Complementary roles for scavenger receptor A and CD36 of human monocyte-derived macrophages in adhesion to surfaces coated with oxidized low-density lipoproteins and in secretion of H2O2.

Oxidized low-density lipoprotein (oxLDL) is considered one of the principal effectors of atherogenesis. To explore mechanisms by which oxLDL affects human mononuclear phagocytes, we incubated these cells in medium containing oxLDL, acetylated LDL (acLDL), or native LDL, or on surfaces coated with these native and modified lipoproteins. The presence of soluble oxLDL, acLDL, or native LDL in the medium did not stimulate H2O2 secretion by macrophages. In contrast, macrophages adherent to surfaces coated with oxLDL secreted three- to fourfold more H2O2 than macrophages adherent to surfaces coated with acLDL or native LDL. Freshly isolated blood monocytes secreted little H2O2 regardless of the substrate on which they were plated. H2O2 secretion was maximal in cells maintained for 4-6 d in culture before plating on oxLDL-coated surfaces. Fucoidan, a known ligand of class A macrophage scavenger receptors (MSR-A), significantly reduced macrophage adhesion to surfaces coated with oxLDL or acLDL. Monoclonal antibody SMO, which blocks oxLDL binding to CD36, did not inhibit adhesion of macrophages to oxLDL-coated surfaces but markedly reduced H2O2 secretion by these cells. These studies show that MSR-A is primarily responsible for adhesion of macrophages to oxLDL-coated surfaces, that CD36 signals H2O2 secretion by macrophages adherent to these surfaces, and that substrate-bound, but not soluble, oxLDL stimulates H2O2 secretion by macrophages.

Mechanisms that regulate homing function of progenitor cells in myocardial infarction.

Cell based therapy has become a new and attractive option for the treatment of cardiac disease and heart failure. Although it has been demonstrated in vitro and in vivo that differentiation of non-differentiated cells (progenitor cells) into cardiomyocytes is able even in adult hearts the potential use of such transdifferentiation processes is limited by the small number of cells that home and engraft in the myocardium and complete the transdifferentiation process. Therefore, cell recruitment to the damaged heart is a major challenge to improve any cell based therapy. This process requires homing and engraftment of stem or progenitor cells. Major strategies to improve stem or progenitor cell homing are based on an improvement of stem or progenitor cell mobilization from the bone marrow. Strategies that have been shown to be successful are those that use granulocyte colony-stimulating factor (G-CSF). But although cell mobilization was indeed successful no major impact on hemodynamics was found. Alternatives are therefore needed and experimental studies use parathyroid hormone, statins, erythropoietin, and others in addition to or as an alternative to G-CSF. Although each of these procedures does have an impact on cell mobilization and homing none of these studies has provided a direct evidence that a major improvement on top of standard pharmacological therapy can be expected from such strategies. In conclusion, improvement of stem cell homing is a major challenge in the development of successful cell based therapies but not yet improved to a clinical relevant status. The underlying concepts of different strategies will be discussed here.

Factors and circumstances influencing the development of hemorrhages in livor mortis.

Petechial hemorrhages or ecchymoses in the skin of the face and/or in eyelids and/or conjunctivae are one important feature in postmortem diagnosis of lethal strangulation. On the other hand, petechial bleedings can occur in various causes of death, especially in cases of neck or thoracic compression, they can occur in acute cardiac failure, as a result of blood or skin diseases or as a postmortem phenomenon. The focus of this investigation (retrospective study of 279 corpses, found initially in a prone position or some other face down position) was to analyse the frequency of postmortem (hypostatic) hemorrhages and factors which may influence their development. Petechial hemorrhages in livor mortis in the skin of the trunk and extremities were found in 110 cases (39%). The frequency ranged from 41% in the side position and 44% in the kneeling position to 50% in the prone position. Increasing intensity of livor mortis resulted in an increasing frequency of hemorrhages, up to 59%. In cases with a body-mass-index (BMI) of more than 26 the frequency of hemorrhages increased up to 64%. In cases without livor mortis when the corpses were found as well as in cases with complete movement of livor mortis after turning the corpses, no hemorrhages were found. If hypostasis was partly or completely fixed, the rate of hemorrhages increased up to 50%, without additional increase in longer postmortem intervals. Obviously postmortem petechiae develop neither very soon nor days after death, but within a period of several hours after death.

Pretreatment with PKC activator protects cardiomyocytes against reoxygenation-induced hypercontracture independently of Ca2+ overload.

OBJECTIVE: Although several studies have shown that activation of protein kinase C (PKC) plays an important role in protection through ischemic preconditioning, little is known about the effects of direct PKC activation on the course of ischemia-reperfusion injury. The aim of this study was to analyse the effects of a pretreatment with the PKC activator 1,2-dioctanoyl-sn-glycerol (1,2DOG). METHODS: Isolated adult Wistar rat cardiomyocytes were exposed to 80 min of simulated ischemia (anoxia, pHo 6.4) and 20 min of reoxygenation (pHo 7.4). Cytosolic Ca2+ (fura-2), cytosolic pH (BCECF), Mg2+ (Mg-fura-2), lactate and cell length were measured and compared between control cells and cells treated with 20 mumol/l 1,2DOG before anoxia (10 min treatment and 10 min wash out). RESULTS: 1,2DOG-pretreatment delayed the time to extreme ATP depletion, but had no effect on lactate production and cytosolic pH. The accumulation of cytosolic Ca2+ was markedly accelerated in pretreated cells that developed rigor shortening, but reoxygenation-induced hypercontracture was significantly reduced. 1,2DOG, therefore, completely abolished Ca(2+)-dependence of hypercontracture. The effects of pretreatment were fully abolished with 1 mumol/l bisindolylmalcimide (PKC inhibitor). We conclude that PKC preactivation leads to (1) reduction of energy demand, (2) acceleration of Ca2+ overload during anoxia and (3) prevention of reoxygenation-induced hypercontracture independent of anoxic changes in cytosolic Ca2+ and pH.

Drug-antibody conjugates with anti-HIV activity.

Human immunodeficiency virus (HIV)-specific peptide antibody-brefeldin A conjugates and antibody-glaucarubolone conjugates directed to cell surface viral glycoprotein epitopes were prepared and tested for antiviral activity. A selective response was observed both on survival of cell lines permanently infected with lentiviruses and on HIV infectivity. With human peripheral blood mononuclear cells (PBMCs), the conjugate also was effective in reducing virus titers. The effectiveness of an HIV-specific peptide antibody-brefeldin A conjugate was enhanced by combination with 3'-azido-3'-deoxythymidine (AZT) and was effective against AZT-resistant isolates in combination with AZT. The conjugates reduced virus production in MOLT-4 cells and in HIV-1-infected PBMCs without affecting the viability of uninfected cells.

Lethal hemoptysis caused by biopsy injury of an abnormal bronchial artery.

A 62-year-old man with a long history of lung disease developed atelectasis of the right middle lung lobe, caused by a protrusion in the wall of the middle lobe bronchus. A biopsy was performed in the suspicious region. This was immediately followed by massive arterial bleeding into the airways and complicated by cardiac arrest soon after. The bleeding could not be controlled by nonsurgical treatment; the patient died 24 h after the complication because of pulmonary insufficiency. Autopsy revealed the bleeding to have been caused by a biopsy injury of a bronchial artery that had run superficially in the bronchial mucosa and had produced the intrabronchial protrusion. Several other abnormal intrabronchial arteries were found peripherally in this lung.

Effect of the quassinoids glaucarubolone and simalikalactone D on growth of cells permanently infected with feline and human immunodeficiency viruses and on viral infections.

Growth of Crandall feline kidney cells permanently infected with feline immunodeficiency virus was inhibited by the anti-cancer quassinoid glaucarubolone whereas growth of uninfected cells was not inhibited. Similar results were obtained for human MOLT-4 cells infected with HIV-1. The results suggest that cell lines permanently infected with either the feline or the human lentivirus exhibit growth response characteristics to the quassinoids in common with other cell lines malignantly transformed. In addition the quassinoids may delay viral infection suggesting some commonality between the mechanism responsible for inhibition of the growth of the transformed phenotype and viral infection.

Congestion bleedings of the face and cardiopulmonary resuscitation--an attempt to evaluate their relationship.

OBJECTIVES: Are any other factors besides the factor "cause of death" involved in the development of petechial hemorrhages (PET) of the head? The significance of the cause of death is well known, other factors have been rarely investigated in medical literature. Do they include cardiopulmonary resuscitation (CPR), as has been claimed in several forensic publications? MATERIALS AND METHODS: (a) 473 consecutive autopsy cases (without strangulation) evaluated by one examiner, which were appropriate for this investigation; (b) analysis of 181 cardiac deaths (investigated by all physicians of our institute). RESULTS: Petechiae were found in 13.3% of all cases and were clearly dependent on the cause of death, up to 20% were found in burn victims, intensive-care patients and cardiac fatalities. Petechiae were more frequently observed in the middle age groups (>20%) than in old persons (<10%). The number of PET cases increased with body mass but was lower in extremely obese persons, a greater number of cases with PET was also observed with increasing heart weight. PET were observed in 11% of the deaths without CPR compared to 19% with CPR. This difference was predominantly caused by the subgroup "acute coronary death", especially if victims younger than 60 years were considered, whereas in many other causes of death no difference in the prevalence of PET with or without CPR could be observed. CONCLUSION: Besides the cause of death, other factors (age, body mass and possibly even heart weight) influence the development of petechiae. The hypothesis that CPR alone produces PET is not confirmed by our experience.

Detection of ruptured cerebral bridging veins at autopsy.

Subdural hematomas (SDH) are a frequent autopsy finding in victims of closed head injuries. About 2/3 of the cases in our own massive SDH series had brain contusions as a cause of bleeding. In about 1/3 of the cases, SDH resulted from ruptured vessels of the brain surface in the absence of contusions. Our experience has shown that such an "isolated" SDH may be expected in about 5% of fatal blunt head injuries. A technique is presented to detect bridging vein leakage before there is any risk of artificial damage. After sawing through the skull in the fronto-occipital plane and cutting through the upper half of the brain together with the calvarium, approximately 5-10 ml of barium sulfate are instilled into the superior sagittal sinus by a balloon catheter at low pressure, and X-rays are taken. If there is extravasation, the rupture is visible before further preparation.

Homicidal and suicidal ligature strangulation--a comparison of the post-mortem findings.

Suicides by ligature strangulation are rare events. In Berlin (3.5 million inhabitants; ca. 500 suicides per year) approximately one case per year occurs. Here, we present the main findings of 19 cases investigated between 1978 and 1998, compared to 47 cases of homicidal ligature strangulation. Two of the 19 suicidal victims had single fractures of the upper thyroid horns and one victim a fracture of a lower thyroid horn; other types of laryngohyoid injuries were not observed. In the homicidal series, the laryngohyoid structures were unaffected in 26 cases (12 of these victims were children or adolescents), single thyroid horn fractures were present in three cases and more significant injuries in 18 cases. Macroscopic bleedings of the laryngeal muscles were found in 12 victims of the homicidal group and in none of the suicidal. Bleedings in the neck muscles seldom occurred in suicides. According to these findings, the laryngohyoid injuries can be helpful in the differentiation of suicide from homicide, if more than a single thyroid horn fracture or a laryngeal soft tissue trauma is present.

Hemorrhages of the tongue in the postmortem diagnostics of strangulation.

Hemorrhages of the tongue can be useful for the diagnosis in cases of lethal neck compression. The reported frequencies of tongue bleedings in the literature in cases of suicidal hanging range from 0 to 14% and in homicidal strangulation (str.) from 5 to 37%. This study gives a summary of the incidence and intensity of tongue bleedings in cases of homicidal str. by ligature, manual str., suicidal str. by ligature and suicidal hanging. In 25% of all homicides significant or massive hemorrhages of the tongue were detected. In contrast to this, in suicidal hanging the tongue was unaffected in 95%. The causes of massive hemorrhages here (2%) could be explained by an "abnormal" position of the loop.

Lethal suicidal intoxication with propafenone, after a history of self-inflicted injuries.

Report of a suicidal mono-intoxication with the class IC antiarrythmic drug propafenone. A 20-year-old female physician's assistant secretly ingested the substance (presumably 20 tablets per 300 mg) about 4-6 h before her death, and in the interim remained under the supervision of her physician. An ECG taken about 1/2-2 h after ingestion showed widening of the QRS complex and signs of an acute load of the right ventricle; the clinical symptoms were nausea, vomiting and hypotonia. After about 4 h without serious symptoms acute loss of consciousness and cardiac failure occurred, resuscitation efforts remained unsuccessful. At autopsy propafenone was found in blood (12 micrograms ml-1), liver (60 micrograms g-1) and cardiac muscle (11 micrograms g-1).

Medical studies and training duration for forensic pathologists in Germany.

This article gives a short overview of medical education in Germany. The legal basis and organization of the medical studies program as well as the course in forensic medicine and the training duration for forensic pathologists are described.

Demonstration and interpretation of bridging vein ruptures in cases of infantile subdural bleedings.

Report of two cases of lethal infantile subdural bleedings (SDB). Bridging vein (BV) ruptures were directly proven as the source of the (minimal) SDB by a postmortem X-ray. In the controversial discussion concerning the causes of infantile SDB, proof of the occurrence of several BV ruptures is seen as an important sign of a trauma of significant degree. Although infantile SDB undoubtedly can result from accidental as well as intentional injuries, and therefore, the SDB itself does not allow far-reaching conclusions as to the cause of injury, the presence of several BV ruptures combined with an SDB of insignificant volume, in an infant dead or in a deep coma on clinical presentation, is not compatible with the supposition of a minor fall as the cause. We have not observed such findings as the result of a minor accidental event for more than 15 years.

"Hidden" laryngeal injuries in homicidal strangulation: how to detect and interpret these findings.

These studies were designed to demonstrate that the usual method of laryngeal dissection carries a significant risk of overlooking important laryngeal injuries. Formalin-fixed hyoid-larynx complexes were prepared in a prospective forensic study involving 191 cases of homicidal strangulation, 1984 to 1997. The basic steps of the applied method included: complete resection of the thyroid cartilage, a horizontal incision through the cricoid cartilage before opening the larynx dorsally, inspection of the laryngeal joints, and incisions of the laryngeal muscles. Using this procedure allowed us to detect the following injuries, which otherwise would have been destroyed or overlooked: (a) 17 incomplete fractures restricted to the dorsal surfaces of the thyroid laminae and 10 incomplete or non-dislocated fractures of the cricoid cartilage. In 7 cases, such a "hidden" fracture was the only laryngeal injury resulting from neck compression. (b) Extensive laryngeal muscle hemorrhages, especially of the vocal folds, were found in almost half of all cases, more rarely in strangulation by ligature and more frequently in manual strangulation. Gross hemorrhages were the decisive local laryngeal finding in 19 cases. (c) Laryngeal joint injuries (bleedings) were found in 18% to 52% of the different strangulation types. (d) Hemorrhages of the laryngeal mucosa were common findings that occur in about 60% of all cases; only in rare cases do such bleedings have a special diagnostic value. The quantity and significance of findings obtainable from complete preparation clearly document Camps's demand made in 1976 to dispense with the usual laryngeal dissection technique (dorsal scissor incision through the cricoid cartilage), at least in (questionable) strangulation cases.

Neck injuries caused by being run over by a motor vehicle.

We report a homicide involving the use of a motor vehicle and simulating a traffic accident. This observation was the reason for a retrospective analysis of neck injuries in victims of traffic accidents, in which a person has been run over (RO) by a motor vehicle. The autopsy material of two institutes from 1990-1996 was used. The following findings were obtained in 63 victims: laryngohyoid fractures (LH-fx): 10 cases (16%) with a clear difference between the institutes (22% versus 7%). This resulted from examination with special regard to such injuries in many cases at one of the institutes, whereas only autopsy reports were taken retrospectively from the other institute. Five of these cases had suffered only minor LH-fx (as seen frequently in strangulation), although extensive run over (RO) injuries of the other cervical tissues were present. All LH-fx were caused by direct compression of the neck; in eight of the cases they were combined with mandibular fractures. Petechial hemorrhages (petH) at the eye lids/conjunctivae were seen in 19 cases (30%); 16 of these were related to thorax RO injuries, three to abdominal RO only. Four cases involved LH-fx, petH as well as cervical skin lesions and additional cervical soft tissue hemorrhages. Interpretation can be extremely difficult with this combination of findings if the character of the event cannot be established as accidental beyond doubt on the basis of the circumstances.

[Healed fractures of the larynx and lingual bone in forensic autopsy]. / Abgeheilte Frakturen von Kehlkopf und Zungenbein im forensischen Obduktionsgut.

Laryngohyoid fractures are a frequently investigated matter, especially in the forensic literature. On the other hand, there are only very few (old) forensic reports of such fractures in survived cases. However, healed fractures are not seldom found in forensic autopsies: In a personal series of 1160 forensic autopsies (adult persons) a careful dissection of the laryngohyoid complex was done by 1 investigator. Only a macroscopic examination of the cartilages was carried out; radiographs and histological slices were not regularly made. Therefore only a part of existing old fractures is detectable; for example, healed fissures are not visible with this simple method. Furthermore, asymmetries of the thyroid laminae cannot be declared as posttraumatic without additional examination, because this condition is described as possible anatomic anomaly. From that, the injury frequencies presented here only mark the lower threshold of the existence of such findings. Healed fractures of the upper thyroid horns and the major hyoid horns can be easily detected even in a routine examination, if healing resulted in a fixed dislocation or apposition of bone surrounding the former fracture site. In the present series, this was the predominant localization of old fractures: the upper thyroid horns (43 cases), followed by the hyoid cornua (12 cases), a combination of both sites (5) and cricoid fractures (5). Of the total 65 healed fractures (5.6%), 35 were found in the group of 290 chronic alcoholics (12.1%) and only 3.4% in non-alcoholics. In the subgroup of middle-aged alcoholics, the fracture rate increased up to 19%. However, this group did not present a higher rate of fresh laryngohyoid injuries (not related to strangulation) than the other cases. The old fractures probably resulted from minor "daily" injuries (like falls), which are common, especially in chronic alcoholics. The frequency of such findings should be in mind if an apparently fresh fracture, found in an actual autopsy, should be related to the cause of death: there is a real chance, that this fracture occurred prior, and without causal connection to the factors resulting in death. Therefore a histological examination of the age of this finding is necessary. Cricoid fractures are quite uncommon, except in serious external neck trauma. In 1 of our 5 cases, this fracture was caused by repeated cruelty, finally resulting in death.

[Value of laryngeal injuries as evidence]. / Zum Beweiswert von Kehlkopfverletzungen.

Report of two cases: Violations at soft parts of the neck and at larynx pointed to an attack against the throat by strange hand, but local vital reactions showed that the attack had been survived. A 54-year old woman had little excoriations in the skin of the neck, the right great horn of hyoid bone and the left upper process of thyroid cartilage were broken, the fractures were bloodshot. No petechiae in skin and conjunctivitis. Death occurred from a suicidal poisoning of soporifics. The attack against the throat (throttling by hand, conceded by the culprit) had been survived. That was recognizable on a clear cell-reaction in the bleedings within the muscles of the neck. A 87-year old man was found in his rummaged home. Violations of the skin in the face pointed to attacks against mouth and throat; additional maltreatment by gagging and fettering (in the meantime untied) could be supposed. -- Histological examination showed advanced cell-reaction in the wounds, partially even intracellular iron pigment; there was a superposition of old and recent injuries, but even the recent had been survived many hours. The situation of finding the corpse, bleedings inside the iliac muscles, little hemorrhages in the mucosa of the stomach and acetone in blood and urine pointed to a death lately from cold. But complete clearing-up of the case was not yet possible.

[Injuries to laryngeal articulations in strangulation and throttling]. / Verletzungen der Kehlkopfgelenke beim Würgen und Drosseln.

During the consecutive examination of the larynges of (81) deceased persons (strangulation by hand and/or ligature, blunt force against the neck, from other persons), the cricothyroid and cricoarytenoid-articulations were prepared and opened, and capsular or articular cavity hemorrhaging was found in 30% of all cases; a clear distinction being found between effect of manual strangulation (43%) or blunt trauma (44%) on the one hand and strangulation by ligature alone (9%) on the other. The greater the extent of injury to the cartilages the more articular lesions are found as an expression of the mechanical load on the entire laryngohyoid structure.

[Delayed death following strangulation (hanging)]. / Spättod nach Strangulation (Erhängen).

6 cases of suicide by hanging with delayed death are presented in tables laying out the circumstances and the findings. All the persons involved remained continuously unconscious after initially successful reanimation; two were at this time already dead. The survival times lay between 18 hours and 4 days. In two cases intravital vertebral death occurred. In the other cases death was due indirectly to hypoxial brain-damage which had led to a greater or lesser extent to elective parenchymal necrosis. In 3 of 4 histologically investigated cases there was evidence of extensive cell destruction in the PCA muscle which was, however, not specifically related to the process of hanging since lesions of this nature also occur with other cases of delayed death after reanimation. In 7th case the problems encountered in compiling an expert opinion are discussed in cases where there are no corresponding procedural details. A 44 year old man died 3 weeks after having been submitted to bondage in a brothel at his own request (including binding of the neck). Compression of the throat with emphasis on one side was evident resulting in extensive ganglial cell damage in one half of the cerebral cortex.