Evaluation of different methods for the detection of KPC b-lactamase in Klebsiella pneumoniae and escherichia coli

The worldwide increase in the occurrence and dissemination of Klebsiella pneumoniae carbapenemase [KPC] among Gram-negative pathogens makes critical the early detection of these enzymes. The aim of this study is the evaluation of the most appropriate method for the detection and identification of KPC producing Klebsiella pneumonia and Escherichia coli. Modified Hodge test [MHT], Boronic acid lest [BAT] and polymerase chain reaction [PCR] were evaluated for the detection of KPC-possessing isolates. A total of 60 urine samples and 60 sputum samples were collected from 79 patients in Chest Intensive Care Unit and Trauma intensive Care Unit in Assiut University Hospital. Sixty four isolates [26 Klebsiella pneumonia and 38 E. coil] were identified by conventional methods. The antibiotic susceptibility testing of isolates showed that 19% were resistant to meropenem, 17% were resistant to imipenem and 95% were resistant to ertapenem. Thirty seven isolates were MHT positive while twenty three only of them were Boronic acid and PCR positive. The results indicate that the MHT lack sensitivity while Boronic acid test is an excellent inhibitor of KPCs. PCR is the standard method for the detection of the KPCs but it's highly expensive

Prevalence of efflux mediated ciprofloxacin and levofloxacin resistance in pseudomonas aeruginosa urinary isolates

Pseudoinonas aeruginosa [P. aeruginosa] represents a phenomenon of antibiotic resistance, and demonstrates practically all known mechanisms of bacterial resistance. Active efflux is an important mechanism of resistance in P. aeruginosa. It contributes to the development of multiple resistances to all strategic antipseudonional antibiotics. More than five hundred urine samples were collected from patients in Assiut University Hospital. Fifty P. aeruginosa isolates were identified by conventional methods. The antibiotic susceptihity testing of isolates showed that 68% of isolates were resistant to ciprofloxacin and 62% were resistant to levofloxacin. A comparison between the effect of three efflux pump inhibitors [Reserpine, Pantoprazole and Carbonyl cyanide m-chlorophenylhydrazone [CCCP]] on the activity of ciprofloxacin and levofloxacin was done by measuring ability of these agents to potentiate effect of ciprofloxacin and levofloxacin against resistant P. aeruginosa isolates. Outer membrane profile of P. aeruginosa isolates was also done using sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE]. Reserpine was able to potentiate effect of ciprofloxacin in 50% of isolates, and in 5.5% for levofloxacin. Pantoprazole results were 33.3% for ciprofloxacin, 16 7% for levofloxacin. Finally CCCP potentiate. Regarding the SDS-PAGE of P. aeruginosa isolates, all isolates produced proteins with apparent molecular masses in the range of 50-54kDa.Reserpine-ciprofloxacin proved to be the best combination against multidrug resistant P. aeruginosa. Over production of 50-54 KDa outer membrane proteins is responsible for emergence of P. aeruginosa strains highly resistant to fluoroquinolones in clinical settings

Biofilm formation by e-coli causing catheter associated urinary tract infection [CAUTI] in Assiut university hospital

Urinary tract infections [UTIs] are the most common cause of nosocomial infection and up to 80% of UTIs are associated with the use of urinary catheter. Inappropriate use of this device may lead to an increase incidence of infectious complications. It has been estimated that 65 % of nosocomial infections are biofilm associated urinary tract infections, loading the health care system enormous costs. These biofilm infections are 10 to 1000 times more resistant to the effects of antimicrobial agents. In this study urine samples were collected from 150 patients with CAUTI [group 1] giving one hundred and fifty bacterial isolates and 70 non catheterized UTI patients [group 2] giving fifty bacterial isolates. Out of the two hundred isolates the most common isolated pathogens were: Escherichia coli[E.coli] [50%] in group I, [48%] in group 2, followed by Klebsiella [26.7%] in group J, [28%] in group 2, pseudomonas aeruginosa [8%] in group 1, [12%] in group 2 then Staphylococcus aureus [8.6%] in group 1, [4.%] in group 2, Proteus [4.6%] in group 1, [4%] in group 2, and lastly, Candida albicans [2%] in group 1,[4. %] in group 2. The E.coli isolates were evaluated for biofilm formation using congo red agar [CRA] and microtitre plate methods. Out of 99 E.coli isolates; 27 were non biofilm forming in group 1, 19 isolates in group 2 while 48 isolates were biofilm forming in group 7, only 5 isolates in group 2 . Using microtitre plate method; out of 48 biofilm forming isolates in group 1; 8 isolates [16.6%] were weak biofilm forming, 10 [20.8%] were moderate biofilm and 30 isolates [62.5%] were strong biofilm forming while, 3 isolates were weak biofilm forming in group2. The two methods used to detect biofilm formation [CRA test and spectrophotometer], both are valid tests. CRA is simpler but spectrophotometer differentiates between weak and strong biofilm producers

Short term effect 1% topical phenytoin suspension on the pain and wound dimensions of donor site after free gingival grafts

Ulcer and pain have been the most complications arised after periodontal surgery. The purpose of this study was to evaluate the healing and analgesic effect of Phenytoin in donor site of free gingival grafts. This study is a randomized clinical trial with 10 patients received free gingival graft and divided in 2 groups. The wound size measured with digimatic caliper. Donor sites rubbed in case group with Phenytoin and with placebo in control group. The wound size measured in 14th and 28[th] days and evaluated in comparision with baseline. Also, the pain level measured via VAS. Statistical analysis was performed using T pair test and wilcoxaon signed ranks. Statistical analysis showed that case group had better healing due to standard diviation in both appointments [a=0.1, p=0.078, and a=0.05, p=0.049, respectively]. Pain Score was lower in patients treated with phenytoin [p=0.18]. Phenytoin is effective in healing of donor site and reduce pain

Prevalence of sen virus infection in hemodialysis and blood diseased patients in Assiut university hospitals

SENV is a blood- borne, circular ss DNA virus and possessing nine genotypes [A to I]. Among nine genotypes, SENV-D and SENV-H genotypes have the strong link with patients with non [A-E] hepatitis infections .Recently, the identification of SEN virus [SENV] as a possible etiologic agent of parenteral transmission hepatitis let to the study of the prevalence of such agent. This study compared SENV prevalence and its two important genotypes [D and H] which might be pathogenic in high risk subjects including blood diseased patients and hemodialysis patients and low risk subjects as healthy blood donors. This study included 75 multitransfused patients, 60 of them were hemodialysis and the remaining were blood diseased including hemophilics, anemics and leukemics. The study included also 25 healthy blood donors as a control They were received consecutively at Department of Internal Medicine, Assiut University Hospital. The sera were separated and SENV DNA was detected by polymerase chain reaction. The results showed a higher prevalence of SENV infection in patients group than blood donors [46.7% versus 20%].No significant relation was found between SENV infection and age, duration of hemodialysis or liver enzymes. However, there was significant difference between SENV positive and negative patients as regards gender and number of blood transfusions. SENV is commonly present in blood diseased and hemodialysis patients attended to Assiut University Hospitals as well as in blood donors at variable rates. SENV infection has been found in only 20% of blood donors but in 46.7% of patients. The results also indicated that other possible routes of SENV infection other than blood transfusion may be included. Its pathogenic role in causing hepatitis is not documented, so far it can be considered as simple guest till further studies have been done

Comparison of adsorption process by GAC with novel formulation of coagulation-flocculation for color removal of textile wastewater

This study evaluates the effectiveness of adsorption process by Granular Activated Carbon [GAC] compared with a novel formulation of coagulation - flocculation process for dye removal from textile wastewater. In this regard, acidic, reactive, disperse and direct red dye are used to prepare the synthetic dye. Dominant wave length for each dye is determined by spectrophotometeric method. Using GAC as adsorbent, equilibrium time and adsorption isotherm of each dye are determined with aid of spectrophotometric method. The results show that GAC can not remove dispersed red dye. Acidic red, direct red and reactive red of 5 mg/L concentration are removed by GAC up to 90%, 88% and 43% in 30, 60 and 120 min. [equilibrium time] respectively. Dyes of 50 mg/L concentration are removed up to 93%, 30% and 51% in 15, 90 and 150 min. respectively. Adsorption obeys Freundlich isotherm for acidic red, BET isotherm for direct red and Langmuir isotherm for reactive red. This investigation presents a novel formulation of coagulation - flocculation for color removal from textile dye solutions and illustrates its efficiency. Novel formulation of coagulation - flocculation remove direct red, reactive red and disperse red of 5 mg/L concentration respectively up to 93%, 91.3% and 57.1%. Also the mentioned dyes of 50 mg/L are removed respectively up to 90.8%, 91.9% and 70.1%

Endothelial cell dysfunction and inflammatory process in type 1 diabetic patients with-and without micro vascular disease

Endothelial cell dysfunction results in altered production of cell adhesion molecules [CAMs] that may be involved in the pathogenesis of diabetic microvascular disease. Increased circulating cytokines may also be involved in this process. The aim of the present study was to evaluate levels of some CAMs and cytokines in children and adolescents with type 1 diabetes. It was also aimed to assess these parameters in relation to microvascular complications and certain risk factors. The study included 45 cases with type 1 diabetes aged 8-22 years of whom, 30 cases had evidence of microangiopathy [retinopathy or nephropathy] and 15 cases had not. Fifteen apparently healthy matchable subjects were included as controls. Cases were subjected to full history taking and physical examination. Direct ophthalmoscopy and fluorescein angiography were used to diagnose retinopathy, while nephropathy was diagnosed by detection of microalbuminuria. Level of glycated hemoglobin [HbA[1c]] and serum levels of sVCAM-1 and sE-selectin as well as IL-6 and TNF alpha were assessed for all patients and controls. The results showed that diabetic patients as a whole had significantly higher serum levels of sVCAM-lt sE-selectin, IL-6 and TNF alpha than controls. Post pubertal age, long duration of illness, obesity and high HbA[1c] level were significant risk factors for higher levels of CAMs. Significant positive correlations were found between levels of HbA[1c] and each of serum levels of sE-selectin and IL-6. Also significant positive correlations were found between each of serum levels of sVCAM-1 and sE-selectin, and IL-6 and TNF alpha. Patients with evidence of microangiopathy had significantly higher level of sVCAM-1 than cases without, and the latter group had significantly higher level of sE-selectin than controls. It is concluded that young patients with type 1 diabetes had significant markers of endothelial cell dysfunction particularly in those with microvascular disease. Screening of diabetic patients with E-selectin may help early diagnosis of endothelial dysfunction. Strict glycemic control and new therapeutic targets are mandatory to improve diabetic outcome in such cases

Study of pancreatic islet cell function in cirrhotic patients due to hepatitis C virus

The liver is of key importance in the proper functioning of most of endocrine system. It is a major organ for metabolic degradation of many hormones. In chronic liver disease, there is a change in the concentrations of main pancreatic hormones. In this study, we aimed to study pancreatic islet cell hormones in non ascitic non diabetic cirrhotic HCV patients by measuring these hormones basally and after intravenous glucose load with different concentrations to evaluate pattern of endocrine pancreatic hormonal response at increasing glucose concentration. This case-control study included 25 non ascitic non diabetic patients with cirrhosis due to HCV infection; age ranged from 28 to 60 years. They were selected from outpatient clinic of National Hepatology and Tropical Medicine Research Institute [NHTMRI] during June 2004 to January 2005. Fifteen age and sex matched apparently healthy control subjects from same area were also included in study. Assessment of pancreatic islet cell function was done by assaying insulin, C-peptide, somatostatin and glucagon hormones. All hormones were determined by radioimmunoassay [RIA] for both patients and controls. Basal insulin, basal somatostatin and basal glucagons levels were significantly higher than that of controls [p<0.05]. Basal C-peptide was significantly lower than that of controls [p<0.05]. After intravenous glucose load with different concentrations 5%, 10% and 25%, insulin level showed a significant increase than controls [p<0.05], while other hormones showed no significant difference as compared to controls [p>0.05]. Pancreatic islet cell functional defects with liver cirrhosis due to HCV infection

Polymorphisms of TAP1/LMP7 loci in Egyptian patients with vitiligo

Vitiligo is a very common, often heritable acquired disorder characterized by well-circumscribed milky white cutaneous macules devoid of identifiable melanocytes. Vitiligo appears to be more commonly observed in parts of the body exposed to the sun and in darker skin types and may develop at any age. Our study contained 47 unrelated Egyptian young-aged vitiligo patients [age range 2-18 y] and 14 healthy volunteers of matched age range. All patients experienced active vitiligo lesions with no signs of other autoimmune disorders. To genotype the TAP1 [C>T intron 7] and LMP7 [G>T intron 6], we amplified the genomic DMA using polymerase chain reaction [PCR] using genomic DNA followed by enzymatic digestion. Our results showed no significant difference between TAP1 C>T [intron 7] and LMP7 G>T [intron 6] alleles and healthy controls [p= 0.3 or 0.5, respectively]. Even so, the odd ratios [ORs] for the genotypes of the TAP1 [C>T] were 1.78 [0.4 to 7.0], 0.8 [0.5 to 1.2], and 1.19 [0.2 to 4.9] for the TAP1-CC, CT, TT-genotypes, respectively. The ORs of LMP7 [intron G>T] genotypes were 1.4 [0.3 to 6.0], 0.8 [0.5 to 1.3], 1.19 [0.3 to 3.6] for GG, GT, and TT, respectively. However, a major contribution of both TAP1 and LMP7 polymorphisms to vitiligo susceptibility cannot be excluded. Further studies of other alleles within the TAP and LMP gene regions in Egyptian patients is recommended to demonstrate a possible role for MHC class I antigen processing and/or presentation pathway in the antimelanocyte autoimmune response in vitiligo pathogenesis

clinical picture of hepatitis C virus as a concomitant infection with fascioliasis

Hepatitis C and human fascioliasis are two of the most important public health problems locally and internationally. Each one has its own complications regarding spreading to man, clinical picture, laboratory and serologic diagnosis, treatment and prevention. Concomitant human infection with both magnified the complications. Clinically, both diseases [HCV and fascioliasis] have their own broad signs and symptoms. In concomitant infection, the clinical pictures of both showed some variations. The most common feature in fascioliasis and HCV patients was easy fatigability in 78.6% and the lowest was 7.18% for each of tympanic abdomen, tender colon, tender right upper quadrant, itching, arthritis, epi-gastric pain, and right quadrant pain. The eosinophilia% ranged from 5-24 and the haemogiobin ranged from 7-11.1 gm/dl. In patients with fascioliasis alone, the most come feature was pallor in 96.15% and the lowest was 3.85% for each of splenomegaly, ascites and itching. Eosinophilia% ranged from 1 to 22 and haemoglobin ranged from 6-12 gm/dl. In general, the double infection with both HCV and fascioliasis magnified the laboratory and clinical pictures of such patients

liver profile in patients with hepatitis C virus and/or fascioliasis

In this study, twenty HCV/PCR-RNA positive patients with neither infection nor infestation were 15 males and five females with ages ranging between 17-78 years old. Liver function tests: S. albumin was decreased in 15 patients [75%], total protein decreased in 3 patients [15%], total bilirubin increased in 7 patients [35%], AST/ALT increased in 3 patients [15%]. Globulin value increased in 15 patients [75%]. A/G ratio decreased in 12 [60%]. Ten normal individuals [five males and five females], gave normal findings. However, a 19 years-old female had non significant elevation [0.26mg/dl] in direct bilirubin. Liver function tests: one patient had increased AST/ALT, Globulin value decreased [-0.2] in another one [10%] and A/G ratio increased [+0.3 to+0.6] in three [30%] individuals. Twenty randomly selected patients [15 males and five females] HCV/3 rd generation ELISA showed ALT elevation in 17 [85%], AST in all [100%], S. albumin decreased in 9 [45%], and increased in 1 [5%]. Total protein decreased in 3 [15%], total bilirubin increased in 7 [35%] and direct bilirubin in 4 [20%], AST/ALT value increased in 3 patients [15%], Globulin value increased in 15 patients [75%]. A/G ratio decreased in 12 [60%]. Three patients had schistosomiasis, one 30 years old male had increased AST/ALT, normal globulin, increased A/G and positive HBs-Ag. The second one was a 33 years old male had normal AST/ALT, normal globulin, increased A/G and positive HBs-Ag. The third patient was a 19 years old female with normal AST/ALT and normal globulin, increased A/G and positive HBs-Ag. Two patients had fascioliasis, one was a 20 years old male with increased AST/ALT, globulin normal, A/G normal and negative HBs-Ag. The second one was a 26 years old female with normal AST/ALT and normal globulin, increased A/G and positive HBs-Ag. The other four positive HBs-Ag patients were parasite-free. Also, the other HCV/ELISA positive were negative for HBs-Ag. HCV/ELISA may have cross-reacted with HBs-Ag and/or with elevated ALT and gave false positive HCV. The 3rd generation ELISA in detection of HCV was not as sensitive as PCR/RNA. Out of 41 fascioliasis patients [26 males and 15 females], 14 were positive HCV/ELISA [34.1%], but only six were positive HCV/PCR [14.6%]. ALT increased in 18 fascioliasis patients [43.9%], AST in 23 patients [56.1%] but, albumin decreased in 7 patients [17.1%], total protein decreased in 5 [12.2%], but total bilirubin increased in 14 [34.1%] and direct bilirubin increased in 2 [4.9%]. Liver function tests of 14 fascioliasis and ELISA positive HCV showed AST/ALT increased in 6 [42.9%], globulin increased in 3 [21.4%] and decreased in 6 [42.8%]. A/G decreased in 4 [28.6%] and increased in 8 [57.2%]. Liver function tests of pure 27 fascioliasis patients showed that AST/ALT increased in 8 [29.6%], globulin increased in one patient [3.7%] but decreased in 10 [37.0%] and A/G ratio increased in 13 [48.1%]

HCV/PCR positivity in bile and doudenal aspiration of Fascioliasis and/or HCV patients

In this study, three bile aspirates taken from 10 fascioliasis patients [30.0%] showed HCV positivity by PCR/RNA. Also, four duodenal aspirates [66.7%] taken from six HCV/PCR RNA positive patients and three duodenal aspirates [20%] taken from 15 pure fascioliasis patients showed HCV positivity by PCR/RNA This is the first time to demonstrate HCV/PCR-RNA in the bile and duodenal aspirates of fascioliasis patients and in the duodenal aspirates of HCV patients. So, PCR can be used for the detection of HCV in the bile and/or duodenal aspirates of HCV suspected patient. On the hand, this outcome results may incriminate HCV infection as concomitant with fascioliasis or incriminate fascioliasis as paving the way to HCV.

liver function profile in PCR-RNA Egyptian HCV-patients and normal controls

No doubt. Hepatitis C virus [HCV] is a real health problem worldwide. The liver function tests [S.ALT, S.AST, Albumin, Total Protein, Total Bilirubin and Direct Bilirubin] were evaluated in 20 PCR-RNA positive HCV-patients and 10 cross matched apparently healthy population. All the HCV-patients and controls were free from liver helminthes. The results showed that in the HCV-patients, there was elevation in the level of S.ALT [17/20 or 85%], S.AST [20/20 or 100%], Total Bilirubin [7/20 or 35%], and [4/20 or 20%]. Besides, there was neither a correlation between sexes nor the degrees of viraemia and the elevation of these four parameters. However, serum levels of Albumin, and Total Protein were within the normal range. On the other hand, in the controls the levels of the six tests were within the normal range. Nevertheless, only one control subject who had positive HBs-Ag, showed elevated Total Bilirulin and Direct Bilirubin. Consequently, these tests are indicative as useful and dependable markers in the noninvasive diagnosis of the hepatitis C virus [HCV]

Plasma fibronectin and serum complement c3 levels in chronic active hepatitis following virus B versus virus C infection

Plasma fibronectin and serum complement C3 levels were estimated in 30 patients with chronic active hepatitis [CAH] post virus B and another 30 patients with CAH post virus C as well as 20 normal healthy subjects. Fibronectin level was significantly increased in CAH when compared to normal controls. Moreover, fibronectin level was significantly increased in CAH following virus C when compared to CAH following virus B hepatitis. Concerning complement C3 there was no significant changes in the different groups studied. It is concluded that CHA following virus C may lead to vigorous inflammatory damage than CAH following virus B infection

case of oriental diagnosis as pseudolymphoma cutis

A parasitologically positive case of oriental sore was diagnosed pathologically as pseudolymphoma cutis. The case was successfully treated with Astiban