Blood group AB increases risk for surgical necrotizing enterocolitis and focal intestinal perforation in preterm infants with very low birth weight.

Necrotizing enterocolitis (NEC) and focal intestinal perforation (FIP) are two of the most common emergencies of the gastrointestinal tract in preterm infants with very low birth weight (VLBW, birth weight < 1500 g). Identification of risk factors among these children is crucial for earlier diagnosis and prompt intervention. In this study, we investigated a relationship between ABO blood groups and the risk for surgical NEC/FIP. We genotyped the ABO locus (rs8176746 and rs8176719) in VLBW infants enrolled in a prospective, population-based cohort study of the German Neonatal Network (GNN). Of the 10,257 VLBW infants, 441 (4.3%) had surgical NEC/FIP. In univariate analyses, the blood group AB was more prevalent in VLBW infants with surgical NEC/FIP compared to non-AB blood groups (OR 1.51, 95% CI 1.07-2.13, p = 0.017; absolute risk difference 2.01%, 95% CI 0.06-3.96%). The association between blood group AB and surgical NEC/FIP was observed in a multivariable logistic regression model (OR of 1.58, 95% CI 1.10-2.26, p = 0.013) as well. In summary, our study suggests that the risk of surgical NEC and FIP is higher in patients with blood group AB and lower in those having non-AB blood groups.

Development of phantom materials with independently adjustable CT- and MR-contrast at 0.35, 1.5 and 3 T.

Quality assurance in magnetic resonance (MR)-guided radiotherapy lacks anthropomorphic phantoms that represent tissue-equivalent imaging contrast in both computed tomography (CT) and MR imaging. In this study, we developed phantom materials with individually adjustable CT value as well as [Formula: see text]- and [Formula: see text]-relaxation times in MR imaging at three different magnetic field strengths. Additionally, their experimental stopping power ratio (SPR) for carbon ions was compared with predictions based on single- and dual-energy CT. Ni-DTPA doped agarose gels were used for individual adjustment of [Formula: see text] and [Formula: see text] at [Formula: see text] and 3.0 T. The CT value was varied by adding potassium chloride (KCl). By multiple linear regression, equations for the determination of agarose, Ni-DTPA and KCl concentrations for given [Formula: see text] [Formula: see text] and CT values were derived and employed to produce nine specific soft tissue samples. Experimental [Formula: see text] [Formula: see text] and CT values of these soft tissue samples were compared with predictions and additionally, carbon ion SPR obtained by range measurements were compared with predictions based on single- and dual-energy CT. The measured CT value, [Formula: see text] and [Formula: see text] of the produced soft tissue samples agreed very well with predictions based on the derived equations with mean deviations of less than [Formula: see text] While single-energy CT overestimates the measured SPR of the soft tissue samples, the dual-energy CT-based predictions showed a mean SPR deviation of only [Formula: see text] To conclude, anthropomorphic phantom materials with independently adjustable CT values as well as [Formula: see text] and [Formula: see text] relaxation times at three different magnetic field strengths were developed. The derived equations describe the material specific relaxation times and the CT value in dependence on agarose, Ni-DTPA and KCl concentrations as well as the chemical composition of the materials based on given [Formula: see text] and CT value. Dual-energy CT allows accurate prediction of the carbon ion range in these materials.

Methodology paper: a novel phantom setup for commissioning of scanned ion beam delivery and TPS.

BACKGROUND: Commissioning of treatment planning systems (TPS) and beam delivery for scanned light ion beams is an important quality assurance task. This requires measurement of large sets of high quality dosimetric data in anthropomorphic phantoms to benchmark the TPS and dose delivery under realistic conditions. METHOD: A novel measurement setup is described, which allows for an efficient collection of a large set of accurate dose data in complex phantom geometries. This setup allows dose measurements based on a set of 24 small volume ionization chambers calibrated in dose to water and mounted in a holder, which can be freely positioned in a water phantom with various phantoms mounted in front of the water tank. The phantoms can be scanned in a CT and a CT-based treatment planning can be performed for a direct benchmark of the dose calculation algorithm in various situations. RESULTS: The system has been used for acceptance testing in scanned light ion beam therapy at Heidelberg Ion Beam Therapy Center for scanned proton and carbon ion beams. It demonstrated to be useful to collect large amounts of high quality data for comparison with the TPS calculation using various phantom geometries. CONCLUSION: The setup is an efficient tool for commissioning and verification of treatment planning systems. It is especially suited for dynamic beam delivery, as many data points can be obtained during a single plan delivery, but can be adapted also for other dynamic therapies, like rotational IMRT.

Pharmacodynamic comparison of LY3023703, a novel microsomal prostaglandin e synthase 1 inhibitor, with celecoxib.

To assess the safety, tolerability, and pharmacology of LY3023703, a microsomal prostaglandin E synthase 1 (mPGES1) inhibitor, a multiple ascending dose study was conducted. Forty-eight subjects received LY3023703, celecoxib (400 mg), or placebo once daily for 28 days. Compared with placebo, LY3023703 inhibited ex vivo lipopolysaccharide-stimulated prostaglandin E2 (PGE2 ) synthesis 91% and 97% on days 1 and 28, respectively, after 30-mg dosing, comparable to celecoxib's effect (82% inhibition compared to placebo). Unlike celecoxib, which also inhibited prostacyclin synthesis by 44%, LY3023703 demonstrated a maximal increase in prostacyclin synthesis of 115%. Transient elevations of serum aminotransferase were observed in one subject after 30-mg LY3023703 dosing (10× upper limit of normal (ULN)), and one subject after 15-mg dosing (about 1.5× ULN). Results from this study suggest that mPGES1 inhibits inducible PGE synthesis without suppressing prostacyclin generation and presents a novel target for inflammatory pain.

A motorized solid-state phantom for patient-specific dose verification in ion beam radiotherapy.

For regular quality assurance and patient-specific dosimetric verification under non-horizontal gantry angles in ion beam radiotherapy, we developed and commissioned a motorized solid state phantom. The phantom is set up under the selected gantry angle and moves an array of 24 ionization chambers to the measurement position by means of three eccentrically-mounted cylinders. Hence, the phantom allows 3D dosimetry at oblique gantry angles. To achieve the high standards in dosimetry, the mechanical and dosimetric accuracy of the phantom was investigated and corrections for residual uncertainties were derived. Furthermore, the exact geometry as well as a coordinate transformation from cylindrical into Cartesian coordinates was determined. The developed phantom proved to be suitable for quality assurance and 3D-dose verifications for proton- and carbon ion treatment plans at oblique gantry angles. Comparing dose measurements with the new phantom under oblique gantry angles with those in a water phantom and horizontal beams, the dose deviations averaged over the 24 ionization chambers were within 1.5%. Integrating the phantom into the HIT treatment plan verification environment, allows the use of established workflow for verification measurements. Application of the phantom increases the safety of patient plan application at gantry beam lines.

Integration and evaluation of automated Monte Carlo simulations in the clinical practice of scanned proton and carbon ion beam therapy.

Monte Carlo (MC) simulations of beam interaction and transport in matter are increasingly considered as essential tools to support several aspects of radiation therapy. Despite the vast application of MC to photon therapy and scattered proton therapy, clinical experience in scanned ion beam therapy is still scarce. This is especially the case for ions heavier than protons, which pose additional issues like nuclear fragmentation and varying biological effectiveness. In this work, we present the evaluation of a dedicated framework which has been developed at the Heidelberg Ion Beam Therapy Center to provide automated FLUKA MC simulations of clinical patient treatments with scanned proton and carbon ion beams. Investigations on the number of transported primaries and the dimension of the geometry and scoring grids have been performed for a representative class of patient cases in order to provide recommendations on the simulation settings, showing that recommendations derived from the experience in proton therapy cannot be directly translated to the case of carbon ion beams. The MC results with the optimized settings have been compared to the calculations of the analytical treatment planning system (TPS), showing that regardless of the consistency of the two systems (in terms of beam model in water and range calculation in different materials) relevant differences can be found in dosimetric quantities and range, especially in the case of heterogeneous and deep seated treatment sites depending on the ion beam species and energies, homogeneity of the traversed tissue and size of the treated volume. The analysis of typical TPS speed-up approximations highlighted effects which deserve accurate treatment, in contrast to adequate beam model simplifications for scanned ion beam therapy. In terms of biological dose calculations, the investigation of the mixed field components in realistic anatomical situations confirmed the findings of previous groups so far reported only in homogenous water targets. This work can thus be useful to other centers commencing clinical experience in scanned ion beam therapy.

Significantly increased CD70 up regulation on TEL-AML positive B cell precursor acute lymphoblastic leukemia cells following CD40 stimulation.

BACKGROUND: TEL-AML the most common genetic alteration in childhood precursor B acute lymphoblastic leukemia (BCP-ALL) is associated with a favorable prognosis. PATIENTS AND METHOD: We studied the expression of nerve growth factor/tumor necrosis factor receptor (NGFR/TNFR)/ligand family members on 108 primary BCP-ALL samples by flow cytometry and compared both their baseline expression and CD40-induced modulation on TEL-AML positive and negative leukemia samples. RESULTS: Our findings demonstrate that TEL-AML positive patients exhibit a significantly higher percentage of CD40, CD27 and p75NTR positive blasts at diagnosis. This might well contribute to the improved relapse-free survival of these patients assessed in Kaplan Meier analysis as CD27 and p75NTR directly mediate apoptotic signals. Furthermore CD40 ligation enhances antigen presenting and T cell stimulatory capacity via significant up regulation of CD70 while adequate response to physiological maturation signals as indicated by concomitant down regulation of CD27 is retained in TEL-AML positive leukemia. CONCLUSION: These data provide novel insights in immunological control mechanisms preserved in this leukemia subtype and suggest that not only treatment with chemicals such as HDAC inhibitors but also retained in vivo response to CD40 ligation contributes to improved immune surveillance in these patients which may add to a superior relapse-free survival observed particularly in the presence of other risk factors.

Exercise DVD effect on musculoskeletal disorders in professional orchestral musicians.

BACKGROUND: Professional musicians report a high prevalence of performance-related musculoskeletal disorders (PRMDs). Excessive muscle tension and fatigue have been reported as important factors contributing to PRMDs. AIMS: To evaluate feasibility and effectiveness of a specific exercise programme delivered via a digital video disc (DVD) targeting PRMDs and associated risk factors. METHODS: Volunteers from eight Australian symphony orchestras undertook two or more sessions per week over 12 weeks. Questionnaires were administered pre- and post-intervention with items including the frequency and severity of PRMDs, perceived exertion during different playing situations, per formance effects of the DVD and satisfaction rates. Musicians who had also participated in an equivalent face-to-face programme prior to this DVD trial compared the two interventions. RESULTS: One hundred and forty-four out of 576 musicians volunteered (25% uptake), and 50 participants completed a mean 2.1 (SD 0.42) sessions over the 12 week period (41% compliance). PRMD frequency and severity were significantly reduced post-intervention (P < 0.01). Participants reported benefits of the DVD on strengthening muscles, increasing ease of movement and improving flexibility related to playing. Despite this, perceived exertion levels during private practice, rehearsal and performance remained the same (not significant). Seventy-eight per cent of participants scored their overall experience of the use of the DVD as good or excellent. Owing to its convenience and detailed exercise demonstrations, the DVD was rated as better or much better overall than the face-to-face classes by 55% of participants who had experienced both. CONCLUSIONS: An exercise DVD was well received and appeared to be effective, convenient and safe in managing occupational-specific musculoskeletal disorders in musicians.

Prevalence of velopharyngeal insufficiency in woodwind and brass students.

BACKGROUND: Velopharyngeal insufficiency (VPI) is a disorder in which air leaks out through the nose, reducing performance quality and capacity in wind and brass players. There have been limited studies on the prevalence of this potentially career-threatening disorder. AIMS: To identify the prevalence of VPI in a sample of conservatorium level woodwind and brass student instrumentalists in Australia. METHODS: Wind and brass students from four music institutions were recruited by email invitation to complete an online survey. Results from 77 musicians were analysed for their knowledge and experience of VPI. Musicians who had experienced VPI provided information on the characteristics, symptoms and treatment or advice sought for the disorder. RESULTS: Of the 77 musicians included in the analysis, 44% had heard of VPI, 30% were aware of other musicians who had experienced VPI and 39% had personally experienced VPI. CONCLUSIONS: The results suggest that VPI may be a common occurrence in wind and brass players. Informal discussions with colleagues and music teachers also suggest that VPI is a frequent phenomenon. This group of musicians represents the largest sample surveyed about VPI to date.

Impact of bispectral index monitoring on postoperative delirium in patients undergoing aortic surgery.

BACKGROUND: Bispectral index monitoring can facilitate anesthesia care. We evaluated the association of Bispectral index with postoperative neurological outcome and delirium in patients undergoing aortic surgery. METHODS: From 2006 to 2009, 292 consecutive patients undergoing aortic surgery were retrospectively reviewed. Patients were classified into 5 groups according to Bispectral index reduction: Group I (≤15%), Group II (15-20%), Group III (20-25%), Group IV (25-30%), and Group V (>30%). RESULTS: The number of patients in each group was : 52 (17.8%), Group I; 125 (42.8%), Group II;68 (23.3%), Group III; 33 (11.3%), Group IV; 14 (4.8%), Group V. The incidence of delirium and neurological events was higher in Group IV and Group V(90.9% and 18.2% in Group IV, and 71% and 79% in Group V; both p<0.001). Only Group V showed a longer intensive care unit stay compared to Group I (13.5±10.3 vs 5.4±6.6 days; p=0.002), Group II (7.3±8.6 days, p=0.005) and Group III (6.7±6.5 days, p=0.015). Group V also showed a longer intubation time compared to Group I (228±211 vs 73±112 hours; p=0.008) and Group II (105±177 hours, p=0.002). CONCLUSIONS: Our data suggest a higher incidence of neurological deficits in patients with a Bispectral index reduction of >25% from baseline. Explanations for these findings are speculative with regard to the underlying mechanisms, and larger studies are warranted to clarify these issues.

Reduced expression and defective modulation of TNF receptor/ligand family molecules on proB-ALL blasts.

BACKGROUND: There is a subgroup of pediatric patients with an immature immunophenotype of proB-ALL that still poses a therapeutic challenge, even if the overall prognosis in B cell precursor acute lymphoblasic leukemia (BCP-ALL) is very good. Due to impaired treatment response these patients are prone to suffer relapse and are thus by definition stratified into the clinically defined high risk group receiving intensified chemotherapy. Besides response to chemotherapy long term prognosis is also influenced by immunological control mechanisms. Thus, high expression of the TNF receptor CD40 has been shown to prevent particularly late relapse in BCP-ALL suggesting a pivotal role of this regulatory molecule for maintenance of the remission status. PATIENTS AND METHODS: We therefore determined the baseline expression and CD40-mediated modulation of TNF receptor and costimulatory molecules in 5 patients with proB-ALL, 8 with preB-ALL and 22 with c-ALL performing FACS analysis. We particularly compared the TNF receptor status on proB-ALL blasts to the expression on more mature preB- and c-ALL blasts. RESULTS: Here, we demonstrate for the first time a significantly lower baseline expression and CD40-induced modulation capacity of TNF receptor and costimulatory molecules in pediatric proB-ALL compared to more mature precursor B-ALL blasts. CONCLUSION: The lower expression and defective capacity of proB-ALL blasts to respond to CD40 ligand stimulation might resemble the immature feature of these blasts and besides increased chemoresistance contribute to the impaired prognosis of these patients due to escape from apoptosis and immunological control mechanisms.

Collagen biosynthesis of mechanically loaded articular cartilage explants.

OBJECTIVE: The purpose of this study was to investigate systematically the effect of load amplitudes, frequencies and load durations of intermittently applied mechanical pressure on the biosynthesis of collagen and non-collagenous proteins (NCP) as well as on the water content of cultured bovine articular cartilage explants. METHODS: Cyclic compressive pressure was applied using a sinusoidal waveform of 0.5 Hz frequency with a peak stress of 0.1, 0.5 or 1.0 MPa for a period of 10s followed by a load-free period of 10, 100 or 1000s. These intermittent loading protocols were repeated for a total duration of 1, 3 or 6 days. During the final 18 h of experiments, the incorporation of [(3)H]-proline into collagen and NCP, the content of water as well as the deformation of loaded explants were determined. RESULTS: Intermittently applied, cyclic mechanical loading of articular cartilage explants consistently reduced the relative rate of collagen synthesis compared to load-free conditions. This reduced proportion of newly synthesized collagen among newly made proteins was independent of the mechanical stimuli applied. The release of newly synthesized collagen and NCP from loaded explants into the nutrient media was unaffected by any of the loading protocols applied. In addition, quantitative data are provided showing that only high amplitudes of loads and frequencies enhanced the water content of the explants. CONCLUSIONS: Previous studies reporting that osteoarthritic cartilage in vivo can synthesize elevated amounts of collagen imply that the loading protocols chosen were inadequate for simulating in vitro osteoarthritic-like alterations of collagen synthesis. In our experiments the collagen biosynthesis of chondrocytes was only minor responsive to alterations in mechanical stimuli, applied over a wide range. Thus, our results imply that the synthesis of these structural macromolecules is under the strict control of normal chondrocytes enabling them to maintain the shape of this physical demanded tissue.

Data-driven analysis approach for biomarker discovery using molecular-profiling technologies.

High-throughput molecular-profiling technologies provide rapid, efficient and systematic approaches to search for biomarkers. Supervised learning algorithms are naturally suited to analyse a large amount of data generated using these technologies in biomarker discovery efforts. The study demonstrates with two examples a data-driven analysis approach to analysis of large complicated datasets collected in high-throughput technologies in the context of biomarker discovery. The approach consists of two analytic steps: an initial unsupervised analysis to obtain accurate knowledge about sample clustering, followed by a second supervised analysis to identify a small set of putative biomarkers for further experimental characterization. By comparing the most widely applied clustering algorithms using a leukaemia DNA microarray dataset, it was established that principal component analysis-assisted projections of samples from a high-dimensional molecular feature space into a few low dimensional subspaces provides a more effective and accurate way to explore visually and identify data structures that confirm intended experimental effects based on expected group membership. A supervised analysis method, shrunken centroid algorithm, was chosen to take knowledge of sample clustering gained or confirmed by the first step of the analysis to identify a small set of molecules as candidate biomarkers for further experimentation. The approach was applied to two molecular-profiling studies. In the first study, PCA-assisted analysis of DNA microarray data revealed that discrete data structures exist in rat liver gene expression and correlated with blood clinical chemistry and liver pathological damage in response to a chemical toxicant diethylhexylphthalate, a peroxisome-proliferator-activator receptor agonist. Sixteen genes were then identified by shrunken centroid algorithm as the best candidate biomarkers for liver damage. Functional annotations of these genes revealed roles in acute phase response, lipid and fatty acid metabolism and they are functionally relevant to the observed toxicities. In the second study, 26 urine ions identified from a GC/MS spectrum, two of which were glucose fragment ions included as positive controls, showed robust changes with the development of diabetes in Zucker diabetic fatty rats. Further experiments are needed to define their chemical identities and establish functional relevancy to disease development.

[Assessment of the tetanus immune status in plasma donors of the Blood Donor Service of the Bavarian Red Cross]. / Evaluierung des Tetanus-Immunstatus bei Plasmaspendern des Blutspendedienstes des Bayerischen Roten Kreuz.

BACKGROUND AND OBJECTIVE: Tetanus is nowadays largely under control, as a result of the preventive immunization programme. This prophylactic measure is important also because there is no specific treatment. In Germany tetanus occurs almost exclusively in elderly, not adequately immunized persons. This study determined the existing immune status of plasma donors in the region of Ingolstadt, Bavaria. SUBJECTS AND METHODS: Tetanus IgG antibody concentrations were measured with the ELISA test in serum of 1742 plasma donors (891 women, 851 men, age range 18-64 years). RESULTS: An adequately protective immunity against tetanus was found in 98.7% of men and 94.1% of women, while 5 women and 3 men had no protection. Demographic data indicated that the immunization protected males longer than females, regardless of age, especially in those over 50 years of age (85% of females, 99% of males). CONCLUSION: Adequate tetanus protection was achieved in the overwhelming majority of test subjects. This demonstrates that the prevalence of protection is lower in females than in males, a difference most marked in those over 50 years of age. It is important that this age group in particular should, as part of health care, be informed by their medical practitioner of the dangers of tetanus and the advantages of immunization.

Characterization of CD34+ progenitor-derived dendritic cells pulsed with tumor cell lysate for a vaccination strategy in children with malignant solid tumors and a poor prognosis.

BACKGROUND: Children and adolescents with primary multifocal, refractory or relapsed malignant extracranial solid tumors still have a poor prognosis inspite of intensive standard radio-/chemotherapy. Here complementary immunomodulatory treatment modalities may prove beneficial as consolidation therapy following cytoreduction. Neuroblastoma, Ewing tumor and soft tissue sarcoma cells have principally been shown to be susceptible towards both cytotoxic and humoral effector mechanisms. Yet in vivo they are not capable of inducing an effective antitumor response which has been attributed to low level MHC expression and lack of costimulatory surface molecules. Professional antigen - presenting cells such as dendritic cells (DCs) in contrast are capable of activating unprimed T cells and are therefore ideal tools for vaccine generation. RESULTS: Here we demonstrate that DCs may be generated from CD34+ progenitor cells to clinical scale in a three to four week cell culture process including an initial expansion and subsequent differentiation and maturation steps. DCs derived from CD34+ progenitors express the expected marker profile and are highly effective in stimulating allogeneic T cell effectors. We also demonstrate that they effectively take up fluorescence-labelled tumor cell lysate. DISCUSSION: Having established a cell culture process for clinical scale DC production utilizing CD34+ progenitors as the cellular source we discuss the role of CD34+ derived DCs in clinical vaccination protocols. The rationale for a phase I/II DC dose escalation study for high risk pediatric patients with extracranial solid tumors assessing safety, immunological and clinical efficacy of serial combined intranodal and subcutaneous injections of tumor cell lysate-pulsed autologous DCs is delineated.

[Open versus arthroscopic shoulder stabilization. An analysis of the treatment costs]. / Offene vs. arthroskopische anteriore Schulterstabilisierung. Eine Kostenvergleichsanalyse.

In this study the total costs of clinical open and arthroscopic anterior shoulder stabilization were evaluated, analyzed and compared. From 1988 to 1998 147 patients underwent open (Bankart) or arthroscopic (ASK) anterior shoulder stabilization. We randomized two groups of 30 patients for each method (Bankart: 25 male, 5 female, 29 years of age; ASK: 25 male, 5 female, 26 years of age) and evaluated the costs of their clinical treatment. The total cost was significantly ( p<0.05, Mann-Whitney U-Test) higher for the open (5639 euro) than for the arthroscopic (4601 euro) therapy. There was a significant difference between the groups for the average cost of surgery (Bankart: 2741 euro; ASK: 2315 euro, p<0.05) and the average postoperative treatment cost (Bankart: 2202 euro; ASK: 1630 euro, p<0.05) whereas the average preoperative treatment cost was not significantly different (Bankart: 669 euro, ASK: 657 euro). The staff costs for the surgical procedure (Bankart: 1800 euro (32%), ASK: 1319 euro (29%)) and the postoperative staff costs of the nurses (Bankart: 1271 euro (23%), ASK: 997 euro (22%)) represented the greatest parts of the total costs. The average duration of the clinical treatment was 15.8 days for the open- and 12,4 days for the arthroscopic group.

Collection, storage, and filtration of in vivo study samples using 96-well filter plates to facilitate automated sample preparation and LC/MS/MS analysis.

The benefits of high-throughput bioanalysis within the pharmaceutical industry are well established. One of the most significant bottlenecks in bioanalysis is transferring in vivo-generated study samples from their collection tubes during sample preparation and extraction. In most cases, the plasma samples must be stored frozen prior to analysis, and the freeze/thaw (F/T) process introduces thrombin clots that are capable of plugging pipets and automated liquid-transfer systems. A new approach to dealing with this problem involves the use of Ansys Captiva 96-well 20-microm polypropylene filter plates to collect, store frozen, and filter plasma samples prior to bioanalysis. The samples are collected from the test subjects, and the corresponding plasma samples are placed directly into the wells of the filter plate. Two Duoseal (patent pending) covers are used to seal the top and bottom of the plate, and the plate is stored at down to -70 degrees C. Prior to sample analysis, the seals are removed and the plate is placed in a 96-well SPE manifold. As the plasma thaws, it passes (by gravity or mild vacuum) through the polypropylene filter into a 96-well collection plate. A multichannel pipet or automated liquid-transfer system is used to transfer sample aliquots without fear of plugging. A significant advantage of this approach is that, unlike other methods, issues related to incomplete pipetting are virtually eliminated. The entire process is rapid since thawing and filtering take place simultaneously, and if a second F/T cycle is required for reanalysis, it is not necessary to refilter the samples (additional clotting was not observed after three F/T cycles). This technique was tested using monkey, rat, and dog plasma and sodium heparin and EDTA anticoagulants. To assess the possibility of nonspecific binding to the polypropylene filter, a variety of drug candidates from diverse drug classes were studied. Validation data generated for two Lilly compounds from distinct classes, before and after filtering, are presented in this paper as practical examples of this technique. While LC/MS/MS is the primary method of bioanalysis in our laboratory, the technique presented in this paper is applicable to other forms of detection as well.

Determination of olanzapine in human blood by liquid chromatography-tandem mass spectrometry.

A liquid chromatographic-tandem mass spectrometric (LC-MS-MS) assay was developed and validated to quantitatively determine olanzapine (OLZ) concentrations in human blood. Liquid-liquid extraction, using n-butanol:cyclohexane (3:47, v/v), was used to isolate OLZ and its internal standard, LY170158, from the biological matrix. Chromatographic resolution of OLZ from endogenous interferences and known metabolites was accomplished with a MetaChem Monochrom HPLC column (4.6 x 150 mm, d(p) 5 microm). Detection occurred using a Perkin-Elmer Sciex API III Plus triple quadrupole mass spectrometer using positive ion APCI and multiple reaction monitoring (MRM). The linear dynamic range was from 5 to 500 ng ml(-1) based on a 0.25-ml aliquot of human blood. The inter-day precision (%RSD) and accuracy (%RE) ranged from 3.65 to 10.64 and from -2.14 to 3.07, respectively. Modifications to an existing assay for the determination of OLZ in human plasma were necessary. A different structural analog was used as the internal standard due to instability observed for the original analog when using human blood as the matrix. A second modification was the addition of the anti-oxidant sodium ascorbate to inhibit degradation of OLZ in human blood, as has been noted by other investigators. Upon fortification of human blood with sodium ascorbate (final concentration, 0.33 mM), OLZ was found to be stable for at least 1 week at -70 degrees C as well as through two freeze-thaw cycles. This assay, which will be used to investigate the distribution of OLZ in human blood, grants insight into the proper sample handling conditions needed to perform valid determinations of OLZ in human blood.

[Valgus tibial head reconstruction, monocondylar sled prosthesis or bicondylar gliding surface replacement in therapy of medial gonarthrosis--a cost analysis]. / Valgisierende Tibiakopfumstellung, monokondyläre Schlittenprothese oder bikondylärer Gleitflächenersatz zur Therapie der medialen Gonarthrose--eine Kostenstrukturanalyse.

AIM: Three to a certain extent competing methods for the treatment of unicompartmental osteoarthritis of the knee are compared regarding the costs they cause. These methods comprise the high tibial valgus osteotomy (HTO), the unicompartmental knee arthroplasty (UKA), and the total knee arthroplasty (TKA). METHODS: We compared the in-patient costs and out-patient costs of 20 patients (drawn by lot) in each group who received one of the above-named operative methods at our hospital between 1988 and 1993. The results were extrapolated according to the expected survival rate of the applied method. RESULTS: The average total costs of patients who received a HTO were 9.487,-; for the unicompartmental arthroplasty the average cost were 11.687,-; the implantation of a TKA resulted in average costs of 16.940,-. All operative procedures exceeded a particular global amount, the socalled "Fallpauschale". CONCLUSION: Regarding the total costs that arise for the operative treatment of the degenerative arthritis of the knee, the HTO (with or without hardware removal) proves to be the cheapest of the applied methods. Projecting the results on the survival rate of each treatment method, no statistically significant differences could be recognized between the three operative procedures.