Public attitudes to the promotion of genomic crop studies in Japan: correlations between genomic literacy, trust, and favourable attitude.

This study aimed to assess public attitudes in Japan to the promotion of genomic selection in crop studies and to examine associated factors. We analysed data from a nationwide opinion survey. A total of 4,000 people were selected from the Japanese general population by a stratified two-phase sampling method, and 2,171 people participated by post; this survey asked about the pros and cons of crop-related genomic studies promotion, examined people's scientific literacy in genomics, and investigated factors thought to be related to genomic literacy and attitude.

Relationship between public attitudes toward genomic studies related to medicine and their level of genomic literacy in Japan.

The aim of this study was to assess public attitudes toward the promotion of genomic studies related to medicine and to examine the relationship between public attitudes and the level of genomic literacy by analyzing data from a nationwide opinion survey. The participants comprised 4,000 people (age, 20-69) selected from the Japanese general population by using the two-step stratified random sampling method. They were queried on the following topics in a mail survey: (1) pros and cons of the promotion of genomic studies related to medicine, (2) level of scientific literacy in genomics, (3) demographic and socioeconomic background, and (4) knowledge and attitudes toward science in general and genetic testing in particular.

IgG immunohistochemistry for the assessment of brain injuries in forensic autopsies.

Immunohistochemical staining of IgG in the sections of injured brain areas was performed in forensic autopsies. IgG immunoreactivity was present mainly in glial cells surrounding hemorrhagic areas, which may be a useful tool to detect and evaluate injured areas of the brain in forensic autopsies.

Immunohistochemical localization of heat shock protein 70 in the human medulla oblongata in forensic autopsies.

Heat shock protein 70 (hsp70) can be induced under various stresses in experimental animals. We investigated hsp70 immunoreactivity in the human medulla oblongata in forensic autopsies. Hsp70 immunoreactivity was observed in the cytoplasm of some neurons in the hypoglossal nucleus (XII), the dorsal motor nucleus of the vagal nerve (X), the lateral cuneate nucleus (Cun), and the inferior olive (Oli).

Immunohistochemical localization of P-selectin in the glomeruli from forensic autopsies.

The expression of the adhesion molecule P-selectin is known to be up-regulated in several vital organs including the kidney after trauma in experimental animals. We examined the expression of P-selectin in the kidney by immunohistochemistry in 41 forensic autopsies mainly from trauma cases. P-selectin immunoreactivity was present in the glomerular capillary endothelial tufts and cortical interstitial vascular endothelial cells.

Immunohistochemistry of atrial natriuretic peptide in brain infarction.

Atrial natriuretic peptide (ANP) was originally isolated from cardiac atria, and has potent natriuretic, diuretic, and vasorelaxant properties. It has been localized in neurons and astrocytes in the cerebral cortex and the white matter. We hypothesize that glial ANP may contribute to the regulation of cerebral blood flow in brain infarction.

An immunohistochemical study on cathepsin D in human hippocampus.

Cathepsin D is a lysosomal enzyme involved in neuronal degeneration. In this study, the immunohistochemistry of cathepsin D was studied in hippocampal CA1 neurons that are vulnerable to ischemia, and parahippocampal glial cells. CA1 neurons from the majority of cases showed cathepsin D immunoreactivity in the cytoplasm, whereas shrunk neurons were unstained in only one case.

Immunohistochemical localization of c-fos in the nuclei of the medulla oblongata in relation to asphyxia.

The immediately early gene product c-fos is known to be induced in neurons under noxious stimuli. Therefore, the immunohistochemistry of c-fos expression in human brains might offer information on the localization of stimulated neurons. In this study, the immunohistochemical localization of c-fos was studied in the neurons of the hypoglossal nucleus (XII), the dorsal motor nucleus of the vagal nerve (X), the nucleus solitarius (Sol), the accessory cuneate nucleus (Cun), the spinal trigeminal nucleus (V) and the inferior olive (Oli) of the human medulla oblongata from forensic autopsy cases.

[The structural polymorphism in D8S580-STR region].

The STR structure of the D8S580 locus was analyzed by the base sequencing technique. Alleles were collected separately by urea denaturing polyacrylamide gel electrophoresis of the PCR amplification product, followed by cutting of the target DNA band from the gel, and reamplification. As a result, this locus was shown to have a complex STR structure consisting of four types of repeat units, i.

Immunohistochemistry of neuron-specific enolase in neurons of the medulla oblongata from human autopsies.

Neuron-specific enolase (NSE) is a glycolytic enzyme specifically expressed in neurons. NSE has been used as a marker for neuronal damage in brain injury. We studied the immunohistochemical localization of this enzyme in the medulla oblongata obtained from human forensic autopsy specimens.

Immunohistochemical study of neuron-specific enolase in human brains from forensic autopsies.

Immunohistochemistry using anti-human neuron-specific enolase (NSE) mouse monoclonal antibody was performed in human brains from autopsy cases, which enabled us to assess the neuronal damage besides hematoxylin and eosin or Klüver-Barrera stain. Neurons in cerebral neocortex which showed necrotic changes such as prominent cytoplasmic vacuolization or cellular shrinkage with nuclear pyknosis showed a tendency to be less stained by anti-NSE antibody. Anti-NSE immunostaining was statistically significantly less in the neocortex from CO intoxication than from other causes of death, although morphological necrotic changes were less observed in CO intoxication.

[Structural polymorphism of ACTBP2 STR region].

We investigated structural polymorphisms of the ACTBP2-STR region by sequence analysis instead of using PCR-amplified fragment length polymorphisms. A gel spot extraction reamplification method using 2% agarose gel was effective for preparing the individual alleles from zygotes with a length difference of more than 16bp. In a survey of polymorphisms in 116 unrelated Japanese subjects, alleles from 73 zygotes were prepared by this method.

[HLA-DQB2 genotypes analyzed by PCR/SSCP method].

When the PCR products amplified by the primers prepared at the 11th HLA Workshop (DQBAMP-A, DQBAMP-B) were analyzed directly by the SSCP method, one or two pairs of characteristic bands were detected other than those attributed to DQB1, and a total of three kind of paired bands were detected. To confirm that these bands were allelic genes of DQB2, the corresponding bands were isolated by cloning, and their base sequences were determined. The base sequence of one of them was in agreement with that of DX beta, which has already been described, and the characteristic 3-base defect was noted by comparison with the base sequence of DQB1.

[Sequence polymorphism of mitochondrial DNA and its forensic application].

We have investigated polymorphism of mitochondrial (mt) DNAs in the Japanese population. In order to compare 288-bp sequences (nucleotide positions 16,111 to 16,398) in the noncoding region, a 452-bp segment elongated by 82 bp at both sides of the target was amplified by PCR and analyzed directly by Taq cycle sequencing with FITC-labelled primers. A survey of 100 Japanese individuals revealed the existence of 66 types of mtDNAs.

Polymerase chain reaction (PCR)/single-strand conformation polymorphism (SSCP) analysis of the human HLA-DQB regions.

Genetic diagnosis of 13 alleles of HLA-DQB1 (0501, 0502, 5031, 5032, 0601, 0602, 0604, 0201, 0301, 0302, 3032, 0401 and 0402) from 65 human DNA samples was achieved by applying single-strand conformation polymorphism (SSCP) analysis to DNA fragments amplified by the polymerase chain reaction (PCR) using a convenient primer set for DQB1 (recommendation of the International Histocompatibility Workshop, 1991). Differences between strand images (narrow/distinct or broad/diffuse) from the individual alleles and their electrophoretic mobilities are regarded as criteria for confirming the genetic diagnosis of DQB1 alleles. This primer set amplifies not only DNA fragments belonging to DQB1, but also to DQB2, and classification of 3 phenotypes (1.

Detection of carboxyhemoglobin using Fourier transform infrared microspectroscopy.

We investigated the absorption of carboxyhemoglobin (COHb) using a Fourier transform infrared (FTIR) microscopy system. Weak absorption at 1969 cm-1 due to Fe<--CO and strong absorption due to N-H stretching vibration, > C = O stretching vibration and -NH bending vibration (1544 cm-1) were observed. The ratio of absorbance at 1969 cm-1/1544 cm-1 can be used as an index of CO saturation.

[Water-soluble eumelanin as a PCR-inhibitor and a simple method for its removal].

It has been confirmed that water-soluble eumelanins often extracted together with DNAs from natural black hairs act as an inhibitor of Taq DNA polymerase in the polymerase chain reaction (PCR). In the present investigation, an attempt to amplify the non-coding 333-bp region of mitochondrial DNA (mt333DNA) produced the following results: 1) Water-soluble preparations made from chemically synthesized melanin (Sigma products), as well as natural black eumelanins, inhibited the PCR amplification of mt333DNA at concentrations of more than 2 micrograms/ml. 2) Quantitative measurement of Taq DNA polymerase-catalyzed DNA synthesis in terms of the amount of [alpha-32P] dCMP incorporated into activated calf thymus DNA showed that both of the water-soluble melanins had the same inhibition activity as represented by the sigmoidal curve derived from a quadratic equation of melanin concentration.

[Presence of a PCR-inhibitor in hairs].

A polymerase chain reaction (PCR) system was used to amplify the noncoding 333-bp region of mitochondrial DNA (mt333DNA) contained in DNA extracts from single human hairs, and the following results were obtained: 1) Using natural black hairs, mt333 DNA was always amplified from a 5-cm length of hair shaft sampled within a region 11 cm from the hair root, but it was not always amplified from a 5-cm region adjacent to this 11-cm region, and was not amplified in almost all cases when a 5-cm length of hair shaft was sampled from a region more than 16 cm distant from the hair root. DNA preparations not responding to PCR were colored dark brown. 2) Using natural white hairs, mt333DNA was amplified from almost all specimens even up to a length of 31 cm.

[Three cases of acute methamphetamine intoxication--analysis of optically active methamphetamine].

Three typical cases of acute methamphetamine intoxication are reported. The concentrations of methamphetamine in the blood were 27.2, 6.

The effect of cyanide on isolated perfused rat heart.

Using Langendorff isolated perfused rat hearts, we demonstrated direct effects of cyanide on the heart. A heart from a sacrificed male rat was placed on a Langendorff apparatus and perfused with Tyrode solution containing 2.0 mM NaCN (CN-TS), following with normal Tyrode solution (TS) at 37 degrees C.

[Individual identification from extremely contaminated forensic specimens by sequencing cloned mitochondrial D-loop genes].

The PCR-amplified fragments of the human mitochondrial DNA were cloned, and independent clones were sequenced to identify individuals from trace amount of a composite forensic specimen originated from plural number of the individuals. The amplification of the mitochondrial DNA is suitable for forensic analysis where the specimens are highly degraded in most cases for its extremely high copy number and polymorphism as compared with the chromosomal DNA. And cloning procedure simplifies to correspond results to each individuals.

[Quantitation of hemagglutinins using new microtitration plate (R-plate) and ELISA reader].

A new type of microtitration plate (reverse-shaped bottom plate, R-plate) was used in quantitative hemagglutination test. The bottom of the well of this plate was shaped convex and the hemagglutination patterns were measured as photometric value by the ELISA reader. Agglutinated erythrocytes adhered to the convex bottom of the well and were detected as large absorbance.

Estimation of optically active methamphetamine and amphetamine using alpha-methoxy-alpha-trifluoromethylphenylacetyl chloride.

A simple analytical method for determination of optically active amphetamine and methamphetamine using (+)-alpha-methoxy-alpha-trifluoromethylphenylacetyl chloride (MTPA-Cl) was developed. The method was found to be useful for determining the absolute configuration and optical purity. The results obtained using the capillary GC method agreed well with those of the NMR method.

Reduction of nitroolefin using microorganisms.

Microbial reduction of 1-phenyl-2-nitro-1-propene (3) was carried out using 57 strains of yeast, 40 strains of aerobic and facultatively anaerobic bacteria and 40 strains of strictly anaerobic bacteria. Nine strains of yeast (Candida tropicalis, etc.,) had the ability to reduce (3) to 1-phenyl-2-nitropropane (1) (94.

[Homicidal poisoning by aconite: report of a case from the viewpoint of clinical forensic medicine].

A case of homicidal poisoning by aconite is reported from the viewpoint of clinical forensic medicine and analytical chemistry. Jesaconitine was detected in the vomitus, stomach contents, plasma and urine at concentrations of 32.2, 5.