Differential Expression of miR-146 and miR-155 in Active and Latent Tuberculosis Infection.

Background: Tuberculosis (TB) is one of the leading causes of death worldwide. Besides, one-third of the world population is infected with Mycobacterium tuberculosis (MTB) while staying clinically asymptomatic; the situation is called latent TB infection (LTBI). MiR-21, miR-31, miR-146a, and miR-155 play an important role in many immune and inflammatory pathways. In the present study the expression levels of MiR-21, miR-31, miR-146a, and miR-155 in peripheral blood mononuclear cells (PBMCs) from patients with active TB, latently infected individuals (LTBI), and healthy controls (HC) were investigated. Participants were recruited at the Bouali Hospital, Zahedan University of Medical Sciences, Zahedan, Iran from 2010 to 2011. Methods: PBMCs were stimulated with PPD before RNA extraction. TaqMan RT-qPCR assay was used to analyze the expression levels of miRNAs. Results: The results indicated no significant differences in the expression of miR-21 and miR-31 between different groups; however, in patients with active TB, the expression of miR-21 (P=0.03) and miR-31 (P=0.04) were significantly increased after stimulation with PPD compared to the unstimulated condition. The expression of miR-146 in response to PPD in both LTBI (P=0.02) and TB (P=0.03) groups compared to the HC group was increased. No significant differences were found in the expression level of miR-155 in response to PPD between LTBI and HC groups. However, the fold change was significantly higher in the TB group in comparison with the HC (P=0.03) and LTBI (P=0.05) groups. Conclusion: The results confirm the main role of miR-146 and miR-155 in TB infection and suggest a role for miR-146 and miR-155 as infection and activation markers in tuberculosis infection, respectively.

Diagnostic Accuracy of Salivary Biomarkers including Lactate Dehydrogenase and Hemoglobin A1c for Screening Chronic Periodontitis.

Aims: Periodontitis is one of the most common chronic bacterial infections in humans involving the tooth-supporting tissue. The present study aimed to evaluate and compare salivary biomarkers, including lactate dehydrogenase (LDH) and hemoglobin A1c (HbA1c), between patients with severe chronic periodontitis and healthy individuals. Methods: This study was performed on 29 patients with severe chronic periodontitis and 30 healthy individuals at Zahedan University of Medical Sciences, Zahedan, Iran, in 2021. Salivary samples were taken, and clinical parameters, including the clinical attachment loss (CAL) and probing pocket depth (PPD), were measured. Besides, the levels of LDH and HbA1c were measured using ELISA kits. The sensitivity, specificity, and positive and negative predictive values of HbA1c and LDH were examined for chronic periodontitis diagnosis. Results: Based on the present results, the levels of LDH and HbA1C did not show adequate sensitivity or specificity for screening chronic periodontitis. Conclusion: According to the present findings, salivary biomarkers, including LDH and HbA1c, cannot be used with certainty for screening chronic periodontitis.

Prevalence of camel babesiosis in southeast of Iran.

Babesiosis is a globally distributed zoonotic parasitic disease in a broad range of vertebrates with great importance in the veterinary field. The standard diagnostic test for Babesiosis in animals is microscopic identification of the parasite in a venous blood smear stained with Giemsa combined with assessment of clinical manifestations throughout the acute phase of the disease. The present study was planned to determine the presence of Babesia species in camels from the southeastern regions of Iran. A total of 140 blood samples of camels were randomly collected in four selected cities including Qaen, Nehbandan, Iranshahr, and Zahedan from March to August 2019. Blood smears of each case were also examined by the Giemsa staining method and extracted DNA samples were subjected to internal transcribed spacers (ITS1) polymerase chain reaction (PCR) amplification. The prevalence rates using microscopically and molecular examinations were 10% and 19.28%, respectively. The prevalence rates significantly vary between the selected regions (p = 0.003). PCR technique showed higher sensitivity than microscopy. We found that all infected camels were positive for Babesia caballi. The rate of infection with Babesia among the camel in Zahedan is remarkable. Early diagnosis and early treatment can prevent further spread of the disease in this area.

Investigation of LAMP Technique in Diagnosis Type of Plasmodium Species in Anopheles Mosquitoes :A Fast and Practical Technique to Detect Malaria Pathogens in the Field.

BACKGROUND: Malaria is one of the main parasitic diseases and a major health issue in some countries. This study aims to determine the rate and type of infections of Anopheles mosquitoes with malaria parasites using the molecular LAMP method in the Southeastern Iran. METHODS: In this study, 400 Anopheles mosquitoes were collected by the Zahedan Medical Insecticide Center in Nikshahr City, a high-risk area of malaria transmission in Sistan-Baluchestan Province. The mosquitoes were caught manually (by hand) in domestic (humans and animals), natural, and artificial outdoor places (Shelter pits). After DNA extraction, the LAMP method was used, which was compared with Multiplex Nested-PCR as a standard method. RESULTS: Out of 400 samples collected from Nikshahr City, 6 samples (1.5%) were infected with Plasmodium vivax. No Plasmodium falciparum or a mix (Plasmodium vivax and Plasmodium falciparum) was detected in this study. CONCLUSIONS: The results of this study indicate that in places with transmission of both species, i.e. Plasmodium vivax and Plasmodium falciparum, detection of malaria parasites by the LAMP method could be very useful in spotting infections in the field. Thus, molecular epidemiological studies could be conducted annually to monitor malaria in endemic regions. The results of this research show that contamination with mosquito malaria vectors is increasing in Nikshahr City, and it seems that more studies will be required to eliminate malaria until 2026.

Salivary ß2-microglobulin levels in patients with erosive oral lichen planus and squamous cell carcinoma.

OBJECTIVES: ß2 microglobulin, as a biomarker, is used for the diagnosis of oral malignant and pre-malignant lesions. The components of the microglobulin system can directly or indirectly help grow and develop tumors. The present study aims to compare beta-2 microglobulin levels in patients with lichen planus of the esophagus, oral squamous cell carcinoma, and healthy individuals. Further, it evaluated the salivary ß2-microglobulin level in malignant and pre-malignant lesions. Oral lichen planus (OLP) is a chronic skin-mucus disorder. Of the total 75 patients referred to Oral Medicine at Dentistry School of Zahedan University of Medical Sciences, 25 were healthy and 25 had oral lichen planus (OLP) and the rest had squamous cell carcinoma (SCC). To collect the saliva samples, unstimulated spitting was used. They were collected between 9 and 12 a.m. Salivary beta2 microglobulin was recorded based on the factory instructions by ELISA optical density method with 450 nm wavelength for each sample. The data were analyzed using descriptive, Kruskal-Wallis and Mann-Whitney and Pearson's correlation coefficient (SPSS 21). RESULTS: The salivary ß2 microglobulin level in patients with squamous cell carcinoma (SCC) and oral lichen plan (OLP) is significantly higher than that in healthy group. Thus, this index is used for assessing early malignant transformation and oral pre-malignant lesion.

Immunogenicity and protection effects of cationic liposome containing imiquimod adjuvant on leishmaniasis in BALB/c mice.

OBJECTIVES: Protection against leishmaniasis, in the murine model, is dependent on developing a potent CD4+ mediated Th1 type response. Liposomes can be applied as immunoadjuvants to stimulate immune responses to different antigens. In the present study, it was investigated whether DOTAP liposomes having SLA and imiquimod adjuvant, can induce a Th1 response and protect against Leishmania major challenge in BALB/c mice. MATERIALS AND METHODS: Liposomes were provided applying the lipid film procedure. BALB/C mice were subcutaneously immunized, three times with 2-week intervals, with various formulations. Assessment of lesion development and parasite burden in the foot and spleen after challenge with L. major, assessment of Th1 cytokine (IFN-γ), and titration of IgG isotypes assessed the type of generated immune reaction and the protection extent. RESULTS: The mice immunized with Liposome DOTAP+imiquimod+SLA showed smaller footpad swelling which was meaningfully different (P<0.05) compared with other groups. The highest level of IgG2a was observed with Lip DOTAP+imiquimod+SLA more than the control (P<0.001). Mice immunized with Lip DOTAP+SLA+imiquimod demonstrated the least number of live parasites in the footpad and spleen. Cytokine assay showed that the greatest IFN- γ secretion was seen in the splenocytes of mice immunized with all formulations as compared to the control group (P<0.0001). In contrast, the lowest IL-4 production was detectable in Lip+imiquimod+SLA spleen, which was not significantly different compared with other groups. CONCLUSION: The results of this study show that liposome DOTAP+SLA+imiquimod formulation generates a cellular immune response that is protective against challenge against L. major.

The effect of Phase 1 periodontal treatment on the salivary RANKL/OPG ratio in severe chronic periodontitis.

BACKGROUND: RANKL and OPG play an important role in bone resorption. The R A N K L O P G ratio increases in periodontal disease. The present study was conducted to evaluate the effect of Phase 1 periodontal treatment on the salivary R A N K L O P G ratio in patients with severe chronic periodontitis. MATERIALS AND METHODS: Saliva samples were collected from 13 patients with severe chronic periodontitis and 14 healthy controls at baseline and then four weeks after the treatment using unstimulated spitting. The salivary levels of RANKL, OPG and R A N K L O P G ratio were investigated using the ELISA method. RESULTS: The findings of this study showed that the mean R A N K L O P G ratio was significantly higher in the patients with periodontal disease than in the healthy controls (P=0.001). Also, the R A N K L O P G ratio was significantly higher in the patients with a higher mean CAL (P=0/004). The comparison of the salivary R A N K L O P G ratio in patients with periodontal disease before and four weeks after the treatment showed a significant reduction in this value four weeks after the periodontal treatment (P=0/001). CONCLUSION: The results of this study showed a direct relationship between the R A N K L O P G ratio and the severity of periodontal disease. Phase 1 of periodontal treatment was found to be effective in reducing the R A N K L O P G ratio. In other words, the R A N K L O P G ratio can be a good predictor of treatment success. Further long-term studies with larger sample sizes are required for confirming these results.

Protection induced by Leishmania Major antigens and the imiquimod adjuvant encapsulated on liposomes in experimental cutaneous leishmaniasis.

There is a need for new, effective, and less expensive and toxic treatment for Leishmaniasis. It seems that the use of a suitable adjuvant and a delivery system is effective in inducing immune reactions for protection. Liposomes can be applied as immunoadjuvants to trigger immune reactions to different antigens. The adjuvant effects of imiquimod using DSPC liposomes containing SLA (soluble Leishmania antigens) were studied on the type and intensity of the produced immune reaction to the challenge of Leishmania major in BALB/c mice. Liposomes were produced by the lipid film procedure. BALB/C mice were immunized subcutaneously, three times at 2-week intervals and with various formulations. Lesion development and the parasite burden in the spleens and feet after the challenge with Leishmania major, Th1 cytokine (IFN-γ), and the IgG isotype titration were assessed to evaluate the induced immune reaction and the protection level. The group of mice immunized with Liposome DSPC +Imiquimod +SLA revealed less severe footpad swelling, being significantly different (P < .05) from other groups. A higher level of IgG2a and IFN-γ secretion was observed in the mice immunized with Liposome DSPC +Imiquimod +SLA than the control group. These observations imply that the DSPC liposome containing imiquimod induces the Th1 immune response that is protective against the challenge of Leishmania major.

Salivary MMP-1, MMP-2, MMP-3 and MMP-13 Levels in Patients with Oral Lichen Planus and Squamous Cell Carcinoma

The aim of present study was to evaluate salivary matrix metalloproteinase-1 (MMP-1), MMP-2, MMP-3 and MMP-13 levels in patients with oral lichen planus (OLP) and squamous cell carcinomas (SCC) as well as in healthy controls. Thirty cases of OLP (bilateral lesions, papular and reticular lesions, and Wickham lines) clinically and histopathologically (group A), 30 with oral SCCs (group B), and 30 with no history of oral cancer, other lesions or lichen planus (group C) were enrolled at the Department of Oral Medicine School of Dentistry, Zahedan, Iran. Unstimulated whole saliva was collected and laboratory measurement of salivary concentration of MMP-1, MMP-2, MMP-3 and MMP-13 was conducted by immuno-sorbent enzyme-linked methods. Data analysis was performed with Kruskal-Wallis and Mann-Whitney tests and Pearson's correlation coefficients. In the present study, MMP-2 and MMP-13 levels were higher in oral SCC patients than in OLP and healthy individuals. More research is required to assess MMP links with tumor invasion.

Imbalances in T Cell-Related Transcription Factors Among Patients with Hashimoto's Thyroiditis.

OBJECTIVES: Imbalances in effector T cell functioning have been associated with a number of autoimmune diseases, including Hashimoto's thyroiditis (HT). Differentiation of effector T helper (Th) 1, Th2, Th17 and regulatory T cell (Treg) lymphocytes is regulated by transcription factors, including Th1-specific T box (T-bet), GATA binding protein-3 (GATA3), retinoid-related orphan receptor (ROR)-α and forkhead box P3 (FOXP3). This study aimed to investigate Th1/Th2, Th1/Treg, Th2/Treg and Th17/Treg balances at the level of these transcription factors. METHODS: This study took place between October 2015 and August 2016. Peripheral blood mononuclear cells were collected from a control group of 40 healthy women recruited from the Zahedan University of Medical Sciences, Zahedan, Iran, and a patient group of 40 women with HT referred to the Hazrat Ali Asghar Hospital, Zahedan. Total ribonucleic acid extraction was performed and the gene expression of transcription factors was quantitated using a real-time polymerase chain reaction technique. RESULTS: Expression of T-bet and GATA3 was significantly elevated, while FOXP3 expression was significantly diminished among HT patients in comparison with the controls (P = 0.03, 0.01 and 0.05, respectively). Expression of RORα was higher among HT patients, although this difference was not significant (P = 0.15). Expression of T-bet/FOXP3, GATA3/FOXP3 and RORα/FOXP3 ratios were increased among HT patients in comparison with the controls (P <0.02, <0.01 and <0.01, respectively). CONCLUSION: These results indicate that HT patients have imbalances in Th1/Treg, Th2/Treg and Th17/Treg lymphocytes at the level of the transcription factors, deviating towards Th1, Th2 and Th17 cells. Correction of these imbalances may therefore be therapeutic.

Association between interleukin-1 receptor associated kinase 1 rs3027898 A/C gene polymorphism and rheumatoid arthritis.

Rheumatoid arthritis (RA) is a systemic autoimmune disease predominantly involving the synovial joints and affects up to 1% of adults worldwide. The aim of the present study was to determine whether the interleukin-1 receptor (IL1R)-associated kinase (IRAK1) rs3027898 gene polymorphism confers susceptibility to RA in a sample of patients from Iran. This gene encodes IRAK1, one of two putative serine/threonine kinases that associates with IL1R upon stimulation. IRAK1 is partially responsible for IL-1-induced upregulation of the transcription factor, nuclear factor-κB. The present case-control study was performed on 120 patients with RA and 120 healthy individuals. Genomic DNA was extracted from whole blood, and the gene polymorphism was evaluated using a tetra-primer amplification refractory mutation system-polymerase chain reaction method. The results demonstrated that there was no association between IRAK1 rs3027898 CA genotype and the risk of RA in women (odds ratio=0.72, 95% confidence interval=0.41-1.49; P=0.446). Further studies with larger sample sizes and different ethnicities are required to validate the present findings.

Lectin, galactoside-binding, soluble, 3 rs4652 A/C gene variation and the risk for rheumatoid arthritis.

Rheumatoid arthritis (RA) is a complex genetic disease. The lectin, galactoside-binding, soluble, 3 (LGALS3) gene, encodes a member of the galectin family of carbohydrate binding proteins, and is one of the best examples of a non-human leukocyte antigen gene associated with a risk for RA in various populations. In the current study, the association between LGALS3 rs4652 gene polymorphism and RA was examined. This case-control study was performed on the 120 patients with RA and 120 healthy subjects. Genomic DNA was extracted from whole blood, and gene polymorphism was tested using a tetra-primer amplification refractory mutation system-polymerase chain reaction. The results demonstrated that LGALS3 rs4652 AC genotype increased the risk of RA (OR=11.622, 95% CI=4.473-28.656; P=0.001) when compared with the AA genotype. However, the CC genotype and the C allele were not associated with RA. These findings indicated an association between LGALS3 rs4652 variation and the risk of RA in a sample of Iranian individuals. Further studies with larger sample sizes and populations of different ethnicities are required to validate our findings.

Investigating the level of salivary endothelin-1 in premalignant and malignant lesions.

PURPOSE: This study aimed to investigate the level of salivary endothelin-1 in premalignant and malignant lesions. MATERIALS AND METHODS: In this study, 75 cases were investigated of which 25 cases were healthy, 25 cases had oral lichen planus (OLP), and 25 cases had oral squamous cell carcinoma (OSCC). In order to collect the saliva samples, the unstimulated spitting was used. The samples were collected between 9 and 12 a.m. They were sent to the lab shortly after being collected and salivary endothelin-1 was recorded for each sample according to the instruction of factory by using enzyme-linked immunosorbent assay and optical density at a wavelength of 450 nm. SPSS version 20 and one-way ANOVA and LSD tests were used to analyze the data. RESULTS: The mean of salivary endothelin-1 level in patients with OSCC was 163.98 pg/ml, in patients with OLP was 160.9 pg/ml, and in healthy people was 137.19 pg/ml. The analysis of one-way ANOVA suggested that the level of salivary endothelin-1 in both groups was the same and significantly higher than that in control group (p < 0.05). CONCLUSIONS: The level of salivary endothelin-1 in patients with SCC and OLP was higher than that in healthy group. Thus, it can be used as the latest therapeutic protocol for oral premalignant and malignant lesions.

Comparison of Cotinine Salivary Levels in Hookah Smokers, Passive Smokers, and Non-Smokers.

BACKGROUND: At present smoking is considered a great health-related problem. Smoking cigarettes and use of tobacco are on the rise in the Middle East countries; therefore, the number of people exposed to passive cigarette smoke is increasing, too. The aim of the present study was to determine and compare salivary cotinine levels in hookah smokers, individuals exposed to passive cigarette smoke and non-smoker (passive smokers). METHODS: In the present cross-sectional study, unstimulated salivary samples were collected from 150 subjects, including 50 hookah smokers, 50 passive smokers, and 50 non-smokers. Bioassay Technology Laboratory cotinine kit was used to determine salivary levels of cotinine using the enzyme-linked immunosorbent assay (ELISA) technique at a sensitivity rate of 0.019 pg/ml. Data were analyzed with SPSS software using t-test and Pearson's correlation coefficient. FINDINGS: The highest salivary cotinine levels were recorded in hookah users (20.24 ± 5.62 ng/ml), followed by passive smokers (16.09 ± 3.51 ng/ml), in descending order. No detectable cotinine levels were observed in non-smokers. Pearson's correlation coefficient showed a strong and positive correlation between use of hookah and salivary cotinine levels (r = 0.932, P = 0.001). CONCLUSION: Based on the results of the present study, salivary cotinine levels were higher in hookah smokers compared with passive smokers and non-smokers, in descending order.

Patho-TB test for the rapid diagnosis of pulmonary tuberculosis.

BACKGROUND: Despite recent technologic improvements in identifying mycobacterium tuberculosis, we are still facing problems in rapid diagnosis of tuberculosis. The objective of this study is to determine the diagnostic value of a new rapid screening test (Patho-TB™) for diagnosis of pulmonary tuberculosis. METHODS: Between September 2006 to August 2007, 178 patients were enrolled in the study who were finally classified into two groups; a group of documented pulmonary tuberculosis (n = 67) and a group of non-tuberculous pulmonary infection (n = 111). Patho-TB™ test, Ziehl-Neelsen staining and culture were done on all specimens. RESULTS: Of all, 43 patients with pulmonary tuberculosis were sputum smear positive for acid fast bacilli and the rest were smear negative. Mean age of the patients was 59.8 ± 16.1 years and 44% of them were men. The results of Patho-TB™ test were positive in 40 of smear positive and 20 of smear negative tuberculous patients and 33 cases of non-tuberculous control group. The sensitivity, specificity, positive and negative predictive values and accuracy of Patho-TB™ test were estimated 89.5%, 70.2%, 64.5%, 91.7% and 77.5%, respectively. CONCLUSIONS: According to the present study it would be suggested that Patho-TB™ test could be a rapid and inexpensive method for diagnosis of pulmonary tuberculosis, given by its high sensitivity and negative predictive value. Concerning the high number of false positive results, using a confirmatory diagnostic procedure is mandatory.