[Detection of papillomavirus DNA in the prostate: a virus with underestimated clinical relevance?]. / Die Detektion von Papillomavirus-DNA in der Prostata : Ein Virus mit unterschätzter klinischer Relevanz?

BACKGROUND: Human papillomaviruses (HPV) are the most frequent pathogens of sexually transmitted diseases. They have been associated with an increased incidence of several anogenital tumors. Whether oncogenic HPV are involved in the pathogenesis of prostate cancer has been a subject of great controversy. This study's purpose was to investigate the association between HPV infection and prostate cancer (PCA). MATERIAL AND METHODS: The study included 213 consecutive patients with an average age of 65.7 (+/-8.4) years. Within the framework of transrectal, ultrasonic-guided multibiopsy of the prostate, one additional core was examined by means of polymerase chain reaction (PCR) in relation to bacterial, fungal, and viral (including HPV) DNA, with subsequent DNA sequencing. The collected data were correlated with the histological results and with diverse clinical variables. The influence of several predictors for the existence of PCA was verified with a logistic regression model. RESULTS: No general bacterial DNA (16S rDNA) was detected. Of the 213 patients, 145 (68.1%) showed HPV DNA. In 64% (n=137), high-risk HPV DNA were depicted; these were 18% of the total in each case of HPV genotypes 16 and 18. From our examinations, no significant positive correlation existed between the HPV evidence and the histologically verified PCA that was found in 23.5% of the patients (n=50; odds ratio 1.45; 95% confidence interval 0.71-2.91). The BK virus was not found in any of the cores confirmed through PCR. CONCLUSION: Although no positive correlation between HPV infection and PCA existed in our study, data from the literature suggest an influence of the papillomavirus on PCA oncogenesis. Future studies should highlight to what extent HPV DNA is inserted in the genome of prostate cells and is able to cause subsequent malignant transformation of particular genes.

Y-chromosome STR haplotypes in a population sample from Switzerland (Zurich area).

Allele frequencies and haplotypes for 12 Y-chromosome STR loci (DYS19, DYS385a/b, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438 and DYS439), included in the PowerPlex Y multiplex kit, were determined for a Swiss population sample of 150 male individuals. The gene diversities for the different loci were in the same range as shown for other European Population samples. The haplotype diversity was 0.9922. Pairwise haplotype analysis showed no significant differences in comparison with other European Population samples.

The impact of a register on the management of neonatal cooling in Switzerland.

BACKGROUND: Therapeutic hypothermia following hypoxic ischaemic encephalopathy in term infants was introduced into Switzerland in 2005. Initial documentation of perinatal and resuscitation details was poor and neuromonitoring insufficient. In 2011, a National Asphyxia and Cooling Register was introduced. AIMS: To compare management of cooled infants before and after introduction of the register concerning documentation, neuromonitoring, cooling methods and evaluation of temperature variability between cooling methods. STUDY DESIGN: Data of cooled infants before the register was in place (first time period: 2005-2010) and afterwards (second time period: 2011-2012) was collected with a case report form. RESULTS: 150 infants were cooled during the first time period and 97 during the second time period. Most infants were cooled passively or passively with gel packs during both time periods (82% in 2005-2010 vs 70% in 2011-2012), however more infants were cooled actively during the second time period (18% versus 30%). Overall there was a significant reduction in temperature variability (p < 0.001) comparing the two time periods. A significantly higher proportion of temperature measurements within target temperature range (72% versus 77%, p < 0.001), fewer temperature measurements above (24% versus 7%, p < 0.001) and more temperatures below target range (4% versus 16%, p < 0.001) were recorded during the second time period. Neuromonitoring improved after introduction of the cooling register. CONCLUSION: Management of infants with HIE improved since introducing the register. Temperature variability was reduced, more temperature measurements in the target range and fewer temperature measurements above target range were observed. Neuromonitoring has improved, however imaging should be performed more often.

Second primary glioblastoma.

Although characterized by a highly variable phenotype and multiple genetic alterations, glioblastomas are considered monoclonal in origin. We here report on a 64-yr-old patient who developed a second glioblastoma in the left frontal lobe 10 yr after surgical resection of a glioblastoma of right frontal lobe. The first tumor contained 2 p53 mutations, in codon 213 (CGA-->TGA, Arg-->stop) and codon 306 (CGA-->TGA, Arg-->stop), further, 1 missense PTEN mutation (codon 257, TTC-->TTA, Phe-->Leu) and a silent PTEN mutation (codon 154, TTC-->TTT, Phe-->Phe). The second glioblastoma also contained multiple, but different mutations: p53 mutations in codons 158 (CGC-->CAC, Arg-->His) and 273 (CGT-->TGT, Arg-->Cys), and a PTEN mutation in codon 233 (CGA-->TGA, Arg-->Stop). Both neoplasms had a homozygous p16 deletion. The discordant pattern of mutations indicates that the second glioblastoma was not a recurrence but an independent second glioblastoma. The presence in these neoplasms of multiple mutations in tumor suppressor genes suggests the involvement of a novel disease mechanism but there was no indication of a DNA mismatch repair deficiency or of an inherited tumor syndrome.

DNA fingerprinting: its application in forensic case work.

Forensic serology deals with cases of disputed paternity and criminal stains. The spectacular improvement using DNA-profiling is best demonstrated with cases of criminal stains where with the same ease as a suspect can be identified, an innocent person can be excluded. In cases with band-shifting, a statistical definition of a match applying Bayes theorem as a decision making tool seems mandatory since matching or non-matching cannot be treated as a binary event. Good postmortem DNA stability is found in brain cortex, lymph nodes and psoas muscle. DNA fingerprinting is also a perfect tool to investigate disputed identity of blood alcohol samples. In paternity cases, we recommend both multi locus and single locus probes in kinship cases, e.g. in mother-child-putative father cases as well as father-daughter and brother-sister incest cases, grandparent cases and two-men cases. For the biostatistical evaluation of SLP patterns the formal genetics for a system of multiple allelism is used and for multi locus probes the model of multiple diallelism is applicable.

Effects of selective oral antimicrobial prophylaxis and systemic antibiotics on the fecal flora and fecal beta-aspartylglycine concentration in patients with acute leukemia.

The influence of selective oral antimicrobial prophylaxis (SOAP) and systemic antibiotic treatment (SAT) on the intestinal flora of fourteen granulocytopenic patients was investigated. For SOAP, patients received Cotrimoxazole, Polymyxin and Nystatin; Gentamicin, Ticarcillin and Cefuroxime were administered for SAT. Under SOAP, a complete elimination of Enterobacteriaceae and a marked reduction of yeasts and staphylococci was found. A reduction in the number of anaerobic species was also observed; however, the concentration of anaerobic bacteria remained constant. In contrast, additional systemic antibiotic treatment caused a significant reduction of aerobic as well as anaerobic bacteria, except for microorganisms pathogenic to neutropenic patients (i.e. Staphylococcus epidermidis and Candida sp.). The fecal concentration of beta-aspartylglycine was inversely correlated with the presence of anaerobic bacteria.

Beta-endorphin: regional levels profile in the brain of the human infant.

Immunoradiometrical determinations of beta-endorphin (beta-EP) levels in 29 discrete brain regions from a series of victims of "Sudden Infant Death Syndrome" yielded a uniformly low levels profile in various areas of telencephalon, thalamus, pons, cerebellum and medulla oblongata. This low levels profile was interrupted by intermediate and high beta-EP levels in the midbrain and in two diencephalic zones. This study provides, for the first time, a comprehensive, neurochemically determined regional profile of beta-EP levels in the brain of the human infant.

Contamination of bronchoscopes with Mycobacterium tuberculosis and successful sterilization by low-temperature hydrogen peroxide plasma sterilization.

BACKGROUND: The transmission of mycobacteria by bronchoscopes has been reported several times in the last years. To explore methods to prevent transmission of tuberculosis in this way, we sterilized contaminated bronchoscopes with low-temperature hydrogen peroxide gas plasma sterilization. METHODS: Bronchoscopes were contaminated with Mycobacterium tuberculosis and decontaminated with a washer/disinfector ("normal washing"). Some were additionally disinfected with glutaraldehyde ("intensive washing"). Afterward the bronchoscopes were sterilized by low-temperature hydrogen peroxide plasma sterilization. RESULTS: After normal washing, 8/17 samples had positive results by culture, and 7/17 had positive results by nucleic acid amplification technique. After intensive washing, all samples had negative results by culture, and 10/25 had positive results by nucleic acid amplification technique; after sterilization with low-temperature hydrogen peroxide plasma sterilization, all samples had negative results by culture and nucleic acid amplification technique. CONCLUSION: Washing of bronchoscopes, as performed normally, is not sufficient for decontamination of bronchoscopes. Additional disinfection is recommended. If the nucleic acid amplification technique is used for diagnostic procedures, sterilization by low-temperature hydrogen peroxide plasma sterilization is recommended to avoid false-positive results.

Isolation and characterization of the flagellar hook of Campylobacter jejuni.

A method for purification of the flagellar hook of Campylobacter jejuni is described. The hook was shown to be composed of a subunit protein, which has a molecular mass of 92,000 and an isoelectric point of pI 4.8. A monoclonal antibody and a polyvalent antiserum was raised against the purified flagellar hook of C. jejuni. Immuno-electronmicroscopy revealed that the epitope recognized by the monoclonal antibody is surface-located. However, this antibody reacted only with the hook of the immunization strain, but not with other strains or other flagellated bacteria. Thus, our data indicate that the immunodominant epitopes are located on the surface of the hook and that these epitopes are strain-specific.

Role of murine macrophages and complement in experimental Campylobacter infection.

The roles of macrophages and the complement system as potential host defence mechanisms in mice against campylobacter infection were studied in vivo, by depleting the murine serum-complement or the phagocytic cells. Macrophage-depletion was performed by intraperitoneal (i.p.) injection of silica dust, Liquoid or dextran sulphate. During 5 days after infection, such mice showed a significant increase in mortality, compared with controls. In contrast, mice that were previously decomplemented by i.p. injection of Cobra Venom Factor showed no significant increase in mortality. The results with combined macrophage depletion and decomplementation did not differ from those with macrophage depletion alone. These experiments suggest that macrophages seem to be more important than complement in the defence of mice against experimental campylobacter infection.

A variable fluence step clustering and segmentation algorithm for step and shoot IMRT.

A step and shoot sequencer was developed that can be integrated into an IMRT optimization algorithm. The method uses non-uniform fluence steps and is adopted to the constraints of an MLC. It consists of a clustering, a smoothing and a segmentation routine. The performance of the algorithm is demonstrated for eight mathematical profiles of differing complexity and two optimized profiles of a clinical prostate case. The results in terms of stability, flexibility, speed and conformity fulfil the criteria for the integration into the optimization concept. The performance of the clustering routine is compared with another previously published one (Bortfeld et al 1994 Int. J. Radiat. Oncol. Biol. Ph.vs. 28 723-30) and yields slightly better results in terms of mean and maximum deviation between the optimized and the clustered protile. We discuss the specific attributes of the algorithm concerning its integration into the optimization concept.

Open regulation and practice in assisted dying.

The Netherlands, Oregon and Switzerland are the only areas in the world where assistance in dying has legally been practised in recent years. This article provides a detailed comparison of the history of the origins, legislation, monitoring systems and the extent of assistance in dying in these three places. It shows that the actual practice in Switzerland which, unlike Oregon, also allows assistance in suicide by means of infusions or gastric tubes, can today be technically quite similar to the permitted practice of active euthanasia on request in the Netherlands. Considering the preconditions restricting these practices, Swiss regulations are the most open, in that the law requires neither a medical second opinion (as in both the Netherlands and Oregon) nor the existence of a terminal illness (as in Oregon) as prerequisite to assistance in dying. In 2001, the proportion of assisted deaths (as reported to the authorities) in all deaths was almost ten times higher in the Netherlands (1.5% of all deaths) then in Oregon (<0.1% of all deaths) or Switzerland (0.2% of all deaths). The analysis of the different normative concepts underlying legislation reveals that in the Netherlands the basis for non-prosecution lies in the conflict of the physician's duties to respect life versus relief of suffering, while in the USA and in Switzerland the right-to-die concept plays a major role. These two concepts allow appreciation of distinctions between the roles of the physician in end-of-life practices and between assisted suicide and voluntary active euthanasia.

Agarose gel electrophoresis of human erythrocyte glutamic-pyruvic transaminase (GPT EC 2.6.1.2).

The method of choice to determine erythrocyte glutamate-pyruvic transaminase (GPT) including rare variants was starch gel electrophoresis. Methods using agarose as gel medium were not reported to our knowledge. We present an adapted method using the Tris/maleate buffer system and an agarose of low endosmosis. The common as well as rare variant types of GPT were quickly and reliably separated. In addition, a method for the consecutive determination of esterase D (ESD) and GPT on the same gel using the malic acid buffer system is described.

Postmortem stability of DNA.

High-molecular-weight DNA was recovered postmortem in sufficient quantities from various human organ tissues as well as from blood, although not all organs were equally well suitable. Good DNA stability was found in brain cortex, lymph nodes and psoas muscle over a period of three weeks postmortem. Spleen and kidney showed good DNA stability up to five days postmortem but after longer periods, rapid degradation was observed. Yields of DNA from blood were not consistent because of the non homogeneity of samples. Blood clots were rich with DNA. Generally, the amount of degraded DNA correlated directly with the duration of the postmortem period. However in some cases, DNA degradation was already prominent after a short period. However in some cases, DNA degradation was already prominent after a short period. Case histories showed that high environmental temperature at the site of death and/or infectious diseases prior to death were the main factors for rapid autolysis. Gradual disappearance to complete loss of the long fragments (15-23 kb) was observed in DNA fingerprinting using the minisatellite probe 33.15. No extra-bands were noted, thus excluding erroneous conclusions. However, evidentiary value of older samples was lower.

A report of an international collaborative experiment to demonstrate the uniformity obtainable using DNA profiling techniques.

This paper describes a collaborative exercise intended to demonstrate whether uniformity of DNA profile results could be achieved between different European laboratories. It was shown that this goal can be obtained provided that a common protocol is followed (specifically the use of a common electrophoretic buffer as being the most important parameter). Generally, lower molecular weight loci (with lower molecular weight fragments) such as YNH24 perform better than higher molecular weight loci such as MS43a. The results of the exercise are discussed in relation to the objectives of the European DNA profiling group (EDNAP).

Report on the second EDNAP collaborative STR exercise. European DNA Profiling Group.

The European DNA Profiling Group (EDNAP) has previously carried out collaborative exercises to determine which STR systems will produce results that can be reproduced by different laboratories. The first EDNAP exercise involving STR systems focused on different types of loci: a simple locus with six common alleles (HUMTH01) and a complex locus with > 35 alleles (ACTBP2). Generally the simpler STR system was found to be readily amenable for use across a wide range of different technologies, whereas a more complex locus presented difficulties. The second EDNAP STR exercise was intended to take the process of investigation a stage further. Some laboratories are developing automation, coupled with fluorescent methods of detection and multiplex applications, whereas others use manual methods involving visual detection techniques such as silver staining. The purpose of this exercise was to determine whether loci amenable to multiplexing with automation (as a quadruplex reaction) could also be successfully used with manual methods, either by multiplexing in duplex reactions or alternatively by using just a single pair of PCR primers.

Report of the European DNA profiling group (EDNAP)--towards standardisation of short tandem repeat (STR) loci.

This paper describes a collaborative exercise intended to demonstrate whether uniformity of DNA profiling results could be achieved between European laboratories using short tandem repeat (STR) loci. Two different STRs were chosen--HUMTH01 and the AT-rich HUMACTBP2 (SE33). The former locus has only five common alleles, whereas the latter is complex and has at least 30 alleles. Laboratories were asked to test seven blood stains and to report the results to the coordinating laboratory. The exercise demonstrated that the simple STR systems such as HUMTH01 are more amenable to adoption as standard loci than complex AT-rich systems.

Report of a European collaborative exercise comparing DNA typing results using a single locus VNTR probe.

A collaborative exercise was carried out in 1989 among 12 European forensic laboratories using the single locus VNTR probe pYNH24, the restriction enzyme HinfI, the same set of human genomic DNA samples, and a standardized DNA size marker. The objectives of the exercise were: (1) to study the degree of variation within and between laboratories, (2) to obtain information on requirements for technical standardization allowing the exchange of typing results and (3) to compare different approaches for the identification of allelic DNA fragments of unknown size. Each laboratory carried out up to 10 independent typing experiments using the same DNA samples. The results were analysed independently by two laboratories using three different methods. The results of the exercise demonstrate the correlation of typing that can be achieved within and between laboratories under conditions of minimal standardization.

[Species diagnosis of a tissue using the polymerase chain reaction]. / Speziesdiagnostik an einem Gewebe mit Hilfe der Polymerase-Ketten-Reaktion.

The unambiguous identification of a biological specimen can deliver invaluable evidence to solve criminal cases. In this case the origin of a heart had to be clarified. Using the polymerase chain reaction technique and species-specific primer pairs for two genes it was clearly shown that this tissue was not from a human but from a pig.

[Change in the drinking behavior in Central Switzerland over two decades]. / Zur Anderung des Trinkverhaltens in der Zentralregion der Schweiz während zweier Jahrzehnte.

Based on the medical recordings registered during blood sampling of drunken drivers of the years 1971 and 1989 the changes of drinking behaviour in Switzerland was examined. An increase of female drivers under the influence of alcohol was noted. The younger drivers under 30, which were the most represented in all the age groups, showed a decrease in the average blood alcohol value. Due to the facts, the most blood samples were taken by forensic physicians rather than private physicians a much better information was gained concerning the substances drunken and the times of drinking. The evaluation of final drinking hours, critical incidents and time of blood sampling showed a tendency to post-midnight hours.