The Effect of Hypothermia on the Gene Expression of Inducible NOS and NF-kappaB in the Lung in Hemorrhagic Shock in Rats

PURPOSE: In previous studies, therapeutic hypothermia (HT) in hemorrhagic shock (HS) had beneficial effects on the hemodynamic and metabolic parameters, and on the survival. The mechanism is uncertain. We hypothesized that the suppression of inducible nitric oxide synthase (iNOS) and NF-kappaB could be associated with the beneficial effects of therapeutic HT in HS. METHODS: Sixteen male Sprague-Dawley rats were randomized to normothermia (36~37degrees C, NT group) or moderate hypothermia (27~30degrees C, HT group). They underwent volumecontrolled (2 ml/100 g weight) HS (90 minutes) and partial resuscitation with shed blood (1 ml/100 g). RESULTS: In the HS and post-resuscitation phase, the mean arterial pressures were higher in the HT group than in the NT group and PaO2 was higher in the HT group than in the NT group. The lactate level was lower in the HT group than in the NT group (1.1+/-1.1 nmol/L vs 6.4+/-5.0 nmol/L, p=0.021). Serum concentrations of IL-1 beta and IL-6 and activated p65 NF-kappaB levels in the lung tissue were higher in NT group than in the HT group (p<0.05). Lung malondialdehyde contents and the expression of iNOS mRNA were significantly decreased in the HT group compared to the NT group (63.8+/-6.2 nmol/g vs 44.6+/-4.5 nmol/g, p<0.001; 1313.0+/-924.4 vs 9088.4+/-3984.0 arbitrary units, p<0.001, respectively). CONCLUSION: These data suggest that in HS, therapeutic HT inhibits lipid peroxidation, activation of NF-kappaB, and gene expression of iNOS in the lung. These factors might be the mechanism of the beneficial effects of HT in HS.

Reliability and Validity of the Korean Version of the Psychotic Symptom Rating Scale / 신경정신의학

OBJECTIVES: Psychotic Symptom Rating Scale (PSYRATS) is an assessment tool to measure the severity of different dimensions of auditory hallucinations and delusions. The reliability and validity of the Korean version of PSYRATS (K-PSYRATS) were examined in Korean patients with major psychosis. METHODS: The inter-rater reliability of the K-PSYRATS was determined from the videotaped interviews of the five schizophrenic patients. To measure validity and internal consistency reliability, the 109 patients with auditory hallucinations or delusions were assessed using the K-PSYRATS, Positive and Negative Syndrome Scale (PANSS) and Clinical Global Impression-Severity (CGI-S) scale. RESULTS: K-PSYRATS was found to have excellent inter-rater reliability (intra-class correlation coefficient of auditory hallucination= 0.81, p<.001, intra-class correlation coefficient of delusion=0.97, p<.001) and internal consistency reliability (Cronbach's alpha of auditory hallucination=0.77, Cronbach's alpha of delusion=0.76). Significant correlation was found between K-PSYRATS and positive syndrome subscale of PANSS and CGI. CONCLUSION: K-PSYRATS is a useful assessment instrument for psychotic symptoms in Korea.

State of Individual Identification Using Genetic Polymorphism in Korea: Step to Criminal DNA Bank / 대한법의학회지

There have been several attempts for criminal DNA profiling in Korea, but ended without any fruitful results. Recently new movement for the profiling has just been started. Many have to be solved in advance, which can be divided largely into two, technical aspects and legal system which could support the application. To build up profiling for any purpose, work for the standardization must be driven, because many are involved in the work. Authors have checked the status of the DNA typing in Korea and have reviewed its quality by statistical approach. All the popular forensic journals, Korean or international covering from 1997 to 2004, were reviewed to check whether it contains articles on Korean. All the articles containing genetic data on Korean were selected and their results were compared statistically. Total 67 articles were chosen. Articles were on 116 loci, among which 54 were autosomal, 32 were Y chromosomal and the remaining were X chromosomal. Among 32 Y chromosomal loci, 11 were bi-allelic. For the 50 loci two or more lab reported separately. For these loci the results from different lab were compared. There was no statistically significant difference for most of the loci, but some of the loci, especially on Y chromosomal STR loci, showed some differences. Data were showed with some review.

Molecular Characteristics of B Subgroups in Koreans / 대한진단검사의학회지

BACKGROUND: An accurate ABO blood type is essential for transfusion therapy. Genetic structures of ABO blood group and subgroup have been investigated and so far about 100 ABO alleles have been reported worldwide. This study was performed to investigate the molecular characteristics of B subgroups in the Korean population. METHODS: A total of 19 samples of B subgroups were collected from patients (n=11) and from blood donors (n=8) of Korean Red Cross blood centers; these samples had been typed serologically for the ABO blood group. Allele-specific polymerase chain reaction (PCR), direct sequencing of exon 6 and 7, and allele separation were performed for ABO gene analysis. RESULTS: The ABO PCR-RFLP genotyping results of 18 samples among the provisional 19 B subgroups were identical regardless of their phenotypes. Two new B alleles showing 255C>T base change and 547G>A base change were observed in B3 and A1B3 subgroups. CONCLUSIONS: Serologically unidentified B subgroups were unequivocally identified through molecular analyses of the ABO gene. And new ABO alleles observed only in the Korean B subgroups were recognized.

Heteroplasmy Pattern Within D-loop Region of Mitochondrial DNA in Koreans / 대한법의학회지

Mitochondrial DNA (mtDNA) sequencing has been validated as a useful tool in forensics. However, there are several aspects that need to be considered in order for it to be used as firm evidence in a courtroom. Heteroplasmy is one of those. Heteroplasmy means the status that different mtDNA populations exist in a single individual, tissue, cell or even mitochondrion. As the underlying phenomenon of heteroplasmy and polymorphism is similar, care must be taken in analyzing the data from the samples with heteroplasmy. Heteroplasmy can reinforce the use of mtDNA however if we know a lot about it. For this much has to be revealed. In this we screened 494 unrelated Korean using DGGE for the D-loop to determine how frequent heteroplasmy is, and the heteroplasmy pattern was confirmed through sequencing. This paper discussed the results of a comparison with other reports along with several aspects of heteroplasmy.

Bi-allelic loci on Y chromosome in Korean / 대한법의학회지

Recently studies about bi-allelic markers such as SNP, which is commonly found in about every 1.2 kb, seem to be increasing. Compared to STR marker, much have to be improved if it is to be used for individual identification. Nevertheless many researchers have interests in SNP and it 's scope is unpre-dictable. SNP may be promising as an auxiliary tool in individual identification, especially in Y chromo-somal study, in which the usefulness of conventional STR markers are restricted as the concept of haplo-type is applied. We report allelic distribution pattern in Korean for several previously known bi-allelic markers, that are SY81, M9, SRY1532, SRY2627, YAP. In three loci that are SY81, SRY1532, SRY2627, no polymorphism was noted. In M9, YAP loci, bi-allel-ic polymorphism was noted. In M9, 79.3% was G-type, and C-type was 20.7%. The YAP insertion was positive only in 6%. Remaining 94% was YAP(-). These pattern was compared with that of other popu-lation, and racial difference was evident. Several key points about SNP were discussed.

Molecular Characteristics of A Subgroups in Koreans / 대한수혈학회지

BACKGROUND: An exact ABO blood type is essential for transfusion therapy. Genetic structures of ABO blood group and subgroup have been investigated and about 100 ABO alleles have been reported worldwide. This study was performed to investigate the molecular characteristics of A subgroups in the Korean population. METHODS: Nine A and five AB subgroups were collected from patients and from blood donors of Korean Red Cross blood centers after serological ABO blood group typing. On these samples, allele-specific polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP), direct sequencing of exon 6 and 7, and allele separation were performed for ABO gene analysis. RESULTS: The ABO PCR-RFLP genotyping results of 13 samples among the provisional 14 A subgroups were identical with their phenotypes. ABO*A205 allele was observed in an Aint subgroup. Two new A alleles that showed 784G>A base change and 990C>T of intron 6. And a polymorphism of 532C>T in A(pro) intron 5 were also discovered. Conclusion: Through the molecular analysis of this study, serologically unidentified A subgroups were obviously identified. And the new alleles only observed in the Korean A subgroups were recognized.

Development of Multi-tissue Array Block for Diagnosis of Early Myocardial Infarction / 대한법의학회지

Tissue array contains hundreds or thousands tissues from different cases, and has been known useful for rapid analysis of markers in a large numbered work. This has mainly been used in investigative purpose, and few has been reported for diagnostic trial. Diagnosis of early myocardial infarction is important in medicolegal field. Immunohistochemical staining using various antibodies has become a popular tool for this purpose, but standardization and reproducibility have been problems. Use of tissue array is promising in this points. We have made two cardiac tissue arrays using 87 different cases and have verified the feasibility of tissue array in forensic field. Experiences and several issues concerning tissue array are discussed.

Analysis of Five Coding Region Polymorphism in Mitochondrial DNA / 대한법의학회지

For evaluation of the five coding region (CR) polymorphism in mitochondrial DNA (mtDNA); we had performed PCR and direct sequencing in 599 unrelated Korean who showed the identical DNA type in D-loop mitochondrial DNA analysis for total 2,810 bp fragment. Following the sequence analysis, all the sequences of five regions were compared respectively to Anderson standard sequence to investigate the nucleotide variations. The result showed, a total 4,565 nucleotide variations were observed at 190 positions in five CR as 3,931 (86.11%) substitutions, 32 (0.7%) insertions, and 602 (13.19%) deletions and the allele diversities (h) were higher than 0.9992 when adding each CR or combined CR to D-loop analysis in mtDNA. In conclusion, we could confirm the five CR are useful for forensic testing through the nucleotide variation and hapolotypes polymorphism.

Polymorphism and Haplotypes of 10 Y-STR Loci in Koreans / 대한법의학회지

Ten Y chromosomal STRs (DYS434, DYS435, DYS437, DYS438, DYS439, DYS441, DYS442, DYS443, DYS444, DYS445) have been typed in 1159 Korean males for the purpose of application to male identification, population genetics and evolution study. Multiplex PCR method was used in amplifying simultaneously these loci. In each locus, 3-8 allele were observed, and no 'interallele' was observed. The gene diversity varied from 0.3131 at DYS435 to 0.7179 at DYS441. Among 610 father-son pairs, mutation was observed 2 cases in DYS434, 1 case in DYS435, 3 cases in DYS437, 1 case in DYS439, 2 cases in DYS441, 1 case in DYS442 and 4 cases in DYS444. No two or more simultaneous mutations were identified in one father-son family pair. In 1159 Korean males, 641 different haplotypes were observed, and 488 haplotypes were not shared by others. The haplotype diversity was 0.9911, Probability of Identity was 0.0098. When using the ten loci together with DYS19, DYS385, DYS388, DYS389, DYS390, DYS391, DYS392, DYS393 which were already used in the Department of Forensic Medicine, College of Medicine, Seoul National University, the haplotype diversity was 0.9997, Probability of Identity was 0.0012.

Death Associate with Sweet's Syndrome / 대한법의학회지

Sweet's syndrome was originally described as an "acute febrile neutrophilic dermatosis". Sweet's syndrome presents in three clinically settings: (1) classical or idiopathic Sweet's syndrome, (2) malignancyassociated or paraneoplastic Sweet's syndrome, (3) drug-induced Sweet's syndrome. Sweet's syndrome is known to respond well to systemic corticosteroids, and mortal case associated without any other predisposing factors is hardly found. Twenty-nine year old male was transferred from local clinic with the chief complaint of recurrent thrombophlebitis which occurred 10 days ago after IV treatment of antibiotics for burn. The symptom did not get well with conventional treatment for the thrombophlebitis such as antibiotics or drainage. After several days of pyrexia, the patient died suddenly. Grossly abnormal findings including pulmonary thrombus which could explain sudden death were not found. Disseminated intravascular microthrombi was the most peculiar finding. It was prominent in small sized vein, and lung was the most severely affected. Heart and liver were also involved. Large thrombi within deep vein of the legs were also noted. Minor collections of inflammatory cells, even apart from the main lesion, were noted in skin adnexa or small vessles within aorta wall. No tumorous area was found. The disseminated intravascular coagulation, which was associated with vasculitis was thought to be the direct cause of death. Several points about the Sweet's syndrome including the unusualness of this case were discussed.

Analysis of Polymorphism of Penta E and Penta D in Korean / 대한법의학회지

A duplex PCR reactions were designed for two pentameric (Penta D, Penta E) short tandem repeats (STR) loci to study the allele frequencies and the applicability of genetic variation in these loci to forensic case works. For 323 unrelated Koreans DNA was extracted using Qiagen DNA blood kit. The fluorescently labeled amplified products were analyzed by 8M urea, 5% long range gel electrophoresis in ABI-377 sequencer and followed data analysis by Genescan Analysis and Genotyper DNA fragment analysis software. The PD, PIC and PE were above 0.92, 0.76 and 0.66, respectively. No deviation from Hardy-Weinberg equilibrium was observed of Penta D locus (p=0.35, observed number>1).

DGGE assay for heteroplasmy in mitochondrial DNA / 대한법의학회지

Heteroplasmy in the hypervariable region 1 of the mtDNA was screened using DGGE. Blood samples from 150 unrelated Koreans were used. Appropriate condition was set up using mixed samples with previously known HV1 sequences. This method is capable of detecting heteroplasmic proportion less than 5 %. Multiple bands pattern meaning heteroplasmy was observed in 41 individuals (27.3%). The majority of heteroplasmic variants could not be detected by direct sequencing of PCR products. For the distribution pattern of heteroplasmy in different tissues within one individual, 14 autopsy cases were screened. Out of these, two were heteroplasmic, but there was no difference between various tissues in the heteroplasmic DGGE band pattern.

Comparison of STR Typing Results from Several Centers for the Same Samples: Steps to Standardization for STR Typing / 대한법의학회지

This paper described a collaborative exercise intended to see what kinds of short tandem repeat (STR) loci are used in different DNA typing laboratories in Korea and to compare their results for the demonstration whether uniformity of DNA profiling results from different laboratory could be achieved in Korea. Laboratories were asked to test five tissue DNAs using methods routinely used in each laboratory and to report the results to the coordinating laboratory. The exercise demonstrated that each laboratory was using different STR loci for the typing with different STR numbers, 2 VNTRs, 36 STRs and amelogenin in total, and the direct comparison of the results from all the laboratory for the 18 loci could not be done as only one laboratory submitted typing results. Among 21 loci for which several laboratories submitted typing results, results for 14 loci were the same and results for the other 7 loci were different depending on the participating laboratory. D1S80, F13A01, D16S539, D21S11, D18S51, D3S1744 were the loci with different typing results. Even in the cases where commercial kits were used, the results were not the same depending on the machines used, that is the capillary electrophoresis or the gel based electrophoresis. The reason for the different results, points about the standardization of the methods and the profiling data were described.

Comparison of STR Typing Results from Several Centers for the Same Samples: Steps to Standardization for STR Typing / 대한법의학회지

This paper described a collaborative exercise intended to see what kinds of short tandem repeat (STR) loci are used in different DNA typing laboratories in Korea and to compare their results for the demonstration whether uniformity of DNA profiling results from different laboratory could be achieved in Korea. Laboratories were asked to test five tissue DNAs using methods routinely used in each laboratory and to report the results to the coordinating laboratory. The exercise demonstrated that each laboratory was using different STR loci for the typing with different STR numbers, 2 VNTRs, 36 STRs and amelogenin in total, and the direct comparison of the results from all the laboratory for the 18 loci could not be done as only one laboratory submitted typing results. Among 21 loci for which several laboratories submitted typing results, results for 14 loci were the same and results for the other 7 loci were different depending on the participating laboratory. D1S80, F13A01, D16S539, D21S11, D18S51, D3S1744 were the loci with different typing results. Even in the cases where commercial kits were used, the results were not the same depending on the machines used, that is the capillary electrophoresis or the gel based electrophoresis. The reason for the different results, points about the standardization of the methods and the profiling data were described.

Quadruplex PCR for 8 Autosomal STR Loci in Korean / 대한법의학회지

Two quadruplex PCR reactions were designed for 7 tetrameric (D3S2406, D4S2368, D5S818, D7S821, D9S925, D13S317, D19S253) and one trimeric (D6S1043) short tandem repeats loci to study the allele frequency and the applicability of genetic variation in these loci in forensic case works. For 310 unrelated Koreans DNA was isolated from peripheral blood using phenol/chloroform method. Quadruplex I was consisted of D4S2368, D6S1043, D7S821, D9S925 and quadruplex II D3S2406, D5S818, D13S317, D19S253. The amplified products were analyzed by 5%polyacrylamide gel electrophoresis followed by silver staining. The heterozygosity in each loci ranged 92.91-66.13%, and PD(Power of Discrimination) was above 0.85 in each loci. Every loci except D6S1043 followed hardy-Weinberg expectation. The cumulative PI was low as 1.65x10-10. Two mutations were noted, one in D19S253 and the other in D9S925 among 234 gametes. With these results above eight STR loci studied here proved to be highly polymorphic enough to be used in forensic field. This study provides valuable population data in these loci for Korean.

Quadruplex PCR for 8 Autosomal STR Loci in Korean / 대한법의학회지

Two quadruplex PCR reactions were designed for 7 tetrameric (D3S2406, D4S2368, D5S818, D7S821, D9S925, D13S317, D19S253) and one trimeric (D6S1043) short tandem repeats loci to study the allele frequency and the applicability of genetic variation in these loci in forensic case works. For 310 unrelated Koreans DNA was isolated from peripheral blood using phenol/chloroform method. Quadruplex I was consisted of D4S2368, D6S1043, D7S821, D9S925 and quadruplex II D3S2406, D5S818, D13S317, D19S253. The amplified products were analyzed by 5%polyacrylamide gel electrophoresis followed by silver staining. The heterozygosity in each loci ranged 92.91-66.13%, and PD(Power of Discrimination) was above 0.85 in each loci. Every loci except D6S1043 followed hardy-Weinberg expectation. The cumulative PI was low as 1.65x10-10. Two mutations were noted, one in D19S253 and the other in D9S925 among 234 gametes. With these results above eight STR loci studied here proved to be highly polymorphic enough to be used in forensic field. This study provides valuable population data in these loci for Korean.

Y Chromosome Haplotypes in Koreans / 대한법의학회지

In this study the population data at seven STR loci on the Y chromosome, DYS19, DYS388, DYS389, DYS390, DYS391, DYS392 and DYS393 are described for 1054 Koreans. In each locus, 6-22 alleles were noted, and allelic distribution patterns were found to be different from those of other populations. The PD was 0.28-0.886 and no interallele was noted. In 388 father-son pairs, 9 cases of mutation, one in DYS19 locus, one in DYS388, two in DYS389, three in DYS391, one in DYS392 and one in DYS393 locus were noted. In total 563 different haplotypes were noted. 630 cases shared the same haplotype with someone among 1054 object studied. Even in case which showed different haplotypes, many cases showed differences only in one locus and genotypes in the remaining seven loci were the same. The discrimination between mutation and different haplotypes seems to be problematic in these situations. Experiences for the large scale haplotype data base in Koreans were described.

Y Chromosome Haplotypes in Koreans / 대한법의학회지

In this study the population data at seven STR loci on the Y chromosome, DYS19, DYS388, DYS389, DYS390, DYS391, DYS392 and DYS393 are described for 1054 Koreans. In each locus, 6-22 alleles were noted, and allelic distribution patterns were found to be different from those of other populations. The PD was 0.28-0.886 and no interallele was noted. In 388 father-son pairs, 9 cases of mutation, one in DYS19 locus, one in DYS388, two in DYS389, three in DYS391, one in DYS392 and one in DYS393 locus were noted. In total 563 different haplotypes were noted. 630 cases shared the same haplotype with someone among 1054 object studied. Even in case which showed different haplotypes, many cases showed differences only in one locus and genotypes in the remaining seven loci were the same. The discrimination between mutation and different haplotypes seems to be problematic in these situations. Experiences for the large scale haplotype data base in Koreans were described.

Species Identification Using Cytochrome B Gene / 대한법의학회지

The feasibility of species identification using sequence analysis of the cytochrome B (Cyt B) gene in mitochondrial DNA was investigated. DNA was extracted from nine different animals that could be easily met in our surroundings and Cyt B gene was amplified. Direct sequencing results for the amplified PCR products were compared with each other. Human was also included. Nucleotide sequence of the Cyt B gene for each animals was also compared with the previously known ones registered in nucleotide databases, Genebank. The inter-species sequence variation was high as the percent similarity of each sequences ranged 64.6-83.5%. Compared to this the percent similarity of sequences obtained here were high when compared to the sequences of the same species registered in the database showing relatively low intra-species variation. This data shows that the nucleotide sequences of Cyt B gene in a certain biological materials can be identified at species level. The applicability of this method to the forensic field is also demonstrated by performing a casework; determination of the origin for the placentae which were commercial available as "invogorant". Points about the use of Cyt B gene in forensic field was also reviewed.