RAG1/2 induziert genomische Insertionen durch die Mobilisierung von DNA in RAG1/2-unabhängige Brüche

The RAG recombinase (RAG1/2) plays an essential role in adaptive immunity by mediating V(D)J recombination in developing lymphocytes. In contrast, aberrant RAG1/2 activity promotes lymphocyte malignancies by causing chromosomal translocations and DNA deletions at cancer genes. In addition, RAG1/2 can induce aberrant DNA insertions by transposition and trans-V(D)J recombination, but only few putative such events have been documented in vivo. Moreover, those observed in cancer display characteristics that are not compatible with either DNA transposition or trans-V(D)J recombination. Hence, how RAG1/2 causes genomic DNA insertions is still largely unknown. In this study, I use translocation capture sequencing (TC-Seq) and insertion capture sequencing (IC-Seq) to analyze chromosomal rearrangements in primary murine developing B cells. I identify aberrant RAG1/2-dependent DNA deletions at immunoglobulin genes, whose products are re-inserted at DNA breaks generated by the I-SceI endonuclease on a heterologous chromosome. The existence of similar insertions in human cancer indicates that RAG1/2 also mobilizes genomic DNA into independent physiologic breaks in vivo. Thus, my findings reveal a novel pathway through which RAG1/2 causes DNA insertions independent of DNA transposition and trans-V(D)J recombination. Importantly, this pathway has the potential to destabilize the lymphocyte genome by causing aberrant signal-end, hybrid-end and coding-end insertions and shares features with reported oncogenic DNA insertions

The Excess Costs of Depression and the Influence of Sociodemographic and Socioeconomic Factors: Results from the German Health Interview and Examination Survey for Adults (DEGS)

Introduction The aim of this study was to estimate excess costs of depression in Germany and to examine the influence of sociodemographic and socioeconomic determinants. Methods Annual excess costs of depression per patient were estimated for the year 2019 by comparing survey data of individuals with and without self-reported medically diagnosed depression, representative for the German population aged 18–79 years. Differences between individuals with depression (n = 223) and without depression (n = 4540) were adjusted using entropy balancing. Excess costs were estimated using generalized linear model regression with a gamma distribution and log-link function. We estimated direct (inpatient, outpatient, medication) and indirect (sick leave, early retirement) excess costs. Subgroup analyses by social determinants were conducted for sex, age, socioeconomic status, first-generation or second-generation migrants, partnership, and social support. Results Total annual excess costs of depression amounted to €5047 (95% confidence interval [CI] 3214–6880) per patient. Indirect excess costs amounted to €2835 (1566–4103) and were higher than direct excess costs (€2212 [1083–3341]). Outpatient (€498), inpatient (€1345), early retirement (€1686), and sick leave (€1149) excess costs were statistically significant, while medication (€370) excess costs were not. Regarding social determinants, total excess costs were highest in the younger age groups (€7955 for 18–29-year-olds, €9560 for 30–44-year-olds), whereas total excess costs were lowest for the oldest age group (€2168 for 65+) and first-generation or second-generation migrants (€1820). Conclusions Depression was associated with high excess costs that varied by social determinants. Considerable differences between the socioeconomic and sociodemographic subgroups need further clarification as they point to specific treatment barriers as well as varying treatment needs.Peer Reviewe

Cytotoxicity screening of 23 engineered nanomaterials using a test matrix of ten cell lines and three different assays

<p>Abstract</p> <p>Background</p> <p>Engineered nanomaterials display unique properties that may have impact on human health, and thus require a reliable evaluation of their potential toxicity. Here, we performed a standardized <it>in vitro </it>screening of 23 engineered nanomaterials. We thoroughly characterized the physicochemical properties of the nanomaterials and adapted three classical <it>in vitro </it>toxicity assays to eliminate nanomaterial interference. Nanomaterial toxicity was assessed in ten representative cell lines.</p> <p>Results</p> <p>Six nanomaterials induced oxidative cell stress while only a single nanomaterial reduced cellular metabolic activity and none of the particles affected cell viability. Results from heterogeneous and chemically identical particles suggested that surface chemistry, surface coating and chemical composition are likely determinants of nanomaterial toxicity. Individual cell lines differed significantly in their response, dependent on the particle type and the toxicity endpoint measured.</p> <p>Conclusion</p> <p><it>In vitro </it>toxicity of the analyzed engineered nanomaterials cannot be attributed to a defined physicochemical property. Therefore, the accurate identification of nanomaterial cytotoxicity requires a matrix based on a set of sensitive cell lines and <it>in vitro </it>assays measuring different cytotoxicity endpoints.</p

Evaluación del proceso productivo de Mermeladas en la Asociación ASOPRUV.

The research project is focus on evaluate and analyze the jam production and elaboration process in the ASOPRUV association, which does not have a technical productive organization and whose processes, methods and time are not standardized; in addition, the distribution of the machinery in the plant is not established in a suitable way, as a result which makes the process of production has low levels, and also the factory permanence as competitive factory in the Ecuadorian market is in risk. The evaluation and analysis was realized with the previous information was generated with the actual situation by applying a fiel research, the producer involves the identification of the products and its elaboration process, for this a flow chart was elaborated that allows the study of time and movements determining the actual productivity. The planned activities were reached through the determination of errors that affect the process by the use of stablished methods like cause - effect diagrams. The design of the Plant and the distribution of the equipment are the principal components for the balance of the lines of production, this the proposal of redistribution of machinery to have a better productivity. Other method that was applied was the inverse diagram to establish the solution of the found errors in the process of jam elaboration, finally an improvement proposal was proposed, for which a layout of the company was made with the old model and improved model that is reflected with the greatest optimization of time and movements, in addition a coding machine of dates of elaboration and expiration was acquired, for the jam containers that are made of glass, which made better the quality and control of the products, opening the possibility of use international regulations in the future.El proyecto de investigación se enfoca en evaluar y analizar el proceso productivo de elaboración de mermeladas en la asociación ASOPRUV, la cual carece de una organización técnicamente productiva y sin estandarizar los procesos, métodos y tiempos; además la distribución de la maquinaria en la planta está establecida inadecuadamente, lo que hace que el proceso de producción experimente niveles bajos de productividad y ponga en riesgo su permanencia como empresa competitiva en el mercado ecuatoriano. La evaluación y el análisis se realizó con el previo levantamiento de información que se generó bajo el estado actual de la empresa aplicando una investigación de campo, procediendo con la identificación de los productos fabricados y su proceso para lo cual se elaboró un diagrama de flujo de procesos que permita, el estudio de tiempos y movimientos determinando así la productividad actual. Las actividades planteadas se lograron cumplir mediante la determinación de fallas que inciden en el proceso, bajo métodos establecidos como diagramas de causa-efecto. El diseño de la planta y la distribución de equipos son los componentes principales para el balanceo de las líneas de producción, lo cual se propone la redistribución de máquinas en planta para tener una mejor productividad. Otro método que se realizó, es el diagrama inverso para establecer las soluciones de los errores encontrados en el proceso de elaboración de mermelada, por último se planteó la propuesta de mejora, para lo cual se realizó un Layout de la empresa con el modelo antiguo y modelo mejorado que se refleja con la mayor optimización de tiempos y movimientos, adicionalmente se adquirió una máquina codificadora de fechas de elaboración y caducidad para los envases de la mermelada que son de vidrio, lo cual mejoró la calidad y el control de los productos, abriéndose la posibilidad de ejercer normativas internacionales en el futuroUniversidad Técnica de Cotopax

A Chemical Proteomics Approach to Phosphatidylinositol 3-Kinase Signaling in Macrophages

Prior work using lipid-based affinity matrices has been done to investigate distinct sets of lipid-binding proteins, and one series of experiments has proven successful in mammalian cells for the proteome-wide identification of lipid-binding proteins. However, most lipid-based proteomics screens require scaled up sample preparation, are often composed of multiple cell types, and are not adapted for simultaneous signal transduction studies. Herein we provide a chemical proteomics strategy that uses cleavable lipid "baits" with broad applicability to diverse biological samples. The novel baits were designed to avoid preparative steps to allow functional proteomics studies when the biological source is a limiting factor. Validation of the chemical baits was first confirmed by the selective isolation of several known endogenous phosphatidylinositol 3-kinase signaling proteins using primary bone marrow-derived macrophages. The use of this technique for cellular proteomics and MS/MS analysis was then demonstrated by the identification of known and potential novel lipid-binding proteins that was confirmed in vitro for several proteins by direct lipid-protein interactions. Further to the identification, the method is also compatible with subsequent signal transduction studies, notably for protein kinase profiling of the isolated lipid-bound protein complexes. Taken together, this integration of minimal scale proteomics, lipid chemistry, and activity-based readouts provides a significant advancement in the ability to identify and study the lipid proteome of single, relevant cell types

The p110 delta structure: mechanisms for selectivity and potency of new PI(3)K inhibitors.

Deregulation of the phosphoinositide-3-OH kinase (PI(3)K) pathway has been implicated in numerous pathologies including cancer, diabetes, thrombosis, rheumatoid arthritis and asthma. Recently, small-molecule and ATP-competitive PI(3)K inhibitors with a wide range of selectivities have entered clinical development. In order to understand the mechanisms underlying the isoform selectivity of these inhibitors, we developed a new expression strategy that enabled us to determine to our knowledge the first crystal structure of the catalytic subunit of the class IA PI(3)K p110 delta. Structures of this enzyme in complex with a broad panel of isoform- and pan-selective class I PI(3)K inhibitors reveal that selectivity toward p110 delta can be achieved by exploiting its conformational flexibility and the sequence diversity of active site residues that do not contact ATP. We have used these observations to rationalize and synthesize highly selective inhibitors for p110 delta with greatly improved potencies

A randomization-based causal inference framework for uncovering environmental exposure effects on human gut microbiota

Statistical analysis of microbial genomic data within epidemiological cohort studies holds the promise to assess the influence of environmental exposures on both the host and the host-associated microbiome. However, the observational character of prospective cohort data and the intricate characteristics of microbiome data make it challenging to discover causal associations between environment and microbiome. Here, we introduce a causal inference framework based on the Rubin Causal Model that can help scientists to investigate such environment-host microbiome relationships, to capitalize on existing, possibly powerful, test statistics, and test plausible sharp null hypotheses. Using data from the German KORA cohort study, we illustrate our framework by designing two hypothetical randomized experiments with interventions of (i) air pollution reduction and (ii) smoking prevention. We study the effects of these interventions on the human gut microbiome by testing shifts in microbial diversity, changes in individual microbial abundances, and microbial network wiring between groups of matched subjects via randomization-based inference. In the smoking prevention scenario, we identify a small interconnected group of taxa worth further scrutiny, including Christensenellaceae and Ruminococcaceae genera, that have been previously associated with blood metabolite changes. These findings demonstrate that our framework may uncover potentially causal links between environmental exposure and the gut microbiome from observational data. We anticipate the present statistical framework to be a good starting point for further discoveries on the role of the gut microbiome in environmental health

Mitigation of interfacial dielectric loss in aluminum-on-silicon superconducting qubits

We demonstrate aluminum-on-silicon planar transmon qubits with time-averaged ${T_1}$ energy relaxation times of up to ${270\,\mu s}$, corresponding to Q = 5 million, and a highest observed value of ${501\,\mu s}$. We use materials analysis techniques and numerical simulations to investigate the dominant sources of energy loss, and devise and demonstrate a strategy towards mitigating them. The mitigation of loss is achieved by reducing the presence of oxide, a known host of defects, near the substrate-metal interface, by growing aluminum films thicker than 300 nm. A loss analysis of coplanar-waveguide resonators shows that the improvement is owing to a reduction of dielectric loss due to two-level system defects. We perform time-of-flight secondary ion mass spectrometry and observe a reduced presence of oxygen at the substrate-metal interface for the thicker films. The correlation between the enhanced performance and the film thickness is due to the tendency of aluminum to grow in columnar structures of parallel grain boundaries, where the size of the grain depends on the film thickness: transmission electron microscopy imaging shows that the thicker film has larger grains and consequently fewer grain boundaries containing oxide near this interface. These conclusions are supported by numerical simulations of the different loss contributions in the device.Comment: 13 pages, 11 figures, 2 table