Determination of distribution of icsA gene and IcsA protein bands between Shigella flexneri isolated from 3 hospitals in Tehran

Introduction: Shigella is a facultative intracellular pathogen that uses the host actin cytoskeleton protein for intra- and intercellular spread. The aim of this study was to determine the distribution of icsA gene and IcsA expressed protein bands among Shigella flexneri strains isolated from 3 clinical centers in Tehran. Material and Methods: Two hundred and seventy five isolated Shigella flexneri strains were identified by standard microbiological and biochemical methods. DNA isolation was performed using sodium perchlorate method. Hot start-PCR was done with 2 pairs of primers and the products were separated through agarose gel (0.8) in TAE buffer. DNA fragments were visualized by ethidium bromide staining under UV illumination. Whole membrane preparation was used to examine the protein profiles and identification of probable IcsA (120-kda) protein band by SDS-PAGE. Results: From 100 isolated Shigella flexneri strains, both bands of 1600 bp and 1709 bp were detected in 46 isolates (46). A 120 kDa band which seems to be related to IcsA protein was detected in 46 isolates (46). The protein bands varied between 30 and 150 kDa. Discussion: IcsA is both necessary and sufficient for actin assembly in Shigella flexneri. Since icsA gene and IcsA protein band were not found in all Shigella strains, it seems that not all strains have the same pathogenesis. On the other hand, since the demonstration of icsA gene by PCR in all Shigella strains (46) corresponded to the presence of a 120 kDa protein band by SDS-PAGE (46), it seems that both tests may confirm each other. However, the PCR may be more accurate than SDS-PAGE

Immunoreactivity analysis of Toxoplasma gondii recombinant antigen rSAG3 in sera from immunized BALB/c mice and tox-oplasmosis patients

Background: The coccidian protozoa Toxoplasma gondii is an obligate intracellular parasite of humans and other warm-blooded animals. Diagnosis of toxoplasmosis is of considerable medical importance for human, especially pregnant women and immunocompromised individuals. The apply of an Escherichia coli recombinant antigen(s) would be signifi-cantly useful in developing standardization of the diagnostic tests and reducing their costs. In this study, immunoreac-tivity of recombinant SAG3 against sera from immunized mice and human anti-T. gondii IgG positive patients was evaluated by western-blotting and enzyme immunoassay (EIA) in Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences in 2013. Methods: Three inbreed BALB/c female mice were obtained. Two mice were injected with rSAG3 and one was re-mained untreated, as control. Sera from immunized mice and also pooled sera from IgG positive toxoplasmosis cases were evaluated with western-blotting. IgG antibody responses to recombinant SAG3 was measured by indirect ELISA against the negative control group. Results: The rSAG3 protein reacted with sera of immunized mice and sera from patients with anti-Toxoplasma IgG antibodies in western-blot analysis. The result of ELISA showed that, there was marked differences in the absorbance values between the recombinant SAG3 immunized mice and control group. Conclusion: The rSAG3 showed IgG reactivity with sera from immunized mice and anti-Toxoplasma IgG patients. © 2016, Iranian Journal of Public Health. All rights reserved

Combination of gastric atrophy, reflux symptoms and histological subtype indicates two distinct aetiologies of gatric cardia cancer.

<b>INTRODUCTION</b> Atrophic gastritis is a risk factor for non-cardia gastric cancer, and gastro-oesophageal reflux disease (GORD) for oesophageal adenocarcinoma. The role of atrophic gastritis and GORD in the aetiology of adenocarcinoma of the cardia remains unclear. We have investigated the association between adenocarcinoma of the different regions of the upper gastrointestinal tract and atrophic gastritis and GORD symptoms. <b>METHODS</b> 138 patients with upper GI adenocarcinoma and age and sex matched controls were studied. Serum pepsinogen I/II was used as a marker of atrophic gastritis and categorised to five quintiles. History of GORD symptoms, smoking and H.pylori infection was incorporated in logistic regression analysis. Lauren classification of gastric cancer was used to subtype gastric and oesophageal adenocarcinoma. <b>RESULTS</b> Non-cardia cancer was associated with atrophic gastritis but not with GORD symptoms; 55% of these cancers were intestinal subtype. Oesophageal adenocarcinoma was associated with GORD symptoms, but not with atrophic gastritis; 84% were intestinal subtype. Cardia cancer was positively associated with both severe gastric atrophy [OR, 95% CI: 3.92 (1.77 – 8.67)] and with frequent GORD symptoms [OR, 95% CI: 10.08 (2.29 – 44.36)] though the latter was only apparent in the nonatrophic subgroup and in the intestinal subtype. The association of cardia cancer with atrophy was stronger for the diffuse versus intestinal subtype and this was the converse of the association observed with non-cardia cancer. <b>CONCLUSION</b> These findings indicate two distinct aetiologies of cardia cancer, one arising from severe atrophic gastritis and being of intestinal or diffuse subtype similar to non-cardia cancer, and one related to GORD and intestinal in subtype, similar to oesophageal adenocarcinoma. Gastric atrophy, GORD symptoms and histological subtype may distinguish between gastric versus oesophageal origin of cardia cancer

Accuracy of rapid ultrasound in shock (RUSH) exam for diagnosis of shock in critically Ill patients

Background: Rapid ultrasound in shock (RUSH) is the most recent emergency ultrasound protocol, designed to help clinicians better recognize distinctive shock etiologies in a shorter time frame. Objectives: In this study, we evaluated the accuracy of the RUSH protocol, performed by an emergency physician or radiologist, in predicting the type of shock in critical patients. Patients and Methods: An emergency physician or radiologist performed the RUSH protocol for all patients with shock status at the emergency department. All patients were closely followed to determine their final clinical diagnosis. The agreement between the initial impression provided by RUSH and the final diagnosis was investigated by calculating the Kappa index. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of RUSH for diagnosis of each case. Results: We performed RUSH on 77 patients. Kappa index was 0.71 (P Value = 0.000), reflecting acceptable general agreement between initial impression and final diagnosis. For hypovolemic, cardiogenic and obstructive shock, the protocol had an NPV above 97 yet it had a lower PPV. For shock with distributive or mixed etiology, RUSH showed a PPV of 100 but it had low sensitivity. Subgroup analysis showed a similar Kappa index for the emergency physician and radiologist (0.70 and 0.73, respectively) in performing rush. Conclusions: This study highlights the role of the RUSH exam performed by an emergency physician, to make a rapid and reliable diagnosis of shock etiology, especially in order to rule out obstructive, cardiogenic and hypovolemic shock types in initial exam of shock patients. Copyright © 2015, Trauma Monthly

The effects of nanoemulsion cream containing lavender essential oil and licorice extract on healing of deep skin wound in rats

Background: Despite the very good medicinal properties of lavender essential oil and licorice extract, some factors, such as allergic reaction and being biodegradable, limit the application of them as candidates for pharmacotherapeutic treatments. Nano-emulsification is one of the ways to overcome these limitations. The aim of this study was to investigate the effects of nanoemulsion cream containing lavender essential oil and licorice extract on healing of deep skin wound in rat model. Methods: In this experimental study, nanoemulsion cream containing lavender essential oil and licorice extract was made using self-emulsifying method. To investigate its effect on wound healing process, a full-thickness skin wound was produced on 85 male Wistar rats. The wound area was digitally photographed at 2 nd , 5 th , 7 th , 10 th , and 14 th days after surgery using a digital camera; then, the area was quantified using an image analysis system (Image J). Moreover, the expression of transforming growth factor-beta1 (TGF-β1), type I collagen (Col I), and Col III genes was evaluated using real-time polymerase chain reaction (real-time PCR) at days 2, 7, and 14. Findings: Nanoemulsion cream significantly decreased the surface area of the wounds faster than lavender essential oil and licorice extract cream (P < 0.010) and phenytoin (P < 0.050). Moreover, real-time PCR showed that topical treatment of wounds with nanomulsion cream increased the expression of TGF-β1, Col I, and Col III genes in rat�s skin (P < 0.01). Conclusion: Nanoemulsion cream containing lavender essential oil and licorice extract exhibits a promising wound healing potential towards excisional wound models in rats. © 2019, Isfahan University of Medical Sciences(IUMS). All rights reserved

The clinical and environmental spread and diversity of toxigenic Clostridium difficile diarrhea in the region of the Middle East.

Stool samples of 1822 hospitalized patients with nosocomial diarrhea and 100 environmental samples were collected at three teaching hospitals and PCR amplification of rRNA intergenic spacer regions (ISR) was conducted. Bacterial cytotoxicity was assayed by conducting three assays namely toxigenic culture on vero cells, stool cytotoxin, and enzyme immunoassay. ISR was carried out using two universal primers complementary to conserved regions in the 16S and 23S rRNA genes. It was found that the toxigenic culture, stool cytotoxin and enzyme immunoassay showed close rates of detection of toxigenic C. difficile, 124, 121, and 122 /1822 (6.8, 6.64., and 6.7%) respectively. In addition, 32 different ribotypes for toxigenic C. difficile were detected, 28 in clinical and 6 in environmental isolates. The predominant ribotypes from the clinical isolates were 13-15, 35.6%, of isolates. Ribotypes were associated with age, location of isolation, and severity of symptoms of clostridial diarrhea (P<0.05). Ribotypes 6-9 affected children only. The most common ribotype of C. difficile , no. 13, as well as ribotypes 16, 20, and 4 covered almost the whole range of severity of symptoms. Ribotypes 21-27, 1, 3, 6, 7, 9, 11, 14, and 19 caused mild-moderate CDAD symptoms while ribotypes 5, 10 8, 12, 15, 17, and 28 were dominantly of severe symptoms (P<0.05). Environmental isolates showed 17% toxigenic strains composed of 4 different ribotypes while ribotypes 5 was shared with clinical isolates. These findings showed that C. difficile associated with diarrhea were genetically diverse and linked to environmental strains

Human platelet antigen 1-6, 9 and 15 in the Iranian population: An anthropological genetic analysis

Human platelet antigens (HPAs) are membranous glycoproteins considered as alloantigens due to their polymorphisms. HPA-incompatibility in multiple pregnancies or blood transfusion can induce the development of alloantibodies leading to thrombocytopenia. The frequency of HPAs varies among populations, so that deep knowledge of HPA frequencies will help us to reduce those incompatibilities. Herein, we studied the allele and genotype frequencies of HPA1-6, HPA9, and HPA15 among the Iranians with intra- and inter-populations analyses on 36 worldwide populations with diverse ethnicities. The analysis shows that the HPA2 and HPA5 have the greatest differences in genotype distribution between the Iranians and other nations, although similar to other populations, the sole allele found in HPA4, 6, and 9 is �a�. Despite other HPAs, the most frequent allele in HPA15 is �b�, which is also abundant in HPA3. Hierarchical clustering indicates the highest degree of global similarity in HPA genotype frequency among Iranian, Argentinian, Brazilian, and German Turkish populations. Our findings can be applied to decrease the risk of alloimmunizations and platelet disorders, especially in neonates. © 2020, The Author(s)

Artificial spawning and fingerling culturing of European Catfish (Silurus glanis L. 1758)

The present study was conducted to survey the artificial spawning of European catfish (Silurus glanis L.) by synthetic hormones include Carp pituitary extract (CPE), Ovo fact [D-Ala6 des Gly10] GnRHa Ethyl amid in combination with domperidone diluted in propylene glycol, Human gonadotropin chryonic hormone (HCG) and LHRHa (des-Gly10, [D-Ala6] LH-RH Ethyl amide). In this research work 44 male and 72 female brood stokes that transferred to the Shaheed Dr Ansari hatchery and Sefid rod research station were studied on 2 years and injected with those stimulated maturation. Total length, total weight, gonad weight were measured to the nearest 0.1 cm, 1g and 0.01g respectively. The experiments were conducted using different doses of hormones on 4 treatments. In treatment CPE that brood stocks injected with 2.5, 3, 4 and 5 mg/kg/BW, the most responded (83.33%) was with 5mg/kg/Bw of CPE. The mean fertilization in this treatment was 92.5 4.51 and after that there are groups which were stimulated by 4 mg/kg/BW of CPE .In treatment ovafact that brood fish were injected by 10, 20, 30, 40 μg/kg/BW of this stimulation hormone showed 100 percent of fish ovulated in the group treated with 40 μg/kg/BW of ovofact. The mean fertilization rate was 86.08 6.27 in this group. 33.3 % of brood fish ovulated in the group that injected with 40 μg/kg/BW of LHRHa stimulation, but none of brood stock reached to spawning with injection of 30 μg/kg/BW of LHRHa. The brood stocks were injected by 800, 850, 900, 950 IU/kg/BW of HCG, only 33.3 percent of fish in groups that received 900 & 950 IU/kg/BW of this Stimulation hormone reached to spawning stages. According to our results and statistical analysis between trial treatments in percent of Embryo viability showed significant difference (P<0.05) between HCG treatment with Ovofact and CPE groups. This results indicates that embryo viability was better in CPE and Ovofact rate to LHRHa and HCG treatments. In the other cases include the working fecundity, fertilization percent, latency period of maturation, latency period of incubation eggs and ratio of weigh stripped egg to body weight did not show any significant difference (P<0.05) in experimental treatments for stripping the male brood stock of European catfish were injected by 3,4,5 mg/kg/BW of CPE, but not one of male brood fish didn’t reaches to stripping stage , so for eggs fertilization were used of milt from macerated Testes of dead males. So in anther experiment the male brood fish were injected by 5 mg/kg/BW CPE repeatedly at once every 5 days for Period 20 day’s period and milt were collected from of all of the males

Study of artificial production meat possibility in reared beluga (H. huso) with different diets

Four experiments were conducted to study of production meat in reared beluga (Huso huso) with different diets. In the first experiment, A 19-week feeding trial was conducted to evaluate dehulled soybean meal (DHSM) as a fish meal (FM) replacer in juvenile beluga, of initial body weight 8.25±0.08 g (mean±SD) in triplicate groups, fed six isoenergetic (20.1 mJ kg^-1) and isoproteic (45% crude protein) diets, resulting in 5%, 10%, 15%, 20% and 25% of fish meal protein being replaced by soybean protein. Growth performance was reduced significantly with the increasing of DHSM in the diets in the present study. In the next stage, triplicate groups of 315 fish averaging 300.25 ± 10.28 g (mean ± SD) were fed one of seven experimental diets for 14 weeks. Weight gain (WG), specific growth rate (SGR), feed effeciency (FE), protein efficiency of retio (PER) of fish fed 5%, 10% and 15% of soybean protein diets were significantly higher than those of fish fed 20%, 25% and 30% diets. Whole body protein, lipid, moisture content were unaffected among different treatments (P > 0.05). In the second experiment, A 2 × 4 factorial design was used to evaluate the dietary lysine and to determine the optimum dietary L-carnitine in sub-yearling beluga, reared in the indoor system. Twelve experimental diets were formulated and prepared to contain four lysine levels (0.75, 1.5, 2.25 and 4% diet) and two L-carnitine levels (300 and 600 mg/kg diet) at each lysine level. Fish averaging 23 ± 0.5 (mean±SD) were fed one of the experimental diets for 10 weeks. At the end of the experimental period, there were significant lysine and Lcarnitine effects (P 0.05) differences were found in growth performance and feed utilization among the dietary betafine and metionine concentrations in beluga. Hb concentration of fish fed control, 1 and 1.5% metionine were significantly higher than those of fish fed the other diets. FCR of fish was significantly improved by dietary metionine and betafine. FCR was lower (P < 0.05) in fish fed equal ratio of metionine and betafine (2: 2) diets than those in fish fed control diet. Based on above results, it is recommended that the diet for juvenile beluga, Huso huso (8-300g), should contain equal ratio of metionine and betafine (1.5: 1.5) diet, corresponding to 5.95 g/100 g of dietary protein for optimum growth, efficient feed utilization and whole-body protein content. A 17-week feeding trial was carried out to evaluate the effects of dietary L-carnitine level in beluga, Huso huso. A total of fish averaging 1247 ± 15.6 g (mean ± SD) were randomly distributed into 18 fibreglass tanks, and each tank holding 10 fish was then randomly assigned to one of three replicates of six diets with 50, 150, 350, 650, 950 and 1250 mg L-carnitine kg-1 diet. At the end of 17 weeks of feeding trial, average weight gain (WG), feed efficiency (FE), protein efficiency ratio (PER) and condition factor (CF) of fish fed 350 mg kg^-1 diet were significantly (P < 0.05) higher than those of fish fed 50, 150, 950 and 1250 mg kg-1 diets. WG, FE, PER and CF of beluga fed 650 mg kg^-1 diet were also significantly higher than those of fish fed 50, 950 and 1250 mg kg^-1 diets. Whole body and muscle protein were significantly improved by the elevation of dietary L-carnitine level up to 350 mg kg^-1. Liver superoxide dismutase and glutathione peroxidase activities of fish fed 350 and 650 mg kg^ -1 diets were significantly higher than those of fish fed 50, 950 and 1250 mg kg^-1 diets. The dietary Lcarnitine level of 350–650 mg kg^-1 diet could improve growth performance, feed utilization, protein-sparing effects of lipid, antioxidant defence system and reproductive success. Polynomial regression of WG suggested that the optimum dietary L-carnitine level was 480 mg kg^-1 diet. Therefore, these results may indicate that the optimum dietary L-carnitine could be higher than 350 but <650 mg kg^-1 diet in beluga reared in intensive culture conditions

Identification and characterization of pathogenic viruses in aquatic animal using advanced techniques in order to develop rapid diagnostic kit (shrimp viral disease kit as well as TSV, HPV and MBV )

Shrimp production increasing rapidly in the world and in2013 the production reaches 4.2 MT. In Iran the shrimp production is under development and estimated in 1393, 20 thousand tons produced. In this regards the important subject is health and disease in shrimp farm. The white spot syndrome virus for second time appears in chabahar and damage many farms. Because the aquaculture activity expand in the world in national, regional and international scale, many emerge disease are endanger. In this regard the viral disease is very important and not only decrease the production but also has a side effect in business and national economy. For control and prevention the viral disease, the accurate methods such as PCR kit were developed. In this project the PCR methods with sensitivity, specificity and efficacy was designed and used for detection viral disease. Many viruses have several serotypes and in different area maybe new serotype induce the disease. For this reason, the specific kit will be design. Three viruses consist of MBV, TSV and IHHNV are very pathogenic in shrimp farm and need the specific PCR kit for detection them. In this project the MBV virus was identified and designs a new primer with Oligo software and the primer amplified a part of DNA with 185 bp in the gel. The specificity and sensitivity of primer were checked by IQ2000 Kit and the primer used for detection unknown samples