RESUMO
AIMS: Bioequivalence (BE) trials aim to demonstrate that the 90% confidence interval of the T/R-ratio of the pharmacokinetic metrics between two formulations (test [T] and reference [R]) of a drug is fully included in the acceptance interval [0.80, 1.25]. Traditionally, the sample size of BE trials is based on a power calculation based on the intrasubject variability coefficient of variation (CV) and the T/R-ratio of the metrics. Since the exact value of the T/R-ratio is not known prior to the trial, it is often assumed that the difference between the treatments does not exceed 5%. Hence, uncertainty about the T/R-ratio is expressed by using a fixed value for the sample size calculation. We propose to characterise the uncertainty about the T/R-ratio by a (normal) distribution for the log(T/R-ratio), with an assumed mean of log θ = 0.00 (i.e. θ = 1.00) and a standard deviation σu , which quantifies the uncertainty. Evaluating this distribution leads to the statistical assurance of the BE trial. METHODS: The assurance of a clinical trial can be derived by integrating the power over the distribution of the input parameters, in this case, the assumed distribution of the log(T/R)-ratio. Because it is an average power, the assurance can be interpreted as a measure of the probability of success that does not depend on a specific assumed value for the log(T/R)-ratio. The relationship between power and assurance will be analysed by comparing the numerical outcomes. RESULTS: Using the assurance concept, values of the standard deviation for the distribution of potential log(T/R)-ratios can be chosen to reflect the magnitude of uncertainty. For most practical cases (i.e. when 0.95 ≤ θ ≤ 1.05), the sample size is not, or only slightly, changed when σ = |log(θ)|. CONCLUSION: The advantage of deriving the assurance for BE trials is that uncertainty is directly expressed as a parameter of variability.
Assuntos
Ensaios Clínicos como Assunto/métodos , Modelos Estatísticos , Preparações Farmacêuticas/administração & dosagem , Projetos de Pesquisa , Humanos , Preparações Farmacêuticas/metabolismo , Farmacocinética , Probabilidade , Tamanho da Amostra , Equivalência Terapêutica , IncertezaRESUMO
INTRODUCTION: Currently, no universally accepted definition of extended half-life (EHL) recombinant FVIII (rFVIII) exists. Identifying the minimum half-life extension ratio required for a reduction in dosing frequency compared with standard rFVIII could enable a more practical approach to decisions around prophylaxis with EHL rFVIII. AIM: To identify the half-life extension ratio required to decrease rFVIII dosing frequency by at least 1 day while maintaining the proportion of patients with plasma rFVIII levels above 1 IU/dL and without increasing the total weekly dose. METHODS: A previously published population pharmacokinetic model for standard rFVIII was used to estimate the percentage of patients with factor VIII (FVIII) levels always >1 IU/dL using various benchmark regimens. Using modelling, dosing frequency was reduced while rFVIII half-life was extended until the percentage of patients with FVIII >1 IU/dL equalled that of the benchmark regimen. RESULTS: Benchmark 3×/wk dosing totalling 100 IU/kg/wk of rFVIII resulted in 56.6% of patients with FVIII levels always >1 IU/dL. With 2×/wk dosing, totalling 80 or 90 IU/kg/wk, half-life extensions required to maintain 56.6% of patients at FVIII levels >1 IU/dL were 1.30 and 1.26, respectively. A half-life extension ratio of 1.33 was required to change dosing from every 48 hours to every 72 hours (both at 105 IU/kg/wk) while maintaining 92.8% of patients with FVIII >1 IU/dL. CONCLUSION: Based on this investigation, EHL rFVIII products should have a minimum half-life extension ratio of 1.3 to provide a reduction in dosing frequency from 3× to 2×/wk compared with standard rFVIII products while maintaining the same minimum FVIII trough level.
Assuntos
Fator VIII/administração & dosagem , Fator VIII/farmacocinética , Modelos Biológicos , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/farmacocinética , Relação Dose-Resposta a Droga , Fator VIII/uso terapêutico , Meia-Vida , Hemofilia A/tratamento farmacológico , Humanos , Proteínas Recombinantes/uso terapêuticoRESUMO
OBJECTIVES: To use a mouse model to investigate the relationships among the components of the systemic robusticity hypothesis (SRH): voluntary exercise on wheels, spontaneous physical activity (SPA) in cages, growth hormones, and skeletal robusticity, especially cranial vault thickness (CVT). MATERIALS AND METHODS: Fifty female mice from lines artificially selected for high running (HR) and 50 from nonselected control (C) lines were housed in cages with (Active) or without wheels (Sedentary). Wheel running and SPA were monitored daily. The experiment began at 24-27 days of age and lasted 12 weeks. Food consumption was measured weekly. Mice were skeletonized and their interparietal, parietal, humerus, and femur were µCT scanned. Mean total thickness of the parietal and interparietal bones was determined, along with thickness of the cortical and diploe layers individually. Geometric cross-sectional indicators of strength were calculated for the long bones. Blood samples were assayed for IGF-1 and IGFBP-3. RESULTS: Physical activity differed significantly among groups, based both on linetype (C vs. HR) and activity (A vs. S). However, contrary to our predictions, the ratio of IGF-1 to IGFBP-3 was higher in C mice than in HR mice. Neither CVT nor postcranial robusticity was affected by linetype or activity, nor were most measures of CVT and postcranial robusticity significantly associated with one another. DISCUSSION: Our results fail to provide support for the systemic robusticity hypothesis, suggesting it is important to rethink the long-standing theory that increased CVT in Homo erectus reflects increased physical activity compared other hominin species.
Assuntos
Hormônio do Crescimento/sangue , Resistência Física/fisiologia , Corrida/fisiologia , Crânio/anatomia & histologia , Animais , Cruzamento , Feminino , Masculino , CamundongosRESUMO
The f 2 test is generally used for comparing dissolution profiles. In cases of high variability, the f 2 test is not applicable, and the Multivariate Statistical Distance (MSD) test is frequently proposed as an alternative by the FDA and EMA. The guidelines provide only general recommendations. MSD tests can be performed either on raw data with or without time as a variable or on parameters of models. In addition, data can be limited-as in the case of the f 2 test-to dissolutions of up to 85% or to all available data. In the context of the present paper, the recommended calculation included all raw dissolution data up to the first point greater than 85% as a variable-without the various times as parameters. The proposed MSD overcomes several drawbacks found in other methods.
Assuntos
Química Farmacêutica , Solubilidade , Análise Multivariada , Estados Unidos , United States Food and Drug AdministrationRESUMO
The in vitro-in vivo correlation (IVIVC) (Food and Drug Administration 1997) aims to predict performances in vivo of a pharmaceutical formulation based on its in vitro characteristics. It is a complex process that (i) incorporates in a gradual and incremental way a large amount of information and (ii) requires information from different properties (formulation, analytical, clinical) and associated dedicated treatments (statistics, modeling, simulation). These results in many studies that are initiated and integrated into the specifications (quality target product profile, QTPP). This latter defines the appropriate experimental designs (quality by design, QbD) (Food and Drug Administration 2011, 2012) whose main objectives are determination (i) of key factors of development and manufacturing (critical process parameters, CPPs) and (ii) of critical points of physicochemical nature relating to active ingredients (API) and critical quality attribute (CQA) which may have implications in terms of efficiency, safety, and inoffensiveness for the patient, due to their non-inclusion. These processes generate a very large amount of data that is necessary to structure. In this context, the storage of information in a database (DB) and the management of this database (database management system, DBMS) become an important issue for the management of projects and IVIVC and more generally for development of new pharmaceutical forms. This article describes the implementation of a prototype object-oriented database (OODB) considered as a tool, which is helpful for decision taking, responding in a structured and consistent way to the issues of project management of IVIVC (including bioequivalence and bioavailability) (Food and Drug Administration 2003) necessary for the implementation of QTPP.
Assuntos
Bases de Dados Factuais , Modelos Biológicos , Preparações Farmacêuticas/administração & dosagem , Disponibilidade Biológica , Química Farmacêutica/métodos , Desenho de Fármacos , Humanos , Preparações Farmacêuticas/química , Preparações Farmacêuticas/metabolismo , Projetos de Pesquisa , Equivalência Terapêutica , Estados Unidos , United States Food and Drug AdministrationRESUMO
BACKGROUND: Since doubts were raised, if a challenging medical procedure such as acute stroke treatment including thrombolysis with recombinant tissue plasminogen activator (rTPA) is available with identical standard and outcome 24 h and 7 days a week our aim was to examine if acute stroke patients defined by onset-admission time (OAT) of Assuntos
Isquemia Encefálica/tratamento farmacológico
, Isquemia Encefálica/epidemiologia
, Admissão do Paciente
, Acidente Vascular Cerebral/tratamento farmacológico
, Acidente Vascular Cerebral/epidemiologia
, Terapia Trombolítica
, Doença Aguda
, Idoso
, Idoso de 80 Anos ou mais
, Isquemia Encefálica/complicações
, Feminino
, Fibrinolíticos/uso terapêutico
, Humanos
, Hemorragias Intracranianas/complicações
, Hemorragias Intracranianas/tratamento farmacológico
, Hemorragias Intracranianas/epidemiologia
, Masculino
, Fotoperíodo
, Estudos Prospectivos
, Proteínas Recombinantes/uso terapêutico
, Sistema de Registros
, Índice de Gravidade de Doença
, Fatores Socioeconômicos
, Acidente Vascular Cerebral/complicações
, Fatores de Tempo
, Ativador de Plasminogênio Tecidual/uso terapêutico
, Resultado do Tratamento
RESUMO
AIM OF THE STUDY: For public health promotion purposes it is important to know how the general public perceives the risk factor "UV exposure" and how UV risk perception is connected to health-related attitudes and beliefs. The aim of the study was to collect representative data about UV risk perception in Germany. METHODS: A representative telephone survey using the ADM design was carried out among 1 501 German residents between May and July 2007. Variables related to UV risk knowledge, relevance of risk topics in every-day thinking and risk estimates were investigated. Data regarding a subjective benefit evaluation of UV exposition was also collected. RESULTS: The results suggest no essential gaps in the general knowledge about UV risks, except for the underestimation of UV-related cataracts. The respondents thought it extremely likely that UV exposure would cause health damage such as skin cancer, photoaging or sun burns. People were asked to report how often they had thought about a number of health risks including UV risks in the last two weeks. It was established that UV risks are present but not dominant in people's every-day thinking. Along with it, the risk evaluation proves to be rather moderate. The mean value for respondents' perceived personal risk is M=5.1, in the midsection of the given 10-point scale. The results show that perceived personal risk is not influenced by the serious UV health risks such as cancer or cataracts. The results also indicate that UV benefit and UV risk perception are not strongly related to one another. CONCLUSIONS: In summary, participants' perception and assessment of the various health risks of UV exposure seem to be realistic, but the UV risk assessment it is not related to the perception of personal risk. One must therefore assume that UV health risk information and education campaigns do not necessarily lead to a change in the perception of personal UV risk. Rather than addressing primarily risk knowledge, UV risk communication should focus more on motivational aspects and consider the role of UV benefit perception.
Assuntos
Atitude Frente a Saúde , Exposição Ambiental/efeitos adversos , Conhecimentos, Atitudes e Prática em Saúde , Opinião Pública , Lesões por Radiação/epidemiologia , Dermatopatias/epidemiologia , Raios Ultravioleta/efeitos adversos , Feminino , Alemanha , Humanos , Masculino , Vigilância da População , Prevalência , Lesões por Radiação/etiologia , Medição de Risco , Fatores de Risco , Dermatopatias/etiologia , Inquéritos e QuestionáriosRESUMO
OBJECTIVE: To assess bioequivalence between Equasym Retard and Medikinet retard containing 20 mg methylphenidate (MPH) hydrochloride in a fed state. MATERIALS: Equasym Retard 20 mg capsules (UCB, Monheim, Germany) and Medikinet retard 20 mg capsules (Medice, Iserlohn, Germany). METHODS: This was an open, single-center, randomized, 2-period, 2-sequence, balanced cross-over study with a wash-out period of 1 week between administrations in 14 healthy male and female volunteers, aged 18 - 45 years. Blood samples were collected over 24 hours and methylphenidate plasma concentration-time data were used to calculate pharmacokinetic metrics for both formulations. The main metrics were AUC0-t and Cmax. Bioequivalence was concluded if the 90% confidence interval (CI) for the ratio between test and reference was 80 - 125% (AUC0-t, Cmax). RESULTS: All dosed subjects finished both treatment periods and were included in pharmacokinetic and safety analyses. The adverse events observed, mainly nervous system disorders (headache), were all mild or moderate in intensity and resolved without any action taken. The adverse event profile was consistent with the currently applicable SmPCs (Summaries of Product Characteristics) for Equasym Retard and Medikinet retard. Geometric means +/- SD for AUC0-t and Cmax were 35.5 +/- 10.1 ng x h/ml and 4.05 +/- 0.96 ng/ml (Equasym Retard) and 39.2 +/- 13.8 ng x h/ml and 5.26 +/- 2.11 ng/ml (Medikinet retard). The 90% geometric confidence interval for AUC0-t (extent of absorption) was within limits accepted for bioequivalence. Bioequivalence could not be demonstrated for the rate of bioavailability (Cmax); both the lower confidence limit and the point estimate were below 80% of the reference. The study has shown that both formulations lead to a similar pattern of absorption and elimination following single dose administration in the fed state, although the test formulation shows a somewhat slimmer profile, where the first peak is less pronounced. No bioequivalence could be shown within the first 4 hours. The second peak of the test was also lower than the one of the reference (both lower confidence limit and point estimate below 80%). CONCLUSIONS: The two formulations are not bioequivalent, especially if the rate and values within the first four hours after administration are taken into account.
Assuntos
Estimulantes do Sistema Nervoso Central/administração & dosagem , Estimulantes do Sistema Nervoso Central/farmacocinética , Preparações de Ação Retardada/farmacocinética , Metilfenidato/administração & dosagem , Metilfenidato/farmacocinética , Adolescente , Adulto , Estimulantes do Sistema Nervoso Central/efeitos adversos , Preparações de Ação Retardada/efeitos adversos , Feminino , Humanos , Masculino , Metilfenidato/efeitos adversos , Pessoa de Meia-Idade , Equivalência TerapêuticaRESUMO
Research investigating risk perception suggests that not only the quantitative parameters used in technical risk assessment (i.e., frequency and severity of harm) but also 'qualitative' aspects such as the dread a hazard provokes or its controllability influence risk judgments. It remains to be elucidated, however, which neural mechanism underlie risk ratings in healthy subjects. Using fMRI to detect changes in neural activity we compared the neural activations elicited by risk ratings with those elicited by a letter detection task performed on the same stimuli. The latter task served to control for basic stimulus processing, response selection and button-pressing during task performance. Risk ratings differentially activated the medial prefrontal cortex, the inferior frontal gyrus, the cerebellum (P<0.05, FWE corrected, whole brain approach), and in an additional ROI analysis the amygdala (P<0.05, FWE corrected). Of these structures, particularly the amygdala and the prefrontal cortex have been previously associated with decisions about affective interference. Furthermore our data suggest both, similarities and differences between the neural correlates of risk ratings and risk taking as involved, for e.g., in gambling tasks.
Assuntos
Mapeamento Encefálico , Encéfalo/irrigação sanguínea , Tomada de Decisões/fisiologia , Imageamento por Ressonância Magnética , Assunção de Riscos , Adulto , Análise de Variância , Encéfalo/fisiologia , Humanos , Processamento de Imagem Assistida por Computador , Julgamento/fisiologia , Masculino , Oxigênio/sangueAssuntos
Fibrinolíticos/administração & dosagem , Acidente Vascular Cerebral/tratamento farmacológico , Ativador de Plasminogênio Tecidual/administração & dosagem , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Bases de Dados como Assunto , Feminino , Alemanha , Humanos , Infusões Intravenosas , Masculino , Pessoa de Meia-Idade , Seleção de Pacientes , Estudos Prospectivos , Proteínas Recombinantes/administração & dosagem , Resultado do TratamentoRESUMO
OBJECTIVE: To assess bioequivalence between an intact capsule and the content of a capsule sprinkled on applesauce. MATERIALS: Medikinet retard 20 mg capsules were obtained from Medice (Iserlohn, Germany). METHODS: This was a single-center, completely randomized, open, 2-period, 2-sequence, balanced crossover study with a washout period of 1 week between administrations, in 12 healthy male and female subjects, aged 18-45 years. Blood samples were collected over 24 hours and methylphenidate plasma concentration-time data were used to calculate pharmacokinetic parameters for both administrations. The main parameters were (confirmatory) AUC0-tz (extent of BA), Cmax, tmax (rate of BA) and (descriptively) AUC0-infinity and t1/2. Equivalence was concluded if the 90% confidence interval (CI) for the ratio between test and reference was 0.80-1.25 (AUC0-tz). RESULTS: All 12 dosed subjects finished both treatment periods and were included in pharmacokinetic and safety analyses. 90% geometric confidence intervals for AUC0-tz and Cmax data were well within accepted bioequivalence limits. The study has shown that both treatment modes lead to similar pattern of absorption and elimination following single-dose administration in the fed state. The test treatment (content of capsule sprinkled over 15 ml applesauce) is bioequivalent to the reference treatment (intact capsule) in terms of extent and rate of absorption. CONCLUSION: Data collected from this study demonstrate that Medikinet retard capsules can be opened and the content sprinkled on a tablespoon of applesauce without influencing the rate and extent of bioavailability.
Assuntos
Estimulantes do Sistema Nervoso Central/administração & dosagem , Estimulantes do Sistema Nervoso Central/farmacocinética , Alimentos , Metilfenidato/administração & dosagem , Metilfenidato/farmacocinética , Adolescente , Adulto , Área Sob a Curva , Cápsulas , Estudos Cross-Over , Preparações de Ação Retardada , Feminino , Meia-Vida , Humanos , Masculino , Equivalência TerapêuticaRESUMO
The specific absorption rate (SAR) is a prominent topic in the discussion about precautionary health protection. An experimental study investigated the effect of information about various SAR values (below the existing partial body limit value of 2 W/kg) on safety judgments of potential mobile phones users. It turns out that about 94% of the participants do not know the SAR value of their own mobile phone. SAR values below existing limits are not perceived as equally safe. Rather, the lower the SAR value, the higher the perceived safety. However, a majority of the participants does not consider these SAR values to be 100% safe, even if they are clearly below the existing limits. Explicitly indicating a precautionary limit value (referring to the Federal Office for Radiation Protection or to consumer organizations) does not change this safety evaluation. As expected, safety evaluation of the SAR values is also related to the perception of mobile phone risks. Those who are concerned about mobile phone communication give lower safety judgments than the unconcerned-independent of the level of the SAR values. Irrespective of that, our results suggest that establishing the SAR value as a criterion for mobile phones depends first of all on making it known to the public.
Assuntos
Atitude Frente a Saúde , Telefone Celular/estatística & dados numéricos , Qualidade de Produtos para o Consumidor , Exposição Ambiental/estatística & dados numéricos , Conhecimentos, Atitudes e Prática em Saúde , Proteção Radiológica/métodos , Medição de Risco/estatística & dados numéricos , Adulto , Carga Corporal (Radioterapia) , Tomada de Decisões , Feminino , Alemanha/epidemiologia , Humanos , Masculino , Doses de Radiação , Eficiência Biológica Relativa , Assunção de Riscos , Inquéritos e QuestionáriosRESUMO
Now-a-days immunoassays are world wide used for the rapid screening of drugs. Despite the fact that they are a highly valuable tool for the test of legal and illicit drugs, there is a non-negligible risk of false positive and false negative findings and many pitfalls must be taken into account when using these tests in an uncritical manner and without valid confirmation procedures.
Assuntos
Avaliação Pré-Clínica de Medicamentos/métodos , Imunoensaio/métodos , Reações Cruzadas , Reações Falso-Negativas , Reações Falso-Positivas , Humanos , Imunoensaio/efeitos adversos , Testes de Toxicidade/métodosRESUMO
Idiopathic hypertrophic chronic pachymeningitis (IHCP) is characterised by inflammatory fibrotic thickening of the dura mater. Long term management is controversial. A 28 year old man with craniospinal IHCP and prominent lymphocytic meningitis is reported. Cerebrospinal fluid and histological examination suggested a CD4+ T cell driven process and B cell stimulation. After surgical, tuberculostatic, and immunosuppressive treatment failed to control the progressive meningeal hypertrophy, causing severe headache and neurological disability, the disease process eventually abated with intraventricular cytarabine treatment.
Assuntos
Citarabina/uso terapêutico , Imunossupressores/uso terapêutico , Meningite/tratamento farmacológico , Adulto , Doença Crônica , Citarabina/administração & dosagem , Progressão da Doença , Dura-Máter/patologia , Cefaleia/etiologia , Humanos , Hipertrofia , Imunossupressores/administração & dosagem , Inflamação , Injeções Intraventriculares , Masculino , Meningite/patologia , Resultado do TratamentoRESUMO
RR01, a new highly lipophilic drug showing extremely low water solubility and poor oral bioavailability, has been incorporated into pH-dependent dissolving particles made of a poly(methacrylic acid-co-ethylacrylate) copolymer. The physicochemical properties of the particles were determined using laser-light-scattering techniques, scanning electron microscopy, high-performance liquid chromatography, and x-ray powder diffraction. Suspension of the free drug in a solution of hydroxypropylcellulose (reference formulation) and aqueous dispersions of pH-sensitive RR01-loaded nanoparticles or microparticles were administered orally to Beagle dogs according to a 2-block Latin square design (n = 6). Plasma samples were obtained over the course of 48 hours and analyzed by gas chromatography/mass spectrometry. The administration of the reference formulation resulted in a particularly high interindividual variability of pharmacokinetic parameters, with low exposure to compound RR01 (AUC0-48h of 6.5 microg x h/mL and coefficient of variation (CV) of 116%) and much higher Tmax, as compared to both pH-sensitive formulations. With respect to exposure and interindividual variability, nanoparticles were superior to microparticles (AUC0-48h of 27.1 microg x h/mL versus 17.7 microg x h/mL with CV of 19% and 40%, respectively), indicating that the particle size may play an important role in the absorption of compound RR01. The performance of pH-sensitive particles is attributed to their ability to release the drug selectively in the upper part of the intestine in a molecular or amorphous form. In conclusion, pH-dependent dissolving particles have a great potential as oral delivery systems for drugs with low water solubility and acceptable permeation properties.
Assuntos
Preparações Farmacêuticas/administração & dosagem , Resinas Acrílicas , Administração Oral , Animais , Cães , Portadores de Fármacos , Concentração de Íons de Hidrogênio , Absorção Intestinal , Masculino , Tamanho da Partícula , Preparações Farmacêuticas/química , Farmacocinética , Difração de Raios XRESUMO
Tacrine, a cholinesterase inhibitor for symptomatic treatment of minor to moderate dementia, and its primary metabolites 1-hydroxy-tacrine and 4-hydroxy-tacrine were studied by means of thin-layer chromatography, UV spectroscopy and gas-chromatography/mass spectroscopy. The analytical data (corrected hRf values, UV spectra in solution as well as reflectance spectra, high-pressure liquid chromatography data, GC retention indices and EI mass spectra) including various derivatization methods are described.
Assuntos
Inibidores da Colinesterase/farmacocinética , Tacrina/análogos & derivados , Tacrina/farmacocinética , Biotransformação , Inibidores da Colinesterase/efeitos adversos , Relação Dose-Resposta a Droga , Interações Medicamentosas , Humanos , Taxa de Depuração Metabólica , Relação Estrutura-Atividade , Tacrina/efeitos adversosRESUMO
Data are presented on a triplex type with two parallel homologous strands for which triplex formation is almost as strong as duplex formation at least for some sequences and even at pH 7 and 0.2 M NaCl. The evidence mainly rests upon comparing thermodynamic properties of similar systems. A paperclip oligonucleotide d(A12C4T12C4A12) with two linkers C4 obviously can form a triplex with parallel back-folded adenine strand regions, because the single melting transition of this complex splits in two transitions by introducing mismatches only in the third strand region. Respectively, a hairpin duplex d(A12C4T12) and a single strand d(A12) form a triplex as a 1:1 complex in which the second adenine strand is parallel oriented to the homologous one in the Watson-Crick paired duplex. In this system the melting temperature T(m) of the triplex is practically the same as that of the duplex d(A12)-d(T12), at least within a complex concentration range of 0.2-4.0 microM. The melting behaviour of complexes between triplex stabilizing ligand BePI and the system hairpin duplex plus single strand supports the triplex model. Non-denaturing gel electrophoresis suggests the existence of a triplex for a system in which five of the twelve A-T*A base triads are substituted by C-G*C base triads. The recognition between any substituted Watson-Crick base pair (X-Y) in the hairpin duplex d(A4XA7C4T7YT4) and the correspondingly replaced base (Z) in the third strand d(A4ZA7) is mutually selective. All triplexes with matching base substitutions (Z = X) have nearly the same stability (T(m) values from 29 to 33.5 degrees C), whereas triplexes with non-matching substitutions (Z not equal X) show a clearly reduced stability (T(m) values from 15 to 22 degrees C) at 2microM equimolar oligonucleotide concentration. Most nucleic acid triple helices hitherto known are limited to homopurine-homopyrimidine sequences in the target duplex. A stable triplex formation is demonstrated for inhomogeneous sequences tolerating at least 50% pyrimidine content in the homologous strands. On the basis of the surprisingly similar thermodynamic parameters for duplex and triplex, and of the fact that this triplex type seems to be more stable than many other natural DNA triplexes known, and on the basis of semiempirical and molecule mechanical calculations, we postulate bridging interactions of the third strand with the two other strands in the triplex according to the recombination motif. This triplex, denoted by us 'recombination-like form', tolerates heterogeneous base sequences.
Assuntos
DNA Recombinante/química , Pareamento de Bases , Temperatura Alta , Ligação de Hidrogênio , Cinética , Modelos Moleculares , Conformação de Ácido Nucleico , Desnaturação de Ácido Nucleico , Hibridização de Ácido Nucleico , Oligodesoxirribonucleotídeos/química , Análise Espectral/métodos , Relação Estrutura-Atividade , TermodinâmicaRESUMO
A fatality caused by ingestion of a decalcifying agent containing formic acid is reported. Quantitative analysis of formic acid in the form of its methyl ester was performed in different body fluids and organ samples using head-space gas chromatography with flame ionization detection. The blood taken at the time of admission to hospital had a concentration of 370.3 microg/ml, which declined to 13.9 microg/ml after 6.5 h of haemodialysis. Post-mortem concentrations were 855.4 microg/ml (heart blood), 2,712 microg/ml (gastric contents), 1128 microg/ml (haemorrhagic fluid from abdominal cavity), 3,051 microg/ml (bile), 2,664 microg/ml (contents of small intestine), 442.7 microg/g (liver) and 542.3 microg/g (kidney). The most important morphological findings for differentiating between oral and respiratory ingestion were ulceration of the oropharynx and the oesophagus as well as extensive necrotic lesions in the stomach and the duodenum without perforation. Death was caused by massive acidosis, haemolysis, bleeding complications, hepatic and renal failure. Toxicological and morphological findings revealed that a considerable amount of formic acid had been ingested orally with a suicidal intention.
Assuntos
Autopsia/métodos , Causas de Morte , Formiatos/intoxicação , Suicídio , Administração Oral , Adulto , Sistema Digestório/patologia , Ionização de Chama/métodos , Formiatos/metabolismo , Humanos , Exposição por Inalação , Masculino , Exposição Ocupacional/análise , Intoxicação/patologiaRESUMO
Two cases (involving five murder victims) of multiple homicide by inhalational chloroform intoxication are reported. In the discussion of the findings the valence of toxicological analyses is underlined with regard to the possibility of forcible external suffocation due to occlusion of the respiratory orifices by means of a chloroform-soaked soft covering. In addition storage experiments were performed at +4, +20 and -20 degrees C with cadaver blood mixed with chloroform. The optimal solution for avoiding volatile losses was stored in glass tubes with ground glass stoppers. In cases of unclear death in which involvement of volatile substances is suspected it is, therefore, advisable to preserve an additional blood sample at -20 degrees C in glass tubes that are only opened for the analysis of volatile substances.