نشریه پلاسما و نشانگرهای زیستی
سال هفتم شماره 4 (پیاپی 28، پاییز 1393)

Inroduction & The first generation of therapeutic monoclonal antibodies was isolated from mouse. One of the disadvantages of them was stimulating immune response in human. Fully human monoclonal antibodies are significantly considered due to their high efficiency and low immunogenic potential. Nowadays different kind of techniques such as phage display and single B cell technology are used to produce fully human mAb. The aim of this project was amplification of VH and VL genes from RNA source of Human plasma cell.With the aim of isolation and amplification of VH and VL regions, single cell RT-PCR reaction was performed. Single plasma cells were lysed by using cell lysis buffer. By using synthesized cDNA from plasma cells and antibody specific primers, antibody genes were amplified. Six pair of primers utilized to amplify the variable region of heavy chain (VH) and light chain (VL). Restriction sites and the linker sequences were placed on primer sequences due to respectively cloned in target plasmid and to link VH and VL. Electrophoresis represented VH and VL fragments with 400 bp length were amplified by PCR. The VH and VL gene sequences were BLAST separately and showed 97% similarity among other antibodies gene sequence. Primer sets were selected and designed which contain linker sequence for ScFv construction, NcoI and NotI restriction sites in order to clone directly into an expression vector.

Inroduction & According to some reports, the Increase in blood glucose level leads to structural and functional changes in reproductive system. In the present study, we attempted to investigate the effect of hydro-alcoholic leaf extract of olive (Olea europaea L.) on the levels of gonadotropin, sex hormones and spermatogenesis in diabetic male rats.48 adult male wistar rats each weighing about 220-250g was randomly divided into six groups of eight. These groups included: control group which left untreated; sham group which received distilled water; experimental group 1, received only 500 mg.kg-1 leaf extract; experimental group 2(diabetic control) receiving only Streptozotocin; and diabetic experimental groups 3and 4 receiving 250 and 500 mg.kg-1 extract respectively. The extract was administered daily by gavageing method for 21 days. Blood samples were taken from all groups and serum levels of LH, FSH, testosterone and Dihydro testosterone were measured by RIA. In this study, the histological changes in testis were cross examined and compared between experimental, control and sham groups.Data were analyzed by SPSS software using ANOVA and Tokay tests.Levels of LH, FSH, testosterone and Dihydro testosterone showed a significant decrease in the diabetic control group in respect to control and sham groups (P-1)exhibited a significant increase, compared to them in experimental group 2(diabetic control). Extract hydro-alcoholic olive leaf, can reduce the side effects of diabetes on levels of gonadotropins, sex hormones, spermatogenesis and can improve pituitary – testis axsis in diabetic rat.

Inroduction & According to some reports, the Increase in blood glucose level leads to structural and functional changes in reproductive system. In the present study, we attempted to investigate the effect of hydro-alcoholic leaf extract of olive (Olea europaea L.) on the levels of gonadotropin, sex hormones and spermatogenesis in diabetic male rats.48 adult male wistar rats each weighing about 220-250g was randomly divided into six groups of eight. These groups included: control group which left untreated; sham group which received distilled water; experimental group 1, received only 500 mg.kg-1 leaf extract; experimental group 2(diabetic control) receiving only Streptozotocin; and diabetic experimental groups 3and 4 receiving 250 and 500 mg.kg-1 extract respectively. The extract was administered daily by gavageing method for 21 days. Blood samples were taken from all groups and serum levels of LH, FSH, testosterone and Dihydro testosterone were measured by RIA. In this study, the histological changes in testis were cross examined and compared between experimental, control and sham groups.Data were analyzed by SPSS software using ANOVA and Tokay tests.Levels of LH, FSH, testosterone and Dihydro testosterone showed a significant decrease in the diabetic control group in respect to control and sham groups (P-1)exhibited a significant increase, compared to them in experimental group 2(diabetic control). Extract hydro-alcoholic olive leaf, can reduce the side effects of diabetes on levels of gonadotropins, sex hormones, spermatogenesis and can improve pituitary – testis axsis in diabetic rat.

Inroduction & A feeding trial was conducted to examine the effects of dietary supplementation of lysine and methionine on growthperformance and haemato-biochemical status in beluga, Huso huso. 240 fish with average weight of 144.6 ± 4.64 g were fed one of four the experimental soy protein based diets D0 (without lysine or methionine supplementation), D1 (lysine supplementation alone), D2 (methionine supplementation alone) and D3 (both lysine and methionine supplementation) were fed to triplicate groups for 12wk. L-Lysine and DL-methionine were added to the diets containing 344 g kg-1 soybean meals at 12.02 and 12.94 g kg-1 of dry diet respectively. Significant higher weight gain, specific growth rate, protein efficiency ratio, condition factor, feed efficiency ratio, haemoglobin concentration, white blood cell and total blood protein were observed in fish fed soya protein-based diet supplemented with both lysine and methionine than that of fish of other dietary groups. There were no significant differences among fish fed diet supplemented with either methionine or lysine. Growth performances, feed utilization and haemato-biochemical status of fish fed control diet was significantly lower than those of fish fed diet supplemented both lysine and methionine and supplemented with either methionine or lysine. The present study showed that soybeen meal could be included in the beluga diet up to 344 g kg1, to replace 40% of fishmeal supplemented both lysine and methionine without any adverce effect on the growth performances and feed utilization.

Inroduction & Parkinson's disease is a degenerative disease in nervous system, which is known with a variety of movement disorders. Factors such as oxidative stress are the main causes of degeneration subestentia nigra dopaminergic neurons and emergence of Parkinsons. Ellagic acid avoids adverse effects of oxidative stress that leads to cell death. The aim of this study is investigate the effect of ellagic acid on movement disorders in animal models of Parkinson's disease. In This study were used 40 adult male rats that were divided randomly into 8 groups: Control, Parkinsons, and three Parkinson groups that once daily for 14 days received the gavage from doses 10, 25 and 50 mg/kg of ellagic acid. Results were presented as mean ± SEM, one-way Anova variance and LSD test. Parkinson's disease was induced with injection of 8 µg of 6_ hydroxy dopamine (6-OHDA) nerve toxin in 2 ml of saline containing 1% ascorbic acid in the middle-anterior bundle (MFB) of the left cerebral hemisphere of rats. lesion by 6-OHDA in the left brain MFB caused up to 14 days after injury animals after subcutaneous administration of apomorphine 2/5 mg/kg, could rotate 360 degrees to the right with rate of> 10 rpm. and in the motor tests (Rotarod performance, catalpsy and Stride length) showed significant differences with compared to the control group. Treatment of animals with Parkinson's disease by different doses of ellagic acid compared with untreated PD group caused a significant improvement in motor activity. doses 25 and 50 mg/kg of ellagic acid showed most effects of treatment.ellagic acid, probably due to strong antioxidant property and interfere with receptor activity in motor pathways can be used to improve and reduced movement disorders, Parkinsons.

Inroduction &

Japanese Threadfin Bream,Nemipterus japonicus, is one of the most important commercial fishes and constitutes an important part of the trawl catch in Persian Gulf. The purpose of this study was Fecundity of Japanese Threadfin Bream, Nemipterus japonicus, in Persian Gulf (the Coastal Waters of Bushehr Province).

In this present a total of 595 specimens collected for absolute and relative fecundity researches. The specimens were caught monthly by bottom trawler in the northern part of Persian Gulf (Bushehr Province).

The fecundity of 41mature were studied, that ranged from 114 to 210 mm in fork length. The mean fork length and gonad weight were calculated 189mm (SD = 27.8) and 3.84g (SD = 2.29), respectively. The maximum and minimum absolute fecundity were calculated 1240910 and 84031related to 210 and 114mm FL, respectively. The mean absolute and relative fecundities were 47238842633 and 3817293 (XSE), respectively. The maximum, minimum and mean of oocyte diameter were 0.448, 0.022 0.221mm (SE = 0.071), respectively.

Inroduction & Today, consume of alcohol is social problems and as well as causing various diseases and abnormalities in physiological disorders in the human body, especially the liver. In this study has tried to, the effect of Lactuca sativa leaf on serum levels of liver enzymes (AST, ALT, ALP, and GGT) and albumin, total protein and liver tissue, followed by alcohol, were investigated. In this study, in order to induce liver damage, ethanol was used. 48 adult male Wistar rats weighing 190 ± 10 g were randomly divided into 6 groups of 8 and with doses of 20%, 40% and 60% of Lactuca sativa and ethanol (30%) were treated. Daily dose of 2 mL of ethanol orally and Lactuca sativa combination with diet was administered daily for 28 days. After blood sampling and separation of serum, the activity level of liver enzymes (AST, ALT, ALP, and GGT) and albumin and total protein in serum were measured and in histopathological studies of liver tissue sections were prepared.Changes in body weight and liver levels in the groups receiving ethanol and Lactuca sativa are not significant compared to the control group. Reducing the level of enzymes (AST, ALT, ALP, and GGT) and increased total protein and albumin in the group receiving Lactuca sativa compared to the group treated with ethanol alone, in the statistics level p ≤ 0.05 was significant. Histological studies showed that group treated with ethanol, there were large areas devoid of hepatocytes. Improvement was observed in the group receiving Lactuca sativa.Lactuca sativaconsumption at a dose of 60% due to higher phenolic compounds and antioxidant activity, cause of protect liver cells is against oxidative stress induced by ethanol, as a result, the level of liver enzymes and biochemical parameters returned to normal and liver tissue damage is minimized.

Inroduction &

Infection with E.coli Bacterium carrying PKS genes is considered one of the causes for infection inside the body, because the bacteria carrying PKS genes will induce Histone H2AX Phosphorylation in antrocites of mice. A considerable increase in frequency of mutations is seen in cells which are exposed to toxin. Studies indicate that infection can cause the potential for mutation. Therefore, colonization of species of E.coli bacteria carrying PKS genes can cause cancer development in large intestine.

In this research, we examined the existence of genetic island of PKS in E.coli bacteria taken from patients’ biopsies infected with inflammatory intestine disease and normal subjects whom have been to Digestion & Glands Clinic at Baghiatalah Hospital and the contrast between cancerous and healthy tissue was performed using a scanning electron microscope. Therefore, 20 biopsy samples of healthy people and 25 biopsy samples of intestine tissue from people infected with inflammatory intestine disease were examined in terms of microbiology and molecular structure. For this purpose, E.coli bacteria was detected by using the intestine tissue, microbiologic and biochemical approaches, then the their DNAs was extracted, and Multiplex PCR approach was used for the specific designed primers in terms of availability of genes in the PKS island, and the PCR products was examined using Electrophoresis on 2% Agarose Gel.

The results have indicated that 28% of the E.coli bacteria taken from patients’ biopsies have genetic area of PKS.

Introduction & Objectiv:Diabetes Mellitus is one of the most common endocrine diseases and currently affects 382 million people. One of the problems with diabetes causes skin ulcers leading to amputation. Balancing the indigenous skin micro flora and reducing skin opportunistic pathogen play an effective role on preventing or reducing skin scars. Prebiotics are used in various industries to increase the resistance against disease and health promotion. The objective is to study the prebiotic effect of Saccharomyces cerevisiae AT-50 cell wall to inhibit the growth of gram+ive, catalase+ive cocci isolated from normal skin between the toes in diabetics. It aims to design a method for prevention of diabetic ulcers.Healthy skin between the toes in 53 diabetic patients and 50 non- diabetic patients were sampled.Gram +ive, catalase -ive strains were isolated. Saccharomyces cerevisiae AT-50 cell wall prepared by sonicator, followed by evaluating prebiotic effect of 1.5 and 3 percent of the pure cell wall on coagulase +ive isolates using well method. The inhibition zone diameter were measured in millimeters and compared as inhibitory effects.The results showed that the total count ratio of anaerobic and aerobic isolates from diabetic and non diabetic samples was 1.44 and 1.24, and the total count ratio of aerobic and anaerobic isolates from diabetic samples were 1.55 and 1.81, respectively.Also, the number of coagulase +ive gram +ive cocci isolated from skin samples from diabetes, was 75 % more than the control sample.The skin of people with diabetes, regardless of age and sex to non- diabetic healthy skin had more favorable conditions for microbial growth, especially for coagulase +ive cocci. Inhibitory effect of prebiotic Saccharomyces cerevisiae AT-50 cell wall was related to dosage. Inherent resistance was seen in diabetic patients. These results can be used in the future as a model for the prevention, reduction and control of chronic wounds such as diabetic ulcers.