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Purpose: Population-scale, exome-sequenced cohorts with linked electronic health records (EHR) permit genome-first exploration of phenotype. Phenotype and cancer risk are well characterized in children with a pathogenic DICER1 (HGNC ID:17098) variant. Here, the prevalence, penetrance, and phenotype of pathogenic germline DICER1 variants in adults were investigated in 2 population-scale cohorts. Methods: Variant pathogenicity was classified using published DICER1 ClinGen criteria in the UK Biobank (469,787 exomes; unrelated: 437,663) and Geisinger (170,503 exomes; unrelated: 109,789) cohorts. In the UK Biobank cohort, cancer diagnoses in the EHR, cancer, and death registry were queried. For the Geisinger cohort, the Geisinger Cancer Registry and EHR were queried. Results: In the UK Biobank, there were 46 unique pathogenic DICER1 variants in 57 individuals (1:8242; 95% CI: 1:6362-1:10,677). In Geisinger, there were 16 unique pathogenic DICER1 variants (including 1 microdeletion) in 21 individuals (1:8119; 95% CI: 1:5310-1:12,412). Cohorts were well powered to find larger effect sizes for common cancers. Cancers were not significantly enriched in DICER1 heterozygotes; however, there was a ∼4-fold increased risk for thyroid disease in both cohorts. There were multiple ICD10 codes enriched >2-fold in both cohorts. Conclusion: Estimates of pathogenic germline DICER1 prevalence, thyroid disease penetrance, and cancer phenotype from genomically ascertained adults are determined in 2 large cohorts.

The polarization observables T,E,P,H, and G in photoproduction of η mesons off protons are measured for photon energies from threshold to W=2400 MeV (T), 2280 MeV (E), 1620 MeV (P,H), or 1820 MeV (G), covering nearly the full solid angle. The data are compared to predictions from the SAID, MAID, JüBo, and BnGa partial-wave analyses. A refit within the BnGa approach including further data yields precise branching ratios for the Nη decay of nucleon resonances. A Nη-branching ratio of 0.33±0.04 for N(1650)1/2− is found, which reduces the large and controversially discussed Nη-branching ratio difference of the two lowest mass JP=1/2−-resonances significantly. Keywords: Baryon spectroscopy, Meson photoproduction, Polarization observables

We report a first measurement for ultrahigh energy cosmic rays of the correlation between the depth of shower maximum and the signal in the water Cherenkov stations of air-showers registered simultaneously by the fluorescence and the surface detectors of the Pierre Auger Observatory. Such a correlation measurement is a unique feature of a hybrid air-shower observatory with sensitivity to both the electromagnetic and muonic components. It allows an accurate determination of the spread of primary masses in the cosmic-ray flux. Up till now, constraints on the spread of primary masses have been dominated by systematic uncertainties. The present correlation measurement is not affected by systematics in the measurement of the depth of shower maximum or the signal in the water Cherenkov stations. The analysis relies on general characteristics of air showers and is thus robust also with respect to uncertainties in hadronic event generators. The observed correlation in the energy range around the ‘ankle’ at lg⁡(E/eV)=18.5–19.0 differs significantly from expectations for pure primary cosmic-ray compositions. A light composition made up of proton and helium only is equally inconsistent with observations. The data are explained well by a mixed composition including nuclei with mass A>4. Scenarios such as the proton dip model, with almost pure compositions, are thus disfavored as the sole explanation of the ultrahigh-energy cosmic-ray flux at Earth. Keywords: Pierre Auger Observatory, Cosmic rays, Mass composition, Ankle

The first measurements of the beam-target-helicity-asymmetries E and G in the photoproduction of ω-mesons off protons at the CBELSA/TAPS experiment are reported. E (G) was measured using circularly (linearly) polarised photons and a longitudinally polarised target. E was measured over the photon energy range from close to threshold (Eγ=1108 MeV) to Eγ=2300 MeV and G at a single energy interval of 1108

Data on the polarization observables T, P, and H for the reaction γp→pπ0 are reported. Compared to earlier data from other experiments, our data are more precise and extend the covered range in energy and angle substantially. The results were extracted from azimuthal asymmetries measured using a transversely polarized target and linearly polarized photons. The data were taken at the Bonn electron stretcher accelerator ELSA with the CBELSA/TAPS detector. Within the Bonn-Gatchina partial wave analysis, the new polarization data lead to a significant narrowing of the error band for the multipoles for neutral-pion photoproduction.

Background: Thoracoabdominal normothermic regional perfusion (TA-NRP) has emerged as a powerful technique for optimizing organ procurement from donation after circulatory death donors. Despite its rapid adoption, standardized guidelines for TA-NRP implementation are lacking, prompting the need for consensus recommendations to ensure safe and effective utilization of this technique., Methods: A working group composed of members from The American Society of Transplant Surgeons, The International Society of Heart and Lung Transplantation, The Society of Thoracic Surgeons, and The American Association for Thoracic Surgery was convened to develop technical guidelines for TA-NRP. The group systematically reviewed existing literature, consensus statements, and expert opinions to identify key areas requiring standardization, including predonation evaluation, intraoperative management, postdonation procedures, and future research directions., Results: The working group formulated recommendations encompassing donor evaluation and selection criteria, premortem testing and therapeutic interventions, communication protocols, and procedural guidelines for TA-NRP implementation. These recommendations aim to facilitate coordination among transplant teams, minimize variability in practice, and promote transparency and accountability throughout the TA-NRP process., Conclusions: The consensus guidelines presented herein serve as a comprehensive framework for the successful and ethical implementation of TA-NRP programs in organ procurement from donation after circulatory death donors. By providing standardized recommendations and addressing areas of uncertainty, these guidelines aim to enhance the quality, safety, and efficiency of TA-NRP procedures, ultimately contributing to improved outcomes for transplant recipients., Competing Interests: Disclosures The authors have no conflicts of interest to disclose., (Copyright © 2024 The Transplantation Society and by Elsevier, Inc on behalf of The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

Objectives: The developmental and epileptic encephalopathies (DEE) are a heterogeneous group of rare neurodevelopmental disorders, characterised by early-onset seizures that are often intractable, electroencephalographic abnormalities, and developmental delay or regression. There is a paucity of data from sub-Saharan Africa on the genetic basis of DEE. The aim of this study was to investigate the genetic background of DEE using targeted next generation sequencing (NGS) analysis in a tertiary pediatric neurology outpatient department at Tygerberg Hospital, South Africa. In addition, we assessed the value of the genetic results to the parents and managing physicians., Methods: A prospective cohort study of 41 consecutive children with DEE (onset before 3 years of age) that were recruited over a 2-year period (2019-2021). Pre- and post-test genetic counselling were offered to all study participants. The results were categorized as either: positive (pathogenic/likely pathogenic variant identified), inconclusive (variant(s) of unknown significance identified), or negative (no variants identified). Result interpretation and careful matching of the variant to the clinical phenotype was performed. Subsequently, questionnaires were administered to both the physicians and the parents., Results: A genetic underlying cause for DEE was identified in 18 of 41 children (diagnostic yield 43.9%). Variants in SCN1A (n=7), KANSL1 (n=2), KCNQ2 (n=2) and CDKL5 (n=2) were identified in more than one patient. Rarer genes included IQSEC2, SMC1A and STXBP1. All of the identified pathogenic variants fully explained and matched the respective phenotypic description of the patient at the time of clinical diagnosis. In 26% of patients the genetic result facilitated precision medicine management changes to anti-seizure medication. Both parents and physicians expressed benefit of genetic testing in patients with DEE., Conclusion: Targeted NGS analysis proved an efficient diagnostic tool in detection of a genetic cause of DEE in a large proportion of South African children. The 43.9% diagnostic yield is similar to previously reported international pediatric cohorts. Additionally, the genetic findings proved useful for targeted therapeutic decision-making and accurate genetic counseling., Competing Interests: Declaration of Competing Interests None of the authors has any conflict of interest to disclose. We confirm that we have read the Journal's position on issues involved in ethical publication and affirm that this report is consistent with those guidelines. The authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript., (Copyright © 2022 British Epilepsy Association. Published by Elsevier Ltd. All rights reserved.)

Donation after circulatory death is emerging as an alternative pathway to donation after brain death to expand the cardiac organ donor pool. We describe the surgical technique and circuit configuration for in-situ organ reperfusion with thoracoabdominal normothermic regional perfusion using portable venoarterial extracorporeal membrane oxygenation., (Copyright © 2022 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

This paper presents a unique approach to the Impact Evaluation of a project that focused on low-threshold intergenerational play-based interactions in order to support young children from marginalised communities in eight European countries. The approach builds upon the work of Fetterman's Empowerment Evaluation and Patton's Utilization Focused Evaluation and brings them together to form an adapted model of evaluation. We outline in this paper how these two well developed methods of evaluation have been applied to a real world context, that is, the impact evaluation of a complex international project. Our approach highlights the complexities of differing contexts and allows for surprising and unintended consequences to emerge. It results, through double loop learning, a type of feedback loop with the internal stakeholders and implementers that is useful to the project coordination team, with a view to further upscaling of the initiative. Recommendations for policy at local, national and European Union levels were provided to the project and potential external users. However, the predominant feedback was provided at two crucial points along the way; during a stakeholder mapping exercise and during the further development of monitoring data tools., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Lung transplantation with lungs procured from donors after circulatory death (DCD) has been established as an alternative technique to traditional donation after brain death (DBD) with comparable outcomes. Recently, in situ thoracoabdominal normothermic regional perfusion (TA-NRP) has emerged as a novel technique employed in the procurement of cardiac allografts after circulatory death. TA-NRP, in contrast to ex situ machine perfusion, has the advantage of allowing in situ assessment of donor organs prior to final acceptance. However, there are some concerns that this technique may adversely impact the quality of lung allografts. Here, we present a case of a successful bilateral sequential lung transplantation in a patient with postinflammatory pulmonary fibrosis due to acute respiratory distress syndrome (ARDS), with lungs procured after normothermic in situ lung perfusion. Apart from the lungs, heart, liver, and kidneys were also successfully transplanted from this donor., (© 2021 The American Society of Transplantation and the American Society of Transplant Surgeons.)

Gene expression signatures have proven their potential to characterize important cancer phenomena like oncogenic signaling pathway activities, cellular origins of tumors, or immune cell infiltration into tumor tissues. Large collections of expression signatures provide the basis for their application to data sets, but the applicability of each signature in a new experimental context must be reassessed. We apply a methodology that utilizes the previously developed concept of coherent expression of genes in signatures to identify translatable signatures before scoring their activity in single tumors. We present a web interface (www.rosettasx.com) that applies our methodology to expression data from the Cancer Cell Line Encyclopaedia and The Cancer Genome Atlas. Configurable heat maps visualize per-cancer signature scores for 293 hand-curated literature-derived gene sets representing a wide range of cancer-relevant transcriptional modules and phenomena. The platform allows users to complement heatmaps of signature scores with molecular information on SNVs, CNVs, gene expression, gene dependency, and protein abundance or to analyze own signatures. Clustered heatmaps and further plots to drill-down results support users in studying oncological processes in cancer subtypes, thereby providing a rich resource to explore how mechanisms of cancer interact with each other as demonstrated by exemplary analyses of 2 cancer types., (Copyright © 2021. Published by Elsevier Inc.)

The problems of determining the protective properties of barrier materials used for the production of body surface protection products of an isolative type are very actual. These garments are expected to have long-term resistance against permeation of toxic substances. The paper deals with the study of the effect of organic solvent vapours on the changes in protective properties of selected anti-gas protective garments commonly used by the North Atlantic Treaty Organization armed forces. Permeation measurements of selected gases with integral permeameter were performed to verify their barrier properties, surface changes after exposure to selected solvents by using a 3D optical profilometer, permeation measurements for vapours of these solvents with differential permeameter, and experimental simulating the possible influence of barrier properties of garments within permeation of air. It has been shown that in the case of gases, the permeability of the studied materials increases with decreasing kinetic diameter of the penetrant molecule used. In the case of acetonitrile and isooctane vapours, permeability increased with increasing vapour concentration due to interactions between polymeric materials and molecules of organic compounds. The permeation measurements of pure vapours and air/vapour mixtures indicate that the level of interactions between the molecules of the penetrant and the material of the protective garment is not strong enough to degrade the material of the garment in such a way as to allow a greater penetration of the pollutants through it. The results of permeation experiments for gases and organic vapours showed very good barrier properties of studied chemical isolated garments., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

We present a case of a young female patient with end-stage lung failure because of pulmonary arterial hypertension who was failing maximal medical therapy and was listed for a single sequential lung transplantation. The challenge of the case was a concomitant presence of a large atrial septal defect. The novelty of our approach was a device closure of atrial septal defect before performing transplantation with the use of intraoperative venoarterial extracorporeal membrane oxygenation., (Copyright © 2018 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

Background: Partially implantable active middle ear implants (aMEIs) offer a solution for individuals who have mild to severe sensorineural hearing loss and an outer ear medical condition that precludes the use of hearing aids. When otherwise left untreated, individuals report a lower quality of life, which may further decrease with increasing disability. In the lack of cost-effectiveness studies and long-term data, there is a need for decision modeling., Objective: To explore individual-level variance in resource utilization patterns following aMEI implantation., Methods: A Markov model was developed and analyzed as microsimulation to estimate the incremental cost utility ratio (ICUR) of partially implantable aMEIs compared with no (surgical) intervention in individuals with sensorineural hearing loss and an outer ear medical condition in Australia. Cost data were derived mostly from the Medicare Benefit Schedule and effectiveness data from published literature. A third-party payer perspective was adopted, and a 5% discount rate was applied over a 10-year time horizon., Results: Compared with baseline strategy, aMEIs yielded an incremental cost of Australian dollars (AUD) 13,339.18, incremental quality-adjusted life-year (QALY) of 1.35, and an ICUR of AUD 9,913.72/QALY. Of the respective number of simulated patients who visited each health state, 75.73% never had a minor adverse event, 99.82% did not experience device failure, and 97.75% did not cease to use their aMEIs. Probabilistic sensitivity analyses showed the ICUR to differ by only 0.95%., Conclusions: In the Australian setting, partially implantable aMEIs offer a safe and cost-effective solution compared with no intervention and are also well accepted by users., (Copyright © 2017 International Society for Pharmacoeconomics and Outcomes Research (ISPOR). Published by Elsevier Inc. All rights reserved.)

Background: This study examined whether aortic valve opening (AVO) and other echocardiographic parameters influence outcomes in patients on left ventricular (LV) assist device (LVAD) support. Pump thrombosis (PT) and ischemic stroke (IS) are known complications of LVAD, but mechanisms that could influence them are not completely understood., Methods: This was a retrospective analysis of 147 patients who received a HeartWare Ventricular Assist Device ( HeartWare International) as a bridge to transplant or to candidacy between July 2009 and August 2015, of whom 126 had at least 30 days of follow-up before the first event (30-days-out cohort). Outcomes included survival, PT, IS, and PT+IS (combined thrombotic event; CTE)., Results: Median time on support was 518 days. Of the 30-days-out cohort, 29% had a first PT and 19% a first IS. AVO was associated with longer survival on device (1,081 vs 723 days; p = 0.01) in the entire cohort. In the 30-days-out cohort, the aortic valve was more frequently closed in patients with lower ejection fractions on support (14% ± 6% vs 18% ± 9%; p = 0.009), more dilated pre-event echocardiogram (LV end-diastolic diameter, 66 ± 12 mm vs 62 ± 10 mm; p = 0.04), and pre-implant LV end-diastolic diameter (70 ± 10 mm vs 66 ± 9 mm; p = 0.06). CTE-free survival on the device was lower with a closed aortic valve (897 vs 1,314 days; p = 0.003) as was PT-free survival on the device (1,070 vs 1,457 days; p = 0.02). Cox regression analysis showed that AVO was an independent predictor of CTE (p = 0.03) CONCLUSIONS: Thrombotic events are relatively frequent in patients on long-term LVAD support. A closed aortic valve was associated with decreased overall survival, thrombosis-free survival, and poorer LV function on support. These are high-risk patients, so whether they require more intense anti-coagulation or prioritizing for transplantation requires further research., (Copyright © 2016 International Society for Heart and Lung Transplantation. Published by Elsevier Inc. All rights reserved.)

A 64-year-old woman with extensive tumorous infiltration of the mitral and aortic valves underwent partial resection of a tumor of the left ventricular outflow tract and replacement of both affected valves. Histology revealed an inflammatory pseudotumor with a significant number of immunoglobulin-G4-positive plasma cells. The histologic and clinical findings suggested immunoglobulin-G4-related disease of the heart., (Copyright © 2015 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

Background: The potential role of genitourinary infection in the etiology of prostate cancer (CaP) has been extensively investigated for 30 years. Two basic approaches have been used: tissue-based methods (polymerase chain reaction, immunohistochemistry, and in situ hybridization) and serologic assays (enzyme-linked immunosorbent assay, immunofluorescence, etc.). The objective of this review was to answer the question of whether infection of the male genitourinary tract may have a role in the etiology of CaP., Materials and Methods: We have carried out a systematic review of the evidence that was published in the MEDLINE/PubMed database until December 2011. The search terms included "prostate cancer," "infection," and the explicit names of the various infectious agents. Additional studies were identified using a reference search. A total of 74 papers were included in the review, which cover the following infectious agents: human papillomavirus, cytomegalovirus, herpes simplex virus, Epstein-Barr virus, human herpesvirus, BK virus, JC virus, chlamydia, mycoplasma, ureaplasma, trichomonas, neisseria, treponema, Propionibacterium acnes, xenotropic murine leukemia virus-related virus and Candida albicans., Results: Despite the variable study designs and methodological approaches that were used, most of the pathogens that were studied were unlikely to be directly involved in prostate carcinogenesis., Conclusions: The role of infection in the etiology of CaP has yet to be determined despite 30 years of research efforts. A discovery of an infectious agent that is associated with CaP would be of great medical importance; however, such a link would have to be firmly established before impacting on patient care., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Study Objective: To determine whether ultrasound guidance improves the quality of continuous popliteal block when compared with a nerve stimulator after major foot and ankle surgery., Design: Prospective, randomized, double-blinded clinical trial., Setting: Operating room, Postanesthesia Care Unit (PACU), and hospital wards of a university-affiliated hospital., Patients: 45 ASA physical status 1, 2, and 3 patients undergoing elective major foot and ankle surgery., Interventions: Placement of a popliteal sciactic nerve catheter using either nerve stimulator or ultrasound guidance. In the PACU, a continuous infusion of ropivacaine 0.2% was started at a basal rate of 4 mL/hr and adjusted in a standardized fashion to maintain visual analog scale (VAS) pain scores < 4. All patients also received intravenous (IV) patient-controlled analgesia with hydromorphone and oral opioids., Measurements: VAS pain scores at rest and with physical therapy, ropivacaine use, opioid use, and opioid-related side effects were recorded., Main Results: Cummulative ropivacaine use was lower in patients whose catheter was placed by ultrasound than by nerve stimulator guidance (mean 50 vs 197 mL, P < 0.001). Pain scores at rest and during activity were similar between groups. Cumulative opioid consumption (mean 858 vs 809 mg oral morphine equivalents) and daily frequencies of nausea (5% to 33% vs 0 to 24%) and pruritus (0 to 21% vs 0 to 24%) were similar between groups. Length of hospital stay was similar between groups (3.5 vs 3.7 days)., Conclusions: Ultrasound guidance was associated with less local anesthetic consumption than with the nerve stimulator; however, there was little clinical benefit, as all other outcomes were similar between groups., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background Aims: Mesenchymal stromal cells (MSC) are the focus of research in regenerative medicine aiming at the regulatory approval of these cells for specific indications. To cope with the regulatory requirements for somatic cell therapy, novel approaches that do not interfere with the natural behavior of the cells are necessary. In this context in vivo magnetic resonance imaging (MRI) of labeled MSC could be an appropriate tool. Cell labeling for MRI with a variety of different iron oxide preparations is frequently published. However, most publications lack a comprehensive assessment of the non-interference of the contrast agent with the functionality of the labeled MSC, which is a prerequisite for the validity of cell-tracking via MRI., Methods: We studied the effects of iron oxide-poly(l-lactide) nanoparticles in MSC with flow cytometry, transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), Prussian blue staining, CyQuant® proliferation testing, colony-forming unit-fibroblast (CFU-F) assays, flow chamber adhesion testing, immunologic tests and differentiation tests. Furthermore iron-labeled MSC were studied by MRI in agarose phantoms and Wistar rats., Results: It could be demonstrated that MSC show rapid uptake of nanoparticles and long-lasting intracellular persistence in the endosomal compartment. Labeling of the MSC with these particles has no influence on viability, differentiation, clonogenicity, proliferation, adhesion, phenotype and immunosuppressive properties. They show excellent MRI properties in agarose phantoms and after subcutaneous implantation in rats over several weeks., Conclusions: These particles qualify for studying MSC homing and trafficking via MRI.

Fibrodysplasia ossificans progressiva (FOP) is a severe genetic disorder in which progressive ossification of subcutaneous tissues leads to immobility and profound physical handicap. Dental management of affected persons may be constrained by impaired mobility of the temporomandibular joints. Equally, the traumatic aspects of dental intervention can exacerbate the ossification process. In the later stages of the disorder, thoracic immobility impairs pulmonary ventilation and compromises dental anesthesia. We have undertaken dental appraisal and treatment of 5 South Africans with FOP, with ages ranging from 2 to 52 years. The age relationship of the manifestations of FOP was apparent in these persons, as were the dental problems that emerged. These dental observations illustrate the importance of accurate diagnosis and the awareness of potential complications that may be encountered in dental management. Our observations are presented and discussed in this article., (Copyright © 2011 Mosby, Inc. All rights reserved.)

Current quantitative metabolomic research in brain tissue is challenged by several analytical issues. To compare data of metabolite pattern, ratios of individual metabolite concentrations and composed classifiers characterizing a distinct state, standardized workup conditions, and extraction medium are crucial. Differences in physicochemical properties of individual compounds and compound classes such as polarity determine extraction yields and, thus, ratios of compounds with varying properties. Also, variations in suppressive effects related to coextracted matrix components affect standards or references and their concentration-dependent responses.The selection of a common tissue extraction protocol is an ill-posed problem because it can be regarded as a multiple objective decision depending on factors such as sample handling practicability, measurement precision, control of matrix effects, and relevance of the chemical assay. This study systematically evaluates the impact of extraction solvents and the impact of the complex brain tissue on measured metabolite levels, taking into account ionization efficiency as well as challenges encountered in the trace-level quantification of the analytes in brain matrices. In comparison with previous studies that relied on nontargeted platforms, consequently emphasizing the global behavior of the metabolomic fingerprint, here we focus on several series of metabolites spanning over extensive polarity, concentration, and molecular mass ranges., (Copyright 2010 Elsevier Inc. All rights reserved.)

Alkylating agents occur in the environment and are formed endogenously. Tobacco smoke contains a variety of alkylating agents or precursors including, among others, N-nitrosodimethylamine (NDMA), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), acrylonitrile and ethylene oxide. We developed and validated a method for the simultaneous determination of methylmercapturic acid (MMA, biomarker for methylating agents such as NDMA and NNK), 2-hydroxyethylmercapturic acid (HEMA, biomarker for ethylene oxide) and 2-cyanoethylmercapturic acid (CEMA, biomarker for acrylonitrile) in human urine using deuterated internal standards of each compound. The method involves liquid/liquid extraction of the urine sample, solid phase extraction on anion exchange cartridges, derivatization with pentafluorobenzyl bromide (PFBBr), liquid/liquid extraction of the reaction mixture and LC-MS/MS analysis with positive electrospray ionization. The method was linear in the ranges of 5.00-600, 1.00-50.0 and 1.50-900 ng/ml for MMA, HEMA and CEMA, respectively. The method was applied to two clinical studies in adult smokers of conventional cigarettes who either continued smoking conventional cigarettes, were switched to test cigarettes consisting of either an electrically heated cigarette smoking system (EHCSS) or having a highly activated carbon granule filter that were shown to have reduced exposure to specific smoke constituents, or stopped smoking. Urinary excretion of MMA was found to be unaffected by switching to the test cigarettes or stop smoking. Urinary HEMA excretion decreased by 46 to 54% after switching to test cigarettes and by approximately 74% when stopping smoking. Urinary CEMA excretion decreased by 74-77% when switching to test cigarettes and by approximately 90% when stopping smoking. This validated method for urinary alkylmercapturic acids is suitable to distinguish differences in exposure not only between smokers and nonsmokers but also between smoking of conventional and the two test cigarettes investigated in this study., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Tobacco-specific N-nitrosamines (TSNA) include 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), N-nitrosonornicotine (NNN), N-nitrosoanabasine (NAB) and N-nitrosoanatabine (NAT). TSNA are suggested to play an important role in tobacco smoke carcinogenesis. We have developed and validated an LC-MS/MS method for the determination of total (free and conjugated) TSNA in human urine. The limits of detection (LOD) were 2.0, 0.8, 1.1 and 0.7 pg/ml for NNAL, NNN, NAB and NAT, respectively. Smokers were found to have significantly higher levels of TSNA in their urine than nonsmokers. In conclusion, the newly developed method is suitable for assessing the tobacco use-related exposure to NNK, NNN, NAB and NAT.

This article describes the generation of the Human Combinatorial Antibody Library HuCAL GOLD. HuCAL GOLD is a synthetic human Fab library based on the HuCAL concept with all six complementarity-determining regions (CDRs) diversified according to the sequence and length variability of naturally rearranged human antibodies. The human antibody repertoire was analyzed in-depth, and individual CDR libraries were designed and generated for each CDR and each antibody family. Trinucleotide mixtures were used to synthesize the CDR libraries in order to ensure a high quality within HuCAL GOLD, and a beta-lactamase selection system was employed to eliminate frame-shifted clones after successive cloning of the CDR libraries. With these methods, a large, high-quality library with more than 10 billion functional Fab fragments was achieved. By using CysDisplay, the antibody fragments are displayed on the tip of the phage via a disulfide bridge between the phage coat protein pIII and the heavy chain of the antibody fragment. Efficient elution of specific phages is possible by adding reducing agents. HuCAL GOLD was challenged with a variety of different antigens and proved to be a reliable source of high-affinity human antibodies with best affinities in the picomolar range, thus functioning as an excellent source of antibodies for research, diagnostic, and therapeutic applications. Furthermore, the data presented in this article demonstrate that CysDisplay is a robust and broadly applicable display technology even for high-throughput applications.

Androgens are considered to play a substantial role in pathogenesis of both benign prostatic hyperplasia (BPH) and prostate cancer. The importance of determination of androgen levels in tissue and serum for cancer progression and prognosis has been poorly understood. The aim of study was to find out hormonal differences in both diseases, their correlations between intraprostatic and serum levels and predicted value of their investigation. Testosterone, dihydrotestosterone, androstenedione and also epitestosterone were determined in prostate tissue from 57 patients who underwent transvesical prostatectomy for BPH and 121 patients after radical prostatectomy for prostate cancer. In 75 subjects with cancer and 51 with BPH the serum samples were analyzed for testosterone, dihydrotestosterone and SHBG. Significantly higher intraprostatic androgen concentrations, i.e. 8.85+/-6.77 versus 6.44+/-6.43 pmol/g, p<0.01 for dihydrotestosterone, and 4.61+/-7.02 versus 3.44+/-4.53 pmol/g, p<0.05 for testosterone, respectively, were found in patients with prostate cancer than in BPH. Higher levels in cancer tissue were found also for epitestosterone. However, no differences were found in serum levels. Highly significant correlations occurred between all pairs of intraprostatic androgens and also epitestosterone as well as between serum testosterone and dihydrotestosterone (p<0.001) in both BPH and cancer groups. Correlation was not found between corresponding tissue and serum testosterone and dihydrotestosterone, either in benign or cancer samples. The results point to importance of intraprostatic hormone levels for evaluation of androgen status of patients, contrasting to a low value of serum hormone measurement.

In order to determine whether smokers of cigarettes in the contemporary yield ranges of the German market (0.1-1.0mg nicotine, 1-10mg tar) differ in their actual exposure to various smoke constituents, we performed a field study with 274 smokers and 100 non-smokers. The following biomarkers were determined: In 24-h urine: Nicotine equivalents (molar sum of nicotine, cotinine, trans-3'-hydroxycotinine and their respective glucuronides), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL, metabolite of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, NNK), 3-hydroxypropylmercapturic acid (metabolite of acrolein), trans,trans-muconic acid, S-phenylmercapturic acid (metabolites of benzene), 1-hydroxypyrene (metabolite of pyrene); in saliva: Cotinine and trans-3'-hydroxycotinine; in exhaled air: Carbon monoxide; in blood: Methyl-, hydroxyethyl-, cyanoethyl- (biomarker of acrylonitrile) and carbamoylethylvaline (biomarker of acrylamide) hemoglobin adducts. All biomarkers were found to be significantly higher in smokers compared to non-smokers and showed strong correlations with the daily cigarette consumption. Biomarker levels and per cigarette increases in smokers were at most weakly related to the machine-derived smoke yields. It is concluded that machine-derived yields of cigarettes from the contemporary German cigarette market have little or no impact on the actual smoking-related exposure determined by suitable biomarkers.

Runx2 is an essential factor for skeletogenesis and heterozygous loss causes cleidocranial dysplasia in humans and a corresponding phenotype in the mouse. Homozygous Runx2-deficient mice lack hypertrophic cartilage and bone. We compared the expression profiles of E14.5 wildtype and Runx2(-/-) murine embryonal humeri to identify new transcripts potentially involved in cartilage and bone development. Seventy-one differentially expressed genes were identified by two independent oligonucleotide-microarray hybridizations and quantitative RT-PCR experiments. Gene Ontology analysis demonstrated an enrichment of the differentially regulated genes in annotations to terms such as extracellular, skeletal development, and ossification. In situ hybridization on E15.5 limb sections was performed for all 71 differentially regulated genes. For 54 genes conclusive in situ hybridization results were obtained and all of them showed skeletal expression. Co-expression with Runx2 was demonstrated for 44 genes. While 41 of the 71 differentially expressed genes have a known role in bone and cartilage, we identified 21 known genes that have not yet been implicated in skeletal development and 9 entirely new transcripts. Expression in the developing skeleton was demonstrated for 21 of these genes.

The major urinary metabolites of 1,3-butadiene are monohydroxybutenyl-mercapturic acids (MHBMA) and dihydroxy-butyl-mercapturic acid (DHBMA). These metabolites can be used as biomarkers of exposure to this diene. In order to determine the smoking-related exposure to 1,3-butadiene, we have developed a rapid LC-MS/MS method for the determination of MHBMA and DHBMA in urine of humans and rats. The method requires 2-5 ml of urine which is solid phase extracted prior to LC-MS/MS analysis. Precision for MHBMA is < or =11.2% for human and < or = 17% for rat urine. Corresponding values for DHBMA are < or = 7.2 and < or = 19%, respectively. Recovery rates are approximately 100% for both analytes in human urine and about 115% in rat urine. Limits of detection (LOD) are for humans 0.9 and 23 ng/ml and for rats 1.5 and 33 ng/ml for MHBMA and DHBMA, respectively. Application of the method to urine of humans and rats showed a significant effect of tobacco smoke exposure on the urinary excretion of MHBMA and the metabolic ratio DHBMA/(DHBMA + MHBMA).

Unlabelled: In the last few years it has been proved that risk factors for atherosclerosis are present in children and adolescents, and that already at this early age they are connected with anatomic, atheromatous changes in vessels. These changes can not be fully explained as occurring in young people exhibiting traditional risk factors for the disease. The aim of the study was to evaluate levels of several new atherosclerosis risk factors (lipoprotein (a) (Lp(a)), apolipoprotein A-I (Apo A-I), apolipoprotein B (Apo B), homocysteine (Hcy), fibrinogen (FB), tissue plasminogen activator (t-PA) and tissue plasminogen activator inhibitor type 1 (PAI-1)) in children and adolescents with traditional risk factors (obesity, hypertension, diabetes)., Materials and Methods: The study group consisted of 285 children and adolescents aged 14.3 years. Children were divided according to their main disease into groups: group A, children with obesity (n=49); group B, children with obesity and coexisting hypertension (n=56); group C, children with hypertension (n=58) and group D, children with diabetes (n=122). Control group consisted of 79 healthy children and adolescents aged 14.1 years. Lp(a), Apo A-I and Apo B levels were estimated by use of immunoturbidimetric methods; total Hcy, FB, t-PA and PAI-1 were estimated by use of immunoenzymatic methods., Results: Lp(a) level in the total study group was 30 mg/dl and was over twice higher than in control group, 14 mg/dl. Apo A-I level was significantly lower in group A (127.6 mg/dl) and in group B (125.8 mg/dl) versus 135.6 mg/dl in controls. The level of Apo B was significantly higher in total study group (86.2 mg/dl) and in groups A, B and D versus 73.5 mg/dl in controls. Hcy was higher in group B (8 micromol/l) and in group C (9.4 micromol/l) versus 6.2 micromol/l in the control group. The FB level was higher in the total study group (276.7 mg/dl) and in groups A (318.8 mg/dl) and B (322.6 mg/dl) versus 252.8 mg/dl in controls. Significantly higher t-PA level was found in groups A (9 ng/ml) and B (9.7 ng/ml) versus 7.3 ng/ml in controls, and PAI-1 level was significantly higher in the total study group (62.3 ng/ml) and in groups A (73.8 ng/ml), B (78 ng/ml) and C (73 ng/ml) versus 42.4 ng/ml in the control group. Correlation analysis showed significant relationship between body mass index (BMI) and Apo B, Hcy, FB, t-PA and PAI-1. Blood pressure values correlated positively with Hcy. Correlations were verified in multiple regression analysis models: FB and t-PA levels depended on BMI, and Hcy depended on systolic blood pressure., Conclusions: (1) Young obese, hypertensive and diabetic patients present significant disturbances in lipid metabolism, regarding mainly total cholesterol, LDL, triglycerides, as well as Lp(a), Apo A-I and Apo B levels. Unfavourable lipid profile is characteristic mainly for children with obesity and accompanying hypertension. (2) Elevated Hcy levels are found in children with hypertension. (3) Elevated FB level and diminished fibrinolytic activity are characteristic of obese children.

Heterotopic adrenal tissue is not uncommon, especially in the urogenital system. Adrenocortical tissue in the placenta, however, is presumably very rare. To our knowledge, four cases have been published. There are several different theories to explain such a heterotopia. According to our findings, an embolic spread of adrenal precursor cells via fetal vascular shortcuts is the most likely mechanism. Apart from that hypothesis, the possibility of a monodermal teratoma as well as of an aberrant differentiation of cells of the extraembryonic mesoderm are considered in the literature., (Copyright 2000 Harcourt Publishers Ltd.)

Small gap junction plaques are associated with tight junction strands in some cell types including hepatocytes and it is thought that they may be closely related to tight junctions and the establishment of cell polarity. In order to examine roles of gap junctions in regulating expression and structure of tight junctions, we transfected human Cx32 cDNA into immortalized mouse hepatocytes (CHST8 cells) which lack endogenous Cx32 and Cx26. Immunocytochemistry revealed that endogenous integral tight junction protein occludin was strongly localized and was colocalized with Cx32 at cell borders in transfectants, whereas neither was detected in parental cells. In Northern blots, mRNAs encoding occludin and the other integral tight junction proteins, claudin-1 and -2, were induced in the transfectants compared to parental cells. In Western blots, occludin protein was increased in the transfectants compared to parental cells, and binding of occludin to Cx32 protein was demonstrated by immunoprecipitation. In freeze fracture of the transfectants, tight junction strands were more numerous and complex compared to parental cells, and small gap junction plaques appeared within induced tight junction strands. Nevertheless, no change in barrier function of tight junctions was observed. These results indicate that in hepatocytes, gap junction, and tight junction expression are closely coordinated, and that Cx32 may play a role in regulating occludin expression., (Copyright 1999 Academic Press.)

This case report describes the development of a leiomyosarcoma in the chest wall of a patient previously treated for Hodgkin's disease. Although a similar case has not been reported previously, we managed our patient with wide local excision.

In the last two decades many countries started national or regional surveys to assess the average exposure to natural radiation of the general public. Survey programmes for external radiation as well as for radon and decay products have been performed are still in progress or will be started in the near future. The paper discusses briefly technical and organisational aspects of survey measurements as well as practical experiences and results available from national or regional survey programmes.