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Cases of fatal triazolam poisoning.

Benzodiazepines have been regarded as relatively safe drugs but since triazolam was introduced into Denmark in 1978, six fatal cases of poisoning by triazolam alone or in combination with ethanol, have occurred in East Denmark. In one case where no ethanol was detected, a blood triazolam concentration of 0.11 mg/l was found. In another case in which blood was unavailable because of putrefaction, muscle was analyzed. In the four cases in which the ethanol concentration was in the range 110-202 mg/100 ml, the triazolam concentration range was 0.04-0.22 mg/l. Blood concentrations from these five cases were compared to those from living persons who had also ingested the drug, and these were in the range 0.002-0.03 mg/l (n = 28), with an average concentration of 0.01 mg/l.

Adult↗

Gas--liquid chromatographic retention indices of 296 non-drug substances on SE-30 or OV-1 likely to be encountered in toxicological analyses.

The advent of the widespread use of selective detectors (electron capture detector, phosphorus/nitrogen dectector) for gas--liquid chromatography used in toxicological analyses has revealed the presence of hitherto unseen interfering materials. These substances may be conveniently grouped into (1), anti-oxidants; (2), putrefactive and endogenous compounds; (3), pesticides; (4), food additives, flavours and fragrances; (5) plasticisers, plastic additives and vulcanising agents and (6), scintillation reagents. To facilitate the identification of these materials, retention indices on the dimethyl silicone phases SE-30 or OV-1 have been compiled by the two laboratories to include 296 such compounds. Most gave single peaks, but some gave complex patterns indicating that they were mixtures of compounds. Of the 296 compounds, 14 did not give observable peaks, 8 gave 2 or 3 peaks and 44 gave more than 3 peaks. To determine the interlaboratory difference between retention index measurements, 17 compounds were chromatographed by both laboratories: the mean difference was +/- 13 retention index units with only one greater than +/- 50 retention index units. Examples of how these materials may be encountered during toxicological analyses are given. Data are also presented on compounds which have been used as internal standards.

Chromatography, Gas↗

Histomorphologic studies of the glottic region to elucidate the pathogenesis and functional significance of fibrinoid vocal cord lesions in infants and young children.

Focal lesions of the vocal cord in form of fibrinoid necrosis and thickening of the basement membrane are morphological findings in infants and young children that receive only scant attention and are also controversially valued. We discuss two causes for the pathogenesis of these findings: (1) recurrent laryngospasms and (2) a laryngeal phonotrauma resulting from frequent spells of shrill, presumably high-frequency crying. The aim of our study therefore was to gain insight into the development and functional significance of these lesions by examining their histological patterns and to draw the pertinent conclusions. In 56 deaths (natural and unnatural causes, fetal life up to 4 years, no pathological or traumatic laryngeal lesions) histological investigations were carried out on putrefaction-free material. For the histomorphological studies, horizontal sections through the region of the vocal cord were obtained and stained according to conventional and immunohistochemical staining methods. In 42 cases the thickness of the basement membrane was determined morphometrically. In 12 of the 56 cases (21%) we observed staged variations of focal lesions, ranging from fibrinoid necrosis to degenerative lesions with pronounced macrophage involvement and deposits of PAS-positive plaques. Measurements at the basement membrane revealed a thickness of 7.9 microm. Separate measurements at the anterior and posterior portions of the vocal fold showed a ratio of 1.7:1. When the cases were subdivided into those with and without fibrinoid necrosis the ratios were 2.5:1 and 1.5:1. This different pattern makes it unlikely that laryngospasm is the sole or essential cause. Pathogenetically more important might be phonatory overstraining due to maximum vibrations in only the anterior portions of the vocal fold, which is not specific for later SIDS victims. This assumption is also supported by the often observed conspicuous phonetic behavior of some later SIDS victims and the absence of these stage-like vocal cord lesions in stillborns and adults. It is presumably an age-specific, temporary finding which is completely reversible.

Basement Membrane↗

DNA typing of human remains found in damp environments.

DNA typing is often used to determine identity from human remains. The environment to which the material has been exposed, however, is crucial for the success of the investigation. Damp conditions in particular can cause a rapid degradation of DNA, even in bone and teeth, and thus reduce the chances of successful typing. Here, we present the results of investigations performed on four skulls that had been lying in a damp environment for periods ranging from almost 1 year to about 45 years. In none of these cases was DNA typing successful on bone or, where present, on teeth. Where remnants of brain tissue were used, however, complete STR typing was possible in one case, partial STR typing in another, and mtDNA sequencing could be carried out in three cases. These findings suggest that brain tissue is relatively resistant to putrefaction in damp environments and, unlike bone, appears to exhibit a certain degree of protection against DNA degradation.

Aged↗

Postmortem diffusion of drugs from the bladder into femoral venous blood.

We describe significantly elevated drug concentrations in the femoral venous blood due probably to postmortem diffusion from the bladder. A 16-year-old deceased male was found in a shallow ditch in winter. The estimated postmortem interval was 9 days and putrefaction was not advanced. The cardiac chambers contained fluid and coagulated blood and a small amount of buffy coat clots. Diffused hemorrhages were found in the gastric mucosa. The bladder contained approximately 600 ml of clear urine. Gas chromatographic-mass spectrometric analysis of the urine disclosed allylisopropylacetylurea (a fatty acid ureide sedative), diphenhydramine, chlorpheniramine and dihydrocodeine. The cause of death was considered to be drowning due to a drug overdose and cold exposure. The concentrations of diphenhydramine, free dihydrocodeine and total dihydrocodeine in the femoral venous blood (1.89, 3.27 and 3.30 microg/ml, respectively) were much higher than those in blood from the right cardiac chambers (0.294, 0.237 and 0.240 microg/ml, respectively). Urine concentrations of diphenhydramine, free dihydrocodeine and total dihydrocodeine were 22.6, 37.3 and 43.1 microg/ml, respectively. The stomach contained negligible amounts of diphenhydramine, free dihydrocodeine and total dihydrocodeine (0.029, 0.018 and 0.024 mg, respectively); concentrations of these drugs in the femoral muscle were 0.270, 0.246 and 0.314 microg/g, respectively. These results indicate that postmortem diffusion of diphenhydramine and dihydrocodeine from the bladder resulted in the elevated concentrations of these drugs in the femoral venous blood. Not only high urinary drug concentrations but also a large volume of urine in the bladder might accelerate the postmortem diffusion.

Adolescent↗

Sample pre-treatment for CO-oximetric determination of carboxyhaemoglobin in putrefied blood and cavity fluid.

Blood or blood-containing cavity fluid samples recovered from decomposed bodies in suspected carbon monoxide (CO) poisoning cases often have low total haemoglobin (tHb) levels or contain significant amounts of oily droplets, methaemoglobin (MetHb) and sulphaemoglobin (SHb), all conditions which render the validity of the measurements of carboxyhaemoglobin (HbCO) by commercially available CO-oximeters doubtful. Two sample pre-treatment methods, namely, centrifugal evaporation and addition of a commercially available standard haemoglobin solution containing a known level of HbCO, were developed to render such putrefied samples amenable to analysis by CO-oximeters. The methods developed were validated with samples of various degrees of putrefaction and various HbCO levels. The relative uncertainty of the determined HbCO levels as compared to their corresponding reference levels for the centrifugal evaporation method was approximately 5% and that for the standard addition method was <20%.

Body Fluids↗

Tissue distribution of nitrazepam and 7-aminonitrazepam in a case of nitrazepam intoxication.

We report a case of nitrazepam poisoning in which the distribution of nitrazepam and 7-aminonitrazepam was determined in body fluids and tissues. A 52-year-old woman was found dead in a shallow ditch (approximately 5 cm in depth), in the winter. Ambient temperature was 2-8 degrees C. The postmortem interval was estimated to be approximately 1 day and no putrefaction was observed. The cause of death was thought to be drowning due to nitrazepam overdose and cold exposure. Blood concentrations of nitrazepam and 7-aminonitrazepam were very site dependent (0.400-0.973 microg/ml and 0.418-1.82 microg/ml). In addition, the concentration of the same analytes in the bile were 4.08 and 1.67 microg/ml, respectively, and in the urine: 0.580 and 1.09 microg/ml, respectively. A high accumulation of both substances was observed in various types of brain tissue (2.17-6.22 microg/g and 2.49-5.11 microg/g). Only small amounts of nitrazepam and 7-aminonitrazepam were detected in the liver (0.059 and 0.113 microg/g, respectively). Large differences in the observed concentrations of nitrazepam and 7-aminonitrazepam among arterial and venous blood samples were thought to be mainly due to dilution of arterial blood by water entering the circulation through lungs at the time of death. Bacterial metabolism of nitrazepam may also have contributed to the observed differences.

Anti-Anxiety Agents↗

Into the decomposed body-forensic digital autopsy using multislice-computed tomography.

It is impossible to obtain a representative anatomical documentation of an entire body using classical X-ray methods, they subsume three-dimensional bodies into a two-dimensional level. We used the novel multislice-computed tomography (MSCT) technique in order to evaluate a case of homicide with putrefaction of the corpse before performing a classical forensic autopsy. This non-invasive method showed gaseous distension of the decomposing organs and tissues in detail as well as a complex fracture of the calvarium. MSCT also proved useful in screening for foreign matter in decomposing bodies, and full-body scanning took only a few minutes. In conclusion, we believe postmortem MSCT imaging is an excellent vizualisation tool with great potential for forensic documentation and evaluation of decomposed bodies.

Autopsy↗

Transcellular and extracellular impedances of the intact abdomen in putrefying rat cadavers.

Total (Zt) and extracellular (Ze) abdominal impedances of putrefying rat cadavers, stored at 18.0 +/- 0.7 degrees C, were measured at 50 kHz and at 1 kHz, respectively, during the first 7 days of death. Transcellular impedance was calculated from the measured values of Zt and Ze. Total impedance and Ze increased during the first day, and then decreased with increasing postmortem interval, while Zi remained unchanged during the first 3 days before increasing more than 4-fold between 3 and 7 days postmortem. These changes are consistent with net gain of electrolyte by the extracellular fluids and with degeneration of cell membranes which occur postmortem. It is concluded that, during putrefaction, Ze is a potentially more useful index of postmortem interval than Zt or Zi, and that measurement of total abdominal impedance at 50 kHz, rather than at 10 kHz, markedly improves the precision and accuracy of impedance measurement.

Abdomen↗

Origin of blood ethanol in decomposed bodies.

Problems related to blood contamination by other postmortem fluids in decomposed bodies (DB) make the interpretation of medicolegal blood alcohol levels (B EtOH) a very difficult task. So the aim of this paper is to show the utilization of vitreous humor (VH) as the biological fluid for an unequivocal determination of ethanol origin in DB for forensic purposes. Alcohol was determined in VH, blood (chest fluid-CF) and urine (Ur) collected from 27 DB in different states of putrefaction. A simple head-space gas-chromatographic method was used. In fifteen cases alcohol was found to be of endogenous production due to its absence in VH. In the twelve remainders, alcohol was detected in VH and CF in an atypical distribution. Examining the reliable scene and historical information together with the analytical data, ethanol origin in these cases was classified: endogenous production (3 cases), ingested (2 cases), both (2 cases), contaminated plus endogenous production (3 cases) and unable to determine (2 cases). According to the results obtained it was possible to conclude that alcohol analysis in VH is fundamental for determining the origin of ethanol detected in CF of DB.

Alcohol Drinking↗

The concentration of swine production. Effects on swine health, productivity, human health, and the environment.

The concern about environmental issues centering around CAFOs is appropriate. The veterinary profession can be an important force in meeting these challenges by broadening its scope of knowledge and practice into the broader environmental field. Although animal agriculture's contribution to environmental concerns is the focus of this article, it is only one of several sectors that contributes to environmental degradation. Crop production, as well as livestock production industries, contribute to pollution. Manufacturing industries, municipalities, private individuals, our consumptive lifestyles, and agriculture all contribute to the degradation of our environment. One must keep in mind the huge importance of our agricultural industry and not single it out to the detriment of its progress. We have an abundance of high-quality foods at the lowest cost to the individual of any industrialized nation. We export over 40 billion dollars in agricultural products yearly. Agriculture sustains our rural economies and provides opportunities for over 2 million private enterprises scattered across the country; however, there is a goal that we have a sustainable agriculture. A big part of that depends on development and enhancement of an agriculture that does not pollute, that sustains its farm operators and workers, and that does not make the area residents ill or degrade their quality of life; however, the current situation is not promising. Much remains to be learned about the actual acute and long-term health consequences of animal agricultural pollution. Many health concerns are speculative, even though based on sound facts. We know that many surface waters have excess N and P that leads to eutrophication and possibly enhanced growth of undesirable organisms such as Pfiesteria piscicida. We know that other animal pathogens, such as cryptosporidia, have caused large community outbreaks. There are other potential pathogens, such as Salmonella sp, for which we do not know the hazard. We know that our soils may become excessively laden with P, Cu, and Zn, which retard plant growth and create toxic conditions for grazing animals. There are concerns about air pollution. Odors have negative sensory and physical health consequences. H2S and dust may cause toxic effects on neighbors. NH3 vaporizing from manure sources may be carried with precipitation to cause eutrophication in lakes or altered ecosystems in natural areas. CH4 escaping from degrading manure contributes to greenhouse gases. Workers in confined livestock structures have high risk for a variety of chronic respiratory conditions. They also are at risk for acute poisoning from H2S in operations where liquid manure is stored in confined spaces. There have been numerous health complaints in recent years from community neighbors of large-scale livestock operations. One study showed adverse altered mood states, and another showed evidence of respiratory illness similar to what workers experience. Although it has not been possible to objectively measure conditions and know toxic levels of substances causing these illnesses, there are so-called extratoxic mechanisms, such as inherent aversion to putrefactive odors and exacerbation of preexisting conditions that lower the tolerance threshold. Environmental concerns regarding livestock production are not new. In the early and mid-1970s, there were many conferences and publications regarding odors and water contamination from livestock operations. Although most of what is known in this area has been known for 20 years, relatively little effective efforts have been made to correct the concerns. In fact, trends over this past decade have increased the concerns. This past decade has seen a tremendous acceleration in the concentration and consolidation of agriculture, capping a slow trend over the past 50 years toward larger, fewer, and more-specialized farms. This trend has gone against the old saying that "dilution is the solution to pollution.

Air Pollutants↗

Effects of anaerobic/aerobic incubation and storage temperature on preservation and deodorization of kitchen garbage.

To develop a garbage recycling system for the purpose of the production of lactic acid (LA) to use as raw material for producing biodegradable plastics, the preservation and deodorization of garbage during storage are very important. Anaerobic incubation (i.e., storage) was prove to be more suitable than aerobic incubation during the garbage storage in terms of concentration of LA and soluble sugar, pH value, viable bacteria counts and offensive odour substances. This difference is due to a fact that the growth of putrefactive bacteria such as coliforms and Clostridium spp. appeared to be inhibited by anaerobic fermentation during the storage, because the fermentation caused a drop of garbage pH and generated inhibitory substances, i.e., bacteriocins. Under anaerobic condition, LA concentration in the stored garbage was found to be higher in the order: 37 > 25 > 50 > 5 degrees C, and the concentration of sugar accumulated during the 50 degrees C-storage was the highest. Among the conditions employed, the optimum condition for the storage of kitchen garbage was anaerobic at 5 degrees C.

Bacteria, Aerobic↗

Criteria for judging whether postmortem blood drug concentrations can be used for toxicologic evaluation.

Postmortem blood drug concentrations tend to be regional-dependent. The aim of this study was to establish preliminary criteria, based on analytical findings of drugs in body fluids, for objectively judging whether drug concentrations determined in postmortem blood are usable for toxicologic evaluation. The study involved 11 autopsy cases in which no obvious putrefaction was observed. Twelve drugs were assayed; the cumulative frequency of detection was 16. The drug concentrations in cerebrospinal fluid and pericardial fluid were averaged, and the ratio between this average concentration and the concentration in femoral venous blood was determined. The mean ratio was close to 1, with a small standard deviation (0.94 +/- 0.20). From these results, the following criteria were drawn: 1) when the ratio of average cerebrospinal fluid/pericardial fluid drug concentration to blood drug concentration is within a range of 0.6-1.4, the postmortem blood drug concentration is usable for toxicologic evaluation; and 2) when the ratio is outside this range, the average cerebrospinal fluid/pericardial fluid concentration should be used as an alternative to drug concentration is postmortem blood.

Journal Article↗

Pericardial fluid as an alternative specimen to blood for postmortem toxicological analyses.

In actual forensic cases, we occasionally encounter victims with their blood being completely lost. In this study, pericardial fluid has been proposed as a specimen for toxicological analysis, and its utility has been evaluated. Fifteen autopsy cases with little putrefaction were selected. Fairly good correlations were observed between blood and pericardial fluid for all drugs, neutral and basic drugs and acidic drugs with regression equations of y=1.09x - 0.086 (r=0.989, n=21), y=0.969x - 0.072 (r=0.993, n=16) and y=1.01x + 0.355 (r=0.970, n=5), respectively. The correlations of drug concentrations between blood and cerebrospinal fluid/femoral muscle were not as good as those between blood and pericardial fluid. No correlations were observed between blood and urine/bile. The ratios of pesticide concentrations in each specimen to those in blood showed a large variation. Although our study was limited to a small number of cases, we have concluded that pericardial fluid is a good sample for quantitative confirmation of analyses performed on blood samples or a quantitative alternative to blood in exsanguinated victims. Cerebrospinal fluid, urine, bile and the skeletal muscle were found to be suitable only for qualitative analyses.

Journal Article↗

Rainfall and temperature effects on the decomposition rate of exposed neonatal remains.

There are scarce data describing seasonal effects on decomposition, and on the decomposition of neonatal remains. Piglet carcasses were exposed in a forest at Coranderrk Bushland (Victoria, Australia) each season for two years. The effects of season, year, temperature, and rainfall on mass loss rate and decay stage durations were investigated. There was a significant season and year interaction in weeks taken until the end of putrefaction: this was longest in winter, followed by autumn, spring and summer. Higher temperatures and rainfall increased both mass loss rates, and decomposition stage progression rates.

Animals↗

Purification and characterization of a protease from Pseudomonas aeruginosa grown in cutting oil.

The Pseudomonas aeruginosa san-ai strain was isolated from water-soluble cutting oil used for cooling and lubrication during industrial metal-working processes. This strain, which is grown in a high alkaline (pH 10) mixture of surfactants and mineral oil, produces an extracellular proteolytic enzyme. We have purified and characterized this 18 kDa protease. The P. aeruginosa san-ai protease functions optimally at pH 9.0 and 60 degrees C. Additionally, it is a Zn-containing metalloenzyme, and its monomeric structure contains at least one disulfide bond. Because the enzyme is stable in the presence of organic solvents, it is suitable for peptide synthesis. Furthermore, the P. aeruginosa san-ai protease could be used in an intelligent drug delivery system (DDS) designed for applications in the metal industry for prevention of putrefaction of cutting oil.

Journal Article↗

Ecology of anti-microbials produced by bacterial associates of Steinernema carpocapsae and Heterorhabditis bacteriophora.

Based on the ability of bacterial associates of entomopathogenic nematodes to produce antibiotic compounds on artificial media, it has been commonly accepted that Xenorhabdus sp. and Photorhabdus sp. inhibit a wide range of invading microorganisms in insects infected with Steinernema spp. or Heterorhabditis spp. Therefore, the question of whether antibiotic compounds produced by the primary form of bacterial symbionts associated mutualistically with S. carpocapsae and H. bacteriophora explain why insect carcasses do not putrefy but provide nutritional requirements for insect parasitic rhabditoid nematodes to complete their life-cycle was examined. Laboratory bioassays of anti-bacterial activity on nutrient agar and during parasitism in larvae of Galleria mellonella have confirmed earlier observations that in virto colonies of the primary form of X. nematophilus and P. luminescens produced agar-diffusible antibiotic compounds of a broad spectrum of anti-bacterial activity; their role in parasitism seems doubtful, however. This hypothesis is supported by a low antibiotic potency of a limited spectrum of anti-bacterial activity throughout the life-cycle of the parasites, principally in Galleria infected with S. carpocapsae. Since the lack of putrefaction cannot be explained simply by antibiotic inhibition of contaminating bacterial microflora, other competition mechanisms must be operating in parasitized insects. I postulated that a rapid and massive colonization of the insect body by nematophilic bacteria creates unfavorable conditions for the growth and multiplication of bacterial (proteolytic) contaminators making the insect carcass decay-resistant. In the case of H. bacteriophora, low antibiotic activity at an early stage of parasitism could support the colonization by P. luminescens of the host.

Animals↗

Halitosis and Helicobacter pylori: a possible relationship.

With the aim of investigating a possible relationship between "objective" halitosis (established by sulfide levels in the breath) and Helicobacter pylori, we performed a study in 58 dyspeptic patients reported to suffer from "bad breath." Furthermore, we evaluated the effects on halitosis of eradication therapy (only for H. pylori-positive patients) and chlorhexidine antiseptic mouth rinses (in all patients). Sulfide compound assay indicated objective halitosis in 52/58 patients, 30 of whom were positive and 22 negative for H. pylori. In 19/30 eradication by double therapy provoked a decrease to below the cutoff value of sulfide levels in 15. In the other 11 of the 30 subjects, in whom H. pylori positivity persisted, halitosis parameters did not change. Chlorexidine reduced sulfides to below the cutoff value in 16/22 H. pylori-negative patients, but did not provoke any change in the 11 unsuccessfully treated H. pylori-positive subjects. In these, objective halitosis disappeared only after a successful eradication by triple therapy (9/11). Our results show a possible association between halitosis and H. pylori since bacterial eradication may resolve the symptom. Antiseptic mouthwashes may be effective only in absence of H. pylori, when halitosis may be due to oral putrefactive microbial activity. In a small number of subjects the cause and treatment of halitosis need to be clarified.

Adolescent↗