RESUMO
Although there are several in vivo tests for potential genotoxicity, with the possible exception of the transgenic rodent mutation models, none is specifically intended to assess increasing damage with chronic administration. In principle, peripheral blood lymphocytes would be expected to accumulate DNA damage with repeated dosing because the majority are not in active division and appear to have limited DNA repair capability, and they are exposed to plasma levels of test materials and metabolites. However, there appear to be no published reports confirming this principle. Therefore, in the current study, after optimising culture conditions for rat lymphocytes in this laboratory, rats were given oral doses of cyclophosphamide or hexamethylphosphoramide (HMPA) for up to 28 days and peripheral lymphocytes analysed for chromosome aberrations at various time points. The results clearly show that, for both compounds, doses that gave no significant increases in aberration frequency after 2 days induced clear increases after 15 days with further damage detectable after 28 doses. With HMPA, it was shown that DNA damage persisted for at least 10 days after cessation of treatment. These data show that repeat dose studies in the rat measuring chromosome aberration frequency in lymphocytes can give a genuine indication that genotoxicity may increase with chronic administration and, therefore, maybe useful in assessing the risk of potentially genotoxic substances.
Assuntos
Aberrações Cromossômicas/induzido quimicamente , Ciclofosfamida/toxicidade , Hempa/toxicidade , Linfócitos/efeitos dos fármacos , Mutagênicos/toxicidade , Animais , Células Cultivadas , Ciclofosfamida/administração & dosagem , Ciclofosfamida/farmacologia , Hempa/administração & dosagem , Hempa/farmacologia , Linfócitos/metabolismo , Masculino , Testes de Mutagenicidade , Mutagênicos/administração & dosagem , Mutagênicos/farmacologia , RatosRESUMO
Heparin-induced thrombocytopenia (HIT, type II) is an immune-mediated disorder due to antibodies formed against heparin-platelet factor 4 complexes, usually appearing at days 5 to 14 after initiation of heparin. It is important to recognize HIT because heparin prophylaxis or treatment paradoxically associates with new venous and/or arterial thrombosis. Early clinical suspicion and diagnosis together with proper pharmacotherapy and close laboratory monitoring are the cornerstones for successful management. This includes monitoring of Thrombocytopenia, its Timing to heparin administration, appearance of new Thrombosis or resistance to treatment, and differential diagnosis by exclusion of o Ther causes (the 4T's). Specific attention should be paid to the absence or presence of thrombosis and to tailoring thromboprophylaxis or anticoagulant therapy with a nonheparin alternative. Even in the absence of HIT-associated thrombosis, an active policy for prolonged thromboprophylaxis is demanded. Rapid and reliable assays should be developed for diagnosis and anticoagulation monitoring to secure safe management with nonheparins. Semiquantitative testing for on-call hours should be available and later confirmed as clinically needed. Alternative therapeutic options are available, but because their use is infrequent, experienced coagulation treatment centers should provide guidance in the treatment and in laboratory monitoring. Most of the evidence in HIT is grade IC, and thus the best evidence is provided by clinical experience. New anticoagulants and platelet inhibitors may offer future alternatives in the management of HIT, but the current treatment options provide the best experience and benefit. The joint clinical and laboratory guidelines provided in this article along with two practical case scenarios were prepared by a Nordic expert panel. They will be valuable for hematologists and colleagues who do not routinely encounter HIT.
Assuntos
Anticoagulantes/uso terapêutico , Heparina/efeitos adversos , Trombocitopenia/induzido quimicamente , Idoso , Arginina/análogos & derivados , Cateterismo Cardíaco/métodos , Procedimentos Cirúrgicos Cardíacos/métodos , Sulfatos de Condroitina/uso terapêutico , Dermatan Sulfato/uso terapêutico , Feminino , Heparina de Baixo Peso Molecular/uso terapêutico , Heparitina Sulfato/uso terapêutico , Hirudinas , Humanos , Masculino , Fragmentos de Peptídeos/uso terapêutico , Ácidos Pipecólicos/uso terapêutico , Proteínas Recombinantes/uso terapêutico , Sulfonamidas , Trombocitopenia/diagnóstico , Trombocitopenia/tratamento farmacológico , Procedimentos Cirúrgicos Vasculares/métodos , Vitamina K/antagonistas & inibidores , Adulto JovemRESUMO
BACKGROUND: The amount of method development and assay validation required to support analysis of solutions from in vitro systems is a consideration of analytical laboratories performing this type of analysis. As there is little information from regulatory bodies as to how much assay development and validation is required, analytical laboratories need to decide on the best approach for this type of work. In this paper, we describe an efficient 'fit-for-purpose' approach that has been developed to support buffer sample analysis from Safety Pharmacology hERG studies. RESULTS: Method development has been minimized with the aid of compound modeling software and generic HPLC-MS/MS analytical systems. The assay is evaluated prior to sample analysis using simple qualification procedures to support 'one-off' analyses. CONCLUSION: The result is an efficient process that allows speedy and confident analysis of in vitro samples to successfully support regulatory hERG in vitro studies without the additional workload of a full validation procedure.