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Human aging is marked by the emergence of a tapestry of clonal expansions in dividing tissues, particularly evident in blood as clonal hematopoiesis (CH). CH, linked to cancer risk and aging-related phenotypes, often stems from somatic mutations in a set of established genes. However, the majority of clones lack known drivers. Here we infer gene-level positive selection in whole blood exomes from 200,618 individuals in UK Biobank. We identify 17 additional genes, ZBTB33, ZNF318, ZNF234, SPRED2, SH2B3, SRCAP, SIK3, SRSF1, CHEK2, CCDC115, CCL22, BAX, YLPM1, MYD88, MTA2, MAGEC3 and IGLL5, under positive selection at a population level, and validate this selection pattern in 10,837 whole genomes from single-cell-derived hematopoietic colonies. Clones with mutations in these genes grow in frequency and size with age, comparable to classical CH drivers. They correlate with heightened risk of infection, death and hematological malignancy, highlighting the significance of these additional genes in the aging process., (© 2024. The Author(s).)

Immunological targeting of pathological cells has been successful in oncology and is expanding to other pathobiological contexts. Here, we present a flexible platform that allows labeling cells of interest with the surface-expressed model antigen ovalbumin (OVA), which can be eliminated via either antigen-specific T cells or newly developed OVA antibodies. We demonstrate that hepatocytes can be effectively targeted by either modality. In contrast, pro-fibrotic fibroblasts associated with pulmonary fibrosis are only eliminated by T cells in initial experiments, which reduced collagen deposition in a fibrosis model. This new experimental platform will facilitate development of immune-based approaches to clear potential pathological cell types in vivo., (© 2023 Zhang et al.)

The spleen plays a key role in iron homeostasis. It is the largest filter of the blood and performs iron reuptake from old or damaged erythrocytes. Despite this role, spleen iron concentration has not been measured in a large, population-based cohort. In this study, we quantify spleen iron in 41,764 participants of the UK Biobank by using magnetic resonance imaging and provide a reference range for spleen iron in an unselected population. Through genome-wide association study, we identify associations between spleen iron and regulatory variation at two hereditary spherocytosis genes, ANK1 and SPTA1. Spherocytosis-causing coding mutations in these genes are associated with lower reticulocyte volume and increased reticulocyte percentage, while these common alleles are associated with increased expression of ANK1 and SPTA1 in blood and with larger reticulocyte volume and reduced reticulocyte percentage. As genetic modifiers, these common alleles may explain mild spherocytosis phenotypes that have been observed clinically. Our genetic study also identifies a signal that co-localizes with a splicing quantitative trait locus for MS4A7, and we show this gene is abundantly expressed in the spleen and in macrophages. The combination of deep learning and efficient image processing enables non-invasive measurement of spleen iron and, in turn, characterization of genetic factors related to the lytic phase of the erythrocyte life cycle and iron reuptake in the spleen., Competing Interests: Declaration of interests E.P.S., R.L.C., and M.C. are employees of Calico Life Sciences LLC. Y.L. is a former employee of Calico Life Sciences LLC., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Nicotinamide adenine dinucleotide (NAD) levels decline during aging, contributing to physical and metabolic dysfunction. The NADase CD38 plays a key role in age-related NAD decline. Whether the inhibition of CD38 increases lifespan is not known. Here, we show that the CD38 inhibitor 78c increases lifespan and healthspan of naturally aged mice. In addition to a 10% increase in median survival, 78c improved exercise performance, endurance, and metabolic function in mice. The effects of 78c were different between sexes. Our study is the first to investigate the effect of CD38 inhibition in naturally aged animals., (© 2022 The Authors. Aging Cell published by Anatomical Society and John Wiley & Sons Ltd.)

The process wherein dividing cells exhaust proliferative capacity and enter into replicative senescence has become a prominent model for cellular aging in vitro. Despite decades of study, this cellular state is not fully understood in culture and even much less so during aging. Here, we revisit Leonard Hayflick's original observation of replicative senescence in WI-38 human lung fibroblasts equipped with a battery of modern techniques including RNA-seq, single-cell RNA-seq, proteomics, metabolomics, and ATAC-seq. We find evidence that the transition to a senescent state manifests early, increases gradually, and corresponds to a concomitant global increase in DNA accessibility in nucleolar and lamin associated domains. Furthermore, we demonstrate that senescent WI-38 cells acquire a striking resemblance to myofibroblasts in a process similar to the epithelial to mesenchymal transition (EMT) that is regulated by t YAP1/TEAD1 and TGF-β2. Lastly, we show that verteporfin inhibition of YAP1/TEAD1 activity in aged WI-38 cells robustly attenuates this gene expression program., Competing Interests: MC, HY, IS, TM, RW, AI, JO, JG, LC, TV, AF, CK, BB, FM, DK, MR, RC, AL, DB, DH is affiliated with Calico Life Sciences, LLC. The author has no other competing interests to declare, (© 2022, Chan et al.)

Macaques with self-injurious behavior (SIB) have been used as a model of human SIB and have previously been shown to respond to treatments targeting enhancement of central serotonin signaling, whether by supplementation with tryptophan, or by inhibiting synaptic reuptake. Decreased serotonin signaling in the brain has also been implicated in many human psychopathologies including major depression disorder. A disturbance in tryptophan metabolism that moves away from the production of serotonin and toward the production of kynurenine has been proposed as a major etiological factor of depression. We hypothesized that in macaques with SIB, central tryptophan metabolism would be shifted toward kynurenine production, leading to lower central serotonin (5-hydroxytryptamine). We analyzed tryptophan metabolites in the cerebral spinal fluid (CSF) of macaques with and without SIB to determine whether and where tryptophan metabolism is altered in affected animals as compared with behaviorally normal controls. We found that macaques with SIB had lower CSF concentrations of serotonin than did behaviorally normal macaques, and that these deficits were inversely correlated with the severity of abnormal behavior. However, our results suggest that this decrease is not due to shifting of the tryptophan metabolic pathway toward kynurenine, as concentrations of kynurenine were also low. Concentrations of IL6 were elevated, suggesting central inflammation. Determining the mechanism by which serotonin function is altered in self-injurious macaques could shed light on novel therapies for SIB and other disorders of serotonin signaling.

Introduction: Knowledge of the contemporary epidemiology of hepatitis B virus (HBV) infection among military personnel can inform potential Department of Defense (DoD) screening policy and infection and disease control strategies., Materials and Methods: HBV infection status at accession and following deployment was determined by evaluating reposed serum from 10,000 service members recently deployed to combat operations in Iraq and Afghanistan in the period from 2007 to 2010. A cost model was developed from the perspective of the Department of Defense for a program to integrate HBV infection screening of applicants for military service into the existing screening program of screening new accessions for vaccine-preventable infections., Results: The prevalence of chronic HBV infection at accession was 2.3/1,000 (95% CI: 1.4, 3.2); most cases (16/21, 76%) identified after deployment were present at accession. There were 110 military service-related HBV infections identified. Screening accessions who are identified as HBV susceptible with HBV surface antigen followed by HBV surface antigen neutralization for confirmation offered no cost advantage over not screening and resulted in a net annual increase in cost of $5.78 million. However, screening would exclude as many as 514 HBV cases each year from accession., Conclusions: Screening for HBV infection at service entry would potentially reduce chronic HBV infection in the force, decrease the threat of transfusion-transmitted HBV infection in the battlefield blood supply, and lead to earlier diagnosis and linkage to care; however, applicant screening is not cost saving. Service-related incident infections indicate a durable threat, the need for improved laboratory-based surveillance tools, and mandate review of immunization policy and practice., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Association of Military Surgeons of the United States.)

Introduction: Some health determinants require relatively stronger health system capacity and socioeconomic development than others to impact child mortality. Few quantitative analyses have analyzed how the impact of health determinants varies by mortality level., Methods: 149 low- and middle-income countries were stratified into high, moderate, low, and very low baseline levels of child mortality in 1990. Data for 52 health determinants were collected for these countries for 1980-2010. To quantify how changes in health determinants were associated with mortality decline, univariable and multivariable regression models were constructed. An advanced statistical technique that is new for child mortality analyses-MM-estimation with first differences and country clustering-controlled for outliers, fixed effects, and variation across decades., Findings: Some health determinants (immunizations, education) were consistently associated with child mortality reduction across all mortality levels. Others (staff availability, skilled birth attendance, fertility, water and sanitation) were associated with child mortality reduction mainly in low or very low mortality settings. The findings indicate that the impact of some health determinants on child mortality was only apparent with stronger health systems, public infrastructure and levels of socioeconomic development, whereas the impact of other determinants was apparent at all stages of development. Multisectoral progress was essential to mortality reduction at all baseline mortality levels., Conclusion: Policy-makers can use such analyses to direct investments in health and non-health sectors and to set five-year child mortality targets appropriate for their baseline mortality levels and local context.

Purpose of Review: Spinocerebellar ataxia type 12 (SCA12) is a rare autosomal dominant neurodegenerative disease characterized by tremor, gait abnormalities, and neuropsychiatric syndromes. The location of the causative CAG/CTG expansion mutation in PPP2R2B, a gene encoding regulatory units of the protein phosphatase 2A, may provide unique insights into the pathogenesis of neurodegeneration., Recent Findings: The first neuropathological examination of a brain from an SCA12 patient revealed both cerebellar and cerebral cortical atrophy, with a noted loss of Purkinje cells and no evidence of polyglutamine aggregates. Molecular investigations have demonstrated considerable complexity of PPP2R2B, which appears to encode at least eight isoforms each with a different N-terminal region. The repeat potentially influences PPP2R2B expression, and is itself included in several splice variants, falling within an open reading frame of at least one of these variants., Summary: The current data suggest at least two nonmutually exclusive hypotheses of SCA12 neurodegeneration. First, the repeat may influence PPP2R2B expression, by altering promoter activity, splicing, or transcript stability. This hypothesis would predict that the mutation changes the regulation of protein phosphatase 2A, with implications for the phosphoproteome. Alternatively, the repeat itself may be expressed and have toxic properties, though perhaps not through polyglutamine tracts. Either hypothesis may provide novel insight into the pathogenesis of neurodegeneration.

The discovery and development of new medicines that promote human health and potentially extend natural life remains a remarkably challenging endeavor. In this Commentary, we identify key elements of communication required to successfully translate promising biological findings to novel approved drug therapies and discuss the attendant challenges and opportunities., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Objective: SCA12 is a progressive autosomal-dominant disorder, caused by a CAG/CTG repeat expansion in PPP2R2B on chromosome 5q32, and characterized by tremor, gait ataxia, hyperreflexia, dysmetria, abnormal eye movements, anxiety, depression, and sometimes cognitive impairment. Neuroimaging has demonstrated cerebellar and cortical atrophy. We now present the neuropathology of the first autopsied SCA12 brain and utilize cell models to characterize potential mechanisms of SCA12 neurodegeneration., Methods: A fixed SCA12 brain was examined using gross, microscopic, and immunohistochemical methods. The effect of the repeat expansion on PPP2R2B Bβ1 expression was examined in multiple cell types by transient transfection of constructs containing the PPP2R2B Bβ1 promoter region attached to a luciferase reporter. The neurotoxic effect of PPP2R2B overexpression was examined in transfected rat primary neurons., Results: Neuropathological investigation revealed enlarged ventricles, marked cerebral cortical atrophy and Purkinje cell loss, less-prominent cerebellar and pontine atrophy, and neuronal intranuclear ubiquitin-positive inclusions, consistent with Marinesco bodies, which did not stain for long polyglutamine tracts, alpha-synuclein, tau, or transactive response DNA-binding protein 43. Reporter assays demonstrated that the region of PPP2R2B containing the repeat functions as a promoter, and that promoter activity increases with longer repeat length and is dependent on cell type, repeat sequence, and sequence flanking the repeat. Overexpression of PPP2R2B in primary cortical neurons disrupted normal morphology., Conclusions: SCA12 involves extensive, but selective, neurodegeneration distinct from Alzheimer's disease, synucleinopathies, tauopathies, and glutamine expansion diseases. SCA12 neuropathology may arise from the neurotoxic effect of repeat-expansion-induced overexpression of PPP2R2B., (© 2015 International Parkinson and Movement Disorder Society.)

Background: The health Millennium Development Goals (4, 5, 6) impose the same ambitious 2015 targets on every country. Few low-income countries are on track to reach them. Some authors have proposed country-specific targets as a more informative method by which countries can measure their progress against their potential., Methods: This paper demonstrates a supplementary approach to assess individual country progress that complements the global goals by adjusting for socioeconomic resources and prior time trends. A minimum performance target adjusts for time and national GDP. Fast-track targets, based on best-performing countries' progress within regional and income groups, adjust for health and non-health sector factors known to affect maternal and child health., Results: Measuring by the minimum performance target, 74% and 59% of low- and middle-income countries are on track for reducing child mortality and maternal mortality, respectively, compared with 69% and 22% using global MDGs. Only 20% and 7% of low- and middle-income countries are on track for the child and maternal mortality fast-track targets., Conclusions: Supplementary targets in maternal and child health, adjusted for each country's resources and policy performance can help countries know if they are truly underperforming relative to their potential. Adjusted targets can also flag countries that have surpassed their potential, and open opportunities for learning from success., Funding: Partnership for Maternal, Newborn & Child Health and the Alliance for Health Policy and Systems Research, as part of the Success Factors Study on reducing maternal and child mortality.

Rapidly evolving genome technology has enabled extensive molecular analysis of limited tumor biopsy material, thereby facilitating the broader implementation of personalized cancer medicine. However, genomics-based patient stratification across diverse tumor types is unlikely to supplant tissue-of-origin considerations in addressing clinical needs, including the development and application of novel "rationally targeted" cancer therapies., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: President's Emergency Plan for AIDS Relief (PEPFAR) has been criticized for its vertical or "stove-piping" structure, with resources targeting a specific disease rather than working to strengthen the underlying health system. This study aimed to evaluate whether PEPFAR activities were associated with system-wide improvements in both proximal and distal indicators of health systems strengthening., Methods: The World Bank database provided 12 indicators of health systems strengthening that were analyzed for their relationship to PEPFAR. Poisson and linear regression models were used to estimate the time trend. We evaluated the PEPFAR impact on health outcomes by comparing the time trend in each of the above indicators between 2 time periods: from 1995 to 2002 (pre-PEPFAR) and from 2004 to 2010 (during PEPFAR)., Results: PEPFAR activities showed strong and statistically significant associations with improvements in proximal indicators of health systems strengthening, those related directly to HIV, specifically life expectancy (P = 0.003) and tuberculosis incidence, prevalence, and mortality (P < 0.0001). There were nonsignificant associations between PEPFAR and improvements in distal indicators of health systems strengthening, namely infant mortality and under 5 child mortality., Conclusions: The progressive scale-up of PEPFAR-supported activities was associated with consistent improvements in proximal indicators of health systems strengthening. It was also associated with improvements in broader measures of health system strength, most clearly life expectancy. Given the limited number of health measures available for this type of analysis, more data must be collected for other indicators to evaluate the effectiveness of the many multibillion dollar global health initiatives.

Two of the great successes in the many decades-long 'war on cancer' are the emergence of adjuvant chemotherapy regimens with lifesaving potential and the subsequent wave of 'targeted' therapies addressing the unique vulnerabilities of particular tumor types. The first intersection of adjuvant treatment and targeted treatment resulted in a spectacularly positive outcome as the addition of the anti-HER2 humanized monoclonal antibody trastuzumab to the standard adjuvant chemotherapy essentially halved the relapse rate among women with HER2+ tumors. Subsequent studies of adjuvant trastuzumab have confirmed its dramatic efficacy in a variety of chemotherapeutic contexts and have been instructive in elucidating some of the challenges ahead for newer targeted agents. The recent negative experience with bevacizumab in the adjuvant colon cancer setting suggests pitfalls and limitations of the current approach to developing adjuvant regimens. A change in thinking may be required to gain the substantial benefits implied by the trastuzumab experience in the broader context of the targeted treatments. The case for a revitalized industry/academia/government partnership to address these challenges is compelling, with the potential for enormous patient and societal benefit. In order to bring potentially lifesaving benefits of this new generation of cancer drugs to patients more rapidly, changes to our 'war strategy' appear necessary.

The Flexner Report established guidelines for medical education and made the university the obligate home for medical education. Flexner mandated specific elements necessary for university-based premedical education. With the exception of the MCAT, much less attention has been paid to premedical education and its integration into the scope of medical education than to education within the confines of the medical school. This article reviews the history of premedical education, describes some recent critiques of premedical education, discusses a newer program for premedical education evolving at the University of Illinois at Chicago, and offers some suggestions for the future.

With 'efficiency savings' the watchword for health and social care services, reorganisation and labour rationalisation are the order of the day. This article examines the difficulties involved in (re)organising work which takes bodies as its object, or material of production. It shows that working on bodies ('body work') systematically delimits possibilities for labour process rationalisation which, in turn, constrains reorganisation of the health and social care sector. It does this in three main ways. First: rigidity in the ratio of workers to bodies-worked-upon limits the potential to increase capital-labour ratios or cut labour. Secondly: the requirement for co-presence and temporal unpredictability in demand for body work diminish the spatial and temporal malleability of the labour process. Thirdly: the nature of bodies as a material of production--complex, unitary and responsive--makes it difficult to standardise, reorganise or rationalise work. A wide-ranging analysis of body work in health and social care, as well as other sectors, fleshes out these three constraints and shows that attempts to overcome them and reorganise the sector in pursuit of cost savings or 'efficiency', generate problems for workers and the patients, whose bodies they work upon., (© 2011 The Author. Sociology of Health & Illness © 2011 Foundation for the Sociology of Health & Illness/Blackwell Publishing Ltd.)

Body work is a central activity in the practice of many workers in the field of health and social care. This article provides an introduction to the concept of body work--paid work on the bodies of others--and demonstrates its importance for understanding the activities of health and social care workers. Providing an overview of existing research on body work, it shows the manifold ways in which this can inform the sociology of health and illness--whether through a micro-social focus on the inter-corporeal aspects of work in health and social care, or through elucidating our understanding of the times and spaces of work, or through highlighting the relationship between mundane body work and the increasingly global movements of bodies, workers and those worked-upon. The article shows how understanding work undertaken on the bodies of others as 'body work' provides a mechanism for relating work in the sphere of health and social care to that in other sectors, opening up new avenues for research., (© 2011 The Authors. Sociology of Health & Illness © 2011 Foundation for the Sociology of Health & Illness/Blackwell Publishing Ltd.)

The catecholamine norepinephrine (NE) stimulates T lymphocytes through a beta-adrenergic receptor (βAR)/adenylyl cyclase (AC)/cyclic AMP (cAMP)/protein kinase A (PKA) pathway, leading to altered cell responsiveness and apoptosis. p38 Mitogen-activated protein kinase (MAPK), a major intracellular signalling mediator for cellular and environmental stressors, is involved in the production of immune modulators and in the regulation of T-cell development, survival and death. In these studies we investigated the relationship among NE signalling, p38 MAPK activity and T-cell death. We showed that NE stimulation of BALB/c mouse thymocytes and S49 thymoma cells selectively increases the dual phosphorylation and activity of p38α MAPK. p38 MAPK activation involves the βAR, Gs protein, AC, cAMP and PKA, as determined through the use of a βAR antagonist, activators of AC and cAMP, and S49 clonal mutants deficient in Gs and PKA. Dual phosphorylation of p38 MAPK is also dependent on its own catalytic activity. Inhibition of p38 MAPK activity revealed its involvement in cAMP-mediated activating transcription factor-2 (ATF-2) phosphorylation, Fas ligand messenger RNA (mRNA) up-regulation, and cell death. These results identify a mechanism through which NE stimulation of the βAR/Gs/PKA pathway activates p38 MAPK, which can be potentiated by autophosphorylation, and leads to changes in T-cell dynamics, in part through the regulation of Fas ligand mRNA expression.

The purpose of this study was to assess the pharmacokinetics of gabapentin in healthy greyhound dogs after single oral doses targeted at 10 and 20mg/kg PO. Six healthy greyhounds were enrolled (3 males, 3 females). Blood was obtained at predetermined times for the measurement of gabapentin plasma concentrations by liquid chromatography/mass spectrometry. Pharmacokinetic parameters were determined with computer software. The actual mean (and range) doses administered were 10.2 (9.1-12.0) mg/kg and 20.5 (18.2-24) mg/kg for the 10mg/kg and 20mg/kg targeted dose groups. The mean C(MAX) for the 10 and 20mg/kg groups were 8.54 and 13.22 microg/mL at 1.3 and 1.5h, and the terminal half-lives were 3.3 and 3.4h, respectively. The relative bioavailability of the 10mg/kg group was 1.13 compared to the 20mg/kg group. Gabapentin was rapidly absorbed and eliminated in dogs, indicating that frequent dosing is needed to maintain minimum targeted plasma concentrations., (2009 Elsevier Ltd. All rights reserved.)

The United States leads the world in creating prisoners, incarcerating one in 100 adults and housing 25% of the world's prisoners. Since the 1976, the US Supreme Court ruling that mandated health care for inmates, doctors have been an integral part of the correctional system. Yet conditions within corrections are not infrequently in direct conflict with optimal patient care, particularly for those suffering from mental illness and addiction. In addition to providing and working to improve clinical care for prisoners, physicians have an opportunity and an obligation to advocate for reform in the system of corrections when it conflicts with patient well-being.

This study evaluated the effects of a cysteinyl-leukotriene-1 (cys-LT(1)) receptor antagonist, zarfirlukast, during feline endotoxemia. Six adult, sexually intact male cats received either placebo or zarfirlukast (10mg, PO) and endotoxin (2 μg/kg/h q 6h) in a cross-over design. Rectal temperature, heart rate, systolic arterial blood pressure, plasma tumor necrosis factor (TNF) activity, interleukin (IL)-6 concentration and urine cys-LT to creatinine ratio were evaluated. The rectal temperature, plasma TNF activity and IL-6 concentrations were significantly higher and systolic arterial blood pressure and heart rate significantly lower after endotoxin infusion. Cats treated with zafirlukast had a significantly higher blood pressure at 4h (P=0.002) compared to placebo. Urine cys-LT to creatinine ratio was significantly greater in the cats treated with zafirlukast compared to placebo (P=0.02). Zafirlukast administration ameliorated the acute hypotensive response to endotoxin in cats, but failed to significantly alter rectal temperature, heart rate or production of TNF and IL-6., (Copyright © 2010 ISFM and AAFP. Published by Elsevier Ltd. All rights reserved.)

Thy-1 is a cell surface protein important in immunologic and neurologic processes, including T cell activation and proliferation, and neuronal outgrowth. In murine thymocytes, Thy-1 is downregulated in response to norepinephrine (NE) through posttranscriptional destabilization of its mRNA mediated by βAR/AC/cAMP/PKA signaling. In this study we investigated factors involved in NE/cAMP-mediated Thy-1 mRNA destabilization in S49 thymoma cells, and identified a region containing two copies of the AUUUA regulatory element (ARE), a motif commonly associated with mRNA decay, in the Thy-1 mRNA 3' UTR. Insertion of the Thy-1 ARE region into a reporter gene, resulted in cAMP induced destabilization of the reporter gene mRNA. RNA-protein binding studies revealed multiple Thy-1 ARE binding proteins, including AUF1, HuR, and TIAR. RNA silencing of HuR enhanced cAMP-mediated downregulation of Thy-1 mRNA, in contrast, silencing AUF1 had no effect. Immunoblotting revealed multiple proteins phosphorylated by PKA as a result of NE or cAMP signaling. These results reveal that the machinery of NE/cAMP modulation of Thy-1 mRNA decay involves a cAMP responsive ARE in its 3' UTR and multiple site specific ARE binding proteins. These findings add to our knowledge of Thy-1 mRNA regulation and provide insight into the regulation of ARE containing mRNAs, which impacts stress-related immunosuppression., (Published by Elsevier Inc.)

Cats with inflammatory bronchial disease are usually treated with glucocorticoid (GC) drugs to reduce airway inflammation. Inhalant GC delivery can preserve airway effects while systemic effects are minimized. An appropriate dosage regimen for inhaled GC in cats has not been investigated. A blinded, randomized, cross-over study design was used to investigate the ability of three different dosages of the inhalant GC fluticasone propionate delivered by metered dose inhaler to ameliorate eosinophilic airway inflammation in cats with experimentally induced allergic airway inflammation. Further, suppression of the hypothalamic-pituitary-adrenal axis (HPAA) at each dose was assessed. Fluticasone administered at dosages of 44, 110, or 220 microg q 12h reduced airway eosinophilia by 74%, 82%, or 81%, respectively (no difference). None of the dose regimens tested caused HPAA suppression. We conclude that a twice daily dosage of 44 microg fluticasone should be evaluated for the management of cats with naturally occurring inflammatory bronchial disease., (Copyright 2009 ESFM and AAFP. Published by Elsevier Ltd. All rights reserved.)

Mif2p is the budding-yeast orthologue of the mammalian centromere-binding protein CENP-C. We have mapped domains of Saccharomyces cerevisiae Mif2p and studied the phenotyptic consequences of their deletion. Using chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assays, we have further shown that Mif2p binds in the CDEIII region of the budding-yeast centromere, probably in close spatial association with Ndc10p. Moreover, ChIP experiments show that Mif2p recruits to yeast kinetochores a substantial subset of inner and outer kinetochore proteins, but not the Ndc80 or Spc105 complexes. We have determined the crystal structure of the C-terminal, dimerization domain of Mif2p. It has a "cupin" fold, extremely similar both in polypeptide chain conformation and in dimer geometry to the dimerization domain of a bacterial transcription factor. The Mif2p dimer seems to be part of an enhanceosome-like structure that nucleates kinetochore assembly in budding yeast.

Purpose: To evaluate the concordance between HER2 gene amplification, determined by fluorescence in situ hybridization (FISH), and HER2 protein overexpression assessed by an immunohistochemical (IHC) assay. The IHC protocol used was a research assay, known as the Clinical Trial Assay (CTA), developed to select women with metastatic breast cancer (MBC) for three pivotal clinical trials of trastuzumab therapy., Methods: A direct-labeled, dual-probe FISH assay was used to determine HER2 amplification in 623 fixed breast cancer tissue specimens. These specimens had been stored as paraffin-embedded sections for 25 years. All specimens had been analyzed for HER2 protein expression by the CTA. To assess the reproducibility of FISH results in archived material, we evaluated a separate group of 617 breast cancer tissue specimens at two different laboratories., Results: Informative FISH results were available for 529 (85%) of the 623 specimens. Overall concordance between FISH and IHC results was 82% (95% CI; 7885%). Assay agreement between FISH results and specimens with immunostaining scores of 0, 1+, and 3+ were 97, 93 and 89%, respectively. However, only 24% of specimens with 2+ immunostaining scores had HER2 amplification by FISH; there was assay disagreement in 76% of specimens in this IHC subgroup. Interlaboratory FISH concordance was 92% (95% CI; 8994%), indicating very good assay reproducibility in these archived specimens., Conclusion: HER2 status determined by CTA-IHC and FISH are significantly correlated; however, differences between these two assays can a ect patient selection for trastuzumab therapy.

Background: New strategies for cancer therapy include the combination of angiogenesis inhibitors with cytotoxins. However, angiogenesis inhibitors may alter tumor microvessel structure and transendothelial permeability thereby reducing tumoral delivery of cytotoxic agents. The aim of this study was to estimate quantitatively the apparent permeability-surface area product (K(PS)) in tumors to a macromolecular contrast medium (MMCM), to follow changes in K(PS) induced by antibodies to vascular endothelial growth factor (anti-VEGF), and to correlate the findings with tumor accumulation of cisplatin, a highly protein-bound cytotoxin, and 5-fluorouracil (5-FU), a small unbound cytotoxin., Methods: Dynamic MRI enhanced with a MMCM (albumin-(Gd-DTPA)(30)) was analyzed using a two-compartment tumor tissue model (plasma and interstitial water) to quantitatively estimate K(PS). These estimates of K(PS) were correlated with cytotoxic drug accumulations in the tumors., Results: Anti-VEGF treatment reduced K(PS) to MMCM in tumor tissue from 0.013 mL h(-1) cm(-3) (n = 9) at baseline to 0.003 mL h(-1) cm(-3) (n = 9) 24 h later (p <.05). The K(PS) values correlated significantly (r(2) =.78; p <.0001) with the tumor cisplatin accumulation. No correlation (r(2) =.001; p =.89) was found between K(PS) and tumor accumulation of the substantially smaller 5-FU molecule., Conclusions: MMCM-enhanced MRI can be used to detect and estimate changes in K(PS) to this contrast agent following a single dose of anti-VEGF antibody. The decline in K(PS) induced by this inhibitor of angiogenesis is associated with reduced tumor concentration of a protein-bound cytotoxin, similar in molecular weight to the contrast agent. MRI assays of microvascular status as performed here may be useful to clinically monitor responses to anti-angiogenesis drugs and to optimize the choice and timing of cytotoxic drug administration.

Skin extracellular matrix (ECM) molecules regulate a variety of cellular activities, including cell movement, which are central to wound healing and metastasis. Regulated cell movement is modulated by proteases and their associated molecules, including the serine proteases urinary-type plasminogen activator (uPA) and tissue-type plasminogen activator (tPA) and their inhibitors (PAIs). As a result of wounding and loss of basement membrane structure, epidermal keratinocytes can become exposed to collagen. To test the hypothesis that during wounding, exposed collagen, the most abundant ECM molecule in the skin, regulates keratinocyte PA and PAI gene expression, we utilized an in vitro model in which activated keratinocytes were cultured in dishes coated with collagen or other ECM substrates. tPA, uPA, and PAI-1 mRNA and enzymatic activity were detected when activated keratinocytes attached to fibronectin, vitronectin, collagen IV, and RGD peptide. In contrast, adhesion to collagen I and collagen III completely suppressed expression of PAI-1 mRNA and protein and further increased tPA expression and activity. Similarly, keratinocyte adhesion to laminin-1 suppressed PAI-1 mRNA and protein expression and increased tPA activity. The suppressive effect of collagen I on PAI-1 gene induction was dependent on the maintenance of its native fibrillar structure. Thus, it would appear that collagen- and laminin-regulated gene expression of molecules associated with plasminogen activation provides an additional dimension in the regulation of cell movement and matrix remodeling in skin wound healing.

A 67-yr-old woman with a 25-yr history of Crohn's disease, maintained on near-continuous corticosteroids (prednisone 10 mg daily) over a 6-yr period, underwent ileocolic resection for obstruction. Pathology revealed Crohn's disease, multiple nodules of Kaposi's sarcoma, and cytomegalic inclusion bodies with confirmation of cytomegalovirus by shell vial immunofluorescence. Testing for HIV serum antibody has been repeatedly negative. Crohn's disease, Kaposi's sarcoma, and cytomegalovirus have been clinically in remission for 5 yr.

Objectives: Neonatal thrombocytopenia occurs commonly in neonatal intensive care units. The role of the thrombopoietin (Tpo) system in normal neonatal platelet regulation and neonatal thrombocytopenia is not well understood. The purpose of our study was to: 1) determine the normal Tpo level at birth in healthy nonthrombocytopenic term (NTT) and nonthrombocytopenic preterm (NTP) infants and in infants born to women with preeclampsia; and 2) measure Tpo levels in infants during and after the resolution of thrombocytopenia. Characterizing Tpo levels in the healthy and thrombocytopenic newborn is an important step in furthering our understanding of the pathophysiology of neonatal thrombocytopenia., Methods: This study is comprised of 2 parts. For the first part, cord blood was obtained at birth from both term (gestational age [GA]: 38-42 weeks) and preterm (GA: 25-36 weeks) infants. If birth platelet levels were >/=140 x 10(3)/microL and the infant fit criteria for being normal, or if the infant was born to a women with preeclampsia, Tpo levels were measured. For the second part, serial Tpo levels and concomitant platelet counts (Plts) were measured in both preterm and term infants during a period of marked thromboctyopenia (Plt < 100 x 10(3)/microL) until its resolution (Plt >/= 140 x 10(3)/microL)., Results: Median cord blood Tpo levels from NTP infants (n = 35) were higher than those of NTT infants (n = 32; 95 pg/mL vs 48 pg/mL, respectively). In addition, preterm infants born to women with preeclampsia (n = 11) had lower Tpo levels than NTP infants with a similar GA (<41 pg/mL vs 95 pg/mL). For infants with marked thrombocytopenia, median Tpo levels during thrombocytopenia were similar between term (n = 12) and preterm (n = 14) groups (223 pg/mL and 179 pg/mL, respectively), with the majority of individuals showing a decrease in Tpo with resolution of thrombocytopenia. Within each group, there was large variability in the Tpo response to thrombocytopenia., Impression: These data show that the Tpo system is intact in NTP and NTT neonates. Preeclampsia may be an example of a disorder that perturbs this system. The great variability in Tpo levels seen in infants during thrombocytopenia may be related to the mechanism of thrombocytopenia. The finding that, in general, Tpo levels decreased with resolution of thrombocytopenia is consistent with what has been described in adults and children.

Starting out from our previous observations that defects in the immune system-brain feedback predispose to pathogenic immune responses, our interest focuses at the roles of adrenergic/cholinergic neurotransmitters in brain-immune interactions. We have shown in rodent models that 1) both catecholamines and acetylcholine are potent modulators of peripheral immune functions, 2) cholinergic signals are involved in the afferent signalling of the immune system, and 3) lymphocytes not only express functional adrenergic and cholinergic receptors, but synthesize and release neurotransmitters, such as acetylcholine, in quantitative dependence of differentiation and activation. Studies are presently being initiated to investigate the role(s) of these non-neuronal neurotransmitters within immune tissues, and to explore the relevance of excitatory amino acids as important central neurotransmitters in the brain-immune system dialogue.

Our work is devoted to defining relationships between the immune system and the adrenergic and cholinergic systems in vivo. In the rat model, we have shown that the cells of different immune compartments express the genes of a defined set of adrenergic/cholinergic receptors, and it was shown that lymphocytes are a site of non-neuronal production of norepinephrine and acetylcholine. Furthermore, using implantable slow-release tablets containing adrenergic or cholinergic agonists/antagonists, distinct and partly opposite effects were observed on peripheral immune functions. Concerning sympathetic immunoregulation, our data--in contrast to those of other studies--suggest that an enhanced adrenergic tonus leads to immunosuppression primarily via alpha 2-receptor-mediated mechanisms. Beta-blockade strongly enhances this effect, most likely by inhibition of pineal melatonin synthesis. In recent experiments on the kinetics it was found that the continuous alpha-adrenergic treatment entails a strong suppression of cellular responsiveness during the first few hours, which is increasingly followed by a general loss of lymphocytes in blood and lymphoid organs most likely due to enhanced apoptosis. More recently, we have extended our studies to the mouse model. First data obtained with RNAse protection assays suggest a biphasic effect on the gene expression of several cytokines in spleen cells due to adrenergic in vivo treatment.

We have previously shown that norepinephrine (NE) inhibits the in vitro generation of anti-MOPC-315 CTL activity by spleen cells from BALB/c mice rejecting a large MOPC-315 tumor as a consequence of low-dose melphalan (l -phenylalanine mustard (l -PAM)) treatment (l -PAM TuB spleen cells). Since TNF-alpha plays a key role in the generation of antitumor CTL activity in this system, we determined whether NE mediates this inhibition through inhibition of TNF-alpha production. Here, we show that NE inhibits the production of TNF-alpha protein and mRNA by l -PAM TuB spleen cells stimulated in vitro with mitomycin C-treated tumor cells. Flow cytometric analysis of intracellular expression of TNF-alpha revealed substantial NE-mediated decreases in the percentages of TNF-alpha+ cells among CD4+ and CD8+ T cells and F4/80+ activated macrophages. NE inhibition of CTL generation was largely overcome by addition of TNF-alpha to the stimulation cultures. When the beta-adrenergic antagonist propranolol was added to the stimulation cultures of l -PAM TuB spleen cells at a concentration that prevented NE-induced cAMP elevation, the NE-mediated decrease in TNF-alpha mRNA and NE-mediated inhibition of CTL generation were reversed. Collectively, these results suggest that NE inhibits antitumor CTL generation, at least in part, by inhibiting TNF-alpha synthesis through a mechanism(s) involving beta-adrenergic receptor signaling.

Fas ligand (FasL) is produced by activated T cells and natural killer cells and it induces apoptosis (programmed cell death) in target cells through the death receptor Fas/Apol/CD95. One important role of FasL and Fas is to mediate immune-cytotoxic killing of cells that are potentially harmful to the organism, such as virus-infected or tumour cells. Here we report the discovery of a soluble decoy receptor, termed decoy receptor 3 (DcR3), that binds to FasL and inhibits FasL-induced apoptosis. The DcR3 gene was amplified in about half of 35 primary lung and colon tumours studied, and DcR3 messenger RNA was expressed in malignant tissue. Thus, certain tumours may escape FasL-dependent immune-cytotoxic attack by expressing a decoy receptor that blocks FasL.