Cloning, expression and purification of pwo polymerase from pyrococcus woesei

Background and objectives: Pyrococcus woesei is a hyperthermophilic archaea and produces a heat stable polymerase (Pwo polymerase) that has proofreading activity. Materials and Methods: In this study, this microorganism was cultured, its DNA was extracted and the pwo gene polymerase was cloned, expressed and purified. The DNA sequence of the cloned gene was verified by sequencing. The pwo polymerase gene consists of 2,328 bps (775 amino acids with about 90 kD molecular weight). Cloning was done by GATEWAY TM Cloning System and for purification of recombinant protein; His6x-Tag was added to the C-terminus of the recombinant protein. Results and Conclusion: We could purify Pwo polymerase enzyme by Ni-NTA resin. PCR assay showed that Pwo polymerase activity is comparable to a commercial Pfu polymerase activity

Acne in traditional persian medicine

Background: Acne Vulgaris is a common skin condition in 35-90 of youth. Although traditional methods of treating acne in Iran seem to be common and high demand, a few studies about this issue have been accomplished. This study aimed to find out how much definitions and recommendations about this condition in Traditional Persian Medicine (TPM) are similar to contemporary medicine. Methods: Seven main reference books from TPM were selected. The equivalents of acne in TPM were found and comparison of the definitions and recommendations of TPM and CM were extracted. Two diseases called Namlah and Bosoor-Labanieh were most similar to acne. These keywords were searched in TPM Books by using �Noor software�. Results: In TPM references, two diseases of �Namlah� and �Bosoor-Labanieh� are very similar to acne. The cause of these diseases in TPM textbooks has been explained as the effort of the body for the disposal of wastes through the skin and accumulation of them below it. The treatments are lifestyle modification, drug therapy, and manual interventions. Conclusion: Interventions of TPM have been experienced by Iranian physicians of the Old Testament over hundreds of years. There are meaningful similarities among TPM texts in descriptions and advice. In addition, there are remarkable similarities in the disease characteristics and some of the recommendations for acne in TPM and CM. The effectiveness and cost-benefit of the recommended TPM therapeutics and methods can be considered and evaluated as hypotheses for the case and group studies and clinical trials. © 2020, Iranian Journal of Public Health. All rights reserved

The effects of myrtle (myrtus communis) and clindamycin topical solution in the treatment of mild to moderate acne vulgaris: A comparative split-face study

Objectives: Although Acne vulgaris is a chronic skin disease, which its standard treatment causes therapeutic limitations and some common adverse effects, medicinal plants can be effective in treatment with low adverse effects as combination therapy. Myrtle (Myrtus Communis) has some beneficial properties, which has been administered topically and orally for some skin diseases in Persian medicine. This study aimed to compare the efficacy and safety of Myrtle formula and 1 clindamycin topical solution. Methods: This was a split-face clinical trial that was done on 55 patients with mild to moderate acne vulgaris for 16 weeks. The patients received topical Myrtle solution to the right side of the face (group 1) and clindamycin solution to the left side (group 2) twice daily for 12 weeks. All participants were examined for the acne severity index (ASI) and total acne lesions counting (TLC) at certain times during the study. Then, they stopped using them for four weeks. They also did not take the drug in the final four weeks of the study. Results: Forty-eight patients completed the study for 16 weeks; 40 (83.2) patients were female and the rest of them were male. The mean age and standard deviation were 25.62 ± 7.62 years. After 12 weeks, the percentage changes of comedones, inflammatory lesions, ASI and TLC were significantly reduced in both groups (p < 0.001). The percentage change of inflammatory lesions and ASI decrease was significantly higher in the group 1 (p = 0.03). There was no significant difference in the incidence of side effects between the two groups. There was a more significant decrease in sebum percentage change in the group 1 (p = 0.003). Conclusion: Myrtle lotion was effective and safe for the treatment of mild to moderate acne vulgaris. Copyright © Korean Pharmacopuncture Institut

Psychiatric disorders in Iranian children and adolescents

Objective: The aim of the present study was to investigate the epidemiology of psychiatric disorders in children and adolescents in five provinces of Iran: Tehran, Shiraz, Isfahan, Tabriz and Mashhad. Method: In the present study, we selected 9,636 children and adolescents aged 6-18 years through multistage cluster random sampling method from Tehran, Shiraz, Isfahan, Tabriz and Mashhad. We instructed the clinical psychologists to complete the Strengths and Difficulties Questionnaire (SDQ) for the participants, and those who received a high score on SDQ, completed the Persian version of Kiddie-SADS-Present and Lifetime Version (K-SADS-PL). We used descriptive analysis and 95 confidence interval to investigate the relationship between scores of the K- SADS questionnaire and demographic factors . We used one-way ANOVA to test the significant differences among the disorders according to sex, age and province of residence. Results: Based on the results, oppositional defiant disorder (ODD) (4.45) had the highest prevalence of psychiatric disorders in the five provinces and substance abuse and alcohol abuse (0) had the lowest prevalence. In addition, attention deficit hyperactivity disorder (ADHD) had the most prevalence in boys (5.03) and ODD had the most prevalence in girls (4.05). Among the three age groups, 6 to 9 year olds had the highest rates of ADHD (5.69); 10 to 14 and 15 to 18 year olds had the highest rates of ODD (4.32 and 4.37 respectively). Among the five provinces, Tehran and Mashhad allocated the highest rates of ODD; Isfahan and Shiraz had the highest rates of ADHD; and Tabriz had the highest rates of social phobia. Conclusion: The current study revealed that the overall frequency of psychiatric disorders based on Kiddie- SADS-Present and Lifetime Version (K-SADS-PL) was higher than a similar study. Moreover, in this study, among the five provinces, Tehran and Mashhad allocated the highest rates of ODD; Isfahan and Shiraz had the highest rates of ADHD; and Tabriz had the highest rates of social phobia. Therefore, these percentage of psychiatric disorders in Iran lead us toward a greater use of consultation and mental health services

Psychological problems of Iranian children and adolescents: Parent report form of Strengths and Difficulties Questionnaire

Objective: The aim of this study was to evaluate the degree of psychological problems Iranian children and adolescents have, using parent report form of the Strengths and Difficulties Questionnaire (SDQ). Methods: In a community-based study, 9636 children and adolescents aged 6-17 years were selected using the multistage cluster random sampling method from five provinces of Iran: Tehran, Isfahan, Fars, Razavi Khorasan and East Azerbaijan. The parents completed the SDQ, which consisted of five subscales including emotional problems, conduct problems, hyperactivity, peer problems and prosocial behaviors. Result: The results revealed 21.4 of emotional problems, 32.9 of conduct problems, 20 of hyperactivity, 25.6 of peer problems, 7.6 of problems in prosocial behaviors and 16.7 of total difficulties among Iranian children and adolescents. We found that emotional problems were more prevalent among girls, while conduct problems, hyperactivity, total difficulties and problems in prosocial behaviors were more prevalent among boys. High educational level of parents was a protective factor against some psychological problems. Conclusion: Considering the proportion of psychological problems in Iranian children and adolescents, we need to develop and implement special policies and programs to provide appropriate mental health services. © 2014 Shadowfax Publishing and Informa UK Limited

Psychiatric disorders in Iranian children and adolescents

Objective: The aim of the present study was to investigate the epidemiology of psychiatric disorders in children and adolescents in five provinces of Iran: Tehran, Shiraz, Isfahan, Tabriz and Mashhad. Method: In the present study, we selected 9,636 children and adolescents aged 6-18 years through multistage cluster random sampling method from Tehran, Shiraz, Isfahan, Tabriz and Mashhad. We instructed the clinical psychologists to complete the Strengths and Difficulties Questionnaire (SDQ) for the participants, and those who received a high score on SDQ, completed the Persian version of Kiddie-SADS-Present and Lifetime Version (K-SADS-PL). We used descriptive analysis and 95 confidence interval to investigate the relationship between scores of the K- SADS questionnaire and demographic factors . We used one-way ANOVA to test the significant differences among the disorders according to sex, age and province of residence. Results: Based on the results, oppositional defiant disorder (ODD) (4.45) had the highest prevalence of psychiatric disorders in the five provinces and substance abuse and alcohol abuse (0) had the lowest prevalence. In addition, attention deficit hyperactivity disorder (ADHD) had the most prevalence in boys (5.03) and ODD had the most prevalence in girls (4.05). Among the three age groups, 6 to 9 year olds had the highest rates of ADHD (5.69); 10 to 14 and 15 to 18 year olds had the highest rates of ODD (4.32 and 4.37 respectively). Among the five provinces, Tehran and Mashhad allocated the highest rates of ODD; Isfahan and Shiraz had the highest rates of ADHD; and Tabriz had the highest rates of social phobia. Conclusion: The current study revealed that the overall frequency of psychiatric disorders based on Kiddie- SADS-Present and Lifetime Version (K-SADS-PL) was higher than a similar study. Moreover, in this study, among the five provinces, Tehran and Mashhad allocated the highest rates of ODD; Isfahan and Shiraz had the highest rates of ADHD; and Tabriz had the highest rates of social phobia. Therefore, these percentage of psychiatric disorders in Iran lead us toward a greater use of consultation and mental health services

Mice Orally Immunized with a Transgenic Plant Expressing the Glycoprotein of Crimean-Congo Hemorrhagic Fever Virus ▿

While Crimean-Congo hemorrhagic fever (CCHF) has a high mortality rate in humans, the associated virus (CCHFV) does not induce clinical symptoms in animals, but animals play an important role in disease transmission to humans. Our aim in this study was to examine the immunogenicity of the CCHFV glycoprotein when expressed in the root and leaf of transgenic plants via hairy roots and stable transformation of tobacco plants, respectively. After confirmatory analyses of transgenic plant lines and quantification of the expressed glycoprotein, mice were either fed with the transgenic leaves or roots, fed the transgenic plant material and injected subcutaneously with the plant-made CCHFV glycoprotein (fed/boosted), vaccinated with an attenuated CCHF vaccine (positive control), or received no treatment (negative control). All immunized groups had a consistent rise in anti-glycoprotein IgG and IgA antibodies in their serum and feces, respectively. The mice in the fed/boosted group showed a significant rise in specific IgG antibodies after a single boost. Our results imply that oral immunization of animals with edible materials from transgenic plants is feasible, and further assessments are under way. In addition, while the study of CCHF is challenging, our protocol should be further used to study CCHFV infection in the knockout mouse model and virus neutralization assays in biosafety level 4 laboratories

A protective cocktail vaccine against murine cutaneous leishmaniasis with DNA encoding cysteine proteinases of Leishmania major.

The protection elicited by the intramuscular injection of two plasmid DNAs encoding Leishmania major cysteine proteinase type I (CPb) and type II (CPa) was evaluated in a murine model of experimental cutaneous leishmaniasis. BALB/c mice were immunized either separately or with a cocktail of the two plasmids expressing CPa or CPb. It was only when the cpa and cpb genes were co-injected that long lasting protection against parasite challenge was achieved. Similar protection was also observed when animals were first immunized with cpa/cpb DNA followed by recombinant CPa/CPb boost. Analysis of the immune response showed that protected animals developed a specific Th1 immune response, which was associated with an increase of IFN-gamma production. This is the first report demonstrating that co-injection of two genes expressing different antigens induces a long lasting protective response, whereas the separate injection of cysteine proteases genes is not protective

Humoral and cellular immune responses against Type I cysteine proteinase of Leishmania infantum are higher in asymptomatic than symptomatic dogs selected from a naturally infected population.

Canids are natural reservoirs of Leishmania infantum and have been promoted as experimental hosts to decipher the pathogenesis of human visceral leishmaniasis (VL). In this study, the presence of IgG antibodies as well as the presence of mononuclear leukocytes reactive to different cysteine proteinases (CPs) were examined in 13 L. infantum-infected dogs (six with symptoms, seven asymptomatic). Cysteine proteinases which belong to papain-like enzymes known as clan CA are the most studied CPs of parasite protozoa. These molecules are expressed by the intracellular stages of the parasite and could be immunogenic. We studied Type II CP (CPA) and Type I CP (CPB) with its long C-terminal extension (CTE) which could be highly immunogenic. We showed that the level of antibodies reactive to rCPA is low in both symptomatic and asymptomatic dogs. In contrast, when CPB and CTE were used as antigens, the level of total IgG (with IgG2 superior to IgG1) reached higher values in asymptomatic dogs than in dogs with VL. While the peripheral blood mononuclear cell (PBMC) reactivity was significant when cultured in the presence of freezed/thawed (F/T) lysate, it remained low in presence of CP although always higher for PBMC recovered from asymptomatic dogs. We showed the importance of CPB and CTE in particular as a target of immune response and their potential use for serodiagnosis in asymptomatic dogs