Serum pattern of circulating soluble receptor of leptin throughout the menstrual cycle.

AIM: The aim of the study was to investigate the pattern of circulating soluble receptor of leptin (sLeptinR) during the menstrual cycle and the association of sLeptinR to leptin, sex hormones and gonadotropins. METHODS: Fasting blood samples were collected on alternate days throughout a full cycle from fifteen healthy volunteers. Immunoenzymatic assays were employed to record the relevant levels. RESULTS: sLeptinR concentrations throughout the cycle were found to vary negligibly. No significant correlations between sLeptinR and leptin, gonadotropins or progesterone, were established. During the follicular phase, subjects presenting with higher estradiol levels tended to have higher sLeptinR concentrations. CONCLUSION: The little variation of sLeptinR concentrations during the menstrual cycle indicates that the rise of leptin during the luteal phase implies an increase of its bioactivity.

Increased seroprevalence of Toxoplasma gondii in a population of patients with Bell's palsy: a sceptical interpretation of the results regarding the pathogenesis of facial nerve palsy.

Facial nerve oedema and anatomical predisposition to compression within the fallopian tube seem to be the only generally accepted facts in the pathophysiology of Bell's palsy. Several infectious causes have been suggested as possible triggers of this oedema. Most of the suggested pathogens have been associated with facial nerve lesions during latent infections, reinfections or endogenous reactivations. The aim of this study was to investigate the seroprevalence of three such pathogens Toxoplasma gondii, Epstein-Barr virus (EBV) and cytomegalovirus (CMV) in a population of patients with facial nerve palsy. Fifty-six patients with Bell's palsy were included in the study. A group of 25 individuals with similar age and gender distribution was used as control. Seropositivity for T. gondii, EBV viral capsid antigen (VCA) and CMV-specific IgM and IgG antibodies was investigated 2-5 days after the onset of the palsy. Comparisons for both IgM and IgG antibodies against T. gondii attributed significantly higher seroprevalence in the patients' group than in the control group (p = 0.024 and 0.013, respectively). The respective examinations for EBV and CMV attributed no significant results. The roles of EBV and CMV in the pathogenesis of Bell's palsy were not confirmed by this study. However, a significantly higher seroprevalence of IgM- and IgG-specific T. gondii antibodies was detected in patients with Bell's palsy when compared to healthy controls. The possibility that facial nerve palsy might be a late complication of acquired toxoplasmosis may need to be addressed in further studies.

The role of breastfeeding and passive smoking on the development of severe bronchiolitis in infants.

AIM: Bronchiolitis is an acute infectious disease of the lower respiratory tract which causes the obstruction of bronchioles in children younger than 2 years. The aim of this study was to investigate the effect of passive smoking alone and in conjunction with breastfeeding on the severity of acute bronchiolitis in infancy and the duration of hospitalisation. METHODS: We studied 240 consecutive infants aged from 6 to 24 months (137 boys and 103 girls) median age 14 months, who required hospital admission for acute bronchiolitis at the Paediatric Department of Democritus University Hospital, Alexandroupolis, Greece. The outcomes of interest were the severity of bronchiolitis and the duration of hospitalisation. RESULTS: Among the entire cohort, 122 (50.8%) children presented a severe attack of bronchiolitis. In multivariate regression analysis adjusting for confounding factors, breastfeeding for less than four months (aOR=6.1, 95% CI=3.4-10.7), exposure to environmental tobacco smoke (aOR=2.2, 95% CI=1.1-3.6) and their combination (aOR=16.2, 95% CI=6.0-34.3) showed significant association with severe bronchiolitis and prolonged hospitalisation. Passive smoking did not increase the risk of severe bronchiolitis, when infants breastfed for more than four months (aOR=1.9, 95% CI=0.8-5.1). CONCLUSION: In conclusion, exposure to environmental tobacco smoke worsens the symptoms and the prognosis of bronchiolitis, while breastfeeding seems to have a protective effect even in children exposed to environmental tobacco smoke.

Diagnostic usefulness of bone marrow aspiration material for the amplification of IS6110 insertion element in extrapulmonary tuberculosis: comparison of two PCR techniques.

SETTING: In many cases of extra-pulmonary tuberculosis (EPTB), with the exception of paucibacillary analysed specimens, the suspected site of mycobacterial infection is relatively inaccessible or unknown, making laboratory confirmation of TB laborious and problematic. OBJECTIVE: Two different polymerase chain reaction (PCR) based methods were compared to investigate the validity of bone marrow aspiration material as an easily accessible alternative sample for molecular analysis in EPTB. DESIGN: We amplified the same sequence of IS6110 of Mycobacterium tuberculosis complex in 19 confirmed cases of EPTB using two different nested PCR techniques: one in-house 'classic' PCR and another based on LightCycler technology. RESULTS: Both methods demonstrated the same reliability when performed in samples of infected tissue. However, the LightCycler protocol was superior to the in-house system when applied in bone marrow aspiration material, revealing positivity in 18/19 compared to 13/19 samples of 'classic' PCR. CONCLUSION: The application of an optimised LightCycler nested amplification protocol in bone marrow aspirates may promote diagnostic accuracy in difficult and/or urgent cases of EPTB.

Nosocomial lower respiratory tract infections: prevalence and risk factors in 14 Greek hospitals.

Nosocomial lower respiratory tract infections (NLRTIs) are associated with significant morbidity and mortality. The aim of this study was to investigate the epidemiological features of NLRTIs in Greece, where knowledge about these infections is limited. Two point-prevalence studies of hospital-acquired infections were carried out in 14 Greek hospitals located throughout the country, one in 1999 and one in 2000. NLRTIs were diagnosed in accordance with the Centers for Disease Control and Prevention (CDC) definitions. Among the 7,120 hospitalized patients registered during the two studies, 610 (8.6%) cases of hospital-acquired infections were identified, of which 200 (32.8%) were NLRTIs. Sixty-nine (34.5%) patients had pneumonia, and the remaining 131 (65.5%) patients had bronchitis. The greatest prevalence of NLRTI was found in the adult ICUs (30.4%). Male gender, age >65 years, mechanical ventilation, tracheostomy, an intravenous central line, and an indwelling urethral catheter were the main risk factors. There was no significant difference in the incidence of NLRTI among hospital-acquired infections between the 1999 study and the 2000 study. The causative microorganism was identified in 78 of 200 (39%) cases, and 103 strains were isolated. The majority of strains (67%) were gram-negative bacteria. The most frequently isolated microorganisms were Pseudomonas aeruginosa (22.3%), Acinetobacter spp. (19.4%), Klebsiella pneumoniae (12.6%), and Staphylococcus aureus (10.7%). There was no difference between the two prevalence studies in the frequency of isolation of the microorganisms. NLRTI was the leading cause of morbidity and mortality among hospitalized patients with hospital-acquired infections in Greek hospitals. Gram-negative microorganisms were the most frequently isolated pathogens.

Relationship between nosocomial Acinetobacter species occurring in two geographical areas (Greece and the UK).

Fifty-two isolates of Acinetobacter spp. obtained from three Greek and one UK hospital, were studied using partial 16 S ribosomal DNA sequence analysis, repetitive extragenic palindromic sequence-based polymerase chain reaction (REP-PCR) mediated fingerprinting and DNA macro-restriction analysis. The aim was twofold: first, to discern the major differences in the population of Acinetobacter spp. between the two countries. Second, to compare a simple PCR-based typing scheme with pulsed-field gel electrophoresis (PFGE). The multi-resistant Greek isolates were within DNA groups 2 and TU13, and clustered into three types both by REP-PCR and PFGE. By contrast, the more susceptible Oxford isolates were heterogeneous on 16 S RNA sequence analysis and distinguishable on typing. The need for studies that elucidate the phylogeny of Acinetobacter spp. inside and outside hospitals are important, as this will help clarify the relationship between organisms that are increasingly recognized as causes of severe infections.

Tumour necrosis factor-alpha levels in non-diabetic offspring of patients with type 2 diabetes mellitus.

Tumour necrosis factor-alpha (TNF-alpha) is considered to be involved in the insulin resistance of type 2 diabetes mellitus. The offspring of patients with type 2 diabetes mellitus are at increased risk of developing diabetes and several metabolic abnormalities, but the underlying defects responsible are not known. We studied serum TNF-alpha levels in 30 healthy non-diabetic offspring of type 2 diabetic parents (group A), and the relationship between TNF-alpha levels and variables associated with insulin resistance and diabetes. For comparison, 30 healthy offspring of non-diabetic parents (group B) were also studied. The median serum concentration of TNF-alpha was significantly higher in group A than in group B, 3.5 pg/ml compared with 2.0 pg/ml, respectively. The individuals of group A also had significantly elevated levels of glycosylated haemoglobin, fasting glucose, glucose 2 h after an oral glucose tolerance test and triglycerides. We conclude that serum TNF-alpha concentration is significantly elevated in non-diabetic offspring of type 2 diabetics and this may predict later impairment of insulin action in these individuals.

Amplification of IS6110 sequence for detection of Mycobacterium tuberculosis complex in HIV-negative patients with fever of unknown origin (FUO) and evidence of extrapulmonary disease.

OBJECTIVES: Extrapulmonary tuberculosis (TB) constitutes the main cause of classic fever of unknown origin (FUO) in many populations. The aim of this study was to improve the diagnostic field of the disease using a nested polymerase chain reaction (PCR) assay, specific for the IS6110 insertion element of Mycobacterium tuberculosis complex, in order to achieve a more timely diagnosis and treatment. SETTING: Twenty-four, HIV-negative classic FUO patients who were admitted to the Regional Hospital of Alexandroupolis between April 1997 and July 1999. SUBJECTS AND DESIGN: The above patients were considered as putative extrapulmonary TB after 3 weeks of in-patient investigation and underwent exhaustive examination for diagnosis of the disease. For this purpose, specimens were obtained from peripheral blood and bone marrow from these patients, as well as from damaged tissues, and analysed by both PCR and conventional methods. Anti-tuberculous treatment was initiated in 16 out of 24 patients and the response to this regimen was considered as the final criterion for diagnosis of tuberculosis. RESULTS: Extrapulmonary TB was established in 11 patients. The PCR-based methodology, when applied to samples derived from bone marrow aspirations and suspected damaged tissues, was able to diagnose 10 of them, whereas the conventional methods were able to detect only two. CONCLUSIONS: Our results confirm the diagnostic value of molecular detection of M. tuberculosis in cases of FUO, thus supporting the application of PCR in tissue samples suspected of bacillus infection. Furthermore, our studies demonstrate that bone marrow aspiration specimens constitute an alternative, easy, safe and reliable source for such PCR analysis.

Evaluation of changes in serum and dialysate levels of cancer antigen 125 in stable continuous ambulatory peritoneal dialysis patients.

To estimate the relationship between changes in the concentration of cancer antigen 125 (CA125) and peritoneal membrane kinetics, the permeability characteristics of 44 continuous ambulatory peritoneal dialysis (CAPD) patients who had been treated with peritoneal dialysis for at least six months were prospectively evaluated. Twenty-seven males (age 66 +/- 6 years, duration of CAPD 35.5 +/- 29 months) and seventeen females (age 63.7 +/- 9 years, duration of CAPD 47.7 +/- 32 months) were evaluated. Peritoneal equilibration test (PET) data and Adequest (Baxter Healthcare Corporation, Deerfield, Illinois, U.S.A.) data were analyzed in all patients over a 12-month period, while CA125 levels were measured in blood and dialysate samples. No statistically significant correlations were seen between the patients' age, sex, or peritonitis incidence rates, and serum and dialysate levels of CA125. Dialysate-to-plasma ratio (D/P) of small solutes at 0 and 240 minutes also showed no statistical correlation. Statistical analysis revealed a statistically significant negative correlation (r = -0.33, p = 0.035) between dialysate CA125 and duration of CAPD. The statistically significant difference found between dialysate CA125 concentrations at 0 minutes and 240 minutes (2.32 +/- 1.3 U/mL vs 9.08 +/- 6.8 U/mL, p < 0.0001), means that CA125 concentration increases with longer dwell time. These results suggest that the duration of CAPD clearly affects dialysate CA125 concentrations. CA125 may therefore be used as a useful marker to evaluate the mesothelial cell mass in longitudinal follow-up.

Treatment and prevention of relapses of CAPD Pseudomonas peritonitis.

Pseudomonas peritonitis in continuous ambulatory peritoneal dialysis (CAPD) can be difficult to eradicate, because it is frequently resistant to common antibiotics, inducing the loss of the peritoneal cavity in some cases. A total of 14 episodes of Pseudomonas peritonitis in 12 patients (6 male, 6 female) were treated with intraperitoneal (IP) administration of a combination of ceftazidime and tobramycin. All patients were hospitalized. The loading doses were 1000 mg/2 L of ceftazidime and 1.7 mg/kg of tobramycin, and the maintenance IP doses were 250 mg/2 L of ceftazidime and 16 mg/2 L of tobramycin. The therapy duration was 14 days. In 7 episodes (group A) no other antibiotic regimen was provided, while in the remaining 7 episodes (group B) therapy was continued with 500 mg b.i.d. of oral ciprofloxacin for the next 14 days. Pseudomonas species isolated in group A were P. alcaligenis (1), P. putida (1), P. maltophilia (1), R. cepacia (1), and unidentified (3). In group B the following Pseudomonas species were isolated: P. aeruginosa (4), P. diminuta (1), P. stutszeri (1), and unidentified (1). Recurrence of peritonitis was seen in 4 episodes of group A with 2 catheter removals, while all episodes were cured in group B. These results suggest that IP ceftazidime and tobramycin with the additional use of oral ciprofloxacin is successful in the treatment and prevention of relapses of Pseudomonas peritonitis.

Treatment of CAPD peritonitis with clavulanate potentiated ticarcillin.

A total of 16 episodes of peritonitis in 14 patients (9 males, 5 females), were treated with Clavulanate potentiated ticarcillin (TC), a -lactamase stable parenteral penicillin. All the pts were hospitalized and received initial loading dose of 3.2 gr intraperitoneally (i.p.) in a 6-hour 1 L exchange, which was followed by four 1 L exchanges with 320 mg/LTC. The therapy was continued for ten days. The bacteria isolated were: Staph. epid. (4), Staph. aureus (2), Strept. viridans (1), Enterococcus (1), Klebsiella Pneum. (1), Serratia (1), Enterobacter (1), Pseudomonas species: stutszeri (2), cepacia (1), fluorescens (1), negative cultures (1). Recurrence of peritonitis was seen in three patients with Pseudomonas (stutszeri (2), fluorescens (1)) peritonitis, 10-16 days after cessation of therapy. No clinical or biological side effects were seen in any patient during and/or after the therapy. These results suggest that, i.p. monotherapy of TC is effective in the treatment of CAPD peritonitis, while in cases of Pseudomonas peritonitis more specific regimens should be used.

Vancomycin therapy for gram-positive peritonitis in patients on CAPD.

We describe the use of vancomycin in the therapy of gram-positive peritonitis in patients on CAPD. Two ways of administration were in comparison: a) the intraperitoneal (IP) route, with the intraperitoneal administration of 30 mg/l vancomycin for 10 days and b) the intravenous (IV) route with 2 infusions of 1 gram of vancomycin, the first one on the day of the diagnosis of gram-positive peritonitis and the second 7 days later. Each one of these therapeutic schedules was applied at random for 20 episodes of peritonitis out of 40 episodes with gram-positive organisms (28 Staph. albus, 10 Staph aureus and 2 Streptococcus). Remission of clinical symptoms occurred in 49-72 hours in both groups, while macroscopic clearing of dialysate effluent and sterilization of cultures were observed in 4-7 days. Recurrence of peritonitis was seen in 4 patients of the IV group 2 weeks after the administration of the second dose of vancomycin. We conclude that the use of vancomycin with either of the two ways of administration is of great value in the treatment of gram-positive peritonitis. The IV infusion was less successful but it seems to be useful for the out-patient treatment of gram-positive peritonitis.