Phenotypic and genotypic detection of some antimicrobial resistance mechanisms among multidrug-resistant acinetobacter baumannii isolated from immunocompromised patients in Egypt

Acinetobacter spp. are Gram-negative bacteria that have become one of the most difficult pathogens to treat. The species A. baumannii, largely unknown 30 years ago, has risen to prominence particularly because of its ability to cause infections in immunocompromised patients. It is now a predominant pathogen in many hospitals as it has acquired resistance genes to virtually all antibiotics capable of treating Gram-negative bacteria, including the fluoroquinolones and the cephalosporins. The aims of this study were to [i] evaluate the antimicrobial susceptibility of Acinetobacter baumannii isolated from immunocompromised host [ii] phenotypic assessmentof the prevalent mechanism of resistance among multi drug resistant Acinetobacter spp. [iii] evaluate the prevalence of metallo beta-lactamases [MBLs] phenotypically and genotypically among multidrug resistant A. baumannii. Seventy-two nonreplicated A. baumannii isolates were recovered from 631 clinical specimens referred for bacteriological cultures from immunecompromised patients of all age groups and both sexes admitted in El-Demerdash Hospital and National Cancer Institute [NCI] Cairo-Egypt. Isolates were identified conventionally using standard biochemical tests and also using Microscan [Dade Behring, West Sacramento, USA]. Both manual [following the CLSI protocol] and automated methods [using Microscan system] were used to detect antimicrobial susceptibility pattern of Acinetobacter isolates. Different potential resistance mechanisms were investigated in 45 carbapenem resistant A. baumannii phenotypically. Genotypic detection of MBLs was carried out using PCR. Acinetobacter baumannii represents the predominant Acinetobacter isolates [83.3%]. The mean age group of patient with Acinetobacter infection was more than 55 years old [36.1%]. A. baumannii exhibited high resistance rate to the majority of commercially available drugs including imipenem [66.6%], meropenem [73.3%] andcefazolin and cephalothin [100%]. Isolates show moderate susceptibility to tetracycline [40% of the isolates were susceptible] and gentamicin [33.3%]. MDR A. baumannii represent 75% of the Acinetobacter isolates [45/72].Using phenotypic tests; none of the carbapenem-resistant A. baumannii were carbapenemase producers and44.4%wereAmpC beta-lactamase positive. MBLs were detected in 55.6% using phenotypic tests and in 44.4% of isolates using PCR. Moreover Efflux pump was detected in 77.8% of isolates. Multidrug resistant A. baumanniiis a problematic organism in immunosuppressed patients since it became resistant to the majority of commercially available antimicrobials with different resistance mechanisms. Metallo-beta-lactamase production is an important mechanism of carbapenem resistance and AmpC - beta-lactamase could be a contributory factor for meropenem resistance among MDRAB isolates. This underlies the importance of their accurate identification and reporting to prevent the emergence of absolute resistance to the useful drugs against Acinetobacter spp. in Egypt

Ewing's sarcoma family tumors in the western region of Saudi Arabia. A pathological experience from 2 tertiary medical centers

To review the pattern of Ewing's sarcoma/primitive neuroectodermal tumor [ES/PNET] in 2 medical centers in the western region of Saudi Arabia. We retrospectively analyzed the pathological data of patients diagnosed with ES/PNET in 2 tertiary medical centers in the western region of Saudi Arabia [King Abdulaziz University Hospital, [March 1995 to November 2011], and King Faisal Specialized Hospital [April 2003 to 12 December 2010]]. Age, gender, and site of tumors were analyzed. Sixty-nine cases were diagnosed as ES/PNET. The age range was 3-62 years [mean 22 years]. Male cases were more than the female. Approximately 28.9% of cases presented within the skeleton, and 71.1% cases were presented as a soft tissue disease. Bone affection was higher in the iliac bone. Long bones were affected at a lower frequency. Soft tissue affection showed a higher incidence in the head and neck region followed by the lower limb. The current study represents a review of a large number of Ewing's sarcoma family of tumors in western Saudi Arabia. Cases showed clinicopathological features comparable to those reported from other locations worldwide apart from relatively higher soft tissue affection than skeletal affection and a higher incidence of head and neck involvement by soft tissue ES/PNET. Further, multicenter studies [epidemiological and genetic] are recommended to obtain profiling of the disease and effect on outcome and therapy

Sleep-wake cycle disturbances in protein-energy malnutrition: effect of nutritional rehabilitation

A standard sleep questionnaire was given to the parents of 26 infants with protein-energy malnutrition who underwent polysomnographic evaluation. These investigations were repeated approximately 2 months after enrolment in a nutritional rehabilitation programme based on World Health Organization guidelines. Anthropometric values and serum serotonin levels were also measured. After nutritional rehabilitation there was a significantly higher percentage of non-rapid eye movement [REM] sleep; 2nd REM time, and latency times for sleep and REM sleep increased. Percentages of REM sleep and serum serotonin levels decreased significantly. Protein-energy malnutrition seems to affect the sleep-wake cycle; disturbed serotonin levels may be among the factors responsible

16S rDNA analysis for characterization of a bacillus isolate and its tolerance profile after subsequent subculturing

The isolation of microorganisms from different highly contaminated environments offers novel bacteria of unique functionality and potential applications in different biotechnological processes. It also entails a variety of genes responsible for microbial tolerance or defense against extreme conditions or xenobiotics present in the media. In a previous work, Bacillus spp. isolated from textile wastewater was exploited in terms of characterization, tolerance to pH, salinity, cold temperature and hydrogen peroxide, and the mechanism of resistance against hydrogen peroxide [Gomaa and Momtaz, 2006]. This isolate was phenotypically identified as Bacillus maroccanus/Bacillus simplex. In this study, the primary aim was to characterize this strain using 16S rRNA partial gene amplification and sequencing. It was assigned as Bacillus simplex TWW-04. DNA-DNA hybridization showed 96% relatedness to Acinetobacter. The tolerance profile of this strain was examined in regard to the four traits previously mentioned. A 36-month subculturing of this strain showed no change in tolerance to pH, salinity and cold temperature, whereas the tolerance to hydrogen peroxide was lost. A sample, frozen for the same duration, showed no change in any of the tested traits. The tolerance alleviation was evident after 24 months of subculturing. Testing the KatA gene responsible for hydrogen peroxide tolerance in the two samples showed higher band intensity in the frozen sample when compared to the subcultured sample. This indicates that the hydrogen peroxide tolerance trait is extrachromosomal and has been lost during successive cell division, while the other traits are chromosomal

Multiplex PCR assay for the simultaneous species identification and detection of antibiotic resistance genes in staphylococci isolates

The staphylococci are among the most prominent of all nosocomial pathogens. The progressive emergence and rapid dissemination of antibiotic resistance in staphylococci and its association with the use and consumption of antibiotics constitute a major health concern and have been considered a global crisis. In this study a multiplex PCR assay was performed to distinguish Staphylococcus aureus from the less virulent but clinically important coagulase-negative staphylococci [CoNS] and to simultaneously determine their antibiotic resistance profiles. Twenty staphylococci clinical isolates from hospital acquired infections were tested by the disk diffusion method and with multiplex PCR assays for species identification and to detect antibiotic resistance genes simultaneously in one PCR amplification. The targeted genes are implicated in resistance to oxacillin [mecA], gentamicin [aac[6']-aph[2"]], and erythromycin [ermA and ermC]. An additional primer pair for the amplification of a fragment of the staphylococcal 16S rDNA was included as a positive control. Another primer pair targeting an S. aureus-specific DNA fragment was included to facilitate the definite identification of S. aureus isolates. We found the correlation between genotypic and phenotypic analysis as follows: 93% for oxacillin resistance, 100% for gentamicin resistance, 80% for erythromycin resistance and 100% for staphylococcal species identification. The 10 staphylococci isolates that were resistant to gentamicin harbored the aac [6']-aph [2"] gene. Of 15 oxacillin-resistant staphylococci isolates, 14 [93%] were harboring mecA based on PCR analysis. Regarding the 15 staphylococci isolates that were resistant to erythromycin, there were 8 [53%] harboring one of the two erythromycin resistance gene tested and 3 [20%] harboring both genes while 4 [27%] were not harboring any of the genes. The isolates showing discrepant results between the genotypic and phenotypic methods should be further tested for another mechanism of resistance. The multiplex PCR assay offers a rapid and simple identification of antibiotic resistance profiles and may be used as diagnostic tool to replace or complement standard methods of susceptibility testing and identification of staphylococci

Obesity: a risk factor for acute myocardial infarction with angiographically patent epicardial coronary vessels in an adolescent

Toreport a causal relationship between myocardial infarction [MI] and obesity in an adolescent in the absence of the well-known risk factors for MI. A morbidly obese 17-year-old male, a nonsmoker, nondiabetic and normotensive patient, who sustained acute inferior MI with no family history of coronary artery disease, presented with central chest pain. ECG showed low voltage, normal sinus rhythm and ST segment elevation in the inferior leads; cardiac enzymes were elevated. Screening for ethanol and cocaine were negative. He was admitted to the coronary-care unit as a case of inferior MI with late presentation. Cardiac catheterization revealed patent epicardial coronary arteries; short- and long-term plans for weight reduction and family counseling were started. The hospital stay was uneventful, and the patient was discharged home on the fourth day. Based on clinical and laboratory findings, we assume that the MI might partly be secondary to coronary artery spasm or invisible premature atherosclerotic plaques. Public education and awareness for this complication in a young obese patient are warranted

Oxidative stress in preteclampsia: Role of homocysteine

To evaluate the possible role of homocysteine [Hcy] in inducing oxidative stress in preeclampsia, plasma Hcy was measured. Twenty preeclamptic pregnant, 20 normotensive pregnant and 20 healthy non- pregnant women were analyzed with regard to plasma Hcy, lipid peroxidation, antioxidant status, tumor necrosis factor-alpha [TNF- alpha], cholesterol, triglycerides, uric acid and creatinine. Plasma Hcy concentrations were significantly higher in preeclamptic women than in controls [11.9 and 8.3 mumol/L, respectively. Similarly, thiobarbituric acid-reactive substances [TBARS], as an index of lipid peroxidation were significantly higher in preeclamptic women than healthy non-pregnant women [1.97 vs 0.61 mumol/L, again with no significant difference between normotensive pregnant and healthy non- pregnant women. A significant decrease of antioxidant vitamins [vitamin C and vitamin E], glutathione [GSH] and antioxidant enzymes [superoxide dismutase [SOD], catalase and glutathione peroxidase [GSHPx]] were observed in the serum and erythrocytes of preeclamptic women compared to women with normotensive pregnancy. Serum TNF-alpha, cholesterol, triglycerides, uric acid and creatinine showed high levels in preeclamptic pregnant women when compared to both other groups

proximally and distally based peroneal artery flap: an antomical and clinical study

An anatomical study was carried out on 20 fresh and preserved cadaveric lower limbs after injection of the femoral artery by red latex. Muscular branches were given to the adjacent muscles and most of them continued as musculocutaneous arteries. Septocutaneous branches, 3-6, were given. A clinical study was also carried out on 20 patients suffered from traumatic tissue defects and exposed tibia, aged between 12 and 47 years. They were equally divided into two groups [A and B]. Their defects were treated using the proximally-and distally-based flaps sequentially. Two of the proximally-based flaps showed delayed necrosis of their distal ends [1.5 and 2 cm] that healed by conventional dressings. Skin grafting over the donor site was excellent in all cases of group A. One of the distally-based flaps showed delayed necrosis of its distal one-fourth that needed split thickness skin grafting. Another case of the same group [B] showed a partial loss of the immediate mesh skin grafting over the donor site which was healed by conventional dressings. Necrosis of the distal one-fourth of the distally-based flap may be explained by the anatomical fact that the lower third of the leg contains peroneal perforators with the least diameter and length

Serum selenium lipids abnormalities in cirrhotic patients with and without ischemic heart disease

Although the cirrhotic patients are less prone to develop ischemic heart disease [IHD], this study was done to investigate why some and rare cirrhotic patients develop this cardiac disease. Forty male cirrhotic patients of Child's B and C grading were selected twenty of them had only liver cirrhosis [group I] and the other twenty had also IHD [group II]. Liver function tests, serum lipid parameters and serum selenium were done for all subjects. ECG, echo - Doppler study and serum enzymes as CPK and LDH were done additional for patients of group II.Contrary to control, the cirrhotic patients showed highly significant reduction in serum selenium and HDL levels [P<0.01]. They also showed highly significant increases in total lipid, total cholesterol, LDL and triglyceride levels in the serum [P<0.01]. Contrary to the results of group I, serum results of group II showed significant reduction in selenium level, highly significant reduction in HDL and highly Significant increases in total lipids, total cholesterol, LDL and triglyceride levels.In both groups, serum selenium reduction was significant directly correlated with serum albumin level and was highly significant directly correlated with HDL level. However, it was highly significant inversely correlated with serum levels of total lipids, total cholesterol, LDL and triglyceride.It can be concluded that the cirrhotic patients had highly significant reduction in serum selenium and cardioprotective lipoprotein [HDL] levels. Also, they developed highly significant increases in the levels of atherogenesis-induced parameters as total lipids, total cholesterol, LDL and triglyceride. Thus, the highly significant hyperlipidaemia and atherogenesis in patients of group II may be attributed to more significant reduction in their serum selenium

Role of toxocariasis in some allergic skin diseases

Allergic skin diseases are cutaneous reaction attributed to multiple potential causes. One of these causes is toxocaral infection. In order to explore its role we examined two groups of children suffering from atopic dermatitis [AD] [26 children aging up to 15 years Group I] and chronic idiopathic urticaria [CIU] 24 children of the same age [Group II]. In addition to 30 apparently healthy children free from any skin disease, they were considered as control group [Group III]. Antitoxocaral IgG antibodies detected by ELISA were found among 15%. 21% and 10% of the three groups respectively with a statistical significant difference in the chronic idiopathic urticarial group [CIU] and the level of antitoxocaral IgG was significantly higher among the two groups versus the control one [P<0.05]. Specific antitoxocaral IgE detected by immunosorbent agglutination assay [ISAGA] revealed that, positivities to specific IgE were 7.7% and 12.5% in the atopic dermatitis and chronic idiopathic urticarial groups respectively with a high statistical significant difference versus the control group [P<0.001]. Estimation of eosinophils among the different groups revealed that, eosinophilia was found among the examined groups with a highly staistical significant difference than the control [P<0.001] and hypereosinophilia was present in patients showed positivity for IgE specific for toxocaral infection. It could be concluded that, toxocaral infection most probably has a role in some idiopathic skin allergic diseases as atopic dermatitis and chronic idiopathic urticaria

Oxidant/antioxidant status in children with bronchial asthma

Antioxidant-oxidant imbalances are thought to contribute to oxidative stress in bronchial asthma. The aim of this study was to examine serum malondialdehyde as a biomarker of oxidative stress in asthma in relation to dietary and metal scavengers antioxidants. In this study, serum levels of beta-carotene, vitamin A, alpha-tocopherol and ascorbic acid addition to serum albumin, transferrin, ceruloplasmin and malondialdehyde were investigated in 49 asthmatic children during an acute exacerbation and remission. 24 healthy children served as controls. Malondialdehyde levels were significantly higher at asthma attack compared to remission [P=<0.001]. Ascorbic acid, albumin and transferrin were significantly lower in asthmatics at remission compared to controls [P=<0.001] with no significant differences in their levels during acute exacerbation and remission periods. No differences were observed in the serum levels of beta-carotene, vitamin. A, alpha-tocopherol or ceruloplasmin between the groups. Although the mechanisms underlying the association between asthma and oxidant/antioxidant system are unclear, according to our findings, increased oxidant stress as reflected by malondialdehyde level and decreased ascorbic acid, albumin and transferrin antioxidant protection may contribute to the pathogenesis of asthma

Plasma levels of leptin in patients with chronic heart failure

Chronic heart failure [CHF] is a syndrome characterized by energy dysregulation. In this study, plasma levels of leptin, a protein hormone secreted by adipocytcs and involved in energy regulation, were determined in fourty patients with CHF divided into two equal groups; mild CHF patients, with New York Heart Association [NYHA] functional class 2, and severe CHF patients with NYHA functional class 3 or 4. Left ventricular ejection fraction [LVEF] as determined by echocardiography was < 45% in all patients. The study included also 20 healthy subjects matched for age, gender and body mass index, as controls. Plasma leptin levels were determined by radioimmunoassay. showed significant elevation in plasma leptin levels in patients with severe CHF [8.3 +/- 0.5 ng/nil] compared to controls [5.2 +/- 0.4 ng/ ml, p < 0.001] and mild CHF patients [6.8 +/- 0.6 ng / ml, p< 0.05], with a significant difference between mild CHF patients and controls [p < 0.05]. Signifcant negative [r = 0.362, p < 0.05] and positive [r = + 0.376, p < 0.05] correlations were noticed between plasma leptin levels and each of LVEF and NYHA functional class respectively. we found that patients with CHF were hyperleptinenic with significant correlation between plasma leptin levels and severity of the disease. Further studies arc recommended to clarify the significance of these findings, and we wonder whether leptin could be a marker and / or mediator of energy dysregulation in CHF patients

Myosin light chain one [M.L.C.I], a diagnostic marker for acute myocardial infarction [A.M.I.]

Myosin light chain one [MLCI], total CPK and total LDH were estimated in 70 individuals; 40 patients with acute myocardial infarction [AMI]; [18 patients were successfully reperfused with streptokinase [STK] while the remaining 22 patients received conventional treatment] and 30 healthy persons as a normal control group. All patients were followed up by serial ECG. MLCI was measured by enzyme immunoassay reagents supplied by Yamasa Shoyu Co. Ltd. Tokyo, Japan, while total CPK and LDH were determined Colorimetrically by reagents supplied by Boehringer Mannheim GmbH Mannheim, Germany MLCI was of value in the diagnosis of AMI. MLCI persisted for 10 days, that is more than any enzyme and it is of value to make the diagnosis in patients who presented late i.e. after one week of the infarction in whom the ECG is equivocal and all enzymes returned to the normal level. Both MLCI and CPK were positive in 11% while LDH was positive in 2.7% of cases after 8hrs of the onset of chest pain [OCP]. This positivity went up to 88% for MLCI, 100% for CPK and 44% for LDH 24hrs after OCP in patients who had conventional treatment only. In patients who received STK, the positivity for these enzymes was 50%, 100% and 44% respectively 24hrs after OCP

Delayed sternal closure for life threatening postoperative complications

In four years period, delayed sternal closure was decided in 22 patients. The main indications were uncontrollable bleeding [45.4%] and cardiac dilatation [54.6%]. In adults skin approximation was possible while young children synthetic membranes were used to cover the skin defect. Delayed sternal closure was possible after 24 hours in cases of bleeding and after 2-7 days in cases of cardiac edema. Three patients died before sternal closure and two died late in the postoperative period. The survival rate was 77.2%. The incidence of mediastinitis was 4.5%

Effect of aprindine and amiodarone administration on the incidence of reperfusion ventricular fibrillation in patients undergoing valve replacement

This prospective study was planned to evaluate the effect of the antiarrythmic drugs; Aprindine and Amiodarone on the incidence of reperfusion VF. 90 patients undergoing valve replacement were divided into control, aprindine and amiodarone groups. The incidence of VF decreased from a mean of 80% in the control group to 43.3% and 66.7% in the aprindine and amiodarone groups. The difference was statistically significant in the aprindine group. For defibrillation, a mean of 33.44 joules and 2.87 shocks was required in the control group, while the aprindine group required 8.33 joules and 1.33 shocks, and the amiodarone group required 13.84 joules and 1.77 shocks. Postoperative creatine kinase isoenzyme was 88.33 units in the control group and 53.55 and 58.80 units in the aprindine and amiodarone groups respectively. The incidence of reperfusion VF increased in patients receiving digitalis and in cases with ischemic time >90 minutes. Moreover, the incidence was higher, if the serum potassium was <5 mil Equ. or the body temperature was <34C, at time of aortic decalamping

use of fashioned pericardial cusps for repair of tricuspid endocarditis

A fashioned pericardial cusps and chordae were used for repair of tricuspid endocarditis. The operation was done for 4 patients [1 female and 3 males], their ages ranged from 16 to 35 years. Two of them developed the endocarditis as a complication after open heart surgery. The third one was drug abuser and the lady had the endocarditis after child-birth. All had +ve blood cultures, which could be controlled medically in three. The causative organisms were mixed in three patients. Grade 4/4 tricuspid incompetence was present in two, and 3/4 in the other two. The pulmonary artery pressure ranged from 38 to 55 mmHg. At operation, mitral valve replacement was done for two patients. Pericardial cusp replaced the septal leaflet in one patient, and the anterior leaflet, in two patients. The three leaflets were excised and replaced by two pericardial cusps in one patient. Vegetations were present in two patients only. Follow up for 1-3 years revealed [1-2]/4 tricuspid incompetence in one patient and trivial incompetence in the others

bacterial flora of carious lesions and caries-free surfaces in school students 13-16 years of age in Cairo

This study showed that the difference between group 3 and either of group 1 or group 2 was statistically significant as regards total bacterial count, total streptococcal count and Strep. mutans count. While, this difference was not significant between group 1 and group 2. It was also found that the difference in the mean percentage of total streptococcal count between the three groups was statistically insignificant at 5% level. As regards Strep. mutans, the difference in the mean percentage between group 1 and group 2 was not statistically significant; while it was significant between group 3 and either of group 1 or group 2 at 5% level

Study of the action of insulin-like growth factor [IGF-I] on the transfer and uptake of nutrients in the human placenta perfused in vitro

The aim of the present study is to investigate the effect of IGF-I on the transfer and uptake of glucose, lactate and AIB in the human placenta. IGF-I [5ng/ ml] caused an increase in glucose transfer after 20 minutes of its addition, then it decreased gradually till it reached its minimal value after 35 minutes. On the other hand, there is no significant change in glucose uptake. There is no significant change in both transfer and uptake of alpha -aminoisobutyric acid [AIB] through the placenta after the addition of 5ng/ml IGF-I.IGF-I caused a slightly diminition in fetal lactate release and a moderate decrease in the maternal lactate release. The reasons for such results might relate to the difference in gestational age of the placental tissues examined and the short period of observation presented