Seroreactivity for spotted fever rickettsiae and co-infections with other tick-borne agents among habitants in central and southern Sweden.

Patients seeking medical care with erythema migrans or flu-like symptoms after suspected or observed tick bite in the southeast of Sweden and previously investigated for Borrelia spp. and/or Anaplasma sp. were retrospectively examined for serological evidence of rickettsial infection (Study 1). Twenty of 206 patients had IgG and/or IgM antibodies to Rickettsia spp. equal to or higher than the cut-off titre of 1:64. Seven of these 20 patients showed seroconversion indicative of recent or current infection and 13 patients had titres compatible with past infection, of which five patients were judged as probable infection. Of 19 patients with medical records, 11 were positive for Borrelia spp. as well, and for Anaplasma sp., one was judged as positive. Five of the 19 patients had antibodies against all three pathogens. Erythema migrans or rash was observed at all combinations of seroreactivity, with symptoms including fever, muscle pain, headache and respiratory problems. The results were compared by screening an additional 159 patients (Study 2) primarily sampled for the analysis of Borrelia spp. or Mycoplasma pneumoniae. Sixteen of these patients were seroreactive for Rickettsia spp., of which five were judged as recent or current infection. Symptoms of arthritis, fever, cough and rash were predominant. In 80 blood donors without clinical symptoms, approximately 1 % were seroreactive for Rickettsia spp., interpreted as past infection. The study shows that both single and co-infections do occur, which illustrate the complexity in the clinical picture and a need for further studies to fully understand how these patients should best be treated.

Diurnal and intra-urban particle concentrations in relation to windspeed and stability during the dry season in three African cities.

The spatial and temporal variations of PM(2.5), PM(10) and TSP in three African cities of different sizes (Dar es Salaam, Ouagadougou and Gaborone) were investigated using portable particle counters. Three different areas (downtown, green residential and traditional residential) and a reference site were designated in each of the cities in order to detect intra-urban and temporal variability. Morning, noon and night measurements were conducted in the urban areas while observations at reference stations were made continuously over the field periods. A clear diurnal pattern in particle concentrations was found in inland Gaborone and Ouagadougou, with morning and night peaks where the latter was the highest. However, in coastal Dar es Salaam the night peak was almost absent due to delayed stabilisation of the air. Particle concentrations at the Ouagadougou reference station were extreme. The direct contribution of vehicle emissions are of secondary importance since the PM(2.5)/PM(10) ratios are low (0.1-0.3). Much of the particles are supposed to be soil particles that are entrained in the air by daytime high windspeeds followed by nighttime subsidence as the air is stabilised and windspeed decreases. However, in all three cities, resuspension are important as areas with a network of unpaved roads showed the highest concentrations of suspended particles. Generally, the central business district had the lowest concentrations of particulate matter.

Emergence and persistence of beta-lactamase-producing bacteria in the upper respiratory tract in children treated with beta-lactam antibiotics.

PURPOSE: To assess the ecologic impact, in terms of selection of beta-lactamase-producing respiratory tract bacteria, of a single course of peroral beta-lactam antibiotics. PATIENTS AND METHODS: One-hundred fifty consecutive children with clinical signs of bacterial respiratory tract infection were randomly assigned to a seven-day course of treatment with either penicillin V, amoxicillin, or cefaclor. Bacteriologic specimens were collected before treatment, at its termination, and at follow-up four weeks later. RESULTS: All three drugs investigated caused a similar increase in beta-lactamase-producing bacteria, both in absolute and relative terms, an increase that persisted over a period of at least one month after completion of treatment. CONCLUSION: Penicillin V, amoxicillin, and cefaclor all act as selective agents for beta-lactamase-producing bacteria in the upper respiratory tract. Treatment with a peroral beta-lactam antibiotic puts patients at risk of becoming persistent carriers of beta-lactamase-producing bacteria.

Serological identification of Branhamella catarrhalis. Serological evidence for infection.

A protein antigen--P-antigen--characteristic of Branhamella catarrhalis has been described. Precipitating antibodies against this antigen occur in a majority of healthy human sera; serological evidence for a pathogenic role of B. catarrhalis is accumulating. The occurrence of complement-fixing antibodies has been demonstrated during bronchopulmonary infection with B. catarrhalis and in maxillary sinusitis. Increases in antibody titres assessed by an enzyme-linked immunosorbent assay (ELISA) technique have been reported in children with acute otitis media. An ELISA, using purified P-antigen, for the evaluation of IgG, IgM and IgA antibodies against B. catarrhalis is being developed.

Upper respiratory tract infections. Ecological and therapeutic aspects of beta-lactamase production with special reference to Branhamella catarrhalis.

Available data indicate that the most common beta-lactamase produced by Branhamella catarrhalis is plasmid mediated. The same enzyme occurs in Moraxella nonliquefaciens, a commensal in the upper respiratory tract. The ability to produce the enzyme, which is known as BRO-1, can be transferred by conjugation from M. nonliquefaciens to B. catarrhalis. Since the first beta-lactamase-producing strains of B. catarrhalis appeared in 1977, the frequency of beta-lactamase production has increased rapidly; figures as high as 76% have been reported. The plasmid-mediated beta-lactamase TEM-1 occurs in several species of the genus Haemophilus. While the frequency of beta-lactamase production in H. influenzae is reported to be 10-15%, the incidence is significantly higher in non-pathogenic Haemophilus species. Both phenoxymethyl-penicillin and ampicillin promote the occurrence of beta-lactamase-producing strains, but the selective pressure exerted by ampicillin seems to be more pronounced. It may be possible to reduce the ecological effects of the penicillins by avoiding overdiagnosis of the most common bacterial infections of the respiratory tract, and by shortening the courses of antibiotic treatment.

Plasmid-mediated beta-lactamase in Branhamella catarrhalis.

The plasmid-mediated beta-lactamase in Branhamella catarrhalis (BRO-1), also occurring in Moraxella nonliquefaciens, differs from other known plasmid-mediated beta-lactamases in Gram-negative bacteria regarding substrate profile and isoelectric point. B. catarrhalis strains previously reported to produce beta-lactamases deviating from BRO-1 were tested, and the beta-lactamases did not differ significantly from BRO-1 in substrate profile, isoelectric point or relative substrate affinity index (RSAI). Further investigations of strains of various geographic origin should be undertaken. RSAI seems to be a useful tool for screening of beta-lactamases in B. catarrhalis since values for a large number of strains can easily be determined. The previously reported conjugational transfer of BRO-1 production within species B. catarrhalis and from M. nonliquefaciens to B. catarrhalis was confirmed. Four bands of extrachromosomal DNA were regularly detected by agarose gel electrophoresis in beta-lactamase-producing as well as in beta-lactamase-negative strains of B. catarrhalis and M. non-liquefaciens, provided that the excessive nuclease activity in the preparations was inhibited.

Possible role of short-term parenteral nutrition with fat emulsions for development of haemophagocytosis with multiple organ failure in a patient with traumatic brain injury.

We describe a case of life-threatening haemophagocytosis after a short term nutrition with fat emulsion in a 21-year-old woman who sustained an isolated traumatic brain injury. Hypertriglyceridemia and "creaming plasma" were observed after a three-day period of parenteral fat nutrition (Intralipid 20%). She also developed rash, high fever (40-41 degrees C), hypertension, raised intracranial pressure, hepatic and renal failure, haemolysis, marked thrombocyto- and leucopenia, coagulation disorder and pulmonary failure. These symptoms, together with a typical bone marrow smear, indicated haemophagocytosis with hyperactivation of the monocyte-macrophage system. We suggest that the hyperactivation was an effect of fat retention or agglutination of the fat particles; the initial triggering mechanism may emanate from the brain damage by hypercytokinaemia. The steroid treatment given most likely contributed to the successful outcome, as indicated by the stepwise improvement related in time to the steroid infusions.

Expression of the myc protooncogene in canine rhabdomyosarcoma.

The expression of the protooncogene c-myc in a canine rhabdomyosarcoma was examined. It was found that this highly malignant tumour contained vast quantities of RNA that hybridized with a cDNA probe for c-myc. Restriction fragment length analysis after endonuclease digestion of tumour DNA did not reveal any rearrangements in this gene locus. The potential role of this oncogene in the development of canine rhabdomyosarcoma is discussed.

Poly-L-arginine and an N-formylated chemotactic peptide act synergistically with lectins and calcium ionophore to induce intense chemiluminescence and superoxide production in human blood leukocytes. Modulation by metabolic inhibitors, sugars, and polyelectrolytes.

Various cationic polyelectrolytes (poly-alpha-amino acids and histones), lectins, the chemotactic peptide, f-methionyl-leucyl-phenylalanine (fMLP), the calcium ionophore A23187, and phorbol myristate acetate (PMA) were investigated regarding their capacity to induce luminol-dependent chemiluminescence (LDCL) and superoxide production by human blood leukocytes. Although when tested individually, poly-L-arginine (PARG), phytohemagglutinin (PHA), concanavalin A (Con A), or fMLP induced only a low to moderate LDCL response, very intense synergistic CL reactions were obtained by mixtures of PARG + PHA, PARG + Con A, PARG + PHA + fMLP, Ca2 + ionophore + PARG + PHA + fMLP, and PARG + PMA. The sequence of addition of the various agents to WBC in the presence of luminol absolutely determined the intensity of the LDCL signals obtained, the highest reactions being achieved when the WBC were preincubated for 2-3 min with A23187 followed by the sequential addition of fMLP, PARG, and PHA. These "multiple hits" induced CL reactions which were many times higher than those obtained by each factor alone. On the other hand, neither poly-L-lysine, poly-L-ornithine, poly-L-histidine, nor poly-L-asparagine, when employed at equimolar concentrations, cooperated efficiently with PHA and fMLP to trigger synergistic LDCL responses in leukocytes. Concomitantly with the induction of LDCL, certain ligand mixtures also triggered the production of superoxide. The LDCL which was induced by the "cocktail" of agents was markedly inhibited by sodium azide (93% inhibition), but to a lesser extent by catalase (10% inhibition) or by superoxide dismutase (20%-60% inhibition). On the other hand, scavengers of singlet oxygen and OH (sodium benzoate, histidine) did not affect the synergistic LDCL responses induced by these multiple ligands. Cytochalasin B also markedly inhibited the LDCL responses induced either by soluble stimuli or by streptococci preopsonized either with histone or with polyanethole sulfonate. The LDCL responses which were induced by mixtures of PARG and concanavalin A were also strongly inhibited by mannose, alpha-methyl mannoside, and poly-L-glutamic acid. The data suggest that the LDCL responses induced by the soluble ligands involved a myeloperoxidase-catalyzed reaction. The possible employment of "cocktails" of ligands to enhance the bactericidal effects of PMNs, macrophages, and natural killer cells on microbial cells and mammalian targets is discussed.

Erythromycin in acute laryngitis in adults.

Moraxella catarrhalis and Hemophilus influenzae are isolated from the nasopharynx in 50% to 55% and 8% to 15%, respectively, of cases of acute laryngitis in adults. This finding indicates that these organisms, M catarrhalis in particular, are in some way involved in the pathogenesis of the disorder. In the present double-blind, placebo-controlled trial, the effect of erythromycin ethylsuccinate (0.5 g twice a day for 5 days) on the elimination of nasopharyngeal pathogens and reduction of clinical signs of upper respiratory tract infection, as well as on subjective complaints, was evaluated in 106 adults with acute laryngitis. The bacterial isolation rates at presentation were M catarrhalis 50%, H influenzae 18%, and Streptococcus pneumoniae 4%. In the 99 patients who completed the study, the elimination of M catarrhalis after 1 week was better in the erythromycin group (25 of 30 cases) than in the placebo group (6 of 19 cases; p < or = .00038). The elimination of H influenzae was unaffected by erythromycin. Otolaryngologic examination did not reveal any significant group differences regarding laryngitis, pharyngitis, or rhinitis. Voice quality was improved after 1 week, irrespective of treatment. However, as compared to the placebo group, the erythromycin group reported fewer voice complaints after 1 week and fewer coughing complaints after 2 weeks. As acute laryngitis in adults is self-limiting, and subjective symptoms are spontaneously reduced after 1 week in most cases, antibiotic treatment does not seem warranted as a general policy. However, erythromycin may be justified in patients who are professionally dependent on voice function.

Inefficacy of penicillin V in acute laryngitis in adults. Evaluation from results of double-blind study.

Patients with acute laryngitis following an upper respiratory tract infection are often treated with antibiotics for their voice complaints, although, to our knowledge, the effect of such therapy has not been examined. In the present study, comprising 100 adults with laryngitis, the rate of resolution of vocal symptoms, as estimated from voice recordings or subjectively by the patients, was the same in patients who received penicillin V (pcV) as in those who received placebo. Similarly, the degree of rhinorrhea/nasal congestion and cough was not significantly influenced by pcV treatment. At the acute visit, nasopharyngeal cultures revealed Branhamella catarrhalis in 50%, Hemophilus influenzae in 15% and Streptococcus pneumoniae in 1% of the patients; the rate of elimination of these bacteria was the same in the pcV as in the placebo group. Thus, while suggesting that B catarrhalis and H influenzae are important for the pathogenesis of the disorder, our results do not provide support for the use of pcV in acute laryngitis.

Thermal oral-pharyngeal stimulation and elicitation of swallowing.

The influence of thermal stimulation of the oral and pharyngeal mucosa on the elicitation of swallowing was studied in 14 healthy volunteers. The Repeated Dry Swallowing Test was used to study the ability to elicit swallows. The test consists of 11 manometrically recorded dry swallows performed at maximum speed. The time between swallows 1 and 11 (Swallowing Test Time) is considered a measure of the ease of eliciting the swallowing reflex. The study includes two controlled experiments, each consisting of an active test done immediately after a cold stimulus and a control done after a stimulus at body temperature. In one experiment the stimulus consisted of stimulation of the fauces with a laryngeal mirror and in the other of swallowing water. Each subject served as his or her own control. The order of the tests was randomized and the results read blindly. In the mirror experiment, the swallowing test times obtained after application of a cold laryngeal mirror did not differ significantly from those obtained after stimulation at body temperature. In the water experiment, swallowing test times were shorter after swallowing cold water compared to those after swallowing water at body temperature in 11 of the 14 volunteers (n.s.). The concept that swallowing is significantly facilitated by cold applied to the oral-pharyngeal mucosa was thus not supported by the present study. The importance of these findings is discussed.

[Does the infectious disease, human ehrlichiosis, exist in Sweden? Ticks get a hold of new zoonoses]. / Finns infektionssjukdomen human ehrlichios i Sverige? Fästingar fäste för nya zoonoser.

Human ehrlichiosis diseases, decently recognised as emerging human infections in the USA, are caused by vector-borne, strictly intracellular bacteria of the family Rickettsiaceae. Human monocytic ehrlichiosis is caused by Ehrlichia schaffeensis, whereas the agent causing human granulocytic ehrlichiosis (HGE) has yet to be identified [1]. The putative increase in the occurrence of these primary zoonoses is dependent on the complex relationship between the infectious agents, the vectors, and the hosts (rodents, deer) which constitute the wild-life reservoir. In Scandinavia, granulocytic ehrlichiosis is well known in veterinary medicine. Pasture fever in cattle and sheep is caused by Ehrlichia phagocytophila, whereas granulocytic ehrlichiosis in horses and dogs is caused by a new, recently characterised Ehrlichia species [2]. All species of Ehrlichia causing granulocytic ehrlichiosis are closely related both genetically and antigenically, and are all transmitted by ticks of the genus Ixodes. In Sweden, veterinary cases of granulocytic ehrlichiosis are characterised by a geographical distribution corresponding well with that of Ixodes ricinus, and a seasonal distribution similar to that of Lyme borreliosis. In Europe, clinical cases of HGE have so far been reported only from Slovenia [5], though seroprevalence figures of 8-17 per cent have been reported for tick-exposed populations [6, 7]. As most cases are probably subclinical, and as clinical symptoms, when present, are non-specific, clinical diagnosis is dependent on the clinician's awareness of the existence of the disease. Laboratory diagnostic tests are now available at Kalmar.

[Varying clinical picture and course of human granulocytic ehrlichiosis. Twelve Scandinavian cases of the new tick-borne zoonosis are presented]. / Varierande klinisk bild och förlopp vid human granulocytär ehrlichios. Tolv skandinaviska fall av den nya fästingburna zoonosen redovisas.

In the twelve clinical cases of human granulocytic ehrlichiosis (HGE) so far identified in Scandinavia (ten in Sweden, two in Norway), clinical presentation varied from a mild febrile illness to a severe septic condition with such systemic complications as acute respiratory distress syndrome (ARDS). Laboratory verification was based on PCR (polymerase chain reaction) in ten cases, and on serology in two cases. Sequence analysis of 16S rDNA showed the infectious agents to belong to the Ehrlichia phagocytophila genogroup. Seroprevalence data indicate widespread human exposure to granulocytic Ehrlichia; mean seroprevalence, 15-20% of 1,000 clinical sera from tick-exposed patients (mainly from Sweden and Norway). Proposals for diagnostic criteria and procedures, and case management are presented in the article.

Intra-urban air pollution in a rapidly growing Sahelian city.

In this paper we analyze spatial and temporal variations of air pollution (PM(1), PM(2.5), PM(10), CO, NO(x), O(3), Toluene and Benzene) and climate in areas of different development typology in Ouagadougou, Burkina Faso. Analyses are based on measurements from fixed sites and car traverse measurements during field studies in 2007 and 2010. Large spatial and temporal variations were found, showing a generally poor air quality situation, with extreme levels of PM(10), commonly exceeding air quality guidelines of WHO. Pollution levels increase considerably with increased atmospheric stability. Important sources were transported dust and re-suspension of dust from unpaved roads, but also traffic emissions and biomass burning. The spatial variations are examined with focus on effects for variations in potential exposure depending on for example area of residence and daily activity pattern, showing that great differences are likely to exist. Ouagadougou, like most developing countries worldwide, currently experiences an extremely rapid population growth in combination with limited financial means. This is likely to create increasingly harmful air pollution situations for the rapidly growing populations of these areas, and shows an urgent need for increased understanding of the pollution situation as well as development of mitigation strategies.

C6 peptide ELISA test in the serodiagnosis of Lyme borreliosis in Sweden.

The aim of this study was to evaluate the synthetic C6 peptide test as a first-line test in a two-tiered scheme for Borrelia serology in a clinically well-characterized population of patients with Lyme borreliosis in Kalmar County, Sweden. The study population consisted of a prospective group (n = 200), a control group (n = 255), and a retrospective group (n = 29). The test panel consisted of the Immunetics Quick ELISA C6 Borrelia assay kit (Immunetics, Cambridge, MA, USA), the Virotech Borrelia burgdorferi ELISA (Genzyme Virotech, Rüsselsheim, Germany), and the Liaison Borrelia CLIA (DiaSorin, Saluggia, Vercelli, Italy). Seroprevalence among 200 healthy blood donors was significantly lower in the C6 test (8%) compared to the Virotech ELISA (14%) and the Liaison CLIA (12%). In convalescent sera (2-3 months and 6 months post infection) from 158 patients with erythema migrans, the seropositivity in the C6 test was also significantly lower compared to both the Virotech ELISA and the Liaison CLIA. Serosensitivity in the acute phase of erythema migrans and other clinical manifestations of borreliosis did not differ significantly between the C6 test and the Virotech ELISA or the Liaison CLIA. Overall, a positive C6 test seems to correlate well with acute borreliosis. Cross-reactivity was lower in the C6 test in sera positive for Epstein-Barr virus infection as compared to the Virotech ELISA. This study supports the use of the C6 test as a screening test for borreliosis, in endemic areas.

Reduced number of interleukin-12 secreting cells in patients with Lyme borreliosis previously exposed to Anaplasma phagocytophilum.

Lyme borreliosis and human granulocytic ehrlichiosis are tick-borne diseases caused by Borrelia burgdorferi and Anaplasma phagocytophilum, respectively. Infection with A. phagocytophilum has been observed to induce immunosuppression and animal studies suggest that the bacteria might also have prolonged inhibitory effects on immune cells. The aim of this study was to investigate the cytokine secretion in patients exposed previously to A. phagocytophilum and currently infected with B. burgdorferi compared with patients infected with B. burgdorferi and seronegative for A. phagocytophilum. Eight patients with erythema migrans and antibodies against A. phagocytophilum, 15 patients with erythema migrans and negative A. phagocytophilum serology and 15 non-exposed healthy individuals were included in the study. Blood mononuclear cells were stimulated with Borrelia-antigen and the number of cytokine [interleukin (IL)-4, IL-5, IL-12, IL-13 and interferon (IFN)-gamma]-secreting cells was detected by enzyme-linked immunospot (ELISPOT). This study shows that patients with a previous exposure to A. phagocytophilum and a current infection with B. burgdorferi have a lower number of Borrelia-specific cells secreting IL-12 compared to Ap seronegative patients infected with B. burgdorferi (P < 0.001), indicating impairment in the ability to mount strong Th1-responses. We suggest that this mirrors a reduced Th1 response caused by A. phagocytophilum which could influence the outcome of the Borrelia infection and, speculatively, may also have implications in other conditions.

A protein antigen characteristic of Branhamella catarrhalis. Serological identification of the genus.

Precipitation patterns of sonicated, acid-extracted and other extracts from Branhamella catarrhalis were examined by double diffusion-in-gel technique, using antiserum to B. catarrhalis. Acid extract gave rise to 4 distinct precipitates. One of these lines was further studied. The bacterial component responsible for this line was trypsin-sensitive, indicating that it was a protein. It was anodally localized by crossed immunoelectrophoresis. By absorption of antiserum with whole bacteria, the precipitating capacity of the serum was diminished, suggesting that the protein antigen (P-antigen) was exposed on the bacterial surface. F(ab')22-fragments of IgG from antiserum, but not from normal rabbit serum, precipitated the P-antigen, indicating that it was a true antigen-antibody reaction. It was possible to make an IgG preparation monospecific for the P-antigen, by absorbing antiserum with trypsinized bacterial extract. 31 strains of B. catarrhalis, 9 strains of N. gonorrhoeae, 10 strains of N. meningitis, 12 other Neisseria spp. and 2 strains of H. influenzae were investigated for presence of cros-reacting surface antigens, using IgG monospecific for the P-antigen and 125I-labelled protein A from Staphylococcus aureus. After antibody exposure, all 31 strains of B. catarrhalis showed abundant uptake of protein A. No significant uptake was detected on any other investigated strain. Hence, the P-antigen appears to be characteristic of B. catarrhalis. The possibility of a serological identification of the species is introduced. Precipitating antibodies against the P-antigen were demonstrated in 69% of normal human sera.

Characterization of the plasmid-mediated beta-lactamase in Branhamella catarrhalis, with special reference to substrate affinity.

The plasmid-mediated Branhamella catarrhalis beta-lactamase BRO-1, also found in Moraxella nonliquefaciens, was characterized as regards substrate profile, isoelectric point and relative substrate affinity index (RSAI) to various substrates and compared in these aspects with the TEM-1 enzyme of Haemophilus influenzae. As measured by a biological assay and with high performance liquid chromatography (HPLC), BRO-1 was found to hydrolyse carbenicillin, mecillinam, methicillin and cefaclor with a higher rate than TEM-1. The only substrates having a relative rate of hydrolysis higher for TEM-1 than for BRO-1 were ampicillin and cephaloridine. The rates of hydrolysis registered with these two methods were comparable for all but 2 of 13 tested substrates. Isoelectric focusing yielded a main band at pH 5.6 and several satellite bands consistent with those reported by other authors for Branhamella enzymes having a substrate profile similar to that of BRO-1. A tenfold or higher difference in RSAI between BRO-1 and TEM-1 was recorded for five of the 15 compounds tested. BRO-1 seems to be the most common beta-lactamase in Bran. catarrhalis, irrespective of geographic origin. Its substrate profile, isoelectric pattern and RSAI differ from those of other known plasmid-mediated beta-lactamases described, thus justifying a specific designation.

Beta-lactamase production in the upper respiratory tract flora.

In order to determine the recovery rate of species of the genera Haemophilus and Moraxella (including subgenus Branhamella) from the upper respiratory tract and the incidence of beta-lactamase production within these genera, cultures were made of nose and throat swab specimens and adenoid tissue in 50 children undergoing adenoidectomy. Haemophilus influenzae was isolated from 92% of the children. All children harboured strains of Haemophilus spp. and in 46%, at least one strain produced the TEM-1 beta-lactamase. Branhamella catarrhalis and/or Moraxella nonliquefaciens were isolated from 82% of the children and strains producing the BRO-1 beta-lactamase from 34%. Overall, TEM-1 and/or BRO-1 producing strains were recovered from 60% of the investigated patients. The beta-lactamase production was found to be transferable by conjugation within the respective genera. It is suggested that the apathogenic species may be a source of transferable determinants mediating beta-lactamase production in the upper respiratory tract.