Brain tissue identification based on myosin heavy chain isoforms

Kimura, Akihiko; Ikeda, Hiroaki; Yasuda, Seiji; Yamaguchi, Kazuya; Tsuji, Tsutomu

doi:10.1007/bf01428404

Cited by 11 publications

(7 citation statements)

References 13 publications

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“…However, putrefied and heated specimens were not examined. According to our results, the present ELISA method was applicable to the dried brains maintained at 37 8C for at least 4 months, and this result was superior to past reports [11,12]. In addition, our ELISA method was useful for the putrefied and heated brains.…”

Section: Discussionsupporting

confidence: 48%

“…From this viewpoint, we attempted to detect human NF-H from experimentally putrefied, dried or heated human brains. In previously reported methods for identifying human brain, in which myosin or S-100 protein were quantitatively determined, the authors investigated the usefulness of their methods with dried specimens [11,12]. However, putrefied and heated specimens were not examined.…”

Section: Discussionmentioning

confidence: 99%

“…Although it occurs not so often, we reported some practical cases in which identification of human muscle or skin had been required [9,10]. Identification of the human brain is also important, especially in the traffic accidents such as hitand-run cases and some immunological methods for this purpose have previously been reported [11,12], however, they required quantitative analysis by measurement of fluorescence intensity. In this study, we attempted to develop a simpler and more qualitative method for identifying human brain by detection of human NF using an enzyme-linked immunosorbent assay (ELISA), and its usefulness was evaluated.…”

Section: Introductionmentioning

confidence: 91%

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Identification of human brain from a tissue fragment by detection of neurofilament proteins

Takata

Miyaishi

Kitao

et al. 2004

Forensic Science International

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We developed a method for identifying human brain from a tissue-like fragment by detection of neurofilament protein (NF) using enzyme-linked immunosorbent assay (ELISA). NF was extracted from 0.1 g of organ/tissue homogenized with Tris-HCl buffer (pH 7.2) containing urea, phenylmethylsulfonyl fluoride (PMSF), EDTA and, EGTA. It was necessary to dilute the extract at more than 2(3)-fold to avoid immunosuppression by urea. Positive reaction was always obtained for NF-H in 2(3)-fold diluted extract of brain tissue, however, NF-L and NF-M were not always detected when a brain fragment contained gray matter. Human cerebral white matter could be easily distinguished from other organs/tissues by detecting any of the NF-subunits. Brains of human and some animals could be discriminated by detecting NF-L or NF-M, although the species specificity of NF-H was not good. Our findings suggested that detection of NF-H was more useful than NF-L and NF-M for identifying a brain from a tissue-like fragment. The present ELISA method for NF-H could identify human brain specimens under the following conditions: putrefied at 4 degrees C for up to 3 weeks, dried at 37 degrees C for at least 4 months, heated at 50 degrees C for at least 4 weeks. Our results showed that our method is useful for identification of brain tissue in forensic stain analysis. Two practical cases are described.

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Section: Discussionsupporting

confidence: 48%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 91%

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Identification of human brain from a tissue fragment by detection of neurofilament proteins

Takata

Miyaishi

Kitao

et al. 2004

Forensic Science International

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“…Messenger RNAs transcribed tissue-specifically and can be utilized for tissue identification, however, it seems to be impractical at present because of their instability, suggesting that antibody-based detection is an indispensable tool for forensic investigations [1,2]. Messenger RNAs transcribed tissue-specifically and can be utilized for tissue identification, however, it seems to be impractical at present because of their instability, suggesting that antibody-based detection is an indispensable tool for forensic investigations [1,2].…”

Section: Introductionmentioning

confidence: 99%

Production and characterization of a monoclonal antibody for sweat-specific protein and its application for sweat identification

et al. 2003

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Identification of body fluids is a common task in medico-legal practice, but specific markers for sweat have not been identified to date. To develop a method for identification of sweat, we identified a sweat-specific protein and produced monoclonal antibodies by immunizing mice with sweat proteins fractionated by anion-exchange chromatography. Among many sweat-reactive monoclonal antibodies obtained, one monoclonal antibody (G-81) was selected because of its unique specificity. G-81 reacted to sweat but not to other body fluids (e.g. serum, saliva, semen, milk, urine and tears) in ELISA. G-81 specifically stained the eccrine sweat gland and did not stain any other tissue including the apocrine sweat gland. In western blotting, G-81 reacted strongly to a 7 kDa band and faintly to 20, 27 and 33 kDa bands of sweat protein. The N-terminal amino acid sequence (18 amino acids) of G-81-reactive peptides was determined, and an identical sequence was found in an antibiotic peptide dermcidin (110 amino acids) reported recently, suggesting that G-81 recognized a fragment of dermcidin. The G-81-reactive peptide could be detected in 8,192-fold dilutions of sweat by ELISA and could be detected in 200-fold diluted sweat samples (n=26) independent of the protein concentration. The G-81-reactive peptide was very stable and was able to detect sweat stains left for at least 11 weeks at room temperature without substantial loss of reactivity. These facts suggest that G-81 is a very useful tool for sweat identification in medico-legal practice.

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“…More recently, an mRNA assay was developed which allows for the inference of seven organ tissue types . Previously, enzyme‐linked immunosorbent assays or histological methods were used to infer the type of cell . These methods are, however, often not of sufficient sensitivity to assess the limited amount of cells present on forensic evidentiary traces .…”

Section: Introductionmentioning

confidence: 99%

Extended specificity studies of mRNA assays used to infer human organ tissues and body fluids

Berge

Sijen

2017

Electrophoresis

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Messenger RNA (mRNA) profiling is a technique increasingly applied for the forensic identification of body fluids and skin. More recently, an mRNA-based organ typing assay was developed which allows for the inference of brain, lung, liver, skeletal muscle, heart, kidney, and skin tissue. When applying this organ typing system in forensic casework for the presence of animal, rather than human, tissue is an alternative scenario to be proposed, for instance that bullets carry cell material from a hunting event. Even though mRNA profiling systems are commonly in silico designed to be primate specific, physical testing against other animal species is generally limited. In this study, human specificity of the organ tissue inferring system was assessed against organ tissue RNAs of various animals. Results confirm human specificity of the system, especially when utilizing interpretation rules considering multiple markers per cell type. Besides, we cross-tested our organ and body fluid mRNA assays against the target types covered by the other assay. Marker expression in the nontarget organ tissues and body fluids was observed to a limited extent, which emphasizes the importance of involving the case-specific context of the forensic samples in deciding which mRNA profiling assay to use and when for interpreting results.

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Brain tissue identification based on myosin heavy chain isoforms

Cited by 11 publications

References 13 publications

Identification of human brain from a tissue fragment by detection of neurofilament proteins

Identification of human brain from a tissue fragment by detection of neurofilament proteins

Production and characterization of a monoclonal antibody for sweat-specific protein and its application for sweat identification

Extended specificity studies of mRNA assays used to infer human organ tissues and body fluids

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