Molecular evaluation of M2 protein of Iranian avian influenza viruses of H9N2 subtype in order to find mutations of adamantane drug resistance

Background: the H9N2 subtype of influenza A viruses is considered to be widespread in poultry industry. Adamantane is a group of antiviral agents which is effective both in prevention and treatment of influenza A virus infections. These drugs inhibit M2 protein ion channel which has role on viral replication

Objectives: the main objective of this study is to evaluate M gene of avian influenza viruses [AIVs] of H9N2 subtype in order to find adamantane drug resistance mutations

Methods: over 100 suspected samples were collected from different geographical regions of Iran during 2012-2013. Samples were injected via allantoic sac of 9-11 day-old chicken embryos. A total of 11 out of 100 were AIV. The H9N2 subtype was confirmed by specific RT-PCR. The RT-PCR was conducted for full length M gene. PCR amplified products were purified and then conducted for commercial direct sequencing. Finally, sequences were checked for possible sites of adamantane resistance mutations

Results: overall, 8 out of 11 viruses harbored the adamantine resistance-associated mutations. Of which, four viruses were isolated in 2013 and four viruses in 2012. Two different resistance- associated mutations were observed during different years

Conclusions: the present study provided clear evidence concerning resistance AIVs of H9N2 subtype that were circulating in Iranian poultry sector. This concern is always present as M segment might be introduced into human influenza viruses by reassortment phenomenon

Efficacy of ultraviolet radiation on drug susceptibility of Candida Spp. to itraconazole, fluconazole and amphotericin B

Ultraviolet [UV] radiation is an important disinfectant. Fungal infections with resistant isolates in patients culminate in recurrence of disease even with worse condition. This study was done to evaluate the efficacy of ultraviolet radiation on drug susceptibility of Candida Spp. To itraconazole, fluconazole and amphotericin B. This laboratory study was done on 12 Candida spp. isolated from patients according to NCCLS M27- A method. Samples were suspended with sterile saline and optical density was read by spectrophotometer at the wavelength of 530 nm. Serial dilutions [0.0313-16 microg/ml] and [0.0313-128 microg/ml] were supplied for itraconazole, amphotericin and fluconazole, respectively. MICs were determined after 48h incubation at 35°C. Following UV radiation for 1, 2, 5, 10, 60, 90 and 120 seconds MICs were determined, subsequently. The highest MIC pre UV radiation was [>128 microg/ml] for fluconazole. After UV radiation, MICs were steadily decreased for all mentioned drugs while after 10 sec, MICs of itraconazole and amphotericin B were >0.0313 microg/ml. Secondary MICs significantly decreased with respect to MICs obtained in pre UV radiation [P<0.05]. UV radiation reduces MICs of Candida spp. to itraconazole, fluconazole, amphotericin B

Genetic and phylogenetic analysis of the ribonucleoprotein complex genes of H9N2 avian influenza viruses isolated from commercial poultry in Iran

The H9N2 subtype of avian influenza viruses [AIVs] has been isolated in multiple avian species in many European, Asian, African and American countries. Since the first outbreak of H9N2 virus in Iran in 1998, this virus has widely circulated throughout the country, resulting in major economic losses in chicken flocks. Several amino acids in the virus ribonucleoprotein [RNP] complex including the nucleoprotein [NP] and polymerase [PB2, PB1 and PA] proteins are associated with host range and virulence. Our aim was to understand the molecular characterization of RNP complex proteins of Iranian H9N2 subtype isolates. The full length nucleotide sequences of RNP complex genes of two strains designated as Ck/IR/ZMT-101/98 and Ck/IR/EBGV- 88/10 were amplified and sequenced. The phylogenetic analysis revealed that both strains were located in different sub-lineages. However, based on the genetic similarities, PB1, PA and NP genes of Ck/IR/EBGV-88/10 strain had a close relationship with a H7N3 subtype strain, isolated from Pakistan. Most positions of RNP proteins contained amino acids typical of avian determinants of host range. The results showed that the Iranian RNP complex genes have undergone genetic reassortment. Continuous AIV monitoring in poultry industry would help to obtain more information about genetic variation of H9N2 viruses and possible emergence of virulent and/or pandemic viruses

Molecular characterization of non-structural gene of H9N2 subtype of avian influenza viruses isolated from broiler chickens in Iran

The H9N2 subtype of avian influenza viruses [AIVs] have spread in Asia and Middle East countries and have become a serious threat to poultry industry in Iran. Characterization of genes of H9N2 subtype involving in pathogenicity and diagnosis are crucial in control of avian influenza outbreaks. The Nonstructural [NS] gene and its protein products [NS1 and NS2] are important as diagnostic marker, life cycle and pathogenicity of AIVs. The NS gene of five strains, isolated from 1998 to 2010, were completely sequenced and analyzed. All of the examined strains were composed of 890 nucleotides with 230 amino acids. In this regard, only two Iranian strains from GeneBank had 217 amino acids in NS 1 protein. All Iranian H9N2 strains subdivided into two distinct sublineages including I and II. Comparative analysis of NS genes of Iranian strains showed that since 2003, they might have originated from Pakistan H7N3 strains; whereas from 2008 they could be originated from Pakistan H9N2 strains. Although the low pathogenic H9N2 subtype has been permanently circulating from 1998 to the present in Iran, phylogenetic analysis of NS genes revealed that sublineage II has circulated more in poultry industry of Iran. These epidemio-logically variations could be related to vaccination pressure due to massive vaccination or NS gene reassortment in rural and backyard chickens

[Effect of xylanase supplementation on small intestine histomorphometry in broiler chickens]

Exogenous enzymes such as xylanase can cause recovery of efficiency in poultry. In this study we investigated the effects of xylanase supplementation on small intestine histomorphometry in broilers chicken. Two levels of xylanase supplementation [with or without 0.07% xyIanase] was used in 4 treatment groups [two repetitions for each group and 15 chicks per group]. After 42 days, 10 chicks from each pen were euthanized to study the duodenum. jejunum. and ileum histomorphometry. Using light microscope, morphological changes of small intestine including villous height. Number of goblet cells, diameter of epithelium, crypt depth and ratio of crypt depth to villous height were evaluated. Addition of xylanase increased villous height and crypt depth [p <0.05] and decreased goblet cell number and epithelium diameter in different sections of small intestine as compared to the control group. The findings of this study suggest that addition of exogenous xylanase in broiler diets based on wheat can effectively increase the absorption of nutrients

Poultry diseases in Iran: an epidemiological study on different causes of mortality in broilers

Understanding the distributions of poultry diseases will help planning for disease control and prevention more effectively. Studies on poultry diseases in Iran are scarce. We investigated the incidence of mortality in broiler chicken flocks in Iran as part of a national project. Specifically, documents from September 2004 to November 2005 related to the mortality of broilers that were covered by the national insurance scheme were analyzed retrospectively. Of the 439,188,406 broiler chickens that were covered by insurance services, 188,680,459 chickens were exposed to different diseases. The most common diseases [in descending order] were infectious bursal disease [IBD], infectious bronchitis [IB], chronic respiratory disease [CRD], colibacillosis and avian influenza [AI; subtype H9N2]. The mean mortality rate in broilers was 7.89%. Mortality rates were higher during chicks between the third to sixth weeks of age. AI [H9N2] produced the highest mean mortality rate of 26.1%, followed by IB with a mean mortality rate of 22.1%. Most of the mortalities were recorded from the southern provinces of Iran. The distributions of diseases were differed in different regions which could be related to regional conditions and management parameters

Mast cell stabilizers as a potential treatment for Irritable bowel syndrome: a randomized placebo-controlled clinical trial

Mast cells are believed to play a role in irritable bowel syndrome pathogenesis and symptom genesis due to their close neighborhood to gastrointestinal innervations. This study was designed to evaluate the efficacy of orally administered cromolyn for reduction of symptoms in patients with irritable bowel syndrome [IBS]. A randomized placebo-controlled double-blinded 6x6 weeks cross-over study was performed in a private gastrointestinal clinic. 10 patients were allocated to group A and 6 patients to group B. Patients in group A received 150 mg cromolyn divided in three equal doses for the first 6 weeks and placebo for the next 6 weeks but patients in group B received placebo for the first 6 weeks and cromolyn in the next 6 weeks. Weekly evaluation was performed and visual analogue scale was used to determine severity of symptoms. Sixteen patients completed the study. Mean age of the patients was 40.3 +/- 10.9 years old [range: 24-57]. Eight patients had D-IBS [Diarrhea dominant] and other 8 had CIBS [Constipation dominant]. Both cromolyn sodium and the placebo decreased the severity of bloating [Freidman test, p 0.001 and 0.006 respectively]. The severity of the main symptom [diarrhea or constipation] did not decrease in patients of group A and B who were treated with different sequences of the drug or placebo. The severity of pain decreased drastically after 6th week of treatment with cromolyn. Freidman test showed a significant difference between the pain levels of the former defined treatment spots [p 0.01, and 0.02 for patients in group A and B, respectively]. No adverse drug reactions were observed during the study. In conclusion, long term administration of cromolyn seems to be partially effective for treatment of abdominal pain in patients with IBS while main symptoms [diarrhea or constipation] might not decrease during this treatment

Rare peripheral nerve schwannoma

Schwannoma, a benign tumor of nerve sheath origin, is commonly found in the head and neck, as well as flexor surfaces of the extremities. It can rarely occur in the breast and stomach with only a few cases being reported. The cases are: 1- a 57-year- old man with breast mass, 2- a 58 -year -old man with gastrointestinal complaints who were finally diagnosed as peripheral nerve schwannomas. Clinical suspicion and histopathological confirmation are the keys to schwannomas diagnosis. Surgical excision is the treatment of choice for such patients

Non-contrast spiral computed tomography in diagnosis of acute appendicitis

Approach to patients with acute right lower quadrant pain remains a clinical dilemma. Decreasing the risk of negative appendectomies is one of the major goals surgery units intend to achieve. This study has been conducted to determine the accuracy of non-contrast focused appendiceal computed tomography [CT] in preoperative diagnosis of acute appendicitis. During a period of three months, 50 consecutive adult and adolescent patients who were clinically diagnosed as acute appendicitis were included in this study. Focused non-enhanced appendiceal spiral computed tomography [CT] was performed for all patients, preoperatively. Two radiologists who were unaware of the surgical findings assessed the CT scans. After the operation and pathologic assessment, eight patients with negative appendectomy were found. The sensitivity of CT was 0.71 and 0.83 according to the interpretations of the first and second radiologists, respectively. Moreover, its specificity was 0.88 and 0.75 according to the first and second radiologists' reports, respectively. In patients with clinically diagnosed acute appendicitis, relying on abdominal CT is not helpful

Clinical manifestations of irritable bowel syndrome in Iranian men and women: a comparative study

Irritable Bowel Syndrome [IBS] is one of the most prevalent gastrointestinal disorders which includes the majority section of general practitioners or gastroenterologists visits. The type or prevalence of clinical signs and symptoms of the syndrome may be different among males and females. we decided to assess and compare the frequency of different signs and symptoms of IBS in men and women. It is a cross sectional study which was performed in a private gastroenterology clinic in Tehran. 76 males and 116 females who suffered from Irritable Bowel Syndrome were selected, and the clinical signs and symptoms were compared. Finally, the data were analyzed using Chi-Square test. Among the gastrointestinal signs, abdominal pain, nausea, belching, and bitter taste with women/men ratio equal to 1.2, 2, 1.7, and 1.4 respectively [p<.05] were more prevalent in females, while flatulence and mucus discharge were observed in males more than females [men/female ratio 1.3, 1.6 respectively] [p<.05]. Extragastrointestinal manifestations such as palpitation, anxiety, and depression were more common in females [women/men ration 1.7, 1.3, and 1.7 respectively]. The sensitivity of ROME II criteria in males and females was 96.3% and 92% respectively, while the sensitivity of Manning criteria was 93.4% in men and 80.2% in women. Clinical manifestations of Irritable Bowel Syndrome are different between males and females. It seems that Manning criteria is more diagnostic in men suffering IBS

Liver abnormality in celiac disease: report of a case

Many extra-intestinal manifestations are associated with celiac disease. One of the most important of them is hepatobiliary abnormalities. This association is recently under consideration. We report a 34-year-old man with elevated liver enzymes and primary diagnosis of nonalcoholic fatty liver disease. Further follow up revealed celiac disease as the proper diagnosis in this patient

[ accuracy of basic leukocytosis in diagnosis of acute appendicitis]

Background: Some basic laboratory examinations have been suggested as being helpful in the diagnosis of acute appendicitis, including white blood cell count

Materials and methods: In this cross sectional study on adult patients, we decided to assess the accuracy of leukocytosis for prediction of acute appendicitis, in Loghman-Hakim hospital, 2002- 2004

Results: From 736 patients, 456 were male [62%], and 280[38%] were female. 96 patients were observed. The frequency of acute appendicits was 79%. Negative appendectomy rate was 8%. Sensitivity for Leukocytosis with history and physical examination was 84%. The range of specifity was 44-71%. By combining history, physical examination and leukocytosis, sensitivity and specifity increased to 94% and 74%, respectively

Conclusions: Accuracy of history, physical examination and paraclinic combination in diagnosis of acute appendicitis is higher than using one or two of these criteria. However, clinical judgment has the main role in diagnosis of acute appendicitis

accuracy of basic leukocytosis in diagnosis of acute appendicitis

Some basic laboratory examinations have been suggested as being helpful in the diagnosis of acute appendicitis, including white blood cell count. In this cross sectional study on adult patients, we decided to assess the accuracy of leukocytosis for prediction of acute appendicitis, in Loghman-Hakim hospital, 2002- 2004. From 736 patients, 456 were male [62%], and 280 [38%] were female. 96 patients were observed. The frequency of acute appendicits was 79%. Negative appendectomy rate was 8%. Sensitivity for Leukocytosis with history and physical examination was 84%. The range of specifity was 44-71%. By combining history, physical examination and leukocytosis, sensitivity and specifity increased to 94% and 74%, respectively. Accuracy of history, physical examination and paraclinic combination in diagnosis of acute appendicitis is higher than using one or two of these criteria. However, clinical judgment has the main role in diagnosis of acute appendicitis

Nodular regenerative hyperplasia of the liver: report of a case

Nodular regenerative hyperplasia of the liver [NRHL] is characterized by hepatocellular nodules without fibrous septa between the nodules, and has been described in association with certain diseases. The NHRL should be considered in a liver mass and coexisting portal hypertension. We described the case of a 33-year-old Iranian man with NRHL in association with essential thrombocythemia

[ epidemiologic study of microscopic colitis in a university related clinic]

Microscopic colitis has been generally recognized as lymphocytic colitis and collagenous colitis: two inflammatory diseases of large intestine without a definite origin. The colon appears normal by colonoscopy. So, biopsy of the colonic mucosa is mandatory for diagnosis. As a descriptive design we assessed epidemiological, clinical and laboratory findings of patients diagnosed as microscopic colitis from 2001 to 2003, retrospectively. This study had been conducted in one of the University related clinics in Tehran. Results: In this setting, 12 patients were diagnosed as microscopic colitis [3 collagenous colitis, and 9 lymphocytic colitis]. Most of these patients were female [11 patients]. The average of the patients' age at the time of the diagnosis was: 47.6 +/- 18.1 years. The delay of diagnosis was distributed from 2 months to 27 years [average: 8 years]. The most prominent complaint was diarrhea [100%]. Most of the cases were presented as a chronic intermittent manner [6 patients- 50%]. Hyperthyroidism, atrophic gastritis, and type 1 diabetes mellitus, each of them were detected in 3 separate patients. After treatment initiation, recurrence of the disease was not seen during 2 years of follow-up. Most of the time, microscopic colitis is mis-diagnosed as irritable bowel syndrome. Clinical suspicion plays the main role in diagnosis of microscopic colitis cases among patients with watery diarrhea and normal colonoscopy

[Inflammatory bowel diseases and multiple sclerosis: a look on two cases]

The Inflammatory bowel diseases [IBD] have been found to accompany some neurological syndromes including multiple sclerosis [MS]. Recently, Several studies have reported and discussed the simultaneous presence of IBD and MS in some patients. Thus, in patients with either disease, the presence of signs and symptoms of the other should raise the clinical suspicion of the clinician and warrant special considerations. In this report, we introduce two IBD patients who have concomitant involvement of MS

[Clinical course of ulcerative colitis in patients with and without primary sclerosing cholangitis]

We noticed in our practice that the patients with ulcerative colitis [UC] who have developed primary sclerosing cholangitis [PSC] experience a milder course of colonic disease. Our objective in this study was to define whether there is any difference between UC activity and its course in patients with and without PSC. 19 patients with UC and PSC [8 male, mean age 25] were enrolled. To every patient with UC and PSC, 3 patients with UC alone [totally 57 patients, 28 male, mean age 24] matched for age at onset, duration of the disease, and extension of colonic disease were selected as the control group. We used number of hospitalizations due to activity of UC and number of short corticosteroid administrations in various years of follow-up as variables indicating course and severity of the colonic disease in this period. For comparing trends of UC activity between two groups, we used repeated measures two-way analysis of variances. Mean duration of follow up in case and control groups was 12.2 +/- 5.7 and 11.4 +/- 4.9 years, respectively. Two groups had no significant difference in use of sulfasalazine or aminosalicylates. Number of hospitalizations and courses of steroid therapy because of UC activity decreased significantly over time [p<.000] in both groups and it was significantly higher in controls than in cases [p=.045 and .032, respectively]. Development of PSC in patients with UC might have a positive effect on the colonic disease. Further investigation to evaluate basis of this improvement are warranted