EEF1A1 transcription cofactor gene polymorphism is associated with muscle gene expression and residual feed intake in Nelore cattle.

Cis-acting effects of noncoding variants on gene expression and regulatory molecules constitute a significant factor for phenotypic variation in complex traits. To provide new insights into the impacts of single-nucleotide polymorphisms (SNPs) on transcription factors (TFs) and transcription cofactors (TcoF) coding genes, we carried out a multi-omic analysis to identify cis-regulatory effects of SNPs on these genes' expression in muscle and describe their association with feed efficiency-related traits in Nelore cattle. As a result, we identified one SNP, the rs137256008C > T, predicted to impact the EEF1A1 gene expression (ß = 3.02; P-value = 3.51E-03) and the residual feed intake trait (ß = - 3.47; P-value = 0.02). This SNP was predicted to modify transcription factor sites and overlaps with several QTL for feed efficiency traits. In addition, co-expression network analyses showed that animals containing the T allele of the rs137256008 SNP may be triggering changes in the gene network. Therefore, our analyses reinforce and contribute to a better understanding of the biological mechanisms underlying gene expression control of feed efficiency traits in bovines. The cis-regulatory SNP can be used as biomarker for feed efficiency in Nelore cattle.

A single nucleotide polymorphism in NEUROD1 is associated with production traits in Nelore beef cattle.

Feed efficiency and carcass characteristics are late-measured traits. The detection of molecular markers associated with them can help breeding programs to select animals early in life, and to predict breeding values with high accuracy. The objective of this study was to identify polymorphisms in the functional and positional candidate gene NEUROD1 (neurogenic differentiation 1), and investigate their associations with production traits in reference families of Nelore cattle. A total of 585 steers were used, from 34 sires chosen to represent the variability of this breed. By sequencing 14 animals with extreme residual feed intake (RFI) values, seven single nucleotide polymorphisms (SNPs) in NEUROD1 were identified. The investigation of marker effects on the target traits RFI, backfat thickness (BFT), ribeye area (REA), average body weight (ABW), and metabolic body weight (MBW) was performed with a mixed model using the restricted maximum likelihood method. SNP1062, which changes cytosine for guanine, had no significant association with RFI or REA. However, we found an additive effect on ABW (P ≤ 0.05) and MBW (P ≤ 0.05), with an estimated allele substitution effect of -1.59 and -0.93 kg0.75, respectively. A dominant effect of this SNP for BFT was also found (P ≤ 0.010). Our results are the first that identify NEUROD1 as a candidate that affects BFT, ABW, and MBW. Once confirmed, the inclusion of this SNP in dense panels may improve the accuracy of genomic selection for these traits in Nelore beef cattle as this SNP is not currently represented on SNP chips.

Allele- and parent-of-origin-specific effects on expression of the KCNJ11 gene: A candidate for meat tenderness in cattle.

In contrast to the Mendelian inheritance model, parental alleles can contribute unequally to gene expression, which may result in phenotypic variance among individuals and bias in the predicted additive effect of molecular markers associated with production traits. Given the need to understand the effects of allelic variation and parent-of-origin effects on the expression of genes with a commercial interest in cattle, we analyzed the expression of KCNJ11 (potassium inwardly rectifying channel, subfamily J, member 11), which was previously described as a functional candidate gene for meat tenderness. Allele-specific and parent-of-origin-dependent expression of this gene were assessed in bovine muscle using the rs379610823 single nucleotide polymorphism as a reference. Biallelic expression was observed; however, the T allele was expressed at significantly higher levels than the C allele. Furthermore, increased expression of KCNJ11 was found in animals harboring the maternal T allele. This study is the first to describe the differential allelic expression of bovine KCNJ11. Our findings are important for understanding the mechanisms that underlie the pattern of KCNJ11 expression and its potential impact on the phenotypic variation of meat tenderness in Nelore beef cattle. This reinforces the need for further investigation of allelic- and parent-of-origin expression deviation in genetic markers eligible for the selection of target traits.

Implant-Abutment Contact Surfaces and Microgap Measurements of Different Implant Connections Under 3-Dimensional X-Ray Microtomography.

PURPOSE: The presence of a microgap between implant and abutment could produce a bacterial reservoir which could interfere with the long-term health of the periimplant tissues. The aim of this article was to evaluate, by x-ray 3-dimensional microtomography, implant-abutment contact surfaces and microgaps at the implant-abutment interface in different types of implant-abutment connections. MATERIALS AND METHODS: A total of 40 implants were used in this in vitro study. Ten implants presented a screw-retained internal hexagon abutment (group I), 10 had a Morse Cone taper internal connection (group II), 10 another type of Morse Cone taper internal connection (group III), and 10 had a screwed trilobed connection (group IV). RESULTS: In both types of Morse Cone internal connections, there was no detectable separation at the implant-abutment in the area of the conical connection, and there was an absolute congruity without any microgaps between abutment and implant. No line was visible separating the implant and the abutment. On the contrary, in the screwed abutment implants, numerous gaps and voids were present. CONCLUSIONS: The results of this study support the hypothesis that different types of implant-abutment joints are responsible for the observed differences in bacterial penetration.

Genetic identification of a rabies virus from an insectivorous bat in an urban area of Rio Grande do Sul, Brazil.

In Latin America, hematophagous bats are the main reservoirs of rabies virus (RABV) to livestock, to other mammals and, occasionally, to human. Nonetheless, reports of exposure of human and pets to RABV upon aggression by non-hematophagous bats are increasing, possibly facilitated by the synanthropic habits of these bats. We, herein, report the detection and genetic identification of a RABV recovered from an insectivorous bat found sick in a student housing building at the Federal University of Santa Maria, Southern Brazil. Taxonomic characterization identified the captured bat as a member of the genus Nyctinomops, family Molossidae, the group of insectivorous bats. Brain fragments of the bat were positive for RABV antigens by fluorescent antibody test (FAT) and for sequences of the nucleoprotein (N) gene by RT-PCR. The N amplicon was submitted to nucleotide sequencing and analysis, showing that the consensus sequences (SV 33/19) had high identity with RABV sequences of insectivorous bats deposited in GenBank. At phylogenetic tree, the N gene sequences of SV 33/19 clustered with RABV recovered from Nyctinomops laticaudatus, Molossus molossus, and Tadarida lauticaudata bats, and a part of RABV variant 3, 4, and 6, that correspond to Desmodus rotundus, Tadarida brasiliensis, and Lasiurus cinereus, respectively. Although no direct human or domestic animal exposure has been reported, this case strengthens the need for a continuous rabies vaccination in pets in the surrounding areas, since non-hematophagous bats may serve as source of infection for these animals. These findings also call attention for continuous monitoring of populations of synanthropic bats to avoid/prevent human exposure.

Evidence of macrophage modulation in the mouse pubic symphysis remodeling during the end of first pregnancy and postpartum.

In mouse pregnancy, pubic symphysis (PS) remodels into an elastic interpubic ligament (IpL) in a temporally regulated process to provide safe delivery. It restores at postpartum to assure reproductive tract homeostasis. Recently, macrophage localization in the IpL and dynamic changes in the expression of inflammatory mediators observed from the end of pregnancy (D18, D19) to early days postpartum (1dpp, 3dpp) highlighted the necessity of the identification of the key molecules involved in innate immune processes in PS remodeling. Therefore, this study uses morphological and high-sensitivity molecular techniques to identify both macrophage association with extracellular matrix (ECM) remodeling and the immunological processes involved in PS changes from D18 to 3dpp. Results showed macrophage association with active gelatinases and ECM components and 25 differentially expressed genes (DEGs) related to macrophage activities in interpubic tissues from D18 to 3dpp. Additionally, microarray and proteomic analysis showed a significant association of interpubic tissue DEGs with complement system activation and differentially expressed proteins (DEPs) with phagocytosis, highlighting the involvement of macrophage-related activities in mouse PS remodeling. Therefore, the findings suggest that PS ECM remodeling is associated with evidence of macrophage modulation that ensures both IpL relaxation and fast PS recovery postpartum for first labor.

The proton and metal binding sites responsible for the pH-dependent green-red bioluminescence color tuning in firefly luciferases.

Firefly luciferases produce yellow-green light under physiological and alkaline conditions, however at acidic pH, higher temperatures or in the presence of heavy metals the color changes to red, a property called pH-sensitivity. Despite many decades of studies, the proton and metal binding sites responsible for pH-sensitivity remain enigmatic. Previously we suggested that the salt bridge E311/R337 keeps a closed conformation of the luciferin phenolate binding site. Here we further investigated the effect of this salt bridge and mutations of the neighbor residues H310 and E/N354, on metal and pH-sensitivity of firefly luciferases emitting distinct bioluminescence colors (Cratomorphus distinctus: 548 nm; Macrolampis sp2: 569 nm). The substitutions of H310 and E/N354 modulate metal sensitivity, whereas the carboxylate of E311 may work as the catalytic base essential for green bioluminescence and pH-sensitivity. Modeling studies showed that H310, E311 and E354 side-chains coordinate Zinc, constituting the metal binding site and the pH-sensor. Electrostatic potential and pKa calculations suggest that the external couple H310/E354 is affected by pH, whereas E311/R337 make a stabilized internal pair which retains excited oxyluciferin ejected proton near its phenolate group into a high energy state, promoting yellow-green bioluminescence. Protonation or metal binding weaken these electrostatic gates and their ability to retain the excited oxyluciferin released proton near its phenolate, promoting red light emission.

KM+, a mannose-binding lectin from Artocarpus integrifolia: amino acid sequence, predicted tertiary structure, carbohydrate recognition, and analysis of the beta-prism fold.

The complete amino acid sequence of the lectin KM+ from Artocarpus integrifolia (jackfruit), which contains 149 residues/mol, is reported and compared to those of other members of the Moraceae family, particularly that of jacalin, also from jackfruit, with which it shares 52% sequence identity. KM+ presents an acetyl-blocked N-terminus and is not posttranslationally modified by proteolytic cleavage as is the case for jacalin. Rather, it possesses a short, glycine-rich linker that unites the regions homologous to the alpha- and beta-chains of jacalin. The results of homology modeling implicate the linker sequence in sterically impeding rotation of the side chain of Asp141 within the binding site pocket. As a consequence, the aspartic acid is locked into a conformation adequate only for the recognition of equatorial hydroxyl groups on the C4 epimeric center (alpha-D-mannose, alpha-D-glucose, and their derivatives). In contrast, the internal cleavage of the jacalin chain permits free rotation of the homologous aspartic acid, rendering it capable of accepting hydrogen bonds from both possible hydroxyl configurations on C4. We suggest that, together with direct recognition of epimeric hydroxyls and the steric exclusion of disfavored ligands, conformational restriction of the lectin should be considered to be a new mechanism by which selectivity may be built into carbohydrate binding sites. Jacalin and KM+ adopt the beta-prism fold already observed in two unrelated protein families. Despite presenting little or no sequence similarity, an analysis of the beta-prism reveals a canonical feature repeatedly present in all such structures, which is based on six largely hydrophobic residues within a beta-hairpin containing two classic-type beta-bulges. We suggest the term beta-prism motif to describe this feature.

pp-Blast: a "pseudo-parallel" Blast.

We have developed a software called pp-Blast that uses the publicly available Blast package and PVM (parallel virtual machine) to partition a multi-sequence query across a set of nodes with replicated or shared databases. Benchmark tests show that pp-Blast running in a cluster of 14 PCs outperformed conventional Blast running in large servers. In addition, using pp-Blast and the cluster we were able to map all human cDNAs onto the draft of the human genome in less than 6 days. We propose here that the cost/benefit ratio of pp-Blast makes it appropriate for large-scale sequence analysis. The source code and configuration files for pp-Blast are available at http://www.ludwig.org.br/biocomp/tools/pp-blast.

[Percutaneous balloon mitral valvoplasty. Immediate results, complications and hospital outcome]. / Valvoplastia mitral percutânea por balão. Resultados imediatos, complicações e evolução hospitalar.

PURPOSE: To study the short-term results, complications and in-hospital follow-up of 223 percutaneous mitral balloon valvuloplasty (PMBV) procedures (proc)in 219 patients. METHODS: It was used a single 20mm balloon diameter in 4 proc, double balloon in 7, Inoue balloon in 4 and low profile balloon in 196. The mean-age group was 37.19 years. One hundred eighty three (82.1%) procedures were performed in women (mean age, 36.99 years) and 40 (17.9%) in men (mean age, 38.10 years) (p = 0.63). Patients were in functional class II, (NYHA) in 25 (11.2%) procedures, class III in 165 (74.0%) and class IV in 33 (14.8%). Patients were in sinus rhythm in 182 procedures (81.6%) and in atrial fibrillation in 41 (18.4%). The echocardiographic score range from 4 to 14 (7.4% +/- 1.7). Among 4 and 11 were 98.2% of patients. RESULTS: We had 203 complete proc and success, mitral valve area (MVA) > or = 1.5cm2 after PMBV in 194 proc. Echocardiographic MVA before PMBV was 0.9 +/- 0.2cm2 and after 1.8 +/- 0.3cm2 (p < 0.01). Hemodynamic measures MVA before PMBV was 0.9 +/- 0.2cm2 and after was 1.9 +/- 0.3cm2 (p < 0.01). Mean pulmonary artery pressure decreased from 39 +/- 14mmHg to 27 +/- 11mmHg (p < 0.01) and mitral mean gradient from 20 +/- 9mmHg to 6 +/- 5mmHg (p < 0.01). In the 203 proc, mitral valve (MV) was competent in 176 and there were 1+ mitral regurgitation (MR) in 27. After PMBV, MV was competent in 126, and there were 1+ MR in 60, 2+ in 10.3+ in 6 and 4+ MR in 1. There was complication in 15 proc, severe MR in 7 (3 or 4+), stroke in 3 and cardiac tamponade in 5. Two patients died during emergency cardiac surgery after left ventricular perforation and one by stroke. CONCLUSION: PMBV was an effective procedure with a high grade of success and low rate of complication.

[Identification of intracavitary electrograms predictive of success in ablation of anomalous pathways by radiofrequency]. / Identificação dos electrogramas intracavitários preditivos de sucesso na ablação de vias anômalas por radiofreqüência.

PURPOSE: To analyze the characteristics of atrial (A) and ventricular (V) electrograms, AV and VA intervals relation to determine a precise radiofrequency ablation site. METHODS: A and V electrograms (EG) recorded at successful (group 1) and unsuccessful (group 2) sites in 22 patients submitted to RF catheter ablation were identified. Specific criteria as morphology, AV and VA intervals relation and the presence of an accessory pathway were analyzed. RESULTS: The shortest AV interval with a VEG that precedes the delta wave in the onset of the QRS recorded at the EKG leads was the best outcome predictor. The AV connection potential was only recorded in five out of 22 patients and did not interfere with the result when not present. The interval measured between the onset of the V electrogram and the onset (delta wave) of the QRS complex (V-d) varied from -4.9 to -11msec (m = 7.5 + 1.6) in G1 and from -6 to 15msec (m = 9.6 +/- 3) in G2 (p = 0.03). Unsuccessful sites had the VEG preceding the delta wave but the AV interval was not short. Success in the retrograde mapping of target site seems to be the presence of an atrial potential that occurs simultaneously with the nadir (S wave) of the QRS complex. CONCLUSION: Specific criteria such as morphology of the local VEG, a short AV or VA intervals and a VEG that precede the onset (delta wave) of the QRS identified a successful site for RF ablation. When present the accessory pathway potential indicates a high chance of a successful ablation; however when not present did not decrease the chance of success.

[Tricuspid catheter-balloon valvuloplasty. Report of 4 cases]. / Valvoplastia tricúspide por cateter balão. Relato de quatro casos.

Tricuspid stenosis was treated in four patients by percutaneous balloon valvotomy. A mean pressure gradient equal or higher than 3 mmHg across the tricuspid valve using Doppler echocardiography and the increase of this pressure gradient during inspiration were the most significant criteria for diagnosis and quantification of tricuspid stenosis. There was a considerable hemodynamic improvement with reduction of the tricuspid valve gradient and relief of symptoms after balloon tricuspid valvotomy.

[Angioplasty of distal aorta and iliac, femoral, popliteal, renal and subclavian arteries. Early and late results]. / Angioplastia de aorta distal e artérias ilíaca, femoral, poplítea, renal e subclávia. Resultados imediatos e tardios.

PURPOSE: To study the late results of peripheral angioplasty. PATIENTS AND METHODS: In the period of 8 years from August, 1981 until August, 1989, 27 patients were submitted to 33 procedures of peripheral angioplasty. RESULTS: There was success in 29 procedures, an insufficient dilatation in 1 and and failure in 3 (2 new attempts were effective). Success rate was 88% of the procedures; clinical and angiographic success was reached in 25 (93%) of the 27 patients. Thirty four obstructions were successful dilated: 12 in renal artery, 12 in common iliac artery, 4 in external iliac artery, 3 in superficial femoral artery, 1 in distal aorta. In the evolution we had a restenosis of a renal artery that was redilated, a precocious occlusion of a common iliac artery (9% of common iliac artery dilatations and 6% of the total of the iliac dilatations) and a popliteal occlusion. Of the 34 dilatations we had a patency of 91% until 2 months. CONCLUSION: Angioplasty showed to be an effective method with good results in the long term follow-up.

[Percutaneous mitral valvuloplasty with the single balloon technique. Short-term results, complications and in-hospital follow-up]. / Valvoplastia mitral percutânea com a técnica do balão único. Resultados imediatos, complicações e evolução intra-hospitalar.

PURPOSE: To study the short-term results, complication and in-hospital follow-up of 268 percutaneous mitral balloon valvuloplasty (PMBV) procedures performed with the low-profile monofoil balloon (LPMB) technique from 1990 to 1995. METHODS: A single 30mm balloon diameter was used in 247 (92.9%) procedures, a single 25mm balloon diameter in 9 (3.3%), a single 25mm balloon followed by a single 30mm balloon diameter in 7 (2.6%) and in 5 procedures a balloon was not used. The mean age group was 36 +/- 12 years. Two hundred nineteen (81.7%) procedures were performed in women (mean age 36 +/- 12 years) and 49 (18.3%) in men (mean age, 35 +/- 14 years) (p = 0.78). Patients were in functional class II (NYHA) in 39 (14.5%), class III in 198 (73.9%) and class IV in 31 (11.6%). Patients were in sinus rhythm in 228 (85.1%) procedures and in atrial fibrillation in 40 (14.9%). The echocardiographic score ranged from 4 to 14 (mean 7.2 +/- 1.5). RESULTS: There were 256 complete procedures, 249 of which were successful (mitral valve area (MVA) > or = 1.5cm2 after PMBV). Echocardiographic calculated MVA before PMBV was 0.9 +/- 0.2cm2. Hemodynamic calculated MVA before PMBV was 0.9 +/- 0.2cm2 and after was 2.0 +/- 0.4cm2 (p < 0.000001). Mean pulmonary artery pressure decreased from 40 +/- 15mmHg to 28 +/- 10mmHg (p < 0.000001) and mitral mean gradient from 20 +/- 7mmHg to 5 +/- 4mmHg (p < 0.000001). In the 256 complete procedures mitral valve (MV) was competent in 214 and there was 1+ mitral regurgitation (MR) in 42. After PMBV, MV was competent in 166 and there was 1+ MR in 68, 2+ in 16, 3+ in 5 and 4+ MR in 1. There were complications in 14 (5.2%) procedures, severe MR in 6 (3 or 4+), stroke in 2 and cardiac tamponade in 6. Two patients died during emergency cardiac surgery after left ventricular perforation and 1 after stroke. CONCLUSION: PMBV with the LPMB was an effective procedure with a high success rate and a low rate of complications as the more usual double-balloon and Inoue balloon techniques.

[Single balloon versus Inoue balloon in percutaneous mitral balloon valvuloplasty. Short-term results and complications]. / Balãco Unico versus balão de Inoue na valvoplastia mitral percutânea por balão. Resultados imediatos e complicações.

PURPOSE: To assess short-term results and complications of percutaneous mitral balloon valvuloplasty (PMBV) performed with Inoue balloon (IB) and single low profile balloon (SB). METHODS: We performed 390 PMBV procedures, 29 with IB and 337 with SB. There were no differences in age, sex, echocardiographic score distribution and echocardiographic mitral valve area (MVA). RESULTS: We performed 29 complete procedures with IB and 330 of 337 in SB group. Comparing IB and pre and pos-PMBV data we obtained: mean pulmonary artery pressure (MPAP) 36 +/- 15 and 39 +/- 14 mmHg, p = 0.2033, mean mitral gradient 17 +/- 6 and 20 +/- 77 mmHg, p = 0.0396 and MVA 0.9 +/- 0.2 and 0.9 +/- 0.2 cm2, p = 0.8043 and pos-PMBV:MPAP 25 +/- 8 and 28 +/- 10 mmHg, p = 0.2881, gradient 5 +/- 3 and 5 +/- 4 mmHg, p = 0.2778 and MVA 2.2 +/- 0.2 and 2.0 +/- 0.4 cm2, p = 0.0362. Mitral valve (MV) was competent in 26 patients in IB and in 280 in SB group and we had +/4 mitral regurgitation in 3 patients in IB and in 57 in SB group (p = 0.3591) pre-PMBV respectively and pos-PMBV there was also no difference in MV competence (p = 0.7439). CONCLUSION: Both techniques were effective. Hemodynamic data were also similar although MVA was greater in IB group after PMBV.

Hyperbaric oxygen therapy induces kidney protection in an ischemia/reperfusion model in rats.

Ischemia/reperfusion (I/R) injury remains a major cause of graft dysfunction, which impacts short- and long-term follow-up. Hyperbaric oxygen therapy (HBO), through plasma oxygen transport, has been currently used as an alternative treatment for ischemic tissues. The aim of this study was to analyze the effects of HBO on kidney I/R injury model in rats, in reducing the harmful effect of I/R. The renal I/R model was obtained by occluding bilateral renal pedicles with nontraumatic vascular clamps for 45 minutes, followed by 48 hours of reperfusion. HBO therapy was delivered an hypebaric chamber (2.5 atmospheres absolute). Animals underwent two sessions of 60 minutes each at 6 hours and 20 hours after initiation of reperfusion. Male Wistar rats (n = 38) were randomized into four groups: sham, sham operated rats; Sham+HBO, sham operated rats exposed to HBO; I/R, animals submitted to I/R; and I/R+HBO, I/R rats exposed to HBO. Blood, urine, and kidney tissue were collected for biochemical, histologic, and immunohistochemical analyses. The histopathological evaluation of the ischemic injury used a grading scale of 0 to 4. HBO attenuated renal dysfunction after ischemia characterized by a significant decrease in blood urea nitrogen (BUN), serum creatinine, and proteinuria in the I/R+HBO group compared with I/R alone. In parallel, tubular function was improved resulting in significantly lower fractional excretions of sodium and potassium. Kidney sections from the I/R plus HBO group showed significantly lower acute kidney injury scores compared with the I/R group. HBO treatment significantly diminished proliferative activity in I/R (P < .05). There was no significant difference in macrophage infiltration or hemoxygenase-1 expression. In conclusion, HBO attenuated renal dysfunction in a kidney I/R injury model with a decrease in BUN, serum creatinine, proteinuria, and fractional excretion of sodium and potassium, associated with reduced histological damage.