New insights into the pharmacokinetics and pharmacodynamics of natalizumab treatment for patients with multiple sclerosis, obtained from clinical and in vitro studies.

BACKGROUND: The monoclonal antibody natalizumab (NAT) inhibits the migration of lymphocytes throughout the blood-brain barrier by blocking very late antigen (VLA)-4 interactions, thereby reducing inflammatory central nervous system (CNS) activity in patients with multiple sclerosis (MS). We evaluated the effects of different NAT treatment regimens. METHODS: We developed and optimised a NAT assay to measure free NAT, cell-bound NAT and VLA-4 expression levels in blood and cerebrospinal fluid (CSF) of patients using standard and prolonged treatment intervals and after the cessation of therapy. RESULTS: In paired CSF and blood samples of NAT-treated MS patients, NAT concentrations in CSF were approximately 100-fold lower than those in serum. Cell-bound NAT and mean VLA-4 expression levels in CSF were comparable with those in blood. After the cessation of therapy, the kinetics of free NAT, cell-bound NAT and VLA-4 expression levels differed. Prolonged intervals greater than 4 weeks between infusions caused a gradual reduction of free and cell-bound NAT concentrations. Sera from patients with and without NAT-neutralising antibodies could be identified in a blinded assessment. The NAT-neutralising antibodies removed NAT from the cell surface in vivo and in vitro. Intercellular NAT exchange was detected in vitro. CONCLUSIONS: Incorporating assays to measure free and cell-bound NAT into clinical practice can help to determine the optimal individual NAT dosing regimen for patients with MS.

Coxiella burnetii (Q fever) as a cause of community-acquired pneumonia during the warm season in Germany.

Q fever is a notifiable disease in Germany. The majority of the reported cases are related to outbreaks. The objective of our study was to evaluate the general role of Q fever in community-acquired pneumonia (CAP). We investigated respiratory samples and sera from 255 patients with CAP, who were enrolled into a CAPNETZ cohort in summer 2005. Altogether, our data showed a significant prevalence of Q fever as CAP (3·5%). If a patient's condition leads to a diagnostic test for Chlamydophila sp., Mycoplasma sp. or Legionella sp., then a Q fever diagnostic test should also be included. In particular, ELISA as a first diagnostic step is easy to perform. PCR should be performed at an early stage of the disease if no antibodies are detectable. Because of our highly promising findings we suggest performing PCR in respiratory samples.

Direct observation of nonaffine tube deformation in strained polymer networks.

We present a one-to-one comparison of polymer segmental fluctuations as measured by small angle neutron scattering in a network under deformation with those obtained by neutron spin echo spectroscopy. This allows an independent proof of the strain dependence of the chain entanglement length. The experimentally observed nonaffine square-root dependence of the tube channel on strain is in excellent agreement with theoretical predictions and permits us to exclude an often invoked nondeformed as well as affinely deformed tube.

The urinary antibiotic 5-nitro-8-hydroxyquinoline (Nitroxoline) reduces the formation and induces the dispersal of Pseudomonas aeruginosa biofilms by chelation of iron and zinc.

Since cations have been reported as essential regulators of biofilm, we investigated the potential of the broad-spectrum antimicrobial and cation-chelator nitroxoline as an antibiofilm agent. Biofilm mass synthesis was reduced by up to 80% at sub-MIC nitroxoline concentrations in Pseudomonas aeruginosa, and structures formed were reticulate rather than compact. In preformed biofilms, viable cell counts were reduced by 4 logs at therapeutic concentrations. Complexation of iron and zinc was demonstrated to underlie nitroxoline's potent antibiofilm activity.

Evaluating frequency, diagnostic quality, and cost of Lyme borreliosis testing in Germany: a retrospective model analysis.

BACKGROUND: Data on the economic impact of Lyme borreliosis (LB) on European health care systems is scarce. This project focused on the epidemiology and costs for laboratory testing in LB patients in Germany. MATERIALS AND METHODS: We performed a sentinel analysis of epidemiological and medicoeconomic data for 2007 and 2008. Data was provided by a German statutory health insurance (DAK) company covering approx. 6.04 million members. In addition, the quality of diagnostic testing for LB in Germany was studied. RESULTS: In 2007 and 2008, the incident diagnosis LB was coded on average for 15,742 out of 6.04 million insured members (0.26%). 20,986 EIAs and 12,558 immunoblots were ordered annually for these patients. For all insured members in the outpatient sector, a total of 174,820 EIAs and 52,280 immunoblots were reimbursed annually to health care providers (cost: 2,600,850€). For Germany, the overall expected cost is estimated at 51,215,105€. However, proficiency testing data questioned test quality and standardization of diagnostic assays used. CONCLUSION: Findings from this study suggest ongoing issues related to care for LB and may help to improve future LB disease management.

Established and emerging pathogens in Ixodes ricinus ticks collected from birds on a conservation island in the Baltic Sea.

Tick-borne pathogens such as Lyme borreliosis spirochaetes, Anaplasma phagocytophilum, Rickettsia spp. and Babesia spp. cause a great variety of diseases in animals and humans. Although their importance with respect to emerging human diseases is increasing, many issues about their ecology are still unclear. In spring 2007, 191 Ixodes ricinus (Acari: Ixodidae) ticks were collected from 99 birds of 11 species on a bird conservation island in the Baltic Sea in order to test them for Borrelia spp., A. phagocytophilum, Rickettsia spp. and Babesia spp. infections. Identification of the pathogens was performed by polymerase chain reaction (PCR), restriction fragment length polymorphism and sequence analysis. The majority of birds with ticks testing positive were European robins and thrushes. Borrelia DNA was detected in 14.1%, A. phagocytophilum in 2.6%, rickettsiae in 7.3% and Babesia spp. in 4.7% of the ticks. Co-infections with different pathogens occurred in six ticks (3.1%). The fact that 11 ticks (five larvae, six nymphs) were infected with Borrelia afzelii suggests that birds may, contrary to current opinion, serve as reservoir hosts for this species. Among rickettsial infections, we identified Rickettsia monacensis and Rickettsia helvetica. As we detected five Rickettsia spp. positive larvae and two birds carried more than one infected tick, transmission of those pathogens from birds to ticks appears possible. Further characterization of Babesia infections revealed Babesia divergens and Babesia microti. The occurrence of Babesia spp. in a total of five larvae suggests that birds may be able to infect ticks, at least with Ba. microti, a species considered not to be transmitted transovarially in ticks.

Structure of the Arabidopsis RPM1 gene enabling dual specificity disease resistance.

Plants can recognize pathogens through the action of disease resistance (R) genes, which confer resistance to pathogens expressing unique corresponding avirulence (avr) genes. The molecular basis of this gene-for-gene specificity is unknown. The Arabidopsis thaliana RPM1 gene enables dual specificity to pathogens expressing either of two unrelated Pseudomonas syringae avr genes. Despite this function, RPM1 encodes a protein sharing molecular features with recently described single-specificity R genes. Surprisingly, RPM1 is lacking from naturally occurring, disease-susceptible Arabidopsis accessions.

Adsorption and pinning of multiblock copolymers on chemically heterogeneous patterned surfaces.

The results of Monte Carlo simulations carried out on a system of multiblock copolymers having two different types of monomer units adsorbed on checker board surface configurations are presented here. We investigated the adsorption behavior for a series of different chess board square dimensions and also for various block lengths of the copolymer. We find that the specific heat capacity curves for adsorption indicate double peaks showing a two stage pattern recognition of the copolymer on the surface. It is also seen that the transition that happens at lower temperature corresponds to pinning where the junction point of the different blocks gets pinned to the interface between the different surface sites. It is interesting to see that the multiblock copolymers form multiple pinning sites on the boundary between different kinds of surface sites. There exists an intermediate size of the square on the board where the recognition and pinning are most favored and for smaller and larger size of the board and block length, the adsorption proceeds like in homopolymer on homogeneous surfaces. Unlike in the case of the simplest model of diblock copolymer on stripe-patterned surfaces [K. Sumithra and E. Straube, J. Chem. Phys. 125, 154701 (2006)], here the recognition is stronger and the average adsorption energy and the perpendicular component of the radius of gyration show distinct changes corresponding to the two transitions. The conformational properties of the multiblock copolymer near the checkered surface show interesting effects with the perpendicular component showing strong deviations from the standard behavior.

The Swedish new variant of Chlamydia trachomatis (nvCT) remains undetected by many European laboratories as revealed in the recent PCR/NAT ring trial organised by INSTAND e.V., Germany.

The May 2009 round of INSTAND's ring trial "Chlamydia trachomatis detection PCR/NAT" included a sample with high amount of the Swedish new variant of C. trachomatis (nvCT). A spectrum of at least 12 different commercial diagnostic nucleic acid amplification tests (NAATs) and many different in house NAATs were applied by the 128 participating laboratories which reported 152 results. Approximately 80% of the results correctly reported the presence of C. trachomatis in the nvCT specimen. The nvCT sample was mainly missed, as expected, by participants using the Roche COBAS Amplicor CT/NG (15.5% of reported results) but also by several participants using in house NAATs. The trend towards using nvCT-detecting NAATs is obvious and in addition to the new dual-target NAATs from Roche and Abbott, and BD ProbeTec ET, also a number of new CE mark-certified commercial tests from smaller diagnostic companies as well as many different in house NAATs were used. Laboratories using commercial or in house NAATs that do not detect the nvCT are encouraged to carefully monitor their C. trachomatis incidence, participate in appropriate external quality assurance and controls schemes, and consider altering their testing system. The reliable detection of low amounts of the wildtype C. trachomatis strain in other samples of the ring trial set indicates a good diagnostic performance of all applied commercial NAATs while also detecting the nvCT strain.

Low-grade infections as a possible cause of arthrofibrosis after total knee arthroplasty.

PURPOSE: Arthrofibrosis after total knee arthroplasty represents a considerable burden for the patient and a therapeutic challenge for the practitioner. One possible cause discussed in the literature is a low-grade infection. This hypothesis should be examined within the scope of this retrospective study. PATIENTS AND METHODS: Nineteen patients with clinical symptoms of arthrofibrosis after primary total knee arthroplasty were examined between January, 1999 and January, 2012. Incorrect positioning was radiologically ruled out. All patients were examined clinically (score of Freeman as well as Blauth and Jäger), radiologically (component and leg alignment, patella height according to Insall and Salvati), microbiologically (culture-based procedures), molecular biologically (PCR) and histologically in the course of an open revision of the prosthesis. RESULTS: According to the score of Freeman et al. (1977), a highly significant improvement in pain (p = 0.007) and in the overall score (p = 0.003) was shown. The knee joint mobility did not change significantly (p = 0.795). PCR was negative in 17 patients. One patient showed a PCR-positive result of the synovial membrane for Corynebacterium spp., while Staphylococcus warneri was detected in the culture. Another patient had a positive result of synovia PCR for Enterococcus cecorum as well as Corynebacterium spp. However, this culture was sterile. In 16 patient samples, no bacterial growth was detectable. Two samples were not evaluable. The main histopathological findings were synovialitis and fibrosis. CONCLUSION: The hypothesis of low-grade-infection-induced arthrofibrosis after total knee arthroplasty could not be confirmed in this study. However, based on this small study population the conclusion needs to be confirmed by new and larger studies, ideally prospectively designed including a control group.

Costs and quality of life of multiple sclerosis in Germany.

This cost-of-illness analysis based on information from 2973 patients with multiple sclerosis (MS) in Germany is part of a Europe-wide study on the costs of MS. The objective was to analyze the costs and quality of life (QOL) related to the level of disease severity. Patients from six centres (office- and hospital-based physicians) and patients enrolled in a database were asked to participate in the survey; 38% answered a mail questionnaire. In addition to details on the disease (type of disease, relapses, level of functional disability), the questionnaire asked for information on all resource consumption, medical, non-medical, work absence, informal care, as well as QOL (measured as utility). The mean age of the cohort was 45 years, and 18% of patients were 65 years of age or older. Forty-seven percent of patients had mild disease (Expanded Disability Status Scale [EDSS] score 0-3), 36% had moderate disease (EDSS score 4-6.5) and 12% had severe disease (EDSS score > or =7). The mean EDSS score in the sample was 3.8 (median 4.0), with a mean utility of 0.62. Costs and utility are highly correlated with disease severity. Workforce participation decreases from 73% in very early disease to less than 10% in the very late stages, leading to a tenfold rise in productivity losses in the late stages of disease. Hospitalisation and ambulatory visits rise by a factor of 5-6 between early and late disease; investments and services increase from basically no cost to euro 2700; and informal care increases by a factor of 27 for patients with an EDSS score of 7 and by a factor of 50 for patients at the very severe end of the EDSS scale (8-9). Hence, total mean costs per patient are determined essentially by the distribution of the severity levels in the sample, increasing from approximately euro 18 500 at an EDSS score of 0-1 to euro 70 500 at an EDSS score of 8-9. The same is true for utility, which decreases from 0.86 to 0.10 as the disease becomes severe. However, the utility loss compared to the general population is high at all levels of the disease, leading to an estimated loss of 0.2 quality-adjusted life-years per patient. Relapses are associated with a cost of approximately euro 3 000 and a utility loss of 0.1 during the quarter in which they occur. Compared with a similar study performed in 1999, resource consumption, with the exception of drugs, is somewhat lower. This is most likely due to a difference in the severity distribution of the two samples and to changes in health-care consumption overall in the country, such as the introduction of diagnosis-related groups (DRGs, Fallpauschalen).

Insulin-like growth factor 1 and a cytosolic tyrosine kinase activate chloride outward transport during maturation of hippocampal neurons.

The development of hyperpolarizing inhibition is an important step in the maturation of neuronal networks. Hyperpolarizing inhibition requires Cl(-) outward transport that is accomplished by KCC2, a K(+)/Cl(-) cotransporter. We show that cultured hippocampal neurons initially contain an inactive form of the KCC2 protein, which becomes activated during subsequent maturation of the neurons. We also show that this process is accelerated by transient stimulation of IGF-1 receptors. Because the transporter can be rapidly activated by coapplication of IGF-1 and an Src kinase and can be deactivated by membrane-permeable protein tyrosine kinase inhibitors, we suggest that activation of K(+)/Cl(-) cotransporter function by endogenous protein tyrosine kinases mediates the developmental switch of GABAergic responses to hyperpolarizing inhibition.

In situ hybridisation and direct fluorescence antibodies for the detection of Chlamydia trachomatis in synovial tissue from patients with reactive arthritis.

BACKGROUND: Chlamydia trachomatis is associated with Reiter's syndrome and reactive arthritis but the form in which the organism survives in synovial cells is unclear. AIM: To compare in situ hybridisation with direct fluorescence in the detection of inapparent chlamydial infection in synovial tissue. METHODS: Synovial tissue from four patients with reactive arthritis patients was examined using biotin labelled probes for chlamydial DNA and fluorescein isothiocyanate (FITC) labelled monoclonal antibodies against the major outer membrane protein. RESULTS: In two of the four patients, evidence of chlamydial infections was detected by in situ hybridisation in parallel sections but not with FITC labelled monoclonal antibodies. CONCLUSIONS: Detection of chlamydial DNA by in situ DNA hybridisation may be a better way to identify chlamydial infection in synovial tissue than phenotype targeting with FITC conjugated antibodies, which is used as a standard procedure for screening clinical specimens for chlamydia.

The effects of inhibiting leukocyte migration with fucoidin in a rat peritonitis model.

OBJECTIVES: To study the effects of fucoidin on leukocyte rolling and emigration and bacterial colonization in a peritonitis sepsis model in rats. DESIGN AND INTERVENTIONS: A controlled study in 64 male Wistar rats, anesthetized and rendered septic by cecal ligation and puncture (CLP). Immediately after CLP 32 animals received a continuous infusion of fucoidin and 32 a continuous infusion of Ringer's lactate. MEASUREMENTS AND MAIN RESULTS: Systemic leukocyte counts were determined every 2 h after CLP. Surviving animals were anesthetized 24 h after CLP, and intravital measurements of leukocyte rolling in venules in the cremaster muscle were performed. The animals were then killed and their organs harvested for histological and microbiological examinations. The 24-h survival was comparable in the two groups. Fucoidin-treated animals had higher leukocyte counts in the systemic circulation and lower counts in the lungs, liver, abdominal cavity, and brain than control animals. The number of bacterial colony forming units in the abdominal cavity, lungs, liver, brain and blood did not differ in the two groups. Fucoidin treatment changed the type of bacteria predominantly found in the examined organs from Escherichia coli to Pseudomonas aeruginosa. CONCLUSIONS: In an intra-abdominal model of sepsis we found that treatment with fucoidin induces leukocytosis inhibits leukocyte rolling and reduces leukocyte emigration in the abdominal cavity, lungs, and liver. Reduction in the number of emigrating leukocytes was not associated with an increase in bacterial counts found in the examined organs.

Interferon-beta induction by Chlamydia pneumoniae in human smooth muscle cells.

Clinical studies have suggested a causal or contributory role of Chlamydia pneumoniae infection in asthma and atherosclerosis. The activation of synthetic functions of smooth muscle cells (SMC) including the production of cytokines and growth factors plays a major role in the formation of fibrous atherosclerotic plaques as well as in structural remodelling of the airway wall in chronic asthma. In this study we demonstrated that C. pneumoniae induced the production of low levels of interferon (IFN)-beta in bronchial and vascular SMC when infected cells were treated with tumour necrosis factor-alpha (TNF-alpha). IFN-beta production was analysed by reverse transcription-PCR and enzyme-linked immunosorbent assay. The upregulation of IFN-beta was paralleled by an increase in mRNA levels of interferon regulatory factor-1 and interferon-stimulated gene factor 3gamma, two transcription factors activating the expression of the IFN-beta gene. In addition, C. pneumoniae infection enhanced the mRNA level of indoleamine 2,3-dioxygenase, an IFN-inducible factor mediating the restriction of intracellular chlamydial growth, in TNF-alpha-stimulated SMC. C. pneumoniae-induced IFN-beta production by SMC may modulate inflammation and tissue remodelling during respiratory and vascular infection.

M protein of a Streptococcus dysgalactiae human wound isolate shows multiple binding to different plasma proteins and shares epitopes with keratin and human cartilage.

Besides group A (GAS), Lancefield group C beta-haemolytic streptococci (GCS) have been implicated as a causative agent in outbreaks of purulent pharyngitis. In this study we have investigated a class CI M protein of a Streptococcus dysgalactiae1:256, revealed that 26% of these sera showed serological cross-reactivity between a 68-kDa cartilage protein and the N-terminal part of MC. Only 8% of the sera of healthy patients showed this property. In additional, MC also cross-reacted with antibodies recognising epidermal keratins. The cross-reacting 68-kDa protein from cartilage was different from human serum albumin, but was recognised with anti-vimentin immune serum. The MC was cloned and the gene sequenced. By using PCR, recombinant gene fragments encoding characteristic peptide fragments of MC were expressed in Escherichia coli. The peptides were used to map the binding sites for plasma proteins and to locate the cross-reacting epitopes on the MC molecule. In consequence, sequence alignments revealed that MC shared homologous regions with vimentin and different keratins. Our data, obtained with MC, suggest that not only infections with GAS but also infections with GCS and possibly GGS (the latter species can also produce class CI M-like proteins) may be responsible for the formation of streptococcal-associated sequel diseases.

Findings significant with respect to short- and medium-term outcome in schizophrenia--a preliminary report.

1. The objective of the study was to determine the relationship of psychopathological, psychobiological, and clinical psychological variables to short- and medium-term outcome in schizophrenia. 2. The predictive power of several variables described as potential predictors in the literature were evaluated. To these we added clinical psychological variables; most of these had never been examined in a prospective study. 3. The sample consisted of a total of 45 patients. None of the patients was on medication and all were tested shortly after admission to the hospital. The following factors were regarded as outcome criteria: change in the GAS-score four weeks after admission, length of hospitalization after index admission, relapse nine months after admission and two years after discharge. 4. We found that the psychobiological data set as well as the clinical psychological data set had the highest predictive power with respect to short- and medium-term outcome. 5. An analysis of the single variables within the clinical psychological data set revealed that the patients with attention disorder and a low Stimulus Barrier Function (Bellak-scale) responded better to a four-week Haloperidol treatment. Patients with a low Stimulus Barrier Function at admission had, in comparison to this, a poor outcome after two years (relapse). (No patient was taking drugs regularly at this time.) The expressed emotion of the relative approached only significance (p less than .07) as a factor to be connected with relapse after two years. 6. As psychological factors the ANS activity and the CT were measured. The latter in order to evaluate possible atrophic alterations in the brain. 7. Higher activity of the ANS was found in those patients who were hospitalized for a longer period. Slight atrophic alterations of the brain (CT scan) were related to poor outcome after two years (relapse). 8. No significant relationship was found between the psychopathological state at admission and short- or medium-term course of the illness. 9. The findings are interpreted as showing the importance of psychobiological as well as clinical psychological variables in order to uncover the factors related to the heterogeneous course of the schizophrenic illness.

Antimicrobial susceptibility of anaerobic and capnophilic bacteria isolated from odontogenic abscesses and rapidly progressive periodontitis.

In dentistry antimicrobials are used in the treatment of progressive periodontitis and odontogenic abscesses, therefore the susceptibility to commonly used antibiotics of capnophilic and anaerobic species causing these diseases should be investigated. The activity of penicillin, amoxycillin, cefoxitin, clindamycin, doxycycline, metronidazole and ciprofloxacin was investigated. One hundred and sixty four isolates from subgingival plaque samples of 66 patients with progressive periodontitis and 192 bacterial strains from pus of 74 patients with odontogenic abscesses were included in this study. The majority of species tested were gram-negative anaerobes (Prevotella spp., Porphyromonas spp., Fusobacterium spp.), and were highly susceptible to clindamycin and metronidazole. Nearly 6% of the periodontal isolates and 22% of the bacteria obtained from pus samples produced beta-lactamases. With the exception of the periodontopathogenic species Actinobacillus actinomycetemcomitans and Eikenella corrodens, clindamycin seemed to be a useful antibiotic and could be recommended for empirical antimicrobial treatment.

Secretion of cytokines by uroepithelial cells stimulated by Escherichia coli and Citrobacter spp.

Urinary tract epithelial cells (T 24/83) are able to express interleukin (IL)-6, IL-8, platelet-derived growth factor (PDGF) and tumour necrosis factor-alpha, but not IL-1 beta, IL-2, IL-4 and IL-10 in response to an infection with uropathogenic bacteria. The process of cytokine secretion is time dependent, with a significant increase in the cytokine activity after 60 min. The expression of virulence factors of the bacteria does not seem to play a role. The interaction between bacterial products (e.g. lipopolysaccharide) and/or bacterial adhesion mediated by adhesins and specific receptor molecules of cell surfaces may be responsible for the activity of mediator protein expression in the epithelial cells. The release of PDGF and IL-8 was found to be higher when due to Escherichia coli HB 101 (rough form) than that caused by other bacterial strains. Citrobacter CB 3009 provoked the highest level of IL-6. The PDGF level correlated significantly with IL-6 and IL-8 values (P<0.001). There was a significant correlation between the time-dependent release of IL-6 and IL-8 (P<0.05). In epithelial cytokine response to bacterial infection, the reaction of the epithelial cells may modify themselves (e.g. internalization of bacteria) and the immuno-regulatory processes that are caused by infection and responsible for parenchymal injury.

The influence of ofloxacin (Tarivid) on the parasite-host inter-relationship in patients with chronic urinary tract infection.

In urinary tract infection (UTI), the aim of antimicrobial chemotherapy is either to attenuate the virulence of infective micro-organisms, to influence the interaction between germs and host or to kill bacteria. In 25 females (aged 49.8+/-14.7 years) with chronic UTI, parameters of inflammation (CRP, alpha 2-globulin, leukocytes, ESR) as well as renal function were analyzed under the treatment with ofloxacin 2 x 200 mg. Subsequently, bacterial attachment, bacterial count, leukocytes, antibody-coated bacteria (ACB), immunoglobulin (Ig) G, albumin, a2-microglobulin, Tamm Horsfall protein, secretory IgA (sIgA) and lysozyme were determined in urine. The micro-organisms were examined with regard to the expression of hemolysin, aerobactin, P-fimbriae and according to their plasmid profile. Ofloxacin serum levels were analyzed once prior to, on day 6 during and on day 3 after drug administration. In all cases, the acute clinical symptoms had disappeared after 10 days of treatment, all bacteria were eliminated, and the parameters of inflammation in serum and urine had returned to normal. On the sixth day of therapy, no expression of P-fimbriae was detectable in the Escherichia coli strains isolated, and the attachment rate decreased from 42+/-30.9% to 11.1+/-18.1%. The sIgA level rose from 42.6+68.5 prior to therapy to 88.8+/-136.8 mmol/l on day 3 after therapy; acute symptoms of UTI did not recur in any case during the period of 1 year.