Elevated baseline serum PD-1 or PD-L1 predicts poor outcome of PD-1 inhibition therapy in metastatic melanoma.

BACKGROUND: Programmed cell death protein 1 (PD-1) checkpoint inhibition has recently advanced to one of the most effective treatment strategies in melanoma. Nevertheless, a considerable proportion of patients show upfront therapy resistance and baseline predictive biomarkers of treatment outcome are scarce. In this study we quantified PD-1 and programmed death-ligand 1 (PD-L1) in baseline sera from melanoma patients in relation to therapy response and survival. PATIENTS AND METHODS: Sera taken at therapy baseline from a total of 222 metastatic melanoma patients (two retrospectively selected monocentric discovery cohorts, n = 130; one prospectively collected multicentric validation cohort, n = 92) and from 38 healthy controls were analyzed for PD-1 and PD-L1 concentration by sandwich enzyme-linked immunosorbent assay. RESULTS: Melanoma patients showed higher serum concentrations of PD-1 (P = 0.0054) and PD-L1 (P < 0.0001) than healthy controls. Elevated serum PD-1 and PD-L1 levels at treatment baseline were associated with an impaired best overall response (BOR) to anti-PD-1 (P = 0.014, P = 0.041), but not to BRAF inhibition therapy. Baseline PD-1 and PD-L1 serum levels correlated with progression-free (PFS; P = 0.0081, P = 0.053) and overall survival (OS; P = 0.055, P = 0.0062) in patients who received anti-PD-1 therapy, but not in patients treated with BRAF inhibitors. By combining both markers, we obtained a strong discrimination between favorable and poor outcome of anti-PD-1 therapy, with elevated baseline serum levels of PD-1 and/or PD-L1 associated with an impaired BOR (P = 0.037), PFS (P = 0.048), and OS (P = 0.0098). This PD-1/PD-L1 combination serum biomarker was confirmed in an independent multicenter validation set of serum samples prospectively collected at baseline of PD-1 inhibition (BOR, P = 0.019; PFS, P = 0.038; OS, P = 0.022). Multivariable Cox regression demonstrated serum PD-1/PD-L1 as an independent predictor of PFS (P = 0.010) and OS (P = 0.003) in patients treated with PD-1 inhibitors. CONCLUSION: Our findings indicate PD-1 and PD-L1 as useful serum biomarkers to predict the outcome of PD-1 inhibition therapy in melanoma patients and to select patients for PD-1-based versus BRAF-based therapy strategies.

[Study protocol of the monocentric prospective randomized placebo-controlled double-blind phase II study to explore the protective effects of the ginkgo biloba extract EGb 761® from temporary noise-induced hearing loss]. / Prüfplan für die monozentrische prospektive randomisierte placebokontrollierte doppelblinde Phase-II-Studie zur Exploration protektiver Effekte des Ginkgo-biloba-Spezialextrakts EGb 761® vor einer vorübergehenden Hörschädigung durch Lärm.

BACKGROUND: Hearing loss is frequently induced by occupational noise exposure and leads to rising hearing thresholds as well as reduced otoacoustic emissions (OAE), mostly caused by metabolic hair cell decompensation. OBJECTIVE: Primary endpoint is the increase in average pure tone thresholds after noise exposure, secondary endpoints are loss of distortion product and click-evoked OAE as well as reduction of their contralateral suppression. PARTICIPANTS AND METHODS: The present study design describes the verification of the anti-oxidant and neuroprotective properties of EGb 761® by evaluation of cochlear protection from noise impact as well as its safety and tolerance in 202 healthy male participants distributed equally to verum and placebo groups in a double-blind manner. Participants were assessed, medicated, exposed to noise, and then examined at timepoints up to 10 min and 4 weeks thereafter. CONCLUSION: This summary of the verification study protocol highlights the complexity of diligent and precise planning according to the European Medicines Agency criteria for controlled trials (EudraCT). Key points are the intervention rationale, definitions of in- and exclusion criteria, estimation of subject numbers, and examination method setting in terms of optimum endpoint description.

Ultrasonographic findings in two sheep with enzootic calcinosis.

INTRODUCTION: This report describes 2 sheep with enzootic calcinosis characterized by abnormal cardiovascular and respiratory findings and ascites causing abdominal distension. Both sheep were anorexic and listless and had increased heart and respiratory rates. Auscultation of the heart revealed a gallop rhythm in sheep 1 and a loud systolic heart murmur in sheep 2. The activities of liver enzymes were severely increased in both sheep. Abdominal ultrasonography showed severe ascites and congestion of the liver and caudal vena cava. Echocardiography in sheep 2 showed hyperechoic and markedly thickened mitral and aortic valves with moderate-severe mitral insufficiency and generalized cardiomegaly. Both sheep were euthanized and examined postmortem. In addition to ascites and pleural effusion, the principal lesions were nodular thickening of the heart valves and calcification of the aorta and other arteries. Nutrition of the sheep did not include hay pellets, but the sheep were kept together with alpacas and lamas and had access to the hay pellets of these animals. In addition visitors were allowed to feed the sheep with hay pellets offered by the zoo in a dispenser. The two types of hay pellets had Vit D concentrations of 9'900 IU VitD3/kg and 7'000 IU Vit D2/kg, respectively. The definitive diagnosis was enzootic calcinosis.

[Reha Aftercare--One Size Fits All? Analyses of Rehab Patients which do not Benefit from Reha Aftercare]. / Reha-Nachsorge - one size fits all? Analyse der Rehabilitanden, die nicht vom Reha-Nachsorgekonzept "neues Credo" profitieren.

BACKGROUND: In an own research project fund-ed by this program rehabilitation aftercare as part of a revised rehabilitation philosophy ("new credo") was tested and evaluated in a controlled longitudinal trial. Rehabilitation patients from the intervention group reported significantly better implementation of rehabilitation contents and objectives in everyday life during 12 months after their rehabilitation stay. Better long-term effects were also detectable. Anyway, not all rehabilitation patients seemed to benefit from the intervention. The present analysis focuses on this subgroup: How do rehabilitation patients that do not benefit from the "new credo" differ from rehabilitation patients that draw advantage from it? METHOD: Data from a controlled study were used for secondary analysis. 3 clinics implemented the "new credo". Primary outcomes were: participation constraints (IMET) and functional disabilities in everyday life (FFbH-R). The intervention clinics recruited a total of 166 rehabilitation patients. Data from 163 cases could be analyzed using quantitative methods. In addition, interview data from 7 unsuccessful rehabilitation patients was analyzed qualitatively. RESULTS: 102 (63%) rehabilitation patients improved their values in FFbH-R or IMET by at least 0.3 effect sizes (successful group). Among the 61 unsuccessful rehabilitation patients, 17 (10%) started rehabilitation with initial values that high that they could not reach the fixed minimal improvement criteria ("low burdened" group). They were excluded from further analysis. The remain-ing 44 (27%) rehabilitation patients constitute the "unsuccessful" group.Both groups have comparable levels of impairment concerning somatic and psychosocial parameters.At the end of the rehabilitation stay the "unsuccessful" group reached improvements of moderate to large effect sizes concerning the recorded health parameters. One year after rehabilitation "unsuccessful" rehabilitation patients fall back to their baseline levels of impairment, while the successful maintain their achieved effects after rehabilita-tion. The 2 groups rated several aspects of the intervention differently and the "unsuccessful" group rated the preparation for the time after the rehabilitation worse.The number of perceived aftercare elements proves to be the strongest predictor of failure in multivariate evaluation; "clinic facilities" is the only other independent predictor.During the interviews, "unsuccessful" patients reported different individual barriers for implementation of physical activity in daily life. CONCLUSIONS: Rehabilitation patients, who did not benefit from the new rehabilitation and aftercare concept, did neither differ in sociodemographic nor in disease-specific characteristics from the successful rehabilitation patients. However, differences in the implementation of the new rehabilitation philosophy and its individual components in the rehabilitation clinics are significantly associated with the occurrence of success or failure. Ov-er-all, the results of our study indicate a need for more flexible design of rehabilitation and aftercare adjusted to individual needs.

[Supported own initiative of rehabilitation patients ("new credo") - longterm effects of the nonrandomized trial]. / Begleitete Eigeninitiative nach der Reha ("Neues Credo") - Langzeitergebnisse der quasiexperimentellen Interventionsstudie.

OBJECTIVE: Our project is closely related to the previous study "Rehabilitation aftercare - a new credo for rehabilitation clinics"; the intensified aftercare-strategy was developed by an interdisciplinary group of health professionals and scientists for patients with chronic back pain. Main objective is the resumption and maintenance of regular physical activity in everyday life of pa-tients undergoing rehabilitation. The aim of our present study was to evaluate the long-term (24-month-follow-up) efficacy of the "New Credo" (inpatient medical rehabilitation+intensified aftercare=IG) as compared with usual care (inpatient medical rehabilitation=CG). METHODOLOGY: We conducted a prospective controlled multicentre trial (IG: 3 clinics vs. CG: 3 clinics) with 4 points of measurement (T0: before and T1: after rehabilitation as well as T3: 12 and T4: 24 month after discharge). We enrolled rehabilitants with a primary diagnosis of chronic back pain (ICD-10: M51-54) who had given their consent. Exclusion criteria were mental and/or psychological impairments as well as surgery in the past 6 months. Primary Outcomes included restriction of social participation (IMET) and functional capacity (FFbH-R). Treatment effects were evaluated using analyses of variance and adjusting for differences between baseline -values; furthermore we calculated effect sizes (SMDInter, SMDIntra, ORs). RESULTS: Complete data were available for 123 IG-rehabilitants and 299 CG-participants (Completer-Analysis, response: 52%). Regarding restriction in functional capacity in everyday life (FFbH-R) multivariate analysis confirmed a treatment*time interaction, the appropriate effect-sizes showed no significant effects (SMDInter and SMDIntra). With respect to long-term effects 24 months after discharge no significant effects were found for social participation, however. With respect to secondary outcomes there are no significant differences between the groups (SMDInter, ORs); in fact rehabilitants of both groups showed similar effects (SMDIntra) in most cases. CONCLUSION: In summary our results doesn't confirm that the intensified aftercare-programme is able to improve longer-term effectiveness of inpatient orthopaedic rehabilitation up to 24 month.

[Innovative 3D imaging]. / Innovative 3D-Bildgebung.

Intraoperative 2D fluoroscopy is often performed for repositioning and implant control. However, this does not always provide the details needed to reliably detect joint steps or incorrect repositioning. Over the last few years, intraoperative 3D imaging has been established and further developed. Multiple studies demonstrate an advantage and better intraoperative control through 3D imaging. Examples are the upper ankle, the proximal tibia and the distal radius; the rates of intraoperative revisions with digital volume tomography (DVT) are between 20-30%. Technical advancements, such as metal artifact reductions, automated plane setting, automated screw detection, and robotic DVT devices, facilitate intraoperative operation, shorten surgical time, and provide improved image quality. By processing the data sets in the form of an immersive, computer-simulated image in terms of "augmented reality" (AR), increased precision can be achieved intraoperatively while reducing radiation exposure. The implementation of these systems is associated with costs, which are offset by cost savings from avoided revisions. Adequate counter-financing is still lacking at the present time. Intraoperative 3D imaging represents an important tool for intraoperative control. The current data situation makes it necessary to address the routine use of 3D procedures, especially in the joint area. The indications are becoming increasingly broader. Technical innovations such as robotics and AR have significantly improved 3D devices in recent years and offer high potential for integration into the OR.

Concealed by darkness: Combination of NMR and HRMS reveal the molecular nature of dissolved organic matter in fractured-rock groundwater and connected surface waters.

Detailed molecular composition of solid phase extracted dissolved organic matter (SPEDOM) collected from fractured-rock groundwater was compared to connected surface river water at two different watersheds in the unconfined chalk aquifer of Champagne in France using full scan ultrahigh resolution electrospray and photoionization Fourier transform ion cyclotron mass spectrometry (FT-ICR MS), Orbitrap tandem MS (MS/MS) and 1H magnetic resonance spectroscopy (NMR). 1H NMR spectroscopy indicated that groundwater SPEDOM carried a higher contribution of aliphatic compounds while surface river waters SPEDOM were enriched in carboxyl-rich alicyclic molecules (CRAM), acetate derivatives and oxygenated units. Furthermore, we show here that use of photoionization (APPI(+)) in aquifer studies is key, ionizing about eight times more compounds than ESI in surface river water samples, specifically targeting the dissolved organic nitrogen pool, accounting for more than 50% of the total molecular space, as well as a non-polar, more aromatic fraction; with little overlap with compounds detected by ESI(-) FT-ICR MS. On the other hand, groundwater SPEDOM samples did not show similar selectivity as less molecular diversity was observed in APPI compared to ESI. Mass-difference transformation networks (MDiNs) applied to ESI(-) and APPI(+) FT-ICR MS datasets provided an overview of the biogeochemical relationships within the aquifer, revealing chemical diversity and microbial/abiotic reactions. Finally, the combination of ESI(-) FT-ICR MS and detailed Orbitrap MS/MS analysis revealed a pool of polar, anthropogenic sulfur-containing surfactants in the groundwaters, likely originating from agricultural runoff. Overall, our study shows that in this aquifer, groundwater SPEDOM contains a significantly reduced pool of organic compounds compared to surface river waters, possibly related to a combination of lack of sunlight and adsorption of high O/C formulas to mineral surfaces.

Impact of Anxiety During Hospitalization on the Clinical Outcome of Patients With Osteoporotic Thoracolumbar Vertebral Fracture.

STUDY DESIGN: Multicenter prospective cohort study. OBJECTIVES: Anxiety in combination with osteoporotic vertebral compression fractures (OVCFs) of the spine remains understudied. The purpose of this study was to analyze whether anxiety has an impact on the short-term functional outcome of patients with an OVCF. Furthermore, a direct impact of the fracture on the patient's anxiety during hospitalization should be recognized. METHODS: All inpatients with an OVCF of the thoracolumbar spine from 2017 to 2020 were included. Trauma mechanism, analgetic medication, anti-osteoporotic therapy, timed-up-and-go test (TuG), mobility, Barthel index, Oswestry-Disability Index (ODI) and EQ5D-5L were documented.For statistical analysis, the U test, chi-square independence test, Spearman correlation, General Linear Model for repeated measures, Bonferroni analysis and Wilcoxon test were used. The item anxiety/depression of the EQ5D-5L was analyzed to describe the patients' anxiousness. RESULTS: Data from 518 patients from 17 different hospitals were evaluated. Fracture severity showed a significant correlation (r = .087, P = .0496) with anxiety. During the hospital stay, pain medication (P < .001), anti-osteoporotic medication (P < .001), and initiation of surgical therapy (P < .001) were associated with less anxiety. The anxiety of a patient at discharge was negatively related to the functional outcomes at the individual follow-up: TuG (P < .001), Barthel index (P < .001), ODI (P < .001) and EQ5D-5L (P < .001). CONCLUSIONS: Higher anxiety is associated with lower functional outcome after OVCF. The item anxiety/depression of the EQ5D-5L provides an easily accessible, quick and simple tool that can be used to screen for poor outcomes and may also offer the opportunity for a specific anxiety intervention.

Intrinsic instability of aberration-corrected electron microscopes.

Aberration-corrected microscopes with subatomic resolution will impact broad areas of science and technology. However, the experimentally observed lifetime of the corrected state is just a few minutes. Here we show that the corrected state is intrinsically unstable; the higher its quality, the more unstable it is. Analyzing the contrast transfer function near optimum correction, we define an "instability budget" which allows a rational trade-off between resolution and stability. Unless control systems are developed to overcome these challenges, intrinsic instability poses a fundamental limit to the resolution practically achievable in the electron microscope.

[Supported own initiative of rehabilitation patients ("new credo") - a successful model?]. / Begleitete Eigeninitiative nach der Reha ("neues Credo") - ein Erfolgsmodell?

BACKGROUND: In recent years, rehabilitation aftercare has become an important topic in rehabilitation research and practice. This development was initiated by the lack of long-term effects after rehabilitation treatment and by the deficits identified in several reviews of aftercare recommendations and of actual practice of aftercare. Against this background, a new aftercare strategy was developed, the "New Credo", which focused aftercare from the beginning of rehabilitation treatment. A substantial element of the concept is the resumption and maintenance of physical activity in everyday life of the rehabilitation patient. The goal of the present study was the evaluation of this concept in a sample of patients with chronic back pain. METHOD: The "New Credo" was evaluated in a multi-centre, prospective controlled longitudinal study with 3 points of measurement (before and after rehab as well as after 12 months). 6 rehabilitation clinics in Schleswig-Holstein were included in the intervention arm (IG, 3 clinics) and in the control arm (CG, 3 clinics), participants were rehabilitants with chronic back pain. The clinics in the intervention arm implemented "the New Credo" in accordance with their local conditions. During the study period they were provided with a person who was responsible for managing aftercare activities for patients. Participants in the CG received standard rehabilitation treatment according to general guidelines. Restriction of social participation, disability days and restriction in functional capacity in everyday life were defined as primary outcomes. Complete data were available for 166 rehabilitants in the IG and 368 in the CG. RESULTS: At the end of rehabilitation rehabilitants in the IG reported significantly higher use of treatment offerings and more intensive preparation for the time after rehabilitation treatment than rehabilitants in the CG. Rehabilitants in the IG found recommendations for the time after treatment significantly better than those in the CG. As expected, both rehabilitants in the IG and in the CG showed similar outcomes directly after inpatient treatment. With respect to long-term effects 12 months later, significant differences with moderate effect sizes were observed in 2 of the 3 primary outcomes (function capacity and social participation) in favour of the IG. With respect to secondary outcomes, rehabilitants in the IG showed significantly better long-term results than those in the CG. DISCUSSION AND CONCLUSION: Our results show that the "New Credo" can successfully be implemented into rehabilitation practice. The "New Credo" was positively rated by clinicians and rehabilitants. Rehabilitants in the IG found offerings of supported aftercare very helpful. The intensity of actual use of aftercare by rehabilitants as well as the emphasis laid on aftercare by clinicians indicate that treatment was focused on aftercare over the whole time of rehabilitation and that the rehabilitants transferred these aspects into everyday life. For the rehabilitants in the IG higher long-term effects could be observed compared to those in the CG. This suggests an improved sustainability of rehabilitation effects and a successful transfer of acquired knowledge and self-efficacy in everyday life.

Outcome and quality of life favour a conservative treatment of patients with primary gastric lymphoma.

BACKGROUND: Both surgical and conservative treatment strategies offer an excellent outcome to patients with gastric lymphoma. No comparison of surgical resection with primary radiation or chemotherapy on a prospective randomised basis was available up to now. AIMS: The aim of this study was to compare surgery with radiation and surgery plus chemotherapy with chemotherapy alone in patients with marginal zone B-cell lymphoma (MZBCL) of MALT and diffuse large B-cell lymphoma (DLBCL), respectively, with respect to long-term outcome and quality of life. METHODS: 49 patients [21 male and 28 female; age 65 (35 - 75) years] with newly diagnosed MZBCL of MALT (n = 19) and DLBCL (n = 30) of stage I and II were recruited. Outcome was evaluated after a follow-up of 74 (7 - 102) months and quality of life was measured using the SF-36 health survey and the gastrointestinal life quality index (GLQI). RESULTS: Complete remission rates in MZBCL of MALT were 88 % and 89 % in patients treated by surgery or radiotherapy, respectively. The corresponding rates for patients with DLBCL were 93 % following surgery plus chemotherapy and 92 % after chemotherapy alone, respectively. The results in the different treatment groups did not differ significantly. Posttherapeutic quality of life was generally high with a significant advantage for patients treated conservatively compared to those who underwent surgery. CONCLUSION: Long-term outcome of patients with gastric lymphoma is excellent irrespective of the lymphoma type and the treatment approach. Considering quality of life a conservative therapeutic strategy should be favoured.

Synaptonemal complex protein SYCP3 exists in two isoforms showing different conservation in mammalian evolution.

Meiosis-specific protein SYCP3 is a major structural component of synaptonemal complex (SC) lateral elements. SYCP3 is rather well conserved in vertebrates. However, some differences in SYCP3 expression have been shown among mammals. To clarifiy these differences, we have investigated SYCP3 expression with the aid of bioinformatic, biochemical and cell biological methods. Here we show that, in contrast to other vertebrates, rat and mouse SYCP3 exist in 2 isoforms. The short isoform is conserved among vertebrates. However, the longer isoform, which represents an N-terminal extension of the shorter one, most likely appeared about 15 million years ago in a common ancestor of rat and mouse and after separation from the hamster branch.

High-avidity human IgG kappa monoclonal antibodies from a novel strain of minilocus transgenic mice.

Human immunoglobulin transgenic mice provide a method of obtaining human monoclonal antibodies (Mabs) using conventional hybridoma technology. We describe a novel strain of human immunoglobulin transgenic mice and the use of this strain to generate multiple high-avidity human sequence IgG kappa Mabs directed against a human antigen. The light chain transgene is derived in part from a yeast artificial chromosome clone that includes nearly half of the germline human V kappa region. In addition, the heavy-chain transgene encodes both human mu and human gamma 1 constant regions, the latter of which is expressed via intratransgene class switching. We have used these animals to isolate human IgG kappa Mabs that are specific for the human T-cell marker CD4, have high binding avidities, and are immunosuppressive in vitro. The human Mab-secreting hybridomas display properties similar to those of wild-type mice including stability, growth, and secretion levels. Mabs with four distinct specificities were derived from a single transgenic mouse, consistent with an extensive diversity in the primary repertoire encoded by the transgenes.

[Pneumothorax during right-sided nephrectomy in a heifer]. / Pneumothorax als Komplikation einer rechtsseitigen Nephrektomie bei einem Jungrind.

A 250-kg heifer had signs of colic attributable to urolithiasis of the right kidney. Medical treatment did not result in resolution of clinical signs, and nephrectomy was carried out. The surgery was started with the heifer standing, and the 13th rib was resected. However, during blunt dissection of the kidney, air suddenly entered the pleural space and the heifer had acute severe dyspnoea. The hole in the pleural cavity was sutured and a chest drain was placed. Inhalation anaesthesia was then induced and nephrectomy could be completed without further complications. The heifer was discharged 11 days postoperatively, and was healthy and had been integrated into the herd 12 months after surgery. Pneumothorax must be considered a possible complication of rib resection in right-sided nephrectomy in cattle.

Reconciling simulated melting and ground-state properties of metals with a modified embedded-atom method potential.

We propose a modification of the embedded-atom method-type potential aiming at reconciling simulated melting and ground-state properties of metals by means of classical molecular dynamics. Considering titanium, magnesium, gold, and platinum as case studies, we demonstrate that simulations performed with the modified force field yield quantitatively correctly both the melting temperature of the metals and their ground-state properties. It is shown that the accounting for the long-range interatomic interactions noticeably affects the melting point assessment. The introduced modification weakens the interaction at interatomic distances exceeding the equilibrium one by a characteristic vibration amplitude defined by the Lindemann criterion, thus allowing for the correct simulation of melting, while keeping its behavior in the vicinity of the ground state minimum. The modification of the many-body potential has a general nature and can be applicable to metals with different characteristics of the electron structure as well as for many different molecular and solid state systems experiencing phase transitions.

Quantitative electron phase imaging with high sensitivity and an unlimited field of view.

As it passes through a sample, an electron beam scatters, producing an exit wavefront rich in information. A range of material properties, from electric and magnetic field strengths to specimen thickness, strain maps and mean inner potentials, can be extrapolated from its phase and mapped at the nanoscale. Unfortunately, the phase signal is not straightforward to obtain. It is most commonly measured using off-axis electron holography, but this is experimentally challenging, places constraints on the sample and has a limited field of view. Here we report an alternative method that avoids these limitations and is easily implemented on an unmodified transmission electron microscope (TEM) operating in the familiar selected area diffraction mode. We use ptychography, an imaging technique popular amongst the X-ray microscopy community; recent advances in reconstruction algorithms now reveal its potential as a tool for highly sensitive, quantitative electron phase imaging.

Rational design of an animal model for Alzheimer's disease: introduction of multiple human genomic transgenes to reproduce AD pathology in a rodent.

A major obstacle to understanding the pathogenesis of Alzheimer's disease is the lack of easily studied animal models. Our approach is to apply transgenic methods to humanize mice and rats, employing methods that introduce large genomic transgenes, because this improves the level of transgene protein expression and the tissue specificity of expression. Our plan is to reproduce AD pathology in rodents by making them transgenic for several human proteins involved in AD. This report describes transgenic animal lines that we have produced, and summarizes our current approach and future plans. Two human genes known to be involved in AD pathology are the amyloid precursor protein (APP) and the E4 isoform of apolipoprotein E (apoE4). So far, we have produced and analyzed a transgenic line carrying the entire human APP gene cloned in a yeast artificial chromosome. We have also produced but not yet analyzed a mouse carrying the human apoE4 gene. Work is in progress to produce a transgenic line carrying a disease-causing mutation in the human APP gene. As we produce these animals, we are breeding them together, and also breeding them with a mouse line that lacks endogenous apoE, to produce an animal model carrying several human proteins whose interaction is believed to be instrumental in development of AD pathology. These transgenic animals will be useful for dissecting the biochemical and physiological steps leading to AD, and for development of therapies for disease intervention.

Methylxanthines inhibit glucose transport in rat adipocytes by two independent mechanisms.

In summary, this study characterized the biphasic inhibition of fat cell glucose transport by the lipolytic agents caffeine and theophylline. Like the lipolytic drug forskolin, both methylxanthines produced an immediate inhibition of glucose transport that was not seen with 8-phenyltheophylline, a pure adenosine receptor antagonist. The immediate inhibition was therefore not mediated by the adenosine receptor antagonism but seems to be due to a direct interaction with the hexose transporter. This conclusion is supported by the immediate onset of the inhibition and additionally by the interference of theophylline and caffeine with the binding of cytochalasin B, a ligand of the glucose transporter that binds to an intracellular site of the transporter molecule. In addition, a second, delayed inhibitory effect of theophylline and caffeine on glucose transport was observed. This portion shared many aspects of the inhibitory effect of lipolytic hormones. It developed over a period of about 5 min and was antagonized by the simultaneous addition of the antilipolytic hormone PGE2. This component of transport inhibition could be attributed to the antagonistic effect of methylxanthines at the fat cell A1-adenosine receptor since it was also seen with 8-phenyltheophylline. This conclusion is further supported by data showing that the removal of endogenous adenosine with adenosine deaminase resulted in a comparable 25-30% inhibition of insulin-stimulated glucose transport. In addition, the time course of glucose transport inhibition by the subsequent addition of adenosine deaminase is similar to that of the delayed portion of the inhibition seen with theophylline and caffeine. Both treatments produced their maximal inhibition after 5 min. In conclusion, the methylxanthines theophylline and caffeine inhibit glucose transport by a combination of two different modes of action. The immediate major component is mediated via a direct interaction with the hexose transporter whereas the delayed component involves adenosine receptor antagonism and thereby the interaction with G-proteins.

Prostaglandin E2 differentiates between two forms of glucose transport inhibition by lipolytic agents.

The inhibition of insulin-stimulated glucose transport by lipolytic agents was studied in isolated rat adipose cells. Two different mechanisms for the inhibition of glucose transport by lipolytic hormones and agents were distinguished by use of the antilipolytic agent prostaglandin E2 (PGE2). The inhibition of glucose transport induced by lipolytic hormones such as glucagon, catecholamines or ACTH in the presence of adenosine deaminase was antagonized by PGE2. In contrast, inhibition of hexose transport by alkylxanthines was only partially (20-30%) attenuated by PGE2, although the eicosanoid had antagonized cyclic AMP accumulation and stimulation of lipolysis in response to all tested lipolytic agents. The inhibition of glucose transport by IBMX was immediate, whereas the lipolytic hormones (isoprenaline and ACTH) exhibited a latency of 2-3 min. In addition, the inhibition induced by the lypolytic hormones disappeared after cooling of the cells to 22 degrees C, at which temperature IBMX was still inhibitory. Thus, the PGE2-sensitive component of the effect of lipolytic agents on glucose transport appears to be mediated by adenylate cyclase or its subunits Ns/Ni. The PGE2-insensitive effect of alkylxanthines probably reflects a direct interaction of the agents with a regulatory site at the transporter or a related protein.

Developing tools for better medical management of Medicare populations.

Medicare enrollment in HMOs got off to a quick start but has ebbed recently, principally because health plans found that the elderly joining their plans were costing the plan more than what the federal government was willing to pay. For the MCOs that want to continue to participate in Medicare + Choice, it is imperative for them to understand, in as much detail as possible, the potential health care costs of their enrollees.