Outcome groups and a practical tool to predict success of shock wave lithotripsy in daily clinical routine.

PURPOSE: To improve outcome prediction of extracorporeal shock wave lithotripsy (SWL) by development of a model based on easily available clinical and radiographical predictors and suitable for daily clinical use. MATERIALS AND METHODS: We evaluated predictive factors for SWL success in 517 consecutive patients suffering from urinary calculi who underwent SWL between 2010 and 2018. Analyses included descriptive statistics, receiver operating characteristic statistics and logistic regression. Predictive value was improved by combining parameters using model selection and recursive partitioning. RESULTS: Of the 517 patients, 310 (60.0%) had a successful SWL. Best individual predictor of SWL success was mean attenuation (MAV), with an area under the curve (AUC) of 0.668, and an optimal cutpoint (OC) of 987.5 HU. The best multivariable model, including MAV, stone size, skin to stone distance (SSD), presence of an indwelling stent, and four interaction effects, yielded an AUC of 0.736. Recursive partitioning would categorize patients into three outcome groups with high (76.9%), intermediate (41%) and low (10%) success probability. High probability of SWL success (76.9%) was found for patients with a stone with MAV ≤ 987 HU or with MAV > 987 HU but stone size ≤ 11 mm and SSD (45°) ≤ 88 mm. CONCLUSION: A model based on four established predictors, and provided as an Excel®-Tool, can clearly improve prediction of SWL success. In addition, patients can be classified into three defined outcome groups based on simple cutpoint combinations. Both tools improve informed decision-making in daily clinical practice and might reduce failure rates.

SH2B3 aberrations enriched in iAMP21 B lymphoblastic leukemia.

Acute lymphoblastic leukemia (ALL) represents the most common childhood malignancy. Although survival for pediatric B-ALL has approached 90%, variability in outcome among and within cytogenetically defined subgroups persists. While G-banding and fluorescence in situ hybridization (FISH) have been used to characterize leukemic clones, there is added value of chromosomal microarray and next generation sequencing in screening genome-wide for copy number aberrations, copy neutral loss of heterozygosity and nucleotide variations. Evaluation of novel genetic aberrations can provide information about the biologic mechanisms of cytogenetically defined subgroups associated with poor prognosis, explain heterogeneity in patient outcome and identify novel targets for therapeutic intervention. The high risk B-ALL intrachromosomal amplification of chromosome 21, (iAMP21), subtype is characterized by amplification of a region of chromosome 21 that typically encompasses the RUNX1 gene and is associated with poor prognosis. Analysis of chromosomal microarray and FISH data revealed that deletions of SH2B3, encoding a negative regulator of multiple tyrosine kinase and cytokine signaling pathways, are enriched among leukemias harboring iAMP21. Enrichment of SH2B3 aberrations in the iAMP21 subtype may indicate that loss of SH2B3 contributes to disease progression and raises the possibility that these leukemias may be sensitive to tyrosine kinase inhibitors.

Multicenter validation of cancer gene panel-based next-generation sequencing for translational research and molecular diagnostics.

The simultaneous detection of multiple somatic mutations in the context of molecular diagnostics of cancer is frequently performed by means of amplicon-based targeted next-generation sequencing (NGS). However, only few studies are available comparing multicenter testing of different NGS platforms and gene panels. Therefore, seven partner sites of the German Cancer Consortium (DKTK) performed a multicenter interlaboratory trial for targeted NGS using the same formalin-fixed, paraffin-embedded (FFPE) specimen of molecularly pre-characterized tumors (n = 15; each n = 5 cases of Breast, Lung, and Colon carcinoma) and a colorectal cancer cell line DNA dilution series. Detailed information regarding pre-characterized mutations was not disclosed to the partners. Commercially available and custom-designed cancer gene panels were used for library preparation and subsequent sequencing on several devices of two NGS different platforms. For every case, centrally extracted DNA and FFPE tissue sections for local processing were delivered to each partner site to be sequenced with the commercial gene panel and local bioinformatics. For cancer-specific panel-based sequencing, only centrally extracted DNA was analyzed at seven sequencing sites. Subsequently, local data were compiled and bioinformatics was performed centrally. We were able to demonstrate that all pre-characterized mutations were re-identified correctly, irrespective of NGS platform or gene panel used. However, locally processed FFPE tissue sections disclosed that the DNA extraction method can affect the detection of mutations with a trend in favor of magnetic bead-based DNA extraction methods. In conclusion, targeted NGS is a very robust method for simultaneous detection of various mutations in FFPE tissue specimens if certain pre-analytical conditions are carefully considered.

Exploring the science of thinking independently together: Faraday Discussion Volume 204 - Complex Molecular Surfaces and Interfaces, Sheffield, UK, July 2017.

The 2017 Faraday Discussion on Complex Molecular Surfaces and Interfaces brought together theoreticians and experimentalists from both physical and chemical backgrounds to discuss the relevant applied and fundamental research topics within the broader field of chemical surface analysis and characterization. Main discussion topics from the meeting included the importance of "disordered" two-dimensional (2D) molecular structures and the utility of kinetically trapped states. An emerging need for new experimental tools to address dynamics and kinetic pathways involved in self-assembled systems, as well as the future prospects and current limitations of in silico studies were also discussed. The following article provides a brief overview of the work presented and the challenges discussed during the meeting.

Induction of the inflammatory regulator A20 by gibberellic acid in airway epithelial cells.

BACKGROUND AND PURPOSE: NF-κB-driven inflammation is negatively regulated by the zinc finger protein A20. Gibberellic acid (GA3 ) is a plant-derived diterpenoid with documented anti-inflammatory activity, which is reported to induce A20-like zinc finger proteins in plants. Here, we sought to investigate the anti-inflammatory effect of GA3 in airway epithelial cells and determine if the anti-inflammatory action relates to A20 induction. EXPERIMENTAL APPROACH: Primary nasal epithelial cells and a human bronchial epithelial cell line (16HBE14o-) were used. Cells were pre-incubated with GA3 , stimulated with Pseudomonas aeruginosa LPS; IL-6 and IL-8 release, A20, NF-κB and IκBα expression were then evaluated. To determine if any observed anti-inflammatory effect occurred via an A20-dependent mechanism, A20 was silenced using siRNA. KEY RESULTS: Cells pre-incubated with GA3 had significantly increased levels of A20 mRNA (4 h) and protein (24 h), resulting in a significant reduction in IL-6 and IL-8 release. This effect was mediated via reduced IκBα degradation and reduced NF-κB (p65) expression. Furthermore, the anti-inflammatory action of GA3 was abolished in A20-silenced cells. CONCLUSIONS AND IMPLICATIONS: We showed that A20 induction by GA3 attenuates inflammation in airway epithelial cells, at least in part through its effect on NF-κB and IκBα. GA3 or gibberellin-derived derivatives could potentially be developed into anti-inflammatory drugs for the treatment of chronic inflammatory diseases associated with A20 dysfunction.

Consultas ambulatorias pediátricas atendidas en el servicio de urgencia de un hospital universitario / Pediatric outpatient consultation at the emergency department of a university hospital

Introducción: Conocer el perfil epidemiológico de las consultas pediátricas atendidas en el servicio de urgencia (SU) es esencial para planificar los procesos de atención médica y orientar los programas de educación e investigación. Objetivos: Describir las características de la población infantil y los principales motivos de consulta (MC) atendidos en un SU pediátrico. Pacientes y Método: Estudio clínico descriptivo retrospectivo de las visitas realizadas a un SU infantil en un hospital académico de Santiago durante un período de doce meses. Se analizaron los MC por grupo etario, gravedad, estacionalidad, forma de egreso y frecuencia de visitas recurrentes. Resultados: Se evaluaron 24.531 consultas pediátricas, 51,9 por ciento (n = 12.720) eran varones. La edad de los pacientes osciló entre un día y 15 años, con una mediana de 36,5 meses. El 1,5 por ciento de los pacientes (n = 362) eran recién nacidos (RN), 17,6 por ciento (n = 4.326) lactantes, 51,9 por ciento (n = 12.725) preescolares y 29 por ciento (n = 7.118) escolares. Los principales MC fueron fiebre (n = 6.643, 28,2 por ciento), síntomas gastrointestinales (n = 5.606, 23,8 por ciento) y síntomas respiratorios (n = 5.018, 21,3 por ciento), los cuales no difirieron significativamente según género. La mayoría de los pacientes (95,5 por ciento) fueron enviados a su domicilio. El riesgo de hospitalización fue más elevado en los RN y en aquellos que consultaron por ictericia (OR = 7,20; IC 95 por ciento 3,12-16,6), síntomas neurológicos (OR = 6,90; IC 95 por ciento 4,60-10,4) e intoxicaciones (OR = 6,45; IC 95 por ciento 2,82-14,7). Alrededor del 4 por ciento fueron consultas repetidas, especialmente en los RN. Conclusiones: El perfil epidemiológico de las consultas pediátricas atendidas en el SU fue similar al descrito en estudios internacionales. Sin embargo, encontramos una menor tasa de hospitalización a pesar que los pacientes presentaban un perfil de riesgo similar en la admisión al SU.

Introduction: To determine the epidemiological profile of pediatric consultations treated at the emergency department (ED) is essential for planning processes of medical care and to guide education programs and research. Objectives: To describe the characteristics of the child population and the main reasons for consultation (RFC) seen in a pediatric emergency service. Patients and Method: A retrospective, descriptive clinical study was conducted regarding the visits to the Children's Emergency Service of an academic hospital in Santiago, for a period of twelve months. RFC were analyzed by age group, severity, seasonality, disposition and frequency of recurrent visits. Results: 24,531 pediatric consultations were evaluated, 51.9 percent were male (n = 12,720). The age of the patients ranged between one day old and 15 years, with a median age of 36.5 months. 1.5 percent of patients were newborns (NB), 17.6 percent were infants (n = 4,326), 51.9 percent were preschoolers (n = 12,725) and 29 percent were school children (n = 7,118). Major RFC were fever (n = 6,643, 28.2 percent), gastrointestinal symptoms (n = 5,606, 23.8 percent) and respiratory symptoms (n = 5,018, 21.3 percent), which did not differ significantly according to gender. Most patients (95.5 percent) were sent to their homes. The risk of hospitalization was more elevated in NB and in those with jaundice (OR = 7.20, 95 percent CI 3.12 to 16.6), neurological symptoms (OR = 6.90, 95 percent CI 4.60 -10.4) and poisoning (OR = 6.45, 95 percent CI 2.82 to 14.7). About 4 percent were repeat visits, especially in the NB group. Conclusions: The epidemiological profile of pediatric consultations seen at the ED was similar to that described in previous studies. However, a lower rate of hospitalization was found even though the patients had similar risk profile.

[Pediatric outpatient consultation at the emergency department of a university hospital]. / Consultas ambulatorias pediátricas atendidas en el Servicio de Urgencia de un hospital universitario.

INTRODUCTION: To determine the epidemiological profile of pediatric consultations treated at the emergency department (ED) is essential for planning processes of medical care and to guide education programs and research. OBJECTIVES: To describe the characteristics of the child population and the main reasons for consultation (RFC) seen in a pediatric emergency service. PATIENTS AND METHOD: A retrospective, descriptive clinical study was conducted regarding the visits to the Children's Emergency Service of an academic hospital in Santiago, for a period of twelve months. RFC were analyzed by age group, severity, seasonality, disposition and frequency of recurrent visits. RESULTS: 24,531 pediatric consultations were evaluated, 51.9% were male (n=12,720). The age of the patients ranged between one day old and 15 years, with a median age of 36.5 months. 1.5% of patients were newborns (NB), 17.6% were infants (n=4,326), 51.9% were preschoolers (n=12,725) and 29% were school children (n=7,118). Major RFC were fever (n=6,643, 28.2%), gastrointestinal symptoms (n=5,606, 23.8%) and respiratory symptoms (n=5,018, 21.3%), which did not differ significantly according to gender. Most patients (95.5%) were sent to their homes. The risk of hospitalization was more elevated in NB and in those with jaundice (OR=7.20, 95% CI 3.12 to 16.6), neurological symptoms (OR=6.90, 95% CI 4.60 -10.4) and poisoning (OR=6.45, 95% CI 2.82 to 14.7). About 4% were repeat visits, especially in the NB group. CONCLUSIONS: The epidemiological profile of pediatric consultations seen at the ED was similar to that described in previous studies. However, a lower rate of hospitalization was found even though the patients had similar risk profile.

Exogenous hormone use, reproductive history and risk of adult myeloid leukaemia.

BACKGROUND: A hormonal aetiology is one explanation for the lower incidence of myeloid leukaemia in women compared with men. METHODS: In this population-based case-control study, we evaluated associations between exogenous hormone use and reproductive history and myeloid leukaemia, overall and by disease subtype. RESULTS: We observed a suggestive association between oral contraceptive use and acute myeloid leukaemia (odds ratio=0.55, 95% confidence interval=0.32-0.96). Hormone replacement therapy and reproductive factors were not associated with risk. CONCLUSION: Despite the biological plausibility for a role of oestrogen in leukaemogenesis, other aetiologic factors are likely to explain the differing incidence rates in males and females.

Three-dimensional kinematic stress magnetic resonance image analysis shows promise for detecting altered anatomical relationships of tissues in the cervical spine associated with painful radiculopathy.

For some patients with radiculopathy a source of nerve root compression cannot be identified despite positive electromyography (EMG) evidence. This discrepancy hampers the effective clinical management for these individuals. Although it has been well-established that tissues in the cervical spine move in a three-dimensional (3D) manner, the 3D motions of the neural elements and their relationship to the bones surrounding them are largely unknown even for asymptomatic normal subjects. We hypothesize that abnormal mechanical loading of cervical nerve roots during pain-provoking head positioning may be responsible for radicular pain in those cases in which there is no evidence of nerve root compression on conventional cervical magnetic resonance imaging (MRI) with the neck in the neutral position. This biomechanical imaging proof-of-concept study focused on quantitatively defining the architectural relationships between the neural and bony structures in the cervical spine using measurements derived from 3D MR images acquired in neutral and pain-provoking neck positions for subjects: (1) with radicular symptoms and evidence of root compression by conventional MRI and positive EMG, (2) with radicular symptoms and no evidence of root compression by MRI but positive EMG, and (3) asymptomatic age-matched controls. Function and pain scores were measured, along with neck range of motion, for all subjects. MR imaging was performed in both a neutral position and a pain-provoking position. Anatomical architectural data derived from analysis of the 3D MR images were compared between symptomatic and asymptomatic groups, and the symptomatic groups with and without imaging evidence of root compression. Several differences in the architectural relationships between the bone and neural tissues were identified between the asymptomatic and symptomatic groups. In addition, changes in architectural relationships were also detected between the symptomatic groups with and without imaging evidence of nerve root compression. As demonstrated in the data and a case study the 3D stress MR imaging approach provides utility to identify biomechanical relationships between hard and soft tissues that are otherwise undetected by standard clinical imaging methods. This technique offers a promising approach to detect the source of radiculopathy to inform clinical management for this pathology.

AR intragenic deletions linked to androgen receptor splice variant expression and activity in models of prostate cancer progression.

Reactivation of the androgen receptor (AR) during androgen depletion therapy (ADT) underlies castration-resistant prostate cancer (CRPCa). Alternative splicing of the AR gene and synthesis of constitutively active COOH-terminally truncated AR variants lacking the AR ligand-binding domain has emerged as an important mechanism of ADT resistance in CRPCa. In a previous study, we demonstrated that altered AR splicing in CRPCa 22Rv1 cells was linked to a 35-kb intragenic tandem duplication of AR exon 3 and flanking sequences. In this study, we demonstrate that complex patterns of AR gene copy number imbalances occur in PCa cell lines, xenografts and clinical specimens. To investigate whether these copy number imbalances reflect AR gene rearrangements that could be linked to splicing disruptions, we carried out a detailed analysis of AR gene structure in the LuCaP 86.2 and CWR-R1 models of CRPCa. By deletion-spanning PCR, we discovered a 8579-bp deletion of AR exons 5, 6 and 7 in the LuCaP 86.2 xenograft, which provides a rational explanation for synthesis of the truncated AR v567es AR variant in this model. Similarly, targeted resequencing of the AR gene in CWR-R1 cells led to the discovery of a 48-kb deletion in AR intron 1. This intragenic deletion marked a specific CWR-R1 cell population with enhanced expression of the truncated AR-V7/AR3 variant, a high level of androgen-independent AR transcriptional activity and rapid androgen independent growth. Together, these data demonstrate that structural alterations in the AR gene are linked to stable gain-of-function splicing alterations in CRPCa.

Importancia actual de la vitamina D: déficit prevalente y asociación a diversas patologías / Current importance of vitamin D

En los últimos años se ha detectado una epidemia mundial de bajos niveles de vitamina D en la población general, la que ha alertado sobre su correlación con diversas enfermedades. Hay muchas razones que pueden ser las causales de este importante problema, siendo la baja exposición a luz solar uno de los principales. Nuestra especialidad juega un rol central en promover una foto exposición más saludable o suplementando a pacientes de alto riesgo para lograr niveles adecuados de esta vitamina. En nuestra revisión analizamos brevemente las causas de bajos niveles de vitamina D, sus funciones y las recientes correlaciones de su déficit con varias condiciones, entre ellas; el cáncer de piel, atopia y enfermedades autoinmunes. Finalmente recomendamos búsqueda activa y suplementación cuando sea necesario.

In the last few years, a worldwide epidemic of low levels of vitamin D in general population has been discovered, raising an alert about its correlation with health problems. Many reasons may be responsible for this important problem, and insufficient exposure to sunlight may be the main one. Our specialty has a key role in promoting a healthy sun exposure or supplementing high risk patients, to keep adequate levels of this vitamin. In our review, we analyze briefly the causes for low levels of vitamin D, its functions and newly discovered correlations with diseases, including skin cancer, atopy and autoimmune conditions. Finally we recommend screening and supplementation, when necessary.

Bocavirus humano en Chile: características clínicas y epidemiológicas en niños con infecciones respiratorias / Human bocavirus in Chile: clinical characteristics and epidemiological profile in children with acute respiratory tract infections

Background: Human bocavirus (HBoV) is a newly discovered parvovirus found in children with acute respiratory tract infections (ARTI). Objectives: To describe the epidemiological and clinical profile of children < 5 years old consulting for ARTI, comparing cases of HBoV monoinfection and coinfection with other known respiratory viruses. Furthermore, we aimed to estimate the prevalence of viral shedding in asymptomatic children and perform phylogenetic analysis. Patients and Methods: We investigated the presence of HBoV in nasopharyngeal secretions from children consulting for AlRTI and among asymptomatic controls, between 2007 and 2008, by polymerase chain reaction. Results: HBoV was detected in 79 (21.8 percent) of 362 nasopharyngeal swabs obtained from children with ARTI. In 60/79 (76 percent), coinfection with other respiratory viruses was confirmed. Most common symptoms were cough, fever and rhinorrhea. Children infected only with HBoV showed significantly lower frequencies of respiratory distress, oxygen requirements and hospital admission than those with coinfection. HBoV was detected in 6/16 (37.5 percent) samples from asymptomatic children. The phylogenetic analysis of viruses from Chilean patients revealed that circulating HBoV was closely related to original strains. Conclusions: HBoV was found either in symptomatic and asymptomatic children. The severity of the disease was greater when HBoV was associated to other respiratory viruses.

Introducción: Bocavirus humano (HBoV) es un nuevo parvovirus encontrado en niños con infecciones respiratorias agudas (IRA). Objetivos: Describir la epidemiología y perfil clínico en niños < 5 años con IRA, comparando aquellos con HBoV como único agente identificado, con los que tenían co-infección con otro virus respiratorio. Además se evaluó su prevalencia en niños asintomáticos, y se realizó análisis filogenético. Materiales y Métodos: Se investigó la presencia de HBoV, por medio de reacción de polimerasa en cadena, en muestras de secreción nasofaríngea obtenida en niños con IRA y en controles asintomáticos, entre 2007 y 2008. Resultados: Se detectó HBoV en 79 (21,8 por ciento) de 362 muestras obtenidas en pacientes con IRA. En 60/79 (76 por ciento), se demostró co-infección. Los síntomas más frecuentes fueron tos, fiebre y rinorrea. Los pacientes con HBoV como único agente identificado mostraron frecuencias significativamente menores de dificultad respiratoria, requerimiento de oxígeno y hospitalización, comparado con los co-infectados. HBoV se detectó en 6/16 (37,5 por ciento) muestras de niños asintomáticos. El análisis filogenético de las cepas chilenas demuestra estrecha relación con las cepas originales. Conclusiones: HBoV está presente en niños chilenos con IRA y asintomáticos. La gravedad de la enfermedad fue mayor en el grupo con co-infección.

[Kawasaki disease: a clinical serie]. / Enfermedad de Kawasaki: una serie clínica.

INTRODUCTION: Kawasaki disease (KD) is a serious disease in children due to its potential complications and sequelae if not promptly and adequately managed. OBJECTIVES: To describe clinical and epidemiological characteristics of children hospitalized due to KD at a tertiary care center and identify risk factors for poor outcome. PATIENTS AND METHODS: Retrospective and descriptive study of 32 medical records of patients hospitalized with diagnosis of KD at a tertiary care center of Santiago, Chile between February 1999 and May 2007. RESULTS: The annual frequency was of 5 cases, mainly boys and during spring. The median age at diagnosis was 1.5 years and 87.5% of the children were younger than 5 years. Typical presentation prevailed in all ages (68,7%). Coronary artery affection, including dilatation or aneurisms, occurred in 21.9% of the cases, with aneurysms in 3 cases. All patients were treated with aspirin and intravenous immunoglobulin (IVIG); 4 patients required a second dose. No deaths were reported. The identified risk factors for poor outcome were age older than 5 years and hypoalbuminemia. CONCLUSIONS: KD is an infrequent disease that mainly occurs in children younger than 5 years and with a typical presentation. There are risk factors associated with poor outcome.

Enfermedad de Kawasaki: una serie clínica / Kawasaki disease: a clinical serie

Introduction: Kawasaki disease (KD) is a serious disease in children due to its potential complications and sequelae if not promptly and adequately managed. Objectives: To describe clinical and epidemiological characteristics of children hospitalized due to KD at a tertiary care center and identify risk factors for poor outcome. Patients and Methods: Retrospective and descriptive study of 32 medical records of patients hospitalized with diagnosis of KD at a tertiary care center of Santiago, Chile between February 1999 and May 2007. Results: The annual frequency was of 5 cases, mainly boys and during spring. The median age at diagnosis was 1.5 years and 87.5 percent of the children were younger than 5 years. Typical presentation prevailed in all ages (68,7 percent). Coronary artery affection, including dilatation or aneurisms, occurred in 21.9 percent of the cases, with aneurysms in 3 cases. All patients were treated with aspirin and intravenous immunoglobulin (IVIG); 4 patients required a second dose. No deaths were reported. The identified risk factors for poor outcome were age older than 5 years and hypoalbumin-emia. Conclusions: KD is an infrequent disease that mainly occurs in children younger than 5 years and with a typical presentation. There are risk factors associated with poor outcome.

Introducción: La enfermedad de Kawasaki (EK) es una afección grave en pediatría, por sus eventuales complicaciones y secuelas si no es tratada adecuada y precozmente. Objetivos: Describir el perfil clínico-epidemiológico de los niños hospitalizados en una clínica de la Región Metropolitana e identificar factores de riesgo para evolución desfavorable. Pacientes y Métodos: Estudio descriptivo, retrospectivo, de 32 pacientes hospitalizados en la Clínica Santa María con diagnóstico de egreso de EK entre febrero de 1999 y mayo de 2007. Resultados: La frecuencia anual fue de 5 casos, con predominio en primavera y en el sexo masculino. La mediana para la edad fue de 1,5 años, 87,5 por ciento tenían menos de 5 años. Predominó la presentación típica (68,7 por ciento). Hubo dilatación y/o aneurisma de arterias coronarias en 21,9 por ciento, con 3 casos de aneurismas. El tratamiento se realizó con inmunoglobulina intravenosa (IGIV) y ácido acetilsalicílico. Cuatro pacientes requirieron una segunda dosis de IGIV. No hubo fallecidos. Como factores de riesgo para mala evolución se encontraron la edad de presentación sobre 5 años y la hipoalbuminemia. Conclusiones: La EK es una patología poco frecuente, predominante en niños bajo 5 años de edad y con presentación típica. Sugieren evolución de mayor gravedad la presentación sobre los 5 años y la hipoalbuminemia a cualquier edad.

LAPTM4A interacts with hOCT2 and regulates its endocytotic recruitment.

Human organic cation transporter 2 (hOCT2) is involved in the transport of endogenous and exogenous organic cations mainly in cells of the kidney and the brain. Here, we focus on the regulation of hOCT2 by direct protein-protein interaction. Screening within a mating-based split-ubiquitin-yeast-two-hybrid system (mBSUS) revealed the lysosomal-associated protein transmembrane 4 alpha (LAPTM4A) as a potential interacting protein. Interaction of LAPTM4A and hOCT2 was confirmed by pulldown assays, FRET microscopy analysis and immunofluorescence microscopy. Functionally, overexpression of LAPTM4A significantly decreased ASP(+) uptake in HEK293 cells stably transfected with hOCT2, suggesting a negative regulation of hOCT2-mediated transport. Furthermore, overexpression of LAPTM4A leads to a significantly decreased hOCT2 plasma membrane expression in surface biotinylation experiments. In addition, significant expression of LAPTM4A in human kidney was demonstrated by immunoblotting and immunofluorescence.In this work, LAPTM4A has been identified as interaction partner of hOCT2. LAPTM4A regulates the function of hOCT2 by influencing its trafficking to/from the cell membrane and processing it via the intracellular sorting machinery.

Assessment strategies and decision criteria for pesticides with endocrine disrupting properties relevant to humans.

There is growing concern that environmental substances with a potential to modulate the hormonal system may have harmful effects on human health. Consequently, a new EU regulation names endocrine disrupting properties as one of the cut-off criteria for the approval of plant protection products, although it currently fails to provide specific science-based measures for the assessment of substances with such properties. Since specific measures are to be presented by the European Commission in 2013 the development of assessment and decision criteria is a key challenge concerning the implementation of this new EU regulation. Proposals of such decision criteria for substances with potential endocrine disrupting properties in human health risk assessment were developed by the German Federal Institute for Risk Assessment (BfR) and discussed at an expert workshop in November 2009. Under consideration of the requirements laid down within the new plant protection product legislation and the scientific discussions during the workshop, a conceptual framework on evaluation of substances for endocrine disrupting properties in a regulatory context is presented in this paper. Central aspects of the framework include assessment of adversity of effects, establishment of a mode/mechanism of action in animals, considerations concerning the relevance of effects to humans and two options for a regulatory decision.

Combined retrospective analysis of 498 rat multi-generation reproductive toxicity studies: on the impact of parameters related to F1 mating and F2 offspring.

The multi-generation reproductive toxicity study (OECD TG 416 and USEPA 870.3800) has been extensively used internationally to assess the adverse effects of substances on reproduction. Recently the necessity of producing a second generation to assess the potential for human health risks has been questioned. The present standardized retrospective analysis of the impact of the second generation on overall study outcome combines earlier analyses and includes 498 rat multi-generation studies representing 438 different tested substances. Detailed assessment of study reports revealed no critical differences in sensitivities between the generations on the basis of a consideration of all endpoints evaluated. This analysis indicates that the second generation mating and offspring will very rarely provide critical information. These findings are consistent with the conclusions of previous retrospective analyses conducted by RIVM, USEPA and PMRA and support adoption of the proposed OECD extended one-generation reproductive toxicity study protocol in regulatory risk assessment testing strategies.