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PubMed2003

Post-mortem changes in calmodulin binding proteins in muscle and lung.

Estimation of post-mortem interval (PMI) remains an elusive issue in forensic investigations. In this study, we examined the possible use of calmodulin (CaM) binding proteins (CaMBPs) as indicators of PMI. Whole CaMBP populations from homogenized rat lung and rat skeletal muscle removed at 0, 24, 48 and 96 h post-mortem at 21 degrees C were detected by the calmodulin binding overlay technique (CaMBOT) using 35S-VU1-CaM and visualized by autoradiography. CaMBOT showed that, in both tissues, the CaMBP population remained relatively stable for up to 96 h post-mortem with the exception of a single approximately 200 kDa CaMBP that increased in 24 h post-mortem samples then showed decreasing amounts at subsequent times. Immunoblot analysis of the specific CaMBPs, Ca(2+)/CaM-dependent kinase II (CaMKII), calcineurin A (CNA), myristoylated alanine-rich C-kinase substrate (MARCKS) and inducible nitric oxide synthase (iNOS) were done on lung tissue samples. CaMKII levels did not change appreciably over the 96 h PMI examined. In contrast to iNOS levels, which varied from sample to sample, CNA and MARCKS showed predictable patterns of change: the level of MARCKS decreased steadily in the 0-96 h post-mortem lung samples while CNA underwent a shift in mobility on SDS-PAGE by 24 h post-mortem before slowly decreasing in amount. The stability of CaMKII levels over 96 h was also seen in skeletal muscle tissue while CNA showed variable levels at 0, 48 and 96 h with the presence of the rapidly migrating band at 24 h. These patterns of change in CaMBPs provide some insight into the post-mortem changes in calmodulin-mediated signaling components in lung and skeletal muscle and support the further study of CNA and CaMKII as potential markers for estimating short- and long-term PMIs.

Analysis of Variance↗

PubMed2006

Could NF-kappaB and caspase-3 be markers for estimation of post-interval of human traumatic brain injury?

BACKGROUND: Previous studies in a rat model showed expressions of caspase-3 (p20) and NF-kappaB (p65) in different time after TBI. We test the activation changes of caspase-3 and NF-kappaB of human specimen after traumatic brain injury (TBI) in this study to see whether they could be markers to estimate post-intervals of TBI for forensic practice. MATERIAL AND METHODS: Caspase-3 (p20) and NF-kappaB (p65) immunohistochemical staining were performed on 39 TBI specimens (dead from TBI) and eight controls (dead from none TBI reasons) grouped 0, 12, 24, 48, 72, 168, 264 and 480 h. Positive cells were counted and image analysis techniques were used to determine the morphological changes in each group. RESULTS: Immunohistochemical analysis showed a marked induction of caspase-3 (p20) in every injured group compared with normal controls; each positive group has it's morphological character with variant positive cell count. NF-kappaB (p65) did not showed markedly change compared with normal controls until 168 h after TBI and almost all nerve cells were stained by NF-kappaB (p65) in 264 and 480 h groups. CONCLUSION: NF-kappaB (p65) could contribute to estimate delayed death cases after TBI for forensic applications. Caspase-3 (p20) estimating post-TBI intervals are dubious but could suggest TBI exists.

Adult↗

PubMed1975

[Quantitative investigation of the amino acid levels in putrefying liver (author's transl)].

To elucidate the post-mortem proteolysis we investigated the behaviour of the free amino acids in rotting liver homogenates by means of two-dimensional thin-layer chromatography and reflection densitometry. The post-mortem alterations of the amino acid levels are characterized mostly by a two-phase increase with maxima at the end of the first and after the third week. A few amino acids showed only little changes hardly above the physiological intra-cellular levels. Anaerobic rat experiments and experiences with antibiotic treatment give rise to the supposition that increasing amino acid concentrations indicate a predominance of proteolytic activities over aminoacid catabolism. In the beginning autoenzymes are prevalent, while afterwards bacterial proteases probably predominate. The transient regression about the second and thrid week is interpreted as the effect of a temporarily intensified amino acid degradation by foreign enzymes (maximal bacterial growth).

Amino Acids↗

PubMed1992

Ultrastructural observations on autolytic changes of atrial specific granules in pig heart.

The progressive autolytic changes of atrial specific granules (ASGS) up to 24 hours after death were studied electron microscopically. Tissue samples taken from the left auricular myocardium of four pig hearts at 0, 0.5, 1.5, 3.5, 6, 12 and 24 hours post mortem were fixed for 24 hours by immersion in 2.5 per cent cacodylate buffered glutaraldehyde (pH 7.4). Two pig hearts were kept at 20 degrees C and two at 4 degrees C. Although typical autolytic changes were seen in many organelles, no remarkable modifications in aspect, distribution, content and diameter of ASGS were observed in all samples. The results of this study indicate that ASGS are not morphologically affected by the autolytic processes, at least during the first 24 hours. Hence, the electron microscopic evaluation of ASGS does not need a particular fixation method such as perfusion. This may eliminate technical problems when slaughterhouse material is used.

Animals↗

PubMed1994

Renal osteodystrophy and dialysis artifacts as indicators of identification.

Evidence of rare disease may be useful in the identification of unknown remains. Osteodystrophy is a condition commonly associated with chronic renal failure. The presence of renal osteodystrophy and medical artifacts associated with the treatment of kidney failure in human remains may provide information about the individual's medical history. Currently, there are more than 100,000 patients with end stage renal disease in the United States receiving dialysis treatments to replace kidney function, and hundreds of thousands more who have significant chronic kidney failure and are not yet treated with dialysis. Chronic renal failure frequently leads to disorders in the metabolism of vitamin D, calcium, and parathyroid hormone which are extremely difficult to correct. At least three patterns of skeletal change may result: osteitis fibrosa, characterized by increased bone remodelling, increased osteoclastic activity, peritrabecular fibrosis, and a normal mineralization pattern; osteomalacia, with a markedly increased osteoid surface and volume, and impaired mineralization; and mixed osteodystrophy, with increased bone remodelling and moderately impaired mineralization. While these bone changes may not have any clinical manifestations in most patients, they can been seen radiographically and histologically. This evidence may be useful in identifying remains which have been skeletonized, burned, decomposed, or dismembered. Medical artifacts associated with dialysis treatment, such as catheters, polytetrafluoroethylene grafts, and evidence of surgical procedures, are also useful for identification.

Adult↗

PubMed1990

Spontaneous DNA breaks in the rat brain during development and aging.

The level of spontaneous DNA breaks in nuclei isolated from the cerebral cortex of rat embryos at 12, 15 and 19 days of gestation, and from cerebral cortex and cerebellum of 24-day-, 6-month- and 36-month-old rats was measured by alkaline elution. A constant low level of DNA breaks was found in brain DNA during development from an embryo at day 12 of gestation to a 24-day-old rat. During aging the level of DNA breaks remained at the same low level, as shown by comparing nuclei from the cerebral cortex and cerebellum of 6- and 36-month-old animals. By contrast, an almost 2-fold increase in the level of DNA breaks was observed in rat liver nuclei between 6 and 36 months of age, confirming our earlier findings on isolated liver cells. Although there were no changes in the level of DNA breaks in rat brain during development or during aging, breaks accumulated rapidly post mortem. The rate of this process was not age-dependent. Our data suggest that the level of spontaneous DNA breaks in the brain is not likely to be of fundamental importance in the complex cellular alterations associated with brain development and aging.

Aging↗

PubMed1988

Reductions in [3H]nicotinic acetylcholine binding in Alzheimer's disease and Parkinson's disease: an autoradiographic study.

In Alzheimer's disease (AD) and Parkinson's disease (PD), dysfunction in the basal forebrain cholinergic system is accompanied by a consistent loss of presynaptic cholinergic markers in cortex, but changes in cholinergic receptor binding sites are poorly understood. In the present study, we used receptor autoradiography to map the distribution of nicotinic [3H]acetylcholine binding sites in cortices of individuals with AD and PD and matched control subjects. In both diseases, a profound loss of nicotinic receptors occurs in all cortical layers, particularly the deepest layers.

Aged↗

PubMed1994

Post-mortem limitations of body composition analysis by computed tomography.

Few of the indirect methods for measuring body composition have every been validated against direct human cadaver evidence. Computed tomography (CT), like NMR, has proved to be an important diagnostic tool and they appear to be the techniques of the future for body composition studies. The purpose of the present study (Cadaver Analysis Study III), undertaken at the Vrije Universiteit Brussel in a joint venture with the University of Göteborg, Sweden, was to validate tomographic measurements of volumes and areas from different tissues using data from CT-scanning of unembalmed deep-frozen cadavers and data collected by dissection of the same cadavers. Six Belgian adults were extensively measured and dissected. The body was divided into several slices for comparison of the CT image with photography of the same slice and comparison of tissue-volumes per segment for the whole body. Due to post-mortem changes and the frozen state of the cadavers, the CT measurements were greatly affected by artefacts disturbing adipose tissue (AT) and muscle area determinations. Only the bone area measurements were similar between the two techniques. However, when the volumes (per segment) of the same tissues were considered, no apparent difference was found between CT and dissection data for the muscle volume.

Aged↗

PubMed1979

Studies on the post-mortem fragmentation of myofibrils.

1. There was a close relationship between the fragmentation of myofibrils and the tension developed during post-mortem contraction of muscle. The extent of fragmentation was at its maximum when the sarcomeres attained a length of 2.0 to 2.2 micron. 2. The rate of fragmentation of myofibrils depended upon the calcium ion concentration within a range of 10(-5) to 2 x 10(-2) M, with a minimum at pH 6.5. The fragmentation of myofibrils free from muscle fibers was not affected by 10 mM iodoacetate, an irreversible inhibitor of calcium-activated factor (CAF). 3. Incubation of myofibrils with 10 mM CaCl2 caused the release of about 12% of the total myofibrillar proteins after homogenization. The protein solution contained little alpha-actinin, and considerable amounts of 54,000- and 76,000-dalton components which seem to originate from the Z-line. SDS-polyacrylamide gels of troponin prepared from the incubated myofibrils did not change with time of incubation. These findings are in contrast with the proteolytic degradation of Z-lines by CAF treatment, in which alpha-actinin and 87,000 dalton component are released. 4. These data directly demonstrate that the in vitro fragmentation of post-mortem muscle (i.e. duirng its conversion into myofibrils upon mechanical homogenization) is different from that induced by CAF. The possible role of calcium ions during in vitro fragmentation of myofibrils is discussed.

Animals↗

PubMed1989

Analysis of post mortem aqueous humour chemistry in the horse, with particular reference to urea nitrogen and creatinine.

The concentrations of several post mortem aqueous humour chemical constituents were compared with ante mortem serum chemical values in the horse. Urea nitrogen and creatinine values in post mortem aqueous humour were good predictors of ante mortem serum values. Aqueous humour urea nitrogen increased only slightly and creatinine did not change significantly for up to 24 h after death. Formulae were derived for calculating estimated ante mortem serum urea nitrogen and creatinine from aqueous humour values obtained after death. These results from normal horses identify analytes that are accurate predictors of ante mortem serum values. Determination of post mortem aqueous humour urea nitrogen of creatinine may assist in interpretation of the functional significance of equivocal histological lesions in the kidney.

Analysis of Variance↗

PubMed1994

[The ABO group system in tissues in advanced post-mortem changes].

Post-mortem changes and in particular putrefaction makes examination of the group properties of the ABO system in tissues difficult and even more so the interpretation of results. Hitherto traditionally used serological methods, i.e. the absorption elution test and the saturation method, do not always provide sufficient data for a final evaluation. The quoted examples from practice are in favour of a combination of the traditional serological approach (absorption elution test) and the immunohistological technique (indirect immunoperoxidase reaction).

ABO Blood-Group System↗

PubMed2005

Cortical glutamatergic markers in schizophrenia.

Post-mortem studies have yet to produce consistent findings on cortical glutamatergic markers in schizophrenia; therefore, it is not possible to fully understand the role of abnormal glutamatergic function in the pathology of the disorder. To better understand the changes in cortical glutamatergic markers in schizophrenia, we measured the binding of radioligands to the ionotropic glutamate receptors (N-methyl D-aspartate, [3H]CGP39653, [3H]MK-801), amino-3-hydroxy-5-methyl-4-isoxazole ([3H]AMPA), kainate ([3H]kainate), and the high-affinity glutamate uptake site ([3H]aspartate) using in situ radioligand binding with autoradiography and levels of mRNA for kainate receptors using in situ hybridization in the dorsolateral prefrontal cortex from 20 subjects with schizophrenia and 20 controls matched for age and sex. Levels of [3H]kainate binding were significantly decreased in cortical laminae I-II (p = 0.01), III-IV (p < 0.05), and V-VI (p < 0.01) from subjects with schizophrenia. By contrast, levels of [3H]MK-801, [3H]AMPA, [3H]aspartate, or [3H]CGP39653 binding did not differ between the diagnostic cohorts. Levels of mRNA for the GluR5 subunit were decreased overall (p < 0.05), with no changes in levels of mRNA for GluR6, GluR7, KA1, or KA2 in tissue from subjects with schizophrenia. These data indicate that the decreased number of kainate receptors in the dorsolateral prefrontal cortex in schizophrenia may result, in part, from reduced expression of the GluR5 receptor subunits.

2-Amino-5-phosphonovalerate↗

PubMedDate not supplied

[The tissue survival problem in using current methods of morphological study].

Time course of ultrastructural changes and RNA production in human brain and myocardial cells during the period between clinical and biological death is studied. Changes in the cytoplasm of brain neurons develop 4 hours after death, and 8 h post mortem the destructive changes in the cytoplasm and nuclei progress. During the period of 4-13 h post mortem the endotheliocyte nuclei are capable of producing RNA. Active production of RNA is on-going in the myocyte nuclei 4 h after death; the synthetic activity decreases by 8 h and by 13 h the synthetic activity in autopsy material is over.

Adolescent↗

PubMed2001

Percutaneous retrieval of the Tulip vena cava filter: feasibility, short- and long-term changes--an experimental study in dogs.

PURPOSE: To evaluate experimentally the retrievability of the Tulip inferior vena cava (IVC) filter in an in vivo study. Changes which accompany venous healing after filter retrieval were investigated. METHODS: In 12 dogs, 23 filters were inserted percutaneously into the lumbar and intrahepatic segments of the IVC. Two weeks (n = 21 filters) or 3 weeks (n = 2 filters) after insertion, filter retrieval was attempted through an 11 Fr coaxial retrieval sheath system placed via the jugular vein. Follow-up studies before and after filter retrieval included cavography, computed tomography and intravascular ultrasound of the IVC. Seven dogs were killed immediately after filter retrieval to confirm short-term changes of the IVC, and 5 dogs were killed 6 months after filter retraction to evaluate long-term changes of the IVC related to filter retrieval. Post-mortem examinations and histologic specimens of the IVC were obtained to evaluate caval wall abnormalities secondary to filter removal. RESULTS: All but one filter were successfully retrieved 2 weeks post-implantation. However, 3 weeks after insertion, filter retrieval was impossible. There were no complications caused by filter extraction. Follow-up studies after filter retrieval revealed no significant changes in the integrity, morphology or composition of the IVC and pericaval tissue. Histologic examination 6 months after filter retrieval revealed only flimsy fibrotic intimal plaques at the sites of former hook insertion. CONCLUSION: The Tulip filter allows percutaneous insertion and retrieval up to 14 days after insertion, suggesting that it may be useful for either permanent or temporary prophylaxis against pulmonary embolism.

Animals↗

PubMed1994

Synaptophysin gene expression in human brain: a quantitative in situ hybridization and immunocytochemical study.

Synaptophysin is a presynaptic vesicle protein. Its quantitative detection has become established as a molecular marker of synaptic density. We have studied synaptophysin messenger RNA in the neocortex, hippocampus and cerebellum using in situ hybridization histochemistry to see if the encoding transcript can be detected in post mortem human brain and to investigate factors which might influence its abundance. Synaptophysin was also measured immunocytochemically in the hippocampus. The level of synaptophysin messenger RNA expression was uniform in all neocortical areas examined. Strong correlations were found for the amount of synaptophysin messenger RNA between individual regions and between homologous areas in the two hemispheres. Synaptophysin messenger RNA declined with increasing age and prolonged post mortem interval. Synaptophysin immunoreactivity also reduced with age, as well as with duration of formalin fixation but not post mortem interval. Synaptophysin immunoreactivity correlated with the abundance of the messenger RNA in neurons within, and projecting to, each hippocampal subfield. Significantly greater synaptophysin immunoreactivity was seen in the left than the right CA4 and CA1 regions. These data show that quantitative detection of synaptophysin messenger RNA as well as synaptophysin itself can reliably be carried out in post mortem human brain sections. They are in keeping with other findings that synaptic density is relatively uniform through the neocortex and decreases with age. They also suggest a possible asymmetry of hippocampal synaptophysin expression. The level of synaptophysin messenger RNA paralleled that of synaptophysin immunoreactivity, indicating that changes in gene expression contribute to variations in the latter observed in aging and other situations. Detection of synaptophysin messenger RNA broadens the range of methods by which synaptic protein gene products can be studied and used as markers of synaptic density and synaptic involvement during physiological and pathological processes in human brain.

Adult↗

PubMed2002

The effect of maceration on the dental arches and the transverse cranial dimensions: a study on the pig.

The dimensional change of the dental arches and the transverse cranial dimensions were studied in the pig to gain information on cranial post mortem changes and thus improve the possibilities of comparison between modern and skeletal samples. Dental arch dimensions were registered in 17 pigs within 30 minutes after they had been killed. The following day, the skulls were registered on lateral and axial radiographs. The animals were prepared, and storage and preparation included freezing and treatment in hot water. The water temperature did not exceed 65 degrees C. After this process, the skulls were again registered on lateral and axial radiographs. The skulls were then allowed to dry for 2 weeks and the direct measurements were repeated. The dimensions showed shrinkage of between 0 and 3.3 per cent. The mandible showed a greater change transversally in the posterior region than the cranium, which may have been due to its shape. The more deviant values for dimensional change were probably due to technical errors and the shrinkage may be expected to vary from 0.3 to 1.7 per cent, with greater values occurring in the posterior transverse parts of the mandible. A differential shrinkage in the maxilla could not be excluded and the values varied between 0.3 and 1.9 per cent. The results indicate that the crania in skeletal samples can be expected to be 0.3-1.7 per cent smaller than in vivo.

Animals↗

PubMed1991

[Pattern of experimentally-induced subcutaneous hematoma].

Healthy persons were each injected with 1 ml autoblood into the subcutis of the inner upper arm. Following this, different objects were applied to each haematoma area and kept pressed on the area for one hour by means of a sphygmomanometer cuff (30 mm Hg), to establish whether this would lead to secondary shape or pattern changes of the haematoma. When the haematomata were compressed by shaped objects, there developed patterned blanched areas or intensity differences, respectively, becoming more marked in the course of several days in the living experimental persons. In corpses greater quantities of blood were injected (10 ml) and higher contact pressures applied (up to 180 mm Hg). In some cases a markedly delimited imprint reflecting the configuration of the object became discernible already one hour post injectionem.

Adult↗

PubMed1990

[Electrical impedance and time of death in the animal model].

To study passive electric parameters of the rat skeletal muscles in relation to the time of death, investigations were carried out using the impulse distortion method of Pliquett. This measuring method enables the evaluation of the electric rectangular impulse distorted by the biological tissues, which contains any essential information on the passive electric properties of the object to be measured. The passive electric parameters change in the studied time interval up to 79 h post mortem in a characteristic fashion. The linear correlation of the evaluated impulse parameters at the time of death is closest (r = 0.75) between 0.5-8.5 h post mortem, the scatter of single values in late postmortal periods is in part considerable. A further development of the measuring technique and standardization of the measuring method let the passive electric parameters appear to be appropriate magnitudes for estimating the time of death.

Animals↗