Different detection methods of virulent Helicobacter pylori in gastric biopsies

Background: The global allocation and high level of frequency with the consequence of related pathologies make the eradication of Helicobacter pylori is a very useful advance to test and treat. In addition, the determination of the genotype of H. pylori isolates helps us to comprehend the correlation between assumed virulence genes and clinical disease out come

Objectives: To determine the prevalence of H. pylori infections in patientscomplaining of gastric disorders, the best phenotypic method for detection of H. pylori and the antimicrobial susceptibility patterns among the isolated strains. To compare between phenotypic and genotypic detection methods and to evaluate the frequency of vacA, cagA and iceA genotypes with their clinical outcomes

Methodology:This study was carried out by collecting gastric biopsy endoscopic specimens from 92 participants admitted to Internal Medicine Endoscopy Unit, Faculty of medicine, Menoufia University. Direct detection of H. pylori in gastric biopsy specimens was done by polymerase chain reaction[ureA gene], microaeroplillic culturing, histological examination and Campylobacter like organism [CLO] rapid urease test. Antimicrobial susceptibility patterns among the isolated strains were determined by disc diffusion method. Some virulence genes [cytotoxin-associated gene [cagA], vaculatingcytotoxin [vacA] alleles; vacAs1, vacAs2 and vacAm, also induced by contact epithelium [iceA]] were determined using multiplex PCR

Results:H.pylori genome [UreA] detection by conventional PCR was used as the confirmatory diagnostic tool with 70 PCR positive isolates from 92 participants totally by 76.1%. Histo-pathological examination by both H and E and Giemsa stain detected H. pylori in 68 cases [73.9%]. CLO rapid urease test detected H.pylori urease activity in 64 cases [69.6%]. Microaerophilic culturing detected H. pylori growth in only 32 cases [34.8%].About 100%, 68.8%, 81.3%, 68.8% and 12.5% of isolates were resistant to metronidazole, amoxicillin, tetracycline, clarithromycin and ciprofloxacin respectively.CagA was identified in 58 isolates [82.9%], iceA in 38 [54.3%], vacAs1 in 22 [31.4%], vacAs2 in 10 [14.3%], vacAm in 32 [45.7%]. CagA and cagA+vacAs1m1+IceA were the most prevalent genotypes

Conclusion: Egypt is among the countries that reported high prevalence rate of H.pyloriinfections mainly with antibiotic resistant virulent strains

Tumor necrosis factor-related apoptosis inducing ligand in systemic lupus erythematosus patients; potential role in disease activity, neutropenia and renal impairment

Background: The role of tumor necrosis factor-related apoptosis-inducing ligand [TRAIL] in immunopathogenesis of systemic lupus erythematosus [SLE] was previously documented. Neutropenia and nephritis are common features in SLE patients and have been hypothesized to be due to accelerated apoptosis induced by binding of death receptor ligands like TRAIL to their cognate receptors on sensitive cells

Objectives: The aim of this study was to determine the serum concentration of soluble TRAIL [sTRAIL], the expression level of TRAIL mRNA in the peripheral blood mononuclear cells [PBMC] and expression level of TRAIL receptor-1 [TRAIL-R1, death receptor-4/DR-4/CD261] on polymorphonuclear leucocytes and to clarify their relation with disease activity, neutropenia and renal impairment in SLE patients

Methodology: The study enrolled 25 patients with active SLE, 25 patients with mild or no disease activity, 25 patients with Rheumatoid arthritis and 15 age and gender-matched healthy volunteers as a control group. Serum level of circulating TRAIL was measured by ELISA, the expression level of TRAIL mRNA on PBMC was determined by Quantitative Real-Time reverse transcription-polymerase chain reaction and flow cytometry was applied to evaluate the expression level of TRAIL R1 on polymorphonuclear leucocytes among the study population

Results: Serum level of sTRAIL was significantly [P<0.001] higher in patients with active SLE than those with no activity, Rheumatoid arthritis and healthy controls. Up-regulation of TRAIL mRNA expression in the PBMN cells and TRAIL R1 expression on neutrophils was detected in active lupus patients with a statistically significant difference [P<0.001] compared to other participants. A statistically significant correlation was detected between sTRAIL, TRAIL mRNA and TRAIL R1 expression and SLE activity, neutropenia and nephritis [P<0.001].The sensitivity, specificity, PPV, NPV and accuracy of TRAIL mRNA were 80%, 60%, 66.7%, 75% and 70% respectively

Conclusion: Concentration of circulating TRAIL and TRAIL mRNA levels could be potential markers for SLE activity assessment and predictors of lupus-associated neutropenia and renal affection

Fungal rhinosinusitis of the immunocompetent host: correlation study between computed tomography scan, mycological and histopathology

Background: Fungal rhinosinusitis is a characteristic disease that requires a great deal of interest. Knowing the fungal flora, its prevalence and symptomatic presentation in patients with chronic fungal rhinosinusitis will allow a better understanding of this disease, correct diagnosis, and treatment and developing its prognosis. Clinical presentation can provide to the subcategories of fungal rhinosinusitis however, histopathological and microbiological examinations provide accurate diagnosis and classification

Patients and Methods: This study was conducted on fifty patients of chronic fungal rhinosinusitis who had been referred to otorhinolaryngology surgeon for endoscopic sinus surgery in the last 2 years in Dr. Soliman Fakeeh Hospitals in Jeddah, Kingdom of Saudi Arabia, We selected some immunocompetent chronic rhinosinusitis patients with signs and symptoms of inflammation of nasal and paranasal sinuses. These patients had positive computed tomography and /or histopathological examinations. We evaluated clinical history, otolaryngologic examination with nasal endoscopy, computed tomography scan, mycological and bacterial cultures and histopathological examinations

Results: Fungal rhinosinusitis was the cause of chronic rhinosinusitis in 16.2% of patients with chronic rhinosinusitis submitted to paranasal sinuses endoscopic surgery. Fungal cultures were positive in 60% of specimens with predominance of 63.3% Aspergillus fumigatus, 20% Aspergillus flavus, 3.33% Aspergillus niger and 13.33% Candida albicans. While 40% of patients with rhinosinusitis showed no fungal growth. Bacteriological cultures indicated there is an association of bacterial infection in 16 patients out of 50 as; Staphylococcus aureus [43.75%], Staphylococcus haemolyticus [25%], Pseudomonas aeruginosa [18.75%] and klebsiella pneumonia [12.5%]. In 28% specimens there was no bacterial growth, and in 40% specimens the bacterial examination were not performed. Mixed bacterial and fungal infections were found in 30% as the following: Staphylococcus aureus and Aspergillus fumigatus, Staphylococcus aureus and Aspergillus flavis, Pseudomonas aeruginosa and Aspergillus fumigatus, Klebsiella pneumoniae and Candida albicans, and Staphylococcus haemolyticus and Candida albicans in 33.33%, 22.22%, 22.22%, 11.22% and 11.22% respectively. According to the histopathological findings the detected types were fungal rhinosinusitis, allergic rhinosinusitis, non specific inflammation and mixed reaction in 54%, 22%, 6%, and 18% respectively. All patients presented some type of findings in paranasal sinuses by computed tomography [CT] scan were classified as 60% allergic fungal sinusitis, 34% chronic invasive fungal sinusitis, 4% fungal ball and 2% acute invasive fungal sinusitis. As regards correlation of histopathology, CT and fungal cultures results of the studied 50 patients and according to CT classification of fungal sinusitis, positive histopathological findings were found in 53.33% of cases that were classified as allergic fungal sinusitis, while positive fungal culture were seen in 40%. In chronic invasive fungal sinusitis, histopathological findings were positive in all cases [100%] while positive fungal cultures were seen in 88.23%. In acute invasive fungal sinusitis and fungal ball CT classification, both histopathology and fungal cultures were positive in all cases [100%]

Aim of work: The aim of this study is to analyze and compare the results of clinical endoscopic findings, radiological, mycological, and histological criteria for optimizing the diagnosis of true fungal sinusitis

Conclusion: Early and specific diagnosis of fungal rhinosinusitis is necessary. The traditional methods used in routine practice for the diagnosis of fungal rhinosinusitis may be insensitive and nonspecific. Moreover, the allergic fungal rhinosinusitis represents an immunologic rather than infectious disease. The optimal duration of treatment and the role of patient preferences in clinical decision making also needed to be addressed. The maximum diagnosis will be available by combining traditional culture, histopathology and radiology. In this circumstance, molecular techniques are perhaps best placed to enable rapid and accurate identification

Surveillance of aminoglycosides resistance among proteus mirabilis isolates from different units in Jeddah hospitals, Saudi Arabia

Proteus mirabilis isolates had undergone considerable alteration in susceptibility to aminoglcosides gradual increase to resistance in gentamycin and tobramycin. Fortunately, amikacin have a broad spectrum of activity and is particularly valuable in treating nosocomial infections so this study aimed at obtaining data on susceptibility patterns of Pr. mirabilis isolates responsible for hospital infections in Jeddah, Saudi Arabia to the currently used aminoglcoside agents in their treatment. Results showed that the most predominant organisms isolated from hospital-acquired infection were Gram-negative [60.93%], mostly isolated from urine [45.73%], where, Pr. mirabilis isolates accounted for 162 out of 1383 [11.7%] Enterobacteriaceae. About 89.6%, 84.3% and 82.8% of Pr. mirabilis isolates were sensitive to amikacin, tobramycin and gentamycin respectively by disk diffusion method. The highest level of resistance was found in Pr. mirabilis strains isolated from pus. The resistance of Pr. mirabilis isolates to aminoglycosides was allocated in 6 different patterns, and the most predominant pattern was pattern III [49.5%]. MICs of most Pr. mirabilis isolates when detected by broth dilution method gave higher readings than when detected by agar dilution method and the disk diffusion method gave results similar to that of agar dilution. Only 30 out of 192 [15.6%] of Pr. mirabilis isolates harboured one plasmid on each strain and about 90% of Pr. mirabilis isolates resistant to gentamycin, tobramycin and amikacin were containing plasmid. Sixty percent of plasmid-containing Pr. mirabilis isolates were able to transfer their antibiotic resistance to the standard E.coli at a frequency ranged between 3x10-8 and 1.2x10-7. The resistance to amoxicillin, ampicillin, cephalothin, neomycin, streptomycin, kanamycin, gentamycin, tobramycin and amikacin was transferred in 100% of isolates due to presence of plasmids with molecular weight 180, 170 and 25 Kb. In conclusion, the present study shows that aminoglycosides that are commonly used in treatment of infections caused by Pr. mirabilis isolates are still effective. The MIC by disk diffusion method gave result similar to that obtained by agar dilution method; moreover, it is easy, cheap and rapid. Indiscriminate use of antibiotics has led to selection of resistant strain with R-plasmids which are transferable between enteric bacteria by conjugation and these resistant bacteria disseminate among individuals

Helicobacter pylori in diabetes mellitus: value of different diagnostic methods

Helicobacter pylori is one of the most common chronic bacterial infections in diabetic patients due to alteration of glucose metabolism, abnormal emptying of the stomach and autonomic neuropathy. So, this study was done to determine H. pylori infection in diabetes, to evaluate the different diagnostic methods of its infection and to estimate the efficacy of he current antimicrobial therapy to eradicate H. pylori. The study involved 40 patients with diabetes mellitus [12 with type 1 and 28 with type 2], and 20 non-diabetic patients as control group. Their sera were used for the assay of H. pylori specific lgG and IgA by enzyme immunoassay [EIA]. Biopsy specimens were tested by rapid urease test [RUT], stained by Gram's stain and cultured to isolate H. pylori. Antimicrobial susceptibility testing was performed using Epsilometer test [E-test]. Results showed that there was a significant increase in the incidence of H. pylori in diabetic patients, 77.5% by RUT, 75% by culture, 70% by lgG, 50% by IgA and 20% by direct smear. There was a significant association between age and smoking and the presence of IgG and IgA. The specificity of the direct smear and IgA was 100%, the sensitivity of RUT was 100%, positive predictive value [PPV] of direct smear and IgA was 100%, negative predictive value [NPV] of RUT was 100%. H. pylori was highly sensitive to tetracycline [93.3%] and amoxicillin [73.3%] and relatively resistant to metronidazol [70%]. We concluded that H. pylon infection is more common among diabetic patients. Culture is the method of choice in diagnosis of H. pylori as it allows testing for antimicrobial therapy. EIA for anti-H. pylori lgG could be of value in screening population and in excluding H. pylori negative infections. Anti-microbial sensitivity must be performed for better eradication of H. pylori in diabetic patients

Hand washing practices and efficacy of alternative hand disinfectants in health care [Menofia University Hospital]

A cross sectional interventional study was conducted on a random sample of 120 nurses in medical and surgical wards. Data were collected through a predesigned questionnaire and checklist. The number and types of bacteria on hands of nurses before and after disinfection with alternative agents and after drying were evaluated by the standard microbiological method. The results revealed that the compliance of nurses with hand washing between patients contact was quite poor [1.7%] which referred mainly to the work overload [94.9%] and the lack of resources [39.8%]. The majority of nurses had transient bacterial flora [97.5%]. The frequency of Staphylococcal aureus, E. coli, Klebsiella and Streptococcal faecalis hand infections were 47.5%, 30%, 5% and 4.2%. Betadine and alcohol disinfectants had the highest efficacy [95.5% and 90% in medical ward and 90.9% and 82.9% in surgical ward, respectively]. The efficacy of alternative hand disinfectants generally was improved after hand drying with paper towel. For plain soap and tap water, the efficacy after drying became nearly similar to betadine and alcohol

Urinary tract infection in diabetic patients with special reference to klebsiella species

Diabetes mellitus [DM] is a metabolic disease which is accompanied by increased susceptibility to infection. This study was performed to determine the prevalence of urinary tract infection [UTI] in a group of diabetic patients and to identify the most common causative organisms with special reference to Klebsiella species and to assess antimicrobial sensitivity of the isolated strains of Klebsiella. The study included 618 diabetic patients along with 100 age- and sex- matched non-diabetic individuals as controls. Urine was obtained by clean catch midstream method. Bacterial colony count and culture on blood agar and MacConkey agar were performed. Isolation and identification of the causative organisms were done by the standard microbiological techniques. Antibiotic sensitivity was done for the isolated Klebsiella strains using agar dilution technique. Results showed an increase in prevalence of significant bacteriuria in diabetic [30.91%] than in non-diabetic individuals [13%] with a higher frequency of infection in females and in older age group. There was no significant difference between both types of diabetes. However, patients with longer duration of disease [25.69%] were significantly [P < 0.001] more liable to have significant bacteriuria than those with shorter duration of the disease [38.28%]. Most diabetic patients with UTI were asymptomatic [69.63%] and the most common organisms found were E. coli [32.22%] and Klebsiella [23.33%] with prevailing K. pneumoniae followed by K. oxytoca subspecies. Most Klebsiella strains were resistant to the commonly used antimicrobial drugs. However, they were mostly susceptible to aztreonam, amikacin, nalidixic acid and third generation cephalosporins. In conclusion, UTI is common and often asymptomatic in diabetic patients and is mostly caused by a highly resistant species indicating the importance of doing culture and sensitivity to select the most suitable antimicrobial drug for treatment of these patients