Observations of phase changes in monoolein during high viscous injection.

Serial crystallography of membrane proteins often employs high-viscosity injectors (HVIs) to deliver micrometre-sized crystals to the X-ray beam. Typically, the carrier medium is a lipidic cubic phase (LCP) media, which can also be used to nucleate and grow the crystals. However, despite the fact that the LCP is widely used with HVIs, the potential impact of the injection process on the LCP structure has not been reported and hence is not yet well understood. The self-assembled structure of the LCP can be affected by pressure, dehydration and temperature changes, all of which occur during continuous flow injection. These changes to the LCP structure may in turn impact the results of X-ray diffraction measurements from membrane protein crystals. To investigate the influence of HVIs on the structure of the LCP we conducted a study of the phase changes in monoolein/water and monoolein/buffer mixtures during continuous flow injection, at both atmospheric pressure and under vacuum. The reservoir pressure in the HVI was tracked to determine if there is any correlation with the phase behaviour of the LCP. The results indicated that, even though the reservoir pressure underwent (at times) significant variation, this did not appear to correlate with observed phase changes in the sample stream or correspond to shifts in the LCP lattice parameter. During vacuum injection, there was a three-way coexistence of the gyroid cubic phase, diamond cubic phase and lamellar phase. During injection at atmospheric pressure, the coexistence of a cubic phase and lamellar phase in the monoolein/water mixtures was also observed. The degree to which the lamellar phase is formed was found to be strongly dependent on the co-flowing gas conditions used to stabilize the LCP stream. A combination of laboratory-based optical polarization microscopy and simulation studies was used to investigate these observations.

Control of sheep gastrointestinal nematodes on pasture in the tropical semiarid region of Brazil, using Bioverm® (Duddingtonia flagrans).

We aimed to evaluate a Brazilian commercial formulation of the fungus Duddingtonia flagrans (Bioverm®) for controlling gastrointestinal nematodes in sheep kept on native pasture in the Caatinga biome, in the semiarid region of Brazil. Twenty ewes, aged between 12 and 18 months, were divided into two groups. In the treated group, each animal received 1 g of the Bioverm® product for each 10 kg of live weight, daily, together with commercial feed, for 6 months. In the control group, the animals received feed without Bioverm®. Each group remained throughout the experiment in a 1.2-ha paddock. Monthly counts of eggs per gram (EPG) of feces, fecal cultures, packed cell volume (PCV), weight measurements, and collection of leaf mass from the pasture were performed. There was greater reduction in EPG, greater weight gain, and less infestation by infective larvae in the paddock of the Bioverm® group, compared with the control group (p < 0.05). There was no significant difference (p ≥ 0.05) in the mean PCV percentage between the Bioverm® and control groups. In coprocultures, Haemonchus sp. was the most prevalent helminth. Bioverm® (D. flagrans) was efficient for biological control of sheep gastrointestinal nematodes in the semiarid region of Brazil.

Preferred orientation and its effects on intensity-correlation measurements.

Intensity-correlation measurements allow access to nanostructural information on a range of ordered and disordered materials beyond traditional pair-correlation methods. In real space, this information can be expressed in terms of a pair-angle distribution function (PADF) which encodes three- and four-body distances and angles. To date, correlation-based techniques have not been applied to the analysis of microstructural effects, such as preferred orientation, which are typically investigated by texture analysis. Preferred orientation is regarded as a potential source of error in intensity-correlation experiments and complicates interpretation of the results. Here, the theory of preferred orientation in intensity-correlation techniques is developed, connecting it to the established theory of texture analysis. The preferred-orientation effect is found to scale with the number of crystalline domains in the beam, surpassing the nanostructural signal when the number of domains becomes large. Experimental demonstrations are presented of the orientation-dominant and nanostructure-dominant cases using PADF analysis. The results show that even minor deviations from uniform orientation produce the strongest angular correlation signals when the number of crystalline domains in the beam is large.

Stability, flow alignment and a phase transition of the lipidic cubic phase during continuous flow injection.

Continuous flow injection is a key technology for serial crystallography measurements of protein crystals suspended in the lipidic cubic phase (LCP). To date, there has been little discussion in the literature regarding the impact of the injection process itself on the structure of the lipidic phase. This is despite the fact that the phase of the injection matrix is critical for the flow properties of the stream and potentially for sample stability. Here we report small-angle X-ray scattering measurements of a monoolein:water mixture during continuous delivery using a high viscosity injector. We observe both an alignment and modification of the LCP as a direct result of the injection process. The orientation of the cubic lattice with respect to the beam was estimated based on the anisotropy of the diffraction pattern and does not correspond to a single low order zone axis. The solvent fraction was also observed to impact the stability of the cubic phase during injection. In addition, depending on the distance traveled by the lipid after exiting the needle, the phase is observed to transition from a pure diamond phase (Pn3m) to a mixture containing both gyriod (Ia3d) and lamellar (Lα) phases. Finite element modelling of the observed phase behaviour during injection indicates that the pressure exerted on the lipid stream during extrusion accounts for the variations in the phase composition of the monoolein:water mixture.

Prevalence and isolation of Toxoplasma gondii in goats slaughtered for human consumption in the semi-arid of northeastern Brazil.

The aim of this study was to determine the seroprevalence of anti-Toxoplasma gondii antibodies and factors associated with infection in goats, and to isolate protozoan strains in tissue samples from seropositive goats that were destined for human consumption in the state of Paraíba, northeastern Brazil. Serum samples from 229 slaughtered goats were tested using the indirect fluorescence antibody test (IFAT), with a cutoff point of 1:64. Epidemiological questionnaires were applied to the producers, to acquire information about the sanitary management used in their herds. Tissue samples from the animals were collected during slaughter, in order to perform bioassays in mice. The seroprevalence found was 21.39% (49/229), with antibody titers ranging from 1:64 to 1:32,768. The municipalities of origin, Patos (OR: 3.047; CI: 1.384-6.706) and Sousa (OR: 3.355; CI: 1.536-7.327), were considered to be factors associated with infection by T. gondii. Thirty-eight bioassays were performed in mice, using tissues from seropositive goats, with an isolation rate of 50% (19/38). There was no correlation between isolation rate and antibody titers. Only one mouse died, at 30 days post-infection, which demonstrated that the strains isolated had low virulence towards mice. It was concluded that there is high seroprevalence in goats in northeastern Brazil, as well as a high percentage of viable tissue cysts in slaughtered animals destined for human consumption. These results demonstrate that there is an imminent one health problem relating to toxoplasmosis, especially in the most populous municipalities in the study (Patos and Sousa), which were identified as factors associated with T. gondii infection in goats.

Efficacy of a commercial fungal formulation containing Duddingtonia flagrans (Bioverm®) for controlling bovine gastrointestinal nematodes.

Bioverm® (Duddingtonia flagrans) is a fungal formulation indicated for controlling gastrointestinal nematodes in ruminants and horses, which has recently been authorized for commercialization in Brazil. The objective was to determine the efficiency of Bioverm® against larvae of gastrointestinal nematodes after passage through the gastrointestinal tract of cattle. Twelve animals were used, divided into two groups. In the treated group, a single dose of 1 g of Bioverm® per 10 kg of live weight (containing 105 chlamydospores of D. flagrans) was provided for each animal. Fecal samples were obtained from the animals in each group at 12, 24, 36, 48, 60 and 72 hours after administration. In assay A, 2 g of feces were added to Petri dishes containing 2% agar-water medium. In assay B, coprocultures were performed. In both assays, the peak of larval predation occurred within 48 hours after administration of Bioverm®. In assay A, a significant larval reduction (P < 0.05) was seen at 48 h (88.2%). In assay B, significant reductions (P < 0.05) were seen at 36 h (43.7%) and 48 h (82.3%). Bioverm® showed high predatory capacity after passage through the gastrointestinal tract of cattle and was effective for controlling gastrointestinal nematodes.

Efficacy of a commercial fungal formulation containing Duddingtonia flagrans (Bioverm®) for controlling bovine gastrointestinal nematodes / Eficácia de uma formulação comercial contendo Duddingtonia flagrans (Bioverm®) no controle de nematódeos gastrintestinais de bovinos

Abstract Bioverm® (Duddingtonia flagrans) is a fungal formulation indicated for controlling gastrointestinal nematodes in ruminants and horses, which has recently been authorized for commercialization in Brazil. The objective was to determine the efficiency of Bioverm® against larvae of gastrointestinal nematodes after passage through the gastrointestinal tract of cattle. Twelve animals were used, divided into two groups. In the treated group, a single dose of 1 g of Bioverm® per 10 kg of live weight (containing 105 chlamydospores of D. flagrans) was provided for each animal. Fecal samples were obtained from the animals in each group at 12, 24, 36, 48, 60 and 72 hours after administration. In assay A, 2 g of feces were added to Petri dishes containing 2% agar-water medium. In assay B, coprocultures were performed. In both assays, the peak of larval predation occurred within 48 hours after administration of Bioverm®. In assay A, a significant larval reduction (P < 0.05) was seen at 48 h (88.2%). In assay B, significant reductions (P < 0.05) were seen at 36 h (43.7%) and 48 h (82.3%). Bioverm® showed high predatory capacity after passage through the gastrointestinal tract of cattle and was effective for controlling gastrointestinal nematodes.

Resumo O Bioverm® (Duddingtonia flagrans) é uma formulação fúngica indicada para o controle de nematódeos gastrintestinais de ruminantes e equídeos, recentemente autorizado para a comercialização no Brasil. Objetivou-se determinar a eficiência do Bioverm® contra larvas de nematódeos gastrintestinais após a passagem pelo trato gastrintestinal de bovinos. Foram utilizados doze bovinos divididos em dois grupos. No grupo tratado, foi fornecida, por animal, a dose única de 1g (105 clamidósporos de D. flagrans) do Bioverm® para cada 10 kg de peso vivo. Foram obtidas amostras fecais dos animais de cada grupo a partir de 12, 24, 36, 48, 60 e 72 horas após a administração. No ensaio A, 2g de fezes foram adicionadas em placas de Petri contendo meio ágar-água 2%. No ensaio B, foram realizadas coproculturas. Em ambos os ensaios, o pico de predação larval ocorreu em 48 horas após a administração do Bioverm®. No ensaio A, houve redução larval significativa (P<0,05) em 48h (88,2%). No ensaio B, as reduções significativas (P<0,05) ocorreram em 36h (43,7%) e 48h (82,3%). O Bioverm® apresentou elevada capacidade predatória após a passagem pelo trato gastrintestinal de bovinos, sendo eficaz no controle dos nematódeos gastrintestinais.

Estrategia de orientación psicopedagógica para la prevención del síndrome de Burnout Laboral / Psychopedagogical Strategy for preventing Burnout Syndrome at workplace

RESUMEN Introducción: el síndrome de Burnout consiste en una respuesta prolongada del organismo ante los factores estresantes, emocionales e interpersonales que se presentan en el trabajo e incluye fatiga crónica, ineficacia y negación de lo ocurrido. Se suele presentar en aquellas situaciones laborales en las que los excesivos niveles de exigencia ya se han vuelto un hábito inconsciente e incluso socialmente valorado. Objetivo: elaborar una estrategia de orientación educativa a los profesores desde la función orientadora del psicopedagogo, para la prevención del Síndrome de Burnout Laboral. Métodos: se realizó un estudio descriptivo, prospectivo y cualitativo mediante los métodos teóricos y empíricos, como: histórico-lógico, análisis-síntesis, encuestas, entrevistas, observación, técnica grupal e inventario para la evaluación del Burnout. Resultados: los resultados permitieron precisar las manifestaciones de este síndrome y sus insuficiencias. En la muestra estudiada se comprobó que de 19 profesionales se manifiestan en 15 (79%), en alguna medida indicadores de esta afectación, como síntomas de cansancio emocional y la baja realización personal. Conclusiones: la estrategia de orientación aplicada contribuye con la preparación del psicopedagogo en la prevención del Síndrome de Burnout Laboral y perfecciona el proceso docente educativo, proporciona mayor estabilidad psicológica y favorece la unidad de influencias educativas.

ABSTRACT Introduction: Burnout syndrome is provoqued by a prolonged response of the organism to stress, emotional and interpersonal factors such as chronic fatigue, inefficiency and refutation. It is usually present in those workers with ridiculous levels of tasks have already become an unconscious and social habit. Objective: to elaborate an educational guidance strategy for teachers, according to their function of Psychopedagogue for Burnout syndrome prevention. Methods: a descriptive, prospective and qualitative study was carried out using theoretical and empirical methods such as: historical-logical, analysis-synthesis, surveys, interviews, observation, group technique and Burnout syndrome records. Results: there were insufficiencies. From the total sample of 19 professionals, some indicators revealed 15 (79%) of participants with emotional fatigue and low self-realization highlighted symptoms. Conclusions: the strategy contributes to prepare the psychopedagogue on Burnout syndrome prevention and improves the educational process. It also provides greater psychological stability and favors educational influences unity.

Biodepuration of domestic sewage, textile effluents and acid mine drainage using starch-based xerogel from recycled potato peels.

Water is a finite resource. Its safety and cleanliness are highly important to meet current and future human needs. Compared to other resources, water represents a main factor to achieve development in several areas and leads to economic progress of a nation. However, in recent years, the excessive demographic and industrial growth has exacerbated water contamination. In this study, the biodepuration process of domestic sewage (DS), textile effluents (TE) and acid mine drainage (AMD) is conducted using starch-based xerogel from potato (Solanum tuberosum) peels. Results showed that the treatment is effective to achieve the reduction of 5-day biochemical oxygen demand. The most important result was the achievement of heavy metals removal for the three components. Firstly, there was a reduction of barium, zinc, and cadmium (91, 60 and 46%, respectively) for raw AMD. Secondly, there was a reduction in the levels of zinc, aluminum, and barium (89, 86 and 64%, respectively) for TE biodepuration. Finally, results showed a reduction in zinc, iron and cadmium levels (81, 78 and 57%, respectively) for DS biodepuration.

GBA2 Mutations Cause a Marinesco-Sjögren-Like Syndrome: Genetic and Biochemical Studies.

BACKGROUND: With the advent new sequencing technologies, we now have the tools to understand the phenotypic diversity and the common occurrence of phenocopies. We used these techniques to investigate two Norwegian families with an autosomal recessive cerebellar ataxia with cataracts and mental retardation. METHODS AND RESULTS: Single nucleotide polymorphism (SNP) chip analysis followed by Exome sequencing identified a 2 bp homozygous deletion in GBA2 in both families, c.1528_1529del [p.Met510Valfs*17]. Furthermore, we report the biochemical characterization of GBA2 in these patients. Our studies show that a reduced activity of GBA2 is sufficient to elevate the levels of glucosylceramide to similar levels as seen in Gaucher disease. Furthermore, leucocytes seem to be the proper enzyme source for in vitro analysis of GBA2 activity. CONCLUSIONS: We report GBA2 mutations causing a Marinesco-Sjögren-like syndrome in two Norwegian families. One of the families was originally diagnosed with Marinesco-Sjögren syndrome based on an autosomal recessive cerebellar ataxia with cataracts and mental retardation. Our findings highlight the phenotypic variability associated with GBA2 mutations, and suggest that patients with Marinesco-Sjögren-like syndromes should be tested for mutations in this gene.

Disentangling genetic and epigenetic determinants of ultrafast adaptation.

A major rationale for the advocacy of epigenetically mediated adaptive responses is that they facilitate faster adaptation to environmental challenges. This motivated us to develop a theoretical-experimental framework for disclosing the presence of such adaptation-speeding mechanisms in an experimental evolution setting circumventing the need for pursuing costly mutation-accumulation experiments. To this end, we exposed clonal populations of budding yeast to a whole range of stressors. By growth phenotyping, we found that almost complete adaptation to arsenic emerged after a few mitotic cell divisions without involving any phenotypic plasticity. Causative mutations were identified by deep sequencing of the arsenic-adapted populations and reconstructed for validation. Mutation effects on growth phenotypes, and the associated mutational target sizes were quantified and embedded in data-driven individual-based evolutionary population models. We found that the experimentally observed homogeneity of adaptation speed and heterogeneity of molecular solutions could only be accounted for if the mutation rate had been near estimates of the basal mutation rate. The ultrafast adaptation could be fully explained by extensive positive pleiotropy such that all beneficial mutations dramatically enhanced multiple fitness components in concert. As our approach can be exploited across a range of model organisms exposed to a variety of environmental challenges, it may be used for determining the importance of epigenetic adaptation-speeding mechanisms in general.

Alternativas para lograr la revascularización arterial total usando una o ambas arterias mamarias y el remanente distal de una de ellas como únicos injertos / Alternatives to achieve total arterial revascularization using one or both mammary arteries and distal remnant of them as unique grafts

Introducción: Toda cirugía de revascularización miocárdica debe estar dirigida a lograr que el paciente permanezca el mayor tiempo posible sin necesidad de una nueva revascularización. La revascularización arterial total contribuye a alcanzar este propósito. Objetivo: Demostrar que ambas arterias mamarias pueden ser utilizadas, en varias configuraciones, para lograr la revascularización arterial total. Métodos: Se incluyeron 35 pacientes, 29 de género masculino y 6 del femenino, con edad promedio de 54,2 años, a quienes se les revascularizaron una o ambas arterias mamarias, en el período entre enero de 2012 y septiembre de 2014, para un seguimiento medio de 13,1 meses. El valor del Euroscore aditivo fue de 0 para los hombres y 1 para las mujeres. Resultados: Se describieron siete configuraciones para lograr la revascularización arterial total, usando una o ambas arterias mamarias y el remanente distal de una de las arterias mamarias como únicos injertos. No hubo fallecimientos ni complicaciones mayores en los pacientes revascularizados. Seis meses después todos permanecían asintomáticos y sin nuevos signos de isquemia en los territorios revascularizados. Conclusiones: La revascularización arterial total, utilizando una o ambas arterias mamarias y el remanente distal de una de ellas es posible y arroja buenos resultados en pacientes seleccionados.

Introduction: The aim of all coronary artery bypass grafts should be to allow patients to stay as long as possible without the need to repeat revascularization. Total arterial revascularization contributes to achieving these purposes. Objective: To demonstrate that both mammary arteries can be used in various configurations to achieve total arterial revascularization. Methods: 35 patients were included, 29 male and 6 female, with age average of 54.2 years, who were revascularized with one or both mammary arteries in the period between January 2012 and September 2014, with a mean follow-up time of 13.1 months. The additive EuroSCORE was 0 for men and 1 for women. Results: Seven configurations are described to achieve total arterial revascularization using one or both mammary arteries and the distal remnant of a mammary arteries as single grafts. There were no deaths or major complications in patients revascularized. Six months later, all were asymptomatic with no new signs of ischemia in revascularized territories. Conclusions: Total arterial revascularization using one or both mammary arteries and distal remaining one is possible and gives good results in selected patients.

[Pituitary tumors: 10 years of experience]. / Adenomas hipofisarios: análisis de la casuística de 10 años.

INTRODUCTION: Pituitary tumors comprise a quarter of the intracranial neoplasms and the adenomas represent the highest percentage of them. They are benign, but they can be invasive and they have an impact in morbidity and mortality. OBJECTIVE: To analyze the clinical features of pituitary adenomas tumors, and create a computerized registry to improve strategies for diagnosis and monitoring of these patients. METHODS: A retrospective, descriptive analysis of 102 medical records of a total of 191 patients who attended the endocrinology service from 2003 to 2014 .The variables were analyzed by the corresponding descriptive statistics. The SPSS 11.5 was used. RESULTS: 63% were women, and (74.4%) had age from 25-60 years. 54% were nonfunctioning adenomas, 28% to prolactinomas, 11.8% and 6.2% somatotropinomas corticotropinomas. In all types the female group was predominant, except nonfunctioning. Of the nonfunctioning adenomas 79% were macroadenomas, prolactinomas 52% were microadenomas and surgery was performed in 37% of them. The somatotropinomas, 80% were macroadenomas, 80% of patients and 40% radiotherapy was operated. In corticotropinomas, both micro and macroadenomas were submitted by the same percentage (50%) and all of them underwent surgery. CONCLUSION: The findings are consistent with the literature in terms of frequency and age of onset of adenomas. We suggest the development of a registry to optimize the monitoring and treatment of patients, research and scientific publication.

Introducción: Los tumores hipofisarios comprenden la cuarta parte de las neoplasias intracraneales y los adenomas son el mayor porcentaje de ellos. Son de naturaleza benigna, pero pueden ser invasivos y producir impacto en la morbi-mortalidad. Objetivo: analizar las características clínicas de los pacientes con diagnóstico de adenomas hipofisarios y crear un registro computarizado para mejorar las estrategias de diagnóstico y seguimiento de estos pacientes. Metodología: Se realizó un análisis retrospectivo, descriptivo, de 102 historias clínicas sobre un total de 191 pacientes que concurrieron al servicio de endocrinología desde el año 2003 al 2014 . Las variables fueron analizadas mediante las correspondientes estadísticas descriptivas. Se utilizó el programa SPSS 11.5. Resultados: El 63% fueron mujeres, y el mayor porcentaje etario entre 25-60 años (74.4%). El 54% correspondió a adenomas no funcionantes, el 28% a prolactinomas, el 11.8% somatotropinomas y el 6.2% corticotropinomas. En todos los tipos el grupo femenino fue mayoritario, excepto en los no funcionantes. De los adenomas no funcionantes el 79% fueron macroadenomas, los prolactinomas el 52% fueron microadenomas y se realizó cirugía en el 37% de ellos. Los somatotropinomas, en el 80% fueron macroadenomas, se operó el 80% de los pacientes y radioterapia al 40%. En los corticotropinomas, tanto micro como macroadenomas se presentaron en igual porcentaje (50%) y todos ellos recibieron tratamiento quirúrgico. Conclusión: Los hallazgos coinciden con la literatura en cuanto a frecuencia y edad de presentación de los adenomas. Consideramos valioso la elaboración de un registro que nos permita optimizar el seguimiento y tratamiento de los pacientes, la investigación y divulgación científica.

Impacto económico del precondicionamiento isquémico remoto en la revascularización miocárdica / Economic impact of remote ischemic preconditioning on myocardial revascularization

Introducción: condicionar el corazón para mejorar sus capacidades cardioprotectoras endógenas con el uso de isquemias breves a distancia proporciona un novedoso abordaje potencial a la protección miocárdica durante la cirugía cardiaca. Objetivo: identificar el impacto económico del precondicionamiento isquémico a distancia en los pacientes revascularizados quirúrgicamente. Métodos: se realizó un estudio longitudinal prospectivo experimental en dos grupos de 100 personas, a los que se les realizó revascularización por injerto de la arteria coronaria. Se procedió a colocar un torniquete en el brazo no dominante en quienes se incluyeron en el grupo de estudio prueba, alternando tres insuflaciones con tres desinsuflaciones con una presión de 200 mmHg, manteniéndola por espacio de cinco minutos cada una, este proceder se realizó previo, durante y después del evento isquémico mayor que se corresponde con el pinzamiento de la arteria coronaria. Resultados: se logró una importante disminución del consumo de drogas inotrópicas, vasoactivas y de otros medicamentos ahorrándose una importante suma disminuyendo los costos hospitalarios. Comprobándose además, la disminución en la incidencia de arritmias ventriculares letales, bajo gasto cardiaco fatal y de muerte postoperatoria, en todos los casos muy por debajo de la predicción previamente realizada para estas complicaciones. Conclusiones: el precondicionamiento isquémico a distancia puede ser una importante herramienta a tener en cuenta en la protección antisquémica de la revascularización miocárdica que puede disminuir la morbimortalidad y los costos hospitalarios.

Background: to condition the heart to improve its endogenous cardioprotective capacity using brief remote ischemia provides a novel potential approach to myocardial protection during cardiac surgery. Objective: to identify the economic impact of remote ischemic preconditioning in surgically revascularized patients. Methods: an experimental prospective longitudinal study was conducted in two groups of 100 people who underwent revascularization by coronary artery graft. A tourniquet was placed on the non-dominant arm in those who were included in the test study group, alternating three insufflations with three desinsufflations with a pressure of 200 mmHg, each one being maintained for five minutes. This procedure was performed prior to, during and after the greater ischemic event that corresponds to the pinching of the coronary artery. Results: an important decrease of the consumption of inotropic, vasoactive and other drugs was achieved, saving an important sum, decreasing hospital costs, and also proving a reduction in the incidence of lethal ventricular arrhythmias, low cardiac output and postoperative death which were, in all cases, below the prediction previously made for these complications. Conclusions: remote ischemic preconditioning can be an important tool to be considered in the antischemic protection of myocardial revascularization that can diminish morbimortality and hospital costs.

Exome sequencing and genetic testing for MODY.

CONTEXT: Genetic testing for monogenic diabetes is important for patient care. Given the extensive genetic and clinical heterogeneity of diabetes, exome sequencing might provide additional diagnostic potential when standard Sanger sequencing-based diagnostics is inconclusive. OBJECTIVE: The aim of the study was to examine the performance of exome sequencing for a molecular diagnosis of MODY in patients who have undergone conventional diagnostic sequencing of candidate genes with negative results. RESEARCH DESIGN AND METHODS: We performed exome enrichment followed by high-throughput sequencing in nine patients with suspected MODY. They were Sanger sequencing-negative for mutations in the HNF1A, HNF4A, GCK, HNF1B and INS genes. We excluded common, non-coding and synonymous gene variants, and performed in-depth analysis on filtered sequence variants in a pre-defined set of 111 genes implicated in glucose metabolism. RESULTS: On average, we obtained 45 X median coverage of the entire targeted exome and found 199 rare coding variants per individual. We identified 0-4 rare non-synonymous and nonsense variants per individual in our a priori list of 111 candidate genes. Three of the variants were considered pathogenic (in ABCC8, HNF4A and PPARG, respectively), thus exome sequencing led to a genetic diagnosis in at least three of the nine patients. Approximately 91% of known heterozygous SNPs in the target exomes were detected, but we also found low coverage in some key diabetes genes using our current exome sequencing approach. Novel variants in the genes ARAP1, GLIS3, MADD, NOTCH2 and WFS1 need further investigation to reveal their possible role in diabetes. CONCLUSION: Our results demonstrate that exome sequencing can improve molecular diagnostics of MODY when used as a complement to Sanger sequencing. However, improvements will be needed, especially concerning coverage, before the full potential of exome sequencing can be realized.

Using electronic patient records to discover disease correlations and stratify patient cohorts.

Electronic patient records remain a rather unexplored, but potentially rich data source for discovering correlations between diseases. We describe a general approach for gathering phenotypic descriptions of patients from medical records in a systematic and non-cohort dependent manner. By extracting phenotype information from the free-text in such records we demonstrate that we can extend the information contained in the structured record data, and use it for producing fine-grained patient stratification and disease co-occurrence statistics. The approach uses a dictionary based on the International Classification of Disease ontology and is therefore in principle language independent. As a use case we show how records from a Danish psychiatric hospital lead to the identification of disease correlations, which subsequently can be mapped to systems biology frameworks.

ChemProt: a disease chemical biology database.

Systems pharmacology is an emergent area that studies drug action across multiple scales of complexity, from molecular and cellular to tissue and organism levels. There is a critical need to develop network-based approaches to integrate the growing body of chemical biology knowledge with network biology. Here, we report ChemProt, a disease chemical biology database, which is based on a compilation of multiple chemical-protein annotation resources, as well as disease-associated protein-protein interactions (PPIs). We assembled more than 700,000 unique chemicals with biological annotation for 30,578 proteins. We gathered over 2-million chemical-protein interactions, which were integrated in a quality scored human PPI network of 428,429 interactions. The PPI network layer allows for studying disease and tissue specificity through each protein complex. ChemProt can assist in the in silico evaluation of environmental chemicals, natural products and approved drugs, as well as the selection of new compounds based on their activity profile against most known biological targets, including those related to adverse drug events. Results from the disease chemical biology database associate citalopram, an antidepressant, with osteogenesis imperfect and leukemia and bisphenol A, an endocrine disruptor, with certain types of cancer, respectively. The server can be accessed at http://www.cbs.dtu.dk/services/ChemProt/.

Dissecting spatio-temporal protein networks driving human heart development and related disorders.

Aberrant organ development is associated with a wide spectrum of disorders, from schizophrenia to congenital heart disease, but systems-level insight into the underlying processes is very limited. Using heart morphogenesis as general model for dissecting the functional architecture of organ development, we combined detailed phenotype information from deleterious mutations in 255 genes with high-confidence experimental interactome data, and coupled the results to thorough experimental validation. Hereby, we made the first systematic analysis of spatio-temporal protein networks driving many stages of a developing organ identifying several novel signaling modules. Our results show that organ development relies on surprisingly few, extensively recycled, protein modules that integrate into complex higher-order networks. This design allows the formation of a complicated organ using simple building blocks, and suggests how mutations in the same genes can lead to diverse phenotypes. We observe a striking temporal correlation between organ complexity and the number of discrete functional modules coordinating morphogenesis. Our analysis elucidates the organization and composition of spatio-temporal protein networks that drive the formation of organs, which in the future may lay the foundation of novel approaches in treatments, diagnostics, and regenerative medicine.

Deciphering diseases and biological targets for environmental chemicals using toxicogenomics networks.

Exposure to environmental chemicals and drugs may have a negative effect on human health. A better understanding of the molecular mechanism of such compounds is needed to determine the risk. We present a high confidence human protein-protein association network built upon the integration of chemical toxicology and systems biology. This computational systems chemical biology model reveals uncharacterized connections between compounds and diseases, thus predicting which compounds may be risk factors for human health. Additionally, the network can be used to identify unexpected potential associations between chemicals and proteins. Examples are shown for chemicals associated with breast cancer, lung cancer and necrosis, and potential protein targets for di-ethylhexyl-phthalate, 2,3,7,8-tetrachlorodibenzo-p-dioxin, pirinixic acid and permethrine. The chemical-protein associations are supported through recent published studies, which illustrate the power of our approach that integrates toxicogenomics data with other data types.