[Immunogenic evaluation of recombinant chimeric protein containing EspA -Stx2b- Intimin against E.coli O157 H7]

Background and Aim: Enterohemorrhagic Escherichia coli [EHEC] cause a wide spectrum of infections, such as diarrhea, hemorrhagic colitis and hemolytic-uremic syndrome. Considering the risks associated with antibiotic therapy against EHEC infection, vaccines can be a promising method for prevention of infections. Recombinant chimeric proteins containing multiple immunogens could induce immunity against bacterial infections. The aim of this study was to evaluate immunogenecity of trivalent chimeric antigen EspA -Stx2b - Intimin against E.coli O157 H7 infection

Material and Methods: In this descriptive-laboratory study, recombinant chimeric protein was expressed in E.coli BL21 DE3 by use of IPTG. The protein expression was evaluated by SDS-PAGE and western blotting analysis. The recombinant protein was purified using Ni- NTA affinity chromatography. The immunization was conducted in mice with purified protein and antibody titers were determined by ELISA. Following immunization, mice were infected with E.coli O157:H7 and evaluated for bacterial shedding and mortality. Using SPSS software, statistical analysis was performed by Duncan's test and T-test

Results: The protein was expressed in E.coli BL21 [DE3] and SDS-PAGE analysis showed expression of recombinant protein with molecular weight of 63kD. Western blot analysis confirmed presence of chimeric protein. ELISA results showed that immunogenic properties of chimeric protein induced humoral response to EspA, intimin and Stx2b. Bacterial shedding in immunized mice decreased to 102 cfu/ml and mortality rate was reduced to 60%

Conclusion: The results showed that the chimeric protein induced humoral response and protected the mice against E.coli O157:H7

Serum biochemical and hematological parameters in dogs with benign prostatic hyperplasia [BPH]

Background: clinical prostatic diseases occur in 80% of dogs over 5 and 95% over 9 years of age. It seems that benign prostatic hyperplasia [BPH] affects Scottish terriers more severely than the other breeds

Objectives: this study aimed to evaluate the changes of biochemical and hematological parameters in BPH dogs

Methods: blood samples were collected from 10 male dogs [mostly terrier or mix] older than five years with weight 8.91 +/- 2.5 kg.suffering from BPH which referred to Small Animal Hospital of the Veterinary Faculty of Tehran University. The diagnosis of BPH was based on clinical, laboratory surveys and ultrasonography. 10 normal male dogs with same age, breed and weight were selected as control group. Then serum acid phosphatase [TAP and PAP], CRP, urea, creatinine, total protein, albumin, globulins and hematological parameters were assayed and the results were analyzed by Independent student T-test. Also, Pearson's linear correlation test was used to determine the correlation between TAP, PAP, CRP and ESR with length and width of prostate

Results: the length [p=0.008], width [p= 0.01] of prostates was significantly higher in dogs suffering from BPH compared to the healthy dogs. TAP and PAP levels significantly increased in all dogs in BPH group [approximately 6 times] compared to the controls [p=0.001]. Moreover, serumic CRP concentration was elevated in some BPH dogs [approximately 6 times] [p=0.001]. While there was significant ESR elevation in some of the dogs in disease group compared to the normal dogs, no significant difference was observed in other biochemical and hematological parameters between two groups [p>0.05]. There was a highly significant correlation between serum TAP and PAP [p

Conclusions: the serum acid phosphatase, CRP and ESR were elevated in BPH dogs but the increase in serum acid phosphatase was more important than the others. It is recommended that each laboratory should use its own values of acid phosphatase in dogs

Investigation of MMP-2 and MMP-9 activities in canine sera with dilated cardiomyopathy

Dilated cardiomyopathy [DCM] is accompanied by myocytes and connective tissue changes. Matrix metalloproteinases [MMPs] play important roles in cardiac remodeling. It seems that the gelatinases [MMP-2 and MMP-9] are effective enzymes in cardiomyopathy. Dilated cardiomyopathy was confirmed in 22 dogs [patient group] including 11 female and 11 male by clinical examination, auscultation, thoracic radiography and echocardiography. 17 healthy dogs [control group] with similar weight and breed to patients were also selected from referred cases to Small Animal Hospital of the Veterinary Faculty of Tehran University and the same diagnostic procedures were performed on them. After that, serum MMP-2 and MMP-9 of control and patient groups were measured by semi-quantitative zymography. Semiquantitative analysis of zymograms from canine serums with DCM showed that total MMP-9 in patients is more than control group, while there was no significant difference in total MMP-2 between the two groups. Pro-MMP-2 was not detected in patient group but its active form was present in both groups, of course MMP-2 activity in patients was significantly more than control. Active form of MMP-9 was detected only in patients. Although pro-MMP-9 was present in both groups, its level in control group was significantly higher than patients. The heart enlargement was observed in the left, right or both parts. Statistically significant differences in active form of MMP-2 and MMP-9 levels were observed between different groups of heart enlargement [right, left and both parts] compared to control but this difference was not significant considering chambers affected and VHS [vertebral heart score] groups. In conclusion, although there are some changes in serum MMP-2 and MMP-9 levels in canine DCM, it seems that increase of MMP-9 is more prominent than MMP-2 and neither of them were affected by heart enlargement or VHS grade

Increased serum superoxide dismutase and catalase activities in streptozotocin-induced diabetic rats

One of the diabetes complications is the tissue damage caused by the imbalance of oxidants and antioxidants [oxidative stress]. The aim of the present study was to evaluate the activity of two antioxidant enzymes -superoxide dismutase and catalase- in the serum of streptozotocin-induced diabetic rats. This investigation was conducted on adult male rats assigned to diabetic and control groups. Diabetes was induced by a single intraperitoneal injection of streptozotocin. Seven weeks after diabetes induction, glucose concentration, superoxide dismutase and catalase activities of the serum were assessed. GJucose concentration of streptozotocin-injected animals was significantly higher than that of control group [P<0.001]. The level of Serum superoxide dismutase and catalase activities in diabetes group were significantly higher than those in control group [P<0.01]. There was a positive significant correlation between glucose concentration and superoxide dismutase and catalase activities [P<0.001]. The high activity of antioxidant enzymes in diabetic rats is probably due to compensation responses to oxidative stress produced by high concentration of free radicals. It seems that the higher glucose concentration, the greater compensatory responses

Study of cathepsins involved in haemoglobin and vitellin digestion in Rhipicephalus [Boophilus] annulatus larvae by one- and two dimensional zymography

Enzymatic digestion of proteins in ticks is a complex process and the study of functional proteomics of these enzymes can help to select them as possible vaccine candidates. Blood protein changes [e.g. haemoglobin to vitellin] occur in female mature ticks. Vitellin digestion, as an amino acid and energy source, is one of the vital and important processes in development and evolution of tick eggs and larval stage of unengorged ticks. Several studies reveal a network of proteolytic enzymes involved in haemoglobin and vitellin digestion. These enzymes are mostly cysteine and aspartic peptidases. The aim of this study was the detection of the cathepsins in Rhipicephalus [Boophilus] annulatus larvae extract. In the current research, cysteine proteases extracted from Rhipicephalus [Boophilus] annulatus larvae were studied by one- and two-dimensional zymography. Findings from one dimensional zymography showed a transparent band with 28 KDa. In two-dimensional zymography transparent area are seen in the dark gel background distributed in 21 to 65 KDa zones related to cathepsins. When DTTwas added to incubation buffer [10 mM acetate buffer, pH= 4], the proteolytic activity of some enzymes was increased and appeared as more clear transparent bands in one-dimentional zymography compared with samples incubated in buffer without DTT. As the pH of incubation buffer was acidic and adding DTT resulted in increased activity of the enzymes, therefore, some of these proteolytic enzymes are assumed to be cysteine proteases

Calibration and optimization of a low cost diffusion chamber for passive separated measurements of radon and thoron in soil by Lexan PC SSNTD

Separate radon and thoron measurements in soil are very important in assessment of internal exposure due to inhalation of such radioactive gases. In this study, a low cost, small size, passive diffusion chamber has been developed for simple measurement of Radon-222 and Radon-220[Thoron] gases separately in soil. The diffusion chamber consists of two films and two fiber glass filters. Lexan polycarbonate films were used as Solid State Nuclear Track Detectors [SSNTDs] and optimized film to filter distance was obtained. Calibration factors for the designed diffusion chamber were measured using flow through method which was 16.85[track.cm[-2] [kBq. m-3d] [-1]] and 17.25 [track.cm[-2][kBq.m[-3]d][-1]] for radon for the lower and upper Lexan films, respectively and 1.76 [track.cm[-2] [kBq.m[-3] d][-1]] for thoron. The designed chamber is an economic, applicable and efficient detector for measurement of radon and thoron separately in soil

[Diagnostic value of physical examination for topographic detection of infratentorial lesions in patients with cerebrovascular syndromes]

The Detection of neurologic signs for topographical disorders in central nerves system can prevent unnecessary neuroimaging techniques such as MRI. This study was performed to determine the diagnostic value of physical examination for topographic detection of infratentorial lesions in patients with cerebrovascular syndromes. This descriptive study was done on 200 patients with cerebrovascular syndrome in Qaem Hospital in Mashhad, north-east Iran during 2011. Regarding to topographic physical examination, sensitivity, specificity, positive and negative predictive value and accuracy of signs of syndromes were compared to MRI as gold standard method. The accuracy of tetraparesis and crossed syndromes for localization of brain lesion in brainstem was 79% and 83%, respectively. The accuracy of hemiataxia for localization of brain lesion in ipsilateral cerebellar hemisphere was 98%. The accuracy of Wallenberg syndrome for localization of brain lesion in posterolateral medulla was 98%. Using clinical neuro-examination skills and accurate topographic physical examination can prevent unwanted MRI technique for the diagnoses of cerebrovascular syndromes

Identification of tropomyosin and its immunological properties from larvae of cattle tick, boophilus annulatus

Boophilus annulatus is an obligate blood feeder tick that can cause great losses in animals due to anemia and its ability to injure its host skin directly. The aim of this study was identification of cattle humoral immune response to some tick proteins during experimental infestation. Immune sera against tick were collected from experimentally infested cattle with ticks. One and two-dimensional electrophoresis and Western blotting methods were used for the detection of immunogenic proteins in larval tick extract and eight of these proteins were identified by MALDI-TOF and MALDI-TOF-TOF mass spectrometry. In non-reducing one-dimensional SDS-PAGE, some bounds between 12 to more than 250-kDa appeared. In two-dimensional SDS-PAGE, numerous spot appeared and the identified immunogenic proteins by parallel immunoblotting weighted between 14 and 97 kDa. Amino acid sequences of protein spot with 37-kDa molecular weight had identity to tropomyosin based on Mascot search in NCBI. Anti tropomyosin antibodies can be induced in experimentally infested hosts with ticks and it seems that tropomyosin can be useful for the development of anti tick vaccines.

[Relationship between social support with depression and anxiety during third trimester pregnancy]

This assumption is becoming increasingly accepted within scientific community that variations in fetal environment may have long-lasting over even permanent effects. Considering to alleviator factors of depression and anxiety seems necessary. The Aim of this study was to examine relationship between social support with depression and anxiety during third trimester pregnancy. In correlational study, 95 pregnant women in the city of Shiraz who were selected using several stage cluster sampling in summer of 1387 completed three questionnaires, Social Support, Depression and Anxiety during Pregnancy. To analyze, we used Pearson correlation coefficient and regression analyze. Data analysis showed a negative correlation between social support and depression of third trimester pregnancy [P=0.001], but no significant relationship between social support and anxiety during three trimester pregnancy was found [P=0.09]. Findings of this study show that depression third trimester pregnancy will decrease of increase if social support increase or decrease. This result can be used by the specialists who work with pregnant woman's mental health

Analysis of immunogenic relevant proteins in rhipicephalus [boophilus] annulatus tick

Considering the importance of ticks as a main group transmitting pathogen organisms, this study designed to recognize immunogenic proteins in different tissues of Rhipicephalus [Boophilus] annulatus tick and to find out if there are common proteins in these tissues. Seven cattle were experimentally infested with about 10000 R. annulatus larvae and their humoral immune response to extracts of salivary gland and ovary of adult ticks and larval extracts during infestation were determined by ELISA and Western blot analysis. Measurements of serum antibodies level recorded weekly, from week 0 to week 9. Using Western blot analysis, 15 fractions from soluble antigens extracted from salivary gland and larvae, and 14 fractions in the larval extracts were recognized. These findings illustrate the recognition of common antigens with molecular weight of 170, 117, 100, 70, 37, 33 and 30 kDa from different antigens by resistant cattle sera. Common antigens are present in different tissues of Rhipicephalus [Boophilus] annulatus, which can be used as a target in immunization against ticks

Experimental and Monte Carlo studies on the response of CR-39 detectors to Am-Be neutron spectrum

The Poly-Allyl Diglycol Carbonate [PADC] detector is of particular interest for development of a fast neutron dosimeter. Fast neutrons interact with the constituents of the CR-39 detector and produce H, C and 0 recoils, as well as [n, a] reaction. These neutron- induced charged particles contribute towards the response of CR-39 detectors. Material and Electrochemical etching was used to enlarge track diameter which was made by low energy recoil protons. Before electrochemical etching, a chemical etching was performed for 1 hour. The responses were also calculated by Monte Carlo simulations, using MCNPX code in different energy bins considering H, C and O recoils. The total registered efficiency and partial contributions of the efficiency, due to interactions with each constituent of CR-39, were calculated. The optimized condition of etchant was obtained to be 6N KOH 15kV.cm[1], and 6 hours etching time. The obtained results show that track efficiency of CR-39 was a function of incident neutron energy. The tracks caused by O and C recoil nuclei were negligible for neutron energies lower than 1 MeV. At neutron energies lower than 1 MeV, only recoil protons would have sufficient energy to leave visible tracks. But, O and C recoils had important contributions in overall response of PADC at neutron energies of few MeV. The efficiency of a CR-39 based dosemeter could be calculated by MCNPX code and the results were in a good agreement with experimental results in energy range of [241]Am - Be bare source and [241]Am-Be was softened with a spherical polyethylene moderator of radius of 20 cm

Expression and purification of recombinant mycobacterium tuberculosis [TB] Antigens, ESAT-6, CFP-10 and ESAT-6/CFP-10 and their diagnosis potential for detection of TB patients

One of the most widely used methods to detect tuberculosis [TB] infection is the tuberculin skin test [TST]. The completion of Mycobacterium tuberculosis [M. tuberculosis] genome sequence has led to identification of several antigens that can be utilized for accurate diagnosis and control of TB. The aim of this study was to purify the recombinant M. tuberculosis antigens for the evaluation of their potential in TB diagnosis. The recombinant secretory antigens, ESAT-6, CFP-10 and ESAT-6/CFP-10 were produced by PCR and cloning methods. To investigate antigen specific responses of these recombinant antigens in detection of TB, ex vivo enzyme linked immunospot [ELISPOT] test in 30 clinically diagnosed TB patients was evaluated. The selected M. tuberculosis antigens were cloned, expressed and purified in Escherichia coli [BL21]. ELISPOT assay for detection of TB showed the sensitivity of 93, 90 and 100% for recombinant ESAT-6, CFP-10 and ESAT-6/CFP-10 proteins respectively, which is significantly higher than conventional TST. The recombinant antigens of ESAT-6, CFP-10 and ESAT-6/CFP-10 can be used as an accurate means of detecting TB in Iran

comparative study on the increased radioresistance to lethal doses of gamma rays after exposure to microwave radiation and oral intake of flaxseed oil

Mobile phones, use electromagnetic radiation in the microwave range. On the other hand, there is only one report on radioprotective effects of flaxseed oil. The aim of this study was to investigate the effect of irradiation of rats with microwaves and/or treatment with flaxseed oil on the induction of adaptive response to a subsequent lethal dose [LD] of gamma rays. Eighty male rats were randomly divided into 6 groups of 13-15 animals. The animals in the 1[st] to 5[th] groups received microwave exposure, microwave+flaxseed oil [dissolved in olive oil], flaxseed [continued after LD], flaxseed, and olive oil. At day 5, all animals were whole-body irradiated with a previously reported LD 50/30 of 8 Gy gamma radiation. The 6th group [controls] received the same LD 50/30, but there was not any other treatment before or after the LD. No death event was observed during days 1-9 after LD irradiation in either group. At day 10, death events started in the 4[th] group. Thirty days after irradiation of the animals, the survival fractions for the control group, as expected, was 53.3% while there was no death event in the 1[st] group [survival rate of 100% in microwave-pretreated animals]. The survival fractions for the 2[nd] to 5[th] groups were 69.2%, 92.3%, 46.1%, and 61.5%, respectively. While these findings open new horizons in radiation protection, the radioresistance induced by microwave radiations emitted by a mobile phone may interfere with the outcome of any subsequent therapeutic application of photons or radioisotopes

[Effect of experimental coccidial infection on cellular immunity induced by newcastle disease vaccination in broilers]

In order to evaluate the possible immunosuppressive effects of coccidial infection on Cell Mediated Immunity [CMI] of broiler chickens, 640 Ross male day old broiler chicks were randomly divided into 4 equal groups of 160 [each consist of 4 replicates of 40]. The negative control group remained unchallenged, while the other three groups challenged with 3 different levels of high, medium and low doses of mixed inoculums of E. acervulina and E. maxima at 15 days of age. For the assessment of CMI, Macrophage Migration Inhibition [MIF] test was performed. For this purpose blood samples were collected at 15, 22, 36 days of age. No significant difference was observed among MIF of different groups at 15 days of age [p>0.05]. At 22 and 36 days of age a significant difference observed among MIF of high dose and control groups [p<0.05]. According to the results, it can be concluded that severe coccidial infection may compromise specific CMI activity in broilers

Evolution in treatment methods of hepatic hydatid cyst

A large number of surgical and non-surgical treatment methods have been propounded for cystic echinococcosis of liver so far. For decades, surgical excision via conservative or radical approaches was the only known treatment method for this diseasx Availability of chemotherapeutic agents with considerable activity against Echinococcus granulosus has made it possible to treat hydatid cysts with ultrasound- or computed tomography-guided transhepatic percutaneous drainage method, named Puncture, Aspiration, Injection, and Re-aspiration [PAIR]. The aim of this study was to compare the older and newer treatment methods for liver hydatid cyst. A search was made on three decades later in four different data bases MEDLINE, Chohran Library, Web of Science and Science Direct with keywords: "hepatic hydatid cyst ", "Albendazol", "Cystic echinococcosis", "PAIR", "metaanalysis", Randomized controlled trial Three hundred and seven studies met our inclusion criteria. Most of these studies had considerably large-sized samples of patients with hepatic hydatid cyst. They were comprised of original research articles' case reports, review articles, and randomized controlled trials. These studies had assessed efficacy of conventional surgical methods as well as that of chemotherapy and PAIR. Surgery with different techniques aiming eradication and reductien of recurrence is considered as one the main treatment methods. However, use of percutaneous drainage and minimally invasive techniques is now growing. PAIR with albendazole appears to have higher clinical efficacy, and lower rates of major and minor complications, mortality, short and long term disease recurrence, and fewer days of hospitalization as compared with surgery. Therefore, it is preferable to surgery. Surgery is suggestible in cases for which PAIR is not feasible or complications of the cyst are an issue. Pre- and post-intervention chemotherapy reduces disease recurrence and intraperitoneal seeding of infection

Adhesion and proliferation of horse fibroblast and keratinocyte cells in fibr in glue

Evaluation of autogenous fibrinogen extracted from horse blood in fibrin glue and it's effects on horse's fibroblast and keratinocyte cells were preformed. Fibrinogen has been obtained by glycine precipitation technique from horse's blood. Fibroblast cells were separated by explant culture method from neck skin and keratinocytes were separated by enzyme digestion from lib samples. Autologous fibrinogen, horse`s fibroblast and keratinocyte cells separately mixed with bovine thrombin, calcium gluconate and tranexamic acid. This mixture was uniformly spread on cell culture plate and evaluated for six days. After one day of culture the thickness of fibrin glue were observed by invert microscope, fibroblast cells in elongated shape were observed in the fibrin glue in all thickness and keratinocyte cells were flattened and adherent on surface of plate. Both of them proliferated after 3 days. Fibrinogen that was obtained from horse blood by glycin precipitation technique can support horse fibroblasts as a scaffold and support horse keratinocyte proliferation in vitro

[IOL power calculation in patients with previous RK]

To report the results of different methods for true corneal power and intraocular lens [IOL] power calculation in 10 eyes with previous radial keratotomy [RK] with or without astigmatic keratotomy. In this case series including 10 eyes of 7 patients who had undergone RK with or without astigmatic keratotomy, we firstly determined corneal power using two methods: contact lens method [CLM] and mean keratometry of 3 mm zone in topography. In the next step, IOL power of these eyes was calculated using three formulas including SRK II, SRK T and Holladay II, but the results of Holladay II formula were used as determinant of IOL power during cataract surgery. Since 1.50 diopter change in IOL power results in 1.00 diopter change in patient's refraction at spectacles plane, we estimated the manifest refraction of these eyes with other formulas for comparing with the results of Holladay II formula three months after cataract surgery. Postoperative manifest refraction in 8 eyes by using CLM ranged from -3.00 to +2.00 diopter. In both CLM and mean keratometry of 3 mm zone in topography, the amount of hyperopia after cataract surgery with SRK II formula was more than SRK T, and with SRK T was more than Holladay II. Mean spherical equivalent was 0.08 diopter in mean keratometry of 3 mm zone in topography and Holladay II formula and -0.05 diopter in CLM and Holladay II formula indicating adequate precision of these two methods. It seems that after RK, mean keratometry of 3 mm zone in topography yield a precise estimate of true corneal power as compared to CLM and that Holladay II formula has the most exact result closer to emetropia in comparison with SRK II and SRK T formulas

[Polyclonal antibody production against bovine serum albumin conjugated artemisinin in rabbit]

The aim of the present study was to produce a polyclonal antibody against bovine serum albumin [BSA] conjugated with artemisinin. To gain an immunogenic character of artemisinin, a carboxyl group was added to it using mixed anhydride method. Then, the reactive compound of artemisinin was conjugated with BSA. The BSA+artemisinin were injected to white female New Zealand rabbits for two times. In the first injection, the Fraud's complete adjuvant was used, and two weeks later, a booster injection was carried out with a Fraud's incomplete adjuvant. Two weeks later, blood samples were collected; their serum were separated and frozen until assessment. The production of antibody against BSA+artemisinin was assayed by Immunoblot technique. Antibody was separated and concentrated by saturated ammonium sulfate. The assay confirmed the successful production of an antibody against BSA+artemisinin as a fundamental step in investigation of pharmacodynamics and pharmacokinetics of artemisinin

Etiology and antibacterial resistance of bacterial urinary tract infections in children's medical center, Tehran, Iran

Urinary tract infection [UTI] is a common bacterial illness in children. Knowledge of the antimicrobial resistance patterns of common uropathogens in children according to local epidemiology is essential for providing clinically appropriate, cost effective therapy for UTI. The aim of this study was to determine the distribution of urinary tract infections in a referral hospital, Children's Medical Center, and determination of in vitro susceptibility of these organisms to antimicrobial agents. Of the 1231 bacterial isolates the most frequent isolates were Escherichia coll [38.66%], Klebsiella spp. [22.25%], Coagulase-negative staphylococci [10.1%], Pseudomonas spp. [8.7%], enterococci [8.28%], Enterobacter spp. [4.1%], staphylococcus aureus [3.24%], and proteus mirabilis [2.9%]. Among Enterobacteriaceae, 79.80% of E. coli were amikacin-sensitive. Of Gram-positive cocci, 66.66% of staphylococcus aureus were vancomycin-sensitive. Our data show the original distribution of uropathogens from UTIs in children referred to Children's Medical Center in Tehran and the emergence of multidrug resistant strains

[ evaluation of calulus rate in patients with nephrolithiasis]

Some reports show that oral manifestations such as uremia osteomatiis, self bleeding, periodontal diseases, xerostomia and calculus formation are increased in patients with renal disease. Based on similar structural composition and similar process of production of kidney stones and dental calculus, we performed this study to evaluate the prevalence of dental calculus in patients with kidney diseases. In this case-control study, 30 patients with idiopathic nephrofithiasis were selected as case group and 30 sex and age matched healthy persons were selected as control. They had no systemic and periodontal diseases. Silness-Loe plaque index and Green-Vermillion calculus index were recorded for both groups. Data were analyzed by Chi-square, Fisher's Exact test, T-student and Two-way ANOVA. Results of this study showed that there were not any significant differences between case and control groups in amount of plaque and dental calculus P=0.79. Patients with nephrolithiasis are not more susceptible to calculus formation