Standing sedation management of a domesticated reindeer for third eyelid removal.

Background: Reindeer are becoming popular animals within petting farms. Few case reports describe the sedation of domesticated reindeer, but none describe the use of ocular local anesthetic blocks in this species. Case Description: A 9-year-old, female, Svalbard reindeer (Rangifer tarandus platyrhynchus) presenting for removal of a squamous cell carcinoma involving the third eyelid. Standing sedation was performed using initial boluses of medetomidine and butorphanol via intramuscular injection before catheter placement and maintenance with a variable rate infusion of medetomidine. Supraorbital, auriculopalpebral, infratrochlear blocks and local infiltration of the base of the third eyelid were performed using mepivacaine. Following the surgical removal of the third eyelid, atipamazole was administered intramuscularly to antagonize the effects of medetomidine. The patient recovered without complications. Conclusion: Medetomidine-butorphanol in combination with local anesthetic blocks provided a sufficient plane of sedation and analgesia for extra ocular surgery in a domesticated reindeer.

Prevalence and outcome of lens capsule disruption in routine canine cataract surgery: A retrospective study of 520 eyes (2012-2019).

OBJECTIVE: To investigate the prevalence and surgical outcome of lens capsule disruption (LCD) in dogs undergoing cataract removal. ANIMALS STUDIED: Medical records of 924 eyes undergoing phacoemulsification were analyzed retrospectively. PROCEDURES: Routine cataract surgeries with or without LCD were included. Any LCD other than routine anterior capsulorhexis was defined as LCD and classified according to location and etiology. Odds ratios (OR) were calculated for maintaining vision, implantation of an artificial intraocular lens (IOL), and enucleation. RESULTS: In total, 520 eyes were included. A LCD occurred in 145 eyes (27.8%; 145/520) and affected the posterior (85.5%; 124/145), anterior (6.2%; 9/145), and equatorial lens capsule (4.8%; 7/145) and at multiple locations (3.4%; 5/145). The etiology of the LCD was spontaneous preoperative in 41 eyes (28.3%; 41/145), accidental intraoperative in 57 eyes (39.3%; 57/145), and planned in 47 eyes (32.4%; 47/145). Disruption did not increase the odds of enucleation (OR = 1.48, 95% confidence interval [CI] 0.56-3.67; p = .36). The presence of LCD significantly increased the risk of losing vision 1 year post-operatively (OR = 8.17, 95% CI 1.41-84.93; p = .007) associated with retinal detachment. However, this was not present at 2 years follow-up or in PCCC cases at any time point. An IOL was implanted in 108 eyes (108/145; 75.2%) with LCD and in 45/47 (95.7%) eyes with a PCCC. CONCLUSION: Increased surgeon awareness of possible intraoperative, accidental LCDs is important, as LCDs were relatively common and associated with increased odds for vision loss after 1 year in the present study. A prospective study investigating the causes of intraoperative, accidental LCD is warranted.

The PAX5-JAK2 translocation acts as dual-hit mutation that promotes aggressive B-cell leukemia via nuclear STAT5 activation.

While PAX5 is an important tumor suppressor gene in B-cell acute lymphoblastic leukemia (B-ALL), it is also involved in oncogenic translocations coding for diverse PAX5 fusion proteins. PAX5-JAK2 encodes a protein consisting of the PAX5 DNA-binding region fused to the constitutively active JAK2 kinase domain. Here, we studied the oncogenic function of the PAX5-JAK2 fusion protein in a mouse model expressing it from the endogenous Pax5 locus, resulting in inactivation of one of the two Pax5 alleles. Pax5Jak2/+ mice rapidly developed an aggressive B-ALL in the absence of another cooperating exogenous gene mutation. The DNA-binding function and kinase activity of Pax5-Jak2 as well as IL-7 signaling contributed to leukemia development. Interestingly, all Pax5Jak2/+ tumors lost the remaining wild-type Pax5 allele, allowing efficient DNA-binding of Pax5-Jak2. While we could not find evidence for a nuclear role of Pax5-Jak2 as an epigenetic regulator, high levels of active phosphorylated STAT5 and increased expression of STAT5 target genes were seen in Pax5Jak2/+ B-ALL tumors, implying that nuclear Pax5-Jak2 phosphorylates STAT5. Together, these data reveal Pax5-Jak2 as an important nuclear driver of leukemogenesis by maintaining phosphorylated STAT5 levels in the nucleus.

Influence of the storage period on the nutritional and microbiological value of sun-dried brewer's grains / Influencia del período de almacenamiento sobre el valor nutricional y microbiológico de los granos de cervecería secados al sol / Influência do período de estocagem sobre o valor nutricional e microbiológico de grãos de cervejaria secos ao sol

Abstract Background: Brewer's grains, a by-product of the brewery industry, can be included in the diet of ruminants. However, its high humidity makes it difficult to store and preserve. Objective: To evaluate the efficiency of sun dehydration of wet brewer's grains (WBG) and the effect of storage period on its nutritional and microbiological quality. Methods: A completely randomized experimental design was used to evaluate WBG dehydration efficiency, with treatments corresponding to 0, 1, 2, 4, 6, 8, 10, 12, 14 and 16 hours of sun exposure. A second experiment was carried out using also a completely randomizeddesign to evaluated the effect of storage with the following treatments: 0, 10, 20, 30, 60, 90, 120, 150 and 180 days of storage of the dry by-product. Results: Dry matter (DM) content linearly increased with dehydration period. The chemical composition of the dried brewer's grains had no effect as a function of storage period. Indigestible protein (C fraction) increased linearly but did not compromise the cumulative gas production and the in vitro digestibility of DM and protein. Storage time had no effect on fungus population. The maximum aflatoxin value was 45.5 μg/kg, and remained within acceptable limits for bovine feed. Conclusion: Dehydration of WBG in the sun is efficient to guarantee conservation and makes it possible to store the by- product. The storage of the dry by-product for 180 days does not compromise its nutritional or microbiological quality.

Resumen Antecedentes: Los granos de cervecería son un subproducto de la industria cervecera que puede ser incluido en la dieta de rumiantes; sin embargo, su alta húmedad dificulta el almacenamiento y conservación de ese producto. Objetivo: Evaluar la eficiencia de la deshidratación al sol de los granos húmedos de cervecería (WBG) y el efecto del período de almacenamiento sobre su calidad nutricional y microbiológica. Métodos: Para evaluar la eficiencia de la deshidratación de los WBG se utilizó un diseño experimental completamente al azar, con tiempos de tratamiento de 0, 1, 2, 4, 6, 8, 10, 12, 14, y 16 horas de exposición al sol. En un segundo experimento, también con diseño experimental completamente al azar, se evaluó el efecto del almacenamiento comparando los siguientes tratamientos: 0, 10, 20, 30, 60, 90, 120, 150 y 180 días de almacenamiento del subproducto seco. Resultados: La materia seca (DM) del WBG presentó un efecto lineal creciente con el proceso de deshidratación. La composición química de los granos secos de cervecería no tuvo efecto en función de los tiempos de almacenamiento. La proteína no digestible (fracción C) aumentó linealmente; sin embargo, no comprometió la producción acumulativa de gas y la digestibilidad in vitro de la DM y de la proteína. El tiempo de almacenamiento no tuvo efecto sobre la población de hongos. El valor máximo de aflatoxina obtenido fue de 45,5 μg/kg y permaneció dentro de los limites aceptables para alimentación de bovinos. Conclusión: La deshidratación de WBG al sol es eficiente para garantizar la conservación del material y viabilizar su almacenamiento. El almacenamiento por 180 días de este subproducto seco no compromete su calidad nutricional y microbiológica.

Resumo Antecedentes: Os grãos de cervejaria são um subproduto que podem ser incluídos na dieta de ruminantes, no entanto sua forma úmida dificulta o armazenamento e a conservação desse produto. Objetivo: Avaliar a eficiência de desidratação ao sol dos grãos úmidos de cervejaria (WBG) e o efeito do período de armazenamento sobre a qualidade nutricional e microbiológica. Métodos: Para avaliar a desidratação do WBG, utilizou-se um delineamento experimental inteiramente casualizado, sendo os tratamentos os tempos de 0 e 1, 2, 4, 6, 8, 10, 12, 14 e 16 horas de exposição ao sol. Um segundo experimento foi realizado em delineamento inteiramente casualizado. Os tratamentos foram 0, 10, 20, 30, 60, 90, 120, 150 e 180 dias de armazenamento do subproduto seco. Resultados: A matéria seca (DM) do WBG apresentou efeito linear crescente com o processo de desidratação. A composição química dos grãos secos de cervejaria não apresentou efeito em função dos tempos de armazenagem. A proteína indigestível (Fração C) aumentou linearmente, no entanto não comprometeu a produção cumulativa de gás e a digestibilidade in vitro da DM e da proteína. A população de fungos não apresentou efeito com o tempo de armazenamento. O valor máximo de aflatoxina obtido foi de 45,5 μg/kg e permaneceu dentro dos limites aceitáveis para a alimentação de bovinos. Conclusão: A desidratação do WBG ao sol foi eficiente em garantir a conservação do material e viabilizar o seu armazenamento. A estocagem desse subproduto seco por 180 dias não comprometeu a sua qualidade nutricional e microbiológica.

Pax5 regulates B cell immunity by promoting PI3K signaling via PTEN down-regulation.

The transcription factor Pax5 controls B cell development, but its role in mature B cells is largely enigmatic. Here, we demonstrated that the loss of Pax5 by conditional mutagenesis in peripheral B lymphocytes led to the strong reduction of B-1a, marginal zone (MZ), and germinal center (GC) B cells as well as plasma cells. Follicular (FO) B cells tolerated the loss of Pax5 but had a shortened half-life. The Pax5-deficient FO B cells failed to proliferate upon B cell receptor or Toll-like receptor stimulation due to impaired PI3K-AKT signaling, which was caused by increased expression of PTEN, a negative regulator of the PI3K pathway. Pax5 restrained PTEN protein expression at the posttranscriptional level, likely involving Pten-targeting microRNAs. Additional PTEN loss in Pten,Pax5 double-mutant mice rescued FO B cell numbers and the development of MZ B cells but did not restore GC B cell formation. Hence, the posttranscriptional down-regulation of PTEN expression is an important function of Pax5 that facilitates the differentiation and survival of mature B cells, thereby promoting humoral immunity.

Mapping the Structure of Oxygen-Doped Wurtzite Aluminum Nitride Coatings from Ab Initio Random Structure Search and Experiments.

Machine learning is changing how we design and interpret experiments in materials science. In this work, we show how unsupervised learning, combined with ab initio random structure searching, improves our understanding of structural metastability in multicomponent alloys. We focus on the case of Al-O-N alloys where the formation of aluminum vacancies in wurtzite AlN upon the incorporation of substitutional oxygen can be seen as a general mechanism of solids where crystal symmetry is reduced to stabilize defects. The ideal AlN wurtzite crystal structure occupation cannot be matched due to the presence of an aliovalent hetero-element into the structure. The traditional interpretation of the c-lattice shrinkage in sputter-deposited Al-O-N films from X-ray diffraction (XRD) experiments suggests the existence of a solubility limit at 8 at % oxygen content. Here, we show that such naive interpretation is misleading. We support XRD data with accurate ab initio modeling and dimensionality reduction on advanced structural descriptors to map structure-property relationships. No signs of a possible solubility limit are found. Instead, the presence of a wide range of non-equilibrium oxygen-rich defective structures emerging at increasing oxygen contents suggests that the formation of grain boundaries is the most plausible mechanism responsible for the lattice shrinkage measured in Al-O-N sputtered films. We further confirm our hypothesis using positron annihilation lifetime spectroscopy.

Effects of Black Wattle (Acacia mearnsii) Condensed Tannins on Intake, Protozoa Population, Ruminal Fermentation, and Nutrient Digestibility in Jersey Steers.

The objective of this study was to evaluate the effect of inclusion of condensed tannins (CT) from black wattle (Acacia mearnsii) on feed intake, ruminal protozoa population, ruminal fermentation, and nutrient digestibility in Jersey steers. Five ruminally-cannulated steers were used in a 5x5 Latin square design, with five periods of 20 days each (14 days for diet adaptation and six days for sample collection per period). Treatments were composed of dietary inclusion levels of condensed tannins at 0, 5, 10, 15, and 20 g/kg of diet dry matter. Intakes of dry matter, organic matter, ether extract, crude protein, neutral detergent fiber, and total digestible nutrients were not affected by condensed tannins. The ruminal pH was reduced linearly with tannin levels. Ruminal ammonia nitrogen concentration was not affected by tannins. Tannins reduced the molar proportion of acetate and did not affect the ruminal protozoal population, which might be related to the low doses used. Digestibilities of dry matter, organic matter, and neutral detergent fiber were not altered; however, there was a linear reduction in crude protein digestibility. Based on these results, CT extracts from black wattle are not recommended for improving nutrient utilization in steers at the tested levels.

Efficacious Cefazolin Prophylactic Dose for Morbidly Obese Women Undergoing Bariatric Surgery Based on Evidence from Subcutaneous Microdialysis and Populational Pharmacokinetic Modeling.

PURPOSE: To determine the efficacious cefazolin prophylactic dose for bariatric surgery using free subcutaneous concentrations accessed by microdialysis after 2 g or 3 g i.v. bolus dosing to morbidly obese women and POPPK modeling. METHODS: A POPPK model with variable plasma and subcutaneous tissue protein binding was developed to simultaneously describe plasma and tissue data sets. The outcomes was predicted for common surgical site infection (SSI) bacteria over 3, 4, 5 and 6 h periods postdose, as probability of target attainment (PTA) using Monte Carlo simulation. RESULTS: CFZ 2 g warrant up to 5 h SSI prophylaxis for bacteria with MICs ≤1 mg/L such as Escherichia coli and Staphylococcus aureus. For species such as Klebsiella pneumoniae, which present MIC distribution frequency of 2 mg/L, the maintenance of PTA ≥ 90% occurs with a 3 g dose for surgeries lasting up to 5 h, and 2 g dose provide an adequate response up to 4 h (PTA of 89%). CONCLUSIONS: Effectiveness of CFZ 2 g is similar to 3 g against bacteria with a MIC up to 2 mg/L, especially if the surgery does not last for more than 4 h.

Control of B-1a cell development by instructive BCR signaling.

B-1a cells remain one of the most enigmatic lymphocyte subsets. In this review, we discuss recent advances in our understanding of the development of these cells and their regulation by the transcription factors Bhlhe41 and Arid3a as well as by the RNA-binding protein Lin28b. A large body of literature supports an instructive role of BCR signaling in B-1a cell development and lineage commitment, which is initiated only after signaling from an autoreactive BCR. While both fetal and adult hematopoiesis can generate B-1a cells, the contribution of adult hematopoiesis to the B-1a cell compartment is low under physiological conditions. We discuss several models that can reconcile the instructive role of BCR signaling with this fetal bias in B-1a cell development.

Orbital paraganglioma in a dog.

A 10-year-old Rottweiler presented with right-sided moderately painful exophthalmia, blindness, absence of dazzle and pupillary light reflexes, a swollen optic nerve head and ventrolateral indentation of the globe. On magnetic resonance imaging, a 3 x 2 x 2 cm mass with a fluid filled center and contrast-enhancing periphery was noted posteriolateral of the globe. Orbital ultrasound was used for a guided fine needle aspirate of the mass. Cytology revealed moderate numbers of polygonal cells with lightly basophilic cytoplasm. Several cells showed nuclear pseudoinclusions. Histopathology following exenteration of the orbit revealed an infiltrative, extradural neoplasm surrounding the optic nerve. Cells were arranged in packets. Neoplastic cells were immunopositive for neuron specific enolase, synaptophysin and chromogranin A and immunonegative for cytokeratin. Findings were consistent with an extra-adrenal paraganglioma (neuroendocrine tumour). Although complete excision could not be confirmed on histopathology, the owners reported no apparent tumour recurrence 25 months after surgery. In conclusion a paraganglioma should be considered as a differential diagnosis of an orbital mass.

Comparison of two combinations of opioid and non-opioid analgesics for acute periradicular abscess: a randomized clinical trial

Abstract Acute periradicular abscess is a condition characterized by the formation and propagation of pus in the periapical tissues and generally associated with debilitating pain. Objective: The aim of this study was to compare the overall analgesic effectiveness of two combinations of opioid and non-opioid analgesics for acute periradicular abscess. Material and Methods: This study included 26 patients who sought emergency care in a Brazilian dental school. The patients were randomly divided into two groups: Co/Ac - oral prescription of codeine (30 mg) plus acetaminophen (500 mg), every 4 h, for 3 days or Tr/Ac - oral prescription of tramadol hydrochloride (37.5 mg) plus acetaminophen (500 mg) on the same schedule. Two factors were evaluated: (1) pain scores recorded by the patients in a pain diary 6, 12, 24, 48, and 72 h after treatment, using the Visual Analogue Scale; and (2) the occurrence of adverse effects. Results: In both groups, there was a reduction in pain scores over time. For the Co/Ac group, there was a significant reduction in the scores 12, 24, 48, and 72 hours after treatment (P<0.05). In the Tr/Ac group, the scores significantly decreased over time from time point 6 h (P<0.05). Comparing the pain at each time point, the groups were not significantly different (P>0.05), i.e., both treatments were effective in controlling pain caused by APA; however, the combination of Tr/Ac caused more adverse reactions as two patients had to stop using the medication. Conclusion: This study suggests that, considering both analgesic efficacy and safety, the combination of codeine and acetaminophen is more effective to control moderate to severe pain from acute periradicular abscesses.

Molecular role of the PAX5-ETV6 oncoprotein in promoting B-cell acute lymphoblastic leukemia.

PAX5 is a tumor suppressor in B-ALL, while the role of PAX5 fusion proteins in B-ALL development is largely unknown. Here, we studied the function of PAX5-ETV6 and PAX5-FOXP1 in mice expressing these proteins from the Pax5 locus. Both proteins arrested B-lymphopoiesis at the pro-B to pre-B-cell transition and, contrary to their proposed dominant-negative role, did not interfere with the expression of most regulated Pax5 target genes. Pax5-Etv6, but not Pax5-Foxp1, cooperated with loss of the Cdkna2a/b tumor suppressors in promoting B-ALL development. Regulated Pax5-Etv6 target genes identified in these B-ALLs encode proteins implicated in pre-B-cell receptor (BCR) signaling and migration/adhesion, which could contribute to the proliferation, survival, and tissue infiltration of leukemic B cells. Together with similar observations made in human PAX5-ETV6+ B-ALLs, these data identified PAX5-ETV6 as a potent oncoprotein that drives B-cell leukemia development.

Integrative Analyses of Colorectal Cancer Show Immunoscore Is a Stronger Predictor of Patient Survival Than Microsatellite Instability.

Microsatellite instability in colorectal cancer predicts favorable outcomes. However, the mechanistic relationship between microsatellite instability, tumor-infiltrating immune cells, Immunoscore, and their impact on patient survival remains to be elucidated. We found significant differences in mutational patterns, chromosomal instability, and gene expression that correlated with patient microsatellite instability status. A prominent immune gene expression was observed in microsatellite-instable (MSI) tumors, as well as in a subgroup of microsatellite-stable (MSS) tumors. MSI tumors had increased frameshift mutations, showed genetic evidence of immunoediting, had higher densities of Th1, effector-memory T cells, in situ proliferating T cells, and inhibitory PD1-PDL1 cells, had high Immunoscores, and were infiltrated with mutation-specific cytotoxic T cells. Multivariate analysis revealed that Immunoscore was superior to microsatellite instability in predicting patients' disease-specific recurrence and survival. These findings indicate that assessment of the immune status via Immunoscore provides a potent indicator of tumor recurrence beyond microsatellite-instability staging that could be an important guide for immunotherapy strategies.

A novel methylation derivatization method for δ(18)O analysis of individual carbohydrates by gas chromatography/pyrolysis-isotope ratio mass spectrometry.

RATIONALE: The oxygen isotope ratio (δ(18)O) of carbohydrates derived from animals, plants, sediments, and soils provides important information about biochemical and physiological processes, past environmental conditions, and geographical origins, which are otherwise not available. Nowadays, δ(18)O analyses are often performed on carbohydrate bulk material, while compound-specific δ(18)O analyses remain challenging and methods for a wide range of individual carbohydrates are rare. METHODS: To improve the δ(18)O analysis of individual carbohydrates by gas chromatography/pyrolysis-isotope ratio mass spectrometry (GC/Pyr-IRMS) we developed a new methylation derivatization method. Carbohydrates were fully methylated within 24 h in an easy-to-handle one-pot reaction in acetonitrile, using silver oxide as proton acceptor, methyl iodide as methyl group carrier, and dimethyl sulfide as catalyst. RESULTS: The precision of the method ranged between 0.12 and 1.09‰ for the δ(18)O values of various individual carbohydrates of different classes (mono-, di-, and trisaccharides, alditols), with an accuracy of a similar order of magnitude, despite high variation in peak areas. Based on the δ(18)O values of the main isomers, important monosaccharides such as glucose and fructose could also be precisely analyzed for the first time. We tested the method on standard mixtures, honey samples, and leaf carbohydrates extracted from Pinus sylvestris, showing that the method is also applicable to different carbohydrate mixtures. CONCLUSIONS: The new methylation method shows unrivalled accuracy and precision for δ(18)O analysis of various individual carbohydrates; it is fast and easy-to-handle, and may therefore find wide-spread application.

Characterization of the immunophenotypes and antigenomes of colorectal cancers reveals distinct tumor escape mechanisms and novel targets for immunotherapy.

BACKGROUND: While large-scale cancer genomic projects are comprehensively characterizing the mutational spectrum of various cancers, so far little attention has been devoted to either define the antigenicity of these mutations or to characterize the immune responses they elicit. Here we present a strategy to characterize the immunophenotypes and the antigen-ome of human colorectal cancer. RESULTS: We apply our strategy to a large colorectal cancer cohort (n = 598) and show that subpopulations of tumor-infiltrating lymphocytes are associated with distinct molecular phenotypes. The characterization of the antigenome shows that a large number of cancer-germline antigens are expressed in all patients. In contrast, neo-antigens are rarely shared between patients, indicating that cancer vaccination requires individualized strategy. Analysis of the genetic basis of the tumors reveals distinct tumor escape mechanisms for the patient subgroups. Hypermutated tumors are depleted of immunosuppressive cells and show upregulation of immunoinhibitory molecules. Non-hypermutated tumors are enriched with immunosuppressive cells, and the expression of immunoinhibitors and MHC molecules is downregulated. Reconstruction of the interaction network of tumor-infiltrating lymphocytes and immunomodulatory molecules followed by a validation with 11 independent cohorts (n = 1,945) identifies BCMA as a novel druggable target. Finally, linear regression modeling identifies major determinants of tumor immunogenicity, which include well-characterized modulators as well as a novel candidate, CCR8, which is then tested in an orthologous immunodeficient mouse model. CONCLUSIONS: The immunophenotypes of the tumors and the cancer antigenome remain widely unexplored, and our findings represent a step toward the development of personalized cancer immunotherapies.

Antibiotic prophylaxis in obese patients submitted to bariatric surgery. A systematic review.

PURPOSE: To review the use of cefazolin in prophylaxis of surgical wound infection (SSI) in bariatric surgery (BS). METHODS: A systematic review was performed from October to November, 2013 using the following databases: The Cochrane Library, Medline, LILACS, and EMBASE. The inclusion criteria were randomized clinical trials and observational studies that were evaluated by two independent reviewers. RESULTS: Nine hundred and sixty one titles were recovered after preliminary analysis (title and abstract), seven studies remained for final analysis. There were three clinical trials (one with SSI, and two with antibiotic levels as the outcome), and four were observational studies (three cohorts and one case-control, all had SSI as the outcome). After administration of 1g or 2 g, levels of cefazolin in serum and tissue were suboptimal according to two studies. Results from observational studies indicated that different antibiotics were used for prophylaxis of SSI in BS and that use of other drugs may be associated with higher rates of SSI. CONCLUSION: The use of cefazolin for surgical wound infection prophylaxis in bariatric surgery is recommended, however further studies are needed in order to refine parameters as initial dose, redose, moment of administration and lasting of prophylaxis.

Antibiotic prophylaxis in obese patients submitted to bariatric surgery. A systematic review

To review the use of cefazolin in prophylaxis of surgical wound infection (SSI) in bariatric surgery (BS). METHODS: A systematic review was performed from October to November, 2013 using the following databases: The Cochrane Library, Medline, LILACS, and EMBASE. The inclusion criteria were randomized clinical trials and observational studies that were evaluated by two independent reviewers. RESULTS: Nine hundred and sixty one titles were recovered after preliminary analysis (title and abstract), seven studies remained for final analysis. There were three clinical trials (one with SSI, and two with antibiotic levels as the outcome), and four were observational studies (three cohorts and one case-control, all had SSI as the outcome). After administration of 1g or 2 g, levels of cefazolin in serum and tissue were suboptimal according to two studies. Results from observational studies indicated that different antibiotics were used for prophylaxis of SSI in BS and that use of other drugs may be associated with higher rates of SSI. CONCLUSION: The use of cefazolin for surgical wound infection prophylaxis in bariatric surgery is recommended, however further studies are needed in order to refine parameters as initial dose, redose, moment of administration and lasting of prophylaxis.

A survey of tools for variant analysis of next-generation genome sequencing data.

Recent advances in genome sequencing technologies provide unprecedented opportunities to characterize individual genomic landscapes and identify mutations relevant for diagnosis and therapy. Specifically, whole-exome sequencing using next-generation sequencing (NGS) technologies is gaining popularity in the human genetics community due to the moderate costs, manageable data amounts and straightforward interpretation of analysis results. While whole-exome and, in the near future, whole-genome sequencing are becoming commodities, data analysis still poses significant challenges and led to the development of a plethora of tools supporting specific parts of the analysis workflow or providing a complete solution. Here, we surveyed 205 tools for whole-genome/whole-exome sequencing data analysis supporting five distinct analytical steps: quality assessment, alignment, variant identification, variant annotation and visualization. We report an overview of the functionality, features and specific requirements of the individual tools. We then selected 32 programs for variant identification, variant annotation and visualization, which were subjected to hands-on evaluation using four data sets: one set of exome data from two patients with a rare disease for testing identification of germline mutations, two cancer data sets for testing variant callers for somatic mutations, copy number variations and structural variations, and one semi-synthetic data set for testing identification of copy number variations. Our comprehensive survey and evaluation of NGS tools provides a valuable guideline for human geneticists working on Mendelian disorders, complex diseases and cancers.

Co-expressed genes prepositioned in spatial neighborhoods stochastically associate with SC35 speckles and RNA polymerase II factories.

Chromosomally separated, co-expressed genes can be in spatial proximity, but there is still debate about how this nuclear organization is achieved. Proposed mechanisms include global genome organization, preferential positioning of chromosome territories, or gene-gene sharing of various nuclear bodies. To investigate this question, we selected a set of genes that were co-expressed upon differentiation of human multipotent stem cells. We applied a novel multi-dimensional analysis procedure which revealed that prior to gene expression, the relative position of these genes was conserved in nuclei. Upon stem cell differentiation and concomitant gene expression, we found that co-expressed genes were closer together. In addition, we found that genes in the same 1-µm-diameter neighborhood associated with either the same splicing speckle or to a lesser extent with the same transcription factory. Dispersal of speckles by overexpression of the serine-arginine (SR) protein kinase cdc2-like kinase Clk2 led to a significant drop in the number of genes in shared neighborhoods. We demonstrate quantitatively that the frequencies of speckle and factory sharing can be explained by assuming stochastic selection of a nuclear body within a restricted sub-volume defined by the original global gene positioning present prior to gene expression. We conclude that the spatial organization of these genes is a two-step process in which transcription-induced association with nuclear bodies enhances and refines a pre-existing global organization.

Effect of parathyroid hormone-related protein in an in vitro hypertrophy model for mesenchymal stem cell chondrogenesis.

PURPOSE: Mesenchymal stem cells (MSCs) express markers of hypertrophic chondrocytes during chondrogenic differentiation. We tested the suitability of parathyroid hormone-related protein (PTHrP), a regulator of chondrocyte hypertrophy in embryonic cartilage development, for the suppression of hypertrophy in an in vitro hypertrophy model of chondrifying MSCs. METHODS: Chondrogenesis was induced in human MSCs in pellet culture for two weeks and for an additional two weeks cultures were either maintained in standard chondrogenic medium or transferred to a hypertrophy-enhancing medium. PTHrP(1-40) was added to the medium throughout the culture period at concentrations from 1 to 1,000 pM. Pellets were harvested on days one, 14 and 28 for biochemical and histological analysis. RESULTS: Hypertrophic medium clearly enhanced the hypertrophic phenotype, with increased cell size, and strong alkaline phosphatase (ALP) and type X collagen staining. In chondrogenic medium, 1-100 pM PTHrP(1-40) did not inhibit chondrogenic differentiation, whereas 1,000 pM PTHrP(1-40) significantly reduced chondrogenesis. ALP activity was dose-dependently reduced by PTHrP(1-40) at 10-1,000 pM in chondrogenic conditions. Under hypertrophy-enhancing conditions, PTHrP(1-40) did not inhibit the induction of the hypertrophy. At the highest concentration (1,000 pM) in the hypertrophic group, aggregates were partially dedifferentiated and differentiated areas of these aggregates maintained their hypertrophic appearance. CONCLUSIONS: PTHrP(1-40) treatment dose-dependently reduced ALP expression in MSC pellets cultured under standard chondrogenic conditions and is thus beneficial for the maintenance of the chondrogenic phenotype in this medium condition. When cultured under hypertrophy-enhancing conditions, PTHrP(1-40) could not diminish the induced enhancement of hypertrophy in the MSC pellets.