Agreement of different methods for assessment of progesterone concentrations in beef cattle serum.

Progesterone (P4) is responsible for the main reproduction processes. Concentration of P4 varies widely among different determination methods, and interpretation of these values may be difficult. The objective of the current study was to assess the agreement of three different enzyme immunoassays (ELISA) in relation to radioimmunoassay (RIA) of P4 concentration assessment of beef cow serum samples. Samples were collected randomly considering high (pregnant cows) and low (non-pregnant cows) P4 concentrations. Depending on the P4 assessment method, four groups were created as follows: Group 1 - direct samples assessed by ELISA, Group 2 - extracted samples assessed by ELISA, Group 3 - samples assessed by automated ELISA, and Group 4 - samples assessed by RIA. The mean progesterone concentration was 4.50 ng/mL, 1.24 ng/mL, 4.07 ng/mL and 4.39 ng/mL from Group 1 to Group 4, respectively. The mean difference (MD) between Group 1, Group 2 and Group 3 individually compared with Group 4 was -0.10 ± 1.24 ng/mL, 3.15 ± 3.58 ng/mL and 0.33 ± 1.42 ng/mL, and the 95% confidence interval (CI) for the differences (s) was from -0.99 to 0.78 ng/mL, from 0.59 to 5.71 ng/mL, and from -0.69 to 1.34 ng/mL, respectively. The confidence interval for the lower and upper limit of the agreement ranged from -4.12 to -1.05 ng/mL and from 0.84 to 3.91 ng/mL between Group 1 and Group 4, from -8.45 to 0.42 ng/ mL and from 5.88 to 14.75 ng/mL between Group 2 and Group 4, from -4.29 to -0.76 ng/mL, and from 1.41 to 4.94 ng/mL between Group 3 and Group 4. Our findings show that the best agreement with RIA was observed for Group 1 and Group 3, while the agreement in the extraction method was least accurate.

Vancomycin elution from a biphasic ceramic bone substitute.

OBJECTIVES: The aim of this study was to analyze drain fluid, blood, and urine simultaneously to follow the long-term release of vancomycin from a biphasic ceramic carrier in major hip surgery. Our hypothesis was that there would be high local vancomycin concentrations during the first week with safe low systemic trough levels and a complete antibiotic release during the first month. METHODS: Nine patients (six female, three male; mean age 75.3 years (sd 12.3; 44 to 84)) with trochanteric hip fractures had internal fixations. An injectable ceramic bone substitute, with hydroxyapatite in a calcium sulphate matrix, containing 66 mg of vancomycin per millilitre, was inserted to augment the fixation. The vancomycin elution was followed by simultaneously collecting drain fluid, blood, and urine. RESULTS: The antibiotic concentration in the drain reached a peak during the first six hours post-surgery (mean 966.1 mg/l), which decreased linearly to a mean value of 88.3 mg/l at 2.5 days. In the urine, the vancomycin concentration reached 99.8 mg/l during the first two days, followed by a logarithmic decrease over the next two weeks to reach 0 mg/l at 20 days. The systemic concentration of vancomycin measured in blood serum was low and decreased linearly from 2.17 mg/l at one hour post-surgery to 0 mg/l at four days postoperatively. CONCLUSION: This is the first long-term pharmacokinetic study that reports vancomycin release from a biphasic injectable ceramic bone substitute. The study shows initial high targeted local vancomycin levels, sustained and complete release at three weeks, and systemic concentrations well below toxic levels. The plain ceramic bone substitute has been proven to regenerate bone but should also be useful in preventing bone infection.Cite this article: M. Stravinskas, M. Nilsson, A. Vitkauskiene, S. Tarasevicius, L. Lidgren. Vancomycin elution from a biphasic ceramic bone substitute. Bone Joint Res 2019;8:49-54. DOI: 10.1302/2046-3758.82.BJR-2018-0174.R2.

Factors associated with severity in invasive community-acquired Staphylococcus aureus infections in children: a prospective European multicentre study.

Staphylococcus aureus is the main pathogen responsible for bone and joint infections worldwide and is also capable of causing pneumonia and other invasive severe diseases. Panton-Valentine leukocidin (PVL) and methicillin-resistant S. aureus (MRSA) have been studied as factors related with severity in these infections. The aims of this study were to describe invasive community-acquired S. aureus (CA-SA) infections and to analyse factors related to severity of disease. Paediatric patients (aged 0-16 years) who had a CA-SA invasive infection were prospectively recruited from 13 centres in 7 European countries. Demographic, clinical and microbiological data were collected. Severe infection was defined as invasive infection leading to death or admission to intensive care due to haemodynamic instability or respiratory failure. A total of 152 children (88 boys) were included. The median age was 7.2 years (interquartile range, 1.3-11.9). Twenty-six (17%) of the 152 patients had a severe infection, including 3 deaths (2%). Prevalence of PVL-positive CA-SA infections was 18.6%, and 7.8% of the isolates were MRSA. The multivariate analysis identified pneumonia (adjusted odds ratio (aOR) 13.39 (95% confidence interval (CI) 4.11-43.56); p 0.008), leukopenia at admission (<3000/mm(3)) (aOR 18.3 (95% CI 1.3-259.9); p 0.03) and PVL-positive infections (aOR 4.69 (95% CI 1.39-15.81); p 0.01) as the only factors independently associated with severe outcome. There were no differences in MRSA prevalence between severe and nonsevere cases (aOR 4.30 (95% CI 0.68- 28.95); p 0.13). Our results show that in European children, PVL is associated with more severe infections, regardless of methicillin resistance.

Nosocomial dissemination of VIM-2-producing ST235 Pseudomonas aeruginosa in Lithuania.

Pseudomonas aeruginosa multidrug resistance, and particularly the production of carbapenemases linked to international high-risk clones, is of growing concern. While high levels of carbapenem resistance (>60 %) have been reported in Lithuania, so far, there is no information on the underlying mechanisms. Thus, the aim of this work was to determine the molecular epidemiology and prevalence of acquired carbapenemases among 73 carbapenem-resistant P. aeruginosa isolates recovered in a hospital from Kaunas, Lithuania in 2011-2012. The presence of acquired carbapenemases was evaluated through phenotypic (modified Hodge test, cloxacillin inhibition test, double-disc synergy test) and genetic methods [polymerase chain reaction (PCR) and sequencing]. Clonal relatedness was assessed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Acquired ß-lactamases were detected in 19 (26 %) of the isolates, whereas resistance was exclusively chromosomal (OprD inactivation ± AmpC hyperproduction) in the remaining 54 (74 %) isolates. The acquired ß-lactamases detected included 16 VIM-2, one PER-1 and two GES enzymes. PFGE revealed that 15 of the 16 VIM-2 isolates belonged to a single clone, identified as the international high-risk clone ST235 by MLST. bla VIM-2 was preceded by aacA7 in a class I integron, similar to epidemic ST235 isolates described in nearby countries. Additionally, sequencing of bla GES revealed the presence of the carbapenem-hydrolysing enzyme GES-5 in one of the isolates and a novel GES variant, designated GES-27, in the other. GES-27 differed from GES-5 by a single amino acid substitution, proline 167, that was replaced by glutamine. Increasing emergence and dissemination of concerning resistance mechanisms and international clones warrants global surveillance and control strategies.

Augmented renal clearance--an evolving risk factor to consider during the treatment with vancomycin.

WHAT IS KNOWN AND OBJECTIVE: Augmented renal clearance (ARC) is a new phenomenon in patients' pathophysiology without universally accepted aetiology and with various incidence rates most often described in critically ill patients in the Intensive Care Unit (ICU). The objective of this retrospective observational comparative study was to estimate the incidence rate of ARC in patients with different medical conditions employing steady state trough vancomycin serum concentrations (VSCss) for analysis. METHODS: All patients tested for VCSss during two years (2010-2011) in a tertiary level hospital were analysed and 77 VSCs were eligible for analysis: 38 (50%) and 39 cases were assigned to the ARC (eCrClCG (creatinine clearance, estimated by Cockcroft-Gault) > 130 mL/min) and the control groups (eCrClCG in the range 90-130 mL/min) respectively. RESULTS AND DISCUSSION: Patients' age, mechanical ventilation and haemodynamically unstable status had significant association with ARC occurrence (P < 0.05). Majority of ARC patients (11 patients (61 %)) were managed in non-ICU settings. ARC event showed statistically significant higher risk for under dosage (RR (relative risk for subtherapeutic VSCss), 1.84; 95% CI, 1.23\x962.74; P = 0.011), and the correlation between different thresholds (eCrClCG >130 mL/min, ≥140 mL/min and ≥150 mL/min) for ARC and VSCss allows to predict decrease of VSCss in case of eCrClCG ≥150 mL/min: every increase of eCrClCG by 40 mL/min predicts clinically relevant decrease of VSCss by 1 mmol/L (1.49 mg/mL). WHAT IS NEW AND CONCLUSION: ARC cases lead to the doubled risk of subtherapeutic VSC, and this phenomenon is a significant event in patients in any hospital department. Investigation of medical patients' status relevant to this phenomenon needs to be continued.

Pseudomonas aeruginosa strains from nosocomial pneumonia are more serum resistant than P. aeruginosa strains from noninfectious respiratory colonization processes.

BACKGROUND: Serum resistance is regarded as a major virulence factor of bacteria and is thought to be mediated by O side chains of the lipopolysaccharides (LPS). We investigated the serum-resistance properties and O serogroups of Pseudomonas aeruginosa strains isolated from intensive careunit (ICU) patients with pneumonia and from the respiratory tract of ICU patients without respiratory tract infections. MATERIALS AND METHODS: 171 P. aeruginosa strains were consecutively isolated from bronchoalveolar lavage fluid or transtracheal aspirates of ICU patients with monobacterial nosocomial pneumonia and 49 strains were isolated from the respiratory tract of ICU patients without respiratory tract infections. All strains were O serogrouped using Oantigen- specific sera for 14 O serogroups and tested for their sensitivity to the serum's bactericidal effect. RESULTS: Using two different analyses, the frequency of serum-sensitive isolates was significantly lower in strains from patients with pneumonia (56.1%; n = 96/171 and 22.8%, n = 39/171, respectively) than in strains from asymptomatically colonized patients (73.46%; 36/49 and 38.8%, n = 19/49, respectively) (p = 0.03; OR = 2.163; 95% CI = 1.072-4.368 and p = 0.0289; OR = 2.144; 95% CI = 1.089-4.368, respectively). O serogrouping revealed higher frequency of the serogroups A (11.9% and 16.3%, respectively), B (14.3% and 21%), E (26.5% and 24.6%), and I (28.6% and 28%) in both strain collections. The frequency of serum-sensitive strains (13/28 and 3/45, respectively) was significantly lower among strains expressing the A and B serogroups, than for all other serogroups (p < 0.05). CONCLUSION: Strains isolated from patients with pneumonia and strains possessing O-A or O-B serogroups appear to have greater pathogenic potential by virtue of their ability to resist serum-mediated killing. The linkage, however, between the O serogroups, serum resistance, and a strain's virulence remains unclear at this stage.