Approach to assess the economic impact of bovine tuberculosis in Ethiopia

AbstractBovine TB is prevalent in Ethiopian cattle and represents a serious zoonotic risk. However, extensive epidemiological data in the human and livestock sector are lacking. Create a dynamic transmission model of disease between animal and human, as a prerequisite for economic analysis of the most profitable intervention to control BTB in Ethiopia. Study on-going (2005-2010), epidemiological (prevalence, risk factors) and cost (human and livestock) data are collected in eight sites over a period of four years and fed into a compartmental trans-sectoral framework that simulates disease transmission. Different intervention scenarios will then be simulated in the model. The most profitable intervention to control BTB in Ethiopia has to be assessed as well as the cost sharing scheme between the public health and agricultural sectors. It has been postulated that a test and slaughter policy would have a negative economic impact in Ethiopia. Alternatives need to be assessed. [Ethiop.J.Health Dev. 2008;22(Special Issue):135-13

Effects of Hypoxia and Acidosis on Cardiac Electrophysiology and Hemodynamics. Is NHE-Inhibition by Cariporide Still Advantageous?

Hypoxia often leads to severe cardiac malfunctions. It is assumed that intracellular calcium overload is -inter alia- responsible for left ventricular (LV) deterioration. Inhibition of the sodium-proton exchanger (NHE), which finally inhibits/slows calcium overload, may ameliorate cardiac function. Our aim was to evaluate cariporide, an inhibitor of NHE1 in a Langendorff-perfused heart model. To discriminate a potentially different impact of extracellular acidosis and hypoxia we examined 48 Chinchilla Bastard rabbits divided into 8 experimental groups: control group (pH = 7.4, O2 = 100%) without or with cariporide (1µM), acidosis group (pH = 7.0, O2 = 100%) without or with cariporide (1µM), hypoxia group (pH = 7.4, O2 = 40%) without or with cariporide (1µM) and hypoxia+acidosis group (pH = 7.0, O2 = 40%) without or with cariporide (1µM). Hearts were subjected to acidotic/hypoxic conditions for 90 min followed by 60 min of reperfusion. Hypoxia and hypoxia+acidosis led to a severe deterioration of LV function with a decrease in LV pressure by about 70% and an increase of end-diastolic pressure from 6.7 ± 0.6 to 36.8 ± 5.4 (hypoxia) or from 7.0 ± 0.2 to 18.6 ± 4.1 (hypoxia+acidosis). Moreover, maximum contraction velocity decreased from about 1,800 mmHg/s to 600 mmHg/s during hypoxia ± acidosis and maximum relaxation velocity deteriorated from −1,500 mmHg/s to about −600 mmHg/s. During reperfusion hearts subjected to hypoxia+acidosis recovered faster than hearts subjected to hypoxia alone, reaching control levels after 5 min of reperfusion. Electrophysiologic analysis revealed an 1.2 fold increase in both dispersion of activation-recovery interval and in total activation time in the hypoxia ± acidosis group. Cariporide application significantly improved LV hemodynamics and electrophysiology in the hypoxia group but not in the group subjected to hypoxia+acidosis. Immunohistologic analysis of cardiac specimen revealed a significant increase of factors involved in hypoxia/reperfusion injury like nitrotyrosine and poly-ADP-ribose as well as apoptosis-inducing factors like AIF or cleaved-caspase 3 in LV after hypoxia ± acidosis. ATP was reduced by hypoxia but not by acidosis. Again, cariporide mitigated these processes only in the hypoxia alone group, but not in the group with additional acidosis. Acidosis without hypoxia only marginally disturbed LV function and electrophysiology, and was not affected by cariporide. Thus, our study demonstrated that several detrimental effects of hypoxia were mitigated or abrogated by acidosis and that NHE-inhibition improved only hypoxia-induced cardiac dysfunction

Growth and differentiation of primary and passaged equine bronchial epithelial cells under conventional and air-liquid-interface culture conditions

<p>Abstract</p> <p>Background</p> <p>Horses develop recurrent airway obstruction (RAO) that resembles human bronchial asthma. Differentiated primary equine bronchial epithelial cells (EBEC) in culture that closely mimic the airway cells <it>in vivo </it>would be useful to investigate the contribution of bronchial epithelium in inflammation of airway diseases. However, because isolation and characterization of EBEC cultures has been limited, we modified and optimized techniques of generating and culturing EBECs from healthy horses to mimic <it>in vivo </it>conditions.</p> <p>Results</p> <p>Large numbers of EBEC were obtained by trypsin digestion and successfully grown for up to 2 passages with or without serum. However, serum or ultroser G proved to be essential for EBEC differentiation on membrane inserts at ALI. A pseudo-stratified muco-ciliary epithelium with basal cells was observed at differentiation. Further, transepithelial resistance (TEER) was more consistent and higher in P₁cultures compared to P₀cultures while ciliation was delayed in P₁cultures.</p> <p>Conclusions</p> <p>This study provides an efficient method for obtaining a high-yield of EBECs and for generating highly differentiated cultures. These EBEC cultures can be used to study the formation of tight junction or to identify epithelial-derived inflammatory factors that contribute to lung diseases such as asthma.</p

Effects of autologous bone marrow stem cell transplantation on beta-adrenoceptor density and electrical activation pattern in a rabbit model of non-ischemic heart failure

BACKGROUND: Since only little is known on stem cell therapy in non-ischemic heart failure we wanted to know whether a long-term improvement of cardiac function in non-ischemic heart failure can be achieved by stem cell transplantation. METHODS: White male New Zealand rabbits were treated with doxorubicine (3 mg/kg/week; 6 weeks) to induce dilative non-ischemic cardiomyopathy. Thereafter, we obtained autologous bone marrow stem cells (BMSC) and injected 1.5–2.0 Mio cells in 1 ml medium by infiltrating the myocardium via a left anterolateral thoracotomy in comparison to sham-operated rabbits. 4 weeks later intracardiac contractility was determined in-vivo using a Millar catheter. Thereafter, the heart was excised and processed for radioligand binding assays to detect β(1)- and β(2)-adrenoceptor density. In addition, catecholamine plasma levels were determined via HPLC. In a subgroup we investigated cardiac electrophysiology by use of 256 channel mapping. RESULTS: In doxorubicine-treated animals β-adrenoceptor density was significantly down-regulated in left ventricle and septum, but not in right ventricle, thereby indicating a typical left ventricular heart failure. Sham-operated rabbits exhibited the same down-regulation. In contrast, BMSC transplantation led to significantly less β-adrenoceptor down-regulation in septum and left ventricle. Cardiac contractility was significantly decreased in heart failure and sham-operated rabbits, but was significantly higher in BMSC-transplanted hearts. Norepinephrine and epinephrine plasma levels were enhanced in heart failure and sham-operated animals, while these were not different from normal in BMSC-transplanted animals. Electrophysiological mapping revealed unaltered electrophysiology and did not show signs of arrhythmogeneity. CONCLUSION: BMSC transplantation improves sympathoadrenal dysregualtion in non-ischemic heart failure

Global, regional, and national burden of colorectal cancer and its risk factors, 1990–2019: a systematic analysis for the Global Burden of Disease Study 2019

Funding: F Carvalho and E Fernandes acknowledge support from Fundação para a Ciência e a Tecnologia, I.P. (FCT), in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy i4HB; FCT/MCTES through the project UIDB/50006/2020. J Conde acknowledges the European Research Council Starting Grant (ERC-StG-2019-848325). V M Costa acknowledges the grant SFRH/BHD/110001/2015, received by Portuguese national funds through Fundação para a Ciência e Tecnologia (FCT), IP, under the Norma Transitória DL57/2016/CP1334/CT0006.proofepub_ahead_of_prin

The global burden of adolescent and young adult cancer in 2019 : a systematic analysis for the Global Burden of Disease Study 2019

Background In estimating the global burden of cancer, adolescents and young adults with cancer are often overlooked, despite being a distinct subgroup with unique epidemiology, clinical care needs, and societal impact. Comprehensive estimates of the global cancer burden in adolescents and young adults (aged 15-39 years) are lacking. To address this gap, we analysed results from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019, with a focus on the outcome of disability-adjusted life-years (DALYs), to inform global cancer control measures in adolescents and young adults. Methods Using the GBD 2019 methodology, international mortality data were collected from vital registration systems, verbal autopsies, and population-based cancer registry inputs modelled with mortality-to-incidence ratios (MIRs). Incidence was computed with mortality estimates and corresponding MIRs. Prevalence estimates were calculated using modelled survival and multiplied by disability weights to obtain years lived with disability (YLDs). Years of life lost (YLLs) were calculated as age-specific cancer deaths multiplied by the standard life expectancy at the age of death. The main outcome was DALYs (the sum of YLLs and YLDs). Estimates were presented globally and by Socio-demographic Index (SDI) quintiles (countries ranked and divided into five equal SDI groups), and all estimates were presented with corresponding 95% uncertainty intervals (UIs). For this analysis, we used the age range of 15-39 years to define adolescents and young adults. Findings There were 1.19 million (95% UI 1.11-1.28) incident cancer cases and 396 000 (370 000-425 000) deaths due to cancer among people aged 15-39 years worldwide in 2019. The highest age-standardised incidence rates occurred in high SDI (59.6 [54.5-65.7] per 100 000 person-years) and high-middle SDI countries (53.2 [48.8-57.9] per 100 000 person-years), while the highest age-standardised mortality rates were in low-middle SDI (14.2 [12.9-15.6] per 100 000 person-years) and middle SDI (13.6 [12.6-14.8] per 100 000 person-years) countries. In 2019, adolescent and young adult cancers contributed 23.5 million (21.9-25.2) DALYs to the global burden of disease, of which 2.7% (1.9-3.6) came from YLDs and 97.3% (96.4-98.1) from YLLs. Cancer was the fourth leading cause of death and tenth leading cause of DALYs in adolescents and young adults globally. Interpretation Adolescent and young adult cancers contributed substantially to the overall adolescent and young adult disease burden globally in 2019. These results provide new insights into the distribution and magnitude of the adolescent and young adult cancer burden around the world. With notable differences observed across SDI settings, these estimates can inform global and country-level cancer control efforts. Copyright (C) 2021 The Author(s). Published by Elsevier Ltd.Peer reviewe

Global burden of 288 causes of death and life expectancy decomposition in 204 countries and territories and 811 subnational locations, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

BACKGROUND Regular, detailed reporting on population health by underlying cause of death is fundamental for public health decision making. Cause-specific estimates of mortality and the subsequent effects on life expectancy worldwide are valuable metrics to gauge progress in reducing mortality rates. These estimates are particularly important following large-scale mortality spikes, such as the COVID-19 pandemic. When systematically analysed, mortality rates and life expectancy allow comparisons of the consequences of causes of death globally and over time, providing a nuanced understanding of the effect of these causes on global populations. METHODS The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021 cause-of-death analysis estimated mortality and years of life lost (YLLs) from 288 causes of death by age-sex-location-year in 204 countries and territories and 811 subnational locations for each year from 1990 until 2021. The analysis used 56 604 data sources, including data from vital registration and verbal autopsy as well as surveys, censuses, surveillance systems, and cancer registries, among others. As with previous GBD rounds, cause-specific death rates for most causes were estimated using the Cause of Death Ensemble model-a modelling tool developed for GBD to assess the out-of-sample predictive validity of different statistical models and covariate permutations and combine those results to produce cause-specific mortality estimates-with alternative strategies adapted to model causes with insufficient data, substantial changes in reporting over the study period, or unusual epidemiology. YLLs were computed as the product of the number of deaths for each cause-age-sex-location-year and the standard life expectancy at each age. As part of the modelling process, uncertainty intervals (UIs) were generated using the 2·5th and 97·5th percentiles from a 1000-draw distribution for each metric. We decomposed life expectancy by cause of death, location, and year to show cause-specific effects on life expectancy from 1990 to 2021. We also used the coefficient of variation and the fraction of population affected by 90% of deaths to highlight concentrations of mortality. Findings are reported in counts and age-standardised rates. Methodological improvements for cause-of-death estimates in GBD 2021 include the expansion of under-5-years age group to include four new age groups, enhanced methods to account for stochastic variation of sparse data, and the inclusion of COVID-19 and other pandemic-related mortality-which includes excess mortality associated with the pandemic, excluding COVID-19, lower respiratory infections, measles, malaria, and pertussis. For this analysis, 199 new country-years of vital registration cause-of-death data, 5 country-years of surveillance data, 21 country-years of verbal autopsy data, and 94 country-years of other data types were added to those used in previous GBD rounds. FINDINGS The leading causes of age-standardised deaths globally were the same in 2019 as they were in 1990; in descending order, these were, ischaemic heart disease, stroke, chronic obstructive pulmonary disease, and lower respiratory infections. In 2021, however, COVID-19 replaced stroke as the second-leading age-standardised cause of death, with 94·0 deaths (95% UI 89·2-100·0) per 100 000 population. The COVID-19 pandemic shifted the rankings of the leading five causes, lowering stroke to the third-leading and chronic obstructive pulmonary disease to the fourth-leading position. In 2021, the highest age-standardised death rates from COVID-19 occurred in sub-Saharan Africa (271·0 deaths [250·1-290·7] per 100 000 population) and Latin America and the Caribbean (195·4 deaths [182·1-211·4] per 100 000 population). The lowest age-standardised death rates from COVID-19 were in the high-income super-region (48·1 deaths [47·4-48·8] per 100 000 population) and southeast Asia, east Asia, and Oceania (23·2 deaths [16·3-37·2] per 100 000 population). Globally, life expectancy steadily improved between 1990 and 2019 for 18 of the 22 investigated causes. Decomposition of global and regional life expectancy showed the positive effect that reductions in deaths from enteric infections, lower respiratory infections, stroke, and neonatal deaths, among others have contributed to improved survival over the study period. However, a net reduction of 1·6 years occurred in global life expectancy between 2019 and 2021, primarily due to increased death rates from COVID-19 and other pandemic-related mortality. Life expectancy was highly variable between super-regions over the study period, with southeast Asia, east Asia, and Oceania gaining 8·3 years (6·7-9·9) overall, while having the smallest reduction in life expectancy due to COVID-19 (0·4 years). The largest reduction in life expectancy due to COVID-19 occurred in Latin America and the Caribbean (3·6 years). Additionally, 53 of the 288 causes of death were highly concentrated in locations with less than 50% of the global population as of 2021, and these causes of death became progressively more concentrated since 1990, when only 44 causes showed this pattern. The concentration phenomenon is discussed heuristically with respect to enteric and lower respiratory infections, malaria, HIV/AIDS, neonatal disorders, tuberculosis, and measles. INTERPRETATION Long-standing gains in life expectancy and reductions in many of the leading causes of death have been disrupted by the COVID-19 pandemic, the adverse effects of which were spread unevenly among populations. Despite the pandemic, there has been continued progress in combatting several notable causes of death, leading to improved global life expectancy over the study period. Each of the seven GBD super-regions showed an overall improvement from 1990 and 2021, obscuring the negative effect in the years of the pandemic. Additionally, our findings regarding regional variation in causes of death driving increases in life expectancy hold clear policy utility. Analyses of shifting mortality trends reveal that several causes, once widespread globally, are now increasingly concentrated geographically. These changes in mortality concentration, alongside further investigation of changing risks, interventions, and relevant policy, present an important opportunity to deepen our understanding of mortality-reduction strategies. Examining patterns in mortality concentration might reveal areas where successful public health interventions have been implemented. Translating these successes to locations where certain causes of death remain entrenched can inform policies that work to improve life expectancy for people everywhere. FUNDING Bill & Melinda Gates Foundation

Charakterisierung beta-adrenerger Rezeptoren auf Pferdeleymphozyten und deren Regulation unter dem Einfluß von Clenbuterol und Dexamethason

6. ZUSAMMENFASSUNG Charakterisierung b-adrenerger Rezeptoren auf Pferdelymphozyten und deren Regulation unter dem Einfluß von Clenbuterol und Dexamethason Getu Abraham Institut für Pharmakologie, Pharmazie und Toxikologie, Veterinärmedizinische Fakultät, Universität Leipzig, Deutschland Juni, 2001 86 S., 205 Lit., 7 Tab., 23 Abb. Über Vorkommen und Eigenschaften von b-Adrenozeptoren auf Pferdelymphozyten und ihre pharmakologische Regulation liegen bisher keine Untersuchungen vor. Mit der vorliegenden Arbeit sollten b-adrenerge Rezeptoren an intakten Pferdelymphozyten in Bindungsstudien mit dem Radioliganden (-)-(125I)-Iodocyanopindolol (ICYP), einem potenten b2-selektiven Adrenozeptorenantagonisten, charakterisiert und deren funktionelle Ansprechbarkeit durch Messung des Isoprenalin-induzierten Anstiegs des intrazellulären cAMP-Gehaltes mittels Radioimmunoassay bestimmt werden. Die Bindung von ICYP an intakten Pferdelymphozyten war schnell, sättigbar (maximale Anzahl der Bindungsstellen 320 ± 20 ICYP Bindungsstellen/Zelle, Mittelwert±SEM, n = 12) ) und hochaffin (KD-Wert für ICYP 14,37 ± 1,66 pmol/l, Mittelwert±SEM, n = 12). Verdrängungsexperimente mit Agonisten und Antagonisten zeigten eine Affinität und Stereoselektivität, wie sie bei b-Adrenozeptoren zu erwarten waren. Der nicht radioaktiv markierte b2-selektive Adrenozeptorantagonist ICI 118,551 verdrängte den Radioliganden ICYP aus seiner Bindung mit mehr als 1500fach höherer Affinität als der nicht radioaktiv markierte b1-selektive Adrenozeptorantagonist CGP 20712A. In Pferdelymphozyten können somit mehr als 90 % der b-Adrenozeptoren dem b2-Subtyp zugeordnet werden. Ein Anteil von weniger als 10 % an b1-Adrenozeptoren kann allerdings nicht ausgeschlossen werden. Die Bindungsstellen waren stereospezifisch, da das biologisch aktive (-)-Propranolol die Bindung von ICYP mit 40fach höherer Affinität verdrängte als (+)-Propranolol. b-adrenerge Agonisten verdrängten ICYP aus seiner spezifischen Bindung in der Rangfolge ihrer pharmakologischen Wirkungstärke, die typisch für den b2-Subtyp des Adrenozeptors ist: (-)-Isoprenalin > (-)-Adrenalin > (-)-Noradrenalin. Eine ähnliche Rangfolge wurde auch für die Agonist-induzierte cAMP-Bildung in Lymphozyten festgestellt. Aufgrund der dargestellten Befunde kann die Schlußfolgerung gezogen werden, daß ICYP ein geeigneter Radioligand ist, um b-adrenerge Rezeptoren an Pferdelymphozyten zu identifizieren und zu charakterisieren. Die Bindung von ICYP an intakte Lymphozyten stellt somit ein geeignetes Modell dar, um wichtige Informationen über die Funktion und Regulation des b-adrenergen Systems beim Pferd sowohl unter physiologischen (pathophysiologischen) Bedingungen als auch über Arzneimittel-induzierte Veränderungen in vivo zu gewinnen. Ausgehend von den beschriebenen Grundlagenuntersuchungen wurden in einer weiteren Versuchsreihe bei zwölf klinisch gesunden Pferden der Einfluß von Clenbuterol und Dexamethason lymphozytäre b2-Adrenozeptoren, hinsichtlich ihrer Dichte, ihrer Affinität zum Radioliganden und ihrer cAMP-Bildung (als Maß für die funktionelle Ansprechbarkeit der b2-Adrenozeptoren bestimmt über Stimulation durch 10 µmol/l Isoprenalin) untersucht. Während der 12tägigen Clenbuterolbehandlung (2 Ž 0,8 µg/kg/Tag, i. v.) fielen die Anzahl der b2-Adrenozeptoren an Lymphozyten und der durch (-)-Isoprenalin-induzierte Anstieg des intrazellulären cAMP-Gehaltes signifikant (p 90 %) to the b2-adrenoceptor subtype. A minor ( (-)-adrenaline > (-)-noradrenaline, which is typical for a b2-subtype of adrenergic receptor; the same order of potency was obtained for agonist-induced stimulation of lymphocyte cyclic AMP content. Thus, it can be concluded that ICYP is a promising compound for the reliable determination of the binding characteristics of b-adrenoceptors of equine lymphocytes and besides that, this might give an insight in providing physiologically significant information about the regulation of the b-adrenergic receptor system. Based upon this study, we further investigated, in 12 healthy thoroughbred horses, the effects of the b2-agonist clenbuterol and the glucocorticoid dexamethasone on the lymphocyte b2-adrenenergic receptor density and affinity as well as on its responsiveness (assessed by lymphocyte cyclic AMP responses to 10 µmol/l (-)-isoprenaline). Clenbuterol (2Ž0.8µg/kg/d, i. v. for 12 days) decreased ICYP binding sites only 48 h after application by ~30-40 %; concomitantly, lymphocyte cAMP response to (-)-isoprenaline was significantly reduced (p < 0.05, n = 8). The values remained at these low levels throughout the following 10 days of treatment. After withdrawal of clenbuterol the density and responsiveness of b2-adrenoceptors gradually increased, reaching pre-drug levels after 4 days. Next the effects of dexamethasone on clenbuterol-induced desensitisation were studied. Administration of dexamethasone (1Ž0.1mg/kg/d, i. v. for five days) immediately after clenbuterol withdrawal accelerated b2-adrenergic receptor recovery (p < 0.05, n = 4): only 24 hours after administration dexamethasone restored the number of binding sites and cAMP response to (-)-isoprenaline to levels statistically indistinguishable from the values before starting clenbuterol treatment. Three days after dexamethasone administration the density of lymphocyte b2-adrenoceptors was further increased about two fold the pre-treatment values, and this increase declined gradually after dexamethasone withdrawal, reaching baseline values after 4 days. Furthermore, in groups simultaneously exposed to both drugs dexamethasone completely prevented clenbuterol-induced decrease in lymphocyte b2-adrenoceptor density and responsiveness. No significant change was observed in the dissociation constant and, thus, in the affinity of ICYP to b-adrenoceptors remained unchanged during or after treatment with either clenbuterol or dexamethasone. It is concluded that dexamethasone (glucocorticoids) can reverse and prevent clenbuterol-induced down-regulation of the lymphocyte b2-adrenoceptors and thus, a combined therapy with clenbuterol and dexamethasone may be potentially beneficial in COPD of horses

Charakterisierung beta-adrenerger Rezeptoren auf Pferdeleymphozyten und deren Regulation unter dem Einfluß von Clenbuterol und Dexamethason

6. ZUSAMMENFASSUNG Charakterisierung b-adrenerger Rezeptoren auf Pferdelymphozyten und deren Regulation unter dem Einfluß von Clenbuterol und Dexamethason Getu Abraham Institut für Pharmakologie, Pharmazie und Toxikologie, Veterinärmedizinische Fakultät, Universität Leipzig, Deutschland Juni, 2001 86 S., 205 Lit., 7 Tab., 23 Abb. Über Vorkommen und Eigenschaften von b-Adrenozeptoren auf Pferdelymphozyten und ihre pharmakologische Regulation liegen bisher keine Untersuchungen vor. Mit der vorliegenden Arbeit sollten b-adrenerge Rezeptoren an intakten Pferdelymphozyten in Bindungsstudien mit dem Radioliganden (-)-(125I)-Iodocyanopindolol (ICYP), einem potenten b2-selektiven Adrenozeptorenantagonisten, charakterisiert und deren funktionelle Ansprechbarkeit durch Messung des Isoprenalin-induzierten Anstiegs des intrazellulären cAMP-Gehaltes mittels Radioimmunoassay bestimmt werden. Die Bindung von ICYP an intakten Pferdelymphozyten war schnell, sättigbar (maximale Anzahl der Bindungsstellen 320 ± 20 ICYP Bindungsstellen/Zelle, Mittelwert±SEM, n = 12) ) und hochaffin (KD-Wert für ICYP 14,37 ± 1,66 pmol/l, Mittelwert±SEM, n = 12). Verdrängungsexperimente mit Agonisten und Antagonisten zeigten eine Affinität und Stereoselektivität, wie sie bei b-Adrenozeptoren zu erwarten waren. Der nicht radioaktiv markierte b2-selektive Adrenozeptorantagonist ICI 118,551 verdrängte den Radioliganden ICYP aus seiner Bindung mit mehr als 1500fach höherer Affinität als der nicht radioaktiv markierte b1-selektive Adrenozeptorantagonist CGP 20712A. In Pferdelymphozyten können somit mehr als 90 % der b-Adrenozeptoren dem b2-Subtyp zugeordnet werden. Ein Anteil von weniger als 10 % an b1-Adrenozeptoren kann allerdings nicht ausgeschlossen werden. Die Bindungsstellen waren stereospezifisch, da das biologisch aktive (-)-Propranolol die Bindung von ICYP mit 40fach höherer Affinität verdrängte als (+)-Propranolol. b-adrenerge Agonisten verdrängten ICYP aus seiner spezifischen Bindung in der Rangfolge ihrer pharmakologischen Wirkungstärke, die typisch für den b2-Subtyp des Adrenozeptors ist: (-)-Isoprenalin > (-)-Adrenalin > (-)-Noradrenalin. Eine ähnliche Rangfolge wurde auch für die Agonist-induzierte cAMP-Bildung in Lymphozyten festgestellt. Aufgrund der dargestellten Befunde kann die Schlußfolgerung gezogen werden, daß ICYP ein geeigneter Radioligand ist, um b-adrenerge Rezeptoren an Pferdelymphozyten zu identifizieren und zu charakterisieren. Die Bindung von ICYP an intakte Lymphozyten stellt somit ein geeignetes Modell dar, um wichtige Informationen über die Funktion und Regulation des b-adrenergen Systems beim Pferd sowohl unter physiologischen (pathophysiologischen) Bedingungen als auch über Arzneimittel-induzierte Veränderungen in vivo zu gewinnen. Ausgehend von den beschriebenen Grundlagenuntersuchungen wurden in einer weiteren Versuchsreihe bei zwölf klinisch gesunden Pferden der Einfluß von Clenbuterol und Dexamethason lymphozytäre b2-Adrenozeptoren, hinsichtlich ihrer Dichte, ihrer Affinität zum Radioliganden und ihrer cAMP-Bildung (als Maß für die funktionelle Ansprechbarkeit der b2-Adrenozeptoren bestimmt über Stimulation durch 10 µmol/l Isoprenalin) untersucht. Während der 12tägigen Clenbuterolbehandlung (2 Ž 0,8 µg/kg/Tag, i. v.) fielen die Anzahl der b2-Adrenozeptoren an Lymphozyten und der durch (-)-Isoprenalin-induzierte Anstieg des intrazellulären cAMP-Gehaltes signifikant (p 90 %) to the b2-adrenoceptor subtype. A minor ( (-)-adrenaline > (-)-noradrenaline, which is typical for a b2-subtype of adrenergic receptor; the same order of potency was obtained for agonist-induced stimulation of lymphocyte cyclic AMP content. Thus, it can be concluded that ICYP is a promising compound for the reliable determination of the binding characteristics of b-adrenoceptors of equine lymphocytes and besides that, this might give an insight in providing physiologically significant information about the regulation of the b-adrenergic receptor system. Based upon this study, we further investigated, in 12 healthy thoroughbred horses, the effects of the b2-agonist clenbuterol and the glucocorticoid dexamethasone on the lymphocyte b2-adrenenergic receptor density and affinity as well as on its responsiveness (assessed by lymphocyte cyclic AMP responses to 10 µmol/l (-)-isoprenaline). Clenbuterol (2Ž0.8µg/kg/d, i. v. for 12 days) decreased ICYP binding sites only 48 h after application by ~30-40 %; concomitantly, lymphocyte cAMP response to (-)-isoprenaline was significantly reduced (p < 0.05, n = 8). The values remained at these low levels throughout the following 10 days of treatment. After withdrawal of clenbuterol the density and responsiveness of b2-adrenoceptors gradually increased, reaching pre-drug levels after 4 days. Next the effects of dexamethasone on clenbuterol-induced desensitisation were studied. Administration of dexamethasone (1Ž0.1mg/kg/d, i. v. for five days) immediately after clenbuterol withdrawal accelerated b2-adrenergic receptor recovery (p < 0.05, n = 4): only 24 hours after administration dexamethasone restored the number of binding sites and cAMP response to (-)-isoprenaline to levels statistically indistinguishable from the values before starting clenbuterol treatment. Three days after dexamethasone administration the density of lymphocyte b2-adrenoceptors was further increased about two fold the pre-treatment values, and this increase declined gradually after dexamethasone withdrawal, reaching baseline values after 4 days. Furthermore, in groups simultaneously exposed to both drugs dexamethasone completely prevented clenbuterol-induced decrease in lymphocyte b2-adrenoceptor density and responsiveness. No significant change was observed in the dissociation constant and, thus, in the affinity of ICYP to b-adrenoceptors remained unchanged during or after treatment with either clenbuterol or dexamethasone. It is concluded that dexamethasone (glucocorticoids) can reverse and prevent clenbuterol-induced down-regulation of the lymphocyte b2-adrenoceptors and thus, a combined therapy with clenbuterol and dexamethasone may be potentially beneficial in COPD of horses