فصلنامه تازه های بیوتکنولوژی سلولی مولکولی
پیاپی 30 (بهار 1397)

Aim and Today, the presence of broad-spectrum beta-lactamases in bacteria isolated from patients who express multiple drug resistance is a major health problem in most countries.
A total of 346 samples of different clinical strains of enterobacteriaceae were isolated from Kahooresten. To determine the resistance of the samples, an antibiotic test was performed. Finally, ESBL isolates were tested by PCR for TEM, CTX-M, and SHV genes.
Of the 346 isolates studied, 162 isolates of Klebsiella and 132 isolates of Escherichia coli were resistant to ceftazidime antibotic or cefotaxime, or both. Also, 82 (42.26%) were isolates producing ESBL.
Due to the high percentage of resistance to third-generation cephalosporins, accurate antibiotic testing before antibiotics is required in infections caused by ESBL-producing organisms.

Aim and Because of high half- life of niosomes in blood plasma makes the possibility of delivery of chemotherapy agent to tumor tissue and improves the therapeutic index of chemotherapy drug. In this study, the nano-nisomes contained doxorubicin was synthesized. The effective dose of drug was determined and anti-cancer effect of resulting nanoparticle was evaluated.
Material & Niosome containing cholesterol, span 60 with the 85:15 molar ratios were prepared by thin film hydration method. In the hydration step, various concentration of doxorubicin diluted with distillated water were used to determine the optimized drug concentration. The cellular cytotoxicity was finally examined using MTT assay.
The results imply that optimized drug dose is 0.5mg/ml. The entrapment efficiency; size diameter and polydispersity index of optimal formulation are 81.69, 102.9 and 0.128, respectively. The amount of drug release is 35% during 144 hours. The prepared system reduces the side effects of doxorubicin and be effective against cancer cells.
This study showed that the optimal dose of drug plays an important role in improving the percent of drug loading that is economically optimal. Also it leads to expose the patient's body with the lower doses of medication and the most therapeutic effect. Reduce the dose of medication also causes less damage to healthy cells. Sustained-release property of system is the main reason for the increased toxicity of chemotherapy drug.
Prepared niosomal system is slow release with size diameter less than 150 nm and high drug entrapment efficacy that can be used to overcome the side effect of free doxorubicin. The resulting system is effective against bone marrow cancerous cells.

Aim and Extended-spectrum beta-lactamase of CTX-M type is considered as an Important mechanism resistant to beta-lactamases in gram- negative pathogen and is widely increasing. Klebsiella pneumoniae species are able to produce extended-spectrum beta- lactamase (ESBLs). The aim of this study was to detect the prevalence of CTX-M gene in ESBLs produsing Klebsiella pneumoniae isolated form patients in different region of TEHRAN city from December2016 to july 2017 by using PCR method.
Material and In this analytical- descriptive study antibacterial susceptibility patterns of 176 Klebsiella pneumoniae tested to Cefepime, Ampicillin, Gentamicin, Amikacin, Cefalotine, Ceftazidime, Piperacillin, Ciprofloxacin, Imipenem, Cefotaxime , Nitrofurantoin, Trimethoprim using disk diffusion method. In addition, confirmatory test for detecting ESBLs phenotypes was performed using Cefotaxim-Clavulanic acid combination disk. The presence of CTX-M gene was assessed by using PCR.
Confirmatory phenotypic test showed 56.81% of the isolates were ESBL positive. The prevalence of CTX-M gene in isolated Klebsiella pneumoniae was 28%. In this study, the highest resistance was observed for ampicillin antibiotics (24.88%) and the least resistance to antibiotic imipenem (6.06).
High frequency of CTX-M gene in ESBL produsing isolates indicates that this enzyms plays an important role in resistance to beta lactam containing anti biotics. Therefore, proper infection control tools and appropriate therapeutic approaches in different parts of the hospitals are necessary to prevent their release.

Aim and Ducrosia anethifolia Boiss is from the Apiaceae family, which is used in traditional medicine as a headache and back pain remedy. This research was conducted to select the best pretreatment for breaking seed and seed germination and seedling growth.
Material and Experiment was conducted in a completely randomized design with 6 treatments and 4 replications in University of Kashan in 2016. Treatments included potassium nitrate 0.1% and 0.2% for 72 hours, sulfuric acid 98% for 5 minutes, chilling for 35 days at 5 ° C, soaking in distilled water and control.
The results showed that the effect of different treatments on germination percentage, germination rate, root and stem length and fresh and dry weight were significant at 1% probability level. Sulfuric acid scarification treatment completely prevented germination.
The highest germination percentage was observed in potassium nitrate solution. Therefore, potassium nitrate is useful in response to metabolic processes of the seeds
Considering that the most important reason for reducing the distribution of this plant is the length of the seed dormancy, the use of the results of this study will help to germination of this herb.

Aim and Abortion is the most common complication in the first and second trimester of pregnancy. The concept of abortion refers to the termination of pregnancy before the twentieth week of gestation. Women who experience consecutive abortions more than twice can be said to be suffering repeated abortions.
This study aims at investigating the relationship between the XPO5 rs11077 polymorphism and DROSHA rs10719 miRNA machinery gene in women suffering spontaneous repeated abortions so that it can be used as a diagnostic marker for women with repeated abortions if such a relationship is confirmed.
The blood samples of 50 women with spontaneous recurrent abortions and those of 24 women with no history of abortion and at least two successful births were used in this study and the women were selected from those admitted to Masoud labaratory in Tehran. DNA was extracted using a special DNA extraction kit from whole blood samples with EDTA. The XPO5 rs11077 gene polymorphism 2 and DROSHA rs10719 were investigated using the Tetra-ARMS PCR method and analyzed using agarose gel electrophoresis.
The frequencies of Genotypic and allelic rs11077 (A / C) polymorphism of XPO5 gene did not show any significant difference between the two patient and control groups. However, the frequency of genotypic and allelic rs10719 (T / C) polymorphism of DROSHA gene showed a significant difference between the patients and the control samples and TT genotype was shown to be significantly higher in the patient group than in the control group.
Our findings suggest that the rs119 (T / C) polymorphism of the DROSHA gene may be associated with the susceptibility to RPL in the Iranian population, and the T allele may increase the incidence of this disease.

Aim and One of the most important researches is related to garlic and it has great anti-bacterial properties. The purpose of this research is the evaluation of garlic extract effect with various concentrations on Staphylococcus aureus, Streptococcus pneumonia and Shigella dysenteriae bacteria and also comparing the results in laboratory situation.
Material and In this study, garlic extract was prepared and to determine the Minimum Inhibitory Concentration (MIC) of the extract, the Standard Tube Test was used. As well as, to compare with antibiotics, the Disk Diffusion Method was used. The experiment was conducted in a completely randomized design with 3 repetitions and comparisons of the averages were performed using Tukey's method.
The MIC of garlic extract based on Mg/dl on studied bacteria with extract of respectively: 15, 23 and 25mm with concentration of 1/2 Mg/dl; 12, 20 and 22mm with concentration of 1/4Mg/dl; 10, 16 and 18mm with concentration of 1/8 Mg/dl; 0, 13 and 15mm with the concentration of 1/16, 1/32 and 1/24 was zero. According to the results of this research in laboratory condition anti-bacterial properties of garlic has the inhibitory effect on studied bacteria.
The results indicate that the inhibitory effect of garlic extract on studied bacteria, in laboratory environment, is as good as antibiotics.

Aim and This study was performed to determine the evolutionary and molecular trend of BLG gene nucleotide sequence in mammals, phylogenetic analysis and the trend of natural selection.
The nucleotide sequence of the BLG gene in studied species were obtained and balanced from genome database (NCBI). Substitution and replacement percent of nucleotides were obtained using maximum likelihood, and phylogenetic tree drew using Neighbor-Joining method.
The results showed that the transitional substitution was more than transversional substitution. The numerical value of dN/dS was 0.70, indicating negative selection during evolution of this gene. The results of phylogenetic tree for this gene showed different evolutionary routes in different species including five categories such as (cow and buffalo), (sheep and goat), horse and wild boar, in which cow, buffalo and sheep had the most relationship.
Mutations and natural selection have been resulted in the development of new varieties, new proteins and also stabilizing their performance during the evolution and progress toward purification of their performance.
According to the results of this study, protected areas constitute a small part of LGB gene sequence which reflects the polymorphism of this gene as well as being susceptible to variations and mutations.

Aim and Due to the high prevalence of diseases caused by the presence of lead in soil contaminated with petroleum industrial centers, then transfer the metal to plants and animals and eventually humans and high risks for human health and other living things. It seems that identify and isolate microorganisms with high strength steel and the biological removal can be an important step in the evolution of contaminated soil cleanup systems considered industrial centers.
Material and The was sampling of three areas of oil contaminated soils C-branch Masjed Soleiman. To isolate resistant bacteria lead ΙΙ PHG agar dilution method and the culture medium containing concentrations of 0.5, 1, 2, 2.5, 3, 3.5 and 4 mMol per liter of lead were used. To identify of resistant bacteria lead , first was used of biochemical tests and then molecular methods PCR- colony.
In this study, 5 resistant bacteria lead from oil contaminated soils were isolated and identified that among of them Proteus mirabilis strain AOUC-001 bacteria and showed maximum resistance to lead (MIC=4mM).
Discussion and According to the bacteria examined in this study could lead bioabsorption in -vitro and growth rather appropriate in the presence of the metal showed, so it seems a good candida for bioremedation of lead from oil contaminated soils in the future. Provided that the contaminated soils to isolate the bacteria to be filtered out treatment.

Aim and Urinary tract infections (UTI) are considered as the most common bacterial infections in young children, adults and women. Therefore appropriate treatment is necessary immediately after diagnosis. In the present study strains of Group B Streptococcus were isolated from urine to determine the drug-resistant.
This cross-sectional descriptive study was carried out on 3676 patients at reference laboratory of Salmas city in 1394 of which 2754 were women and 922 were men. Mid-stream urine samples taken from patients were transferred to the Todd Hewith media then were cultured in blood agar. Antibiotic susceptibility of streptococcal strains and drug-resistance patterns were conducted using the disk diffusion method on the Mueller Hinton culture medium according to guideline standards.
Out of 3678 samples, 47 samples (36 females and 11 males) showed positive results. Group B Streptococcus was mostly resistant against oxazoline and clindamycin. The rest of drug-resistancy pattern respectively includes gentamicin, tetracycline, ceftizoxime, amikacin, cephalexin, trimethoprim, ampicillin, ciprofloxacin, and cefixime.
Considering the prevalence of urinary tract infections, especially in children under 2 years and also in girls, the knowledge of local resistance pattern and well-timed treatment are imperative. This study showed that group B Streptococcus represents the highest frequency of drug resistance. This issue requires more attention of Health officials is Salmas city.

Aim and Urinary tract infections is one of the most common infectious diseases whichmany factors are involved, but bacteria such as E.coli is the most important agent of urinary tract infections. Antibiotic resistance as a major problem in the treatment and control of these infections is considered. The aim of this study was to determine the genes that cause resistance to betalactam family of antibiotics on E.coli isolated from urinary tract infections in Robat karim city.
Material & In the present Cross-sectional study which was conducted over a period of 6 months, 123 samples of E.coli were collected from Robat karim hospitals for molecular analysis of TEM genes, causing antibiotic resistance by (PCR) method.Data were analyzed using SPSS statistical test.
PCR showed that the gene frequency of TEM (80%) and the highest and lowest prevalent of genes were related toTEM in E.coli isolated from urinary tract infections respectively.
According to the high prevalence of resistance to beta-lactam antibiotics, the current study showed that the noted geneTEM play an important role in facilitating the spread of antimicrobial resistance in this region

Aim and Lincomycin is a constituent of the lincosamide group, which typically feature a 6, 8-dideoxy-6-aminooctose lincosamine. The purpose of this study was to optimize production of lincomycin by streptomyces lincolnesis PTCC 1629 using classical one-factor-at-a-time method.
Material and Streptomyces lincolnesis PTCC 1629 was provided from Persian Type Culture Collection, Tehran, Iran. Starch casein agar and yeast extract-malt extract media were used for primary culture and production of lincomycin, respectively. Optimization of carbon and nitrogen sources were performed according to classical one-factor-at-a-time method. Antimicrobial susceptibility tests were analyzed using standard strains including; Micrococcus luteus PTCC 1408, Staphylococcus aureus PTCC 1189, Streptococcus pyogenes PTCC 1447, Staphylococcus epidermidis PTCC 1435, Bacillus cereus PTCC 1154, Escherichia coli PTCC 1395, Salmonellatyphi PTCC 1609 and Shigella flexneri PTCC 1234.
The obtained results showed that glucose and potassium nitrate were the most suitable carbon and nitrogen sources for lincomycin production by S. lincolnesis PTCC 1629, respectively. Amongst the test strains gram positive bacteria were susceptible to lincomycin production by S. lincolnesis PTCC 1629, but gram negative test strains were resistant to this antibiotic.
Theobtainedresults in this study showed that optimization of carbon and nitrogen sources could improve lincomycin production by S. lincolnesis PTCC 1629 with significant antibacterial activity against gram positive bacteria.

Aim and Male urogenital tract infection is one of the most causes of male infertility. Among bacterial species, E.coli is the most important pathogen thatcaused defection in spermatogenesis. The purpose of this study was todetermine of fim H,pap and afa genes in E.coli isolated of infertile men semen.
100 semen samples were cultured. Bacteria were identified according to standard bacteriological methods. Then DNA was extracted from E.coli isolated and frequency of afa,pap and fim H genes were analyzed by PCR.
from 100 samples of semen,53 bacteria were isolated, as, E.coli (18.86%), staphylococci (33.33%), streptococci (45/09%) and klebsiella (5/88%). E.coli infected samples had abnormality in sperm motility and morphology. Frequency of fim H and pap genes in E.coli strains were 100%, also afa gene didn`t find in any of samples.
results showed that 18.86% E.coli was isolated from infertile men semens. E. coli is an important genital pathogen and prevalence of the pathogenic genes, including adhesions, is effective in the control and treatment of the male infertility. This study indicated the prevalence of fim H and pap genes in E.coli contaminated samples could be related to males infertility.