Evaluation of loop-mediated isothermal amplification for the rapid identification of bacteria and resistance determinants in positive blood cultures.

The use of molecular assays to rapidly identify pathogens and resistance genes directly from positive blood cultures (BCs) contribute to shortening the time required for the diagnosis of bloodstream infections. In this work, loop-mediated isothermal amplification (LAMP) assays have been examined for their potential use in BC diagnosis. Three different assays were applied. The commercially available eazyplex® MRSA test detects Staphylococcus aureus, S. epidermidis, mecA, and mecC. Two in-house assays [Gram-positive (GP) and Gram-negative (GN)] have been developed for the detection of streptococci, enterococci, vanA, vanB, Pseudomonas spp., Enterobacteriaceae, and the bla CTX-M family. A total of 370 positive BCs were analyzed. LAMP test results were obtained within 30 min, including sample preparation. Amplification was measured by real-time fluorescence detection. The threshold time for fluorescence intensity values ranged from 6.25 to 13.75 min. The specificity and sensitivity of the assays varied depending on the target. Overall, from 87.7% of BCs, true-positive results were obtained, compared to routine standard diagnosis. Twenty-one tests were true-negative because of the lack of an appropriate target (5.7%). The concordance of positive test results for resistance genes with subsequent antibiotic susceptibility testing was 100%. From 15 BC bottles with mixed cultures, eazyplex® assays produced correct results in 73% of the cases. This study shows that LAMP assays are fast and cost-saving tools for rapid BC testing in order to expedite the diagnostic report and improve the antibiotic stewardship for sepsis patients.

Gentamicin coating of plasma chemical oxidized titanium alloy prevents implant-related osteomyelitis in rats.

Implant related infection is one of the most feared and devastating complication associated with the use of orthopaedic implant devices. Development of anti-infective surfaces is the main strategy to prevent implant contamination, biofilm formation and implant related osteomyelitis. A second concern in orthopaedics is insufficient osseointegration of uncemented implant devices. Recently, we reported on a macroporous titanium-oxide surface (bioactive TiOB) which increases osseointegration and implant fixation. To combine enhanced osseointegration and antibacterial function, the TiOB surfaces were, in addition, modified with a gentamicin coating. A rat osteomyelitis model with bilateral placement of titanium alloy implants was employed to analyse the prophylactic effect of gentamicin-sodiumdodecylsulfate (SDS) and gentamicin-tannic acid coatings in vivo. 20 rats were randomly assigned to four groups: (A) titanium alloy; PBS inoculum (negative control), (B) titanium alloy, Staphylococcus aureus inoculum (positive control), (C) bioactive TiOB with gentamicin-SDS and (D) bioactive TiOB plus gentamicin-tannic acid coating. Contamination of implants, bacterial load of bone powder and radiographic as well as histological signs of implant-related osteomyelitis were evaluated after four weeks. Gentamicin-SDS coating prevented implant contamination in 10 of 10 tibiae and gentamicin-tannic acid coating in 9 of 10 tibiae (infection prophylaxis rate 100% and 90% of cases, respectively). In Group (D) one implant showed colonisation of bacteria (swab of entry point and roll-out test positive for S. aureus). The interobserver reliability showed no difference in the histologic and radiographic osteomyelitis scores. In both gentamicin coated groups, a significant reduction of the histological osteomyelitis score (geometric mean values: C = 0.111 ± 0.023; D = 0.056 ± 0.006) compared to the positive control group (B: 0.244 ± 0.015; p < 0.05) was observed. The radiographic osteomyelitis scores confirmed these histological findings.

Microbiological stability of solutions containing local anesthetics and opioids in closed infusion systems used for epidural analgesia.

Nine solutions containing opiod analgesics and local anesthetics as typically use in epidural catheters were tested for antimicrobial stability. Administration via a pefusor syringe requires several refill processes. It was shown that repetitive refilling of the syringes did not result in any microbiological contamination.

Oral delivery of macromolecules: rationale underpinning Gastrointestinal Permeation Enhancement Technology (GIPET).

Oral delivery of macromolecular drugs, particularly peptides and proteins, is the focus of many academic and industrial laboratories. Armed with an increased understanding of the structure and regulation of intestinal epithelial junctional complexes of the paracellular barrier, the development of permeation enhancement technology initially focused on the specific and reversible opening of tight junctions in order to enable oral delivery. Despite intense research, none of these specific tight junction-opening technologies has yet been approved in an oral drug product, likely because of poor efficacy. Less specific enhancer technologies with a long history of safe use in man have additional surfactant-like effects on the transcellular pathway that lead to improved efficacy. These are likely to be the first to market for selected poorly permeable peptides. This review presents a summary of some approaches taken to intestinal permeation enhancement and explores in detail the oral delivery system developed by Merrion Pharmaceuticals, Gastrointestinal Permeation Enhancement Technology (GIPET).

[Danger of infection from communion cups--an underestimated risk?]. / Infektionsgefährdung durch Abendmahlskelche--eine unterschätzte Gefahr?

The problem of a risk of infection from the common use of chalices has been discussed controversially in literature. Opinions were mainly based on laboratory experiments and theoretical considerations. The authors examined bacterial counts and species existing under normal conditions after communion. For this purpose, contact samples were taken from the inside and outside of chalices at the rim. Staphylococci and alpha-haemolytic streptococci were found on all chalices examined. On more than 80%, there were apathogenic micrococci, nonhaemolytic streptococci, apathogenic neisseria and apathogenic corynebacteria as well as lactobacilli and bacilli. Staphylococcus aureus was found on 26.4% of chalices. Although the risk of infection for healthy persons from a commonly used chalice can be rated as low, it should not be underestimated for persons with reduced resistance and immunocompetence, or with reduced defences as a result of therapeutic measures. From the hygienic point of view, the most favourable approaches to avoid infection would be the use of individual chalices for all participants in the communion or the immersion of wafers or bread in wine or in grape juice by the priest (intinction).

The value of a postoperative radiation therapy in FIGO stage I and II endometrial cancers.

PURPOSE: The incidence of endometrial cancer has increased considerably within the last decades. Due to early symptoms more than 88% of all patients present initially with FIGO stage I or II disease. PATIENTS AND METHODS: One hundred and forty-four patients out of 192 patients with stage I endometrial cancers received surgical treatment alone. Like the remaining 48 patients 39 out of 65 patients with stage II tumors received adjuvant radiation therapy with 50 Gy to the true pelvis. RESULTS: Independent on the chosen treatment results in stage I were influenced by the depth of myometrial invasion (p = 0.023). With a crude 5-year survival rate of 88% patients with less than 33% reached life expectation of matched healthy women. With deeper infiltration crude survival rates were as low as 77%. In stage II tumors 5-year crude survival was elevated 60 to 80% independent on the depth of myometrial infiltration by adjuvant radiation therapy (p = 0.11). CONCLUSIONS: Despite its potential curability and the general knowledge that the prognosis of endometrial carcinomas is governed by a number of factors there are no generally accepted standard treatment modalities available Thus there is great demand for randomized clinical trials.

[Effect of phosphate and inhibitors of protein synthesis on the growth and tentoxin production of Alternaria alternata (Fr.) Keissler]. / Einfluss von Phosphat und Inhibitoren der Proteinsynthese auf das Wachstum und die Tentoxinbildung von Alternaria alternata (Fr.) Keissler.

The synthesis of a phytotoxic substance, tentoxin, by Alternaria alternata (Fr.) Keissler is negatively controlled by inorganic phosphate. We show here that the influence of phosphate on the production of this secondary metabolite is time-dependent. By using inhibitors of protein synthesis (p-fluorphenylalanine, cycloheximide) together with a resuspension technique, attempts were made to find out when the enzymes responsible for tentoxin synthesis are formed.

Nonisothermal kinetics with programmed temperature steps.

Data required for predicting the stability of an active principle in solution can be obtained by two kinetic methods. With the isothermal method, the degradation rate constants are determined at different temperatures, which are kept constant throughout the experiment. With the nonisothermal method, the temperature is increased with time. This paper describes a nonisothermal kinetic method in which the temperature is increased in consecutive equal steps. The results are compared with those obtained by the conventional isothermal method. The values for the activation energy are approximately the same by both methods. Although the technique of nonisothermal kinetics demands sophisticated equipment and high experimental accuracy, it provides a continuous picture over a wide temperature range.