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Journal of Forensic Sciences | 1999

Simultaneous detection of multiple STR loci on sex chromosomes for forensic testing of sex and identity

Zaw Tun; Katsuya Honda; Masato Nakatome; Masami Nakamura; Shigeko Shimada; Yoshiaki Ogura; Hisanaga Kuroki; Motohiko Yamazaki; Masaru Terada; Ryoji Matoba

The forensic usefulness of X and Y chromosomal STR loci has recently been demonstrated. One quadruplex-PCR, using 2 X- and 2 Y-STRs (STRX1/HPRTB and DYS390/ DYS393), and 2 duplex-PCRs, each using an X- and a Y-STR (ARA/DYS390 and ARA/DYS393), and detection of PCR products by using an automated DNA sequencer are reported herein. This approach allows us to determine not only the sex of the donor of a sample, but also the X- and/or Y-STR genotypes of the sample. A male biological specimen yields 4 amplified products in quadruplex-PCR and 2 amplified fragments in duplex-PCRs, whereas a female biological specimen yields only 2 amplified fragments of X-STR in quadruplex-PCR and one fragment, also of X-STR, in duplex-PCRs. Our study thus provides useful information for many activities in forensic practice, such as identity testing, paternity testing, especially of deficiency cases, compilation of population data, and sex determination of a biological sample from a single PCR.


International Journal of Legal Medicine | 2002

Detection of cardiomyocyte apoptosis in forensic autopsy cases

Masato Nakatome; Ryoji Matoba; Yoshiaki Ogura; Zaw Tun; Mineo Iwasa; Yoshitaka Maeno; Hiroyoshi Koyama; Y. Nakamura; Hiroyuki Inoue

Abstract The purpose of the present study was to determine reliable parameters for the detection of apoptotic cells for use as a diagnostic marker during the early stage of acute myocardial infarction (AMI) in forensic autopsy cases. Myocardial tissues taken from forensic autopsy cases were examined by immunohistochemical and molecular-biological methods using the terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end-labelling (TUNEL) and the DNA laddering methods. In cases of AMI with a time period between 2 h from onset to death and 20 h post-mortem time, the nuclei of cardiomyocytes were stained positive with the TUNEL method and DNA fragmentation of myocardial cells was detected by agarose gel electrophoresis. Similar findings were obtained in cases of carbon monoxide (CO) intoxication. However, no apoptotic cells were found in other cases such as methamphetamine (MAP) intoxication, tetrodotoxin intoxication, alcohol intoxication, asphyxia, head injury, heart injury or myocarditis. These findings suggested that it would be possible to apply TUNEL-positive cells as a diagnostic marker during the early stages of AMI.


Forensic Science International | 2001

Examination of Y-STR mutations in sex chromosomal abnormality in forensic cases

Katsuya Honda; Zaw Tun; David Young; Toshiyuki Terao

The Y-STR typing was carried out on eight DNA samples (three from criminal cases) demonstrating Klinefelters syndrome. STR types in the X chromosome were randomly distributed. However, some Y-STR markers were distributed within the normal range but restricted to only one or two specific alleles, that is, some specific haplotypes were found in Klinefelters syndrome. In addition, a single nucleotide polymorphism in DYS390 (transversion of G to A at the 28th position downstream of tandem repeats) was detected in Klinefelter samples. This Y-STR polymorphism and restricted Y-STR alleles in Klinefelters syndrome is not known, but it might be related to the genesis of Klinefelters syndrome. We also found that extended standard haplotypes of these samples are extremely rare in the normal population, according to the Y-STR haplotype reference database (YHRD). The extended standard haplotype database in a Japanese population is also reported. In 100 unrelated Japanese, 89 haplotypes were observed, and the haplotype diversity was calculated to be 0.9866.


Journal of Forensic Sciences | 1996

Rapid and clear detection of ABO genotypes by simultaneous PCR-RFLP method.

Zaw Tun; Katsuya Honda; Masato Nakatome; Mohammed Nasimul Islam; H. Bai; Yoshiaki Ogura; Hisanaga Kuroki; Motohiko Yamazaki; Masaru Terada; Choei Wakasugi

We reported a new approach of ABO genotyping by a polymerase chain reaction and restriction fragment length polymorphism method. Instead of amplifying the loci containing the positions of nucleotides 258 and 700 of cDNA of the A transferase separately, we successfully amplified these 2 loci together in one reaction mixture using 2 sets of primers. The amplified DNA products were digested at the same time with restriction enzymes Kpn I and Alu I. The digested DNA products were then separated by electrophoresis on polyacrylamide gel. In addition, we evaluated the influence of various amplification parameters (concentration of template DNA, primers, Taq DNA polymerase, MgCl2, and number of cycles). In particular, high Mg2+ concentration (3.5 mM) made effective amplification of this locus without producing any unspecific band. By using that optimized condition for PCR, together with a simultaneous approach, our study proved to be time saving, more economic, and convenient in interpreting the results.


Journal of Forensic Sciences | 2001

DNA Testing of Klinefelter's Syndrome in a Criminal Case Using XY Chromosomal STR Multiplex-PCR

Katsuya Honda; Zaw Tun; Ryoji Matoba

We report genetic typing of Klinefelters syndrome applied to casework in forensic DNA testing. In this case, by using extracted DNA from body samples (muscle and bones), we could identify two distinct X alleles in two out of three X-STR loci (HPRTB and ARA), in addition to Y alleles (DYS390, DYS393). The extra X was found to have originated from father, and the victim turned out to have 47XXY Klinefelters syndrome. The victim was a 30-year-old male, born from relatively elderly parents as a second child. His father was a severe alcoholic and had been malnourished for more than 20 years at the moment of his birth. He exhibited slight mental retardation as a child, and belonged to a criminal group as an adult. The method presented here was useful to accurately diagnose sex chromosomal abnormality instead of conventional chromosomal analysis and Xg blood group typing. A subtype of this syndrome, 48 XXXY or mosaic, for example, could be identified if the intensity of the overlapped X bands were calculated.


Journal of Forensic Sciences | 1997

An electrocution death of an infant who had received an electric shock from an uncovered oval shaped lamp switch in his mouth while in a hospital

Motohiko Yamazaki; H. Bai; Zaw Tun; Yoshiaki Ogura; Choei Wakasugi

A male infant aged one year and nine months was found dead on a bed after admission to hospital with suspected pneumonia. The patient apparently put an uncovered oval shaped lamp switch (pendant switch) into his mouth and died of electric shock after contacting the exposed wires of the switch (100 V, 60 Hz alternating current). There were extensive first- to fourth-degree burns on the inner surface of the both lips. Because the histological findings were consistent with electric burns and the burns showed vital reactions, electric shock was judged to be the cause of death. The pendant switch is normally a very convenient piece of bedside equipment for inpatients. However, when the patient is an infant who naturally puts all the objects into the mouth, such a switch should be placed out of reach, and it should be certain that the cap is not loose.


Korean Journal of Biological Sciences | 2001

Analysis of the short tandem repeat Ioci for STRX 1, HPRTB, ARA, DYS390, DYS392 and DYS393 in Koreans

Hye Won Seol; Zaw Tun; Katsuya Honda; Shogo Msawa; Kyung Sook Park

Three STR loci (STRX 1[AGAT]n, HPRTB[AGAT]n and ARA[AGC]n) on X chromosome and three other STR loci (D YS390[CTG(A)T]n, DYS392[ATT]n and DYS393[GATA]n) on Y chromosome were analyzed in 154 unrelated healthy Korean subjects. Four loci (STRX1, HPRTB, DYS390 and DYS393) were amplified by quadruplex polymerase chain reaction (PCR) using fluorescent labeled primers (FLP). ARA and DYS392 were amplified separately using single PCR, similarly by using FLP. They were then run in an automated DNA sequencer and were analyzed with Genescan software. We found 7 alleles (308–332 bp) in STRX1, 7 alleles (275–299 bp) in HPRTB, 16 alleles (252–315 bp) in ARA, 6 alleles (203–223 bp) in DYS390, 7 alleles (245–263bp) in DYS392 and 5 alleles (116–132 bp) in DYS393. The *13 (34%), * 13 (51%), *23 (18%), *23 (50%), * 14 (39%) and * 13 (40%) alleles were observed to be the highest frequencies of STRX1, HPRTB, ARA, DYS390, DYS392 and DYS393, respectively. The detection of STR loci on sex chromosomes by quadruplex PCR allows simple determination of sex and identification of an individual.


Legal Medicine | 1999

An unusual autopsy case of a victim stabbed during sadomasochistic prostitution

Motohiko Yamazaki; Masaru Terada; Zaw Tun; Sachiko Takikita; Katsuya Honda; Ryoji Matoba; Tadahiko Shinohara

A 29-year-old male who lived alone was found dead with his back leaning against the wall of his room. He had been stabbed in his abdomen with a survival-type knife. The victims mouth was plugged with a pink-colored glove and was sealed with packing tape. His wrists were tightly tied behind his back with the same type of packing tape. The cause of death was diagnosed as hemorrhage from the right common iliac artery and vein. Due to the strange circumstances of the crime scene, the police initially considered the possibility of homicide by a group of assassins. Two months later, the police arrested a male suspect who was a member of a vice racketeer. The victim was characterized as a masochist and bisexual. He often hired the male suspect to perform sadomasochistic activities. On the day of the crime, the victim prepared a survival-type knife and packing tape himself to experience fear and pain more strongly. The victim hoped to use the knife to increase sexual excitement. In this case of sadomasochistic prostitution leading to death, the legal issues of homicide for money, malicious request of injury by the victim and accidental death were involved.


Archive | 1996

Detection of Sequence Variants in Hypervariable Segments of Mitochondrial DNA in the Asian Population

Katsuya Honda; Masato Nakatome; S. Harihara; Zaw Tun; Mohammed Nasimul Islam; H. Bai; Yoshiaki Ogura; Hisanaga Kuroki; Motohiko Yamazaki; Masaru Terada; Shogo Misawa; Choei Wakasugi

The analysis of highly polymorphic regions of mitochondrial DNA is one of the most commonly used methods for personal identification. The recent advances of fluorescent detection in automated DNA sequencing (Smith et al. 1986) has made it possible a rapid analysis of sequence variants without using isotopic labeling.


European Journal of Endocrinology | 2000

A novel mutation of the KAL1 gene in monozygotic twins with Kallmann syndrome

Takeshi Matsuo; Shingo Okamoto; Yukiko Izumi; Akiko Hosokawa; Takashi Takegawa; Hiroshi Fukui; Zaw Tun; Katsuya Honda; Ryoji Matoba; Ke-ita Tatsumi; Nobuyuki Amino

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