Antiradical capacity assay for hydrophobic substances using hemin-catalyzed luminol chemiluminescence in cationic micelles.

Antioxidant substances which can diminish the steady-state concentration of free radicals in vivo are important in the human dietary to diminish the deleterious effects of oxidative stress. As the potential of certain substances as antioxidants is difficult to be verified in vivo, simple chemical in vitro assays which test the potential of substances as antioxidants are of great importance for the screening of new antioxidants. These assays measure the capacity of a substance to suppress free radicals. We describe here an antiradical capacity assay, based on luminol chemiluminescence, in cationic micellar medium, allowing the capacity determination of hydrophobic compounds. The antiradical capacity of antioxidants is determined using the Trolox standard by the measurement of the light emission inhibition area caused by the addition of different antiradical concentrations. The obtained results are compared to the values determined using the scavenging of stable free radicals be the substances and shown to be similar for compounds like uric acid, rutin, and quercetin. However, for vitamin E, the luminol assay results in a considerably higher antiradical capacity than the assay with a stable free radical, which is rationalized by the higher reactivity of the radical generated in the luminol assay and a specific localization of vitamin E in the micellar medium.

Ceftolozane/tazobactam for the treatment of Pseudomonas aeruginosa infections: A multicenter case series analysis / Ceftolozano/tazobactam para el tratamiento de infecciones por Pseudomonas aeruginosa: una serie de casos multicéntrica

Introduction Pseudomonas aeruginosa displays resistance to several available antibiotics. Infections caused by this pathogen are associated with a high mortality, morbidity, and considerable healthcare resource utilization and costs. This study was aimed at describing the use of ceftolozane/tazobactam (C/T) for the treatment of patients with P. aeruginosa infections. Methods Case series analysis of hospitalized patients treated with C/T for P. aeruginosa infections in five public Portuguese hospitals. Patients presenting with infections caused by this pathogen and receiving C/T for at least 72h during hospitalization were eligible. Results Sixty-four hospitalized patients with P. aeruginosa infections treated with C/T were evaluated between December 2016 and July 2019. Most patients were aged between 60 and 79 years (53.9%). Patients presented a total of 68 P. aeruginosa infections, with respiratory infections being the most common (28.1%, 18 out of 64). Most P. aeruginosa strains (85.9%, 55 out of 64) were extensively drug-resistant (XDR). C/T was mostly used as targeted therapy (98.4%, 63 out of 64 patients) and as monotherapy (72.7%, 47 out of 64 patients). Combination therapy was used in 47.4% (9 out of 19) of patients with bacteriemia. Most patients had successful microbiological (79.2%, 42 out of 53) and clinical (78.7%, 48 out of 61) outcomes. All-cause in-hospital mortality rate was 34.4%. Conclusion The present case series contributes to the body of evidence suggesting that C/T is an effective and safe option for treating P. aeruginosa infections, namely those caused by XDR strains, both when used as mono- or combination therapy (AU)

Introducción Pseudomonas aeruginosa presenta mecanismos de resistencia a diversos antibióticos. Las infecciones causadas por este patógeno aumentan la morbimortalidad, el uso de recursos y los costos asociados. Este estudio tuvo como objetivo describir el uso de ceftolozano/tazobactam (C/T) para el tratamiento de pacientes con infecciones por P. aeruginosa. Métodos Serie de casos de pacientes hospitalizados con infecciones por P. aeruginosa que fueron tratados con C/T en 5 hospitales portugueses. Fueron incluidos pacientes que presentaban infecciones por este patógeno y recibían terapéutica con C/T durante al menos 72 horas. Resultados Entre diciembre de 2016 y julio de 2019 fueron analizados 64 pacientes hospitalizados con infecciones por P. aeruginosa tratados con C/T. La mayoría tenían entre 60 y 79 años (53,9%). Se aislaron 68 infecciones por P. aeruginosa, siendo más frecuentes las respiratorias (28,1%; 18/64). La mayoría de las cepas de P. aeruginosa (85,9%; 55764) eran extremadamente resistentes. El C/T se utilizó principalmente como terapia dirigida (98,4%; 63/64 pacientes) y en monoterapia (72,7%; 47/64 pacientes). La terapia combinada se utilizó en el 47,4% (9/19) de los pacientes con bacteriemia. La mayoría de los pacientes tuvieron resultados microbiológicos (79,2%; 42/53) y clínicos (78,7%; 48/61) satisfactorios. La tasa de mortalidad intrahospitalaria por todas las causas fue del 34,4%. Conclusión La presente serie de casos sustenta que la terapéutica con C/T es una alternativa efectiva y segura para las infecciones por P. aeruginosa, particularmente por cepas extremadamente resistentes, en monoterapia o en terapia combinada (AU)

Ceftolozane/tazobactam for the treatment of Pseudomonas aeruginosa infections: A multicenter case series analysis.

INTRODUCTION: Pseudomonas aeruginosa displays resistance to several available antibiotics. Infections caused by this pathogen are associated with a high mortality, morbidity, and considerable healthcare resource utilization and costs. This study was aimed at describing the use of ceftolozane/tazobactam (C/T) for the treatment of patients with P. aeruginosa infections. METHODS: Case series analysis of hospitalized patients treated with C/T for P. aeruginosa infections in five public Portuguese hospitals. Patients presenting with infections caused by this pathogen and receiving C/T for at least 72h during hospitalization were eligible. RESULTS: Sixty-four hospitalized patients with P. aeruginosa infections treated with C/T were evaluated between December 2016 and July 2019. Most patients were aged between 60 and 79 years (53.9%). Patients presented a total of 68 P. aeruginosa infections, with respiratory infections being the most common (28.1%, 18 out of 64). Most P. aeruginosa strains (85.9%, 55 out of 64) were extensively drug-resistant (XDR). C/T was mostly used as targeted therapy (98.4%, 63 out of 64 patients) and as monotherapy (72.7%, 47 out of 64 patients). Combination therapy was used in 47.4% (9 out of 19) of patients with bacteriemia. Most patients had successful microbiological (79.2%, 42 out of 53) and clinical (78.7%, 48 out of 61) outcomes. All-cause in-hospital mortality rate was 34.4%. CONCLUSION: The present case series contributes to the body of evidence suggesting that C/T is an effective and safe option for treating P. aeruginosa infections, namely those caused by XDR strains, both when used as mono- or combination therapy.

Integration analyses of structural variations and differential gene expression associated with beef fatty acid profile in Nellore cattle.

This study aimed to integrate analyses of structural variations and differentially expressed genes (DEGs) associated with the beef fatty acid (FA) profile in Nellore cattle. Copy numbers variation (CNV) detection was performed using the penncnv algorithm and CNVRuler software in 3794 genotyped animals through the High-Density Bovine BeadChip. In order to perform the genomic wide association study (GWAS), a total of 963 genotyped animals were selected to obtain the intramuscular lipid concentration and quantify the beef FA profile. A total of 48 animals belonging to the same farm and management lot were extracted from the 963 genotyped and phenotyped animals to carry out the transcriptomic and differentially expressed gene analyses. The GWAS with extreme groups of FA profiles was performed using a logistic model. A total of 43, 42, 66 and 35 significant CNV regions (p < 0.05) for saturated, monounsaturated, polyunsaturated and omega 3 and 6 fatty acids were identified respectively. The paired-end sequencing of 48 samples was performed using the Illumina HiSeq2500 platform. Real-time quantitative PCR was used to validate the DEGs identified by RNA-seq analysis. The results showed several DEGs associated with the FA profile of Longissimus thoracis, such as BSCL2 and SAMD8. Enriched terms as the cellular response to corticosteroid (GO:0071384) and glucocorticoid stimulus (GO:0071385) could be highlighted. The identification of structural variations harboring candidate genes for beef FA must contribute to the elucidation of the genetic basis that determines the beef FA composition of intramuscular fat in Nellore cattle. Our results will contribute to the identification of potential biomarkers for complex phenotypes, such as the FA profile, to improve the reliability of the genomic predictions including pre-selected variants using differentiated weighting in the genomic models.

Screening a Targeted Panel of Genes by Next-Generation Sequencing Improves Risk Stratification in Real World Patients with Acute Myeloid Leukemia.

Although mutation profiling of defined genes is recommended for classification of acute myeloid leukemia (AML) patients, screening of targeted gene panels using next-generation sequencing (NGS) is not always routinely used as standard of care. The objective of this study was to prospectively assess whether extended molecular monitoring using NGS adds clinical value for risk assessment in real-world AML patients. We analyzed a cohort of 268 newly diagnosed AML patients. We compared the prognostic stratification of our study population according to the European LeukemiaNet recommendations, before and after the incorporation of the extended mutational profile information obtained by NGS. Without access to NGS data, 63 patients (23%) failed to be stratified into risk groups. After NGS data, only 27 patients (10%) failed risk stratification. Another 33 patients were re-classified as adverse-risk patients once the NGS data was incorporated. In total, access to NGS data refined risk assessment for 62 patients (23%). We further compared clinical outcomes with prognostic stratification, and observed unexpected outcomes associated with FLT3 mutations. In conclusion, this study demonstrates the prognostic utility of screening AML patients for multiple gene mutations by NGS and underscores the need for further studies to refine the current risk classification criteria.

Evaluation of 2 Low-cost High-definition Video Systems for Venous Anastomosis.

Background. A literature review of the use of video systems for magnification has suggested that so far, this novel magnification system has only been used to perform arterial anastomoses. The aim of this study was to evaluate the use of 2 low-cost methods of video-assisted magnification in microvascular venous anastomosis in rats. Methods. Thirty rats were randomly divided into 3 matched groups according to the magnification system used: the microscope group, with venous anastomosis performed under a microscope; the camcorder system group, with the procedures performed under a high-definition Handycam HDR-XR160; and the Photographic camera group, for which procedures were performed with an EOS Rebel T3i photographic camera. In both video system groups, a magnification system was connected to a 42-in. television by an HDMI cable. We analyzed weight, venous caliber, total surgery and anastomosis time, patency immediately and 14 days postoperatively, number of stitches, and histological analyses. Results. There were no significant differences between the groups in weight, venous caliber, or number of stitches. Anastomosis under the video systems took longer. Patency rates were similar between the groups, except for the photographic system group that has a lower patency rate at 14 days. The histological analyses were similar in all groups. Conclusion. It is possible to perform a venous anastomosis in rats through video system magnification, with a satisfactory success rate comparable with that for procedures performed under microscopes; however, the kind of video system has a great influence on the final patency.

Genome-wide scan for runs of homozygosity in the composite Montana Tropical® beef cattle.

The aim of this study was to assess the distribution of runs of homozygosity (ROH) and autozygosity islands in the composite Montana Tropical® beef cattle to explore hotspot regions which could better characterize the different biological types within the composite breed. Montana animals (n = 1,436) were genotyped with the GGP-LD BeadChip (~30,000 markers). ROH was identified in every individual using the plink v1.90 software. Medium and long ROH prevailed in the genome, which accounted for approximately 74% of all ROH detected. On an average, 2.0% of the genome was within ROH, agreeing with the pedigree-based inbreeding coefficient. The Montana cattle with a higher proportion of productive breed types showed the highest number of autozygosity islands (n = 17), followed by those with a higher proportion of breeds adapted to tropical environments (n = 15). Enriched terms (p < .05) associated with the immune and inflammatory response, homeostasis, reproduction, mineral absorption, and lipid metabolism were described within the autozygosity islands. In this regard, over-represented GO terms and KEGG pathways described in this population may play a key role in providing information to explore the genetic and biological mechanisms together with the genomic regions underlying each biological type that favoured their optimal performance ability in tropical and subtropical regions.

Genetic association between different criteria to define sexual precocious heifers with growth, carcass, reproductive and feed efficiency indicator traits in Nellore cattle using genomic information.

The aim of this study was to estimate genetic parameters for different precocious calving criteria and their relationship with reproductive, growth, carcass and feed efficiency in Nellore cattle using the single-step genomic BLUP. The reproductive traits used were probability of precocious calving (PPC) at 24 (PPC24), 26 (PPC26), 28 (PPC28) and 30 (PPC30) months of age, stayability (STAY) and scrotal circumference at 455 days of age (SC455). Growth traits such as weights at 240 (W240) and 455 (W455) days of age and adult weight (AW) were used. Rib eye area (REA), subcutaneous fat thickness (SFT), rump fat thickness (RFT) and residual feed intake (RFI) were included in the analyses. The estimation of genetic parameters was performed using a bi-trait threshold model including genomic information in a single-step approach. Heritability for PPC traits was moderate to high (0.29-0.56) with highest estimates for PPC24 (0.56) and PPC26 (0.50). Genetic correlation estimates between PPC and STAY weakened as a function of calving age. Correlation with SC455, growth and carcass traits were low (0.25-0.31; -0.22 to 0.04; -0.09 to 0.18, respectively), the same occurs with RFI (-0.09 to 0.08), this suggests independence between female sexual precocity and feed efficiency traits. The results of this study encourage the use of PPC traits in Nellore cattle because the selection for such trait would not have a negative impact on reproductive, growth, carcass and feed efficiency indicator traits. Stayability for sexual precocious heifers (PPC24 and PPC26) must be redefined to avoid incorrectly phenotype assignment.

A novel BCR-ABL1 mutation in a patient with Philadelphia chromosome-positive B-cell acute lymphoblastic leukemia.

Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL) represents the most common genetic subtype of adult ALL (20%-30%) and accounts for approximately 50% of all cases in the elderly. It has been considered the subgroup of ALL with the worst outcome. The introduction of tyrosine kinase inhibitors (TKIs) allows complete hematologic remission virtually in all patients, with improved disease-free survival and overall survival. Nevertheless, the emergence of resistant mutations in BCR-ABL1 may require different TKI strategies to overcome the patient's resistance and disease relapse. Here, we report a Ph+B-ALL case with persistent minimal residual disease (MRD) after treatment with dasatinib. The patient expressed the P190BCR-ABL1 isoform and a novel BCR-ABL1 mutation, p.Y440C. The latter is in the C-terminal lobe of the kinase domain, which likely induces deviations in the protein structure and activity and destabilizes its inactive conformation. The treatment was substituted by bosutinib, which binds to the active conformation of the protein, prior to allogeneic bone marrow transplant to overcome the lack of a complete response to dasatinib. These findings strengthen the importance of BCR-ABL1 mutational screening in Ph+ patients, particularly for those who do not achieve complete molecular remission.

Autozygosity islands and ROH patterns in Nellore lineages: evidence of selection for functionally important traits.

BACKGROUND: The aim of this study was to assess genome-wide autozygosity in a Nellore cattle population and to characterize ROH patterns and autozygosity islands that may have occurred due to selection within its lineages. It attempts also to compare estimates of inbreeding calculated from ROH (FROH), genomic relationship matrix (FGRM), and pedigree-based coefficient (FPED). RESULTS: The average number of ROH per animal was 55.15 ± 13.01 with an average size of 3.24 Mb. The Nellore genome is composed mostly by a high number of shorter segments accounting for 78% of all ROH, although the proportion of the genome covered by them was relatively small. The genome autozygosity proportion indicates moderate to high inbreeding levels for classical standards, with an average value of 7.15% (178.70 Mb). The average of FPED and FROH, and their correlations (- 0.05 to 0.26) were low. Estimates of correlation between FGRM-FPED was zero, while the correlation (- 0.01 to - 0.07) between FGRM-FROH decreased as a function of ROH length, except for FROH > 8Mb (- 0.03). Overall, inbreeding coefficients were not high for the genotyped animals. Autozygosity islands were evident across the genome (n = 62) and their genomic location did not largely differ within lineages. Enriched terms (p < 0.01) associated with defense response to bacteria (GO:0042742), immune complex reaction (GO:0045647), pregnancy-associated glycoproteins genes (GO:0030163), and organism growth (GO:0040014) were described within the autozygotic islands. CONCLUSIONS: Low FPED-FROH correlation estimates indicate that FPED is not the most suitable method for capturing ancient inbreeding when the pedigree does not extend back many generations and FROH should be used instead. Enriched terms (p < 0.01) suggest a strong selection for immune response. Non-overlapping islands within the lineages greatly explain the mechanism underlying selection for functionally important traits in Nellore cattle.

Genomic prediction ability for beef fatty acid profile in Nelore cattle using different pseudo-phenotypes.

The aim of the present study was to compare the predictive ability of SNP-BLUP model using different pseudo-phenotypes such as phenotype adjusted for fixed effects, estimated breeding value, and genomic estimated breeding value, using simulated and real data for beef FA profile of Nelore cattle finished in feedlot. A pedigree with phenotypes and genotypes of 10,000 animals were simulated, considering 50% of multiple sires in the pedigree. Regarding to phenotypes, two traits were simulated, one with high heritability (0.58), another with low heritability (0.13). Ten replicates were performed for each trait and results were averaged among replicates. A historical population was created from generation zero to 2020, with a constant size of 2000 animals (from generation zero to 1000) to produce different levels of linkage disequilibrium (LD). Therefore, there was a gradual reduction in the number of animals (from 2000 to 600), producing a "bottleneck effect" and consequently, genetic drift and LD starting in the generation 1001 to 2020. A total of 335,000 markers (with MAF greater or equal to 0.02) and 1000 QTL were randomly selected from the last generation of the historical population to generate genotypic data for the test population. The phenotypes were computed as the sum of the QTL effects and an error term sampled from a normal distribution with zero mean and variance equal to 0.88. For simulated data, 4000 animals of the generations 7, 8, and 9 (with genotype and phenotype) were used as training population, and 1000 animals of the last generation (10) were used as validation population. A total of 937 Nelore bulls with phenotype for fatty acid profiles (Sum of saturated, monounsaturated, omega 3, omega 6, ratio of polyunsaturated and saturated and polyunsaturated fatty acid profile) were genotyped using the Illumina BovineHD BeadChip (Illumina, San Diego, CA) with 777,962 SNP. To compare the accuracy and bias of direct genomic value (DGV) for different pseudo-phenotypes, the correlation between true breeding value (TBV) or DGV with pseudo-phenotypes and linear regression coefficient of the pseudo-phenotypes on TBV for simulated data or DGV for real data, respectively. For simulated data, the correlations between DGV and TBV for high heritability traits were higher than obtained with low heritability traits. For simulated and real data, the prediction ability was higher for GEBV than for Yc and EBV. For simulated data, the regression coefficient estimates (b(Yc,DGV)), were on average lower than 1 for high and low heritability traits, being inflated. The results were more biased for Yc and EBV than for GEBV. For real data, the GEBV displayed less biased results compared to Yc and EBV for SFA, MUFA, n-3, n-6, and PUFA/SFA. Despite the less biased results for PUFA using the EBV as pseudo-phenotype, the b(Yi,DGV estimates obtained for the different pseudo-phenotypes (Yc, EBV and GEBV) were very close. Genomic information can assist in improving beef fatty acid profile in Zebu cattle, since the use of genomic information yielded genomic values for fatty acid profile with accuracies ranging from low to moderate. Considering both simulated and real data, the ssGBLUP model is an appropriate alternative to obtain more reliable and less biased GEBVs as pseudo-phenotype in situations of missing pedigree, due to high proportion of multiple sires, being more adequate than EBV and Yc to predict direct genomic value for beef fatty acid profile.

Association study between copy number variation and beef fatty acid profile of Nellore cattle.

The aim of this study was to analyze the association between the copy number variation regions (CNVRs) and fatty acid profile phenotypes for saturated (SFA), monosaturated (MUFA), polyunsaturated (PUFA), ω6 and ω3 fatty acids, PUFA/SFA and ω6/ω3 ratios, as well as for their sums, in Nellore cattle (Bos primigenius indicus). A total of 963 males were finished in feedlot and slaughtered with approximately 2 years of age. Animals were genotyped with the BovineHD BeadChip (Illumina Inc., San Diego, CA, USA). The copy number variation (CNV) detection was performed using the PennCNV algorithm. Log R ratio (LRR) and allele B frequency (BAF) were used to estimate the CNVs. The association analyses were done using the CNVRuler software and applying a logistic regression model. The phenotype was adjusted using a linear model considering the fixed effects of contemporary group and the animal age at slaughter. The fatty acid profile was analyzed on samples of longissimus thoracis muscle using gas chromatography with a 100-m capillary column. For the association analysis, the adjusted phenotypic values were considered for the traits, while the data was adjusted for the effects of the farm and year of birth, management groups at birth, weaning, and superannuation. A total of 186 CNVRs were significant for SFA (43), MUFA (42), PUFA (66), and omega fatty acid (35) groups, totaling 278 known genes. On the basis of the results, several genes were associated with several fatty acids of different saturations. Olfactory receptor genes were associated with C12:0, C14:0, and C18:0 fatty acids. The SAMD8 and BSCL2 genes, both related to lipid metabolic process, were associated with C12:0. The RAPGEF6 gene was found to be associated with C18:2 cis-9 cis-12 n-6, and its function is related to regulation of GTPase activity. Among the results, we highlighted the olfactory receptor activity (GO:0004984), G-protein-coupled receptor activity (GO:0004930), potassium:proton antiporter activity (GO:0015386), sodium:proton antiporter activity (GO:0015385), and odorant-binding (GO:0005549) molecular functions. A large number of genes associated with fatty acid profile within the CNVRs were identified in this study. These findings must contribute to better elucidate the genetic mechanism underlying the fatty acid profile of intramuscular fat in Nellore cattle.

Intracellular Delivery of HCV NS3p gene using vectored particles.

Viral hepatitis caused by the hepatitis C virus (HCV) affects millions of people worldwide. The non-structural protein 3 (NS3), one of the most conserved proteins in HCV, is the target of many therapeutic studies. The NS3 protease domain (NS3p) has a range of cytotoxic T lymphocyte (CTL) epitopes, and synthesizing the protein inside the cells is the most appropriate way to present it to the immune system. We developed a tool to study this kind of presentation, using two vectored particle (VP) systems, one based on the Semliki Forest virus (SFV) and the other on HCV pseudoparticles (HCVpp), both carrying the protease domain of the NS3 gene. In addition to producing the particles, we developed a method to quantify these VPs using qRT-PCR. We produced batches of approximately 2.4 × 104 SFV-NS3p/µL and 4.0 × 102 HCVpp-NS3p/µL. BHK-21 and HuH-7 cells treated with the VPs expressed the NS3 protein, thus showing the functionality of this system.

Axillary nerve neurotization by a triceps motor branch: comparison between axillary and posterior arm approaches / Neurotização do nervo axilar por um ramo do tríceps: comparação entre acesso axilar e posterior

ABSTRACT Objectives: This study is aimed at comparing the functional outcome of axillary nerve neurotization by a triceps motor branch through the axillary approach and posterior arm approach. Methods: The study included 27 patients with post-traumatic brachial plexus injury treated with axillary nerve neurotization by a triceps motor branch for functional recovery of shoulder abduction and external rotation. The patients were retrospectively evaluated and two groups were identified, one with 13 patients undergoing axillary nerve neurotization by an axillary approach and the second with 14 patients using the posterior arm approach. Patients underwent assessment of muscle strength using the scale recommended by the British Medical Research Council, preoperatively and 18 months postoperatively, with useful function recovery considered as grade M3 or greater. Results: In the axillary approach group, 76.9% of patients achieved useful abduction function recovery and 69.2% achieved useful external rotation function recovery. In the group with posterior arm approach, 71.4% of patients achieved useful abduction function recovery and 50% achieved useful external rotation function recovery. The difference between the two groups was not statistically significant (p = 1.000 for the British Medical Research Council abduction scale and p = 0.440 for external rotation). Conclusion: According to the British Medical Research Council grading, axillary nerve neurotization with a triceps motor branch using axillary approach or posterior arm approach shows no statistical differences.

RESUMO Objetivos: Comparar o resultado funcional da neurotização do nervo axilar por um ramo motor do tríceps através do acesso axilar e do acesso posterior. Métodos: Foram incluídos no estudo 27 pacientes com lesão pós-traumática de plexo braquial submetidos à neurotização do nervo axilar por um ramo motor do tríceps para recuperação funcional do ombro de 2010 a 2014. Os pacientes foram avaliados e dois grupos foram identificados, um com 13 pacientes submetidos a neurotização do nervo axilar por um acesso axilar e o segundo com 14 pacientes nos quais foi usada a via de acesso posterior. Os pacientes foram submetidos a avaliação da força muscular com a escala preconizada pelo British Medical Research Council (BMRC) no pré-operatório e com 18 meses de pós-operatório, foi considerada força motora efetiva graduação M3 ou maior. Resultados: No grupo que fez o acesso axilar, 76,9% dos pacientes obtiveram força motora efetiva de abdução e 69,2% de rotação externa. Já no grupo com acesso posterior, 71,4% dos pacientes conseguiram força motora efetiva de abdução e 50% de rotação externa. A diferença entre os dois grupos não foi estatisticamente significante (p = 1,000 para escala BMRC de abdução e p = 0,440 para rotação externa). Conclusão: Na avaliação da graduação de força na escala BRMC, o uso do acesso axilar para neurotização de um ramo motor do tríceps para o nervo axilar não apresenta diferenças estatísticas em relação ao uso do acesso posterior.

Axillary nerve neurotization by a triceps motor branch: comparison between axillary and posterior arm approaches.

OBJECTIVES: This study is aimed at comparing the functional outcome of axillary nerve neurotization by a triceps motor branch through the axillary approach and posterior arm approach. METHODS: The study included 27 patients with post-traumatic brachial plexus injury treated with axillary nerve neurotization by a triceps motor branch for functional recovery of shoulder abduction and external rotation. The patients were retrospectively evaluated and two groups were identified, one with 13 patients undergoing axillary nerve neurotization by an axillary approach and the second with 14 patients using the posterior arm approach. Patients underwent assessment of muscle strength using the scale recommended by the British Medical Research Council, preoperatively and 18 months postoperatively, with useful function recovery considered as grade M3 or greater. RESULTS: In the axillary approach group, 76.9% of patients achieved useful abduction function recovery and 69.2% achieved useful external rotation function recovery. In the group with posterior arm approach, 71.4% of patients achieved useful abduction function recovery and 50% achieved useful external rotation function recovery. The difference between the two groups was not statistically significant (p = 1.000 for the British Medical Research Council abduction scale and p = 0.440 for external rotation). CONCLUSION: According to the British Medical Research Council grading, axillary nerve neurotization with a triceps motor branch using axillary approach or posterior arm approach shows no statistical differences.

OBJETIVOS: Comparar o resultado funcional da neurotização do nervo axilar por um ramo motor do tríceps através do acesso axilar e do acesso posterior. MÉTODOS: Foram incluídos no estudo 27 pacientes com lesão pós-traumática de plexo braquial submetidos à neurotização do nervo axilar por um ramo motor do tríceps para recuperação funcional do ombro de 2010 a 2014. Os pacientes foram avaliados e dois grupos foram identificados, um com 13 pacientes submetidos a neurotização do nervo axilar por um acesso axilar e o segundo com 14 pacientes nos quais foi usada a via de acesso posterior. Os pacientes foram submetidos a avaliação da força muscular com a escala preconizada pelo British Medical Research Council no pré-operatório e com 18 meses de pós-operatório, foi considerada força motora efetiva graduação M3 ou maior. RESULTADOS: No grupo que fez o acesso axilar, 76,9% dos pacientes obtiveram força motora efetiva de abdução e 69,2% de rotação externa. Já no grupo com acesso posterior, 71,4% dos pacientes conseguiram força motora efetiva de abdução e 50% de rotação externa. A diferença entre os dois grupos não foi estatisticamente significante (p = 1,000 para escala British Medical Research Council de abdução e p = 0,440 para rotação externa). CONCLUSÃO: Na avaliação da graduação de força na escala British Medical Research Council, o uso do acesso axilar para neurotização de um ramo motor do tríceps para o nervo axilar não apresenta diferenças estatísticas em relação ao uso do acesso posterior.

Intracellular delivery of HCV NS3p gene using vectored particles

Viral hepatitis caused by the hepatitis C virus (HCV) affects millions of people worldwide. The non-structural protein 3 (NS3), one of the most conserved proteins in HCV, is the target of many therapeutic studies. The NS3 protease domain (NS3p) has a range of cytotoxic T lymphocyte (CTL) epitopes, and synthesizing the protein inside the cells is the most appropriate way to present it to the immune system. We developed a tool to study this kind of presentation, using two vectored particle (VP) systems, one based on the Semliki Forest virus (SFV) and the other on HCV pseudoparticles (HCVpp), both carrying the protease domain of the NS3 gene. In addition to producing the particles, we developed a method to quantify these VPs using qRT-PCR. We produced batches of approximately 2.4x10(4) SFV-NS3p/mu L and 4.0x10(2) HCVpp-NS3p/mu L. BHK-21 and HuH-7 cells treated with the VPs expressed the NS3 protein, thus showing the functionality of this system.

Intracellular Delivery of HCV NS3p gene using vectored particles

Viral hepatitis caused by the hepatitis C virus (HCV) affects millions of people worldwide. The non-structural protein 3 (NS3), one of the most conserved proteins in HCV, is the target of many therapeutic studies. The NS3 protease domain (NS3p) has a range of cytotoxic T lymphocyte (CTL) epitopes, and synthesizing the protein inside the cells is the most appropriate way to present it to the immune system. We developed a tool to study this kind of presentation, using two vectored particle (VP) systems, one based on the Semliki Forest virus (SFV) and the other on HCV pseudoparticles (HCVpp), both carrying the protease domain of the NS3 gene. In addition to producing the particles, we developed a method to quantify these VPs using qRT-PCR. We produced batches of approximately 2.4x10(4) SFV-NS3p/mu L and 4.0x10(2) HCVpp-NS3p/mu L. BHK-21 and HuH-7 cells treated with the VPs expressed the NS3 protein, thus showing the functionality of this system.

Application of single step genomic BLUP under different uncertain paternity scenarios using simulated data.

The objective of this study was to investigate the application of BLUP and single step genomic BLUP (ssGBLUP) models in different scenarios of paternity uncertainty with different strategies of scaling the G matrix to match the A22 matrix, using simulated data for beef cattle. Genotypes, pedigree, and phenotypes for age at first calving (AFC) and weight at 550 days (W550) were simulated using heritabilities based on real data (0.12 for AFC and 0.34 for W550). Paternity uncertainty scenarios using 0, 25, 50, 75, and 100% of multiple sires (MS) were studied. The simulated genome had a total length of 2,333 cM, containing 735,293 biallelic markers and 7,000 QTLs randomly distributed over the 29 BTA. It was assumed that QTLs explained 100% of the genetic variance. For QTL, the amount of alleles per loci randomly ranged from two to four. The BLUP model that considers phenotypic and pedigree data, and the ssGBLUP model that combines phenotypic, pedigree and genomic information were used for genetic evaluations. Four ways of scaling the mean of the genomic matrix (G) to match to the mean of the pedigree relationship matrix among genotyped animals (A22) were tested. Accuracy, bias, and inflation were investigated for five groups of animals: ALL = all animals; BULL = only bulls; GEN = genotyped animals; FEM = females; and YOUNG = young males. With the BLUP model, the accuracies of genetic evaluations decreased for both traits as the proportion of unknown sires in the population increased. The EBV accuracy reduction was higher for GEN and YOUNG groups. By analyzing the scenarios for YOUNG (from 0 to 100% of MS), the decrease was 87.8 and 86% for AFC and W550, respectively. When applying the ssGBLUP model, the accuracies of genetic evaluation also decreased as the MS in the pedigree for both traits increased. However, the accuracy reduction was less than those observed for BLUP model. Using the same comparison (scenario 0 to 100% of MS), the accuracies reductions were 38 and 44.6% for AFC and W550, respectively. There were no differences between the strategies for scaling the G matrix for ALL, BULL, and FEM groups under the different scenarios with missing pedigree. These results pointed out that the uninformative part of the A22 matrix and genotyped animals with paternity uncertainty did not influence the scaling of G matrix. On the basis of the results, it is important to have a G matrix in the same scale of the A22 matrix, especially for the evaluation of young animals in situations with missing pedigree information. In these situations, the ssGBLUP model is an appropriate alternative to obtain a more reliable and less biased estimate of breeding values, especially for young animals with few or no phenotypic records. For accurate and unbiased genomic predictions with ssGBLUP, it is necessary to assure that the G matrix is compatible with the A22 matrix, even in situations with paternity uncertainty.

High-Definition Video System for Peripheral Neurorrhaphy in Rats.

BACKGROUND: Although all microsurgeries are based on the use of surgical microscopes, several alternative magnification systems have shown promising results. Improvements in image quality facilitated the use of video systems in microsurgeries with safety and accuracy. The aim of this study was to evaluate the use of a low-cost, video-assisted magnification system in peripheral neurorrhaphy in rats. METHODS: Twenty Wistar rats were randomly divided into 2 matched groups according to the magnification system used: the microscope group, with neurorrhaphy performed under a microscope with an image magnification of 40×; and the video system group, with the procedures performed under a video system composed of a high-definition Sony camcorder DCR-SR42 set to 52× magnification, macro lenses, 42-inch television, and a digital HDMI cable. We analyzed weight, nerve caliber, total surgery time, neurorrhaphy time, number of stitches, and number of axons in both ends (proximal and distal). RESULTS: There were no significant differences between groups in weight, nerve caliber, or number of stitches. Neurorrhaphy under the video system took longer (video: 5.60 minutes; microscope: 3.20 minutes; P < .05). Number of axons was similar between groups, both in proximal and distal stumps. CONCLUSION: It is possible to perform a peripheral neurorrhaphy in rats through video system magnification, but with a longer surgical time.

Genomic prediction for beef fatty acid profile in Nellore cattle.

The objective of this study was to compare SNP-BLUP, BayesCπ, BayesC and Bayesian Lasso methodologies to predict the direct genomic value for saturated, monounsaturated, and polyunsaturated fatty acid profile, omega 3 and 6 in the Longissimus thoracis muscle of Nellore cattle finished in feedlot. A total of 963 Nellore bulls with phenotype for fatty acid profiles, were genotyped using the Illumina BovineHD BeadChip (Illumina, San Diego, CA) with 777,962 SNP. The predictive ability was evaluated using cross validation. To compare the methodologies, the correlation between DGV and pseudo-phenotypes was calculated. The accuracy varied from -0.40 to 0.62. Our results indicate that none of the methods excelled in terms of accuracy, however, the SNP-BLUP method allows obtaining less biased genomic evaluations, thereby; this method is more feasible when taking into account the analyses' operating cost. Despite the lowest bias observed for EBV, the adjusted phenotype is the preferred pseudophenotype considering the genomic prediction accuracies regarding the context of the present study.