Long-term ethanol consumption in ICR mice causes mammary tumor in females and liver fibrosis in males.

BACKGROUND: Although epidemiological studies indicate that ethanol consumption and the risk of breast cancer are positively associated in women, experimental animal models have not yet been developed that provide evidence to support this relationship. To clarify alcohol-related liver injury, it is important to reproduce, in laboratory small animals, the liver fibrosis observed in human alcoholics. However, in mice the induction of fibrosis has failed. The present study describes the first experimental models to produce mammary tumors in female ICR mice and liver fibrosis in male ICR mice treated long-term with ethanol. METHODS: The study consisted of two parts. To induce mammary tumors, female ICR mice were given 10% to 15% ethanol solution as the sole drinking fluid for 25 months, with solid diet supplied ad libitum. To induce liver fibrosis, male ICR mice were given 10% to 15% ethanol solution as the sole drinking fluid for 10 to 15 months. Control female and male mice were given tap water. RESULTS: In 9 (45%) of 20 ethanol-treated female mice, mammary tumors occurred at 8 to 24 months after ethanol intake began, whereas spontaneous mammary tumor was not found in the 20 control female mice. The tumors were composed histopathologically of either papillary adenocarcinoma or medullary adenocarcinoma of glandular epithelial origin. In the ethanol-treated male mice, early hepatic fibrosis at the centrilobular and pericellular areas and central-central bridging were observed at the 10th month, and marked fibrosis at the centrilobular, pericellular, and periportal areas and bridging between the neighboring vascular tissues were observed at the 15th month, which suggested that the initial fibrosis arose from the centrilobular area. No abnormalities other than mild fatty infiltration were found in livers of the control male mice. CONCLUSIONS: These murine models may be useful to study the role of ethanol in mammary tumorigenesis and the pathogenetic mechanisms of ethanol liver injury.

Intralobular distribution of class I alcohol dehydrogenase and aldehyde dehydrogenase 2 activities in the hamster liver.

The hepatic lobular localization of class I alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) 2 activities was examined histochemically using livers of hamsters with high ethanol preferences. The activity of class I ADH detected by the nitro blue tetrazolium method using 5 mM ethanol as a substrate was extremely high and was almost homogeneously distributed throughout the lobule. The ALDH 2 activity (substrate, 8 microM acetaldehyde) was localized to the centrilobular zone, whereas low Km ALDH (ALDH 1 + ALDH 2) activity (substrate, 50 microM acetaldehyde) showed a gradient distribution in the lobule with high centrilobular to moderate periportal activity, suggesting that the ALDH 1 activity was distributed throughout the lobule.

Rapid detection of dihydrocodeine by thermospray mass spectrometry.

Rapid assay of dihydrocodeine (DHC) by thermospray mass spectrometry is explored. Liquid-liquid extractions of blood, urine and gastric contents were injected into a thermospray mass spectrometer, to which there was no column connected, and DHC was assayed by the flow injection method. The mass spectra of DHC under thermospray ionization and filament-on ionization modes consist of the MH+ ion of mlz 302 alone, which was clearly detected in the samples. Although DHC should be quantitated by gas chromatography-mass spectrometry, this method is applicable for rapid identification of DHC in biological materials.

PCR-based genotyping of MNSs blood group: subtyping of M allele to MG and MT.

PCR-based genotyping of MNSs blood group system was investigated in combination with restriction fragment length polymorphism (RFLP), single-stand conformation polymorphism (SSCP) and allele-specific PCR amplification (ASPA) techniques. M and N alleles are based on three nucleotide substitutions in exon 2 and one base change (G or T) in an intron of glycophorin A locus. The latter single base change was also found among M alleles analyzed in this study, so that M allele appeared to be subdivided into MG and MT. All three alleles, MG, MT and N were identified clearly by RFLP or SSCP analysis following a single amplification. S and s alleles are based on one nucleotide substitution in exon 3 of glycophorin B gene. Genotyping of Ss blood group system was also explored by PCR-SSCP or ASPA analysis, and problems in the methods were discussed.

Analysis and classification of nasopharyngeal electroencephalogram in "brain death" patients.

Nasopharyngeally-derived electroencephalogram (EEG) was recorded and digitized in 12 "brain death" subjects with flat-line scalp EEG and loss of auditory brain stem response. The nasopharyngeal EEGs of these cases were classified into three types: Type Ia with complete flat-line, Type Ib with almost but incomplete flat-line EEG, and Type II with low-amplitude slow fluctuations. Digitization of the nasopharyngeal EEG showed that equivalent electric potentials in low frequency bands (delta and/or theta 1) remained within the values of healthy volunteers in Types Ib and II. These results suggested that the tissue in or around the brain stem still functioned in Type 1b and II "brain death" patients. The origin of nasopharyngeal EEG was also discussed in this paper.

ABO genotyping following a single PCR amplification.

Using primers designed by Lee and Chang, 200 base-pair (bp) fragment of ABO locus was amplified by PCR, which spans the site of the single nucleotide deletion associated with O allele. O allele could be identified by Kpn I digestion of the PCR product as reported. A and B alleles were also distinguishable by Mae II digestion of the product. Thus restriction digestion by Kpn I and Mae II could genotype ABO blood group following the single amplification. The nucleotide substitution in the 200-bp product between A and B alleles was also found in O allele, resulting in 2 different suballeles OA and OG. The single-strand conformational polymorphism of the PCR product was also investigated for ABO genotyping following the single amplification.

Analysis of organic compounds in a case of suicide by ignition with lacquer thinner.

An autopsy case of suicide by ignition using lacquer thinner is presented. A wholly-charred body of a 54-year-old man and a can of lacquer thinner were found at the burnt driver's seat of a truck. Organic solvents in blood, urine, lung tissue, trachea and gastric contents and of the remaining clothes have been analyzed by gas chromatography/mass spectrometry (GC/MS). High levels of toluene, ethyl acetate and butyl acetate were detected in his clothes. The concentrations of toluene in the left and right heart blood, urine, gastric contents, squeeze sample and a block of lung tissue were 0.309, 0.226, 0.018, 0.051, and 0.268 mg/ml, and 0.340 mg/g, respectively. The ethanol levels in these samples were 1.89, 1.71, 1.58, 13.88 and 1.39 mg/ml, and 1.49 mg/g, respectively, and the profile suggested that the source of the ethanol was mainly drinking. The carboxyhemoglobin concentration in the left and right heart blood was 43.3 and 36.1%, respectively. The GC/MS data on organic solvents are consistent with the idea that he used the lacquer thinner contained in the can found in his truck for ignition. The high levels of toluene in his blood suggest that not only burns but also toluene poisoning contributed to his death.

Detection of green algae (Chlorophyceae) for the diagnosis of drowning.

The plankton test (generally, diatom test) is one of the methods available to diagnose the cause of death of submerged bodies. The solubilization method using tissue solubilizer Soluene-350 was used in this study to detect not only diatoms but also green algae, based on the fact that the solubilizer does not digest the cell walls of green algae which are made from cellulose. Detection of green algae from organs of submerged cadavers is very informative to determine drowning in fresh water, and also in cases where only few diatoms are detected in the organs.

[Clinical study on judgement of brain death by the nasopharyngeal lead with the aid of an automatic EEG analysis system].

We investigated the relationship between scalp electroencephalogram (EEG) and nasopharyngeal EEG using Automatic EEG Analysis System in 10 patients with the age between 17 and 66 who were clinically presumed brain death because of flat line EEG and loss of auditory brainstem response (ABR). Among 10 patients, nasopharyngeal lead EEG showed low voltage and slow waves of 10-20 microV in 4 cases and flat line EEG in 6 cases. However, Automatic EEG Analysis system-assisted analysis of EEGs recorded on a magnetic tape revealed that equivalent electric potentials in the scalp EEG from these patients were smaller than those of healthy volunteers, but those in the nasopharyngeal EEG from all patients showed low voltage within the range of distribution in normal volunteers (delta:6, theta 1:6, theta 2:2, alpha 1:1, alpha 3:10, beta 1:6 in 10 cases). The present results show that in addition to the routine scalp EEG, it is clinically useful to employ simple and easy nasopharyngeal lead EEG method and to analyze data using Automatic EEG Analysis System for the judgement of brain death.

Comparison of EEG between nasopharyngeal and scalp leads with the aid of an automatic EEG analysis system.

We analyzed the EEG quantitatively with an automatic EEG analysis system, and introduced this system for investigation the relationship between the scalp EEG and nasopharyngeal EEG. The EEG was recorded from healthy volunteers by photic stimulation. We found that components of slow waves like delta and theta were more rich but alpha component was less in the nasopharyngeal EEG than in the scalp EEG. In the analysis of the rate of the appearance of each EEG wave using photic stimulus, a decrease of alpha 1 and an increase of theta 1 are observed in the scalp EEG while there are no specific changes of them in the nasopharyngeal and scalp EEGs. The nasopharyngeal EEG obtained from the vicinity of the base of the brain was thought to originate from the vicinity of the brain stem.

[A simplified method for evaluation of brain function--basic and clinical study on electroencephalogram by the nasopharyngeal lead].

In making a decision for death, it is well said that brain stem electroencephalogram (EEG) could be an important index. We applied Ishida's nasopharyngeal lead to animals (canine), normal healthy men and patients with brain dysfunction. In normal healthy men, the nasopharyngeal lead revealed clear brain waves. Additionally, we tried a photic stimulation in all the models and a gradual removal of the cerebrum in canines to investigate whether interaction between Ishida's nasopharyngeal lead and the cortical electroencephalogram exists. Furthermore, whenever there was a chance, we examined instances with brain dysfunction by the method, and confirmed that brain waves obtained from Ishida's nasopharyngeal lead were derived from the skull base.