Multiple hemangiomas and hemangiomatosis--risk factors and outcome over an eight year period.

INTRODUCTION: Hemangiomas are the most common tumors of infancy. Multiple cutaneous hemangiomas may be associated with the presence of hemangiomas in inner organs. However, there is little data on the risk factors for organ involvement and the outcome of a large sample of patients. PATIENTS AND METHODS: patients with 3 or more cutaneous hemangiomas were evaluated with regard to patient characteristics, distribution of hemangiomas, results of radiological abdominal/cerebral imaging, clinical course, and therapeutic approach. We analyzed the risk factors for organ involvement and complications/outcome. RESULTS: The average gestational week at birth was 32.8; radiological imaging showed liver hemangiomas in 13.5% and mesenteric lesions in 1 (1.9%) but no cerebral lesions. Preterm infants (p=0.02) and patients with high numbers of cutaneous hemangiomas (p=0.02) were at higher risk of organ involvement. A life-threatening event occurred in 1 patient (1.9%). None of the patients died. CONCLUSIONS: Organ manifestation is relatively common in patients with multiple hemangiomas, complications are rare, but potentially life-threatening. We recommend abdominal imaging for patients with 3 or more hemangiomas, especially in preterm infants.

[Control of aspirin effect in chronic cardiovascular patients using two whole blood platelet function assays. PFA-100 and Multiplate]. / Vollblutaggregometrie zur Kontrolle der Wirksamkeit von Azetylsalizylsäure bei Patienten mit koronarer Herzkrankheit. Vergleich von PFA-100 und Multiplate.

In this study two aspirin sensitive platelet function tests which are based on the analysis of whole blood were evaluated and correlated with each other. In vitro bleeding time was determined using the PFA-100 analyzer (Dade Behring, Marburg, Germany) using the collagen/epinephrine cartridge and citrated blood. Whole blood aggregometry was performed using the Multiplate analyzer (Dynabyte medical, Munich, Germany) using hirudin blood (25 mug/ml). Aggregatin was triggered using arachidonic acid (ASPItest), collagen (COLtest) or TRAP-6 (thrombin receptor activating peptide, TRAPtest). Following informed consent citrated blood and hirudin blood was drawn from 76 cardiovascular patients which were on long-term aspirin therapy (aspirin patients). In addition hirudin blood was drawn from 57 healthy blood donors for assessment of whole blood aggregometry. PFA-100 closure times of the aspirin patients were 273 +/- 49 s. Based on the cut-off of 170 s a non response to the aspirin therapy was detected in 5 of 76 patients. Whole blood aggregation was comparable in the aspirin patients vs the blood donors AUC values in the TRAP test, whereas in COLtest and ASPItest significantly reduced aggregations were detected (p < 0.05). Of the five patients that had a normal PFA-100 closure time only one had normal aggregation in ASPItest, and also only one had a normal aggregation in COLtest. The high rate of response to the aspirin therapy which was found in PFA-100 and ASPItest can be explained by the assumed high level of compliance of the cohort. In the applied tests different patients were stratified as aspirin-non-responders. This highlights the importance of the assay conditions for the diagnosis of an aspirin-non-response.

[Merkel cell carcinoma--clinically often misjudged]. / Das Merkelzellkarzinom--klinisch oft verkannt.

Merkel cell carcinoma is a rare, rapidly growing, highly malignant dermal tumor which occurs preferentially on light-exposed skin in advanced age. The course of the disease is frequently characterized by the occurrence of lymph node metastases and local recurrences, even in the first year after removal of the primary tumour. The five-year overall survival rate is only about 65 %, despite rigorous therapy. The histological pattern is characterized by trabecular strands of small, uniform cells with large basophilic nuclei and typical neuroendocrine granules. The diagnosis is confirmed immunohistochemically by neuroendocrine and epithelial markers. The excision of the primary tumor is regarded as first-line therapy. Adjuvant radiatiotherapy is almost always indicated and should also include lymph node drainage. In the stage of nodal disease, a combination of excision and radiotherapy is recommended. Adjuvant chemotherapy can be applied in this stage, as in small-cell bronchial carcinoma. The prognostic advantage has, however, not been proven. Despite good response to radiatiotherapy and chemotherapy, with at least prolonged recurrence-free intervals, Merkel cell carcinoma is rarely curable at the distant metastasizing stage. Individually defined, aggressive treatment,including radiatiotherapy, may in future considerably improve the prognosis, especially in the early stages of the disease.

Multiple reversible MR signal changes caused by Epstein-Barr virus encephalitis.

This report presents the imaging findings of an unusual case of Epstein-Barr virus (EBV) encephalitis. A young man presented with a short-lasting history of febrile infection, neuropsychologic deficits, ataxia, and seizures. MR imaging revealed fully reversible signal intensities (T2, diffusion-weighted imaging with a decreased apparent diffusion coefficient) in the splenium of the corpus callosum and both posterior hemispheres. EBV infection must be added to the list of differential diagnoses of (reversible) splenial lesions.

Primary culture of microvascular endothelial cells from human benign prostatic hyperplasia.

BACKGROUND: Prostate growth seems to be influenced by paracrine factors like IL-6 originating from the microvascular endothelium. Therefore, our efforts were focused on the primary culture and behavior of microvascular endothelial cells (HPEC) derived from tissue of human benign prostatic hyperplasia (BPH). Until now, the isolation and culture of HPEC from BPH have not been reported. METHODS: BPH tissue was cut into small cubes and gently squeezed after incubation with dispase. HPEC were cultured from the resulting cell suspension after a stepwise selection by use of superparamagnetic beads coated with antibodies against endothelial specific antigens. HPEC were characterized by flow cytometry and immunohistochemistry. gamma-Glutamyl transpeptidase activity (specific for microvascular endothelium) was measured after dissolution of the HPEC with Triton X-100. After the incubation of HPEC either with ATP, VEGF, or TNF-alpha, the release of IL-6 was measured by enzyme linked immunosorbent assay (ELISA). RESULTS: HPEC showed a typical endothelial morphology. They were positive for von Willebrand factor, CD31, CD62E (after stimulation with TNF-alpha), alpha-actin and were negative for fibroblastic antigens and PSA. Proliferation was stimulated by vascular endothelial growth factor (VEGF). gamma-Glutamyl transpeptidase activity in HPEC was 6.3 microIU/microg protein, whereas in human umbilical vein endothelial cells (HUVEC) no gamma-glutamyl transpeptidase activity was detectable. The IL-6 secretion of HPEC was stimulated by VEGF and TNF-alpha, but not by ATP and bradykinin. CONCLUSIONS: For the first time, the primary culture of microvascular endothelial cells from BPH tissue was successfully performed. Our results suggest that HPEC may be actively involved in prostate growth, due to the secretion of regulatory factors such as IL-6.

Prognostic significance of the presence of erythroblasts in blood after cardiothoracic surgery.

In patients suffering from a variety of severe diseases the detection of erythroblasts in peripheral blood is associated with poor prognosis. However, as yet the prognostic significance of erythroblasts in the blood of patients after cardiothoracic surgery has not been assessed. In a retrospective study we analyzed the database of 2074 patients, of whom 87 died in hospital during the postoperative period. All patients underwent cardiothoracic surgery using a heart-lung machine. Together with erythroblasts in blood, age, sex, body mass index, preoperative ejection fraction, smoking, diabetes mellitus, type of operation, emergency surgery, renal deficiency, pulmonary hypertension, and endocarditis were considered. The postoperative mortality of patients with erythroblasts in peripheral blood (n=57) was 45.6% (n=26), being significantly higher (p<0.001) than the mortality of patients without erythroblasts (3.0%). None of six patients with more than 2000 erythroblasts x 10(6)/l survived. The postoperative detection of erythroblasts is highly predictive of death, the odds ratio after adjustment for the other known prognostic factors being 7.2 (95% confidence interval 3.4-15.1). Erythroblasts were detected for the first time on average 11 +/- 2 days (median: 7 days; n=57) after surgery and 8 +/- 2 days (median: 6 days; n=26) before death. The detection of erythroblasts in blood after cardiothoracic surgery has a high prognostic significance in terms of in-hospital mortality, helping physicians to identify patients at high risk of death. This finding has to be confirmed by a prospective study with the use of a more sensitive and reliable technology and prospectively defined time intervals for counting blood cells.

In vitro modulation of steroid 5alpha-reductase activity by phospholipases in epithelium and stroma of human benign prostatic hyperplasia.

Membrane components, such as phospholipids, play an important role in the regulation of prostatic 5alpha-reductase activity. To describe in more detail the impact of such regulation on 5alpha-reductase activity, epithelial and stromal cell homogenates of human BPH were treated with phospholipases to specifically alter the structure of cellular phospholipid components. Phospholipase A(2) (PLA(2)) was used to alter the structure of the nonpolar, hydrophobic region of the membrane bilayer. Various types of phospholipase C (PLC) affect the polar, hydrophilic region of phospholipids. In epithelium and stroma, 5alpha-reductase activity was dose-dependently inhibited by PLA(2) and PLC type III. In epithelium and stroma, the mean IC(50) values of PLA(2) were 9.4 +/- 1.1 and 13.9 +/- 2.6 [U/mg protein +/- SEM], respectively. The mean IC(50) values of PLC type III in epithelium and stroma were 4.5 +/- 1.2 and 1.7 +/- 0.2 [U/mg protein +/- SEM], respectively. In epithelium as well as in stroma, 5alpha-reductase activity was more greatly inhibited by PLC type III than by PLA(2). Both in epithelium and stroma, PLA(2) significantly decreased the V(max) of 5alpha-reductase whereas its K(m) remained unaffected. A similar decrease in V(max) was found with PLC type III in epithelium and stroma. Furthermore, the K(m) of epithelial 5alpha-reductase increased significantly following the addition of PLC type III. The two phospholipases, with their specific substrate affinities and sites of hydrolysis, exhibited significantly different effects on 5alpha-reductase, indicating that 5alpha-reductase activity is not unspecifically affected by modification of the hydrophilic milieu. Rather, 5alpha-reductase activity is specifically modulated by various phospholipids and/or phospholipolysis mediated degradation products. These findings suggest that the structural composition of the lipid environment plays a fundamental role in the post-translational regulation of 5alpha-reductase activity in the epithelium and stroma of human BPH. Thus, changes in membrane phospholipid content seem to be instrumental in the expression of DHT-dependent processes.

Transfer of biological traces in cases of hanging and ligature strangulation.

In hanging and ligature strangulation, the noose mostly causes a mark or groove which is formed partly by compression of the skin and partly by abrasion with loss of the upper epidermal layers. The horny scales abraded from the neck may be transferred to the strangulation device or to the interposed textiles where they are sometimes visible at stereomicroscopic examination or even to the naked eye as silver-grey particles. The morphologic features of the epidermal transfer due to hanging and ligature strangulation is demonstrated by 14 case examples. The biological traces may be sufficient for comparative DNA typing by means of PCR-based methods. In 9 out of the 14 cases, genomic DNA typing was successful. Analysis of mtDNA succeeded in another two cases, although genomic DNA could not be detected. Beside the accumulation of solid epidermic particles the paper describes deposition of serous and fatty tissue fluid at the ligature (mainly adjacent to skin ridges).

DNA typing of human remains found in damp environments.

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

Is it possible to differentiate mtDNA by means of HVIII in samples that cannot be distinguished by sequencing the HVI and HVII regions?

The mitochondrial control region includes three so-called hypervariable (HV) regions, in which the polymorphic positions show a particularly high frequency. According to a population study of 200 unrelated individuals from Germany, HVI (positions 16,024-16,365, according to Anderson) showed 88 variable positions in a total length of 342 bp (26%) and HVII (positions 73-340) displayed 65 mutable sites in 268 bp (24%). HVIII (positions 438-574) exhibited a slightly lower variability, with 25 polymorphic sites within 137 bp (18%), but contrasted clearly with the background, which showed variability rates of only 7% (positions 16,366-16,569, 1-72) and 3% (positions 341-437), respectively. At present, the displacement (D)-loop database in Magdeburg comprises 904 sequences of the mitochondrial HVI region and HVII region from Germans, Austrians and Swiss. By means of this material, the extent to which the mtDNA sequences that do not differ in the HVI and HVII regions can be differentiated by additionally sequencing HVIII was investigated.

[Significance of mitochondrial DNA for forensic stain analysis, identification and forensic lineage determination]. / Zur Bedeutung der mitochondrialen DNA für die forensische Spurenkunde, Identifizierung und Abstammungsbegutachtung.

DNA testing using conventional STR systems may produce insufficient results, if the genomic DNA in the specimen is either highly degraded or the available quantity is very small (e.g. skin particles, hair shafts or ancient bones). In some of these cases the examination of mitochondrial DNA, which is present in considerably larger copy numbers in the cytoplasm, is more successful than that of nuclear DNA. Identification of unknown corpses by conventional DNA typing sometimes remains doubtful, if only samples from presumably distant relatives or putative brothers or sisters are available for comparison. Since mitochondrial DNA is generally transmitted in maternal lineages, its sequence pattern can be directly compared with those of other individuals and, in case of the same maternal lineage, corresponding sequence chromatograms are to be expected. In connection with nuclear DNA typing methods certain sequence motives may furnish clues to ethnic groups. The report presents three cases illustrating the application possibilities of mtDNA typing in forensic practice.

Prognostic significance of low serum cholesterol after cardiothoracic surgery.

BACKGROUND: The precise prognostic significance of critically low cholesterol concentrations in patients undergoing cardiothoracic surgery is unknown. METHODS: In a retrospective case-control study, we analyzed the database of 2074 patients, of whom 87 died postoperatively in hospital. All patients underwent cardiothoracic surgery using a heart-lung machine. Age, sex, body mass index, preoperative ejection fraction, smoking, diabetes mellitus, type of operation, emergency surgery, renal deficiency, pulmonary hypertension, and endocarditis were considered together with serum concentrations of cholesterol, C-reactive protein, alanine aminotransferase, and triglycerides. The statistics included sensitivity, specificity, predictive value, odds ratio, and the ROC curve. RESULTS: Cholesterol decreased sharply immediately after surgery in both the deceased and the survivors. In the deceased, the mean cholesterol concentration (+/- SE) remained rather low between days 4 and 7 after surgery [2.46 +/- 0.16 mmol/L (95 +/- 6 mg/dL)]. In the survivors at that time, the mean cholesterol concentration was significantly (P <0.001) higher [4.37 +/- 0.03 mmol/L (169 +/- 1 mg/dL)]. The positive predictive value of a critically low cholesterol concentration [<3.10 mmol/L (<120 mg/dL)] was 25.4%, increasing to 66.6% at a cutoff value of 1.55 mmol/L (60 mg/dL). The odds ratio under those circumstances was 15.5, and the area under curve (C-statistic) was 0.90. CONCLUSIONS: The cholesterol concentration between days 4 and 7 after cardiothoracic surgery possesses a high prognostic significance in terms of in-hospital mortality.

Mitochondrial heteroplasmy among maternally related individuals.

The second hypervariable segment of the human mtDNA control region contains a homopolymeric tract of cytidines between nucleotides (nt) 303 and 315, interrupted by a thymidine at position 310, according to the Cambridge reference sequence. By direct sequencing, some individuals show blurred sequence chromatograms in this region which are not caused by a sequencing artefact but by high levels of length heteroplasmy. With respect to this length heteroplasmy ten maternally related individuals and two unrelated probands were examined. The relative proportions of length variants in the homopolymeric tract in selected individuals were determined by cloning and sequencing of multiple independent clones. All ten family members examined were heteroplasmic while the proportions of each genotype varied widely in different individuals. The size of a possible mitochondrial bottleneck during embryonic development of the offspring is discussed with respect to the changes in mitochondrial haplotypes within mother-offspring pairs. Our data are consistent with both slow and rapid segregation of mtDNAs between the generations, which would implicate a tight as well as a wide bottleneck. Therefore, a common bottleneck size in all individuals from this lineage seems to be very unlikely.

3D norm data: the first step towards semiautomatic virtual craniofacial surgery.

When planning craniofacial surgical interventions, the ideal appearance of the patient is very important. The final appearance should be as close as possible to that which the patient would have if he/she were without defects. Our first step towards achieving this is to build a database containing sets of three-dimensional CT images that allows for comparison of the shape of a patient with defects to the typical shape of an age- and sex-matched "average" person without defects. We started to collect CT data from patients without pathologies and, in co-operation with two radiology institutes (in Mannheim and Heidelberg), over 100 CT data sets have now been collected and classified according to age and sex. It is necessary to choose an appropriate statistical method to calculate the norm data from the different data sets. Based on the statistical method, an age- and sex-matched "average" model of the anatomy will be created.

PCR typing of human semen stains after SEM-EDX examination.

There is an increasing demand to use scanning electron microscopy in the forensic analysis of biological samples. Such analyses are routinely used for the investigation of blood stains, seminal stains, diatoms, residues on wounds and residues and trace elements in gunshot powder. The same samples are sometimes also required for identification via DNA analysis, e.g. blood stains, seminal stains or epidermal cells. The ionising radiation provokes damage to DNA and also to membrane and protein structure. The question therefore arises whether the usual sequence of such an investigation, i.e. prior application of SEM and afterwards analysis of DNA, can affect the success rate of the DNA analysis. We have therefore experimentally exposed semen samples to a defined electron beam for different time intervals varying between 1 and 25 min and afterwards performed quantitative and qualitative DNA analysis. Our studies revealed that sample treatment with an electron beam does not interfere with subsequent DNA typing by various currently used forensic PCR systems.

Allele frequencies for the VNTR locus D17S5 (YNZ22) in Hungary.

A population genetic study for the locus D17S5 was carried out on Caucasoids from the Budapest area consisting of 209 unrelated individuals. In this system we identified 13 different alleles and 51 genotypes. No new alleles were found and no significant deviation from Hardy-Weinberg equilibrium was observed. The power of discrimination was estimated at 0.958, the power of exclusion at 0. 60 and the observed heterozygosity at 0.80.

In vitro inhibition of androstenedione 5alpha-reduction by finasteride in epithelium and stroma of human benign prostatic hyperplasia.

Finasteride is a well known steroid 5alpha-reductase inhibitor. In this context, recently we have shown that in human benign prostatic hyperplasia (BPH) finasteride inhibits the 5alpha-reduction of testosterone to dihydrostestosterone (DHT) more effectively in the epithelium as compared to the stroma. The aim of the present study was to describe in epithelium and stroma of human BPH the effect of finasteride on the 5alpha-reduction of androstenedione, that is the second main circulating androgen in men, to androstanedione. Using a finasteride concentration of 75 nM and an androstenedione concentration of 220 nM, the mean inhibition [% +/- SEM] of 5alpha-reductase activity was significantly higher in epithelium (69 +/- 2) than in stroma (52 +/- 4). Both in epithelium and stroma, this inhibition of 5alpha-reductase activity was dose-dependent and competitive. Dixon plots as well as slope replots of Lineweaver-Burk plots showed that the mean inhibition constant Ki (nM +/- SEM) was significantly lower in epithelium (10 +/- 1 and 11 +/- 2, respectively) than in stroma (33 +/- 7 and 28 +/- 4, respectively) indicating a significantly stronger inhibitory effect of finasteride in epithelium. From those mean Ki values, it follows that in human BPH finasteride inhibits equally well both the 5alpha-reduction of androstenedione to androstanedione and testosterone to DHT. Based on these inhibition studies, there is no evidence for the coexistence of substrate-specific 5alpha-reductases converting either testosterone or androstenedione. However, the striking difference in finasteride sensitivity of the 5alpha-reduction between epithelium and stroma could be due to a cell-type specific expression of structurally different 5alpha-reductases as well as to a different access of finasteride to 5alpha-reductase in epithelium and stroma where, compared to each other, the lipid environment is significantly different.

Fatty acid composition of phospholipids in epithelium and stroma of human benign prostatic hyperplasia.

BACKGROUND: Although it is well known that prostatic 5alpha-reductase is active only in its membrane-bound form, rather limited information is available concerning the composition of cellular lipids in human BPH. Therefore, in the present study, the phospholipid fatty acid composition and content in epithelium and stroma of human BPH have been investigated for the first time. METHODS: Phospholipids separated on TLC plates were methylated and fatty acid methyl esters were analyzed by capillary gas chromatography. RESULTS: The fatty acid composition of total phospholipids was significantly different between epithelium and stroma. In particular, the percentage of oleic acid was significantly higher in epithelium as compared with stroma, whereas that of arachidonic acid was significantly lower in epithelium than in stroma. In addition, significant differences between epithelium and stroma were found in regard to the fatty acid composition of the main phospholipid subclasses. Another remarkable finding were the age-dependent changes of the fatty acid composition in human BPH. CONCLUSIONS: This study shows that the fatty acid composition of phospholipids is significantly different between epithelium and stroma of human BPH. Furthermore, age-dependent alterations of the fatty acid composition were found. Further studies are needed to determine whether the endogenous hormonal milieu in the prostate modulates the fatty acid composition of the prostatic cells, as well as what impact such modulation could have on the properties of membrane proteins, i.e., enzymes like the 5alpha-reductase and receptors, which are thought to be affected by alterations in membrane fluidity or composition, or both.

Location and frequency of polymorphic positions in the mtDNA control region of individuals from Germany.

In order to identify polymorphic positions and to determine their frequency in the human mitochondrial D-loop containing region, the mitochondrial DNA (mtDNA) control region of 200 unrelated individuals from Germany were amplified and directly sequenced. Sequence comparison led to the identification of 190 mitochondrial lineages as defined by 202 variable positions. The most frequently occurring lineage comprised 5 individuals, whereas 186 types of D-loop sequences were observed in only one individual. Of the sequences studied 7% are not unique but show at least one counterpart with an identical haplotype. The majority (61%) of the control regions investigated showed between four and eight nucleotide positions deviating from the reference sequence. The maximum number of deviations observed in a single control region was 18. The majority of the variable positions in the D-loop region (88%) are located within three hypervariable regions. Sequence variations are caused by nucleotide substitutions, insertions or deletions. As compared to insertions and deletions, nucleotide substitutions make up the vast majority of the mutations (90%). We have predominantly found transitions (75%) and a significantly lower frequency of transversions (15%) whereas insertions (6%) as well as deletions (4%) are rather rare. Upon sequencing the mitochondrial control region from 200 German Caucasians the genetic diversity was estimated at 0.99. The probability of two randomly selected individuals from a population having identical mtDNA types is 0.6%.