A variant associated to IGF-1 mRNA and protein expression in sheep.

The insulin-like growth factor-1 (IGF-1), a key hormone in muscle development was investigated for single-nucleotide polymorphisms (SNPs) upstream of the IGF-1 gene and their effects upon its cognate mRNA and hormone levels in sheep. A 70 d feeding trial was conducted with 22 F1 (Dorper × Pelibuey) lambs, individually allocated and fed a diet with a forage-to-concentrate ratio of 36:64 and 17% crude protein. Sequence analyses of 265 bp upstream the IGF-1 gene revealed the variant NC_040254.1:g.[184028491G > C;184028493G > A]. These SNPs generate alleles A and B, with frequencies of 0.66 and 0.34 in F1 lambs and of 0.73 and 0.27 in 81 pure Dorper lambs, respectively. Females were grouped by genotype AA, AB and BB (n = 3). IGF-1 hormone concentrations at 14, 42 and 70 d were higher (p < 0.05) in AA lambs compared to AB + BB lambs. The IGF-1 mRNA level was 2.6-fold higher in AA animals (n = 5, p < 0.05) than in AB + BB lambs (n = 7). A DNA binding site for the Inhibitor of Growth family member 4 (ING4) was found in allele B but not in allele A, which could explain the lower mRNA and hormone expression levels for AB + BB animals. The variant reported here appears to function as an eQTL with a negative effect on the level of IGF-1.

Integrative proteomics and phosphoproteomics reveals phosphorylation networks involved in the maintenance and expression of embryogenic competence in sugarcane callus.

Plant embryogenic cell culture allows mass propagation and genetic manipulation, but the mechanisms that determine the fate of these totipotent cells in somatic embryos have not yet been elucidated. Here, we performed label-free quantitative proteomics and phosphoproteomics analyses to determine signaling events related to sugarcane somatic embryo differentiation, especially those related to protein phosphorylation. Embryogenic calli were compared at multiplication (EC0, dedifferentiated cells) and after 14 days of maturation (EC14, onset of embryo differentiation). Metabolic pathway analysis showed enriched lysine degradation and starch/sucrose metabolism proteins during multiplication, whereas the differentiation of somatic embryos was found to involve the enrichment of energy metabolism, including the TCA cycle and oxidative phosphorylation. Multiplication-related phosphoproteins were associated with transcriptional regulation, including SNF1 kinase homolog 10 (KIN10), SEUSS (SEU), and LEUNIG_HOMOLOG (LUH). The regulation of multiple light harvesting complex photosystem II proteins and phytochrome interacting factor 3-LIKE 5 were predicted to promote bioenergetic metabolism and carbon fixation during the maturation stage. A motif analysis revealed 15 phosphorylation motifs. The [D-pS/T-x-D] motif was overrepresented during somatic embryo differentiation. A protein-protein network analysis predicted interactions among SNF1-related protein kinase 2 (SnRK2), abscisic acid-responsive element-binding factor 2 (ABF2), and KIN10, which indicated the role of these proteins in embryogenic competence. The predicted interactions between TOPLESS (TPL) and histone deacetylase 19 (HD19) may be involved in posttranslational protein regulation during somatic embryo differentiation. These results reveal the protein regulation dynamics of somatic embryogenesis and new players in somatic embryo differentiation, including their predicted phosphorylation motifs and phosphosites.

In vitro assessment of two novel Cellulases from Trabulsiella odontotermitis for agricultural waste utilization.

BACKGROUND: The production of agricultural wastes still growing as a consequence of the population growing. However, the majority of these residues are under-utilized due their chemical composition, which is mainly composed by cellulose. Actually, the search of cellulases with high efficiency to degrade this carbohydrate remains as the challenge. In the present experiment, two genes encoding an endoglucanase (EC 3.2.1.4) and ß-glucosidase (EC 3.2.1.21) were overexpressed in Escherichia coli and their recombinant enzymes (egl-FZYE and cel-FZYE, respectively) characterized. Those genes were found in Trabulsiella odontermitis which was isolated from the gut of termite Heterotermes sp. Additionally, the capability to release sugars from agricultural wastes was evaluated in both enzymes, alone and in combination. RESULTS: The results have shown that optimal pH was 6.0 and 6.5, reaching an activity of 1051.65 ± 47.78 and 607.80 ± 10.19 U/mg at 39 °C, for egl-FZYE and cel-FZYE, respectively. The Km and Vmax for egl-FZYE using CMC as substrate were 11.25 mg/mL and 3921.57 U/mg, respectively, whereas using Avicel were 15.39 mg/mL and 2314.81 U/mg, respectively. The Km and Vmax for cel-FZYE using Avicel as substrate were 11.49 mg/mL and 2105.26 U/mg, respectively, whereas using CMC the enzyme did not had activity. Both enzymes had effect on agricultural wastes, and their effect was improved when they were combined reaching an activity of 955.1 ± 116.1, 4016.8 ± 332 and 1124.2 ± 241 U/mg on corn stover, sorghum stover and pine sawdust, respectively. CONCLUSIONS: Both enzymes were capable of degrading agricultural wastes, and their effectiveness was improved up to 60% of glucose released when combined. In summary, the results of the study demonstrate that the recombinant enzymes exhibit characteristics that indicate their value as potential feed additives and that the enzymes could be used to enhance the degradation of cellulose in the poor-quality forage generally used in ruminant feedstuffs.

Label-Free Quantitative Phosphoproteomics Reveals Signaling Dynamics Involved in Embryogenic Competence Acquisition in Sugarcane.

In this study, a label-free quantitative phosphoproteomic analysis was performed to identify and quantify signaling events related to the acquisition of embryogenic competence in sugarcane. Embryogenic and nonembryogenic calli were compared at the multiplication phase, resulting in the identification of 163 phosphoproteins unique to embryogenic calli, 9 unique to nonembryogenic calli, and 51 upregulated and 40 downregulated in embryogenic calli compared to nonembryogenic calli. Data are available via ProteomeXchange with identifier PXD018054. Motif-x analysis revealed the enrichment of [xxxpSPxxx], [RxxpSxxx], and [xxxpSDxxx] motifs, which are predicted phosphorylation sites for several kinases related to stress responses. The embryogenic-related phosphoproteins (those unique and upregulated in embryogenic calli) identified in the present study are related to abscisic acid-induced signaling and abiotic stress response; they include OSK3, ABF1, LEAs, and RD29Bs. On the other hand, the nonembryogenic-related phosphoproteins EDR1 and PP2Ac-2 are negative regulators of abscisic acid signaling, suggesting a relationship between phosphoproteins involved in the abscisic acid and stress responses in the acquisition of embryogenic competence. Moreover, embryogenic-related phosphoproteins associated with epigenetic modifications, such as HDA6, HDA19, and TOPLESS, and with RNA metabolism, including AGO1, DEAH5, SCL30, UB2C, and SR45, were identified to play potential roles in embryogenic competence. These results reveal novel phosphorylation sites for several proteins and identify potential candidate biomarkers for the acquisition of embryogenic competence in sugarcane.

Salmonella enterica Optimizes Metabolism After Addition of Acyl-Homoserine Lactone Under Anaerobic Conditions.

Acyl-homoserine lactones (AHLs) are quorum sensing (QS) signaling molecules that mediate cell-to-cell communication in Gram-negative bacteria. Salmonella does not produce AHL, however, it can recognize AHLs produced by other species through SdiA protein modulating important cellular functions. In this work, the influence of the N-dodecanoyl-DL-homoserine lactone (C12-HSL) on glucose consumption, metabolic profile, and gene expression of Salmonella throughout the cultivation time in Tryptic Soy Broth (TSB) under anaerobic conditions was evaluated. Analysis of the supernatant culture in high-performance liquid chromatography (HPLC) revealed lower glucose uptake after 4 and 6 h of the addition of C12-HSL. Gas chromatography-mass spectrometry (GC-MS) based analysis of the intracellular metabolites revealed C12-HSL perturbation in the abundance levels of metabolites related to the metabolic pathways of glycerolipids, purines, amino acids, and aminoacyl-tRNA biosynthesis. The real-time quantitative PCR (RT-qPCR) indicated that Salmonella increase expression of genes associated with nucleoside degradation and quantification of metabolites supported the induction of pentose phosphate pathway to ensure growth under lower glucose consumption. The obtained data suggest an important role of C12-HSL in the optimization of metabolism at a situation of high population densities.

Bioactive Properties of Syzygium cumini (L.) Skeels Pulp and Seed Phenolic Extracts.

The emergence of bacterial strains resistant to different antibiotics has prompted the search for new sources of antimicrobial compounds. Studies have shown that jambolan [Syzygium cumini (L.) Skeels], a tropical fruit from the Mirtaceae family, contains a great variety of phytochemical compounds with high antioxidant and antimicrobial activity. This study aimed to determine the centesimal composition and physicochemical characteristics of the pulp and seed of S. cumini (L.) Skeels, as well as the content of total phenolic compounds and the antioxidant, antibacterial, antibiofilm and anti-quorum sensing (QS) activities of the phenolic extracts obtained from the pulp and the seeds of this fruit. The in vitro antibacterial and anti-QS activities of active films incorporating phenolic extracts were also evaluated. Additionally, we performed molecular docking of phenolic compounds present in jambolan with the CviR QS regulator of Chromobacterium violaceum. The composition and physicochemical characteristics of the samples presented similar values to those found for the species. However, the seed phenolic extract had a higher content of phenolic compounds and antioxidant activity than the pulp. Both phenolic extracts presented antibacterial activity against Aeromonas hydrophila, C. violaceum, Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica serovar Typhimurium, Serratia marcescens, Listeria monocytogenes, and Staphylococcus aureus. The seed phenolic extract was particularly inhibitory against S. aureus. The pulp phenolic extract inhibited swarming motility and biofilm formation of A. hydrophila, E. coli, and S. marcescens in sub-MIC concentrations. The pulp and seed phenolic extracts inhibited violacein production in C. violaceum. Films incorporating both phenolic extracts inhibited the growth of bacteria, particularly Pseudomonas fluorescens, L. monocytogenes, and S. aureus, as well as QS in C. violaceum. Molecular docking showed that a variety of compounds found in pulp and seed extracts of jambolan, particularly chlorogenic acid and dihydroquercetin, potentially bind CviR protein and may interfere with QS. Our results indicate that pulp and seed of jambolan are good sources of antibacterial, antibiofilm, and anti-QS compounds that can be used in the development of natural preservatives and for application in antibacterial active films.

Influence of genetic markers on the feeding behavior of yearling bulls / Influencia de los marcadores genéticos en el comportamiento alimentario de toretes de un año / Influência de marcadores genéticos no comportamento alimentar de touros de um ano

Abstract Background: The search for gene and marker effects on economically important traits is aimed not only to understanding the genetic architecture of complex traits but also to applying the information to breeding schemes. Objective: To analyze the effect of two temperament-related SNPs (rs109576799 located in the DRD3 gene, and rs43696138 in the HTR2A gene) on feeding performance of Mexican beef cattle. Methods: One hundred and thirty-six young beef bulls were included in a centralized feed efficiency performance test based on residual feed intake (RFI), with 20 d for adaptation and 70 d of feed efficiency testing. In addition to feeding traits, temperament was assessed at the beginning of the trial using pen score (PS) and exit velocity (EV). All animals were genotyped with two markers located in the HTR2A and DRD3 genes, and an association analysis was conducted between these genotypes and the measured traits. Results: For Brangus breed, a significant association was obtained between average daily gain (ADG; p=0.019), and the rs43696138 marker, resulting in higher gains for homozygous genotype GG (1.69 ± 0.04 kg), when compared to the heterozygous genotype GA (1.54 ± 0.04 kg). Conclusion: The previously reported association of these markers with temperament was not confirmed in the evaluated breeds; however, the rs43696138 marker showed an effect on a feeding performance trait. Further studies are needed to determine the effect of this and other markers on both RFI and temperament.

Resumen Antecedentes: La búsqueda de efectos genéticos y marcadores de rasgos económicamente relevantes no solo se basa en el interés biológico de comprender la arquitectura genética de rasgos complejos, sino también en aplicar la información en los esquemas prácticos de mejoramiento. Objetivo: Analizar el efecto de dos SNPs relacionados con el temperamento (rs109576799 localizado en el gen DRD3, y rs43696138 localizado en el gen HTR2A) sobre la eficiencia alimenticia en el ganado bovino mexicano. Métodos: Ciento treinta y seis toretes de carne jóvenes fueron sometidos a una prueba de comportamiento alimenticio basada en el consumo residual de alimento (RFI), con 20 d de adaptación y 70 d de prueba para la eficiencia alimenticia. Además de los rasgos de comportamiento alimenticio, se evaluó el temperamento de los animales al inicio de la prueba, mediante la evaluacion de comportamiento en el corral (PS), y la velocidad de salida (EV). Todas las muestras se tipificaron con dos marcadores localizados en los genes DRD3 y HTR2A para posteriormente realizar un análisis de asociación de los genotipos con los rasgos evaluados. Resultados: En la raza Brangus, se observó una asociación significativa de la media de ganancia diaria de peso (ADG, p=0,019) con el marcador rs43696138, localizado en el gen HTR2A, resultando en mayores ganancias para el genotipo GG (1,69 ± 0,04 kg) en comparación con los toros heterocigóticos GA (1,54 ± 0,04 kg). Conclusión: No se confirmó la asociación de estos marcadores previamente reportados con el temperamento en las razas evaluadas; sin embargo, el marcador rs43696138 mostró efecto en un rasgo de comportamiento alimenticio. Se necesitan más estudios para determinar el efecto de éste y otros marcadores en el consumo residual de alimento (RFI) y el temperamento.

Resumo Antecedentes: A busca de efeitos genéticos e marcadores de características economicamente relevantes não se baseia apenas no interesse biológico de compreender a arquitetura genética de traços complexos, mas também na aplicação da informação nos esquemas práticos de melhoria. Objetivo: Analisar o efeito de dois SNPs relacionados ao temperamento (rs109576799 localizado no gene DRD3 e rs43696138 localizado no gene HTR2A) sobre a eficiência nutricional no gado mexicano. Métodos: Cento e trinta e seis touros jovens foram submetidos a um teste de comportamento alimentar com base na entrada de alimentação residual (RFI), com 20 d de adaptação e 70 d de teste para eficiência de alimentação. Além dos traços de comportamento alimentar, o temperamento dos animais foi avaliado no início do teste, através da avaliação do comportamento na caneta (PS) e da velocidade de saída (EV). Todas as amostras foram digitadas com dois marcadores localizados nos genes DRD3 e HTR2A para posteriormente realizar uma análise de associação dos genótipos com os traços avaliados. Resultados: Na raça Brangus, observou-se uma associação significativa do ganho diário médio (ADG, p = 0,019) com o marcador rs43696138, localizado no gene HTR2A, resultando em maiores ganhos para o genótipo GG (1,69 ± 0,04 kg), em comparação com os touros heterozigóticos GA (1,54 ± 0,04 kg). Conclusão: A associação destes marcadores previamente relatados com o temperamento nas raças avaliadas não foi confirmada; no entanto, o marcador rs43696138 mostrou um efeito sobre uma característica de comportamento alimentar. Mais estudos são necessários para determinar o efeito deste e outros marcadores com ingestão alimentar residual (RFI) e temperamento.

In vitro reduction of methane production by 3-nitro-1-propionic acid is dose-dependent1.

Methanogenesis is a metabolic process that allows the rumen ecosystem the ability to maintain the low hydrogen partial pressures needed for proper digestive function. However, rumen methanogenesis is considered to be an inefficient process because it can result in the loss of 4% to 12% of the total energy consumed by the host. Recent studies have shown that some short-chain nitrocompounds such as nitroethane, 2-nitroethanol, 2-nitro-1-propanol, and 3-nitro-1-propionic acid (3NPA) are capable of inhibiting the production of methane during in vitro culture; nevertheless, optimal supplementation doses have yet to be determined. In the present study, in vitro cultures of freshly collected mixed populations of ruminal microbes were supplemented with the naturally occurring nitrocompound, 3NPA, to achieve 0, 3, 6, 9, or 12 mM. Analysis of fermentation products after 24 h of incubation revealed that methane (CH4) production was reduced in a dose-dependent manner by 29% to 96% (P < 0.05) compared with the amount produced by untreated controls (15.03 ± 0.88 µmol mL-1 incubated liquid). Main effects of the supplement were also observed, which resulted in a reduction (P < 0.05) on amounts of total gas and volatile fatty acids (VFA) produced, as well as in an increase of 0.07 to 0.30 µmol mL-1 on rates of 3NPA degradation. Changes in production of metabolites as CH4, hydrogen (H2), VFA, and NH3 indicated that the fermentation efficiency was not compromised dramatically by 3NPA treatment in moderate doses of 6 and 9 mM. Results further revealed that the metabolism of the 3NPA by microbial populations is also dose-dependent. The microbes were able to metabolize more than 75% of the added nitrocompound, with the greatest degradation rates in cultures treated with 9-mM 3NPA. Finally, from a practical standpoint, and considering the magnitude of CH4 reduction, effect on VFA, and percentage of metabolized supplement, the most efficacious dose for 3NPA administration may be between 3 and 9 mM.

Draft Genome Sequences for Five Strains of Trabulsiella odontotermitis, Isolated from Heterotermes sp. Termite Gut.

Trabulsiella odontotermitis represents a novel species in the genus Trabulsiella with no complete genome reported yet. Here, we describe the draft genome sequences of five isolates from termites present in the north of Mexico, which have an interesting pool of genes related to cellulose degradation with biotechnological application.

Genome Sequence of Citrobacter sp. CtB7.12, Isolated from the Gut of the Desert Subterranean Termite Heterotermes aureus.

The draft genome of Citrobacter sp. CtB7.12, isolated from termite gut, is presented here. This organism has been reported as a cellulolytic bacterium, which is biotechnologically important because it can be used as a gene donor for the ethanol and biofuel industries.

Hepatectomia laparoscópica no tratamento das metástases hepáticas / Laparoscopic hepatectomy for liver metastasis

Descreve-se experiência inicial e resultados imediatos da hepatectomia laparoscópica no tratamento de metástases hepáticas em quatro pacientes. Uma paciente foi submetida a hepatectomia direita e três foram submetidas a bissegmentectomia 6-7. O tempo médio cirúrgico foi de 360 minutos e 4 dias de internação. Uma paciente apresentou fístula biliar autolimitada. Mortalidade operatória foi nula. Conclui-se que hepatectomia maior laparoscópica é factível, segura e apresenta excelentes resultados imediatos, com retorno precoce às atividades profissionais. Entretanto, ainda são necessários estudos com maior número de casos e trabalhos comparativos para se estabelecer limites e indicações deste procedimento em metástases hepáticas.

The authors describe their initial experience and immediate results with laparoscopic liver resection in the treatment of four patients with liver metastasis. Three patients underwent laparoscopic anatomic bisegmentectomy 6-7 and one laparoscopic right hemihepatectomy. Mean operation time was 360 minutes and median hospital stay was 4 days. One patient developed bile leakage that was treated conservatively. No postoperative mortality was observed. This initial experience demonstrates that laparoscopic liver resection can be safely done with excellent immediate results. However larger and comparative studies are still needed to establish indications and limits for laparoscopy in the treatment of liver metastasis.

[Laparoscopic hepatectomy for liver metastasis]. / Hepatectomia laparoscópica no tratamento das metástases hepáticas.

The authors describe their initial experience and immediate results with laparoscopic liver resection in the treatment of four patients with liver metastasis. Three patients underwent laparoscopic anatomic bisegmentectomy 6-7 and one laparoscopic right hemihepatectomy. Mean operation time was 360 minutes and median hospital stay was 4 days. One patient developed bile leakage that was treated conservatively. No postoperative mortality was observed. This initial experience demonstrates that laparoscopic liver resection can be safely done with excellent immediate results. However larger and comparative studies are still needed to establish indications and limits for laparoscopy in the treatment of liver metastasis.