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1.
Forensic Sci Int Genet ; 14: 187-90, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25450791

RESUMO

Recently there has been a drive towards standardisation of forensic DNA interpretation methods resulting in the uptake of probabilistic interpretation software. Some of these software solutions utilise Markov chain Monte Carlo techniques (MCMC). They will not produce an identical answer after repeat interpretations of the same evidence profile because of the Monte Carlo aspect. This is a new source of variability within the forensic DNA analysis process. In this paper we explore the size of the MCMC variability within the interpretation software STRmix™ compared to other sources of variability in forensic DNA profiling including PCR, capillary electrophoresis load and injection, and the makeup of allele frequency databases. The MCMC variability within STRmix™ was shown to be the smallest source of variability in this process.


Assuntos
Funções Verossimilhança , DNA/genética , Genética Forense , Humanos , Cadeias de Markov , Método de Monte Carlo
2.
Forensic Sci Int Genet ; 8(1): 20-3, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24315584

RESUMO

The forensic analysis of DNA is most often undertaken by the amplification of short tandem repeats (STR) using the polymerase chain reaction (PCR). DNA amplification can result in production of the target allele amplicon and a by-product called stutter. Stutter is the result of the miscopy of the target allele and is typically one repeat smaller. Stutter is traditionally described as a ratio of stutter and allele height; stutter ratio (SR). The challenge to DNA profile interpretation is most serious whenever stutter products are of a similar height to the minor allelic peaks in a mixed DNA profile. An accurate assignment of peaks and the prediction of their height is important when objectively interpreting forensic DNA profiles. The longest uninterrupted stretch (LUS) of tandem repeats within the allele has previously been shown to be a good predictor of stutter ratio. LUS is determined by sequencing a range of observed alleles at a locus. The locus D6S1043 is a relatively new locus to appear in commercial forensic DNA testing kits. To date however, there has been no comprehensive report of sequencing of this locus. In this work, we sequence a sample of D6S1043 alleles to determine LUS values and investigate allele repeat number and LUS as explanatory variables for SR.


Assuntos
Repetições de Microssatélites/genética , Alelos , Sequência de Bases , Primers do DNA , Heterozigoto , Humanos , Reação em Cadeia da Polimerase
3.
Neurosci Lett ; 445(1): 126-9, 2008 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-18775475

RESUMO

Isolated adrenal medullary chromaffin cells maintained in culture have been widely used to study neurosecretory events. Many of these studies have been conducted using cells obtained from the bovine adrenal. In this study we have cultured chromaffin cells from an alternative large animal model, the deer, and have conducted the first characterization of secretion from this preparation. Cervine chromaffin cells, preloaded with [3H]noradrenalin, displayed a strong secretory response to the cholinergic agonist carbachol, with a maximal secretion of approximately 28% cell content over 15 min. This response was reproduced by nicotinic but not muscarinic agonists and was similarly inhibited by nicotinic but not muscarinic antagonists. Nicotine-evoked secretion measured over a 15 min time period was inhibited approximately 50% by the L-type Ca2+-channel antagonist nifedipine and approximately 20% by N-type (omega-conotoxin GVIA) or N, P/Q-type (omega-conotoxin MVIIC) antagonists. In contrast the response was unaffected by omega-agatoxin IVA, a P/Q-type antagonist. In addition to nicotinic receptor stimulation, activation of PACAP or histamine H1 receptors resulted in a concentration-dependent increase in secretion. PACAP was approximately two-fold more effective than histamine although both were weaker secretagogues than nicotine. In contrast, cervine chromaffin cells did not respond to angiotensin II or bradykinin, two agents known to stimulate secretion from bovine chromaffin cells. These data provide an initial characterization of the secretory response from cervine adrenal medullary chromaffin cells indicating that there are marked similarities but also potentially significant differences between them and their far more extensively described bovine counterparts.


Assuntos
Catecolaminas/metabolismo , Células Cromafins/metabolismo , Cervos/anatomia & histologia , Glândulas Suprarrenais/citologia , Animais , Bloqueadores dos Canais de Cálcio/farmacologia , Carbacol/farmacologia , Células Cultivadas , Agonistas Colinérgicos/farmacologia , Células Cromafins/efeitos dos fármacos , Relação Dose-Resposta a Droga , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , Norepinefrina/metabolismo , Trítio/metabolismo
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