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PubMed2006

Immunohistochemical distribution of cyclic nucleotide phosphodiesterase (PDE) isoenzymes in the human vagina: a potential forensic value?

OBJECTIVES: Phosphodiesterase (PDE) isoenzymes are key proteins involved in the maintenance of the normal function of various tissues of the human body including those of the male and female urogenital tract. More recently, PDEs and their main substrates, cyclic GMP and cyclic AMP, have also been assumed to play a crucial role in the control of the human vagina. In order to elucidate the potential significance of phosphodiesterases as marker proteins in female genital organs, it was the aim of the present study to evaluate by means of immunohistochemistry the distribution of cGMP- and cAMP-PDE isoenzymes in specimens of the human vagina. METHODS: Conventional immunohistochemical techniques (double antibody technique, laser fluorescence microscopy) were applied to sections of the human vaginal wall in order to evaluate the presence of the PDE isoenzymes 1, 2, 3, 4, 5 and 10. RESULTS: Immunoreactivities (IR) specific for PDE1 (cAMP/cGMP-PDE, Ca(2+)/Calmodulin-dependent), PDE2 (cAMP-PDE, cGMP-dependent) and PDE5 (cGMP-PDE) were exclusively registered in the smooth musculature of vaginal arterial vessels, whereas no signals were detected in non-vascular tissue. IR indicating the expression of the cAMP-degrading PDE4 was mainly observed in the vaginal epithelium. Vaginal epithelial cells also presented immunosignals specific for PDE3 (cAMP-PDE, inhibited by cGMP) and PDE10 (dual substrate PDE), nevertheless, these stainings were less abundant than those related to the PDE4. IR for PDE10 was also registered in inflammatory cells located in the subepithelial region of the vaginal wall. CONCLUSION: Our study revealed the presence of IR specific for PDE1, PDE2, PDE4, PDE5 and PDE10 in sections of the human vagina and demonstrated that these enzymes are not evenly distributed in the tissue. Especially, the prominent expression of the cyclic AMP-PDE4A in the vaginal epithelium may give hint to a potential significance of this isoenzyme as a forensic marker protein. The findings give a rationale to investigate further as to whether the immunohistochemical detection of PDE4 may represent a new forensic tool in order to identify human vaginal epithelial cells.

Adolescent↗
PubMed2006

Survival following accidental ligature strangulation: a case report.

Survival following accidental ligature strangulation is quite rare. The present case involves an adult male strangulated by a soft cotton cloth entangled in the rotor of a machine. Unilateral neck compression allowed survival of the victim. The victim escaped with minimal injuries which were limited to contusion of the neck and edema of the vocal cords and inter arytenoid region.

Accidents, Occupational↗
PubMed2005

Combined pH/organic solvent gradient HPLC in analysis of forensic material.

A combined pH/organic solvent linear gradient mode in high performance liquid chromatography (HPLC) is presented as a new approach to determination of low concentrations of ionogenic analytes in biological material. The approach consists in simultaneous development of linear gradients of pH and organic modifier in the mobile phase. Advantages of the method are illustrated in postmortem analysis of opipramol in material from suicide victims. Very narrow peaks without tailing were obtained and several times lower limits of analyte quantitation were achieved using ultraviolet detection as compared to a standard isocratic method. The double gradient HPLC method seems to be especially valuable in case of ionogenic analytes dispersed in complex biological matrices. That is due to a high selectivity of the double gradient method and the lack of peak tailing, which is commonly observed for basic analytes chromatographed at isocratic conditions.

Chromatography, High Pressure Liquid↗
PubMed2005

Cerebral concentrations of myo-inositol in rats with induced brain death.

In anaesthetized, mechanically ventilated rats with brain death induced by epidural balloon inflation, we assessed the levels of cerebral myo-inositol in three brain areas 4 h after brain death had been confirmed. The levels were measured using HPLC, along with the water content. Myo-inositol levels were significantly increased in the cerebellum (P<0.05) and decreased in the brainstem (P<0.001), compared to findings in controls. Such changes can serve as hallmarks of brain death.

Animals↗
PubMed2005

Electrolyte analysis in pleural effusion as an indicator of the drowning medium.

In medico-legal autopsies for drowned bodies, the location of drowning needs to be determined. To investigate the usefulness of electrolyte analysis in pleural effusion as an indicator of the location where the deceased has drowned, we determined the concentrations of electrolytes in the pleural effusion of rats drowned in four kinds of water. The concentrations of sodium and chloride ions in the pleural effusion of rats that drowned in seawater were significantly greater than those of rats that drowned in freshwater at both 1 day and 3 days after drowning. The concentration of potassium ions in pleural effusion 1 day after drowning showed no difference between each group, although it then increased from 1 to 3 days after seawater drowning, whereas it decreased from 1 to 3 days after freshwater drowning. The concentration of total protein in pleural effusion increased from 1 to 3 days after drowning, however, there was no significant difference in the concentration of total protein in pleural effusion between each group at either 1 day or 3 days after drowning. These results suggest that analysis of electrolytes in pleural effusion may be useful for determining whether drowning has occurred in seawater or in freshwater.

Animals↗
PubMed2005

HPLC analysis of anti-malaria agent, chloroquine in blood and tissue from forensic autopsy cases in Tanzania.

HPLC analysis of anti-malaria agent, chloroquine (CQ) in blood and tissues with a simple HCl back extraction method was applied to three forensic autopsy cases in Dar es Salaam, Tanzania. CQ concentrations in femoral vein blood were 8.5, 48.4 and 43.8 microg/ml in three cases, respectively, which were high enough to attribute the cause of deaths to an acute CQ poisoning. There were great site dependent variations in blood CQ levels. The right heart blood samples were very high, which may be explained by incomplete distribution of the drug before death or postmortem diffusion from liver and its surrounding blood, as high CQ levels were remarkable in the liver. Suicidal and accidental CQ poisonings are very common and CQ is a very important chemical in the field of forensic toxicology in Tanzania.

Adult↗
PubMed2004

Possible factors contributing to the postmortem lung weight in fire fatalities.

The aim of the present study was to examine the factors that contribute to the postmortem lung weight in acute fire fatalities (n=149) including those with lower (<60%) and higher (>60%) blood carboxyhemoglobin (COHb) levels (n=94 and 55, respectively). The control groups consisted of acute myocardial infarction/ischemia (AMI, n=99) and mechanical asphyxiation (n=85). For all cases (n=333), the lung and heart weights were independent of the postmortem time (4.5-72 h) and charring of the body. The combined weight of both lobes of the lung showed a significant gender difference (males>females, P<0.001), negative regression with respect to age (R=0.167, P<0.01) and positive regression with respect to heart weight (R=0.316, P<0.001). The gender difference was also significant for each cause of death even after being corrected using body height measurements. When the lung-heart weight ratio was estimated to diminish the influence of possible cardiogenic factors, the gender difference was insignificant for each cause of death among the non-elderly (<60 years of age). However, for elderly fire fatalities (>60 years of age), there was a gender difference (males>females) for the lower COHb group (P<0.05) and higher COHb group (P<0.001). A significant age-dependent difference (non-elderly>elderly) in the lung-heart weight ratio was observed for fire fatalities with a lower COHb and AMI among males and for fire fatalities with a higher COHb among females. Such gender- and/or age-dependent influences were not significant for fatal mechanical asphyxiation. These findings suggest that a person's heart weight may be a possible contributory factor to an increase in the lung weight in acute death, and that gender- and/or age-dependent susceptibilities may be additional factors that contribute to fire fatalities and AMI. In addition, elderly females appear to be most susceptible among fire casualties, and extreme cardiomegaly may also be a potential fatal risk factor.

Adolescent↗
PubMed2005

Ubiquitin-immunoreactive structures in the midbrain of methamphetamine abusers.

Ubiquitin (Ub) is involved in neurodegeneration and various stress responses in the brain. The present study investigated the Ub-immunoreactive structures in the midbrain of methamphetamine (MA) abusers as a marker of drug-induced neurodegeneration. Medico-legal autopsy cases were examined: fatal MA intoxication (n=14), other fatalities of MA abusers (n=23) including those due to injuries, asphyxiation, drowning, fire and natural diseases, and control groups (n=260). In the motor nervous systems, MA abusers showed a mild increase in the diffuse-type nuclear Ub-positivity in the pigmented neurons of the substantia nigra, depending on the blood MA level and irrespectively of the immediate causes of death. The intranuclear inclusion-type Ub-positivity of the nigral neurons and the granular 'dot-like' Ub-immunoreactivity area in the crus cerebri (cortico-spinal tracts) were usually low in MA abusers, and any increases were related to the immediate causes of death and the age of subjects. Acute MA fatality showed a higher neuronal Ub-positivity in the midbrain periaqueductal gray matter (PGM), which is involved in processing pain, fear and anxiety, and regulation of respiration and circulation. These findings suggest dysfunction of the nigral dopaminergic neurons and PGM neurons in the midbrain in MA abuse, which may account for the clinical symptoms.

Adult↗
PubMed2005

Quantitative analysis of ubiquitin-immunoreactivity in the midbrain periaqueductal gray matter with regard to the causes of death in forensic autopsy.

The aim of the present study was to examine Ub-immunoreactivity in the midbrain periaqueductal gray matter (PGM), which is involved in pain processing and modulation, in forensic autopsy cases (n=273) in relation to the causes of death: acute deaths from blunt injuries (n=75), sharp weapon injuries (n=36), fatal asphyxiation (n=22), drownings (n=16: freshwater, n=9; saltwater, n=7), fire fatalities (n=64), poisoning (n=12), hyperthermia (n=5), hypothermia (n=5), delayed deaths from blunt head injury (n=8), acute cardiac deaths (n=24), and acute cerebrovascular strokes (n=6). The Ub-immunoreactivity was clearly observed in the nuclei of the PGM neurons, showing no postmortem interference or age-dependency. A higher value was observed in blunt injuries, fire fatalities and also in saltwater drowning, hyperthermia and delayed head injury deaths. These findings suggest a complicated mechanism for the ubiquitination of PGM neurons, to which multiple factors including the intensity and duration of pains possibly under alert consciousness, traumatic and metabolic neurodegeneration may contribute.

Adolescent↗
PubMed2005

Ubiquitin immunoreactivity in the midbrain as a marker of stress to motor nervous systems in fatal injury.

Previous studies showed an increase in the ubiquitin (Ub)-immunoreactive structures in the midbrain in acute deaths from asphyxiation and in fires in adult subjects. The present study examined the Ub-immunoreactivity in the midbrain as a marker of stress to motor nervous systems in fatal injury cases (over 35 years of age, n=140: blunt injuries, n=82; sharp injuries, n=58), compared with that in control groups (n=61) including death by strangulation, acute cardiac and cerebrovascular diseases. The intranuclear Ub-immunopositive inclusion of the pigmented dopaminergic neurons of the substantia nigra (inclusion Ub-index) and the granular 'dot-like' Ub-immunoreactivity area ('dot-like' Ub-area) in the crus cerebri were analyzed. In blunt injuries, those markers were high in abdomen and back injuries and low in head and chest injuries. The inclusion Ub-index was higher in the crush/pressure injury group than in those with injuries due to impact traffic accidents and falls. 'Dot-like' Ub-area was also low in falls. In sharp injury cases, cardiac injury with hemopericardium showed a higher inclusion Ub-index. These findings suggested that the stress to the motor nervous system may be very intense in crush/pressure injury and hemopericardium than in impact injury and fatal hemorrhages, respectively, and in abdomen and back injuries than in head and chest injuries.

Adult↗
PubMed2005

The relationship of a high level of serum beta-hydroxybutyrate to cause of death.

To examine the state of ketoacidosis in relation to the cause of death, three kinds of ketone bodies (acetoacetate, beta-hydroxybutyrate and acetone) were measured in postmortem serum. Of 100 autopsy cases, 22 had ketone body increasing pathophysiological conditions, overlapped in some cases, namely a poorly-nourished state (10 cases), alcoholic fatty liver damage (10), diabetes (5) and infectious disease (5). Of the 3, 11, 7 and 15 cases in which the beta-hydroxybutyrate concentration was greater than 10,000, 1000-10,000, 500-1000 and 200-500 micromol/l, 3 (100%), 8 (73%), 3 (43%) and 5 (33%), respectively, had one or more pathophysiological conditions that usually produce ketone bodies. Of the 64 cases in which the beta-hydroxybutyrate concentrations were less than 200 micromol/l, only 3 (5%) had some of these conditions. Cases showing high levels of ketone bodies tended to have pathophysiological states that can produce them, although the level of beta-hydroxybutyrate and these states did not show parallel relationships. When autopsy findings fail to explain the cause of death, a diagnosis as death caused by ketoacidosis would be reasonable if the serum beta-hydroxybutyrate level is over 1000 micromol/l and the body has pathophysiological conditions that tend to increase ketone bodies.

3-Hydroxybutyric Acid↗
PubMed2005

Effects of perimortem physical factors associated with death on exogenous ethanol concentrations in cardiac blood.

The purpose of this study was to determine whether perimortem physical factors associated with death affect exogenous ethanol concentrations in cardiac blood. Forty-one autopsies of alcohol-intoxicated decedents with no or little putrefaction were involved. Postmortem intervals ranged from 6 to 96 h at the time of autopsy. Our 41 cases consisted of 17 fire victims, 8 drowned persons, 3 blood-loss patients who underwent unsuccessful cardiopulmonary resuscitation (CPR) for 30-50 min, 3 blood-loss persons without CPR, three non-blood-loss patients who underwent unsuccessful CPR for 5-60 min, and seven people who died of other causes. Ethanol concentrations in right cardiac chamber blood ranged from 0.12 to 4.40 mg/g (mean 1.39 mg/g) in all cases. An excellent correlation was observed between ethanol concentrations in right cardiac chamber blood and femoral venous blood (y=1.01x-0.087, n=38, r2=0.990). The ratios of blood ethanol concentrations in the left to right cardiac chambers were significantly lower in blood-loss decedents with CPR (0.71+/-0.13) than other decedents (0.92+/-0.07-1.06+/-0.06). Two factors, blood-loss and prolonged chest compression under ventilation, might have caused a decrease in blood ethanol concentration in the left cardiac chambers. Postmortem cardiac blood ethanol concentration as well as postmortem femoral venous blood ethanol concentration may usually reflect blood ethanol levels at the time of death when putrefaction is not evident. However, special attention is required for cases that received CPR after massive bleeding.

Adult↗
PubMed2005

Postmortem serum protein S100B levels with regard to the cause of death involving brain damage in medicolegal autopsy cases.

The protein S100 is an acidic calcium-binding protein, and the subunit S100B is the most abundantly found in the brain. The aim of the present study was a comprehensive analysis of serum S100B levels in medicolegal autopsy cases (within 48 h postmortem, n = 283) including victims with head and non-head injuries and also non-injury fatalities with regard to the cause of death involving brain damage. The serum level was usually higher in the subclavian vein than in the right heart and external iliac vein, and the lowest in the left heart blood, showing no significant postmortem influence. The serum level in the right heart and subclavian vein was markedly higher for acute deaths from head injury and asphyxiation due to neck compression (strangulation and hanging), and moderately and mildly elevated for other blunt and sharp instrument injury cases, respectively. For head injury, the serum levels were lower for subacute deaths than for acute deaths. These observations suggest that the elevation of serum S100B may mainly be caused by leakage following massive brain damage due to injury and cerebral hypoxia/ischemia, and in part by systemic hypoxic/traumatic tissue damage, depending on the survival time.

Adolescent↗
PubMed2005

Weight ratio of the lungs and pleural effusion to the spleen in the diagnosis of drowning.

An accurate and reliable method of diagnosing death by drowning is an important requirement in forensic autopsies. In this study, we compared the weight ratio of the lungs and pleural effusion to the spleen for 55 cases of drowning (37 males, 18 females), 36 cases of mechanical asphyxiation (16 males, 20 females), and 26 cases of acute cardiac death (19 males, 7 females). In the case of the males, there were significant differences in the weight of the spleen and the total weight of the lungs and pleural effusion between drowning and the other causes of death; however, there was no such significant difference in the females. We observed significant differences in the lungs and pleural effusion/spleen weight ratio between drowning and the other causes of death for both sexes. Therefore, these findings suggest that the ratio may be a useful index to accurately diagnose death by drowning, while ruling out mechanical asphyxiation and acute cardiac death in forensic autopsies.

Adolescent↗
PubMed2005

Postmortem stability of pituitary hormones in the human adenohypophysis.

The hypophysis is embedded in the fossa at the base of skull, having important functions in the hormonal system. The present study investigated its postmortem morphological changes and the stability of adenohypopyseal hormones. The pituitaries were collected at autopsy 6 h to 20 days postmortem and were studied by histology, immunohistochemistry and electron microscopy. To avoid the influence of prolonged brain hypoxia or swelling, subjects who survived not longer than 12 h were examined. Histological changes were seen in the nucleus 6 h after death, followed by cytoplasmic changes, and the cell shapes were hardly identifiable 7 days postmortem. Electron microscopy revealed evident ultra-structural changes 6 h postmortem, involving rough endoplasmic reticulum, Golgi complexes, mitochondria, nuclei and cell membranes. However, secretory granules remained well preserved 7 days postmortem. Immunostaining showed positivities for growth hormone, prolactin, adenocorticotropic hormone, luteinizing hormone and thyroid-stimulating hormone up to 15 days after death. These findings suggest the usefulness of immunohistochemical investigation of the adenohypophysis for estimating the time of death and endocrinologic evaluation in decomposed cadavers.

Adolescent↗
PubMed2005

Age-dependent increase of clusterin in the human pituitary gland.

Clusterin is a glycoprotein known to play various physiological roles including complement activity, amyloid binding activity in Alzheimer disease, as well as binding with heat shock proteins and abnormal prions. The present study immunohistochemically investigated the expression of clusterin in the human pituitary gland in subjects of 10-88 years of age (n=173). Causes of death were blunt injury (n=35), sharp injury (n=15), poisoning (n=11), drowning (n=14), fire fatalities (n=28), asphyxiation (n=15), hypothermia (n=7), hyperthermia (n=3), and natural diseases (n=45). Clusterin was detected in mixed cell follicles and the anterior lobar parenchymal cells. The area occupied by cells positive for clusterin were measured, and the ratio to the whole area of the anterior lobe (% clusterin-positive cell area) was estimated. There was a good correlation between the age of the subjects in years and the % clusterin-positive cell area in the anterior lobe of the pituitary gland (r=0.736, P<0.01). Relationships between % clusterin-positive cell and gender, cause of death, and survival time were insignificant. These findings indicate an age-dependent accumulation of clusterin in the pituitary gland, which may be related to the aging of endocrine systems.

Adolescent↗
PubMed2005

Gene expression of cytokines and growth factors in the lungs after paraquat administration in mice.

It is well known that the intake of paraquat (PQ), an herbicide, causes severe lung injury at chronic phases. We examined the intrapulmonary gene expression of cytokines and growth factors after PQ administration. To induce lung injury, C57BL/6 mice were intraperitoneally injected twice a week with 20 mg/kg of PQ. Histopathologically, at the early phase, lots of alveolar spaces contained edematous fluid. At 3 weeks after PQ challenge, a marked thickening of the alveolar walls with the accumulation of macrophages and T cells was found. Azan staining revealed the patchy distribution of collagen accumulation, indicating pulmonary fibrosis. Consistently, intrapulmonary hydroxyproline contents were significantly elevated, compared with the controls. Semi-quantitative RT-PCR analysis demonstrated that the gene expression of tumor necrosis factor-alpha and monocyte chemoattractant protein-1 were significantly increased at 3 weeks after PQ challenge compared with the controls. The mRNA expression of macrophage inflammatory protein (MIP)-1alpha and MIP-2 was significantly enhanced at 1 and 2 weeks after PQ treatment, respectively. Moreover, PQ-treated mice showed enhanced gene expression of fibrogenic growth factors such as transforming growth factor-beta, platelet-derived growth factor-A, acidic fibroblast growth factor, and hepatoctyte growth factor at 2 and/or 3 weeks after PQ challenge. The synergistic effects of these molecules are presumed to cause pulmonary fibrosis due to PQ challenge.

Animals↗
PubMed2005

Immunohistochemical distribution of S-100 protein in the cerebral cortex with regard to the cause of death in forensic autopsy.

S-100 protein (S100) is an acidic calcium-binding protein, which is abundantly found in the brain. The aim of the present study was to investigate the immunohistochemical distribution of S100 in the cerebral cortex in forensic autopsy cases with regard to the cause of death and relationship to its serum levels. The cases (n = 286, 3-48 h postmortem) included fatal head injuries (n = 89), acute death from other blunt injuries (n = 29), sharp instrument injuries (n = 20), asphyxiation (n = 29: strangulation/hanging, n = 22; aspiration, n = 7), drownings (n = 22), fire fatalities (n = 68), cerebrovascular diseases (n = 9) and acute myocardial infarction/ischemia (AMI, n = 20). S100-immunopositivity was mainly observed in the gliacytes, in part, in the neurons and myelins. For S100B-immunostaining, the gliacytes and myelins were positive, whereas the neurons were negative. Positivity in astrocytes was dependent on the cause of death, and was significantly lower in acute deaths due to strangulation/hanging and drowning, and mildly low in those due to injuries. Positivity in neurons and myelins was frequently observed in delayed head injury deaths and fire fatalities, showing an inverse relationship with the positivity in astrocytes in head injury cases. For cases of acute death, there was an inverse relationship between S100-positivity in the astrocytes and the serum S100B level. These observations suggest that astrocytes are more rapidly and severely injured than neurons during fatal brain damage, thereby causing an elevation in the serum S100B level.

Adolescent↗