Microbiological assay of colistin sulfate antibiotic in pharmaceutical formulations

A simple, sensitive and specific agar cup diffusion bioassay for the antibacterial Colistin sulfate was developed. Using a strain of Escherichia coli ATCC 8739 as the test organism, Colistin sulfate at concentrations ranging from 100 to 1600 micro g/ml could be measured in pharmaceuticals. A prospective validation of the method showed that the method was linear [r[2]= 0.999], precise [RSD< 2.8%] and accurate [percent recovery ranges between 98-102%]. The method shows that results confirm its precision, not differing significantly from the other method described in the literature. We conclude that microbiological assay is satisfactory using Escherichia coli ATCC 8739 for quantitation of in-vitro antibacterial activity of Colistin sulfate

Detection and identification of staphylococcus aureus enterotoxins in some milk products and their handlers

Background: Staphylococus aureus may contain one or more genes that encode staphylococcal enterotoxins [SE] that cause food poisoning. The previously known toxins were the five major classical types; however, with the extensive analysis of the S. aureus genome, new genes encoding enterotoxin-like superantigens have been identified. Milk and dairy products are frequently contaminated with enterotoxigenic S. aureus, which is often involved in staphylococcal food poisoning; these contaminations are either from animal or human sources

Aim of the work: To detect the presence and prevalence of coagulase positive S. aureus in milk, kariesh cheese and ice-cream samples and in nasal swabs and stool samples from milk handlers, and to detect types of S. aureus enterotoxins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis [SDS-PAGE], and to detect the type of enterotoxins genes by PCR

Material and Methods: 250 samples of Milk, ice-cream, kariesh cheese, nasal swabs and stool samples from milk handlers were examined for the presence of Coagulase positive Staph aureus, using Mannitol salt agar, Baird-Parker agar, tube coagulase test, and latex agglutination test for protein A and capsular polysaccharides. Confirmed S. aureus isolates were examined for the production of SEs using sodium dodecyl sulphate-polyacrylamide gel electrophoresis [SDS-PAGE], and the type of SE genes by polymerase chain reaction [PCR]

Results: Coagulase positive S. aureus isolates were detected in 82% of Staph colonized raw milk, 80% of Staph colonized ice-cream and 82.3% of Staph colonized kariesh cheese samples and 86.6% of Staph colonized nasal swabs, and 60% of Staph colonized stool samples; with 70.9% of total samples staph colonization exceeds the Egyptian standards. Collectively, 44.3% of coagulase positive S. aureus isolates were enterotoxigenic and the highest percentages were detected in raw milk taken directly from animals [75%] and kariesh cheese from street distributors [66.6%]. In all samples, the major classical enterotoxin genotype was SEA which was detected in 44.4% of toxigenic isolates. SEC was detected in 22.2% of isolates and SED in 18.5% of isolates. SEB could not be detected. For the newly described genes, SEG was detected in 7.4% of isolates and SEH in 7.4% of isolates

Conclusions: Raw milk and some dairy products in the markets in Assuit Governorate rural areas-Egypt, are contaminated with enterotoxigenic S. aureus. The most common type in both milk and dairy products as well as in nasal swabs and stool samples was SEA which is known to be less common among strains from animal origin than from human. Nasal and fecal carriage in human milk handlers is considered a primary source of contamination of milk and dairy products

Selective IgA deficiency among children with recurrent infection

Aim of the study: to evaluate the incidence of IgA deficiency among children suffering from recurrent infections that were suspected to have a primary immunodeficiency disorder from the Assiut Pediatrics University Hospitals, in a hope that this would be the first step to establish a wider epidemiological study including the whole suspected primary immunodeficiency [PID] patients in the country

Methods: This study included 80 cases suffering from recurrent infections that were suspected to have a PID disorder from the Assiut Pediatrics University Hospitals. And eight apparently normal infants and children, age and sex matched that were enrolled as control in the period from July 2011 to May 2012. All children included in the study were subjected to the initial evaluation of immunocompetence which includes a clinical history and physical examination, and investigated for complete blood picture, ESR, Plain chest X-ray when indicated, Screening for serum level of IgA by Enzyme Linked Immuno-sorbent Assay [ELISA] and Radial Immuno-Diffusion [RID]. Patients with proved IgA deficiency were assayed for IgG and IgM

Results: Out of the 80 patients studied, 4 [5%] patients [group I] had laboratory evidence of IgA deficiency; three of them had IgG and IgM within normal level so they were diagnosed as selective IgA deficiency and one [25%] patient had low IgG and normal IgM level so that was diagnosed as common variable immunodeficiency disorder [CVID]. The mean values of age at onset of symptoms, age at diagnosis and the diagnosis lag were 10, 37.25 and 27.25 respectively

The diagnosis delay ranges from 2 months to 7 years. Parents' consanguinity was evident in our study, since 3/4 [75%] patients of group I were the products of consanguineous parents while the ratio was lesser in group II18/76 [23.7%] patients. The current study showed that patients with severe infections had significantly lower IgA levels, younger age and had earlier onset of recurrent infections than patients with less severe infections. There was no specific sex predilection in patients with IgA deficiency. There were no significant differences in both the total leucocytic, absolute lymphocyte and neutrophil counts or hemoglobin level in patients with selective IgA deficiency when compared to patients with normal IgA levels. The duration of infections was significantly longer in patients with selective IgA deficiency [mean: 11.5 days] when compared to patients with normal IgA levels [mean: 9.18 days]

Relationship between coxsackie B-4 virus infection and type-1 diabetes in children

Type-1 diabetes mellitus is believed to result from the selective autoimmune destruction of pancreatic islet cells, occurring in genetically predisposed subjects, and possibly triggered or accelerated by environmental agents. One of the environmental risk factors identified by several independent studies is represented by enteroviral infection. The virus was isolated from infected islets of patients, sequenced, and identified as a Coxsackie B4, In addition, isolated virus was able to in vitro infect cells from non-diabetic multi-organ donors, causing cell dysfunction characterized by impaired glucose stimulated insulin release. We tried to detect the incidence of virally caused type one Diabetes Mellitus with the Coxsackie B4 among cases in Pediatric Department in Assiut University Hospitals. Fifty three cases with recent onset diabetes and 25 matched controls were recruited for the study. We used different techniques to find our targets, including Tissue Culture and PCR. The search yields positively for CVB4 in 54.7% of cases and 4% of controls. Coxsackie B4 may be one of the most common causes of type I Diabetes Mellitus and should be considered in treating of these patients

Studies on Shiga-toxins [STX1 and STX2] produced by escherichia coli isolated from diarrheal cases in Assiut university children hospital

Shiga toxin producing E. coli. [STEC] are emerging pathogens capable of producing sporadic and epidemic diarrhea that may be complicated by haemorrhagic colitis and life threatening hemolytic uremic syndrome [HUS]. The goals of the present study is testing stool specimens for Shiga toxins [Stx1 and Stx2] and detection of the prevalence of the Shiga toxin producing E. coli [STEC] among the diarrheal cases collected in Assiut University Children Hospital. For these purposes, stool specimens from 150 inpatients and 150 outpatients with diarrhea, watery diarrhea or bloody diarrhea and from 40 infants and children with no gastrointestinal illness as control were collected from Assiut University Children Hospital. Escherichia coli was detected in 30 [20%], 64 [42.67%] and 12 [30%] of the inpatients, outpatients and control group respectively. The detection of the Shiga toxins [verotoxins] was done by phenotypic method [Vero cell cytotoxicity assay], immunological method [EIA using RIDA screen kit] and finally by genotypic method [multiplex PCR]. During our study, multiplex PCR [as gold standerd] showed that 4 [13.33%] and 17[26.56%] of E. coli isolated from the inpatients and outpatients respectively were STEC. Non of the E. coli isolateded from control group were STEC. Seven [33.3%] of STEC isolates carried Shiga toxin 1 [Stx1] genes and 14 [66.7%] of STEC isolates carried Shiga toxin 2 [Stx2] genes

Multiple level intercostals block versus paravertebral nerve block for management of chronic thoracic pain

Chronic thoracic pain is one of the most encountered problem in patients having malignancy.ln addition to drug therapy there are different interventional techniques for management of this pain. The aim of this work is to evaluate the analgesic efficacy of the paravertebral block versus multiple level intercostals block in the management of chronic malignant thoracic pain and to study the radiological spread of the dye in both techniques putting into consideration the effect on the pulmonary function and haemodynamics, and the sympathetic block. This study was conducted in the pain clinic in the National Cancer Institute on 60 patients of both sexes having chronic thoracic pain of malignant origin on oral opioid therapy. The patients were randomly allocated into 2 groups each group 30 patients.Group A received paravertebral single injection, whereas group B received three level intercostals blocK.Each group is further subdivided into 2 subgroups to receive either 8 or 12 ml of bupivacaine 0.25%. The study showed that in group a spread of the dye was to 2-4 intercostal spaces with both epidural and contralateral spread whereas in group B the dye was restricted to each intercostal space. Sympathetic blockade in group A lasted 16-17 hours and was absent in group B. Systolic, diastolic blood pressure and heart rate were the same in both groups. Pulmonary function was moderetly higher in group A. There was low VAS score in group A. Paravertebral is better than intercostals block in response to chronic thoracic pain as the injected drug spread for more than one intercostal space with both epidural and contralateral spread and with sympathetic blockade. Paravertebral technique cannot be used as a sole technique for management of this pain

surgical benefits of cervical block anesthesia for carotid endartrectomy

This randomized study was performed to evaluate the benefits of cervical block anesthesia on the surgical technique and outcome of carotid endarterectomy [CEA] aiming to develop a safe and reliable anesthetic and surgical approach for patients undergoing CEA. From May 1997 to May 2001, 36 patients indicated for CEA due to the presence of internal carotid stenosis were randomized into two groups: In group I [18 patients], CEA was done under cervical block anesthesia; while in group II [18 patients], CEA was done under general anesthesia. Hemodynamic measurements and stump pressure were estimated for all cases. The need for shunt, operative time, early postoperative complications and hospital stay was evaluated in both groups. In group I, there was more stable systolic and diastolic blood pressure during the operation. 89% of group I patients tolerated carotid cross clamping without the need for carotid shunt and this method was more accurate than the measurement of stump pressure. Shunt was used in 38.8% of group II patients and this resulted in an intimal dissection in two cases. The operative time was significantly less in group I than group II and discharge from the ICU was earlier

Mycological and therapeutic studies on tinea capitis

A total of 321 cases of tinea capitis representing scaly [60%], favus [29%], kerion [10%] and black dot [1.4%] clinical types were studied. The microscopic and cultural examinations were positive in 63% and 84.5%, respectively. 267 strains of dermatophytes representing Trichophyton violacium [106 strains], Trichophyton schoenleinii [77], Trichophyton mentagrophytes [5], Microsporum canis [68] and Microsporum audounii [11] were recovered. The hair brush and soil- bait techniques seem to be much better in the isolation of T. Schoenleinii and M. audounii. M. canis was isolated by direct method more frequently than the other methods, M. Audounii was only isolated by using the hair brush and soil-bait procedures. The growth of all tested strains was inhibited at different concentrations of dehydroemetine. T. mentagrophytes and T. schoenleinii were more sensitive, while M. canis was least sensitive. DHE was prepared in 0.8% concentration to form an ointment which was used for treatment of 15 selected patients. After three-six weeks, all lesions were cured completely. Clearance of infection was detected mycologically and histopathologically

Serology of human candidiasis in Egypt

A total of one hundred and fifty-two strains of different Candida species was obtained from five hundred and ten human aces of candidiasis. The isolates could be identified as 67 Candida albicans, 20 C. Krusei, 18 C. tropicalis, 10 C. utilis, 9 C. pseudotropicalis, 8 C. parapsilosis, 6 C. lipolytic; 5 each of C. guilliermondii and C. stellatoidea and 4 C. tenusi. Experimental infection in mice showed that C. albicans and C. Krusei were most pathogenic than other species. The study of immune response after experimental injection in rabbits with the most pathogenic strains of each Candida species by agglutination, double diffusion, passive hemagglutination and ELISA was carried out. All the techniques were done using the whole cell antigen and extracts with ether, petroleum ether, chloroform and ethanol. Cross antigenic reaction was demonstrated between the six Candida species. Using the agglutination, precipitation and hemagglutination techniques were able to detect specific antibodies in eight out of seventy-five patients serum samples. The number of serologically positive cases was higher [22.6%] by using the ELISA method