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 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.

[Case of death by fire with kerosene--analysis of contents of trachea and stomach].

UNLABELLED: In death caused by fuel oil burning, it's difficult to examine the vital reaction in the burning skin surface. In these cases, in stead of skin examination, we've been determining the fuel oil in blood. In this case, besides this method, we tried to examine the contents of the trachea of a person who died of kerosene oil burns. CASE: A 49-year-old female was found dead in a cabin. Burns on her body ranged from first to fourth-degree, and 91% of the body was charred. Carbon particles were detected within the trachea and the bronchus, and were slightly detected in the gastric contents and the esophagus. Carboxyhemoglobin concentration was found to be 21% in the right heart blood and 22% in the left heart blood. The level of cyanide detected was 4.3 microM in the right heart blood and 1.7 microM in the left heart blood. Ethanol was not detected in either sample. Kerosene components were detected in each sample (blood, trachea content, gastric content and body surface). According to the formulation of kerosene components, results of contents of the trachea were most likely from a kerosene on the market. In the blood, many volatile paraffin hydrocarbons were found, and, on the body surface, many high boiling-point paraffin hydrocarbons were detected. The means that values of detected kerosene formulation from the blood and trachea contents were similar to types of kerosene on the market. From these results, we concluded that the victim inhaled kerosene vapor.(ABSTRACT TRUNCATED AT 250 WORDS)

[Determination of benzodiazepines by thermospray liquid chromatograph-mass spectrometer. Part 1. Nitrazepam, estazolam, bromazepam, flunitrazepam].

Thermospray liquid chromatography mass spectrometric method is described for the determination of the benzodiazepines (Nitrazepam, Estazolam, Bromazepam, Flunitrazepam). Reversed-phase liquid chromatography was performed using a 15 cm Shim-pack CLC-ODS (Shimadzu) column, with acetonitrile-water (40:60) + 0.1 M ammonium acetate as mobile phase, at a flow rate of 1.0 ml/min. The temperature of the vaporizer, block and TH of the source block were 166, 270 and 275 degrees C, respectively. Positive ion thermospray mass spectra by thermospray ionization (TSP ionization) mode or thermospray on filament ionization (filament-on ionization) mode were obtained. Formation of the MH+ ion was observed as a base peak under TSP ionization and filament-on ionization conditions and fragment ions were very few. On both ionization mode, peaks representing nitrazepam as MH+ at m/z 282 at a retention time (R.T.) of 6.4 min, from estazolam as MH+ at m/z 295 at an R.T. of 6.4 min, from bromazepam as MH+ at m/z 316 at an R.T. of 4.5 min and from flunitrazepam as MH+ at m/z 314 at an R.T. of 8.8 min. The detection limit for all the benzodiazepines under investigation was less than 0.5 ng (S/N = 9.4 +/- 4.6) using selected ion monitoring.

[Study on the histochemical staining of boric acid].

The detection of boric acid in the tissue is of significance in investigating its toxicity. Because of this, we have devised a histochemical staining method to detect the presence of boric acid. The outline of this method follows. Frozen 12-14 microns sections, cut by a cryostat, are fixed in anhydrous ethanol and stained for 20 minutes in a protonated curcumin solution. Washing in acetic acid follows, and a red stain results if boric acid is present. This method causes a reaction, in which rosocyanin is formed by the reaction of boric acid and the protonated curcumin, and this principle is now used when an analysis of boric acid is needed. As to procedure, a 1 N concentration of sodium hydroxide is dropped onto a part of the stain to be tested, and the presence of rosocyanin is confirmed if the stain turns blue. Consequently, this staining confirms the presence of boric acid.

[Determination of boric acid in biological materials by curcuma paper].

The field of legal medicine has seen a recent increase of poisoning by boric acid and, in cases of emergency, a simple method of making a qualitative analysis of the boric acid content is a necessity. Thus, we have examined curcuma paper (turmeric paper) to see if it can provide a qualitative analysis of the boric acid content in biological materials, so as to identify cases of poisoning. It was found that curcuma paper can provide a preliminary analysis of the quantitative content of boric acid, and that about 0.1 mg/ml of boric acid can be determined. The steps for this testing method follow. First, either blood or urine is acidified with a 6 N concentration of hydrochloric acid, i.e., in the case of urine, 0.5 ml of urine is added to 0.1 ml hydrochloric acid, and for blood, 0.5 ml of blood is added to 0.2 ml hydrochloric acid. If the sample is not sufficiently acidic, more hydrochloric acid is added. Next, a drop of the sample is placed on the curcuma paper and, after drying at room temperature, a red stain results if boric acid is present. (Rosocyanin is formed by the reaction of boric acid and protonated curcumin). Then, a 1 N concentration of sodium hydroxide is dropped onto the stained place, and if rosocyanin is present, the stain will turn blue. Informatively, to make curcuma paper, filter paper (No. 2) is soaked in a saturated curcumin/ethanol solution and then air dried.(ABSTRACT TRUNCATED AT 250 WORDS)