فهرست مطالب محسن واعظ

One of the most important hosts used for production of recombinant proteins is Escherichia Coli. For example, the most therapeutic proteins approved by FDA are produced in Escherichia Coli. Comprehensive knowledge about biologic nature of Escherichia Coli had made this microorganism to a favorable factory for production of recombinant proteins. Accessibility of this information have led to rational manipulation for changing of this small factory to intelligent system can make different recombinant proteins easier. So that, many engineered and useful strains were obtained from wild type and parental strains can produce high amount of diverse and stable recombinant proteins in lab and industrial scale. In this review, we will present some of these strains that are more widely used.

 The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is still an ongoing challenge in health system worldwide. Molecular detection methods including reverse transcription loop-mediated isothermal amplification (RT-LAMP) can help cutting off the transmission chain of the virus. In this study, prompt setting up of the RT-LAMP assays were investigated using gene construct design approach.

 RT-LAMP assays were initially setup and optimized using gene constructs as positive controls which worked simultaneously for RT-LAMP and RT-PCR assays. Constructed genes included T7 promoter sequence and partial sequences of SARS-CoV-2 RdRp, N and E genes cloned into pUC57 multiple cloning site (MCS) via synthetic and subcloning procedures. These templates were then used for artificial RNA synthesis. Finally, the RT-LAMP assay parameters were optimized and applied to clinical RNA specimens.

As an initial rapid approach, RT-LAMP assays were successfully setup for SARS-CoV-2 diagnostic using in vitro RNA transcripts from gene constructs. The assays were then examined on clinical samples with reaction times of 35 and 40 min required for assay one and two respectively. The detection limit was 100 copies of the target gene per reaction for each assay.

 Here, prompt setting up of RT-LAMP assays was carried out initially on designed gene constructs containing different SARS-CoV-2 target genes and simultaneous validation by RT-PCR. This approach could be used as an efficient solution where the clinical specimens may not initially be available, with the feasibility for new target gene of interest to be inserted into the MCS position.

The purpose of this research is to evaluate the leisure time of students Razavi University of Islamic sciences with emphasis on sport activities.

The present study was applied in terms of purpose and in the category of descriptive survey research that conducted in the field. The statistical community included male students of Razavi University of Islamic Sciences, 290 of whom were selected by random sampling. A questionnaire was used to collect data. The validity of the questionnaire was confirmed by experts and its reliability with using Cronbach's alpha was obtained above 0.7. Descriptive and inferential statistics such as Friedman test, one-sample t-test in SPSS software version 26 were used.

The test results showed that two factors, mental-emotional and hygienic health respectively, have a positive and significant effect on students' satisfaction. Also, the most important factors in exercising were physical health, mental relaxation, feeling of pleasure, physical strength.

The results of this study emphasize the need for reinforcement indicators such as mental-emotional, hygienic health, facilities, scheduling programs, in order to maximize the participation of students in sport activities in their leisure time.

The carotenoid-producing yeasts have a widespread distribiution  worldwide. However, isolation of carotenoid-producing yeasts with valuable astaxanthin have been only reported from limited areas and specific habitats. The aim of this study was to apply the multiple polymerase chain reaction method for faster molecular screening of isolates for the yeasts in the exudation sap from birch trees (Betula pendula Roth.) in Alborz Province of Iran with astaxanthin production capacity and toevaluate its efficiency.

This cross-sectional study was carried out by sampling in May 2018 from one of the natural habitats of birch trees Batula Pendula in Alborz Province, Iran. After isolation of yeasts using selected media and purification of yeast colonies with various pink to red colors, yeasts genomic nucleic acids were extracted and molecular screening for astaxanthin-producing detection of isolates was performed by multiple polymerase chain reaction (Multiplex-PCR).

Out of 42 pigmented colonies during sampling and isolation in one cross-sectional study, 23 pink to red colonies were evaluated by Multiplex-PCR method. Of these isolates, three were positive and the rest were negative. The results were validated by determining the ribosomal gene sequencing.

The present study demonstrates the specificity of the multiple polymerase chain     reaction method in fast and cost-effective molecular screening of astaxanthin-producing yeast   isolates and the applicability of this method to study other natural habitats of these yeasts.

Background & A portion of biodiversity and variability of life on earth belongs to the yeasts. The caroptenoid-producing yeasts are economically important and their distinct natural habitats are of interest. In this study, isolation and molecular identification of carotenoid-producing yeasts of the exudates of endangered birch trees (Betula pendula Roth.) at Marmisho in Northwest Iran were performed. Materials & This cross-sectional study carried out by sampling from exudates of birch trees (Betula pendula Roth.) at a natural habitat located in West Azerbaijan Province in the Northwest of Iran. Using selective media, the isolation of the yeasts was done. The screening was initially based on the color and shape of the colonies during one-month incubation along with simple rapid tests to check the existence of carotenoid pigments. Finally, the D1/D2 region rDNA sequencing was accomplished for 19 isolates. 19 out of 45 isolates with pink-red colonies were selected for ribosomal gene sequencing. Phylogenetic results showed that the carotenoid-producing yeasts are in the Basidiomycota division including Rhodotorula, Cystofilobasidium, Cystobasidium, Rhodosporidiobolus, Ustilentyloma, and Xanthophyllomyces.  The unique pattern of carotenogenic yeasts community at this ecological niche and placing in 6 genera with the capability to produce a variety of carotenoid structures indicate that the exudates of birch trees (Betula pendula Roth.) include a wide diversity for native carotenoid-producing yeasts. This study is the first report presenting the diversity of carotenoid-producing yeasts in the Northwest of Iran with a distinct consortium of native yeasts at this habitat.

Evaluation of symbiosis between Arbuscular mycorrhizal fungi (AMF) and Saffron (Crocus Sativus L.) is important because this strategic plant encounters with many environmental stresses such as climatic and edaphic stresses during seasons and the AMF can let the crops increase their productivity along with the improvement of their resistance to stress factors and pathogens
The spores of AMF around rhizosphere of saffron were studied in three fields of Gonabad, Khorasan province, Iran (2013-14). Moreover, the colonization of mycorrhizal fungi with saffron and sorghum trap were studied in three regions using morphologic and molecular methods by nested PCR and amplification of small subunit of rRNA gene fragments.
Three species of arbuscular mycorrhizal fungi, Scutellospora dipurpurescens, Funneliformis caledonius and Rhizophagus aggregatus were identified in the soil around rhizosphere of the saffron of three regions. The colonization of sorghum trap in the soil of saffron cultivation areas was among 21 -41%, while the colonization in the natural Saffron field was 1.5% and just in one area. However, the nested PCR results revealed the colonization of Saffron in all 3 regions. These results showed the colonization of Saffron by Rhizophagus iranicus and F. caledonius.
Discussion and The genus and species diversity of AMF Saffron and Sorghum are different. Moreover, the hereby proposed molecular method is a more precise approach to identify AMF colonization with Saffron while classical methods may provide different and misleading results

The use of accurate and reliable bioassay methods for recombinant proteins is a key element in the production and utilization of such proteins. In this research we have tried to present a reliable method for the G-CSF bioassay. HL-60 cells belonged to human leukemia cells are capable to be differentiated into macrophages or neutrophils. When stimulated by different agents like DMSO, RA and cAMP, HL-60 cells are differentiated into neutrophils, while interferon and phorbol ester differentiate them into macrophages. In this study HL-60 cells were first cultured in flasks. Then 2 x 105 cell/ml of them were selected and 100 ul of this cell suspension was transferred to a 96-well microplate. Then proper dilution of DMSO and RA was prepared and added to the cell suspension (1.3% and 0.1 flM, respectively) and incubated at 37°C and 5% C02 atmosphere. Using RA and DMSO, the cell were transferred from the proliferative phase to the differentiative phase. Then various concentrations of GCSF were added to these treated cells and incubatedat 37°C and 5% C02 atmosphere for two days. The MTT assay was used to measure the reversal rate of the cells from the differentiative to the proliferative phase. The cell proliferation was evaluated by mitochondrial S.T.R. enzme system through the reduction of MTT to Furmazan. The results showed that this method is reliable for the G-CSF bioassay.

Carotenoids are one of the most important and widely distributed pigments in nature and have essential biological functions in organisms. They also impart distinctive and attractive coloration to the breeding animals. Over the past two decades, application of microbial carotenoids has attracted the attention of researchers due to their natural and economic advantages. However, carotenoid producing yeasts with lactose assimilating capacity have rarely been studied and there are few references on the use of lactose as a carbon substrate, such as whey, for the biosynthesis of yeast carotenoids. In this study, a red carotenoid-producing yeast, Rhodotorula acheniorum, was isolated from the sap of birch trees at Masseh Chal in the Taleghan village. The yeast thus isolated was identified through microscopic, macroscopic and biochemical tests. The yeast has both the abilities of lactose assimilation and carotenoid production. The optimal conditions of culture were determined and maximum biomass and carotenoid production were 9.9 g/lit, 290 µg/g, respectively. Chemical analyses of extracted carotenoids have shown that the major portion of the yeast consists of β-carotene, torulene and torularhodin. With regard to the fact that whey is one of the by-products of cheese industries and that its proper disposal has long been a major problem, the most desirable way of handling this waste is to utilize it as a substrate in the production of useful products such as carotenoids.