Validation of a multiplex reverse transcriptase PCR ELISA for the detection of 19 respiratory tract pathogens.

INTRODUCTION: Since acute respiratory tract infections inflict a high burden of disease in children worldwide, a multiplex reverse transcription polymerase chain reaction combined with a microwell hybridization assay (m-RT-PCR-ELISA) to detect 19 different respiratory pathogens was developed and validated. METHODS: A total of 430 respiratory specimens were retrospectively tested in parallel by both the advanced 19-valent m-RT-PCR-ELISA as well as by culture or individual RT-PCR assays used in clinical routine. RESULTS: The mean (median) sensitivity of the m-RT-PCR-ELISA in the retrospective test was 93.3% (95.1%; range 83.3-100 %), and the mean (median) specificity was 99.8 and 100 % (range 98.6-100 %), respectively. The mean positive predictive value was 99.3 % (range 93.4-100 %) and the mean negative predictive value was 95.3 % (range 98.4-100 %). Feasibility and clinical value of the 19-valent method was prospectively shown on 16,231 incoming clinical specimens from patients between 0 and 16 years of age with acute respiratory tract infections admitted to pediatric hospitals or private practices from October 2003 to June 2010 in three regions in Germany (Kiel, Mainz, Freiburg; Freiburg to June 2007 only). At least one microorganism was detected in 10,765 of 16,231 (66.3 %) clinical specimens: 5,044 RV, 1,999 RSV, 1,286 AV, 944 EV, 737 seasonal IVA, 173 pandemic IVA H1N1-2009, 899 MPV, 518 CV, 383 PIV3, 268 PIV1, 259 Mpn, 205 IVB, 164 PIV2, 144 PIV4, 103 Bp, 29 Cpn and 29 Bpp, while reovirus and Lpn were not present in these specimens from a pediatric population. More than one organism could be detected in 13.4 % of the specimens. CONCLUSIONS: The m-RT-PCR-ELISA evaluated here improves the spectrum for diagnosing respiratory infections and is a feasible instrument for individual diagnostic and epidemiological studies.

Comparison of the BD Directigen Flu A+B Kit and the Abbott TestPack RSV with a multiplex RT-PCR ELISA for rapid detection of influenza viruses and respiratory syncytial virus.

The Directigen Flu A+B enzyme immunoassay and the Abbott TestPack RSV enzyme immunoassay were each compared with a multiplex RT-PCR ELISA by testing 635 nasopharyngeal aspirates collected from children aged < 16 years who had been hospitalised with acute respiratory tract infection during the epidemic season 2002-2003. In this study, the sensitivity of the Directigen Flu A+B assay was unacceptably low (29.3% and 10.0%, respectively) for the detection of influenza A and B viruses. The sensitivity of the Abbott TestPack RSV assay (77.4%) was acceptable and in agreement with the multiplex RT-PCR ELISA.

Evaluation of a multiplex reverse transcriptase PCR ELISA for the detection of nine respiratory tract pathogens.

BACKGROUND: A multiplex reverse transcription (RT) polymerase chain reaction combined with a microwell hybridization assay (m-RT-PCR-ELISA) was previously developed to detect nine different microorganisms: enterovirus (EV), influenza virus type A (IVA) and type B (IVB), respiratory syncytial virus (RSV), parainfluenzavirus type 1 (PIV1) and type 3 (PIV3), adenovirus (AV), Mycoplasma pneumoniae (Mpn), Chlamydia pneumoniae (Cpn) in a single test. These organisms do not usually colonize the respiratory tract of humans, but, if present, it may be assumed they are involved in respiratory disease. OBJECTIVES AND STUDY DESIGN: The m-RT-PCR-ELISA was tested on (i) culture supernatants of unknown contents, (ii) by determining the analytical sensitivity of 10-fold serial dilutions of culture supernatants and (iii) by determining clinical sensitivity in a retrospective study on 411 clinical specimens. The specimens were re-tested in parallel by m-RT-PCR-ELISA versus the gold standard culture and immunfluorescence, and versus individual RT-PCR. RESULTS: (i) The 9-valent m-RT-PCR-ELISA shows 83% to 100% concordant results on 103 culture supernatants containing different organisms. (ii) The analytical sensitivity was as follows: higher sensitivity of the 9-valent m-RT-PCR-ELISA in comparison to culture in the cases of PIV3, IVA and IVB (factor 10) and AV and EV (factor 100), and lower sensitivity in case of RSV and PIV1 (factor 10). (iii) The agreement with the gold standard in the kappa statistic was excellent for RSV (kappa = 0.937), IVA (kappa = 0.940), very good for PIV1 (kappa = 0.914), IVB (kappa = 0.907) and satisfactory for PIV3 (kappa = 0.410). For AV, EV and Mpn the m-RT-PCR-ELISA preliminary could be qualified as very good, based on the data derived on culture supernatants. Information about the validity for Cpn is limited. CONCLUSION: The m-RT-PCR-ELISA is a feasible, sensitive and specific method for detection of a broad spectrum of organisms. It is suitable for individual as well as epidemiological diagnosis.

Candida arthritis in a premature infant treated successfully with oral fluconazole for six months.

INTRODUCTION: Candida arthritis in premature infants is a rare condition mainly treated with parenteral amphotericin B. CLINICAL PICTURE: Arthritis 9 weeks after birth. The infection resulted out of colonisation at birth and fungaemia at day 6 and day 13 of life. TREATMENT: Oral fluconazole for 6 months (5 mg/kg/day after loading with 7 mg/kg/day). OUTCOME: Full remission. CONCLUSIONS: Oral fluconazole is an alternative to parenteral amphotericin B in the treatment of premature infants, but proof of absorption and an extended treatment period according to prematurity might be warranted.

Safety and efficacy of acellular pertussis vaccines: the Mainz study and other recent studies.

Following concerns about the safety and reactogenicity profile of diphtheria, tetanus and whole cell pertussis vaccines (DTwP), new and less reactogenic alternatives were developed over the last two decades. The new diphtheria, tetanus and acellular pertussis vaccines (DTaP) no longer consist of the whole bacterial cell but of either extracts or of a few highly purified components. While it soon became clear that DTaP vaccines are significantly less reactogenic than DTwP vaccines, their efficacy was disputed and remained unproven. First studies and epidemiological data from Japan suggested vaccine efficacy rates (VE) of about 80%; however, the first blinded clinical trial from Sweden documented a much lower VE. Worldwide, seven large DTaP efficacy trials have recently been completed. Our own efforts included a large safety trial with 22505 vaccinees and, nested in this setting, a prospective household contact study. Typical WHO-defined pertussis developed in 7 of 112 DTaP vaccinated children following household exposure as compared to 96 cases in 173 children not vaccinated against pertussis. Thus, vaccine efficacy was calculated to be 88.7% (95% CI 76.6 to 94.6). The median duration of spasmodic cough in the few children vaccinated with DTaP who did start coughing was 17 days as compared to 35 days in unvaccinated children. No waning of protection was observed. None of the confounding variables analyzed influenced study results in favour of DTaP. Following administration of more than 67000 DTaP doses, 153 serious adverse events were reported. Eight events were considered possibly related and five were considered related to the study vaccine. According to additional study results from the other trials it can be concluded that DTaP vaccines, like DTwP vaccines, are safe and effective. The choice between DTwP and DTaP should be based on acceptance of the reactogenicity profile, coverage rates achieved, costs and other factors in each individual country.

[Ultrasound hemolysis]. / Ultraschall-Hämolyse.

An ultrasound unit for hemolysis for an analytic device is presented which permits hemolysis without the need of a specific insonification chamber which is an unwanted discontinuity along the probe path. This is possible by direct insonification of a flexible tube which contains the flowing blood. The influence of ultrasound power, insonification time and coupling conditions on the result of hemolysis is investigated.

PID-ARI.net - a pediatric infectious diseases network on acute respiratory infections and the added value of a multilevel research network.

BACKGROUND: PID-ARI.net was one of three infectious disease epidemiological research networks funded by the German Ministry of Education and Research (BMBF). Its objectives were to strengthen the national initiative on infectious diseases epidemiology and to focus on a health care problem of high relevance. PATIENTS AND METHODS: A research network on the epidemiology of ARI in children was formed to generate data on several levels. Key structure was a centrally organized active surveillance system in three areas of Germany from north to south. RESULTS: In the 6 years of funding by the BMBF, an integrated research network with a known population denominator was formed. In the laboratory-based surveillance of up to 19 respiratory pathogens, 18,899 samples were analyzed. The added value is utilization of data on time, place, person and pathogen of a disease episode at several levels - from surveillance and online publication via a website to descriptive, analytical and molecular epidemiology and further specialized projects. Its wide age range including children up to 16 years of age, an extensive panel of pathogens, a known population denominator and the diversity of 3 distant geographical areas should considerably reduce vulnerability due to bias. CONCLUSIONS: Active surveillance systems for ARI are superior to passive systems. If a surveillance system such as the one used in PID-ARI.net is part of a research network which can utilize the data on several levels, the expenditure for such a system should be worthwhile and such a system would be an asset to any health care system.

Lower respiratory tract disease in children: constant pathogens - constant management?!

BACKGROUND: This study analyses the pathogens of acute lower respiratory tract infections (LRI) in children in a German community hospital over six years. Against this background the adoption of new diagnostic and therapeutic guidelines for the LRI management and of RSV-cases in particular is studied. METHODS: 1054 children aged zero to 36 months hospitalized with LRI were prospectively included in the surveillance studies "Parainfluenzavirus (PIV) and Respiratory syncytial virus (RSV) infections in Germany [PRI.de] 1999-2001" and the "pediatric infectious diseases network on acute respiratory tract infections" [PID-ARI.net] for the time period of October 2002 until June 2005. The nasopharyngeal aspirates (NPA) of these children had been analysed for RSV, PIV 1,2,3 and influenzavirus (IV)-A, -B. In 2003/2004 the national guideline on how to diagnose and treat RSV-disease (bronchiolitis) changed. Data on LRI cases severity and especially those regarding the clinical management of RSV-infections were compared to see differences following the release of the guideline. RESULTS: 84% of the children were between zero and 24 months old. 34% of the NPA specimens were positive for RSV, 7.7% for PIV 1,2,3 and 4.7% for IV-A, -B. Epidemiological findings did not differ substantially between the two studies. Clinical management of RSV-LRI, especially drug use, did not change except for the lower rate of x-ray examination (p<0.01). CONCLUSION: The spectrum of causing agents in LRI of children remained quite stable over of six years. Diagnostic and therapeutic concepts remain also stable in a situation where new guidelines were introduced, but not reinforced.

The descriptive epidemiology of severe lower respiratory tract infections in children in Kiel, Germany.

BACKGROUND: Lower respiratory tract infections (LRI) inflict a high burden of disease in children worldwide. Longitudinal, descriptive epidemiological data on different forms of LRI are urgently needed to differentiate this burden, compare population-based incidence rates between countries and to recognize trends. PATIENTS AND METHODS: From July 1996 to June 2000, all children hospitalized with LRI, i. e. laryngo-tracheo-bronchitis (LTB), bronchitis, wheezing bronchitis-bronchiolitis (WBB), bronchopneumonia and pneumonia, in the municipal area of Kiel, Schleswig-Holstein, Germany, were analyzed by cross-sectional studies. Naso-pharyngeal aspirates (NPA) were analyzed by an in-house 9-valent multiplex-RT-PCR. RESULTS: In the 4-year observational period, 1 072 children aged 0 to 16 years (median 23 months) were hospitalized with LRI: 12 % (median 28 months) with LTB, 11 % (median 17 months) with bronchitis, 28 % (median 13 months) with WBB, 26 % (median 26 months) with bronchopneumonia and 22 % (median 47 months) with pneumonia. The prevalence of chronic underlying conditions (20 %) and low gestational age (13 %) varied in the different forms of LRI. The cumulative incidence rate of LRI rose steadily over the 4 years. The highest fraction was contributed by WBB, while pneumonia declined. The highest incidence rate ratio was attributable to respiratory syncytial virus (RSV, 0.46) and to children under 2 years of age. CONCLUSIONS: LRI, especially obstructive forms of LRI, are increasing in Germany as described earlier for the USA, UK and Sweden. The major burden is carried by children under 2 years. RSV is the single pathogen with the highest impact.

Population-based incidence of severe pneumonia in children in Kiel, Germany.

BACKGROUND: Elaborated data on the descriptive epidemiology of community-acquired pneumonia (CAP) are a prerequisite to estimate the impact of new vaccines. PATIENTS AND METHODS: From July 1996 to June 2000, all children (0-16 years) admitted to one of the two pediatric hospitals in Kiel and being resident in the municipal area of Kiel were investigated by cross-sectional studies and prospective testing using a 9-valent in-house m-RT-PCR method. RESULTS: In the 4-year period, 514 children were included (mean age 46, median 40 months): 279 were diagnosed with bronchopneumonia (BPN, median age 26 months), 235 with pneumonia (PN) (47 months); within the latter 69 cases had lobar PN (55 months), 41 atypical PN (51 months) and 28 parapneumonic effusions (74 months). An underlying chronic condition was present in 22.8 % and 10.1 % were born prematurely. The population-based incidence rates (per 100,000 per year) were on average 300 for children 0-16 years, 163 for BPN, 136 for PN, 53 for lobar PN, 24 for atypical PN and 16 for parapneumonic effusions. The rate was stable or slightly declined over the observation period. 61 % of infants and 45 % of children under 5 years of age have to be hospitalized having contracted CAP. The highest fraction of 34 and 25 %, respectively, was attributable to RSV. Viruses were not diagnosed significantly more often in BPN than in PN, if stratified by age. CONCLUSION: The incidence and the admission rate of severe CAP is lower than in the USA. The high rate of empyema warrants enhanced surveillance as an indicator for antibiotic resistance or changing impact of pneumococcal serotypes. Misclassification, also with ICD codes, is a major issue. Well analyzed epidemiological recruitment areas are a valid tool to generate precise data in Germany.

RSV-prevention in children guided by a web-based early warning system.

BACKGROUND: Passive immunization with palivizumab is expensive and requires considerable logistic effort. So far 5 monthly injections from November to March are recommended. The RSV season onset and its duration, however, shows considerable variation. In many countries on the northern hemisphere a dual rhythm is described. METHOD: A web-based early warning system within the research network PID-ARI.net is in place since 2002. The surveillance data are published online weekly via www.pid-ari.net. This enables physicians to carry out interventions, like passive immunization for RSV, synchronously with the epidemiology of a given pathogen instead of a rigid schedule. The surveillance of PID-ARI.net is based on a 19 valent multiplex RT-PCR on naso-pharyngeal aspirates. The samples are provided by hospitals and offices in Freiburg, Mainz and Schleswig-Holstein (north, middle, south of Germany). Children with lower airway infections are prospectively enrolled. RESULTS: In the time period from July 1999 to June 2003 with 20 months of recommended palivizumab application, 5 months (25 %) would have been not on target. In two seasons the start of the vaccine campaign would have been too early (waste of two months). In one season the application would have started one month too late and in two seasons the vaccine campaign would have been stopped two months too early leaving the vaccinees on risk for acquiring RSV. CONCLUSIONS: The web-based early warning system of PID-ARI.net is the first, pathogen-specific, comprehensive and fast surveillance-system for airway pathogens in Europe. It facilitates the epidemic-synchronous use of the passive immunization with palivizumab and by this increases its efficiency and should safe costs.

Expression of connexins 40 and 43 in human left atrium in atrial fibrillation of different aetiologies.

OBJECTIVE: To test the hypothesis that atrial fibrillation (AF) is associated with changes in the expression of connexins 40 and 43 in the left atrium with more pronounced changes in mitral valve disease than in lone AF. METHODS: Protein concentrations of connexin 40 and connexin 43 were analysed in left atrial tissue of patients undergoing cardiac surgery. One group of patients had lone AF (n = 41), one group had AF and mitral valve repair (n = 36), and one group in sinus rhythm served as controls (n = 15). RESULTS: Western blot analysis of connexin 40 and connexin 43 expression showed an increase of both gap junctional proteins (connexin 43 > connexin 40) in patients with AF of all forms compared with patients in sinus rhythm (p = 0.01 and p = 0.011, respectively). Subgroup analysis showed increased concentrations of connexin 40 in lone AF and AF with mitral valve disease compared with sinus rhythm (p = 0.06 and p = 0.029, respectively), whereas the same analysis for connexin 43 reached significance only in the mitral valve disease group (p = 0.031). No differences in connexin 40 and connexin 43 expression were detectable between lone AF and AF with mitral valve disease. Within the groups connexin 40 and connexin 43 expression did not differ between patients with paroxysmal AF and patients with chronic AF. CONCLUSION: The present study shows for the first time that AF can induce changes in the left atrium with increased connexin expression. Furthermore, no systematic differences between patients with paroxysmal and chronic AF were detected.

[Exchange mechanism of ion transport in corn roots]. / Austausch-Mechanismus des Ionentransports in Pflanzen am Beispiel des Phosphat- und Chloridtransports bei Maiswurzeln.

The effect of the external anion concentration on the rate of efflux of anions from corn roots was investigated by following the efflux of labeled anions in shaken solutions of different concentrations of unlabeled anions.Rates of efflux of phosphate and chloride from corn roots are rather independent of external concentrations of the anions as long as these concentrations are low.Rates of efflux of phosphate and chloride from corn roots at high external concentrations of these ions were found to be a function of the external concentrations.Stimulation of efflux is specific: phosphate efflux is stimulated only by phosphate and not by chloride in the medium and vice versa. Since it appears unlikely that this effect is due to specific changes in the permeability of the membranes, it may by concluded that the specific stimulation of efflux by external ions as observed in this investigation is caused by ion exchange at mobile membrane units (WEIGL, 1967a). Reasons for this hypothesis are discussed and alternative hypotheses such as that calling for the protoplasm to function as ion exchanger with various specific binding sites are examined. The latter hypothesis remembers to the theory that "the ion carrier is the protoplasm itself" (OVERSTREET, 1957; BANGE and MEIJER, 1966), which diminishes the importance of membranes as limiting boundaries.

[Evidence for mobile transport structures (carriers) involved in ion transport in plants and kinetics of anion transport in elodea in light and dark]. / Beweis für die Beteiligung von beweglichen Transportstrukturen (Trägern) beim Ionentransport durch pflanzliche Membranen und die Kinetik des Anionentransports bei Elodea im Licht und Dunkeln.

Influx and efflux of anions in leaves of Elodea canadensis were investigated. A major component of phosphate efflux was found to be a function of the external phosphate concentration in accord with saturation kinetics, i.e. it was found to be proportional to phosphate influx. This suggests that the transport structures which mediate ion transport in plant membranes are mobile and can work in both directions (Fig. 7C). The higher the external concentration the more bound ions (A b (-) , Fig. 7C) are exchanged for external ions (A a (-) ) instead of being transported back to the inside of the membrane. Thus it was demonstrated that ion transport in plants is mediated by carriers or transportases. The action of the carriers is supported by ATP or another "energy-rich" compound which is regenerated by ATP; ATP may be supplied by the mitochondria or/and chloroplasts. In line with these findings some transport phenomena exhibited features of coenzyme kinetics.Magnitude and molecular nature of the carrier motion are unknown. There is no evidence for rotating or shuttling monovalent carriers. The carriers may be polyvalent and may be part of bigger structural units within the membrane. Nevertheless, these experimental observations are rather consistent with the idea that plant membranes contain tertiary or quarternary formed proteins or lipoproteins than with the conventional lipid bilayer models of cell membranes.

[Efflux and transport of Cl(-) and Rb (+) in corn roots. Action of external KCl, Ca(++), EDTA and IAA]. / Efflux und Transport von Cl(-) und Rb (+) in Maiswurzeln. Wirkung von Außenkonzentration, Ca(++), EDTA und IES.

The efflux of (36)Cl and (86)Rb and the fluxes of these ions into the xylem were investigated using the device shown in Fig. 1.Efflux of (36)Cl is stimulated by external KCl while transport into the xylem is inhibited. Stimulation of the efflux appears to be stronger than inhibition of the transport.The stimulation of the efflux of (36)Cl was also observed with roots of intact seedlings.Assuming that the mode of transfer of Cl(-) into the xylem (flux 3, Fig. 8) is diffusion exhibiting a linear isotherm (LUTTGE and LATIES, 1966), these results suggest that the primary action of external salts is on the efflux across the plasma-lemma (WEIGL, 1967, 1968). We were unable, however, to find a linear relationship between concentration and rate of chloride transport to the shoots of intact seedlings.With respect to the mode of ion transfer to the xylem (WEIGL and LüTTGE, 1965; LUTTGE and LATIES, 1966) we have to be aware of the following facts:A linear isotherm cannot be taken to signify diffusive permeation (TORII and LATIES, 1966; LUTTGE and LATIES, 1966). If the Michaelis constant is extremely high relative to the ion concentration, the relationship between the ion concentration and the rate of a metabolic or mediated transport approaches linearity.The isotherm of the transport into the xylem may primarily reflect the difference of two large fluxes (4 and 5; Fig. 8).The transport data of LUTTGE and LATIES (1966) need not be presented as a straight line (Fig. 6).If at high external ion concentrations the ratio of the ion concentration in the exudation sap to the external ion concentration approaches unity, diffusive permeation into the stele is still not proved to be the mode of migration, since at high stelar ion concentration flux 6 tends to become equal to flux 3.Considerations on radial ion transfer into the xylem depend on contemporary knowledge of the location of transport systems. Cl(-)-uptake into root tips (2 mm) from solutions of 1-10 mM KCl did not exhibit a linear isotherm. These results are unpublished since the discrepancy to the results of TORII and LATIES (1966) may be due to a higher content of vacuoles in our root tips. We feel it unlikely, however, that a linear isotherm of Cl(-)-uptake into root tips is adequately explained by assuming that it is due to a lack of vacuoles while the sensibility to inhibitors is assumed to be due to the presence of vacuoles in root tips.Transport of Cl(-) into the xylem is susceptible to inhibitors of oxydative phosphorylation, suggesting that this process, even at high external ion concentrations, is dependent on metabolic energy in contrast to the passive efflux from the cortical cells across the plasmalemma into the environment of the root. The precise location of the metabolic step(s) on the pathway of ions from the environment of the root to the xylem is unknown.The observed effects of Ca(++), EDTA and IAA may be considered in relation to the theory that auxin exerts its influence on growth by altering the diffusion potential across cell membranes (BRAUNER and DIEMER, 1967). Growth is susceptible to the effect of Ca(++) and EDTA (ADAMSON, 1962; SETTERFIELD, 1963; THIMANN, 1963). Nevertheless, since IAA exerts no influence on ion fluxes in corn roots, it is not clear whether IAA really exerts its influence on growth by altering the diffusion potential across plant cell membranes. We might be dealing with occasional effects of secondary importance.

[Effect of CCCP and UO 2 (++) on ion fluxes in roots]. / Wirkung von CCCP und UO 2 (++) auf die Ionenfluxe in Wurzeln.

Using the device shown in Fig. 1 of a previous paper (Weigl, 1969) efflux of Rb(+) from preloaded corn roots across the plasmalemma of root cortex cells was stimulated by addition of 5×10(-4) M UO 2 (++) , whereas the flux of Rb(+) into the xylem was inhibited by addition of 5×10(-4) M UO 2 (++) . Efflux of Cl(-) across the plasmalemma was stimulated by 10(-5) M CCCP, the flux of Cl(-) via the xylem was inhibited by 10(-5) M CCCP. The problem of interpreting efflux experiments with roots is stated and discussed.

[Fluxes, accumulation and transport of Cl(-) in excised corn roots]. / CL(-)-Fluxe, Akkumulation und Transport in abgeschnittenen Mais-Wurzeln.

Fluxes, accumulation and transport rates of Cl(-) in excised corn roots were investigated.Flux equilibrium in 5×10(-4)M KCl was not reached within an experimental period of 28 hr.Transport of Cl(-) (5×10(-4)M KCl) through the xylem of 4-6 cm long excised corn roots had a lag of 1-2 hr. From 6 to 28 hr rates of accumulation and transport were nearly constant (5×10(-4)M KCl). The velocity of volume-flow within the xylem was 1.5-2 cm hr(-1) (5×10(-4)M KCl). (36)Cl(-)-efflux through the cut end of roots preloaded in K(36)Cl of low concentration exhibits a discontinuity which is explained by addition of two fluxes: efflux of ions concentrated in the xylem (and symplasm) plus efflux from the vacuoles through the xylem.Accumulation of Cl(-) by excised roots approaches a maximum level (Jackson and Edwards, 1966). Influx rates remain constant while efflux rates increase with time. It is shown in this paper that the flux of Cl(-) through the xylem becomes a large proportion of the influx across the plasmalemma. Flux rates suggest that more than 50% of the Cl(-) ions transported to the xylem passed through a vacuole (5×10(-4)M KCl; 20-28 hr).

[Anion influx, ATP level and CO2 fixation in Limnophila gratioloides and Chara foetida]. / Anionen-Influx, ATP-Spiegel und CO2-Fixierung in Limnophila gratioloides und Chara foetida.

Influx of anions (5x10(-4) M Cl(-) or SO4 (2-)) across the plasmalemma, ATP levels and CO2 fixation in Limnophila and Chara have been measured in a comparative study.In Limnophila, influx, ATP level and CO2 fixation were progressively reduced by increasing concentrations of carbonyl cyanide m-chlorophenylhydrazone (CCCP) in the light (4000 lux) as well as in the dark. In Chara, not only influx but also ATP levels were much less reduced in the light than in the dark.At 5x10(-4) M external salt concentration the action of light or dark is to change active influx of anions. Thus this study provides strong evidence to support the view that active anion uptake is directly dependent on ATP rather than on electron flow or NADPH. The possible significance of differences in the photophosphorylation systems of various plants is stressed.

[Purification of the proteinase from Nepenthes pitcher secretion]. / Reinigung der Proteinase aus Nepenthes-Kannensaft.

The proteolytic enzymes from the sterile secretion of the pitchers of 3 Nepenthes species were purified to electrophoretic homogenity by chromatography on Ecteola cellulose. The properties of the purified enzymes were investigated.

Postoperative Mycobacterium avium osteomyelitis confirmed by polymerase chain reaction.

UNLABELLED: An 18-year-old male with Escobar syndrome developed Mycobacterium avium osteomyelitis after corrective osteotomy. After three surgical interventions the infection reappeared a fourth time. Repeated attempts at microbiological diagnosis of the granulomatous lesions by microscopy and culture for conventional bacteria and Mycobacteria did not reveal any organism. The diagnosis of Mycobacterium avium finally was achieved by polymerase chain reaction. Extensive immunological work-up did not reveal signs of immunodeficiency. The patient was treated successfully by a combined surgical and chemotherapeutic approach consisting of clarithromycin, ethambutol and ciprofloxacin. CONCLUSION: Polymerase chain reaction may be especially useful for clinical situations with a low bacterial load, especially for fastidious and slow growing pathogens like Mycobacteria. In our patient a combination of surgical therapy with a triple regimen containing clarithromycin proved successful for treatment of a localised infection with M. avium in a supposedly immunocompetent host.