Fast and specific dermatophyte detection by automated DNA extraction and real-time PCR.

The aim of this study was to develop and validate a rapid and sensitive real-time PCR method for detection of all known species of dermatophytes, including identification of Trichophyton rubrum and Trichophyton interdigitale. Fungal DNA was extracted directly from clinical samples by using a pre-lysis step, followed by automated DNA extraction on the MagNA Pure Compact. In total, 202 clinical samples were examined by both conventional culture and by the new PCR method. In 103 (51%) of the samples fungal nucleic acid was detected by PCR, while only 79 (39%) were found to be positive by culture. Out of 103 PCR-positive clinical samples, 94 (91%) were identified as T. rubrum and eight (8%) as T. interdigitale. This real-time PCR is far more sensitive and 2-4 weeks faster than conventional culture for detection of dermatophytes present in clinical samples.

Housekeeping health care workers have the highest risk for tuberculin skin test conversion.

BACKGROUND: Not all health care workers (HCWs) are at the same risk for tuberculin skin test (TST) conversion, indicating latent tuberculosis (TB) infection. OBJECTIVE: To identify risk factors for TST conversion among HCWs. METHODS: A retrospective cohort study conducted at a tertiary university medical centre included every HCW who had had a negative two-step TST at work entry and at least one consecutive TST in the period 2005-2009 (mean follow-up period 55 months). Binomic logistic regression analysis was used to identify risk factors for TST conversion. Potential risk factors such as age, health care profession, patient exposure profile, workplace division and history of bacille Calmette-Guérin vaccination were entered in the model. RESULTS: A total of 450 subjects met the inclusion criteria, of whom 93 had TST conversion. The highest annual rates of TST conversion occurred in workers who worked as housekeeping staff (6.9%). Older age, a work environment with high patient turnover and employment in maintenance departments were significant risk factors (adjusted odds ratios 2.05, 5.2 and 8.4 respectively). CONCLUSION: Housekeeping staff, older age workers and health care professionals working in an environment of high patient turnover are at increased risk for latent TB infection.

Alarming spread of vancomycin resistant enterococci in Sweden since 2007.

The total number of persons infected or colonised with vancomycin-resistant enterococci mandatorily reported to the Swedish Institute for Infectious Disease Control increased dramatically during 2007 and 2008. During a period of twenty months from 1 July 2007 to 28 February 2009, a total of 760 cases were reported compared with 194 cases reported during the entire period from 2000 to 2006. This rise was mainly attributed to a wide dissemination of vancomycin resistant enterococci which started in a number of hospitals in Stockholm in the autumn of 2007 and was followed by dissemination in various healthcare facilities (hospitals and homes for the elderly) in a further two Swedish counties in 2008. The majority of the cases (97%) were acquired in Sweden and among these, healthcare-acquired E. faecium vanB dominated (n=634). The majority of these isolates had identical or closely related pulsed-field gel electrophoresis patterns indicating clonal dissemination in the affected counties. The median minimum inhibitory concentration of vancomycin was 32 mg/L (ranging from 4 to >128 mg/L) and of teichoplanin 0.12 mg/L (ranging from 0.06 to 0.25 mg/L). Particular emphasis was placed on countermeasures such as screening, contact tracing, cleaning procedures, education in accurate use of infection control practices as well as increasing awareness of hygiene among patients and visitors. With these measures the dissemination rate decreased substantially, but new infections with the E. faecium vanB strain were still detected.

Pleural effusion associated with rheumatoid arthritis: what cell predominance to anticipate?

Pleural involvement is the most frequent manifestation of rheumatoid arthritis (RA) in the chest. We report here two patients who presented with large exudative pleural effusions and subsequently developed sero-positive RA. In both cases, the differential cell count of the pleural effusion suggested empyema. A literature review identified that RA-associated pleural effusion afflicts more men than women and 95% of the patients have high titers of rheumatoid factor (RF). In 46% of cases, RA-associated pleural effusion is diagnosed in close temporal relationship with the diagnosis of RA. The effusion is an exudate and is characterized by low pH and glucose level, and high lactic dehydrogenase (LDH) and cell count. At diagnosis there is a tendency for predominant neutrophils to occur consistent with an empyema and 7-11 days later, the cells in the pleural effusion are replaced by lymphocytes. Pleural effusion with predominant eosinophilia is rare. RA patients with acidic effusion and low glucose content with neutrophils predominance should be treated with thoracic drainage and antibiotics until an infection is ruled out. The histo-pathologic findings in pleural fluid of tadpole cells and multinucleated giant cells and the replacement of the mesothelial cells on the parietal pleural surface with a palisade of macrophage derived cells are described as pathogonomic for RA. Treatment with systemic steroids and intra-pleural steroids are effective in most cases.

Can the routine use of fluoroquinolones for community-acquired pneumonia delay the diagnosis of tuberculosis? A salutary case of diagnostic delay in a pilgrim returning from Mecca.

Guidelines advocate using fluoroquinolones as first-line treatment for community-acquired pneumonia (CAP). However, the use of fluoroquinolones in patients with undiagnosed tuberculosis may cause a delay in the diagnosis of tuberculosis and may also promote the development of resistance to these drugs if used as monotherapy in undiagnosed tuberculosis. We illustrate the former with the following case report of a patient who developed tuberculosis after a pilgrimage to Mecca.

Montelukast treatment does not affect peripheral blood neutrophil functions in asthma patients.

BACKGROUND: Leukotriene receptor antagonists have become an integral part of asthma treatment as an add-on therapy for patients with moderate disease activity. OBJECTIVE: To determine whether in vivo treatment with montelukast, a leukotriene receptor antagonist, at the recommended therapeutic dose affects neutrophil functions which are essential for host defense. METHODS: Twenty nonsmoking patients between the ages of 18 and 70 with moderate asthma were recruited to the study. All of them were receiving inhaled corticosteroids for at least 6 months and beta2-agonist as needed. After the 2-week run-in period, in addition to the regular medication, each patient was treated with 10 mg of montelukast for a 6-week period. Pulmonary function test was performed and blood was tested for neutrophil activity twice during the run-in period and twice during the treatment period. Each assay was performed in parallel to a matched control. RESULTS: The 6-week treatment period with montelukast did not significantly affect neutrophil chemotaxis or phagocytosis nor the elevated superoxide production. However, at 2 weeks of treatment some of the patients showed a reduction in neutrophil function. CONCLUSIONS: The study demonstrated that the recommended dose of montelukast has no significant effect on peripheral blood neutrophil activities, indicating that montelukast treatment does not induce an inflammatory process and does not interfere with the first line of host defense.

Intradialytic hypercapnic respiratory failure managed by noninvasive assisted ventilation.

We report a hemodialysis patient with acute hypercapnic respiratory failure managed on noninvasive intermittent positive pressure ventilation and progressive metabolic acidosis. Dialysate bicarbonate concentration of 25 mEq/l was associated with exacerbation of metabolic acidosis, while higher dialysate bicarbonate concentration of 30 mEq/l induced a dangerous increase in PCO(2) level. Excessive bicarbonate buffering and CO(2) production induced by severe metabolic acidosis, malnourishment and tissue hypoxia, could explain inadequate correction of metabolic acidosis and worsening of hypercapnia in this patient. Our findings suggest the need for close monitoring of blood gases and cautious modulation of dialysate bicarbonate concentration in the presence of progressive metabolic acidosis in hypercapnic hemodialysis patients.

[Specialty clinics--gain or loss?].

We evaluated the significance of first visits to our pulmonary clinic with regard to diagnosis and treatment by reviewing records of 287 new referrals by medical care providers (Sept. 1998-Feb. 1999). At the first visit the diagnosis was changed in 30%, and treatment in 40%. These are minimal figures because evaluation had not been completed nor the diagnosis determined in all cases, while further investigation and follow-up continued. In light of these findings, we believe that recent pressure on primary care physicians to avoid referrals to specialty clinics would result in short-term savings, but in the long term, would increase expenses and diminish quality of care. It is important to consider ways to maximize the interaction between the primary care physician and the specialist to maintain quality of care and decreases costs.

Influence of asthma in pregnancy on labor and the newborn.

Asthma in pregnancy has been associated with maternal and fetal morbidity and mortality. This study examines the relations of asthma in pregnancy, its severity and its treatment to the labor process, maternal and fetal parameters. Hundred and one consecutive asthmatic women, who gave birth to single babies between November 1993 and November 1994 at the Soroka Medical Center were studied. A group of 77 nonasthmatic women, matched for age and ethnic origin, who gave birth to single babies during the same period served as controls. A larger percentage of asthmatic women suffered from respiratory and urinary tract infections than in the control group (p < 0.001). Severe asthma was associated with a higher rate of infections than milder asthma (p = 0.01). The incidence of smoking was higher among asthmatic women than among controls (p = 0.037). No association was found between socioeconomic status and smoking or infections. No association was found between maternal asthma or maternal use of corticosteroids and the following: maternal hypertension, maternal diabetes, low birth weight (< 2,500 g), preterm delivery (< 37 weeks), adequacy of weight to gestational age and Apgar scores. Three infants with congenital heart defects were born to asthmatic mothers. When the presentation of the fetus was not cephalic, all the asthmatic women were delivered by cesarean section, versus only 60% in the control group (p = 0.08). Labor was induced with oxytocin more often in the asthma group than in the control group (p = 0.07). We conclude that the labor and neonatal outcome in pregnant asthmatic women treated medically is good, even when asthma is severe and when the patient is treated with corticosteroids. There is, however, a relation between asthma in pregnancy, especially if severe, and predisposition to infections.

Sub-clinical worsening of bronchial asthma during estrogen replacement therapy in asthmatic post-menopausal women.

BACKGROUND: Changes in asthma activity, in part related to the female hormonal profile, have been observed during pre-menstrual periods and during pregnancy. Estrogen replacement therapy (ERT) is an accepted routine treatment for post-menopausal women. The effect of ERT on disease activity in post-menopausal asthmatic women has not been investigated in the past and is the subject of the present study. METHODS: Fifteen post-menopausal women with mild to moderate asthma completed two 30-day periods in which they measured peak expiratory flow (PEF) at home and filled in a daily diary of asthma-related symptoms. The first monitoring period was pre-ERT and the second was during ERT. In addition spirometry was performed on each woman three times, twice pre-ERT and once during ERT. RESULTS: The average daily PEF decreased from 241 (57.9, S.D.) l/min pre-ERT to 226.7 (62.7) l/min during ERT (P < 0.004). Significant differences between the two study periods were also found in morning and evening PEF values. Diurnal variation, measured as the difference between morning and evening PEF values, decreased significantly from 22.3 (26.7) l/min pre-ERT to 17.5 (26.8) l/min during ERT (P < 0.007). The average daily consumption of bronchodilator inhalers increased significantly from 3.7 puffs/day pre-ERT to 4.3 puffs/day during ERT (P < 0.006). Although the differences in spirometry between the two periods did not reach statistical significance, a trend towards a worsening of the obstructive disorder during ERT was observed. However, the general feeling of well-being of the asthmatics did not change during the two periods. CONCLUSIONS: During ERT a sub-clinical worsening of disease activity was found in postmenopausal women with mild to moderate asthma. We also detected a decrease in diurnal variation. Our findings should be substantiated by additional studies.

[Seasonal variation in hospital admissions for cardiorespiratory diseases in the Negev].

Characteristics of patients admitted to the general medical wards of this hospital during the 21 months between January 1990 and September 1991 were analyzed. Cardiovascular and respiratory disease-related (CR) admissions accounted for 46.8% of the 18,774 admissions. Using a linear model, we examined the relationship between season and clinical characteristics of the patients admitted with CR diagnoses. They were admitted more often during the winter (December-February) than the summer (June-August). The ratio of CR admissions to other causes of admission was 0.75 in the summer, 0.93 in the spring, 1.01 in the winter, and 0.82 in the autumn (p < 0.0001). Seasonality affected more patients with coronary heart disease, noncoronary cardiac disease, and chronic pulmonary disease than those with pneumonia or other pulmonary diseases, or cerebrovascular events. Those admitted during the winter were older, had more than 1 CR condition, and their hospital stays were longer. This pattern of hospitalization partly explains the increase in admissions to medical wards in the winter with increase in work load.

Influence of estrogen replacement therapy on airway reactivity.

The effect of estrogen on smooth muscle in various organs is unpredictable. Little is known about the effect of estrogen on respiratory tract smooth muscle, particularly in humans. In the present study we used the histamine challenge test (HCT) to assess the effect of estrogen replacement therapy (ERT) on airway reactivity in postmenopausal women who did not suffer from respiratory disease. Thirty-six women who were undergoing treatment at the postmenopausal clinic completed the study. All participants were nonsmokers whose pulmonary function tests were normal. HCT was performed twice before the inception of ERT, and a third time 4-6 weeks after ERT was begun. None of the 36 women demonstrated a 20% decrease in FEV1 values (PC20) after inhaling histamine at a concentration of 8 mg/ml, either before or during ERT. The maximal decrease in FEV1 values in response to the maximum concentration of histamine was significantly lower during ERT compared to the pretreatment period. The average maximal decrease in FEV1 during ERT was 2.63 +/- 2.72% (mean +/- DS) compared to 5.21 +/- 4.47% and 6.57 +/- 5.28% on the 2 tests prior to therapy (p < 0.0002). We conclude that ERT has an inhibitory effect on the bronchial reactivity of respiratory smooth muscle. There is no cause for concern about increased airway reactivity as an adverse effect of this therapy.

TGF-beta 1 causes increased endothelial ICAM-1 expression and lung injury.

Neutrophil adherence to vascular endothelium is partially mediated by adhesion molecules, including intracellular adhesion molecule 1 (ICAM-1), on endothelial cells. We examined the effect of transforming growth factor-beta 1 (TGF-beta 1) on the expression of ICAM-1 in human umbilical vein endothelial cells (HUVEC). TGF-beta 1 (1 ng/ml) increased ICAM-1 and ICAM-1 mRNA expression in HUVEC, as assessed by flow cytometry and Northern blot analysis, respectively. In addition, we investigated whether exogenous recombinant TGF-beta 1 can cause neutrophil-mediated lung injury in guinea pigs. The plasma half-life of 125I-labeled TGF-beta 1 in guinea pigs was 4.6 +/- 0.1 min, and the 125I activity was 2.8 +/- 0.2% 8 h after injection. The ratio of 125I-labeled albumin concentration in lung tissue and bronchoalveolar lavage (BAL) fluid to that in plasma, lung wet-to-dry weight ratio, numbers of neutrophils in BAL fluid, and numbers of neutrophils per alveolus in fixed lung sections increased in guinea pigs that received a high dose of TGF-beta 1 (25 micrograms i.v. followed by 2 micrograms/h for 8 h) compared with the control group. These results suggest that TGF-beta 1 causes neutrophil-mediated lung injury, possibly through upregulation of ICAM-1 on endothelial cells, and might be important in the pathogenesis of lung injury.

The protein kinase C inhibitor, H-7, induces acute lung injury in guinea pigs.

OBJECTIVES: To determine if the protein kinase C inhibitor, H-7, alone can cause acute lung injury. In cell studies, H-7 inhibited phorbol myristate acetate-induced neutrophil oxygen radical release. Additionally, one animal study demonstrated that H-7 inhibited phorbol myristate acetate-induced lung injury. There have been no studies on the effect of H-7 alone on lung function or on neutrophil release of oxygen radicals. DESIGN: Prospective, randomized, laboratory study along with in vitro studies using flow cytometry and lucigenin-dependent chemiluminescence. SETTING: Experimental laboratory. SUBJECTS: Specific, pathogen-free guinea pigs and isolated human peripheral neutrophils. INTERVENTIONS: Guinea pigs were randomized into three experimental groups: saline control, H-7 low dose (2 mg/kg bolus + 0.2 mg/kg/hr), and H-7 high dose (6 mg/kg bolus + 0.5 mg/kg/hr). Human neutrophils were randomized into control and experimental groups. The effects of H-7 on pulmonary permeability in guinea pigs were examined over an 8-hr period. MEASUREMENTS AND MAIN RESULTS: We measured the wet/dry weight ratio as an index of pulmonary edema and we measured the concentration ratios of 125I-labeled albumin in lung tissue and in bronchoalveolar lavage fluid and compared the ratios with those values in plasma as indices of pulmonary permeability. We also studied the in vitro effect of H-7 on human neutrophil oxygen radical production, using flow cytometry and lucigenin-dependent chemiluminescence. By flow cytometry, we measured oxygen radical production using the 2',7'-dichlorofluorescin and hydroethidine assays. The 2',7'-dichlorofluorescin assay mainly measures hydrogen peroxide, while the hydroethidine assay measures either superoxide anion alone or in combination with other oxygen intermediaries like hydrogen peroxide. Neutrophils (5 x 10(5)) were obtained by Ficoll-Hypaque gradient centrifugation and were incubated with H-7 (5, 25, 100 microM). In the H-7 high-dose group, wet/dry weight ratio, and 125I-labeled albumin ratios in lung/plasma, and bronchoalveolar lavage/plasma were significantly increased (p < .05 for each ratio). Pulmonary endothelial gap and subendothelial bleb formation were demonstrated in the high-dose group by electron microscopy. One hundred micromols of H-7 caused a small, significant decrease (23.3%, p < .05) in neutrophil oxygen radical production assessed by 2',7'-dichlorofluorescin. H-7 had no other effects on neutrophil oxygen radical production. H-7 did not stimulate neutrophil chemiluminescence; it decreased chemiluminescence. CONCLUSIONS: a) Protein kinase C inhibition with high-dose H-7 increased wet/dry weight and albumin in lung/plasma and bronchoalveolar lavage/plasma ratios in guinea pigs; b) the H-7 high-dose group demonstrated damaged pulmonary endothelium by electron microscopy; and c) since neutrophil oxygen radical production was not increased by H-7 as assessed by flow cytometry and chemiluminescence, it appears that H-7-induced acute lung injury and endothelial damage are not mediated by increased neutrophil oxygen radical production.

Perceptions of asthma severity: the role of negative affectivity.

The role of negative affectivity on patients' perceptions of and behavior during asthma attacks was explored among 47 asthmatic subjects. Patients completed 17 to 30 daily questionnaires assessing negative affect, asthma perception, additional drug intake, search for medical assistance, and peak-flow measures of respiratory distress. Asthma perceptions were correlated with negative affect and educational level; the perception of the asthma severity, but not negative affect, did predict behavior during an attack. The importance of symptom perceptions for the clinical management of chronic asthma is underscored.

Attenuation of acute lung injury and oxygen radical production by the 21-aminosteroid, U-78518F.

Oxygen radicals play an important role in the mechanism of acute lung injury. The 21-aminosteroid lazaroid, U-78518F, is a potent antioxidant. We examined the effect of intravenous U-78518F on acute lung injury in septic guinea pigs over 8 h. The experimental groups (n = 6) were 1) saline control, 2) Escherichia coli (2 x 10(9)/kg i.v.), 3) pretreatment (U-78518F 5 mg/kg bolus + 1 mg.kg-1 x h-1, 15 min before E. coli injection), and 4) posttreatment (U-78518F 30 min after E. coli injection). We measured wet-to-dry weight ratio (W/D) as an index of pulmonary edema and concentration ratios of 125I-labeled albumin in lung tissue and bronchoalveolar lavage fluid compared with plasma (L/P and BAL/P, respectively) as indexes of lung protein fluxes. In septic guinea pigs, pretreatment with U-78518F attenuated W/D, L/P, and BAL/P and posttreatment attenuated W/D and BAL/P (P < 0.05 for each). Furthermore, we studied the effect of U-78518F on human neutrophil oxygen radical production (ORP) by using flow cytometry to assess intracellular ORP and lucigenin-dependent chemiluminescence to assess extracellular ORP. Neutrophils (5 x 10(5) were stimulated with 0.5 micrograms/ml of phorbol myristate acetate. With flow cytometry, we measured intracellular ORP, cross-sectional cell area, and degranulation in neutrophils. U-78518F (minimum concn 1.0 microM) decreased intracellular ORP (n = 4; P < 0.05) when the dihydrorhodamine 123 assay was used. U-78518F (minimum concn 1.0 microM) inhibited phorbol myristate acetate-induced neutrophil chemiluminescence (n = 4; P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of chronic renal failure on skeletal and diaphragmatic muscle contraction.

The changes in muscle mechanical properties caused by the myopathy of chronic uremia was examined in the soleus and the diaphragm muscles of rats in which chronic uremia was produced by subtotal nephrectomy. Using an in vitro muscle preparation in two groups of rats (moderately and severely uremic), we determined that uremia had a significant detrimental effect on both muscles with respect to the force-frequency relationships. In the diaphragm it decreased by 15% in the moderate group and by 43% in the severe group. In the soleus it decreased by 20% in both groups. Twitch characteristics behaved differently in the soleus and the diaphragm muscles in that 1/2 RT and TPT increased significantly (p < 0.05) in the soleus (severely uremic group) but not in the diaphragm. Fatigability was increased in both muscles in the moderately uremic rats and in the diaphragm in the severely uremic rats; however, the fatigability of the soleus in the severely uremic group was not different from that in the control group. Our findings suggest that the myopathic changes occurring in chronic uremia affects the function of the soleus and the diaphragm in different ways. Other findings in the severely uremic group indicate that additional factors such as marked electrolyte imbalances may also affect the excitation-contraction coupling in different ways.

Effects of hyperphosphatemia on diaphragmatic strength and endurance.

Effects of an infusion of Na2HPO4 on diaphragm strength, endurance, and magnitude of recovery were evaluated in in situ canine diaphragm strips. Results showed no effect on maximal isometric tetanic tension. Twitch tension and tension in the low- (10-Hz) frequency range were significantly increased (P less than 0.01). Time to fatigue (endurance) increased by 38 +/- 4.5% in the group that received phosphorus compared with its control and decreased by 18.5 +/- 2.5% in the group that received dextrose compared with its control (P less than 0.005). Recovery from fatigue was also significantly improved after the phosphorus infusion. Serum ATP and 2,3-diphosphoglycerate levels were unchanged throughout the experiment. The results of this study support the notion that hyperphosphatemia improves diaphragmatic endurance and recovery from fatigue. The mechanisms involved may in part be due to the phosphate-buffering effects, which limit the extent of the muscle intracellular acidosis produced with fatigue.

Effect of dietary sodium on the severity of bronchial asthma.

BACKGROUND: A high sodium intake has been found to increase bronchial reactivity in men with asthma. The effects of change in sodium intake on peak flow rate have not been determined. METHODS: The effect of changing dietary salt intake for two weeks on the severity of asthma as measured by peak expiratory flow (PEF) was studied in 17 patients with mild asthma in an open randomised crossover trial. PEF measurements were made by the patients in their own homes. Patients were placed on three levels of dietary sodium intake: normal, low, and high. Sodium intake was assessed by 24 hour urine collection. RESULTS: The mean (SD) urine sodium was 147 (45), 84 (32), and 201 (73) mmol/24 hours in the normal, low, and high sodium intake periods respectively. There were no significant differences in PEF or PEF amplitude (highest--lowest PEF), an index of asthma lability, between the three dietary salt periods. CONCLUSION A low and high dietary salt intake for two weeks has no effect on peak expiratory flow in patients with mild asthma.