preventing travel and gatherings, as well as applying house quarantine in Apr 2020, reduced COVID-19 somewhat but then due to lack of attention to social distancing, reducing the use of masks by the people and inappropriate decisions by the national committee on combating coronavirus (NCCC), including reducing the working hours of public offices and shops, reducing the staff of government offices, monopoly on the distribution of essential materials and supplies, the presence of overcrowding in clinics of medical centers, hospitals, and private clinics caused the decline of the disease in the country to decrease. Measures can be taken to prevent the spread of the virus as much as possible, including: rapid detection of infected people, their isolation, contact tracing, quarantine of people over 65 and under 15, protection of the elderly, prevention of gatherings, forcing the use of protective equipment in possible gatherings, and electronicizing public services.

Vitamin supplementation as a potential adjunctive therapeutic approach for COVID-19: biological and clinical plausibility.

The recent pandemic, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of Coronavirus Disease 2019 (COVID-19) has introduced itself into the human population in the 21st century after the coronavirus diseases SARS-CoV and Middle East respiratory syndrome (MERS-CoV). Major investigations are underway worldwide in the search for pharmaceutical interventions for COVID-19 and many agents are administered in off-label routes. Several cases are under study to check or restrict clinical manifestations of COVID-19. According to the fact that the efficacy of some micro-nutrients like vitamins is proven to treat or prevent infectious diseases because of their antimicrobial and immunomodulatory activity, the potential role of vitamins in the COVID-19 treatment or prevention must be considered.

IFN-γ and TNF-α Gene Polymorphisms in Multiple Sclerosis Patients in Northwest Iran.

BACKGROUND: Cytokines are polypeptides that play critical roles in immune responses. Gene polymorphisms occurring in the inflammatory cytokines are taking a role in autoimmune diseases, including multiple sclerosis (MS), which may induce inappropriate immune responses. OBJECTIVE: The aim of this study was to investigate the allelic and genotypic frequencies of interferon gamma gene (IFN-γ) at +874A/T locus and tumor necrosis factor (TNF-α) at+308A/G locus in MS patients of Azeri population. METHODS: At first, a questionnaire was prepared for each of 240 healthy, non-relative, and 152 Azeri MS patients before obtaining the blood sample from all subjects. After DNA extraction, the frequency of alleles and genotypes of the IFN-γ and TNF-α genes at +874A/T and -308G/A loci, respectively, were determined by allele-specific PCR method. Finally, the frequencies were compared between control and MS patients by chi-square test (x2-test) and p<0.05 was considered significant. RESULTS: In the IFN-γ +874A/T gene single nucleotide polymorphism (SNP), the most allelic and genotypic frequencies in MS patients were the A allele, 55.26% (p=0.04) and the AT genotype, 52.63% (p=0.048). In healthy individuals, it was 65.42% for the A allele and 45.42% for the AA genotype. For the TNF-α 308 G/A SNP, the highest allelic and genotypic frequencies in MS patients were the G allele with 55.92% (p<0.001) and AG genotype with 61.84%, and in healthy subjects, the allelic and genotypic frequencies were 84.2% and 70.8% for the G allele and GG genotype, respectively. CONCLUSION: Head trauma, the infection with the herpes virus and Mycoplasma pneumonia, frequent colds and high consumption of canned foods provide grounds for MS. The T allele in the IFN-γ gene (+874) and the genotypes of AA and AG at the TNF-α gene (-308) at the position-308 were considered as potential risk factors for MS. Therefore, the polymorphisms in cytokine genes and following changes in their expression levels can be effective in susceptibility to MS.

Vitamin D Receptor Gene Polymorphism and the Risk of Multiple Sclerosis in the Azeri Population of Iran.

BACKGROUND: Multiple sclerosis (MS), a disease of the central nervous system (CNS), is associated with damage to the myelin sheath of neurons. It is demonstrated that vitamin D deficiency plays an important role in the development of the disease. Binding of vitamin D to its specific nuclear receptors is a way to exert its function. OBJECTIVE: Possible correlation between the vitamin D receptor (VDR) gene and MS was evaluated in the Azeri population of Iran. METHODS: Different genotypes of the Bsml site were determined by using the PCR-RFLP method in 148 MS patients and 220 non-relative healthy controls. RESULTS: In MS patients, genotype bb was significantly higher than the healthy controls (p<0.05). Additionally, most subjects of the MS group had been insufficiently exposed to sunlight before the age of 15 (p<0.001). Our findings indicated that the red meat intake in MS patients was significantly higher than the healthy controls (p<0.001). In addition, the healthy controls had appropriate dieting behaviors in comparison to MS patients (excessive intake of some foods) (p=0.0001). CONCLUSION: In conclusion, genotype BB and sufficient exposure to sunlight before the age of 15 were the protective factors against MS. Although, excessive consumption of red meat and inappropriate dieting behaviors were predisposing factors to MS disease.

Risk Factors for COVID-19.

SARS-CoV-2 in bats was transmitted to humans at a live and wet animal market in China through the intermediate host, creating COVID-19. Viral, environmental and host factors play roles in virus infection and disease. The virus has high transmissibility and is rapidly transmitted to people through close contact and droplets from coughing, sneezing and talking loudly, as well as through contact with contaminated objects. As crowding is an environmental risk factor for contamination, its transmission is high among patients and staff in hospital and also in elderly-care centers. It is more common in the elderly, in men, and subjects with diabetes mellitus, hypertension, cardiovascular disease, and malignancy.

Role of Immigration in Tuberculosis Transmission to Iran: A Systematic Review.

BACKGROUND: Today, because of increasing immigration and the prevalence of drug-resistant tuberculosis in Iran, identifying intra-community cases is necessary in the country. It will be possible through the use of molecular epidemiologic methods. In this inquiry, in order to determine the role of immigrants in the transmission of specific strains to Iran, the studies have been examined which had been conducted based on molecular epidemiologic methods among Iranians and non-Iranians people. METHODS: All studies from 1997 to the end of March 2017 were examined in three databases of PubMed, Scopus, and Google Scholar and finally, 16 studies were selected. RESULTS: The common clustering rate between Iranians and non-Iranians was determined to be 19.8, and the intra-community recent transmission rate was from 0% to 49% with average of 18.1%. The rate of multidrug-resistant tuberculosis (MDR-TB) was 12.5%, which was higher among immigrants, especially Afghans, and a significant number of the strains were Beijing. CONCLUSIONS: The studies have shown that migrants, especially Afghans, are more effective in transmitting specific strains of tuberculosis to migratory areas. To control tuberculosis, it is necessary to register of immigrant's health information, while enter to the country, so that, by doing appropriate diagnostic tests, the curing the patients, the transmission of tuberculosis to the country would be prevented.

Tuberculosis and Diabetes Mellitus in Northwest of Iran.

BACKGROUND: Planning for control of tuberculosis would need to screen and identify individuals susceptible to TB. Due to the weakness of immune system in diabetic patients, it is more likely for them to reactivate latent TB infection. Regarding the increasing number of diabetics in the community, in this study efforts have been made to estimate the frequency of individuals who have tuberculosis and diabetes mellitus (TB-DM) simultaneously, as it could help making preventive decisions to reduce TB in this part of Iran. MATERIALS AND METHODS: In this study, 329 cases of confirmed TB patients were divided into two groups of diabetic and non-diabetic, then demographic information and clinical variables have been compared between the two groups. RESULTS: Among the examined subjects, 47 patients (14.29 %) had suffered from diabetes mellitus and tuberculosis. All of the DM patients had pulmonary tuberculosis and 87.23% of them were over the age of 50. CONCLUSION: Majority of DM-TB patients were over 50 years of age and also more than half of them were women. So it seems that for DM women over the age of 50, to detect tuberculosis, screening tests such as PPD may be necessary.