Chlorpromazine-induced alterations in hypothalamic amine metabolism and stress responses in severe cold

To investigate the effects on the central nervous system of severe cold stress with and without chlorpromazine, guinea pigs were treated with chlorpromazine or 0.9% NaCl and exposed to -20 degrees C or +23 degrees C for 1 h. Hypothalamic noradrenaline (NA), dopamine (DA), 5-hydroxy-tryptamine (5-HT), 3-methoxy-4-hydroxyphenyl ethylene glycol (MHPG), homovanillinic acid (HVA) and 5-hydroxy-indoleacetic acid (5-HIAA) were determined by high-performance liquid chromatography. Serum, urinary and vitreous fluid catecholamines, muscle and liver glycogen, and blood glucose were also measured. Chlorpromazine caused distinct hypothermia at -20 degrees C and slight hypothermia at +23 degrees C. The rise in hypothalamic MHPG, 5-HIAA and MHPG/NA and in 5-HIAA/5-HT ratios in the cold indicate increased noradrenergic and serotonergic activity. The latter was inhibited by chlorpromazine and a drug-induced inhibition of noradrenergic neurons could not be ruled out. Chlorpromazine increased the turnover of DA at room temperature and the same tendency was seen in the cold. The hypothermic animals had low serum catecholamines, indicating diminished sympathetic activity. The chlorpromazine-treated cold-exposed animals did not react to the environmental stress by sympathetic activation, as urinary NA and adrenaline were not elevated, but DA was excreted by all the drug-treated animals. Vitreous fluid NA and DA were elevated as an indicator of cold stress, and no drug effect was seen in this fluid.     

Vital reactions to frostbite of the ear and paw skin in guinea pigs exposed to the cold

Vital reactions to frostbite in the paw and ear skin of guinea pigs were studied in order to find an animal model for frostbite in cases of accidental hypothermia. One group of animals was rendered hypothermic (rectal temperature, 30 degrees C) by exposure to an ambient temperature of -20 degrees C, and samples were taken from the frozen skin. A second group was rendered hypothermic and rewarmed in warm air at 45 degrees C, and samples were taken from the thawed skin. The only vital reaction in the first group (freezing time, 4-5 h) was mild initial inflammation, which was expressed in granulocyte adhesion to the vessel wall and the migration of a few cells into the dermis. The inflammatory reaction was more distinct in the second group (freezing and thawing together 5-7 h), with a large number of granulocytes being present in the dermis. Oedema and hyperaemia were also present in the frostbitten tissue after thawing, but no signs of necrosis developed. The alkaline-phosphatase reaction demonstrated the presence of granulocytes more clearly than H & E or Masson trichrome staining. Vital reactions were more advanced in the ear skin. It is concluded that vital reactions are very scarce in cases of frostbite, even after several hours' exposure, unless the tissue is allowed to thaw.     

Complement component C3a or C3a desArg as a new marker for estimation of local vital reactions in incised skin wounds.

The anaphylatoxin C3a or its desArg form (C3a/desArg) generated during complement activation could be detected in the vicinity of incised skin wounds of guinea pigs using immunoblotting methods. The C3a/desArg peptides were detectable immediately after injury in local sites up to 3 mm from the wound edge. In subsequent determinations of up to at least 3-day-old antemortem wounds, the maximum concentration of these peptides was largely localized up to 6 mm from the wound edge at 2 h after injury. In postmortem wounds, however, these peptides were undetectable. When they were released in antemortem wounded tissues they could be detected up to 1 day at 22 degrees C after death. These results suggest that the detection of C3a/desArg in wounds using immunoblotting methods can be useful for distinguishing ante- from postmortem wounds.     

Time of death of victims found in cold water environment

Limited data is available on the application of post-mortem temperature methods to non-standard conditions, especially in problematic real life cases in which the body of the victim is found in cold water environment. Here we present our experience on two cases with known post-mortem times. A 14-year-old girl (rectal temperature 15.5 degrees C) was found assaulted and drowned after a rainy cold night (+5 degrees C) in wet clothing (four layers) at the bottom of a shallow ditch, lying in non-flowing water. The post-mortem time turned out to be 15-16 h. Four days later, at the same time in the morning, after a cold (+/- 0 degrees C) night, a young man (rectal temperature 10.8 degrees C) was found drowned in a shallow cold drain (+4 degrees C) wearing similar clothing (four layers) and being exposed to almost similar environmental and weather conditions, except of flow (7.7 l/s or 0.3 m/s) in the drain. The post-mortem time was deduced to be 10-12 h. We tested the applicability of five practical methods to estimate time of death. Henssge's temperature-time of death nomogram method with correction factors was the most versatile and gave also most accurate results, although there is limited data on choosing of correction factors. In the first case, the right correction factor was close to 1.0 (recommended 1.1-1.2), suggesting that wet clothing acted like dry clothing in slowing down body cooling. In the second case, the right correction factor was between 0.3 and 0.5, similar to the recommended 0.35 for naked bodies in flowing water.     

Skeletal muscle activity of alcohol-treated rabbits in immersion hypothermia: adenine nucleotide concentrations and phosphorylation state

The effects of alcohol injection (0.5 g . kg-1 i.v.) on the core cooling and rewarming rates, concentration of the adenine nucleotides, and the phosphorylation state of the adenylate system (ATP/ADP X P) were studied in the skeletal muscle of anesthetized rabbits immersed in ice-cold water. NaCl-injected rabbits immersed in ice-cold water were used as cold controls, alcohol-treated animals at room temperature (20 degrees C) as alcohol warm controls, and NaCl-injected animals at room temperature as anesthesia controls, respectively. The fall of core temperature to 32 degrees C in the alcohol-treated rabbits and the cold controls took about 40 min. During this time the temperature of the alcohol warm and anesthesia controls fell by about 1 degree C. No difference in the rewarming rate was observed between the alcohol-treated and cold control rabbits. Serum glucose concentration was elevated in the cold controls (from 5.9 to 8.3 mmol/l) but not in the alcohol-treated rabbits. Cold exposure reduced the phosphorylation state in the skeletal muscle of the alcohol-treated rabbits by 32% (P less than 0.05), but the decrease (6%) was not significant in the cold controls. After rewarming the phosphorylation state decreased in the above groups by 71% and 15%, respectively, as compared with the initial values. No significant changes in the phosphorylation state were found in the warm control animals. The redox state of the cytosol in the skeletal muscle or liver did not change, nor was there any change observed in the arterial pO2 or pCO2 concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)     

豚鼠过敏性休克肺组织中类胰蛋白酶和胃促胰酶的表达

目的观察豚鼠过敏性休克死亡肺组织中类胰蛋白酶和胃促胰酶的表达情况,试图为过敏性休克死亡提供客观的诊断依据。方法健康豚鼠24只,随机均分为实验组和对照组,每组再分死亡即时组、冷藏48h组和冷冻7d组,每组4只。实验组将0.5mL人混合血清用生理盐水1∶10稀释,注射于豚鼠后掌皮内,致敏后3周以人混合血清1mL注入心腔诱发过敏性休克致死;对照组采用生理盐水代替混合血清。提取豚鼠心血及肺组织,应用免疫组化染色和图像分析技术观察类胰蛋白酶和胃促胰酶的表达情况。结果对照组豚鼠肺组织中类胰蛋白酶和胃促胰酶阳性细胞数量较少,分布在小血管和小气管周围。实验组肺组织中类胰蛋白酶和胃促胰酶阳性细胞明显增多,多数细胞形态不规则,阳性染色颗粒脱出肥大细胞并弥散到组织间隙。冷藏48h和冷冻7d的条件下对这两种酶的表达无明显影响。结论过敏性休克致死豚鼠肺组织中类胰蛋白酶和胃促胰酶的表达明显增强,在冷藏48h和冷冻7d内的条件下,可作为过敏性休克死亡的一项诊断依据。     

过敏性休克急死豚鼠血清和肺组织内IgE的变化及法医学意义

目的研究过敏性休克急死豚鼠死后在4℃冷藏条件下血清IgE含量和肺组织内IgE的免疫表达随死后不同时间的变化,探讨过敏性休克急死的法医学鉴定的客观诊断指标。方法采用多人混合血清注射建立过敏性休克急死的动物模型。豚鼠40只,随机分为对照组和实验组,实验组又分为死后0、12、24和48h组,每组8只。采用酶联免疫吸附试验测定豚鼠血清IgE含量,采用免疫组化ABC法对豚鼠肺组织IgE进行免疫组化染色。结果实验组与对照组豚鼠血清IgE含量相比较有显著性差异(P<0.001);死后0、12、24和48组血清IgE含量相比较无显著性差异(P>0.05)。实验组豚鼠肺组织IgE有阳性表达,对照组豚鼠无IgE表达。结论过敏性休克急死豚鼠血清IgE含量显著升高;在4℃下冷藏,死后48h内血清IgE含量和肺组织中IgE无明显变化。本结果可为过敏性休克急死的法医学鉴定提供客观的诊断依据。     

Sympatho-adrenal activity in acute cold stress. The mechanism of sudden death following water immersion]

In addition to currently known mechanisms of sudden death following water immersion, predominantly vagal cardio-depressive reflexes are discussed. The pronounced circulatory centralization in diving animals as well as following exposure to cold water indicates additional sympathetic activity. In cold water baths of 15 degrees C, our own measurements indicate an increase in plasma catecholamine levels by more than 300%. This may lead to cardiac arrhythmias by the following mechanism: Cold water essentially induces sinus bradycardia. Brady- and tachyarrhythmias may supervene as secondary complications. Sinusbradycardia may be enhanced by sympathetic hypertonus. Furthermore, ectopic dysrhythmias are liable to be induced by the strictly sympathetic innervation of the ventricle. Myocardial ischemia following a rise in peripheral blood pressure constitutes another arrhythmogenic factor. Some of these reactions are enhanced by alcohol intoxication.     

Anaphylactic death: the effect of aminoguanidine and heparin on histamine and stress hormones in guinea pigs

The modifying effect of aminoguanidine (a histaminase inhibitor) and heparin (a histaminase liberator) on anaphylactic shock in guinea pigs was studied using ovalbumin as an antigen and trigger. The animals died of the shock, the time to death remaining unaltered by the drugs. Serum histamine and cortisol values were high after shock, but were reduced by heparin. Both noradrenaline and adrenaline in plasma were also elevated after shock, the final concentration of the latter being lowered by heparin. The lungs were dilated, indicating bronchoconstriction. The results confirm the role of histamine in anaphylactic shock and its potential value for the diagnosis in this kind of rapid death, in which morphological signs are scarce or lacking. Its diagnostic value still requires confirmation, however, which only autopsy studies can supply. It also appears that pretreatment of the animals with heparin affected the blood cortisol and catecholamines, which are involved in the shock mechanism as countermeasures, although aminoguanidine did not have any effect.     

Characteristics of fibrin formation in stab wounds of the skin, determined by scanning electron microscopy

In an investigation on fibrin formation in skin cut wounds on guinea pigs using scanning electron microscopy, it was established that immediately after the infliction of cuts on live animals, a fine, netlike fiber structure of fibrin forms that gradually covers the entire wounded surface. In the period early after the cuts blood-forming elements attach to the fibrin net through numerous fine fibrin fibers, which incorporate a great number of thrombocytes. Postmortem investigations of these wounds showed that the fibrin net formed preserves its structure. It was also established that a fibrin net forms over skin wounds that were inflicted at different periods after death. In the wounds in the early periods after death, the fibrin net formed was very similar to the one established on wounds inflicted on living animals. Our investigation describes some peculiar characteristics of the fibrin net formed in living animals; there is a fine fiberlike net and the quantity is definitely larger. These characteristics permit the establishment of whether the wounds occurred before death and how much time has lapsed since then.     

A new biochemical method for estimation of postmortem time.

Hypoxanthine (Hx) is formed by hypoxic degradation of adenosine monophosphate (AMP) and might be elevated due to antemortem hypoxia. However, it also increases after cessation of the life processes. Until now measurements of potassium in corpus vitreous humor have been used by forensic pathologists to determine postmortem time. In this study the influence of postmortem time and temperature on vitreous humor Hx and potassium levels were compared. Repeated sampling of vitreous humor was performed in 87 subjects with known time of death and diagnosis. The bodies were kept at either 5 degrees C, 10 degrees C, 15 degrees C or 23 degrees C. Hx was measured by means of HPLC and potassium by flame photometry. In 19 subjects from whom samples were obtained within 1.5 h after death, the normal level of Hx could be estimated to be 7.6 mumol/l and that of potassium to be 5.8 mmol/l. The spread of the potassium levels measured shortly after death was much greater than for the corresponding Hx levels. In the four temperature groups the Hx level increased 4.2, 5.1, 6.2 and 8.8 mumol/l per h, respectively, whereas the corresponding figures for potassium were 0.17, 0.20, 0.25 and 0.30 mmol/l per h. The vitreous humor concentration of both Hx and potassium increases fairly linearly after death. The slopes are steeper with increasing temperature. Since the scatter of the levels is greater for potassium than for Hx, the latter parameter seems to be better suited for the determination of time of death in cases without antemortem hypoxia, especially during the first 24 h.     

甲基苯丙胺及其代谢产物在急性中毒豚鼠体内的分布

目的研究甲基苯丙胺 (MAP)及其代谢产物苯丙胺 (AP)在急性中毒豚鼠体内的含量分布。方法应用GC/NPD技术 ,以 4 苯基丁胺 (4 PBA)为内标 ,样品经水解后碱化或直接碱化至pH >11,环已烷混旋提取 ,三氟乙酸酐 (TFA)微波衍生化 ,测定MAP急性中毒豚鼠体液和组织中MAP及AP的含量。结果急性中毒死亡豚鼠体内各器官和体液中MAP及AP含量最高为肺 ;其次为肝、脑、肾、脾、肠、心、血 ;再次为胃、胆汁 ;最少是尿。结论MAP在动物体内代谢迅速 ,组织或体液中MAP和AP浓度的比值与豚鼠给药后存活时间有关     

Changes in the amount of prostaglandin F2 alpha in incision wounds of the skin with different time intervals after injury

A radioimmunological method is presented for the determination of the quantity of prostaglandin F2 alpha in cut wounds on the skin of guinea pigs. Rapidly increasing quantities of prostaglandin F2 alpha can be found in skin cuts, and the level reaches 71 ng/g within the 1st hour after the injury. In the postmortem period, the quantities calculated in the cuts while the animals were still alive gradually decreased and reached a value of 17 ng/g in the 6th h after death. In postmortem cuts, inflicted in the 8th h after death, the prostaglandin was 14-18 ng/g. In later postmortem cuts the quantity was about 9-10 ng/g. Establishing the dynamics of the quantitative changes permits investigation of the prostaglandin to be used to certify whether the victim was alive or not, as well as when the skin damage was inflicted.     

Postmortem stability and interpretation of beta 2-agonist concentrations.

This paper describes a series of stability and redistribution studies aimed at understanding the presence and significance of beta 2-agonists in asthma deaths. Salbutamol and terbutaline were shown to be stable in postmortem blood at 23 degrees C for 1 week, 4 degrees C for 6 months and -20 degrees C for 1 to 2 years. However, fenoterol was shown to degrade at 23 degrees C (83% loss), 4 degrees C (93% loss) and -20 degrees C (66% loss) over the same time. Salbutamol concentrations detected in blood taken at the time of body admission to the mortuary were not significantly different from the concentrations detected in blood taken from the same cases at the time of autopsy (45 h later). This suggests that significant postmortem redistribution of salbutamol is unlikely to occur during this period. Postmortem blood concentrations of at least salbutamol are likely to reflect the concentration of these drugs in the body at the time of death.     

Identification and significance of delta-aminovaleric acid in putrefaction materials (author's transl)]

During former putrefaction experiments regularly a proteogenic substance has been found which by means of modern analytical methods now was identified as delta-aminovaleric acid (DAVA). DAVA seems to appear in guinea pig as well as human organs and some body fluids under experimental conditions never before the 3rd (20 degrees C) to 5th day (10 degrees C). It is characterized by statistically significant increases until the end of the 2nd (20 degrees C) to 5th week (10 degrees C) and relatively stable values thereafter. Considering storage temperature measurement of DAVA concentration can be of relevance for the estimation of the time of death in cases of putrescent corpses.     

The Effects of Continuous Application of the TASER X26 Waveform on Sus scrofa,

This study investigated and evaluated the safety margins of the continuous long duration (up to 30 min) effect of the TASER X26 waveform, using a Sus scrofa model. Long duration continuous stimulus has not been evaluated on humans or human surrogates prior to this study. Swine were used as models due to similarities with humans in their skin and cardiovascular systems. Very long duration was used to determine both exposure dose and possible adverse physiological effects of dose. The trial began with an application of 10 min, and subsequent animals received increasing exposure time up to a survived maximum duration of 30 min. At the onset of this work, it was hypothesized that there would be a time limit after which most animals would not survive consistent with increased dose response. However, this hypothesis was not supported by the experimental results. All animals (10 of 10) survived up to 3 min. Seven of the 10 animals survived up to a 10‐min exposure and 3 of 5 animals with a 30‐min target exposure survived the full exposure. Surviving animals were recovered and observed for 24 h, with no postrecovery deaths. This suggests that swine (based on physiology) will not experience a fatal event when exposed to the TASER X26 for a continuous 3 min. Conclusions regarding longer duration (10–30 min) are not as certain due to the small sample sizes at these time intervals.     

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.     

Creatine phosphokinase in serum and cerebrospinal fluid, and microscopic findings in brain and heart in hypothermic rabbits

Signs of hypothermia injury were studied in rabbits cooled to a core temperature of 30 degrees C by immersion in ice water and thereafter rewarmed to 35 degrees C. Anaesthetized control rabbits were kept normothermic (37 degrees C) for a corresponding time (4 h). Creatine phosphokinase (CPK) activity increased 24 h after hypothermia to 20-fold in serum. In cerebrospinal fluid the activity was already significantly (5-fold) increased after hypothermia and was still as high at 24 h. Smaller increase was also found in the control normothermic rabbits both in serum (10-fold) and cerebrospinal fluid (2-fold). The values had returned to the initial level after 1 week. Small haemorrhages were observed in the brain at 24 h and slight scarring was seen in the myocardium of some rabbits which had lived 4 weeks following hypothermia. The results indicate that CPK can be a useful marker in the diagnostics of hypothermia death, especially in cerebrospinal fluid, which is less affected than blood by autolysis.     

过敏性休克豚鼠血浆咽喉肺组织中P物质的观测

目的探讨过敏性休克法医学鉴定的诊断指标。方法制备豚鼠过敏性休克的动物模型,应用放射免疫法测定其血浆中P物质浓度;免疫组化SABC法和BI-2000图像分析系统对咽喉和肺组织的P物质免疫反应进行染色观测,计算阳性指数(PI)。结果与对照组含量(87．70pg±7．60pg／μl)相比,过敏性休克豚鼠血浆中P物质含量 (131．01pg±18．93pg／μl)增加,其差异具有极显著意义(P<0．01);呼吸道内P物质免疫阳性反应增强,实验组咽喉和肺组织PI值分别为63．59±14．51和55．98±14．8,对照组咽喉和肺组织PI值分别为33．32±8．04和20．51±6．76, 两组差异具有极显著意义(P<0．01)。结论 P物质在过敏性休克豚鼠血浆浓度增加和呼吸系统组织中免疫阳性反应增强,可能有助于过敏性休克的法医病理学诊断。     

大鼠死后心血吗啡浓度变化的HPLC检测

采用高效液相邑谱分析技术（HPLC）检测治疗量及中毒量吗啡肌注大鼠死后心血中吗啡浓度变化。结果表明，以治疗量吗啡肌往大鼠，在死后96h内，心血中吗啡浓度随死后时间增加而显著升高（P＜0．01），吗啡浓度水平与死后时间里显著正相关；以中毒量吗啡肌注大鼠，在死后12h内，心血吗啡浓度无明显变化；死后24h、48h及96h，随死后时间延长，心血中吗啡浓度逐渐升高（P＞0．01），其递增强度不如治疗量吗啡组大鼠的明显.本研究证实，死后尸体心血吗啡浓度明显受生前剂量的影响，且在死后96h内，随死后时间增加．心血中吗啡浓度少数不断增高。