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1.
Electrophoresis ; 38(13-14): 1771-1779, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28401988

RESUMO

Telogen hairs presented in the crime scene are commonly encountered as trace evidence. However, short tandem repeat (STR) profiling of the hairs currently have low and limited use due to poor success rate. To increase the success rate of STR profiling of telogen hairs, we developed a rapid and cost-effective method to estimate the number of nuclei in the hair roots. Five cationic dyes, Methyl green (MG), Harris hematoxylin (HH), Methylene blue (MB), Toluidine blue (TB), and Safranin O (SO) were evaluated in this study. We conducted a screening test based on microscopy and the percentage of loss with nuclear DNA, in order to select the best dye. MG was selected based on its specific nuclei staining and low adverse effect on the hair-associated nuclear DNA. We examined 330 scalp and 100 pubic telogen hairs with MG. Stained hairs were classified into five groups and analyzed by STR. The fast staining method revealed 70% (head hair) and 33.4% (pubic hair) of full (30 alleles) and high partial (18-29 alleles) STR profiling proportion from the lowest nuclei count group (one to ten nuclei). The results of this study demonstrated a rapid, specific, nondestructive, and high yield DNA profiling method applicable for screening telogen hairs.


Assuntos
Corantes/análise , Impressões Digitais de DNA/métodos , Cabelo/química , Cátions , Corantes/química , Eletroforese Capilar/métodos , Humanos , Verde de Metila/análise , Verde de Metila/química , Repetições de Microssatélites , Reação em Cadeia da Polimerase
2.
J Forensic Sci ; 62(4): 1080-1088, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28097647

RESUMO

Pathological diagnosis of drowning remains a challenge for forensic science, because of a lack of pathognomonic findings. We analyzed microbiota and surfactant protein in the lungs for a novel diagnosis of drowning. All rats were divided into drowning, postmortem submersion, and control groups. The water, lungs, closed organs (kidney and liver), and cardiac blood in rats were assayed by targeting 16S ribosomal RNA of Miseq sequencing. Lung samples were analyzed by immunohistochemical staining for surfactant protein A. The closed organs and cardiac blood of drowned group have a lot of aquatic microbes, which have not been detected in postmortem submersion group. Furthermore, intra-alveolar granular staining of surfactant protein A (SP-A) was severely observed in the drowned group than the postmortem submersion and control groups. The findings suggested that the presence of aquatic microbiota in the closed organs and increased expression of SP-A could be markers for a diagnosis of drowning.


Assuntos
Sangue/microbiologia , Afogamento/diagnóstico , Rim/microbiologia , Fígado/microbiologia , Pulmão/química , Proteína A Associada a Surfactante Pulmonar/análise , Animais , Biomarcadores/análise , Análise Química do Sangue , DNA Bacteriano/genética , Patologia Legal , Água Doce/microbiologia , Sequenciamento de Nucleotídeos em Larga Escala , Imuno-Histoquímica , Rim/química , Fígado/química , Pulmão/patologia , Microbiota , Modelos Animais , Reação em Cadeia da Polimerase , RNA Ribossômico 16S , Ratos Sprague-Dawley , Água do Mar/microbiologia
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