[Anesthetic management for neonates with congenital cystic adenomatoid malformation].

We experienced two cases of congenital cystic adenomatoid malformation (CCAM) diagnosed by prenatal ultrasonography. The first case with type I CCAM underwent the resection of the right upper lobe 8 days after birth. Postoperative course was uneventful. The second case with type III CCAM developed non-immune hydrops, severe mediastinal shift, and polyhydroamnios at 29 weeks' gestation. At 30 weeks' gestation, cesarean section was performed. Immediately after birth, the resection of lung tumor for the neonate was performed. However, the neonate died on the 1st postoperative day.

Polymorphisms in IgG Fc receptor IIB regulatory regions associated with autoimmune susceptibility.

Autoimmune diseases involve multiple genes. While functions of these genes are largely unknown, some may be related to an intrinsic hyperresponsiveness of B cells. B-cell responses are controlled by signaling thresholds through the B-cell antigen receptor (BCR) complex. The B1 isoform of type II IgG Fc receptors (FcgammaRIIB1) is exclusively expressed on B cells and serves as a negative regulator for inhibiting BCR-elicited activation. Thus, its allelic variants associated with functional deficits could be examined for possible associations with susceptibility to autoimmune diseases. We found that there are three types of polymorphisms in the reported FcgammaRIIB transcription regulatory regions in mouse strains. Compared to normal healthy mouse strains (group III), autoimmune disease-prone strains (group I) share three deletion sites: two in the promoter region and one in the third intron. Strains (group II) that per se are not autoimmune-prone, but have potentials to accelerate autoimmune diseases share two deletion sites in the third intron: one identical to that in group I and the other unique to group II. These polymorphisms correlated well with extents of down-regulation of FcgammaRIIB1 expression in germinal-center B cells upon stimulation with antigens and up-regulation of IgG antibody responses. Our data imply that these FcgammaRIIB polymorphisms are selected evolutionarily for natural defense against pathogens, and that such polymorphisms may, in turn, form the basis of one aspect of autoimmune susceptibility.

[Fatal child abuse in Japan and Germany. Comparative retrospective study]. / Tödliche Kindesmisshandlung in Japan und Deutschland. Vergleichende, retrospektive Untersuchung.

In this study a record for comparative international epidemiological studies on autopsy cases of child abuse is introduced. The form was proved in a retrospective comparative survey of cases of fatal child abuse at the Department of Legal Medicine in Kanazawa (Japan) and Institute of Legal Medicine of Lübeck (Germany). A total of 33 cases were included. The following data were evaluated: age and gender of victims and assailants, relationship between victims and assailants, causes and methods of abuse, chief autopsy findings, and causes of death. The results were leading into two directions between Kanazawa and Lübeck: (1) In the years of 1981-1996 in Kanazawa 23 cases of fatal child abuse were autopsied while during the same period in Lübeck only 10 cases were registered. (2) While sexual abuse was not registered in Kanazawa, it was recorded twice in Lübeck.

An autopsy case of suicidal strangulation with four looped rubber bands.

A rare case of suicidal strangulation with four looped rubber bands is reported. A 57-year-old female was found dead in the vestibule of her house, with four looped rubber bands wrapped around her neck. Her face was markedly congestive, and lots of petechial hemorrhages were seen on the face and in the palpebral conjunctive. At the medico-legal autopsy, the ligature mark was 1.7 cm in maximum width at the anterior part of the neck and accompanied by such vital reactions as linear hemorrhage, but there were no other injuries to account for her death. Many petechial hemorrhages were also found in the visceral pleurae and epicardium. There was hemocoagulum in the cardiac blood, and the other organs were as well markedly congestive. Gas-chromatographic analysis revealed ethanol; 0.83 mg/ml in the heart blood and 1.37 mg/ml in the urine. No other drugs or poisons were found with Toxi-Lab and REMEDi-HS systems. In addition, police investigation denied the possibility of homicide and disclosed her living will stating that she had suffered from a chronic heart disease. This case was of suicidal strangulation with looped rubber bands.

False identification of urinary ephedrine as methamphetamine by gas chromatography/mass spectrometry with a DB-17 column.

A 17-year-old motorcyclist was killed in a traffic accident. The urine sample collected at autopsy was screened by the Toxi-Lab system; it showed a possible presence of either methamphetamine (MA) or ephedrine (Eph). Gas chromatography-mass spectrometry (GC-MS) was carried out for a trifluoroacetyl (TFA)-derivatized sample after extraction with an Extrelut column. A peak was detected at the same retention time as that of the authentic Eph by GC-MS with a DB-17 wide-bore column; the mass spectrum of the peak was, however, very similar to that of MA. The presence of Eph and absence of MA were proved by GC-MS using a DB-1 column. The Eph in the urine was further confirmed by high performance liquid chromatography (HPLC)-MS using an ODS column. By the REMEDi HS system, Eph and methylephedrine were determined in the urine, and their concentrations (hydrochloride salt) were 0.752 and 2.13 micrograms/ml, respectively. Ethanol was detected as well in the blood (1.55 mg/ml) and urine (2.01 mg/ml) by a pulse heating method. Nicotine, caffeine and dihydrocodeine were also qualitatively detected in the urine by Toxi-Lab, HPLC-MS and REMEDi HS systems. The present case warned that urinary Eph can be misidentified as MA by GC-MS with an intermediately polar column after TFA derivatization.

Use of REMEDi HS in emergency toxicology for a rapid estimate of drug concentrations in urine, serum, and gastric samples.

The REMEDi HS is a broad spectrum drug identification system, designed for emergency toxicology screening and forensic applications. The total analysis time is about 20 min. The current library has 555 drugs and metabolites. The system has a software routine that uses an internal standard (IS) to perform quantitative analysis for target compounds when calibrators are available; further, response factors (RF) are supplied for a rapid estimate of drug concentrations when calibrators are unavailable. In the present study, The concentrations of six drugs (bromisovalum, ephedrine, hydroxyzine, diphenhydramine, ranitidine, and lidocaine) and a metabolite of lidocaine (glycinexylidide) were determined using both methods. The slopes of the regression lines between the rapid estimate method and the IS method were generally within 20% of unity, in agreement with the manufacturer's claim. Semiquantitative estimates based on RF also showed good agreement with results obtained using multipoint calibration. These estimates were sufficient for clinical differentiation of routine and toxic levels. Our study demonstrated that the REMEDi HS is particularly useful for a rapid estimate of drug concentrations in the samples from emergency cases when calibrators are not readily available. Our study also showed that this system can be used for the therapeutic monitoring of ranitidine, bromisovalum, lidocaine, and diphenhydrmine.

[Death due to cold of a 63-year-old male associated with hepatocellular carcinoma--bibliographic consideration on alcohol metabolism of the patients with liver dysfunction].

In the early morning of late December, a 63-year-old man was found dead in front of his apartment house. At medico-legal autopsy, subcutaneous hemorrhages of the occipital region and linear fractures of the occipital bone was observed, but there was no cerebral contusion leading him to death. Many white tumors, ranging from 0.5 to 2.0 cm in diameter, were observed in the liver. On sections of the liver, hepatic parenchyma was found entirely occupied by the tumors, which were histopathologically diagnosed as moderately- or well-differentiated hepatocellular carcinoma. Alcohol concentrations of the mixed intracardiac blood and the urine were determined to be 1.9 and 3.6 mg/ml, respectively. Through the police investigation, it became clarified that the male had been sleeping in front of his apartment house about at 10:30 p.m. on the day before he was found dead. The cause of his death was, therefore, considered due to cold, and the authors gave a bibliographic consideration on blood and urine alcohol concentrations and alcohol metabolism of a patient with liver dysfunction due to hepatocellular carcinoma.

A case of death due to outbreak of fire during thinner abuse.

A 22-year-old male, who could not escape from a residential fire, was found dead at the scene, although the other three persons who had been in the same room escaped safely. At the medico-legal autopsy, the burned body presented the second degree level of body destruction (T. Nagano. Jpn J Legal Med., 1982) and the pugilistic attitude caused by fire. Soot was observed not only in the trachea but also in the bronchi or even the alveoli. Forensic toxicological analyses were performed using the intratracheal gas, left and right ventricular blood, urine, stomach contents, cerebrum, cerebellum, left and right lungs, liver, spleen, kidney, skeletal muscle and adipose tissue. The carboxyhemoglobin concentration was 61.4% in the left ventricular blood and 59.5% in the right ventricular blood. Ethanol and toluene were qualitatively detected in the intratracheal gas by gas chromatography-mass spectrometry. The ethanol and toluene concentration was 0.48 mg/g and 20.4 microgram/g, respectively, in the blood, and 0.40 mg/g and 28.7 microgram/g, respectively, in the cerebrum. Moreover, the urinary hippuric acid concentration was 1.76 mg/ml. The ethanol concentrations were not at the toxic level, while the toluene concentrations in the blood and cerebrum were almost at the lethal level. However, since the preservation of the capacity for vital reactions was apparent at the autopsy in the form of soot in the air passages and the formation of CO-Hb in the blood, it was surmised that the victim was still alive when the fire broke out, but subject to severe disturbance of the central nervous system function. The cause of death in the present case was diagnosed as death due to fire.

Experimental studies on postmortem diffusion of ethanol-d6 using rats.

In an investigation of postmortem ethanol diffusion deuterium-labeled ethanol-d6 was instilled by peroral gavage immediately after death by CO into the stomach of rat carcasses which were subsequently kept for 12-72 h at 5 or 30 degrees C. The heart blood, abdominal fluid and several tissues were collected and analyzed by head space gas chromatography-mass spectrometry. Rat carcasses showed no macroscopic changes until at least 72 h at 5 degrees C, and 12 h at 30 degrees C. At 30 degrees C, slight macroscopic change was observed after 24 h, moderate change after 48 h and marked change after 72 h. In the abdomen ethanol-d6 diffused gradually into neighboring organs (hepatic left lobe, left kidney and spleen) at 5 degrees C, with ethanol-d6 reaching a peak concentration of 0.75-2.38 mg/g at 24 h. At 30 degrees C, ethanol-d6 was also detected in neighboring organs and reached a peak concentration of 1.06-2.61 mg/g at 12 h. Thereafter, the ethanol-d6 concentration in the liver, kidney and spleen decreased, with concentrations ranging from 0.30 to 0.61 mg/g at 30 degrees C and 0.05 to 1.47 mg/g at 5 degrees C at 48 h. In the femoral skeletal muscle, ethanol-d6 was not detected until 24 h or more storage at 30 degrees C and never detected at 5 degrees C. In the brain and the organs in the thoracic cavity ethanol-d6 was detected after 12 h or more at 5 or 30 degrees C. Comparison of these results of direct peroral gastric instillation with those when ethanol-d6 was injected into the stomach through a laparotomy incision suggest that the brain and thoracic cavity changes were a result of diffusion from the mouth and esophagus. After 24 h at 30 degrees C, the postmortem ethanol production (0.33-0.85 mg/g) was comparable to those in previous reports. These results indicate that the assessment of ethanol concentration in the heart blood and organs in autopsy cases must be carefully conducted in comparison with the alcohol content of the stomach.

Sex determination by polymerase chain reaction on mummies discovered at Taklamakan desert in 1912.

Sex determination was performed by the polymerase chain reaction (PCR) on eight adult mummies and one child mummy which were discovered at Taklamakan desert in 1912 and now belong to the Lüshun Museum in China. Archaeologically, these mummies were humans living in the seventh century, that is, more than 1300 years ago. Putative sex determination was performed based on external morphology for six of the eight adults, but it was impossible for the other two adults and one child mummy due to marked destruction on the external morphology. Hair, muscle and skin samples were then collected from each adult mummy, and skin and rib samples from the child mummy. Forty PCR cycles were performed as follows: denaturation at 94 degrees C for 40 s, annealing at 55 degrees C for 30 s and extension at 72 degrees C for 1 min. The primer and PCR reaction mixture were prepared according to the report by Witt and Erickson (M. Witt and R. P. Erikson, A rapid method for detection of Y-chromosomal DNA from dried blood specimens by the polymerase chain reaction. Hum. Genet., 82 (1989) 271-274)). Two different pairs of primer were used. One was X1, X2 (X1: 5'-AATCATCAAATGGAGATTTG-3'; X2: 5'-GTTCAGCTCTGTGAGTGAAA-3') to flanking the 170 bp fragment of the alphoid repeats on the human X chromosome, and the other was Y11, Y22 (Y11: 5'-ATGATAGAAACGGAAATATG-3'; Y22: 5'-AGTAGAATGCAAAGGGCTC-3') to flanking the 130 bp fragment of the alphoid repeats on the human Y chromosome. Extracted DNA solutions from mummy samples was purified using a spin column (T. Yoshii, K. Tamura, T. Taniguchi, K. Akiyama and I. Ishiyama, Water-soluble eumelanin as a PCR-inhibitor and a simple method for its removal. Jpn. J. Legal Med., 47 (1993) 323-329 (in Japanese with English abstract) for removing PCR-inhibitors, and bovine serum albumin (BSA) was employed to inhibit the remaining impurities even after the purification with the column. In six adult cases where the putative sex was determined from external morphology, the sex in five cases was consistent with that by PCR using hair, muscle, and skin samples, but the other one was inconsistent. In two adult cases where sex estimation was externally impossible, the sex was determined to be male because both X-specific and Y-specific bands were clearly detected. The child mummy was definitely male. This study shows that the sex determination was possible by the PCR method even with very ancient human samples > 1300 years old, that spin column was useful for removing impurities in the DNA solution from ancient human samples and that the BSA of optimum concentration suppressed the action of the PCR-inhibitory factors.

Screening of volatile substances and determination of toluene (a thinner component) in the blood and urine in emergency medical care and autopsy cases by the pulse heating method.

Screening of volatile substances was performed by pulse heating gas chromatography - mass spectrometry (GC-MS) using a GS-Q column in 211 emergency medical care and 342 autopsy cases. At least 36 standard substances could be separately detected. Six kinds of volatile substances were screened in a total of 553 cases. Toluene and/or hippuric acid were detected in the blood and/or urine in respectively, 4 emergency medical care and 8 autopsy cases. There were 11 abusers (9 males and 2 females) in these 12 positive cases. The ages of the abusers ranged from 13-26 years. There was no particular pattern to the monthly frequency distribution of identification of thinner (toluene) abuse cases, which occurred throughout the year. It is believed that these data at least partly reflect the present status of thinner/glue abuse in Japan. We conclude that pulse heating GC-MS is useful in the screening and quantitative determination of volatile substances including toluene and other thinner/glue components.

A new surgical method from the viewpoint of personal identification: a case of identification based on the presence of a metallic Z-stent.

To the best of our knowledge, the first case of successful personal identification based on the characteristic finding of a metallic Z-stent, unexpectedly discovered in the common bile duct, is reported. An unknown human male cadaver was found at the seaside. The age was estimated to be approximately 50-70 years and the postmortem interval was considered to be approximately 2 weeks. There was a small surgical scar of 1.5 cm in diameter in the epigastrium. The cadaver was group A in the ABO blood group system. Internally, the following severe injuries were observed: the dislocation of the occipito-atlantal joint, fractures of the cervical and thoracic portions of the spinal column, multiple fractures of the bilateral ribs, and ruptures of the spleen, left kidney and liver. Furthermore, a 4.5 cm long metallic stent was found in the common bile duct, and the gallbladder was not thoroughly identified because of the severely tight adhesion of the surrounding tissues. In every cut surface of the liver, multiple bile plaques, signifying the presence of long-standing cholestasis, were conspicuous. A microscopic examination revealed the invasion of cancer cells into the liver, which was diagnosed as well or moderately differentiated adenocarcinoma, although the primary origin of the adenocarcinoma could not be determined. The cause of his death was diagnosed as traumatic shock. According to the police investigation after the autopsy, it was revealed that a 60-year-old male, suffering from gallbladder carcinoma, had disappeared.(ABSTRACT TRUNCATED AT 250 WORDS)

Determination of the volatile anesthetics halothane, enflurane, isoflurane, and sevoflurane in biological specimens by pulse-heating GC-MS.

Four kinds of volatile anesthetics (halothane, enflurane, isoflurane, and sevoflurane) that were dissolved in 3 microL of experimental plasma samples were examined by the pulse-heating gas chromatographic-mass spectrometric method, and this approach was found to be reliable for qualitative and quantitative analysis. The analytical results also showed good recovery and accuracy. This method was then applied to real blood specimens taken from patients during surgery. The same blood specimens were also analyzed simultaneously by the conventional headspace method for comparison. The data for the clinical blood specimens examined by these two methods showed reasonable correlation coefficients of 0.914 (enflurane) and 0.937 (sevoflurane). These results indicate that the pulse-heating method is applicable for toxicological and clinical analysis of several kinds of volatile anesthetics.

Toxicological analysis for drugs and poisons using the formalin-fixed organ tissues. 1. Methamphetamine.

Methamphetamine (MA) was analyzed using formalin-fixed organ tissues from experimental animals and an autopsied cadaver. The brain, lung, liver, kidney and skeletal muscles of rabbits that had been administered MA were collected, and then fixed with a non-buffered 10% formalin solution at room temperature (10-20 degrees C) for five different fixation periods (1, 3, 7, 14 and 28 days). After the Extrelut column extraction, MA was derivatized using anhydrous trifluoroacetic acid (TFA) and analyzed by GC-MS using a DB-17 column. MA-TFA in the eluates from the organ tissues after the formalin-fixation could be identified by GC-MS. Even one day after the formalin-fixation, concentrations of MA in the fixed organ tissues already significantly decreased to 1.3-3.1% when compared with those of MA in the non-fixed tissues, and to 0.04-0.4% in all of the fixed organ tissues at 28 days after the fixation. Moreover, MA in the organ eluates from the autopsied male cadaver after one-month formalin-fixation was qualitatively detected. These results obviously show the usefulness of MA detection using formalin-fixed organs in the practice of forensic medicine and police science.

Changes in the alpha-adrenoceptors in the medulla oblongata including nucleus tractus solitarii of spontaneously hypertensive rats.

The nucleus tractus solitarii (NTS) is a brain stem center mediating depression of blood pressure. In order to elucidate a possible mechanism for the central regulation of blood pressure, we studied noradrenergic indices in the medulla oblongata, a region including the NTS, in spontaneously hypertensive rats (SHR) as compared with normotensive controls of the Wistar Kyoto strain (WKY) at 12 weeks of age. The medulla oblongata was the only brain region showing a significantly low noradrenaline level in the SHR as compared with WKY rats; the level is also significantly decreased at 8 weeks of age. The alpha 1-adrenergic binding sites, as measured with 2-(2', 6'-dimethoxy) phenoxyethylamine-methylbenzodioxan [3H]WB4101 showed significant increases in KD and Bmax values in medulla oblongata homogenates from rats of both strains from 4-12 weeks after birth, with no significant interstrain difference. On the other hand, the KD and Bmax of the alpha 2-sites, measured by [3H]yohimbine binding, were reduced in SHR as compared to WKY animals, even at 4 weeks after birth when hypertension was not yet apparent. As expected, the relatively selective alpha 2-antagonist, clonidine, was a potent inhibitor of [3H]yohimbine binding but not of [3H]WB4101 binding in these homogenates. The results suggest that some genetic disorder in the alpha 2-adrenergic transmission system in the NTS region may be involved in the development of hypertension in the SHR rats.