Blood, bone marrow and eye fluid ethanol concentrations in putrefied rabbits

The effect of putrefaction on postmortem blood, bone marrow and eye fluid ethanol levels was evaluated in rabbits. Control and dosed animals were sacrificed and stored at either room temperature (approx. 19 degrees C) or cold temperature (approx. 3.5 degrees C) for as long as 28 days. Control animals stored at room temperature showed ethanol levels in the bone marrow that peaked at 7 days after sacrifice, followed by decreases to a nondetectable level at 21 days. Overall decreases were demonstrated in bone marrow of dosed rabbits stored at room temperature for all postmortem intervals. The control animals stored at low temperature showed no ethanol in the bone marrow and blood until 21 days after sacrifice. Dosed rabbits stored at low temperature showed no significant changes in blood and marrow ethanol until 21 days after sacrifice.     

Blood versus bone marrow pentobarbital concentrations

Postmortem pentobarbital levels in rabbit heart blood and bone marrow were determined and compared. The average ratio of femur marrow/blood pentobarbital concentrations in 24 rabbits was 1.06 +/- 0.05. The average percent difference between actual plasma pentobarbital concentrations and calculated plasma pentobarbital concentrations was 5.82 +/- 1.96. Concentrations were determined by gas chromatography of extracted, derivatized pentobarbital.     

Experimental studies on death by fire in automobiles and exhaust gas poisoning

Studies were made on the acid-base balance, blood gases, and carbon monoxide (CO), cyanide, and sulfur dioxide concentrations in the blood of albino rabbits that died from automobile exhaust gas poisoning (group I) or fires in cars (complete combustion, group II; incomplete combustion, group III). In group I, the temperature and CO concentration increased gradually to 35 degrees C and 5.2% in 70 min. The animals died after 9 min, when the values were 20 degrees C and 5.2%, respectively. In group II the animals died after 9 min, when the values were 55 degrees C and 1.95%, respectively. In group III, the temperature was very high (870 degrees C), but the CO concentration was not (0.6-1.3%) after 4 min. The animals died after 5 min. In all experimental groups, marked acidosis and hypoxemia were seen, but the CO2 tension (PCO2) was high, in contrast to previous studies on pure CO poisoning. In group I, the level of carboxyhemoglobin (CO-Hb) was significantly higher (91.2 +/- 3.4% in arterial blood, 87.5 +/- 8.1% in venous blood; p less than 0.01) than in groups II and III. Although the O2 tensions of venous and arterial blood (PvO2, PaO2) were very low, that of arterial blood was higher, suggesting that O2 was still being utilized in the tissues at the time of death. In group II, CO-Hb was high (57.7 +/- 16.0% in arterial blood, 61.2 +/- 20.6% in venous blood) and the acid-base balance indicated marked acidosis. In group III, the CO-Hb, PCO2 and cyanide levels in the blood were very high.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparative study of ethanol levels in blood versus bone marrow,vitreous humor,bile and urine

Post-mortem ethanol levels in blood were compared to corresponding levels in rib bone marrow, vitreous humor, urine and bile. In forensic toxicology, a good correlation between blood and a tissue or body fluid is needed to estimate a blood alcohol concentration when blood is unavailable or contaminated. In this study, direct injection and headspace gas-chromatographic techniques were employed to quantitate the ethanol concentrations. Comparable findings by these two techniques showed a reproducibility of results. When the determined bone marrow ethanol levels were corrected for the lipid fraction, a consistent correlation could be established between ethanol levels in blood and bone marrow. The relationship (linearity and ratio range) between ethanol levels in blood and corrected levels in bone marrow was better than that between blood and vitreous humor, bile or urine. This study showed that blood ethanol levels can be predicted by extrapolating the corrected rib bone marrow ethanol level.     

Effect of methamphetamine abuse on the bone quality of the calcaneus.

BACKGROUND: Methamphetamine, a derivative of amphetamine, has been said to cause mainly mental dependency in humans, but only little is known about its physical dependency. Like other narcotics, it may have chronic effects on the human body, so, this study was planned to evaluate it by examining the bone quality of the skeletal system. METHOD: Among the convicts serving in Fuchu Prison, two groups of people were chosen according to their methamphetamine experience. The bone quality of the calcaneus of both abusers (n = 59, ages 41 +/- 11 years) and controls (n = 50, aged 45 +/- 13 years) was examined with an Achilles ultrasound bone densitometer. SOS (speed of sound) and BUA (broadband ultrasound attenuation), both of which were obtained from the measurement and are an indicator of the strength of bone, were compared between the two groups. RESULTS: The SOS of the abuser group was 1559 +/- 24 (mean +/- SD) m/s and this was significantly lower than that of the control group, 1570 +/- 27 (mean +/- SD) m/s (p = 0.017). The BUAs of the abuser group and the control group were 108 +/- 10 and 110 +/- 10 (mean +/- SD) dB/MHz, respectively, and there was no significant difference between them (p = 0.181). CONCLUSION: There was loss of SOS of the calcaneus in methamphetamine abusers.     

Morphine and 6-acetylmorphine concentrations in blood, brain, spinal cord, bone marrow and bone after lethal acute or chronic diacetylmorphine administration to mice

The aim of this study was to evaluate postmortem incorporation of opiates in bone and bone marrow after diacetylmorphine (heroin) administration to mice. Mice were given acute (lethal dose of 300 mg/kg) or chronic (10 and 20 mg/kg/24 h for 20 days) intraperitoneal administration of diacetylmorphine. The two metabolites of diacetylmorphine, 6-acetylmorphine (6-AM) and morphine, were extracted from whole blood, brain, spinal cord, bone marrow and bone (after hydrolysis) using a liquid/liquid method. Quantification was performed by gas chromatography-mass spectrometry (GC/MS). Results showed that after acute administration, opiates were present in all studied tissues. Morphine concentrations appeared to be higher than those of 6-AM in blood (52.4 microg/mL versus 27.7 microg/mL, n=12), bone marrow (87.8 ng/mg versus 8.9 ng/mg, n=6) and bone (0.85 ng/mg versus 0.43 ng/mg, n=6), but 6-AM concentrations were higher than those of morphine in brain (14.0 ng/mg versus 7.4 ng/mg, n=12) and spinal cord (27.8 ng/mg versus 20.8 ng/mg, n=12). No correlation was found for both compounds between blood concentrations and either brain, spinal cord, bone or bone marrow concentrations while a significant one was found between brain and spinal cord concentrations either for morphine (r=0.89, n=12, p<0.001) or 6-AM (r=0.93, n=12, p<0.001), the concentration being higher in spinal cord than in brain. When bones were stored for 2 months, only 6-AM remained in bone marrow but not in bone. After chronic administration, mice being sacrificed by cervical dislocation 24 h after the last injection, no opiate was detected in any studied tissues. Further studies are required, in particular in human bones, but these results seem to show that 6-AM could be detect in bone marrow several weeks after the death and could be an alternative tissue for forensic toxicologist to detect a fatal diacetylmorphine overdose, even if no correlation between blood and bone marrow was observed. On the other hand, neither bone tissue nor bone marrow will allow the confirmation of a chronic diacetylmorphine use.     

Determination of morphine in postmortem rabbit bone marrow and comparison with blood morphine concentrations

The aim of this study is to predict how long after time of death a buried body could be analyzed for opiates in soft tissues and to show the accessibility and suitability of bone marrow as a useful toxicological specimen from buried bodies. Morphine solutions were injected in nine albino rabbits. Doses ranged from 0.3 to 1.1 mg/kg with 0.1 mg/kg increments. One hour after the injections, the rabbits were sacrificed. Blood, urine and bone marrow samples were collected for analysis. After the whole bodies were buried, femur bone marrow specimens were collected on the seventh and fourteenth days. CEDIA was used to monitor morphine contents of the collected samples. All experimental cases showed that the increase in the given morphine doses correlated with the increase in blood and bone marrow morphine concentrations. High morphine concentrations were detected in urine samples, but there was no correlation between the urine and blood or urine and bone marrow morphine concentrations. Statistically meaningful increases in bone marrow morphine concentrations were found parallel to increase of blood morphine concentrations. Seventh and fourteenth day postmortem morphine concentrations also followed this correlation. Morphine concentrations in bone marrow at 7 and 14 day postmortem decreased consistently when compared with bone marrow morphine concentrations collected immediately after death. We conclude that in sudden death when other specimens are unavailable due to degradation, bone marrow can be a most useful specimen. Further experimental research in this area is required to validate bone marrow as an alternative tissue.     

A large-scale study of the relationship between blood and breath alcohol concentrations in New Zealand drinking drivers

Blood alcohol concentrations (BAC) and corresponding breath alcohol concentrations (BrAC) were determined for 21,582 drivers apprehended by New Zealand police. BAC was measured using headspace gas chromatography, and BrAC was determined with Intoxilyzer 5000 or Seres Ethylometre infrared analysers. The delay (DEL) between breath testing and blood sampling ranged from 0.03 to 5.4 h. BAC/BrAC ratios were calculated before and after BAC values were corrected for DEL using 19 mg/dL/h as an estimate of the blood alcohol clearance rate. Calculations were performed for single and duplicate breath samples obtained using the Intoxilyzer (groups I-1 and I-2) and Seres devices (groups S-1 and S-2). Before correction for DEL, BAC/BrAC ratios for groups I-1, I-2, S-1, and S-2 were (mean+/-SD) 2320+/-260, 2180+/-242, 2330+/-276, and 2250+/-259, respectively. After BAC values were adjusted for DEL, BAC/BrAC ratios for these groups were (mean+/-SD) 2510+/-256, 2370+/-240, 2520+/-280, and 2440+/-260, respectively. Our results indicate that in New Zealand the mean BAC/BrAC ratio is 19-26% higher than the ratio of the respective legal limits (2000).     

Blood versus bone marrow isopropanol concentrations in rabbits

The use of bone marrow to determine the blood isopropanol concentrations becomes important when a blood specimen is contaminated or unavailable. The blood/marrow isopropanol ratios were determined in rabbits autopsied 0, 4, and 24 h after sacrifice. The lipid content of the individual marrow specimens was shown to have a significant influence on the range of ratios. When the determined marrow isopropanol concentrations were corrected for lipid content, a better correlation between blood and marrow concentrations was obtained. The ratio (1.45 ± 0.17) was not altered significantly by postmortem time or temperature.Although acetone was not exogenously administered to the rabbits, but rather was endogenously produced from isopropanol metabolism, the relationship between blood and marrow acetone concentrations was somewhat linear. However, the range of observed and corrected blood/marrow acetone ratios was altered significantly by storage temperature, and delays between death and analysis. Thus, under the experimental conditions of this study, marrow isopropanol concentrations may be used to predict blood isopropanol concentrations, whereas marrow acetone concentrations can not.     

A comparative study of ethchlorvynol levels in blood versus bone marrow

Bone marrow may be utilized as an alternative biological sample in cases where uncontaminated blood samples are not available for analyses. Bone marrow/blood ratios of ethchlorvynol as a function of time and dosage level were determined in 40 rabbits. A modified quantitative analysis that produced accurate and reproducible results was employed for the determination of ethchlorvynol levels. Further, ten blood and bone marrow samples containing ethchlorvynol were chosen to study the effects of storage for a period of 24 hours. Studies of blood and bone marrow ethchlorvynol levels with time showed no linear relationship. Bone marrow/blood ratios as a function of dose resulted in close mean averages with a wide range of values. Significant losses in both blood and bone marrow ethchlorvynol levels were evidenced in most of the samples subjected to the 24-hour storage study.     

Quantitative assessment of the percent fat in domestic animal bone marrow

Measurement of the amount of fat in femoral bone marrow can provide a quantitative assessment of the nutritional status of an individual animal. An analytical method is presented for quantitating the percent fat in bone marrow from three domestic species: bovine, canine, and equine. In this procedure, fat is extracted from bone marrow using pentane, and the percent fat recovered is determined gravimetrically. Based on analyses from adult animals (normal body condition scores), the average percentage of fat in the bone marrow was >80%. In cases in which animals have been diagnosed as emaciated or exhibit serous atrophy of fat (body scores of 1 or 2), the femoral bone marrow fat was less than 20%. In domestic animals, bone marrow fat analysis can be a useful, quantitative measure that, when used in conjunction with all other data available, can support a diagnosis of starvation or malnutrition.     

A blood cyanide distribution study in the rabbits intoxicated by oral route and by inhalation

Blood cyanide concentration was determined in rabbits intoxicated orally or by inhalation. Experiments were carried out under urethane anaesthesia. In the inhalation experiments, rabbits inhaled a combustion product containing HCN via the tracheal cannula and in the oral studies animals were administered NaCN solution into the stomach. In addition to the carotid artery and jugular vein blood samples, postmortem samples were obtained from both sides of the heart and the descending vena cava. The arterial cyanide concentration in the inhalation group showed a close relationship with ventilation. After an initial rise, blood levels decreased a little, in some cases with transient apnea. At the last stage it again increased with gasping, reaching its maximal value. After ultimate apnea, the blood cyanide concentration declined. The blood cyanide values were higher in the oral group than in the inhalation group. The difference between the two groups became larger in the inferior order, the left heart blood--the right heart blood--blood in the descending vena cava. The left heart/right heart ratio of the inhalation group was significantly higher than that of the oral group (1.28+/- 0.28 vs. 0.95+/- 0.09). The coefficient of variation (c.v.) of the inhalation group was larger than that of the other group. Within the inhalation group, the left heart blood showed the largest c.v. values and this was probably due to redistribution of the cyanide by bloodstream after attainment of the maximal concentration.     

Distribution of ethyl glucuronide in rib bone marrow, other tissues and body liquids as proof of alcohol consumption before death

Postmortem ethyl glucuronide (EtG) concentrations in rib bone marrow, liver, muscle, fat tissue, urine, blood and bile have been determined by LC-MS/MS. Samples have been taken from twelve corpses during autopsies. In nine corpses EtG could be detected, corresponding blood ethanol concentrations (BAC) were 0.04-0.37 g%. In three cases, no EtG was found; two of these cases showed postmortem BACs - possibly due to putrefaction - of 0.01 and 0.1g%. In rib bone marrow, which is easily accessible during autopsy, EtG concentrations (0.77-9.36 microg/g) have been lower than in blood (2.24-20.46 microg/mL) in eight of nine cases and comparable or higher than in muscle tissue. Therefore, rib bone marrow has been found suitable as matrix for EtG determination. The highest EtG concentrations have been found in urine in all but one case, where the resorption of ethanol had been incomplete. Second highest EtG concentrations have been detected in liver samples. In two cases with putrefaction, EtG could not be detected. In these cases, the detectable ethanol might have been produced partially or in total by postmortem fermentation. However, instability of EtG during putrefaction cannot be totally excluded which might result in a total loss of EtG.     

Blood versus bone marrow methanol concentrations in rabbits

Postmortem methanol levels in bone marrow and heart blood were determined in rabbits. The average ratio of heart blood concentration to observed bone marrow concentration in 36 rabbits was 2.6 ± 0.6 with a range of 1.5 to 4.2. Correcting for the lipid content of the bone marrow decreased the average ratio, reduced the ratio range and improved the correlation. The heart blood to corrected bone marrow ratio was 1.6 ± 0.3 with a range of 1.2 to 2.9. Direct injection gas chromatographic techniques were employed to quantitate methanol concentrations.     

Evaluation of post-mortem ethanol concentrations in pericardial fluid and bone marrow aspirate

This study confirmed post-mortem ethanol concentrations in pericardial fluid and bone marrow aspirate in comparison with those in the blood in medicolegal autopsy cases (n = 140, within 48 h post-mortem). The specimens were examined by head-space gas chromatography/mass spectrometry. Ethanol concentrations in the pericardial fluid (y) were approximately equivalent to those in peripheral blood (x): y = 0.99x + 0.02, n = 44, r = 0.972. A high stomach ethanol concentration (>10 mg/ml) appeared to mildly affect the pericardial levels. There was no significant interference in drowning cases. Ethanol concentrations in bone marrow aspirates (y) also showed a good correlation with those in the peripheral blood (x): y = 0.77 x + 0.02, n = 20, r = 0.981. A dissociation was observed in cases of delayed death from hemorrhagic/traumatic shock and elderly victims. These findings suggest that pericardial fluid and bone marrow aspirate can be used as an alternative material when adequate blood specimens are not available.     

Influence of blood loss on the pharmacokinetics of citalopram

Extended blood loss results in several compensatory physiological mechanisms, including transfer of extravascular fluid into the blood circulation. If drugs are present in the body, this fluid exchange may imply that blood drug concentrations found in a trauma victim may differ from the concentrations present at the time of the trauma. To address this issue, an animal model was used to investigate the influence of blood loss on pre-existing levels of the antidepressant drug citalopram and its demethylated metabolites. Rats were administered citalopram either acutely (40 mg/kg, orally) or chronically (20 mg/kg daily, subcutaneously) for 6 days using osmotic pumps. In the experimental rats, blood loss was accomplished by withdrawing 0.8 mL blood at 10 min intervals during 70 min. In the control rats, blood was withdrawn at 0 and 70 min only. Blood, brain and lung drug concentrations were analyzed with an enantioselective HPLC method. In the chronically treated rats, the ratios between final and initial citalopram concentrations were 1.08 +/- 0.15 and 1.01 +/- 0.09 in the experimental rats and controls, respectively, indicating no major effect of blood loss. In contrast, acute oral administration resulted in increased ratios in the exsanguinated rats as compared to controls (1.84 +/- 0.50 versus 0.73 +/- 0.07; p = 0.0495). In conclusion, the observation of increased blood drug levels in the acute oral rats indicates that absorption of fluid from the gastrointestinal tract may be important in the intravascular refill. Further, in the interpretation of post-mortem blood levels of drugs, these physiological mechanisms should be taken into account.     

A study on the combined action of CO and HCN in terms of concentration-time products

Acute toxicity at single and combined exposures of CO and HCN was studied on rats in terms of concentration-time product (ppm . min) necessary to kill animals (lethal CT). The animal was exposed individually to test gas in an animal chamber made of transparent plastics, and test gas was made in gas chamber connected to the animal chamber by a wide and short piece of plastic tube. HCN was produced by addition of NaCN solution to H2SO4 and in case of CO exposure, various amounts of pure CO were introduced. During exposure, gas samples were frequently taken. After exposure, blood sample was withdrawn from the right side of the heart. CO concentrations in the gas and blood were determined gas chromatographically. HCN in the gas sample was measured spectrophotometrically, after being absorbed into NaOH solution in a glass vessel devised by our laboratory. At single exposures, mean lethal CT for CO was 78,000 +/- 22,000 and for HCN was 4,700 +/- 940. In combined exposure, various combinations of CO and HCN were used. A fractional CT, defined as a ratio of CT to lethal CT, multiplied by 100, was calculated for each gas. A linear relationship between fractional CTs of HCN and CO was considered to show a simple additive action between the two gases. The sum of both fractional CTs averaged 100 +/- 26. On the other hand, linear relation was not observed between blood levels of the two toxicants at death.     

Iliac cancellous bone in drug addicts: a histomorphometric study

Histomorphometry was used to determine structural bone changes in drug addicts. Iliac crest bone biopsies were obtained at autopsy from 28 subjects (21 male, 7 female, aged 18 to 45 years) who had a history of drug abuse and had died due to overdose of illicit drugs. For histomorphometry, undecalcified sections were investigated using the Merz grid. The following histomorphometric indices were measured and calculated: BV/TV, BS/BV, Tb.Th, Tb.N, Tb.Sp, OV/TV, OS/BS, Ob.S/BS, O.Th, ES/BS, Oc.S/BS, and N.Oc/T.A. In 28 controls (24 male, 4 female, aged 17 to 47 years) trabecular bone specimens were investigated in the same way. The parameters in drug addicts did not show any correlation to age, body weight, height or sex differences. Trabecular bone volume and trabecular thickness were slightly but not significantly increased (BV/TV: 23.37 +/- 5.77% (mean, SD), controls 22.23 +/- 5.08%, p = 0.434; Tb.Th: 172.67 +/- 36.83 mcm, controls 169.73 +/- 36.13 mcm, p = 0.764). Only the eroded surface was significantly different to the controls (ES/BS: 8.16 +/- 2.04%, controls 6.96 +/- 2.17%, p = 0.038). We conclude that the incidence of metabolic bone disease in drug addicts is low.     

The effect of ibuprofen on ethanol concentration and elimination rate.

Pursuant to a recent driving under the influence (DUI) case, a medical study of six subjects was cited reporting that ibuprofen causes a decrease in the maximum rate of elimination of ethanol. Such a drug interaction is of significant forensic science interest and warrants further examination. This study investigates the effect of ibuprofen on ethanol elimination rate and ethanol concentration in nineteen volunteers. Volunteer subjects were randomly assigned to two groups administered either a placebo followed by ethanol or ibuprofen followed by ethanol. Subjects served as their own control. Blood ethanol concentrations were monitored every 30 to 60 min for up to 4 h with Intoximeter 3000 instruments. A blood sample was drawn at the final Intoximeter test and analyzed for ethanol and ibuprofen by gas chromatography and mass spectrometry, respectively. The mean elimination rate (+/- SD) as calculated using Widmark's elimination factor was 0.018 +/- 0.006 g/dL for ethanol and 0.017 +/- 0.007 g/dL/h for ethanol with ibuprofen. Mean ethanol concentrations (g/dL +/- SD) were: 0.095 +/- 0.026 (ethanol) and 0.095 +/- 0.033 (ethanol and ibuprofen) at 30 min; 0.077 +/- 0.026 (ethanol) and 0.075 +/- 0.031 (ethanol and ibuprofen) at 150 min; and 0.089 +/- 0.025 (ethanol) and 0.087 +/- 0.030 (ethanol and ibuprofen) overall. There was no statistically significant affect of ibuprofen on either the peak blood ethanol concentration or the ethanol elimination rate (p less than or equal to 0.001). These results reveal no evidence of a significant ethanol-ibuprofen interaction.     

Human identification and sex determination of dental pulp, bone marrow and blood stains with a recombinant DNA probe

Recombinant DNA hybridizing specifically to a 300 nucleotide repeat DNA sequence (BLUR8) of human specificity and to human repeat DNA sequence (pHY10) on the Y chromosome was used for human identification and sex determination of degraded DNA samples of blood stains, dental pulp, and bone marrow. This radioactive technique enabled reliable and sensitive human and sex determination from blood stains that were more than 80 years old. Less than 1 piece of 0.5 cm length thread of blood stain was enough for both tests. DNA from relatively fresh dental pulp and bone marrow was clearly identified. The human identification test, which could recognize up to 0.3 ng DNA correctly, was 3 to 5 times more sensitive than the sex determination test.