Effectiveness of Different Antimicrobial Strategies for Staphylococcal Prosthetic Joint Infection: Results From a Large Prospective Registry-Based Cohort Study.

Background: Treatment of staphylococcal prosthetic joint infection (PJI) usually consists of surgical debridement and prolonged rifampicin combination therapy. Tailored antimicrobial treatment alternatives are needed due to frequent side effects and drug-drug interactions with rifampicin combination therapy. We aimed to assess the effectiveness of several alternative antibiotic strategies in patients with staphylococcal PJI. Methods: In this prospective, multicenter registry-based study, all consecutive patients with a staphylococcal PJI, treated with debridement, antibiotics and implant retention (DAIR) or 1-stage revision surgery between January 1, 2015 and November 3, 2020, were included. Patients were treated with a long-term rifampicin combination strategy (in 2 centers) or a short-term rifampicin combination strategy (in 3 centers). Antimicrobial treatment strategies in these centers were defined before the start of the registry. Patients were stratified in different groups, depending on the used antimicrobial strategy. Cox proportional hazards models were used to compare outcome between the groups. Results: Two hundred patients were included and stratified in 1 long-term rifampicin group (traditional rifampicin combination therapy) or 1 of 3 short-term rifampicin groups (clindamycin or flucloxacillin or vancomycin monotherapy, including rifampicin for only 5 postoperative days). Adjusted hazard ratios (aHRs) for failure in patients treated with short-term rifampicin and either flucloxacillin or clindamycin were almost equal to patients treated with long-term rifampicin combination therapy (aHR = 1.21; 95% confidence interval, .34-4.40). Conclusions: A short-term rifampicin strategy with either clindamycin or flucloxacillin and only 5 days of rifampicin was found to be as effective as traditional long-term rifampicin combination therapy. A randomized controlled trial is needed to further address efficacy and safety of alternative treatment strategies for staphylococcal PJI.

Floating marine macro litter in the Black Sea: Toward baselines for large scale assessment.

The Black Sea is a semi-enclosed basin subject to major anthropogenic pressures, including marine litter and plastic pollution. Due to numerous large rivers draining into the basin and a population settled along the coast, the region could accumulate significant amounts of floating litter over time. Until now, only limited field data were available, and litter quantities and distribution remained unknown. In this study, floating marine macro litter (FMML) was assessed at the regional Black Sea scale for the first time, showing relatively high litter densities across the basin that reached a weighted mean of 81.5 items/km2. Monitoring data revealed an accumulation of floating items offshore in the eastern part of the basin, resembling on a small scale a 'garbage patch', where litter items were trapped, showing elevated densities in comparison to their surrounding areas. Most of these items were made of plastic materials (ca. 96%) and included large numbers of plastic and polystyrene fragments of small size ranges (2.5-10 cm). Harmonised field data collection through consistent and regular monitoring programmes across the region is essential to establish baselines and thresholds for large scale assessment at international level.

Unpredictable episodic breathlessness in patients with advanced chronic obstructive pulmonary disease and lung cancer: a qualitative study.

PURPOSE: The internationally consented definition and categorization describe two categories of episodic breathlessness: predictable (with known triggers) and unpredictable. The link of known triggers only to predictable episodes can be read that unpredictable episodes have none known trigger. Our aim was to illuminate patients' experiences with episodes of unpredictable breathlessness, to collect descriptions of the episodes' impact on the patients' lives, and, in turn, the patients' individual coping strategies in this connection. DESIGN: Qualitative study using semi-structured in-depth interviews with patients suffering from unpredictable episodes of breathlessness and chronic obstructive pulmonary disease (COPD; Global Initiative for Obstructive Lung Disease III and IV) or lung cancer (all stages). Interviews were audio-recorded, transcribed verbatim, and analyzed using Framework Analysis. RESULTS: One hundred one patients were screened in a large university hospital; ten participants fulfilled the inclusion criteria and provided consent. The experienced episodes were evaluated as unpleasant and with higher intensity compared to predictable episodes. Non-pharmacological interventions were identified as useful coping strategies. Interestingly, although patients experienced the episodes in an unpredictable manner, a trigger could be detected retrospectively for the majority of cases (mostly emotions (especially panic) and, occasionally, physical exertion). Unpredictable episodes are less frequent than previously assumed. CONCLUSION: The unpredictability of unpredictable breathless episodes refers to the patients' experience that these episodes occur "out-of-the-blue." However, a known trigger can be identified for the majority of unpredictable breathless episodes. These are therefore triggered as well. Further research needs to describe more possible triggers, to inquire the prevalence of unpredictable episodic breathlessness, and to develop effective management strategies.

Pre-emptive treatment with fibrinogen concentrate for postpartum haemorrhage: randomized controlled trial.

BACKGROUND: In early postpartum haemorrhage (PPH), a low concentration of fibrinogen is associated with excessive subsequent bleeding and blood transfusion. We hypothesized that pre-emptive treatment with fibrinogen concentrate reduces the need for red blood cell (RBC) transfusion in patients with PPH. METHODS: In this investigator-initiated, multicentre, double-blinded, parallel randomized controlled trial, we assigned subjects with severe PPH to a single dose of fibrinogen concentrate or placebo (saline). A dose of 2 g or equivalent was given to all subjects independent of body weight and the fibrinogen concentration at inclusion. The primary outcome was RBC transfusion up to 6 weeks postpartum. Secondary outcomes were total blood loss, total amount of blood transfused, occurrence of rebleeding, haemoglobin <58 g litre(-1), RBC transfusion within 4 h, 24 h, and 7 days, and as a composite outcome of 'severe PPH', defined as a decrease in haemoglobin of >40 g litre(-1), transfusion of at least 4 units of RBCs, haemostatic intervention (angiographic embolization, surgical arterial ligation, or hysterectomy), or maternal death. RESULTS: Of the 249 randomized subjects, 123 of 124 in the fibrinogen group and 121 of 125 in the placebo group were included in the intention-to-treat analysis. At inclusion the subjects had severe PPH, with a mean blood loss of 1459 (sd 476) ml and a mean fibrinogen concentration of 4.5 (sd 1.2) g litre(-1). The intervention group received a mean dose of 26 mg kg(-1) fibrinogen concentrate, thereby significantly increasing fibrinogen concentration compared with placebo by 0.40 g litre(-1) (95% confidence interval, 0.15-0.65; P=0.002). Postpartum blood transfusion occurred in 25 (20%) of the fibrinogen group and 26 (22%) of the placebo group (relative risk, 0.95; 95% confidence interval, 0.58-1.54; P=0.88). We found no difference in any predefined secondary outcomes, per-protocol analyses, or adjusted analyses. No thromboembolic events were detected. CONCLUSIONS: We found no evidence for the use of 2 g fibrinogen concentrate as pre-emptive treatment for severe PPH in patients with normofibrinogenaemia. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov: http://clinicaltrials.gov/show/NCT01359878. Published protocol: http://www.trialsjournal.com/content/pdf/1745-6215-13-110.pdf.

Identification of N-terminal regions of wheat leaf ferredoxin NADP+ oxidoreductase important for interactions with ferredoxin.

Wheat leaves contain two isoproteins of the photosynthetic ferredoxin:NADP(+) reductase (pFNRI and pFNRII). Truncated forms of both enzymes have been detected in vivo, but only pFNRII displays N-terminal length-dependent changes in activity. To investigate the impact of N-terminal truncation on interaction with ferredoxin (Fd), recombinant pFNRII proteins, differing by deletions of up to 25 amino acids, were generated. During purification of the isoproteins found in vivo, the longer forms of pFNRII bound more strongly to a Fd affinity column than did the shorter forms, pFNRII(ISKK) and pFNRII[N-2](KKQD). Further truncation of the N-termini resulted in a pFNRII protein which failed to bind to a Fd column. Similar k(cat) values (104-140 s(-1)) for cytochrome c reduction were measured for all but the most truncated pFNRII[N-5](DEGV), which had a k(cat) of 38 s(-1). Stopped-flow kinetic studies, examining the impact of truncation on electron flow between mutant pFNRII proteins and Fd, showed there was a variation in k(obs) from 76 to 265 s(-1) dependent on the pFNRII partner. To analyze the sites which contribute to Fd binding at the pFNRII N-terminal, three mutants were generated, in which a single or double lysine residue was changed to glutamine within the in vivo N-terminal truncation region. The mutations affected binding of pFNRII to the Fd column. Based on activity measurements, the double lysine residue change resulted in a pFNRII enzyme with decreased Fd affinity. The results highlight the importance of this flexible N-terminal region of the pFNRII protein in binding the Fd partner.

A screen for potential ferredoxin electron transfer partners uncovers new, redox dependent interactions.

Ferredoxin (Fd) is the primary soluble acceptor at the end of the photosynthetic electron transport chain, and is known to directly transfer electrons to a wide range of proteins for use in metabolism and regulatory processes. We have conducted a screen to identify new putative Fd interaction partners in the cyanobacteria Synechocystis sp. PCC 6803 using Fd-chromatography in combination with MALDI-TOF mass spectrometry. Many novel interactions were detected, including several redox enzymes, which are now candidates for further experiments to investigate electron transfer with Fd. In addition, some proteins with regulatory activity related to photosynthesis were identified. We cloned and expressed one such protein, known as RpaA, which is a specific regulator of energy transfer between phycobilisomes and PSI. Using the recombinant protein we confirmed direct interaction with Fd, and discovered that this was dependent on redox state. The screen for putative Fd-binding proteins was repeated, comparing oxidizing and reducing conditions, identifying many proteins whose interaction with Fd is redox dependent. These include several additional signaling molecules, among them the LexA repressor, Ycf53 and NII, which are all involved in interpreting the redox state of the cell.

Prospective pilot intervention study to prevent medication errors in drugs administered to children by mouth or gastric tube: a programme for nurses, physicians and parents.

BACKGROUND: Drug administration in children is an error-prone task for nurses and parents because individual dose adjustment is often necessary, and suitable formulations for children are frequently lacking. Hence, in the absence of measures for their prevention, medication errors are likely to occur. OBJECTIVE: To assess the error prevalence in drug administration by mouth or gastric tube before and after implementing a programme for quality improvement for nurses and parents. DESIGN, SETTING AND PARTICIPANTS: Prospective, two-period cohort intervention study on a paediatric neurology ward of a university hospital where drug administration procedures of nurses and parents were consecutively monitored during the routine drug administration hours. MAIN OUTCOMES MEASURE: Prevalence of administration errors before and after implementing instructions for appropriate drug administration, and a teaching and training programme supported by information pamphlets. RESULTS: Altogether, 1164 predefined administration tasks were assessed, 675 before and 489 after the intervention. Of these, 95.7% (after the INTERVENTION: 92.6%) were performed by nurses. Errors addressed by the intervention were reduced from 261/646 tasks (40.4%) to 36/453 (7.9%, p<0.001) in nurses and from 28/29 (96.6%) to 2/36 (5.6%, p<0.001) in parents. Errors in predefined categories concerning tablet dissolution, tablet storage, oral liquids, tablet splitting, administration by gastric tube and others were all considerably less frequent after the intervention (each p<0.001). CONCLUSION: Errors of drug administration by mouth and gastric tube represent a considerable and often neglected drug-related problem in paediatric inpatients. Targeted quality-improvement programmes can substantially and rapidly reduce error prevalence. Appropriate teaching and training of both nurses and parents supported by pamphlets was a highly efficient way to reduce error prevalence.

Atmospheric input of POPs into Lake Maggiore (Northern Italy): PCDD/F and dioxin-like PCB profiles and fluxes in the atmosphere and aquatic system.

The delivery and accumulation of PCDD/Fs and dioxin-like PCBs (DL-PCBs) as recorded in sediments from the Lake Maggiore (LM) in Northern Italy and the influence of the atmosphere in contaminating this subalpine ecosystem were studied. PCDD/Fs (17 congeners) and DL-PCBs (12 congeners) concentrations, congener profiles and fluxes in air, bulk deposition, aquatic settling material and surface sediments are presented. Our strategy was to obtain surface sediment from areas near riverine inputs and in the depositional basins, settling material in sediment traps at one site and air, aerosol and bulk deposition profiles. PCDD/F concentrations in air of 25 WHO-TEQ fg m(-3) and DL-PCBs of 7 WHO-TEQ fg m(-3) were measured. Bulk atmospheric deposition yielded 140 and 28 WHO-TEQ pg m(-2) per week for dioxins/furans and DL-PCBs, respectively. Aquatic settling material exhibited concentrations of 11 WHO-TEQ pg g(-1) for PCDD/Fs and 2 WHO-TEQ pg g(-1) for DL-PCBs. Weekly settling material fluxes of 50 WHO-TEQ pg m(-2) and 10 WHO-TEQ pg m(-2) were obtained for PCDD/Fs and DL-PCBs, respectively. PCDD/Fs concentrations in surface sediments varied from 0.1 to 17 WHO-TEQ pg g(-1) whereas values ranged from 0.03 to 6 WHO-TEQ pg g(-1) for DL-PCBs. The concentrations obtained in the environmental compartments studied suggest a situation of low level contamination. PCDD/Fs and DL-PCBs congener profiles in atmospheric air, bulk deposition, aquatic settling matter and surface sediment point to an important if not dominant contribution from atmospheric deposition to Lake Maggiore, especially derived from wet deposition of aerosol-bound PCDD/Fs and DL-PCBs. Flux estimates support this hypothesis.

Atmospheric input of POPs into Lake Maggiore (Northern Italy): PBDE concentrations and profile in air, precipitation, settling material and sediments.

Large lakes are sinks for many chemical pollutants but the role of the atmosphere in delivering PBDEs (polybrominated diphenyl ethers) is not well known. In this study we estimated inputs of PBDEs from the atmosphere to Lake Maggiore and the delivery via settling material to accumulated bottom sediments. Sampling consisted of one simultaneous week of air and bulk deposition during Spring 2005, an integrated 4-month period of collected settling material (via sediment trap), and superficial bottom sediments near the sediment trap deployment and tributary mouths of the lake. Concentration of total PBDEs (Sigma 8) in the sediments ranged between 0.06 and 27 ng g(-1), and two different patterns were observed. One pattern was dominated by BDE-47 and BDE-99 and a second by BDE-209. The latter pattern suggested input from local source(s). Total PBDEs in air were 107 pg m(-3) with a pattern dominated by BDE-47. The bulk deposition rate of total PBDEs for precipitation was measurated to be 17.6 ng m(-2) day(-1) where BDE-209 exhibited the highest concentration. For aquatic settling material the flux was 3.57 ng m(-2) day(-1) and was dominated by congeners BDE-47 and BDE-99. The bulk deposition and settling material fluxes were in the same range for the less brominated congeners, while for the hepta- and deca-brominated PBDEs the fluxes in the settling material were one order of magnitude lower. This suggests different sedimentation processes among congeners and/or the presence of local sources that influence the relative distribution in water column.

[Proliferation and differentiation of human osteoblasts from the nasal septum in a new perfusion culture system]. / Vermehrung und Differenzierung humaner Nasenseptum-Osteoblasten. Ein neuartiges Perfusionskultursystem.

BACKGROUND: Today, perfusion culture systems are mainly used to investigate cellular physiology and to cultivate three-dimensional tissue complexes. As a rule, these systems are relatively expensive and do not enable continuous microscopic monitoring of the growing cells. Simple and inexpensive perfusion culture systems have not been available up to now. METHODS: A novel perfusion culture system was developed in which the modular components consist of a mounting apparatus for inserting various media supply systems, microdispenser pumps, and laminar-flow culture chambers, each with a culture volume of 8 cm(3). The perfusion chambers were inoculated with human osteoblast cells from the tissue culture (5,000/cm(2)) and were perfused for 10 days after adherence of the cells (0.5 ml/min). As a control group, osteoblast-like cells were cultured in identically constructed culture chambers without medium perfusion. After 10 days, the cell counts were determined in accordance with the Coulter principle. Alkaline phosphatase was measured photometrically as a characteristic for differentiation. RESULTS: Compared with the control group, three to four times the quantity of cells were produced within 10 days in the perfusion cultures. The alkaline phosphatase values were equally high or only slightly lower, indicating that osteoblast differentiation of the cells was maintained with a higher proliferation. CONCLUSIONS: As large a number of in vitro proliferated cells as possible is a prerequisite for clinical application of tissue engineering. By continuously supplying medium, the tested perfusion culture system enables a higher rate of proliferation of osteoblast-like cells with maintenance of differentiation. Continuous microscopic monitoring of the cultures is possible using commercially available Petri dishes.

Expression of multiple forms of ferredoxin NADP+ oxidoreductase in wheat leaves.

In higher plants there are two forms of ferredoxin NADP(+) oxidoreductase (FNR), a photosynthetic pFNR primarily required for the photoreduction of NADP(+), and a heterotrophic hFNR which generates reduced ferredoxin by utilizing electrons from NADPH produced during carbohydrate oxidation. The aim of this study was to investigate the presence of multiple forms of FNR in wheat leaves and the capacity of FNR isoforms to respond to changes in reductant demand through varied expression and N-terminal processing. Two forms of pFNR mRNA (pFNRI and pFNRII) were expressed in a similar pattern along the 12 cm developing primary wheat leaf, with the highest levels observed in plants grown continuously in the dark in the presence (pFNRI) or absence (pFNRII) of nitrate respectively. pFNR protein increased from the leaf base to tip. hFNR mRNA and protein was in the basal part of the leaf in plants grown in the presence of nitrate. FNR activity in plants grown in a light/dark cycle without nitrate was mainly due to pFNR, whilst hFNR contributed significantly in nitrate-fed plants. The potential role of distinct forms of FNR in meeting the changing metabolic capacity and reductant demands along the linear gradient of developing cells of the leaf are discussed. Furthermore, evidence for alternative N-terminal cleavage sites of pFNR acting as a means of discriminating between ferredoxins and the implications of this in providing a more effective flow of electrons through a particular pathway in vivo is considered.

Seasonal variations of selected herbicides and related metabolites in water, sediment, seaweed and clams in the Sacca di Goro coastal lagoon (Northern Adriatic).

This study assesses the status of Sacca di Goro coastal lagoon (Northern Adriatic, Italy) with respect to watershed pollution. Because 80% of its watershed is devoted to agriculture, plant protection products and their metabolites were found in the water column, sediments (the upper 0-15 cm layer), macroalgae (Ulva rigida) and clams (Tapes philippinarum). Five seasonal sampling campaigns were performed from May 2004 to April 2005 and concentrations measured in five stations in the lagoon and six in the watershed. Relatively high concentrations of the s-triazine - terbuthylazine -, urea herbicides - diuron - and alachlor were detected through the year mainly at stations directly influenced by the Po di Volano inflow. The concentrations of products in use follow a clear seasonal pattern with spring peaks. This pattern is also visible in the sediments as well as in biota. Among metabolites, hydroxylated compounds prevailed, often with concentrations greater than those of the parent compounds. For the most part of the year, the concentrations in biota were close to detection limits, with concentration peaks in spring.

[Significance of voice constitution as a predisposition for occupational voice disorders]. / Bedeutung der Stimmkonstitution für die Entstehung von Berufsdysphonien.

Occupational voice disorders have been increasing for years. The aim of this study was to examine whether a constitutionally weak voice should be regarded as a risk factor for developing such voice disorders. In a prospective study, 15 female teacher students with a normal vocal constitution were compared with 18 students with constitutionally weak voices during teaching practice. There was a significant difference in the mean fundamental frequency of both groups after teaching for 30 min. Students with constitutionally weak voices tended to have increased values. Taking into consideration the physiological aspects of an increasing fundamental frequency as a sign of vocal fatigue, a constitutionally weak voice has to be regarded as a potential risk factor for developing a voice disorder in vocally intensive occupations. To prevent future vocal problems by appropriate prophylactic intervention (i.e. voice therapy, introduction to vocal hygiene), a phoniatric examination of vocal constitution and endurance is recommended at the beginning of a voice dependent professional career.

[Realization of a measurement module for determining pH value in perfusion cultures]. / Realisierung eines Messmoduls für die pH-Wert-Bestimmung in Perfusionskulturen.

For cultivating animal cells in bioreactors the maintenance of a fixed pH-value is of elementary importance. In a perfused cell culture system the pH-value is measured by electrochemical flow rate sensors. For the acceptance of a technical solution the integration in complex workplaces is necessary, thereby small size and low costs are the main features. For that a special measurement module was developed, consisting of measuring amplifier and microcontroller component with CAN-Bus-interface.

[Development of a CAN (controller area network) bus for modules of a perfused cell culture system]. / Entwicklung einer CAN-Bus-Kopplung für Module eines perfundierten Zellkultursystems.

For computer-aided data acquisition and automated running of experiments in cell cultivation reactors networking of all sensoric and actoric devices is required. A low cost and high performance solution to this demand can be found by using the widely established CAN (Controller Area Network) bus. If standard PCs running MS Windows are used to control this network special measures have to be taken to prevent loss of data caused by differences in the computing power of the PC on one hand and microcontroller-based devices on the other.

[Recording and control of culture parameters in perfused cell culture systems]. / Erfassung und Steuerung von Versorgungsparametern in perfundierten Zellkultursystemen.

For the cultivation of animal cells perfused cell culture systems are advantageous. The measurement of the process parameters for the check of the culture is difficult through the small volumes and dimensions. A developed measuring system allows the measuring of the parameters temperature, pH-value and dissolved oxygen.

Chloride channel antagonists perturb growth and morphology of Neurospora crassa.

The chloride channel antagonists anthracene-9-carboxylic acid, ethacrynic acid and niflumic acid were found to be fungistatic and morphogenic when tested against the ascomycete Neurospora crassa. Potency increased with decreasing pH, suggesting that the protonated forms of the compounds were active. Niflumic acid produced the most pronounced growth aberrations which may reflect an ability to acidify the cytoplasm and block the plasma membrane anion channel of N. crassa.

CD28-mediated induction of proliferation in resting T cells in vitro and in vivo without engagement of the T cell receptor: evidence for functionally distinct forms of CD28.

JJ316 and JJ319 are rat CD28-specific monoclonal antibodies (mAb) of the gamma1 kappa isotype with identical co-stimulatory potency. At a concentration 100-1000-fold higher than that required for co-stimulation, JJ316, but not JJ319 induces massive proliferation of all T cell subsets in vitro without T cell receptor (TCR) triggering. "Direct" stimulation by JJ316 is fully blocked by JJ319, indicating that it is not due to cross-reactivity of JJ316 with the TCR complex or other activating receptors. JJ316 binds much more slowly to primary T cells than JJ319, whereas both antibodies bind with similar kinetics to CD28-transfected L-929 cells, suggesting that JJ316 binding to T cells requires redistribution or a conformational change of CD28. In vivo, JJ316 but not JJ319 induces rapid and transient proliferation of most CD4 T cells and, indirectly, of B cells. These data show that TCR engagement is not an absolute prerequisite either in vitro or in vivo for the induction of T cell proliferation through CD28 and suggest that mAb JJ316 is able to stimulate resting T cells directly by recruiting CD28 molecules from an inactive to an active form.