A randomised controlled trial of pectoral nerve-2 (PECS 2) block vs. serratus plane block for chronic pain after mastectomy.

Thoracic interfascial plane blocks are effective for post-mastectomy acute analgesia. However, their effects on chronic pain are uncertain. We randomly allocated 80 women equally to pectoral nerve-2 (PECS 2) block or serratus plane block. The pectoral nerve-2 block reduced the rate of moderate or severe chronic pain from 13/40 (33%) with the serratus plane block to 4/40 (10%), p = 0.03, adjusted odds ratio (95%CI) 0.23 (0.07-0.80), p = 0.02. The rates of pain-free women at six postoperative months were indeterminate, 10/40 (25%) after serratus plane block vs. 19/40 (48%) after pectoral nerve-2 block, p = 0.06, adjusted odds ratio (95%CI) 2.9 (1.1-7.5), p = 0.03. Health-related quality of life at six postoperative months was similar after serratus plane and pectoral nerve-2 blocks, mean (SD) EQ-5D-3L scores 0.87 (0.15) vs. 0.91 (0.14), respectively, p = 0.21. The pectoral nerve-2 block reduced median (IQR [range]) morphine consumption in the first 24 postoperative hours from 6 (3-9 [1-25]) mg to 4 (2-7 [0-37]) mg, p = 0.04. However, acute pain scores after serratus plane and pectoral nerve-2 blocks were similar, median (IQR [range]) 23 (11-35 [0-70]) mm vs. 18 (11-27 [0-61]) mm, respectively, p = 0.44. Pectoral nerve-2 block reduced chronic pain 6 months after mastectomy compared with serratus plane block.

Evaluation of prostate-specific antigen (PSA) membrane test for forensic examination of semen.

Prostate-specific antigen (PSA) is a glycoprotein produced by the prostatic gland and functions to liquefy the seminal fluid. Recently several PSA membrane test kits utilizing an immunochromatographic assay are commercially available for clinical screening of PSA concentration in serum. In this study, we applied the three PSA test kits to forensic examination of semen, and evaluated the sensitivity and specificity of the PSA test kits. The specificity were the same, but SMITEST PSA Card had the highest sensitivity among the three kits. Furthermore we certified that the SMITEST PSA Card was little affected by heat or contaminants such as other body fluids, reagent of preliminary test for semen stain and spermicides, and the PSA kit was applied to 30 casework samples.

Classification of standard alleles of the MN blood group system.

BACKGROUND AND OBJECTIVES: We had classified the M alleles of the MN blood group system into two subtypes, M(G) (standard M) and M(T), based on a G/T substitution in intron 1 of the glycophorin A (GPA) gene. This study provides further study on nucleotide sequences of M(G), M(T) and N alleles to profile the new allele, M(T). MATERIALS AND METHODS: M(G), M(T) and N alleles of the GPA gene were amplified using GPA gene-specific primers to avoid co-amplification of the genes of glycophorins B and E. Then the 5'-flanking region, exons 1-7 and introns 1-4 of the alleles were analyzed by DNA sequencing. RESULTS: There were 17 nucleotide substitutions and deletions between M(G) (standard M) and N alleles. Ten of the M(T) nucleotides were M(G)-type but the other 7 were N-type. M(T) allele also showed one base change and one deletion that were observed in neither the M(G) nor the N allele. Moreover, we found nucleotide substitutions within each allele, allowing further classification of the alleles. CONCLUSION: By the sequence data of M(G), M(T) and N alleles, the three alleles could be further classified into M101 and M102, M201 and M202, and N101 and N102, respectively.

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.

ABO genotyping by inverse PCR technique.

Inverse PCR technique was applied to type three major alleles (A(1), B and O(1)) of the ABO blood group by simultaneously detecting separated allele-determining nucleotides (the 261st base in exon 6 and the 796th and 803rd nucleotides in exon 7) of the ABO gene. A sequence of about 1.7 kb from exons 6 to 7 of each allele was amplified, both termini of the fragment ligated, and allele-typing performed by the inverse PCR-restriction fragment length polymorphism (IP-RFLP) and allele-specific inverse-PCR (ASIP) methods. For intramolecular ligation, primers for the first PCR were designed to have Acc I-restriction sites within the sequences, and both termini of the 1.7-kb fragment were digested with Acc I. Using the IP-RFLP method, the inverse PCR product was digested with Kpn I, Nla III and Dde I, A(1), B, O(1)-standard (O(A)) and O(1)-variant (O(G)) alleles were detected as 365-bp, 272-bp, 193-bp and 128-bp fragments, respectively. By the ASIP method using four allele-specific primers, 222-bp, 124-bp and 232-bp fragments were amplified from A(1), B and O(1) templates, respectively. These techniques would be applicable to detecting separated polymorphic regions of some other genes.

A case of DNA analysis of ABO blood group variant allele A2.

The ABO phenotype of a bloodstain (B) on a knife that was used as a weapon in an attempted murder case was found to be different from that of the Peruvian victim's blood (AB). Serological analysis showed that the A-antigenicity was much weaker than B antigenicity, suggesting that the victim's phenotype was A(2)B or A(3)B. So, the ABO genotypes of the knife bloodstain and the victim's blood were determined by DNA analysis. The 261st G deletion, specific to the O(1) allele, was not detected in the specimens by restriction fragment length polymorphism analysis. Also, the 871st A, specific to the A(3) allele, was not found by the allele-specific amplification method. Amplified product length polymorphism and direct sequencing methods finally demonstrated that the typical B sequence was found in one allele and a single C deletion in the 1,059th-1,061st C stretch in the other allele, indicating that the ABO phenotype of the bloodstain and victim's blood were A(2)B.

Subzero-temperature liquid-liquid extraction of benzodiazepines for high-performance liquid chromatography.

On the basis of the phenomenon that hydrophilic acetonitrile is separated from the aqueous phase at -20 degrees C, we employed a novel extraction method, "subzero-temperature liquid-liquid extraction", to extract benzodiazepines (estazolam and triazolam) from serum or aqueous solution for liquid chromatography. A 1:1 mixture of acetonitrile and the specimen was cooled at -20 degrees C for 20 min to separate the acetonitrile and aqueous phases. The acetonitrile phase was directly injected into a high-performance liquid chromatograph. Recovery rates of the drugs following the first subzero-temperature liquid-liquid extraction were 50.3 +/- 0.6-54.0 +/- 0.9%, which were lower than those (73.9 +/- 3.3-80.6 +/- 0.6% and 81.6 +/- 4.7-96.1 +/- 2.6%) of the first conventional liquid-liquid extraction using diethyl ether and solid-phase extraction using a Sep-Pak C18 column, respectively. However, three to four repeated subzero-temperature liquid-liquid extractions and conventional liquid-liquid extractions resulted in recovery of almost 100% of the drugs. In the chromatogram of the benzodiazepines recovered from serum by the subzero-temperature extraction, no coextracted component interfered with determination of the drugs. Detection limits of the drugs were 0.02-0.08 microgram/mL, and coefficients of variance were 1.14-2.17% suggesting high reproducibility.

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.

Effects of solvent displacement on sensitivity and specificity of monoclonal antibodies for ABO blood grouping of forensic specimens with an absorption-elution test.

Using commercially-available monoclonal antibodies (Bioclone, Neo Kokusai, Monoclonal Wako, Gamma Clone and Seraclone), ABO blood grouping of forensic specimens such as bloodstains, salivary stains, seminal stains, nails, hair and cerebral dura mater was performed with an absorption-elution test. Salivary stains, seminal stains and nails were not typed correctly using the antibodies other than Bioclone reagents, while precise grouping of bloodstains was performed using most antibodies. When hair and dura mater were tested, all of the antibodies induced weak or non-specific haemagglutination, hence correct grouping was not achieved. When the antibody solvents were displaced with 5-20% bovine serum albumin in saline, human serum of the group AB donor, or serum of chicken, sheep or bovine, titers of the reagents increased 2-8 times. Hair and dura mater were able to be typed using Bioclone reagents after solvent displacement with human AB or sheep serum, whereas displacement with the other solvents enhanced non-specific reactions.

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.

A unique origin and multistep process for the generation of expanded DRPLA triplet repeats.

Dentatorubral and pallidoluysian atrophy (DRPLA) is an autosomal dominant neurodegenerative disorder associated with the expansion of a CAG repeat at chromosome band 12p13. Epidemiological studies have demonstrated an increased prevalence of DRPLA in Japan, although several DRPLA kindreds of non-Japanese ancestry have been identified. To define the molecular basis for this geographic variation in prevalence, we have analyzed haplotypes around the repeat in several different ethnic groups. Two intragenic biallelic polymorphisms distinguished three haplotypes, each of which formed a predominant haplotype found in the three major racial populations. All the expanded repeats of Japanese and Caucasian patients studied were associated with a particular haplotype, which otherwise was associated with longer repeats commonly found in Asians. Our results support a multi-step model for repeat expansion, and suggest that expanded DRPLA repeats may have evolved from an ancient chromosomal haplotype of Asian origin. We also propose that a combination of a highly polymorphic microsatellite marker with relatively stable biallelic markers in a range of PCR amplification is a powerful tool for studies on human genome diversity, which may reveal the ancient human migration and the formation of ethnic groups.

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.

Simultaneous early gastric cancer in identical twins: report of a case.

We report the unusual case of identical male twins developing early gastric cancers that were found almost simultaneously. A 39-year-old man underwent a barium-swallow examination to investigate the cause of right hypochondrial discomfort; the examination revealed evidence of gastric cancer in the upper body of the stomach. A diagnosis of adenocarcinoma was confirmed by endoscopic biopsy, and a total gastrectomy was performed. Subsequent screening of the patient's asymptomatic identical twin revealed gastric cancer in the lower body of the stomach, for which distal gastrectomy with Billroth I reconstruction was performed. The histopathological types of the two cancers were similar and both had infiltrated the submucosa. The relevant etiological factors contributing to the development of gastric adenocarcinoma in these identical twins is discussed, following the case report.

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.