Comprehensive cancer predisposition testing within the prospective MASTER trial identifies hereditary cancer patients and supports treatment decisions for rare cancers.

BACKGROUND: Germline variant evaluation in precision oncology opens new paths toward the identification of patients with genetic tumor risk syndromes and the exploration of therapeutic relevance. Here, we present the results of germline variant analysis and their clinical implications in a precision oncology study for patients with predominantly rare cancers. PATIENTS AND METHODS: Matched tumor and control genome/exome and RNA sequencing was carried out for 1485 patients with rare cancers (79%) and/or young adults (77% younger than 51 years) in the National Center for Tumor Diseases/German Cancer Consortium (NCT/DKTK) Molecularly Aided Stratification for Tumor Eradication Research (MASTER) trial, a German multicenter, prospective, observational precision oncology study. Clinical and therapeutic relevance of prospective pathogenic germline variant (PGV) evaluation was analyzed and compared to other precision oncology studies. RESULTS: Ten percent of patients (n = 157) harbored PGVs in 35 genes associated with autosomal dominant cancer predisposition, whereof up to 75% were unknown before study participation. Another 5% of patients (n = 75) were heterozygous carriers for recessive genetic tumor risk syndromes. Particularly, high PGV yields were found in patients with gastrointestinal stromal tumors (GISTs) (28%, n = 11/40), and more specifically in wild-type GISTs (50%, n = 10/20), leiomyosarcomas (21%, n = 19/89), and hepatopancreaticobiliary cancers (16%, n = 16/97). Forty-five percent of PGVs (n = 100/221) supported treatment recommendations, and its implementation led to a clinical benefit in 40% of patients (n = 10/25). A comparison of different precision oncology studies revealed variable PGV yields and considerable differences in germline variant analysis workflows. We therefore propose a detailed workflow for germline variant evaluation. CONCLUSIONS: Genetic germline testing in patients with rare cancers can identify the very first patient in a hereditary cancer family and can lead to clinical benefit in a broad range of entities. Its routine implementation in precision oncology accompanied by the harmonization of germline variant evaluation workflows will increase clinical benefit and boost research.

Position sensitive measurement of trace lithium in the brain with NIK (neutron-induced coincidence method) in suicide.

Mood disorder is the leading intrinsic risk factor for suicidal ideation. Questioning any potency of mood-stabilizers, the monovalent cation lithium still holds the throne in medical psychiatric treatment. Furthermore, lithium`s anti-aggressive and suicide-preventive capacity in clinical practice is well established. But little is still known about trace lithium distribution and any associated metabolic effects in the human body. We applied a new technique (neutron-induced coincidence method "NIK") utilizing the 6Li(n,α)3H reaction for the position sensitive, 3D spatially resolved detection of lithium traces in post-mortem human brain tissue in suicide versus control. NIK allowed, for the first time in lithium research, to collect a three dimensional high resolution map of the regional trace lithium content in the non lithium-medicated human brain. The results show an anisotropic distribution of lithium, thus indicating a homeostatic regulation under physiological conditions as a remarkable link to essentiality. In contrast to suicide we could empirically prove significantly higher endogenous lithium concentrations in white compared to gray matter as a general trend in non-suicidal individuals and lower lithium concentrations in emotion-modulating regions in suicide.

Synthesis of controlled-size silver nanoparticles for the administration of methotrexate drug and its activity in colon and lung cancer cells.

A controlled synthesis of methotrexate (MTX) silver nanoparticles (AgNPs-MTX) using borohydride and citrate as reduction and reduction/capping agents, respectively, was performed in order to obtain AgNPs-MTX conjugates with a narrow size distribution. Their characterization showed polydispersed spherical shape nanoparticles with a mean size around 13 nm and distribution range between 7-21 nm. The presence of MTX was confirmed by FTIR and EDX analysis. Spectroscopic determinations suggest the chemisorption of MTX through a carboxylic group (-COOH) onto AgNPs via the exchange with a citrate molecule. Drug loading capacities calculated for AgNPs synthesized using different amounts of MTX were 28, 31 and 40%. In vitro drug release tests depicted similar release profiles for all conjugated amounts releasing between 77 to 85% of the initial MTX loaded into the AgNPs. With respect to free MTX, the addition of the nanocarrier delayed its release and also changed its pharmacokinetics. Free MTX is released after 3 hours following a first order kinetic model, whereas in the presence of AgNPs, a fast initial release is observed during the first 5 hours, followed by a plateau after 24 hours. In this case, AgNPs-MTX fitted a Higuchi model, where its solubilization is controlled by a diffusion process. Results obtained from flow cytometry of different cell lines treated with AgNPs-MTX demonstrated the combined anticancer effect of both reagents, decreasing the percentage of living cells in a colon cancer cell line (HTC-116) down to 40% after 48 hours of exposure. This effect was weaker but still significant for a lung cancer cell line (A-549). Finally, a zebrafish assay with AgNPs-MTX did not show any significant cytotoxic effect, confirming thereby the reduction of systemic drug toxicity achieved by coupling MTX to AgNPs. This observed toxicity reduction in the zebrafish model implies also a probable improvement of the usage of AgNPs-MTX in chemotherapy against human cancers.

[Publication performances of university clinics for anesthesiology: Germany, Austria and Switzerland from 2001 to 2010]. / Publikationsleistungen der Universitätskliniken für Anästhesiologie: Deutschland, Österreich und Schweiz von 2001-2010.

AIM: This study assessed the publication performance of university departments of anesthesiology in Austria, Germany and Switzerland. The number of publications, original articles, impact factors and citations were evaluated. MATERIAL AND METHODS: A search was performed in PubMed to identify publications related to anesthesiology from 2001 to 2010. All articles from anesthesiology journals listed in the fields of anesthesia/pain therapy, critical care and emergency medicine by the "journal citation report 2013" in Thomson Reuters ISI web of knowledge were included. Articles from non-anaesthesiology journals, where the stem of the word anesthesia (anes*, anaes*, anäst*, anast*) appears in the affiliation field of PubMed, were included as well. The time periods 2001-2005 and 2006-2010 were compared. Articles were allocated to university departments in Austria, Germany and Switzerland via the affiliation field. RESULTS: A total of 45 university departments in Austria, Germany and Switzerland and 125,979 publications from 2,863 journals (65 anesthesiology journals, 2,798 non-anesthesiology journals) were analyzed. Of the publications 23 % could not be allocated to a given university department of anesthesiology. In the observation period the university department of anesthesiology in Berlin achieved most publications (n = 479) and impact points (1,384), whereas Vienna accumulated most original articles (n = 156). Austria had the most publications per million inhabitants in 2006-2010 (n=50) followed by Switzerland (n=49) and Germany (n=35). The number of publications during the observation period decreased in Germany (0.5 %), Austria (7 %) and Switzerland (8 %). Tables 2 and 4-8 of this article are available at Springer Link under Supplemental. CONCLUSIONS: The research performance varied among the university departments of anesthesiology in Germany, Austria and Switzerland whereby larger university departments, such as Berlin or Vienna published most. Publication output in Germany, Austria and Switzerland has decreased. Data processing in PubMed should be improved.

Delay in the administration of appropriate antimicrobial therapy in Staphylococcus aureus bloodstream infection: a prospective multicenter hospital-based cohort study.

OBJECTIVES: Early broad-spectrum antimicrobial treatment reduces mortality in patients with septic shock. In a multicenter, prospective observational study, we explored whether delayed appropriate antimicrobial therapy (AAT) influences outcome in Staphylococcus aureus bloodstream infection (SAB). METHODS: Two hundred and fifty-six patients with SAB from ten German study centers were enrolled and followed for 3 months. Predisposing factors, clinical features, diagnostic procedures, antimicrobial therapy, and outcome were recorded. The appropriateness of antimicrobial therapy was judged by a trained physician based on in vitro activity, dosage, and duration of therapy. Therapy was considered to be delayed when more than 24 h elapsed between the first positive blood culture and the start of appropriate therapy. The association of delayed therapy with overall mortality and SAB-related events (i.e., attributable mortality or late SAB-related complications) was assessed by crosstabulation and propensity score-based logistic regression. RESULTS: One hundred and sixty-eight patients received AAT during their hospital stay, of whom 42 (25%) received delayed AAT. The overall mortality and the occurrence of severe sepsis or septic shock were lower in patients with delayed AAT, pointing towards confounding by indication. Adjusted 90-day mortality (adjusted odds ratio [OR] 0.91, 95% confidence interval [CI] [0.39-2.13], p 0.82) and SAB-related events (adjusted OR 1.46, 95% CI [0.47-4.51], p 0.52) also failed to show a significant impact of delayed AAT on outcome. CONCLUSION: In patients with SAB, early AAT may not improve survival. However, confounding by indication is a major challenge when analyzing and interpreting observational studies on the impact of delayed AAT.

Genotypic interaction and gender specificity of common genetic variants in the p53/mdm2 network in Crohn's disease.

BACKGROUND/AIMS: Defective p53-mediated apoptosis and cell cycle control have been implicated in the immunopathogenesis of Crohn's disease (CD). Since common functional variants of p53 (SNP72 G/C) and its key negative regulator mdm2 (SNP309 T/G) have been reported to affect cellular apoptotic and cell cycle arrest capacities, we assessed the effects of these variants on CD susceptibility and their relationship to NOD2/CARD15 as a well-established genetic CD risk factor. METHODS: The variants SNP72 G/C and SNP309 T/G were genotyped in 149 European CD patients and 478 healthy controls. Subgroup analysis was performed in relation to NOD2/CARD15 status and to demographic/clinical characteristics. RESULTS: The p53 SNP72 CC genotype tended to be less frequent in CD. This reached statistical significance only in the male cohort (0 vs. 7.3%; p = 0.037). Genotype and allele frequencies of both single-nucleotide polymorphisms (SNPs) were otherwise not significantly different. In the combined genotypic analysis, the genotype p53 SNP72 CC was significantly underrepresented in mdm2 SNP309 TT homozygotes (0 vs. 9.7%; p = 0.034). No association was observed between NOD2/CARD15 and the respective SNPs. CONCLUSION: We report on a gender-specific protective effect of the low-apoptotic SNP72 CC genotype, and a gender-unrestricted genotypic interaction between SNP309 TT and SNP72 CC, which, for the first time, links sequence variation of the p53/mdm2 network to CD, independent of NOD2/CARD15.

Microsatellite analysis in rheumatoid arthritis synovial fibroblasts.

OBJECTIVES: Rheumatoid arthritis (RA) is a chronic disease characterised by irreversible destruction of the affected joints. As aggressive transformed-appearing synovial fibroblasts are commonly found at the site of invasion of the rheumatoid synovium into the adjacent cartilage and bone, the presence of microsatellite instability (MSI) and expression of mismatch repair enzymes as a possible mechanism in the alteration of these cells was examined. METHODS: DNA was extracted from the synovial fibroblasts and blood of 20 patients with long term RA undergoing joint replacement, and the presence of MSI was studied at 10 microsatellite loci. In addition, immunohistochemistry was performed to evaluate the expression of the two major mismatch repair enzymes (hMLH1 and hMSH2) in rheumatoid synovium. RESULTS: MSI could not be detected in any of the fibroblast cell populations derived from the 20 different rheumatoid synovial samples. In addition, strong expression of mismatch repair enzymes could be seen in numerous cells, including fibroblasts, throughout the synovium. CONCLUSIONS: Applying the currently used and established markers for MSI, the data show for the first time that MSI does not appear to have an important role in alteration of rheumatoid synovial fibroblasts into an aggressive phenotype. On the other hand, strong mismatch repair enzyme synthesis in rheumatoid synovium supports the hypothesis of continuing DNA repair, presumably due to long term, inflammation induced DNA damage.

Preoperative staging and early postoperative complications in radical prostatectomy. Experiences in 35 cases.

It is already a textbook item that in patients with prostatic cancer stage T1-T2 N0M0 radical prostatectomy is the only curative treatment. Radical prostatectomy is indicated also for patients in stage T3 N0M0 who underwent antiandrogenic (Fugerel) treatment for 3 months with the aim of reducing tumour volume. In the following 35 cases will be scrutinized, with special regard to preoperative staging and early postoperative complications.

Temporal Fourier transform of digital angiograms for left ventricular regional wall motion analysis.

To determine whether or not the first harmonic of a temporal Fourier transform, applied pixel-by-pixel on time-intensity curves, can detect the subtle wall motion abnormalities due to ischemia, 6 dogs were instrumented with a micromanometer in the left ventricles, a hydraulic cuff occluder around the circumflex coronary artery, and sonomicrometers on the inferior (ischemic) and anterior (non-ischemic) walls. Left ventricular images, obtained after contrast injection via the pulmonary artery, were compared with dimension signals in control and 3 progressive levels of coronary stenosis (Stenosis I, II and III). Normalized, digital functional images (512 x 512 matrix, 256 shades of gray/pixel) were divided into anterior, apical, and inferior areas to acquire regional mean phase (degrees) and amplitude (intensity units) values. After inducing stenosis, phase in ischemic region significantly increased at all 3 levels of stenosis, whereas amplitude significantly decreased at Stenosis II and III. However, amplitude images showed clearly the topographic site of ischemia. There was a progressive increase in phase and decrease in amplitude in ischemic areas as the percent wall thickening (%WTh) fell (phase vs. %WTh: r = -0.55, p less than 0.005; amplitude vs. %WTh: r = 0.71, p less than 0.001). Heart rate and peak systolic pressure showed no significant changes during stenosis. We conclude that quantitative functional images, generated from a temporal Fourier transform, are sensitive to the detection of left ventricular regional wall motion abnormalities during mild, moderate, and severe degrees of ischemia.

Quantitative assessment of regional left ventricular function by densitometric analysis of digital-subtraction ventriculograms: correlation with myocardial systolic shortening in dogs.

Conventional wall motion analysis of contrast ventriculograms assesses only that part of the wall that is tangential to the x-ray beam. To assess regional left ventricular function in three dimensions, a new computerized method based on densitometric analysis of digital subtraction left ventriculograms was developed and validated in nine open-chest dogs instrumented with a circumflex coronary artery occluder and sonomicrometers in the anterior and posterior walls. Each dog underwent digital subtraction ventriculography at baseline and at five levels (I to V) of dysfunction of the inferior wall induced by progressive stenoses of the circumflex coronary artery. The ventriculogram was divided into six segments around the end-diastolic center of gravity. Time-volume curves were obtained by densitometry in the normal anterior and ischemic inferior segments containing the sonomicrometers. From these curves, regional ejection fraction (R-EF), regional peak ejection rate (R-PER), and regional phase (R-PH) and amplitude (R-AMP) of the first Fourier harmonic were derived. From baseline to level V of dysfunction, myocardial systolic shortening determined by sonomicrometry decreased by 124 +/- 34% of control (mean +/- SD; p less than .001) in the ischemic wall, while it increased by 12 +/- 19% (NS) in the normal wall. At the same time, R-EF, R-PER, and R-AMP decreased in the ischemic segment by 65 +/- 12%, 46 +/- 30%, and 45 +/- 15% of control, respectively (all p less than .01), while they remained unchanged or increased in the normal segment. R-PH was delayed by 14 +/- 5% (p less than .01) in the ischemic segment, but remained unchanged in the normal segment, reflecting the asynchrony of regional left ventricular contraction during ischemia. Densitometric indexes of regional function correlated well with sonomicrometric systolic shortening both in normal and ischemic segments, with r values of .84 for R-EF, .80 for R-AMP, .64 for R-PER, and .55 for R-PH (all p less than .0001). Thus, densitometric analysis of digital subtraction left ventriculograms allows three-dimensional assessment of the extent, velocity, and synchrony of regional left ventricular contraction. Densitometric indexes of regional contraction correlate well with direct measurements of myocardial systolic shortening and are useful in quantitating regional left ventricular dysfunction.

Decreased regional contractility in nonischemic myocardium during acute coronary artery occlusion in conscious pigs.

The purpose of this study was to use the relationship between end-systolic left ventricular pressure and segment length to assess the inotropic state of nonischemic myocardium during acute coronary artery occlusion in the conscious pig. Eight pigs were chronically instrumented with sonomicrometers to measure midwall segmental shortening and a micromanometer to measure left ventricular pressure. Occlusion of the inferior vena cava with a pneumatic occlusive cuff caused transient decreases in left ventricular pressure so that the relationship of left ventricular pressure and segment length at end systole could be determined over a range of pressures. In preliminary studies using open-chest pigs, this relation was shown to be highly linear and best quantified using a calculated segment length at a left ventricular pressure of 100 mm Hg (ESL100). During acute, 1-min occlusion of the left anterior descending coronary artery, the ESL100 of the nonischemic lateral and posterior walls was significantly increased from 8.75 +/- .18 mm to 9.64 +/- .21 mm (mean +/- SD, p less than .01), indicating a decreased inotropic state. Similarly, during occlusion of the left circumflex coronary artery, the ESL100 of the nonischemic anterior wall increased from 8.44 +/- 2.53 mm to 9.26 +/- 3.12 mm (p less than .05). This was not associated with a change in the amount of shortening during systole. Pharmacological autonomic blockade using atropine and propranolol failed to alter the response of nonischemic zones to acute coronary artery occlusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Densitometric regional ejection fraction: a new three-dimensional index of regional left ventricular function--comparison with geometric methods.

Densitometric regional ejection fraction obtained by computer analysis of digital subtraction ventriculography was evaluated as a new, quantitative, three-dimensional index of regional left ventricular performance. Eighteen patients with coronary artery disease and seven control subjects had right anterior oblique ventriculography at rest and immediately after rapid atrial pacing using central venous injection of contrast material. Regional left ventricular ejection fraction was determined by densitometry in six segments drawn around the end-diastolic center of gravity, and compared with two conventional indexes of segmental wall motion: area and radial regional ejection fraction. Densitometric, area or radial regional ejection fraction was classified as abnormal if it fell at least 2 standard deviations below the corresponding mean value in the normal group. The densitometric method did not require outlining of the end-systolic left ventricular silhouette and was the easiest and fastest to perform of all three techniques. In addition, intra- and interobserver reproducibilities were higher with the densitometric method (r = 0.97 and 0.95) than with either the area (r = 0.84 and 0.82) or the radial method (r = 0.82 and 0.76). Regional left ventricular dysfunction as assessed by the densitometric, area and radial techniques allowed the detection of coronary artery disease in 50, 50 and 44% of the patients at rest and in 83, 67 and 61% of the patients in the post-pacing period, respectively. Post-pacing regional left ventricular dysfunction accurately predicted the presence or absence of greater than 70% diameter stenosis in the supplying coronary artery in 75, 67 and 56% of the cases, respectively. Thus, densitometric analysis of digital subtraction ventriculography allows a fast and reproducible three-dimensional determination of regional left ventricular ejection fraction. Using this technique, pacing-induced regional dysfunction can be detected in most patients with coronary artery disease and corresponds well with the location of significant coronary artery lesions.

Application of end-systolic pressure-volume and pressure-wall thickness relations in conscious dogs.

The usefulness of end-systolic measures of ventricular function was compared with that of standard contractility indexes in conscious dogs. End-systolic relations between left ventricular pressure and volume and between pressure and wall thickness were analyzed in dogs previously instrumented with ultrasonic crystals. Progressive angiotensin infusions were used to generate computer-averaged pressure-volume and pressure-wall thickness loops. Both relations were linear in every study and highly reproducible. With low and high dose dobutamine, the end-systolic pressure-volume relations were significantly displaced, with increased slope and inconsistent changes in intercept. This relation was more useful than the ejection fraction for detecting contractility increases at different afterloads, but it showed no advantage over maximal left ventricular dP/dt at all ranges of preload and afterload. The end-systolic pressure-volume relations were insensitive for detecting mild decreases in inotropic state produced by propranolol, and maximal dP/dt was superior for detecting such mild acutely reduced contractility. The end-systolic pressure-wall thickness relations showed displacement with dobutamine, although slope and intercept changes were not significant; these relations did not detect mild decreases in contractility produced by propranolol. It is concluded that the end-systolic pressure-volume relation and a simplified end-systolic measure using pressure and wall thickness provide sensitive, load-independent and reproducible approaches for defining acute increases in left ventricular contractility in conscious animals. Maximal dP/dt was equally effective for defining these increases in contractility and more sensitive for detecting slight acute decreases in contractility.

Ischemic myocardial dysfunction assessed by temporal Fourier transform of regional myocardial wall thickening.

A Fourier analysis including the first 20 harmonics was performed on sonomicrometric measurements of regional myocardial wall thickness in eight conscious dogs under control conditions and at four levels of ischemia produced by a hydraulic occluder on the left circumflex coronary artery. Systolic wall thickening was reduced from 26.47 +/- 6.20% (S.D.) (control) to 22.05 +/- 5.73% (mild stenosis), 17.00 +/- 5.86% (moderate stenosis), 11.46 +/- 3.56% (severe stenosis), and 3.69 +/- 2.57% (30-second occlusion), values significantly different from each other (p less than 0.01). The amplitude of the first harmonic decreased stepwise from 1.35 +/- 0.31 to 1.08 +/- 0.29 mm, 0.90 +/- 0.27 mm, 0.69 +/- 0.24 mm, and 0.43 +/- 0.12 mm, all significantly different from each other (p less than 0.05). These amplitude values correlated to percent systolic wall thickening (r = 0.894, p = 0.001). A phase shift of the first harmonic from 137 +/- 11 to 139 +/- 14 degrees, 150 +/- 15 degrees (p less than 0.05 vs control), 161 +/- 21 degrees (p less than 0.01 vs control), and 191 +/- 21 degrees (p less than 0.01 vs control and severe stenosis) correlated with the increase in time from end diastole to the point of maximum wall excursion (r = 0.662, p less than 0.001). These data indicate that the extent of ischemic regional myocardial hypokinesis can be adequately described by the amplitude of the first harmonic, and that the asynchrony of ventricular contraction and relaxation can be detected from the phase of the first harmonic.

Nosocomial urinary tract infections: etiology and prevention.

Nosocomial urinary tract infections arise mainly after transurethral diagnostic or therapeutic operations and are favored by the raised susceptibility of many hospitalized patients to infection. Besides strict establishment of the indication, attention must be paid above all to proper performance of the catheterization. Standardization disposable sets are helpful in this regard. Choice of the suitable catheter type depends on the requirements of the application, the duration of drainage and on the anatomical conditions. Use of a closed urine drainage system must be part of the clinical standard today as well as mastery of daily catheter hygiene. Under observation of the contraindications, advantage should be taken of suprapubic catheter drainage.

Familial tumor of the testis.

A survey of the literature since 1928 yields reports on 100 patients with testicular tumor occurring in families. From our own case material we present clinical data on 11 additional patients. Non-twin brothers are most often involved. Presently available data are too limited to assess the impact of a possible genetic disposition on the development of malignoma of the testis. Additional case reports and register studies are needed. Equally undecided is the question if sperm banking of testicular tumor patients prior to therapy carries any genetic risk to the progeny.

[Sonography of the penis shaft]. / Sonographie des Penisschaftes.

The development of highly frequent transducers has created new diagnostic possibilities for the sonography in the area of the body of the penis. We controlled the valency of this technique on our patients. While the usual clinical and radiological methods still predominate in the diagnostics of traumas of the penis and diseases of the cavernous bodies, the ultrasound gives new possibilities of the exact classification in the carcinoma of the penis and the induratio penis plastica. Thus the basis for the development of alternative therapeutic methods is given.