Molecular Genotyping of Giardia duodenalis in Humans in the Yazd County, Central of Iran.

Background: We aimed to investigate the molecular genotyping of Giardia duodenalis in Humans in the Yazd County, Central, Iran. Methods: Total of 35 fecal samples were collected from patients referred to Yazd Central Laboratory, Yazd, Iran from February to July 2022. All the samples were included in this study after microscopic observation of G. duodenalis. DNA samples were extracted using related kit and were analyzed by Nano Drop. The molecular assessment was carried out using semi-nested PCR using the target gene of gdh. All amplified samples were sequenced using Sanger method. BLAST analyzed the sequences for assemblage identification. Results: Out of 35 samples, 24 (68.57%) and 11 (31.43%) were male and female, respectively. All included samples were amplified using the specific gdh primer pair. The molecular analysis showed 17 isolates (48.57%) as assemblage BIV, 8 isolates (22.86%) as assemblage BIII, 6 isolates (17.14%) as assemblage AII and 4 isolates (11.43%) as assemblage AIII (P<0.05). Conclusion: Assemblages A and B are the most prevalent in Central Iran. The molecular identification of G. duodenalis isolates from animals and implementing control programs.

Expression profile in Leishmania major exposed to Staphylococcus aureus and group A beta-hemolytic Streptococcus.

BACKGROUND & OBJECTIVES: The interaction of Leishmania spp. with microbiota inside the midgut vector has significant output in pathogenesis. This study aimed to identify the profile of Leishmania majorgene expression of LACK, gp63, and hsp70after exposure to Staphylococcus aureusand group A beta-hemolytic Streptococci (GABHS). METHODS: Leishmania major (MRHO/IR/75/ER) promastigotes were exposed with S. aureus, with GABHS, and with both GABHS and S. aureus at 25°C for 72 h. The gene expression analysis of Lmgp63, Lmhsp70,and LmLACKwas assessed using SYBR Green real-time PCR by ΔΔCt. All experiments were repeated in triplicate. Statistical analysis was done using two-way ANOVA. A P-value less than 0.05 was considered significant. RESULTS: Lmgp63 was expressed in the group exposed to GABHS with 1.75-fold lower than the control group (p=0.000). The LmLACK had expression in both groups exposed with GABHS and GABHS with S. aureus with 2.8 and 1.33-fold more than the control group, respectively (p=0.000). The Lmhsp70 gene expression was reported in the group exposed with GABHS with relative quantification of 5.7-fold more than the control group. INTERPRETATION & CONCLUSION: This study showed that the important genes encoding LACK, gp63, and hsp70 changed their expression after exposure to the S. aureus and GABHS.

Safety assessment of genetically modified rice expressing Cry1Ab protein in Sprague-Dawley rats.

Rice is considered one of the most important staple food crops. Genetically modified (GM) Bt rice, harbored cry1Ab gene expressing the insect-resistance protein has been developed to resistance to the insects. In this study, we assessed the safety of the GM Bt rice on Sprague-Dawley rats for 90 days. Totally, 120 rats in both sexes were used for three different diets, including 50% GM Bt rice, feeding with 50% rice, and standard feeding. Each 40 SD rats including 20 males and 20 females were considered as each diet. The clinical variables such as body weight and food consumption were measured and a range of clinical tests was examined, including hematology, serum chemistry parameters, urinalysis profile, thyroid, and sex hormone levels. Pathological assessments were also done. The results showed that the mean weekly feed utilization (%) had no significant difference among the studied groups. Also, blood biochemistry, hematological parameters, urine analysis, and hormonal levels had no significant differences among the groups. However, alanine aminotransferase was less in males versus female feeding with GM Bt rice. No histopathological changes were observed among the groups. In conclusion, this study demonstrated that GM Bt rice had no obvious adverse effects on rats' health.

Isolation and Molecular Identification of Leishmania spp. Agents in Patients with Cutaneous Leishmaniasis in Yazd Province, Endemic Region of Central Iran.

BACKGROUND: Cutaneous Leishmaniasis (CL) is a major health problem in many parts of Iran. Many methods have been introduced for detection and identification of Cutaneous Leishmaniasis. The purpose of this study was isolation and molecular identification of Leishmania spp. agents in patients with CL from endemic region of central Iran. In this study, one of the main loci of central Iran named Yazd will be assessed CL identification using PCR-RFLP. METHODS: For this cross-sectional study, sampling was done from 372 suspicious patients with CL who referred to Health Centers of Yazd Province from 2016 to 2017. After collection samples of patients, DNA extraction was done from samples on slides. Genus detection was done using specific primers by PCR. RFLP analysis was done for species identification. RESULTS: Out of 372 samples, 159 samples were positive using PCR based method. Out of 159 samples, 87 (54.7%) L. major and 72 (45.3%) L. tropica were identified using RFLP analysis. The number of lesions in each patient was different but 119 (74.8%) patients showed the number of 1-3 lesions, and more lesions (more than 10 lesions) was showed in 4 (2.5%) person. CONCLUSION: The CL found in Yazd province resulted from L. major and L. tropica as the agents of rural and urban types, respectively. The prevalence of L. major and L. tropica was almost the same. This indicated that control programs could be designed for treatment and vector and reservoir control.

Applying Rogers' framework to evaluate public awareness and knowledge of medical genetics in a developing country.

Public knowledge of medical genetics is essential for better establishment of its services but has been rarely evaluated based on distinguished types of knowledge. We designed and validated a new self-administered questionnaire in Farsi (Persian language) to assess public knowledge of medical genetics based on Rogers' framework. This framework divides knowledge into three types of awareness, how-to (practical) and principles knowledge which refer to knowing the existence, proper use, and theoretical principles of an innovation, respectively. We asked consecutive individuals (n = 306, age ≥ 20 years) visiting health centers in different regions of Yazd, a city in central Iran, to fill out the questionnaire. After validation, we analyzed 280 of the questionnaires which revealed a high degree of internal consistency (Cronbach's alpha 0.90) and a positive linear relationship among the scores of different knowledge. Our respondents had relatively fair awareness and how-to, but generally poor principles knowledge with statistically significantly better scores in females and those with higher education. We observed tangible strengths in topics such as consanguineous marriage, thalassemia, and hereditary predisposition to diabetes and cardiovascular disorders, and weaknesses in areas such as genetic testing and genetics of cancer. Notably, experience of premarital genetic counseling did not show any significant effect, but having a relative with a genetic disorder was significantly linked to better awareness scores. Our study provides a reliable and self-administered questionnaire for the assessment of public knowledge of medical genetics. Despite revealing important strengths and weaknesses in our population sample, larger scale evaluations in Iran and other developing countries are needed for better understanding of the public knowledge as the prerequisite for designing appropriate educational programs.

J-binding protein 1 and J-binding protein 2 expression in clinical Leishmania major no response-antimonial isolates.

Cutaneous leishmaniasis (CL) is a major disease in many parts of the world. Since no vaccine has been developed, treatment is the best way to control it. In most areas, antimonial resistance whose mechanisms have not been completely understood has been reported. The main aim of this study is gene expression assessing of J-binging protein 1 and J-binding protein 2 in clinical Leishmania major isolates. The patients with CL from central and north Iran were considered for this study. The samples were transferred in RNAlater solution and stored in - 20 °C. RNA extraction and cDNA synthesis were performed. The gene expression analysis was done with SYBR Green real-time PCR using ∆∆CT. Written informed consent forms were filled out by patients, and then, information forms were filled out based on the Helsinki Declaration. Statistical analysis was done with SPSS (16.0; SPSS Inc, Chicago) using independent t test, Shapiro-Wilk, and Pearson's and Spearman's rank correlation coefficients. P ≤ 0.05 was considered significant. The gene expression of JBP1 and JBP2 had no relation with sex and age. The JBP1 gene expression was high in sensitive isolates obtained from north of the country. The JBP2 gene expression was significant in sensitive and no response-antimonial isolates from the north, but no significant differences were detected in sensitive and resistant isolates from central Iran. Differential gene expression of JBP1 and JBP2 in various clinical resistances isolates in different geographical areas shows multifactorial ways of developing resistance in different isolates.

The Effect of Hyoscine on Pain, Tenderness, and Rebound Tenderness in Patients with Appendicitis: Quasi-Interventional Study.

OBJECTIVE: To evaluate the effect of hyoscine on pain and tenderness, rebound tenderness and appendicitis patients before surgery and this study can be considered as the first study investigated the efficacy of hyoscine on pain, tenderness, and rebound tenderness in patients with appendicitis. METHODS: In this single-group, prospective interventional study (before-after) according to inclusion and exclusion criteria, 70 patients with pain in the right lower abdomen and typical symptoms of appendicitis were evaluated and after surgery, diagnostic accuracy was examined by pathologic results. The pain was evaluated before and after administration of hyoscine by numerical and verbal examination. Tenderness and rebound tenderness were also determined. RESULTS: The mean age of patients was 26.81±7.66. Totally, 42 patients (60%) had reduction in pain, 50 patients (71% percent) had reduction in tenderness, and 39 (55%) had reduction in rebound tenderness after treatment with the drug. A statistically significant reduction of pain and tenderness, rebound tenderness was observed in all of men after administration of hyoscine (p<0.001) but in women, the reduction of severity of tender results was only significant (p=0.002). Data analysis in women and men together showed the significant reduction of pain, tenderness, and rebound tenderness (p<0.001). CONCLUSION: According to the results, hyoscine can reduce pain, tenderness, and rebound tenderness. So, it seems that hyoscine is a good candidate for patients with appendicitis. IRCT REGISTRATION NUMBER: IRCT2015111825123N1.

Polyurethane sheet impregnated with Arabinogalactan can lead to increase of attachment of promastigotes and Amastigote of Leishmania major (MRHO/IR/75/ER) by GP63 and HSP70 genes.

The aim of this study was to investigate the effect of polyurethane sheet (PUS) and polyurethane sheet impregnated with Arabinogalactan (PUSIAG) on the cell attachment and viability of Promastigotes and Amastigotes of Leishmania major (MRHO/IR/75/ER), and mouse macrophages, and its whole skin cells (WSCs). In a sterile condition, 10 mL of Arabinogalactan 5% w/v was poured into a falcon. Then, a piece of PUS was placed inside it, and incubated at 37 °C for 24 h. Next, it was washed, and cut. Then, one piece of PUS and PUSIAG was separately added to 1 mL of cell suspension (Promastigotes, Amastigote, and WSCs), and then incubated for 1, 2, 3, and 4 days at 37 °C. After incubation times, the quantity of adhered cells was counted, and cell viability was measured by MTT assay. Also, for WSCs and macrophages, the expression of integrin, fibronectin and GAPDH was investigated, and for Promastigotes and Amastigotes, the expression of GP63, Cpb, and 18s rRNA was measured. This study showed that with increase of exposure time, the percentage of attached cells was increased. There was a significant difference between attached cells to PUSIAG and PUS in case of Promastigotes and Amastigotes. It seems that Promastigotes and Amastigotes have higher interest to PUSIAG than WSCs and Macrophages. Also, this study showed with increase of exposure time, the percentage of viable cells was decreased. There were significant differences between cell viability of Promastigotes and Amastigotes when exposed to PUSIAG and PUS, especially in long time incubation. Also, when incubation time was increased the relative expression of integrin and fibronectin in WSCs and macrophages, and GP63 and HSP70 in Promastigotes and Amastigotes were increased.

Aquaglyceroporin1 gene expression in antimony resistance and susceptible Leishmania major isolates.

BACKGROUND & OBJECTIVES: The mechanism of antimony resistance in Leishmania has been studied extensively, in connection with decreased influx and/or increased eflux of the drug. Aquaporin 1 (AQP1) protein has been shown to mediate the uptake of trivalent antimony. This study was aimed to find the expression level of AQP1 gene in resistant versus non-resistant clinical isolates of Leishmania major in Iranian patients. METHODS: Clinical isolates were obtained from 16 considered patients referred to Navab Safavi Clinical Center, Isfahan, Iran from October 2014 to December 2015. After diagnosis of cutaneous leishmaniasis using microscopic observation, biopsy was performed from lesion(s) of each patient and stored inside RNAlater solution at -20΀C. Written informed consent was obtained from all the patients to participate in the study before recording their information and sampling based on Helsinki declaration. Each patient was treated with Glucantime and followed for three months. All sensitive and resistance isolates were considered and compared with AQP1 gene expression using real time PCR that was analyzed with delta-delta Ct. RESULTS: Out of 16 clinical isolates, four patients were resistant and 12 were non-resistant. The AQP1 gene expression in resistant isolates was significantly higher than the one in response failure isolates (p = 0.001). INTERPRETATION & CONCLUSION: The significant over expression (0.5 fold) of AQP1 gene in resistant versus non- resistant isolates suggests different mechanism of drug resistance such as mutations. Mutations may change the physiological function of the Aquaporin 1 protein that might affect its expression level.

Ultrasound Assistance In Differentiating Malignant Thyroid Nodules From Benign Ones.

BACKGROUND: The search is ongoing for simple, effective ways to identify and differentiate thyroid nodules in order to avoid invasive procedures. This study aims to perform an ultrasound assessment of clinically suspected patients with malignant nodules, to perform a fine needle biopsy on them, and to compare the results obtained. METHODS: In total, 135 patients with thyroid nodules suspected to be malignant in the ultrasound underwent ultrasound-guided fine needle biopsies. The patients' gender, age, ultrasound views (echogenicity, shape, and calcification type), nodule size, number of nodules, and needle biopsy results were retrospectively evaluated. RESULTS: Of the 135 patients, 117 (86.7%) were female and 18 (13.3%) were male. In terms of age, 67 (49.16%) were younger than 40 and the rest were older. According to the Chi-square test, a significant relationship was found between the type of nodule and calcification found in the ultrasound views (p=0.001). The nodule type was not significantly related to gender (p=0.563) or to the number of nodules (p=0.128); however, there was a significant relationship between nodule size and type (p=0.001). Ultrasound specificity, sensitivity, positive and negative predictive values, and accuracy for differentiating benign from malignant nodules were 93.2%, 93.8%, 81.1%, 98%, and 93.3%, respectively. CONCLUSIONS: Ultrasound views (comet tail artefact and linear echogenic foci) were better predictors of benign nodules than of malignant ones, while round, echogenic foci, brighter and larger than typical micro-calcifications without any visible echoes and multiple punctuate round echogenic foci were better predicted malignancy.

CLINICOPATHOLOGICAL FEATURES OF LIVER TUMOURS: A TEN-YEAR STUDY.

BACKGROUND: Various diseases affect the liver, among them, malignant and benign tumours with hepatic nodules are the most important. We aimed to evaluate the clinicopathological findings related to hepatic tumours and nodules. METHODS: This retrospective study was carried out during November 2014 to August 2015 by reviewing the hospital medical records of 164 registered patients with liver biopsies referred to Shahid Sadoughi educational General Hospital, Yazd, Iran, between 2004 and 2014. The samples were selected through the census method. Age, gender, clinical symptoms, initial clinical diagnosis, pathology reports and ultrasound results were considered as variables. Data were analysed by using SPSS-17. RESULTS: There were 87 (53%) men and 77 (47%) women. The mean ages of presentation for malignant and benign tumours were 57.9 ± 17.2 and 44.9 ± 19.4 years, respectively. Seventy benign tumours and 147 malignant tumours were recorded. The most frequent chief complaint was abdominal pain (54.9%) in both malignant (56.50%) and benign tumours (41.20%). Hepatocellular carcinoma (HCC) and hemangioma were the most prevalent malignant and benign hepatic tumours, respectively. In our study, correlation between pathology reports and primary diagnoses was 40.9%, and a significant relationship was found between sonography and pathological findings (p = 0.038). CONCLUSIONS: We found that only when primary clinical diagnosis and sonography were in favour of malignancy, they were correlated with pathology results. Clinicopathological assessments can help physicians in their diagnosis in order to facilitate the management of hepatic tumours.