Inherited anxiety-related parent-infant dyads alter LHPA activity.

The pathogenesis of post-traumatic stress disorder (PTSD) is incompletely understood. We hypothesize that disruptions in mother-child relations may be a key contributor to development of PTSD. A normal and healthy separation-individuation process requires adaptations of self- and interactive contingency in both the mother and her child, especially in early childhood development. Anxious mothers are prone to overprotection, which may hinder the individuation process in their children. We examined long-term stress hormones and other stress markers in subjects three generations removed from the Holocaust, to assess the long-term consequences of inherited behavioral and physiological responses to prior stress and trauma. Jewish subjects who recalled overprotective parental behavior had higher hairsteroid-concentrations and dampened limbic-hypothalamic-pituitary-adrenal (LHPA) axis reactivity compared to German and Russian-German subjects with overprotective parents. We suggest that altered LHPA axis activity in maternally overprotected Jewish subjects may indicate a transmitted pathomechanism of "frustrated individuation" resulting from cross-generational anti-Semitic experiences. Thus measurements of hairsteroid-concentrations and parenting practices may have clinical value for diagnosis of PTSD. We propose that this apparent inherited adaptivity of LHPA axis activity could promote higher individual stress resistance, albeit with risk of an allostatic overload.

Vancomycin therapeutic drug monitoring: is there a consensus view? The results of a UK National External Quality Assessment Scheme (UK NEQAS) for Antibiotic Assays questionnaire.

This study investigated vancomycin therapeutic drug monitoring (TDM) and issues related to patient management. Questionnaires were distributed to 310 participants in the UK National External Quality Assessment Scheme (NEQAS) for Antibiotic Assays. The response rate was 57.4%. The majority (76%) had an 'in-house' assay service based, almost exclusively, in the microbiology department, and a fluorescence polarization immunoassay (FPIA) was used by 97%. Almost half (48.7%) had an assay service available for 24 h/day, 7 days/week and 92.7% expected same-day results. The majority (80%) had issued guidelines for vancomycin use. A 12 hourly initial dosing regimen was used by 89%. Trough assay samples were taken <10 min before the dose by 91.5%. For post-dose assay samples, 44% took a sample at 1 h, 28% at 2 h and the remainder at 'other' times. For trough target ranges, 93% quoted <10 mg/L or 5-10 mg/L. There was no consensus with regard to post-dose assay sample times and 23 ranges were quoted. The majority (74.4%) regarded a trough level of >or=10 mg/L as 'toxic' but 13 concentrations were quoted as toxic post-dose measurements. In conclusion, there was a wide variability and poor consensus with regard to post-dose vancomycin assay sampling times, target ranges and what constituted a toxic level.

A simple, isocratic high-performance liquid chromatography assay for linezolid in human serum.

A rapid high-performance liquid chromatography (HPLC) assay for the detection of linezolid in human serum was developed. The method used a Hypersil 5ODS stationary phase. The mobile phase was 1% ortho-phosphoric acid, 30% methanol, 2 g/L heptane sulphonic acid, pH 5. UV absorbance detection was used (lambda(max) 254 nm). Samples were prepared by mixing with acetonitrile and an injection volume of 20 microL was used. The inter- and intra-day assay reproducibility were assessed. Assay linearity, specificity and accuracy were investigated. The detection limit and recovery of linezolid from serum were determined. In addition, the stability of linezolid, stored under a variety of conditions, was assessed. The retention time of linezolid was c. 6.5 min. The intra- and inter-day reproducibility was good and the assay was linear across the therapeutic range. Serum recovery was c. 100% at all concentrations tested. The detection limit was 0.1 mg/L and the assay was accurate. The assay was specific as there was no significant interference with the linezolid peak. Linezolid was demonstrated to be stable. This rapid assay is ideal for busy clinical laboratories with basic HPLC equipment.

Antimicrobial activity of fluoroquinolone photodegradation products determined by parallel-line bioassay and high performance liquid chromatography.

The fluoroquinolones produce multiple photodegradation products. Little is known about these products, particularly whether any possess antimicrobial activity. To investigate this, we used the parallel-line bioassay to investigate discrepancies in zone of inhibition size in conjunction with high performance liquid chromatography (HPLC) analysis. A continuous flow photochemical reaction unit ('Beam-Boost') was used to partially photodegrade the fluoroquinolones ofloxacin, levofloxacin, ciprofloxacin and moxifloxacin (0.02 mM) by between 15 and 89%, as confirmed by HPLC. The concentration of residual parent fluoroquinolone in each irradiated sample was measured by HPLC and a non-irradiated control solution was prepared at the same concentration. These were compared by parallel-line bioassays using Escherichia coli, Enterobacter cloacae and Klebsiella oxytoca. With ofloxacin and levofloxacin, the zone size for the control solution was significantly less than that of the irradiated solutions, with >15% photodegradation in at least two of the indicator organisms, indicating that the photodegradation products possess antimicrobial activity. No difference was seen with ciprofloxacin at any level of photodegradation with any of the indicator organisms, nor with moxifloxacin at 30 and 54% photodegradation. A significant difference was observed with E. cloacae only, at 83% photodegradation.

UK NEQAS in antibiotic assays.

How providers of external quality assessment (EQA) programmes relate to and interact with the monitors and watchdog of clinical laboratory performance in the UK is described. With regard to the quality of antibiotic assays, the changes in methodologies and in performance quality between 1971 (when the UK NEQAS for Antibiotic Assays began) and 1999 is reviewed. How improvements in performance and changes of methodology are related is discussed. The findings and conclusions of two experimental pilot EQA distributions (the teicoplanin assay and serum bactericidal test) are also discussed.

Teicoplanin therapy for Staphylococcus aureus septicaemia: relationship between pre-dose serum concentrations and outcome.

Logistic regression analysis was performed on data drawn from a clinical trials database for Staphylococcus aureus septicaemia treated with teicoplanin. Variables analysed were age, body weight, mean pre-dose and post-dose serum teicoplanin concentrations, mean dose (mg or mg/kg body weight) and combination versus monotherapy. Only two variables correlated with clinical outcome at a significance level better than 0.05: age (P = 0.012) and mean pre-dose serum concentration (P = 0.010). The probability of successful treatment declined with age and increased with mean pre-dose serum concentration.

Difficulties in the assay of liposomal amikacin (MiKasome) in serum.

Antibiotic-free human serum was spiked with known concentrations of liposomal amikacin and assayed on the Abbott TDx System, using polarization fluoroimmuno assay (PFIA) kits from Abbott Laboratories, Oxis and Sigma. Although all three kits gave a linear response, the Abbott and Oxis kits showed very low recovery (<21%) with only the Sigma kit giving near 100% recovery. Heating samples at 56 degrees C for 30 min improved recovery with the Abbott and Oxis kits (75-80% of target value), but decreased recovery with the Sigma kit (85% of target value). The loss of amikacin from liposomal amikacin, as measured using the Sigma kit, was related to both temperature and duration of heating, reaching a maximal loss of 21% after 1 h at 60 degrees C.

Assays for therapeutic monitoring and pharmacokinetic investigations of aminoglycosides: quality aspects.

Clinical laboratory investigations used to aid antimicrobial chemotherapy of serious infection include routine sensitivity testing and, in the case of those drugs with a narrow therapeutic range, routine assays for therapeutic monitoring to assist with dosage individualization. Tests must be of a sufficiently high quality to be clinically useful. Laboratories ensure quality through standard operating procedures, internal quality control procedures, and participation in external quality assessment (EQA) programs. This article demonstrates how EQA returns to the United Kingdom National External Quality Assessment Scheme for Antibiotic Assays and the activity of the United Kingdom National Quality Assurance Advisory Panel showed marked improvement in the technical quality of assays as exemplified by gentamicin assays. The article also highlights additional quality concerns not subject to EQA.

The pharmacokinetics of intravenous ciprofloxacin 400 mg 12 hourly in patients with severe sepsis: the effect of renal function and intra-abdominal disease.

Serum concentrations of ciprofloxacin were reviewed in 22 patients given ciprofloxacin 400 mg intravenously 12 hourly for severe infection. No dosage modifications were made in patients with renal impairment. Patients who had either bowel or liver pathology in addition to renal failure had significantly higher serum concentrations than all other patients. Dosage reduction of ciprofloxacin in patients with severe sepsis and impaired renal function is not required unless they have co-existent intra-abdominal disease.

Half-life of intracameral gentamicin after phacoemulsification.

PURPOSE: To determine the half-life of intracameral gentamicin administered during phacoemulsification. SETTING: Southampton Eye Unit, Southampton General Hospital, England. METHODS: Thirty-one patients having scleral tunnel phacoemulsification were given intracameral gentamicin in the irrigation fluid. Samples of fluid were taken from the anterior chamber at the end of the operation and at various times postoperatively. The concentration of gentamicin in the samples was determined by fluorescence polarization immunoassay and the half-life calculated for a single compartment model using a peeling algorithm. RESULTS: The concentration of gentamicin in the anterior chamber after phacoemulsification decreased by half every 51 minutes (95% confidence interval, 42 to 66 minutes). CONCLUSION: Intracameral gentamicin was cleared from the anterior chamber after phacoemulsification at a rate that prevents the maintainance of the bactericidal levels required for reliable antibiotic prophylaxis.

The pharmacokinetics of once-daily oral 400 mg ofloxacin in patients with peritonitis complicating continuous ambulatory peritoneal dialysis.

Seven patients with end-stage renal disease requiring support by continuous ambulatory peritoneal dialysis received once-daily 400 mg oral ofloxacin for 7 days for the treatment of bacterial peritonitis. Serum and peritoneal dialysis fluid (PDF) were collected for assay throughout the course of the study and for 5 days thereafter. Ofloxacin, desmethyl ofloxacin and ofloxacin-N-oxide accumulated over the course of therapy and could still be detected in serum and PDF 5 days after the end of therapy. The mean elimination half-life of ofloxacin in serum was 32 +/- 7 h, desmethyl ofloxacin 45 +/- 26 h and for ofloxacin-N-oxide 44 +/- 15 h. The total mean recovery of ofloxacin and its metabolites from the PDF was 15.4%. This regimen results in serum and PDF concentrations likely to be effective for the treatment of infection for at least 10 days.

Evaluation of Innofluor fluorescence polarization immunoassay kits for the determination of serum concentrations of gentamicin, tobramycin, amikacin and vancomycin.lesassays@ukneqasaa.win-uk.net.

The Innofluor fluorescence polarization immunoassay (FPIA) kits for gentamicin, tobramycin, amikacin and vancomycin were evaluated on an Abbott TDX analyser. Intra-assay reproducibility was excellent with a coefficient of variation of <3% for all analytes. The coefficient of variation for inter-assay reproducibility was usually <5%. Assay linearity was good and standard curve stability was seen with kits of the same batch for at least 32 days. Using clinical samples, the results obtained with the Innofluor FPIA reagents correlated well with those obtained using Abbott FPIA reagents, but Innofluor gentamicin and amikacin results were slightly higher than Abbott results (P < 0.001). Results of UK NEQAS returns showed acceptable accuracy for the Innofluor kits, but mean Innofluor gentamicin returns were 4% higher (P = 0.001) and mean vancomycin returns were 5% lower (P = 0.001) than overall mean returns. Innofluor and Abbott vancomycin assay reagents showed similar cross-reactivity to degraded vancomycin.

The elution and binding characteristics of rifampicin for three commercially available protein-sealed vascular grafts.

Rifampicin was absorbed onto gelatin-sealed (Gelsoft and Unigraft) or collagen-sealed (Hemashield) vascular grafts by soaking for 15 min in a 1000 mg/L solution. Bound drug was then eluted from the grafts by incubation in phosphate buffered saline (PBS) at 37 degrees C and at timed intervals the concentration of rifampicin remaining in the grafts was determined. Although all three grafts contained high concentrations of rifampicin immediately after absorption of drug, rifampicin concentrations rapidly fell during elution with PBS to approximately 1.25 mg/kg of graft after 5 h incubation in PBS, indicating that most of the rifampicin absorbed to the grafts was only loosely bound. However, once this loosely bound fraction had been removed there was a much slower elution of the remaining rifampicin from the grafts, suggesting a second and much more tightly bound fraction. The tightly bound fraction eluted with an apparent half-life of 47-76 h, depending on the graft, and extrapolation back to time zero from this phase suggests that only a very small amount of the rifampicin is tightly bound to the graft after initial soaking (0.6-1.3 mg/kg).

The quality of clinical serum teicoplanin assays: an experimental European EQA distribution.

Two sets of six samples of human serum spiked with predetermined concentrations of teicoplanin were circulated, 6 months apart, to 22 European laboratories for the purposes of external quality assessment (EQA). The laboratories returned assay results using either bioassay, high-performance liquid chromatography (HPLC) or fluorescence polarization immunoassay (FPIA). FPIA was the most popular method and performance was generally satisfactory. Some laboratories using HPLC or microbiological assay performed satisfactorily but others did not. Only seven laboratories (32%) showed consistently satisfactory performance. There appears to be a need for continuing EQA of clinical teicoplanin assays.

Pharmacokinetics of oral and intravenous ofloxacin in children with multidrug-resistant typhoid fever.

The pharmacokinetics of oral and intravenous ofloxacin (7.5 mg.kg of body weight-1 given over 30 min) were studied in an open crossover study of 17 Vietnamese children, aged between 5 and 14 years, with acute uncomplicated typhoid fever. Following oral administration, the median (95% confidence interval [CI]) time to peak concentration of ofloxacin in serum (Cmax) was 1.7 h (1.4 to 1.9 h) and the mean (95% CI) Cmax was 5.5 mg.liter-1 (4.7 to 6.3 mg.liter-1) compared with a Cmax of 8.7 mg.liter-1 (7.6 to 9.7 mg.liter-1) following the intravenous infusion. The median (95% CI) total apparent volume of distribution following the first intravenous dose, 1.35 liter.kg-1 (1.17 to 1.73 liter.kg-1), was significantly larger than that following the second dose, 0.99 liter.kg-1 (0.86 to 1.17 liter.kg-1; P < 0.0005), although the estimates for systemic clearance were similar: 0.255 liter.kg-1 h-1 (0.147 to 0.325 liter.kg-1 h-1) compared with 0.172 liter.kg-1 h-1 (0.127 to 0.292 liter.kg-1 h-1; P = 0.14). The mean residence times (95% CI) following intravenous and oral administration were similar: 5.24 h (4.84 to 6.58 h) and 6.24 h (5.32 to 7.85 h), respectively. The mean (95% CI) oral bioavailability was 91% (74 to 109%). The peak concentrations in serum were 10 to 100 times higher than the maximum MICs for ofloxacin against multidrug-resistant Salmonella typhi isolated in this area. Although the systemic clearance values were higher than those reported previously for adults, these data overall suggest that weight-or area-adjusted dose regimens for the treatment of typhoid in older children should be the same as those for adults.