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In a previous report we found Chlamydia (C.) pneumoniae to be present in middle ear fluids (MEE) from older children with otitis media with effusion (OME). However, OME is a disease of younger children the present study was conducted in toddlers. MEEs (n = 150) and nasopharynx samples (n = 83) from children with OME was analyzed by PCR for the presence of C. pneumoniae and Mycoplasma (M.) pneumoniae. C. pneumoniae was not present in any and M. pneumoniae only in one of the MEEs, and these pathogens could be demonstrated in one and two nasopharynx samples, respectively. Further, 133 MEEs were analyzed by PCR for the presence of the 8 human herpesviruses, and all samples were found to be negative. We conclude that neither C. pneumoniae or M. pneumoniae, or any of the 8 human herpesviruses seems to play a major role in the pathogenesis of OME in early childhood OME.

The performance of an expression system based on a fusion of the bacteriophage 434-repressor to the VP16 activation domain of Herpes simplex virus type 1 (434:VP16) was tested after stable integration into Arabidopsis. A special feature of this system was the use of the monocot maize ubiquitin1 and rice actin1 promoters to drive the expression of the 434:VP16 activator and 434-repressor, respectively. Our results demonstrate that the maize ubiquitin1 and the rice actin1 promoters, each of which contain introns, are active in Arabidopsis and can be used to express genes in this dicot species. Activation of gene expression after co-integration of the activator and reporter cassettes into the same genomic locus resulted in a higher activation level (84-fold activation) compared to crossing individual lines expressing only the activator or the operator reporter cassette alone (9-fold activation). Increasing the number of operator elements in the reporter cassette from 1 to 4 increased the activation level in cross-activated lines to an average of 281-fold with one combination of parental lines giving a 900-fold activation. Simultaneous expression of the 434-repressor protein driven by the rice actin promoter resulted in a significant decrease in the 434:VP16 mediated reporter gene expression. Nevertheless, an overall induction via 434:VP16 was possible even in the presence of the 434-repressor protein. This feature is important for genes which need to be absolutely repressed except under activating conditions. To our knowledge this investigation is the first report on the use of the 434:VP16 chimeric activator in an expression system in stably transformed plant lines.

The role of high-fat diets in weight gain and obesity has been questioned because of inconsistent reports in the literature concerning the efficacy of ad libitum low-fat diets to reduce body weight. We conducted a meta-analysis of weight loss occurring on ad libitum low-fat diets in intervention trials, and analysed the relationship between initial body weight and weight loss. We selected controlled trials lasting more than 2 months comparing ad libitum low-fat diets with a control group consuming their habitual diet or a medium-fat diet ad libitum published from 1966 to 1998. Data were included from 16 trials with a duration of 2-12 months, involving 1728 individuals. No trials on obese subjects fulfilled the inclusion criteria. The weighted difference in weight loss between intervention and control groups was 2.55 kg (95% CI, 1.5-3.5; P < 0.0001). Weight loss was positively and independently related to pre-treatment body weight (r = 0.52, P < 0.05) and to reduction in the percentage of energy as fat (0.37 kg/%, P < 0.005) in unweighted analysis. Extrapolated to a BMI of about 30 kg/m2 and assuming a 10% reduction in dietary fat, the predicted weight loss would be 4.4 kg (95% CI, 2.0 to -6.8 kg). Because weight loss was not the primary aim in 12 of the 16 studies, it is unlikely that voluntary energy restriction contributed to the weight loss. Although there is no evidence that a high intake of simple sugars contributes to passive overconsumption, carbohydrate foods with a low glycaemic index may be more satiating and exert more beneficial effects on insulin resistance and cardiovascular risk factors. Moreover, an increase in protein content up to 25% of total energy may also contribute to reducing total energy intake. In conclusion, a low-fat diet, high in protein and fibre-rich carbohydrates, mainly from different vegetables, fruits and whole grains, is highly satiating for fewer calories than fatty foods. This diet composition provides good sources of vitamins, minerals, trace elements and fibre, and may have the most beneficial effect on blood lipids and blood-pressure levels. A reduction in dietary fat without restriction of total energy intake prevents weight gain in subjects of normal weight and produces a weight loss in overweight subjects, which is highly relevant for public health.

Crush syndrome or traumatic rhabdomyolysis constitutes the systemic changes seen after crush injury, i.e. the damages seen after a prolonged period of pressure on a muscle group. The pressure causes necrosis of muscle, and during revascularisation diffusion of calcium, sodium and water into the damaged muscle cells is seen, together with loss of potassium, phosphate, lactic acid, myoglobin and creatinine kinase. Untreated these changes can lead to: hyperkalaemia, acidosis, acute renal failure and hypovolaemic shock. Treatment of the systemic changes should be initiated immediately, aiming at a rapid correction of the extracellular volume and forced mannitol-alkaline diuresis. If renal failure develops, haemodialysis is started. The crush injuries are treated conservatively without fasciotomy, despite high or increasing intracompartmental pressure. The only indications for fasciotomy are lack of a distal pulse or open lesions. If fasciotomy is performed, radical removal of all necrotic muscle is essential.

A 43-year old man with alcohol abuse had septicaemia with Salmonella enteritidis. After osteosynthesis of an intertrochanteric femoral fracture he developed a deep wound infection with Salmonella enteritidis. The patient was treated with surgical debridement and antibiotics. As a consequence of alcohol abuse the patient may have had a predisposing immunosuppression, and infection with Salmonella enteritidis should be considered in case of postoperative infection in these patients.

In this study the polymerase chain reaction was used to test for the presence of Chlamydia pneumoniae DNA in 118 middle-ear aspirates from 20 children with acute otitis media (AOM) and 53 children with otitis media with effusion (OME). C. pneumoniae was detected in 8 samples obtained from 5 children with OME and, together with Streptococcus pneumoniae, in a sample from 1 child with AOM. The mean age of these five children (6.6 +/- 1.4 years) was significantly higher than that of the 48 children with OME in whom C. pneumoniae could not be detected (4.3 +/- 1.9 years). The presence of C. pneumoniae in 9.4% of the examined children with OME suggests that C. pneumoniae might be a significant supplementary factor in the etiology of this common children's disease.

A modified method for measurement of intracellular killing of Staphylococcus aureus in human neutrophil granulocytes is described. After phagocytosis of S. aureus the extracellular bacteria were sufficiently removed by repeated centrifugations and washings of the granulocytes. The optimal conditions for incubation of granulocytes for measurement of intracellular killing were found to be 37 degrees C in the presence of 5% CO2. Under these conditions, stable pH, the viability and the capacity of the granulocytes for superoxide anion generation were preserved. The number of intracellular viable bacteria was determined after lysis of the granulocytes, which should be done in H2O at pH 11 to ensure sufficient cell lysis. The kinetics and individual variation of the intracellular killing are described. The intra- or extracellular location of surviving bacteria was studied. After approximately 8 h incubation we observed intracellular growth of S. aureus followed by lysis of granulocytes and extracellular growth of bacteria. Consequently, the incubation period should not be extended beyond 5 to 8 h when the assay is used to study the effects of antibiotics on intracellular killing.

A stereological method has been used to estimate the number of migrated polymorphonuclear leukocytes in a multipore filter assay. The estimates have been compared to those obtained with conventional 51Cr-labelling methods. The precision of the estimates in individual subjects with the two methods was similar, and in both cases the contribution to the interindividual variance made by the methods was small compared to the biological variance. However, the 51Cr-labelling method resulted in a significant reduction in the fraction of neutrophils that migrated as well as a delay in the chemotactic response. Optimal assay conditions with regard to sample size, cell concentration, incubation time and chemoattractant fMLP concentration have been established for the stereological method and the virtues of the two techniques discussed.

Chemotaxis of circulating peripheral neutrophils was studied in 12 patients undergoing elective cardiac surgery. Tests of neutrophil chemotaxis were made preoperatively and 16-20 hours postoperatively, using a multipore filter assay. Statistically significant postoperative decrease was demonstrated in both stimulated and random neutrophil migration (p = 0.012 and p = 0.009, respectively). Although the peripheral neutrophil count showed a statistically significant postoperative increase (p = 0.001), the increase did not correlate to the decrease in neutrophil chemotaxis. Nor could correlation be demonstrated between the altered neutrophil chemotaxis and any per- or postoperative variables.

The acute phase C-reactive protein (CRP) was measured in serum of HIV-infected patients suffering from Pneumocystis carinii pneumonia (PCP) (32 patients), bacterial pneumonia (10 patients), and in 19 immunocompetent patients with bacterial pneumonia. The HIV-infected patients with bacterial pneumonia had a significantly lower CRP level than the immunocompetent patients (50% versus 95% had an s-CRP level > 80 mg/l). No significant difference was found in the CRP response to P. carinii or bacteria in HIV-infected patients with pneumonia due to these microorganisms (20% versus 50% had s-CRP > 80 mg/l). In the group of PCP patients, a significantly lower CRP level was found in those with CD4 positive lymphocyte counts below 50 x 10(6)/l. There was no correlation between the CRP response and the severity of the PCP as estimated by the degree of hypoxia. We conclude that the CRP level cannot be used to discriminate between PCP and bacterial pneumonia in HIV-infected patients.

A case of meningitis in a 42-year-old previously healthy man caused by the gram-negative rod Capnocytophaga carnimorsus is presented. The patient had erythema nodosum. Bacteremia was possibly induced during dental treatment.