Detection of InvA Gene in Non typhoidal salmonellae isolated from food products and clinical cases

Salmonellae are well-known pathogens, highly adaptive and potentially pathogenic for humans and/or animals. Salmonellae are capable of producing serious infections that are often food borne and present as gastroenteritis. The main reservoirs for non-typhoidal Salmonella are animals such as poultry, livestock, pets and reptiles. Salmonella typhi and Salmonella paratyphi colonize only in humans, so they can be acquired only from close contact with a person who has typhoid fever, from a chronic carrier, or from water or food contaminated by human feces. Determine the virulence of the Salmonella serovars obtained from human and animal sources by investigating the presence of chromosomal virulence gene, invA gene as it triggers the internalization of the organism required for invasion of deeper tissue. A total 480 clinical samples: 120 milk samples were collected from faculty of agriculture farm and Markets, 115 eggs were received from different farms and markets, 125 fresh slaughtered chicken and 120 stool sample were collected from Assiut Children University Hospital. Invasion gene operon, invA was detected in all 50 clinical isolates representing multiple serotypes from different sources. Identification of S. enterica and screening of invA gene through PCR based procedures can have major benefit in public health specifically for rapid diagnosis, epidemiological investigations, ideal vaccine, development of treatment, and prophylactic strategies for salmonellosis

Detection of non typhoidal salmonella isolated from food products and clinical cases by PCR and conventional methods: a comparative study

Non typhoidal Salmonella [NTS] are important food-borne pathogens. Infection with NTS may not lead to fatal disease, hut it may remain localized in the gastrointestinal tract resulting in gastroenteritis or may take a septicemic form that can affect several organs systems causing gastroenteritis, bacteremia and subsequent focal infection. To compare PCR with different conventional methods for identification of non-typhoidal Salmonella species, and to determine the virulence of the Salmonella serovars obtained from human and animal sources by investigating the presence of virulence gene, InvA in the chromosomal DNA. A total of 480 clinical samples were collected. These included: 120 milk, 115 eggs and 125 fresh slaughtered chicken from farms, slaughterhouses, markets, in addition to 120 stool samples from Assiut Children University Hospital. They were subjected to conventional methods for bacteriological and biochemical examination. Conventional cultural examination, API 20 E system and PCR amplification assay of virulence gene invA were investigated in animal and human isolates. By comparing the results of PCR using SI 39 and SI 41 primers and those of culture examination, it was found that PCR had similar results to culture examination. PCR could detect 50 positive cultures of Salmonella species, while API 20 E could detect only 47 of these positive cultures.: PCR amplification assay has the ability to detect a wide range of Salmonella species depending on the design of primers targeted to invasion gene operon [InvA gene] of salmonella. PCR technique may provide a valuable, rapid, specific and sensitive laboratory diagnostic test for detection of salmonella DNA in cultures

Prevalence of transfusion transmitted virus [TTV] infection in patients with liver diseases attending to Assiut University Hospital

In 1997 Transfusion Transmitted Virus [TTV] was isolated from the serum of a patient with post transfusion hepatitis of unknown etiology, in Japan. It's considered as a causative agent of non A to G hepatitis. To assess the prevalence of TTV infection among patients with liver diseases compared with healthy controls and the significance of TTV infection in patients with liver disease. This investigation was conducted on 76 patients with liver diseases, classified into four groups: Acute hepatitis group [20 patients], chronic liver diseases [30 patients], Liver cirrhosis [18 patients] and hepatocellular carcinoma [8 patients]. In addition to the patient groups, the fifth group of 24 healthy blood donors as control group was included within the study. All patients and control groups were examined for the detection of TTV DNA by PCR. Thirty seven had history of blood transfusion and 23 patients were subjected to surgical manipulation. TTV DNA was detected in 57.9% [44/76] of patients with liver diseases and in 45.7% [11/24] of healthy blood donors. The prevalence of TTV in the studied groups were 60%, 46.7%, 66,7% and 75% in acute hepatitis, chronic liver diseases, liver cirrhosis and hepatocellular carcinoma respectively. TTV is commonly present in patients with liver disease attended to Assiut University Hospitals as well as in blood donors. High prevalence of TTV in blood donors may indicate other routes of transfusion of this virus such as fecal-oral and sexual routes beside transfusion of blood and blood products. The blood transfusion and operative intervention are a major risk factor for transmission of TTV

study of T-helper 2 / T-Helper 1 imbalance in respiratory syncytial virus bronchiolitis in relation to severity and outcome

The present study was carried out on 80 infants with acute bronchiolitis. Their ages ranged from 2-22 mo. They were 60 males and 20 females. 30 apparently healthy infants of matchable age and sex were enrolled in the study as controls. Besides taking history, thorough clinical examination and routine laboratorial investigations, arterial oxygen saturation, RSV tissue culture of nasopharyngecil lavage, direct immunoflourescence for serum RSV Ig-M, serum IL-4 and INF-gamma were done. Results showed that viral bronchiolitis is predominant in males around 6 mo of age and in winter months. RSV infection was found, in 72.5% of cases. Serum IL-4 was not different between RSV negative patients and controls but it was significantly higher in RSV positive patients than either RSV-ve cases or controls [P< 0.001 for each]. Furthermore in RSV + ve patients it was significantly higher in severe and fatal cases than in milder and surviving ones [P<0.001 for each]. Serum INF-gamma showed no significant difference between RSV + ve and RSV -ve patients hut it was significantly higher in both groups than controls [P< 0.001 for each]. In RSV+ve cases it was significantly lower in more severe and fatal cases than milder and living ones [P<0.05 respect]. The ratio I-L-4/ INF-gamma was significantly higher in RSV + ve group than RSV-ve and than controls [P<0.001 and P< 0. 05 respectively]. ARC and SaO[2]% showed significantly lower values in RSV + ve patients than RSV-ve ones [P<0.001], but the latter showed significantly higher AEC than controls [P < 0.05]. In RSV + ve group, both parameters were significantly lower in severer and fatal cases than milder and living ones [P < 0.001 for each]. In RSV positive group, significant positive correlations were found between disease severity and either serum IL-4 or IL-4 /INF-gamma ratio [r=0.789 and r=0.823, P < 0.001 respectively] but disease severity was inversely correlated with either AEC or SaO2% [r = - 0.962 and r = - 0.828, P < 0.001 respectively]. RSV is the major etiologic virus of bronchiolitis in young infants. Viral tissue cultures and direct immunoflourescene are of equally diagnostic value but both may be needed. Most of the severe and fatal cases are in RSV + ve group. Th2 / Th-l imbalance reflected on IL-4 /INF gamma ratio is more deranged in RSV bronchiolitis with Th-2 predominance especially in more severe and fatal cases. These conclusions may be a rationale for addition of antiviral in severe cases of bronchiolitis but further studies of antiviral drugs and their effect on cytokines may be needed to address this recommendation

Seroepidemiological study on helicobacter pylori infection in children and adults in Assiut Governorate, Upper Egypt

Helicobacter pylori is considered as one of the most common chronic bacterial zoonotic infections worldwide. H. pylori infection constitutes a public health hazard problem with a fatal sequele. This study was designed to estimate the prevalence of H. pylori among children and adults in Assiut Governorate in Upper Egypt and to evaluate the risk factors affecting the increase of infection. A total of 200 peripheral blood samples constituting 50 samples from children and 150 samples from adults were collected from hospital patients. A full detailed anamnestic and clinical assessment in the form of questionnaire was designed for each patient to determine the risk factors with specific emphasis to age, sex and residence. Seroprevalence of H. pylori IgG antibodies was evaluated by using enzyme linked immunosorbent assay. We estimated a seroprevalence rate of 75.5%. H. pylori exposure was higher in adults [79.33%] than in children [64%] and it was statistically significant. The seroprevalence of infection was higher in rural areas [87.3%] than in urban areas [40%]. H.pylori infection was prevalent among humans in the areas studied. Public health hazard of H. pylori infection and preventive measures to control the infection were discussed

Demonstration of peripheral benzodiazepine receptor-subtypes in the rat phrenic nerve diaphragm preparations using flumazenil

The effect of graded cumulative concentrations of the benzodiazepine [bd] receptor agonists, diazepam and midazolam, on the height of diaphragmatic twitches of rat phrenic nerve diaphragm preparations was investigated both in the absence and presence of the central bd receptor antagonist, flumazenil. The statistical analysis of the results demonstrated that the two bd agonists investigated in equimolar concentrations ranging 0.02-0.32 mM had different effects on twitch tension. Diazepam elicited a biphasic response consisting of an increase of twitch tension by 14% and 24.5% at a lower concentration [0.02 and 0.04 mM, respectively] and an inhibition by 21%, 60% and 96% at higher concentrations [0.08, 0.16 and 0.32 mM, respectively]. On the other hand, midazolam induced an inhibition of twitch tension by 14%, 20%, 53%, 75% and 99% throughout the whole range of the tested concentrations with no initial stimulatory effect on twitch tensions. Comparing the inhibitory concentration 50 [IC50] of diazepam with midazolam, midazolam was found to be twice potent in eliciting its inhibitory effect [IC50, 0.07 mM versus 0.14 mM for diazepam]. The central bd receptor antagonist, flumazenil [0.1 mM], specifically antagonized the potentiating effect of diazepam on twitch tension, but failed to antagonize the inhibitory effects of both diazepam and midazolam

Aminophylline improves contractility of the isolated rat phrenic nerve diaphragm preparation study of the possible mechanisms

Lately, the role of aminophylline in improving diaghragmatic contractility in respiratory outflow obstruction, was emphasized as a principal mechanism outweighing its bronchodilator effect. In the present work, using the isolated rat phrenic nerve diaphragm preparation, the effect of aminophylline on contractility and the possible mechanisms involved were investigated. The effect of aminophylline applied cumulatively in doses of 0.25, 0.5, 1, 2, 4 and 8 mM was tested on fresh nerve stimulated preparations and compared with its effects on directly stimulated diaphragm muscles, muscles treated with verapamil and fatigued muscles. Aminophylline induced a statistically significant increase in the basal height of the diaphragmatic twitches in a dose dependent manner reaching a maximum of 128.5% in the fresh nerve stimulated muscle, 127.8% in the directly stimulated, 88.4% in the verapamil treated; and 305.2% in fatigued muscles. Moreover, partial and complete blockade of adenosine receptors in the phrenic nerve diaphragm preparations by 25 and 50 UM aminophylline respectively, were also demonstrated. These findings indicate that aminophylline improves diaphragmatic contractility by a direct effect on the muscle most probably through increased Ca [++] influx, an effect that might be mediated through blockade of adenosine receptors. The improvement in diaphragmatic contractility was more pronounced in the fatigued muscle indicating that aminophylline could be a drug of choice in patients with chronic obstructive lung diseases, where it could guard against development of respiratory muscle fatigue which may contribute to dyspnea and respiratory failure