Actividad antimicrobiana de weissella confusa y sus metabolitos frente a escherichia coli y klebsiella pneumoniae

Título en ingles: Antimicrobial activity of Weissella confuse and its metabolites against Escherichia coli and  Klebsiella pneumoniaeResumen: Con el fin evaluar el campo de aplicación potencial de una bacteria ácido láctica y de sus metabolitos, se realizó la cinética de la actividad antimicrobiana de W. confusa y de sus  metabolitos contra E. coli, y K. pneumoniae, dos patógenos causantes de enfermedades transmitidas por alimentos. La producción de W. confusa se realizó por fermentación discontinua en sustrato comercial MRS. Se realizaron tres fermentaciones durante 6 horas, sin aireación, agitación continúa 33°C y 100 rpm. Cada hora de fermentación se separaron tres sustancias biológicas, W. confusa con sus metabolitos (W+W10b), células de W. confusa libres de metabolitos (W) y metabolito (W10b) y se midió la actividad antimicrobiana contra los patógenos E. coli, y K. pneumoniae. Se encontraron diferencias estadísticas significativas entre tratamientos y tiempo de fermentación. Para E. coli el tratamiento W presentó la mayor actividad antimicrobiana, la cual se obtuvo entre la cuarta y sexta hora de fermentación (2.45 cm de diámetro promedio de inhibición). Para K. pneumoniae, los tratamientos W y W+W10b presentaron actividad antimicrobiana entre la cuarta y quinta hora de fermentación, sin diferencia significativa entre ellos. W. confusa y el metabolito W10b demostraron poseer capacidad antimicrobiana contra E. coli y K. pneumoniae, lo cual sugiere que W. confusa y W10b podrían utilizarse como alternativa de bioconservación o bioprotección de alimentos frescos y procesados, para alimentación humana y animal; y podría convertirse en una alternativa al uso de antibióticos para enfermedades causadas por E. coli y K. pneumoniae.Palabras clave: bioconservación, alimentos, enfermedades, aplicaciones biotecnológicas.Summary: The kinetic of antimicrobial activity ofWeissella confusa and their metabolites against E. coli, and K. pneumoniae, (two pathogens causing foodborne illness) was evaluated, in order to know the possible use in food processing. W. confusa was produced by batch fermentation using MRS commercial substrate. Three fermentations, of 6 hours at 33 ° C, without aeration, stirring continuously (100 rpm) were performed. In every hour of fermentation, three biological substances, W. confusa with their metabolites (W + W10b), W. confusa free cells metabolites (W), and metabolite (W10b) were separated, and subsequently the antimicrobial activity against pathogenic E. coli and K. pneumoniae was measured. Statistically significant differences between treatments and fermentation time were found. Treatment (W) against E. coli, showed the greatest antimicrobial activity, it was obtained between the fourth and the sixth hours of fermentation (2.45 cm inhibition diameter average). In treatments W and W + W10b against K. pneumoniae, statistically significant differences between them were not found. The antimicrobial activity was shown between the fourth and fifth hour of fermentation. W. confusa and W10b have antimicrobial activity against E. coli andK. pneumoniae, suggesting that W and W10b could be used as an alternative to biopreservation or bioprotection of fresh and processed food for human and animal consumption, and could become an alternative to antibiotics used for diseases caused by E. coli and K. pneumoniae.Key words: bioconservation, food, diseases, biotechnology applications

Impact of COVID-19 on cardiovascular testing in the United States versus the rest of the world

Objectives: This study sought to quantify and compare the decline in volumes of cardiovascular procedures between the United States and non-US institutions during the early phase of the coronavirus disease-2019 (COVID-19) pandemic. Background: The COVID-19 pandemic has disrupted the care of many non-COVID-19 illnesses. Reductions in diagnostic cardiovascular testing around the world have led to concerns over the implications of reduced testing for cardiovascular disease (CVD) morbidity and mortality. Methods: Data were submitted to the INCAPS-COVID (International Atomic Energy Agency Non-Invasive Cardiology Protocols Study of COVID-19), a multinational registry comprising 909 institutions in 108 countries (including 155 facilities in 40 U.S. states), assessing the impact of the COVID-19 pandemic on volumes of diagnostic cardiovascular procedures. Data were obtained for April 2020 and compared with volumes of baseline procedures from March 2019. We compared laboratory characteristics, practices, and procedure volumes between U.S. and non-U.S. facilities and between U.S. geographic regions and identified factors associated with volume reduction in the United States. Results: Reductions in the volumes of procedures in the United States were similar to those in non-U.S. facilities (68% vs. 63%, respectively; p = 0.237), although U.S. facilities reported greater reductions in invasive coronary angiography (69% vs. 53%, respectively; p < 0.001). Significantly more U.S. facilities reported increased use of telehealth and patient screening measures than non-U.S. facilities, such as temperature checks, symptom screenings, and COVID-19 testing. Reductions in volumes of procedures differed between U.S. regions, with larger declines observed in the Northeast (76%) and Midwest (74%) than in the South (62%) and West (44%). Prevalence of COVID-19, staff redeployments, outpatient centers, and urban centers were associated with greater reductions in volume in U.S. facilities in a multivariable analysis. Conclusions: We observed marked reductions in U.S. cardiovascular testing in the early phase of the pandemic and significant variability between U.S. regions. The association between reductions of volumes and COVID-19 prevalence in the United States highlighted the need for proactive efforts to maintain access to cardiovascular testing in areas most affected by outbreaks of COVID-19 infection

Polimorfismo e expressão de genes candidatos e suas relações com o crescimento e as características da carne em bovinos nelore (Bos indicus)

Com o intuito de estudar o potencial de aplicação de genes candidatos na seleção animal para o melhoramento genético da qualidade de carne de bovinos Nelore (Bos indicus), o presente trabalho teve como objetivos principais: (1) Avaliar em bovinos Nelore e cruzamentos Nelore x Bos taurus a ocorrência de associações entre o SNP g.98535683A>G:BTAU7 do gene CAST e as características da carne produzida e (2) quantificar a expressão gênica e proteica da cadeia pesada da miosina em bovinos Nelore com características fenotípicas contrastantes de crescimento e maciez da carne. Para o desenvolvimento do primeiro objetivo, 500 animais foram genotipados para o SNP g.98535683A>G:BTAU7 e fenotipados para força de cisalhamento (FC), índice de fragmentação miofibrilar (MFI), área de olho de lombo (AOL), espessura de gordura subcutânea (EGS) e lipídeos totais (LIP). Associação significativa entre o SNP e a maciez da carne foi observada. O genótipo AA apresentou os melhores valores em relação a esta característica. As diferenças encontradas entre os genótipos AA e AG (AA - AG) foram de - 0,19 kg e 5,31 para FC e MFI, respectivamente. O SNP não mostrou associação significativa com AOL, EGS e LIP. Para a execução do segundo objetivo, 90 bovinos Nelore terminados em confinamento foram utilizados. Levando em consideração o peso final ao abate (PF) e a FC dos 90 animais, 24 indivíduos com características contrastantes de peso e maciez da carne foram selecionados e divididos em quatro grupos experimentais (6 animais/grupo). Os grupos corresponderam a animais leves com carne macia (PF = 493,50 kg e FC = 3,76 kg), animais leves com carne dura (PF = 515,66 kg e FC = 8,06 kg), animais pesados com carne macia (PF = 604,33 kg e FC = 3,99) e animais pesados com carne dura (PF = 605,33 kg e FC = 7,83 kg). Em cada grupo, foram avaliadas características fenotípicas como MFI, AOL, EGS, LIP, índice de marmorização (IM), coloração instrumental (L*, a*, b*) e perdas por cozimento (PT). A análise de expressão dos genes MYH7, MYH2 e MYH1 foi feita por PCR em tempo real (RT-qPCR). A caracterização e quantificação proteica das isoformas da MyHC (MyHC-I, MyHC-IIa e MyHC-IIx/d) foi realizada por eletroforese SDS-PAGE. Em todas as análises (qualidade da carne e expressão gênica e proteica) não foi observada interação significativa entre crescimento e maciez da carne. Nas características AOL, EGS, IM e LIP não foi observado efeito significativo do crescimento e a maciez da carne. Porém, os animais pesados apresentaram maiores valores de L*, a* e b* em relação aos animais leves. Na maciez, o grupo de carne macia mostrou maiores valores de L* (32,29 ± 2,78) que o grupo de carne dura (29,10 ± 2,61). Por outro lado, maiores PT foram observadas em animais com carne dura em relação a animais com carne macia. Nos resultados de expressão gênica, o gene MYH7 não mostrou diferenças entre animais leves e pesados. Entretanto, a expressão dos genes MYH2 e MYH1 foi significativamente menor nos animais pesados em relação aos animais leves. Quanto à maciez da carne, a expressão dos três genes não diferiram significativamente entre grupos de carne dura e macia. Na expressão proteica, as três isoformas (MyHC-I, IIa e IIx), não mostraram diferença significativa entre animais leves e pesados. No entanto, as proporções da MyHC-I foram significativamente maiores no grupo de carne dura (16,51 ± 5,47) em relação ao grupo de carne macia (9,12 ± 3,16) e pelo contrário, as proporções da MyHC-IIa foram maiores no grupo de carne macia (73,78 ± 4,45) em relação ao grupo de carne dura (65,45 ± 8,79). A isoforma MyHC-IIx/d não mostrou efeito significativo sobre a maciez. Os resultados deste trabalho mostraram o potencial de aplicação do SNP g.98535683A>G:BTAU7 do gene CAST e da isoforma MyHC-IIa na seleção de animais para o melhoramento da maciez da carne de bovinos Nelore, além de, manifestar o diferencial de expressão dos genes MYH1 e MYH2 entre animais pesados e leves.In order to study the potential of application of candidate genes in animal selection for breeding meat of Nellore cattle, this work had as main objectives: (1) Assess in Nellore cattle (Bos indicus) and crosses Nellore x Bos taurus the occurrence of associations between the g.98535683A>G:BTAU7 SNP in the CAST gene and traits of the meat produced and (2) quantify the gene and protein expression of myosin heavy chain in Nellore cattle with contrasting phenotypic traits of growth and meat tenderness. For the development of first objective, 500 animals were genotyped for the g.98535683A>G:BTAU7 SNP and phenotyped for shear force (SF), myofibrillar fragmentation index (MFI), ribeye area (RA), backfat thickness (BT) and total lipids (TL). Significant association between the SNP and meat tenderness was observed. The AA genotype showed the best values in relation to this trait. The differences found between genotypes AA and AG (AA - AG) were – 0.19 kg and 5.31 for SF and MFI, respectively. The SNP did not show significant association with RA, BT and TL. For the execution of the second aim, 90 feedlot Nellore cattle were used. Considering the final slaughter weight (FW) and the SF of the 90 animals, 24 individuals with contrasting characteristics of weight and tenderness of meat were selected and divided into four experimental groups (6 animals / group). The groups corresponded to lightweight animals with tender meat (FW = 493.50 kg and SF = 3.76 kg), lightweight animals with tough meat (FW = 515.66 kg and SF = 8.06 kg), heavy animals with tender meat (FW = 604.33 kg and SF = 3.99) and heavy animals with tough meat (FW = 605.33 kg and SF = 7.83 kg). In each group, were evaluated phenotypic characteristics as MFI, RA, BT, TL, marbling índex (MI), instrumental color (L*, a*, b*) and cooking losses (CL). The expression analysis of the MYH7, MYH2 and MYH1 genes was performed by Real-Time PCR (RT-qPCR). The characterization and quantification of the MyHC isoforms (MyHC-I, MyHC-IIa and MyHC-IIx/d) was carried out by SDS-PAGE. In all analyzes (meat quality and gene and protein expression) there was no significant interaction between growth and meat tenderness. In the traits MFI, RA, BT, IM and TL it was not observed significant effect of the growth and the tenderness of meat. However, the heavy animals showed higher values of L*, a* and b* than lightweight animals. In the tenderness, the tender meat group showed higher values of the L* (32.29 ± 2.78) than the tough meat group (29.10 ± 2.61). On the other hand, higher CL were observed in animals with tough meat than animals with tender meat. In the results of gene expression, the MYH7 gene did not show differences between lightweight and heavy animals. However, the expression of the MYH2 and MYH1 genes was significantly lower in heavy animals than lightweight animals. As for meat tenderness, the expression of the three genes did not differ significantly between tough and tender meat groups. In the protein expression, the three isoforms (MHC-I, IIa and IIx), did not show significant difference between lightweight and heavy animals. Nevertheless, the MyHC-I proportions were significantly higher in the tough meat group (16.51 ± 5.47) than tender meat group (9.12 ± 3.16) and on the contrary, the proportions of the MyHC-IIa were higher in tender meat group (73.78 ± 4.45) than tough meat group (65.45 ± 8.79). The MyHC-IIx isoform did not show significant effect on tenderness meat. The results of this work showed the potential of application of the g.98535683A>G:BTAU7 SNP in gene CAST and the MyHC-IIa isoform in animal selection to improve meat tenderness of Nellore cattle, in addition, it showed the differential expression of MYH1 and MYH2 genes between heavy and lightweight animals

Actividad antimicrobiana de weissella confusa y sus metabolitos frente a escherichia coli y klebsiella pneumoniae

Título en ingles: Antimicrobial activity of Weissella confuse and its metabolites against Escherichia coli and  Klebsiella pneumoniaeResumen: Con el fin evaluar el campo de aplicación potencial de una bacteria ácido láctica y de sus metabolitos, se realizó la cinética de la actividad antimicrobiana de W. confusa y de sus  metabolitos contra E. coli, y K. pneumoniae, dos patógenos causantes de enfermedades transmitidas por alimentos. La producción de W. confusa se realizó por fermentación discontinua en sustrato comercial MRS. Se realizaron tres fermentaciones durante 6 horas, sin aireación, agitación continúa 33°C y 100 rpm. Cada hora de fermentación se separaron tres sustancias biológicas, W. confusa con sus metabolitos (W+W10b), células de W. confusa libres de metabolitos (W) y metabolito (W10b) y se midió la actividad antimicrobiana contra los patógenos E. coli, y K. pneumoniae. Se encontraron diferencias estadísticas significativas entre tratamientos y tiempo de fermentación. Para E. coli el tratamiento W presentó la mayor actividad antimicrobiana, la cual se obtuvo entre la cuarta y sexta hora de fermentación (2.45 cm de diámetro promedio de inhibición). Para K. pneumoniae, los tratamientos W y W+W10b presentaron actividad antimicrobiana entre la cuarta y quinta hora de fermentación, sin diferencia significativa entre ellos. W. confusa y el metabolito W10b demostraron poseer capacidad antimicrobiana contra E. coli y K. pneumoniae, lo cual sugiere que W. confusa y W10b podrían utilizarse como alternativa de bioconservación o bioprotección de alimentos frescos y procesados, para alimentación humana y animal; y podría convertirse en una alternativa al uso de antibióticos para enfermedades causadas por E. coli y K. pneumoniae.Palabras clave: bioconservación, alimentos, enfermedades, aplicaciones biotecnológicas.Summary: The kinetic of antimicrobial activity ofWeissella confusa and their metabolites against E. coli, and K. pneumoniae, (two pathogens causing foodborne illness) was evaluated, in order to know the possible use in food processing. W. confusa was produced by batch fermentation using MRS commercial substrate. Three fermentations, of 6 hours at 33 ° C, without aeration, stirring continuously (100 rpm) were performed. In every hour of fermentation, three biological substances, W. confusa with their metabolites (W + W10b), W. confusa free cells metabolites (W), and metabolite (W10b) were separated, and subsequently the antimicrobial activity against pathogenic E. coli and K. pneumoniae was measured. Statistically significant differences between treatments and fermentation time were found. Treatment (W) against E. coli, showed the greatest antimicrobial activity, it was obtained between the fourth and the sixth hours of fermentation (2.45 cm inhibition diameter average). In treatments W and W + W10b against K. pneumoniae, statistically significant differences between them were not found. The antimicrobial activity was shown between the fourth and fifth hour of fermentation. W. confusa and W10b have antimicrobial activity against E. coli andK. pneumoniae, suggesting that W and W10b could be used as an alternative to biopreservation or bioprotection of fresh and processed food for human and animal consumption, and could become an alternative to antibiotics used for diseases caused by E. coli and K. pneumoniae.Key words: bioconservation, food, diseases, biotechnology applications

Gene and protein expression of myosin heavy chain in Nellore cattle comparing growth or meat tenderness traits

The objective was to investigate gene and protein expression of myosin heavy chain (MyHC) in Nellore cattle slaughtered at different weights (BW) or degrees of meat tenderness. Ninety animals with initial BW 370 ± 37 kg, 24 months of age, were slaughtered after 95 days on feed. We evaluated shear force (SF), myofibrillar fragmentation index, ribeye area, backfat thickness, marbling, color, and cooking losses. Subsequently, 24 animals were selected and divided into four contrasting groups, in which light (BW ¼ 504.58 ± 32.36 kg) versus heavy animals (BW ¼ 604.83 ± 42.97 kg) and animals with tender (SF ¼ 3.88 ± 0.57 kg) versus tough meat (SF¼ 7.95 ± 1.04 kg) were compared. The MYH7, MYH2 and MYH1 genes were analyzed by real-time PCR. The MyHC isoforms (MyHC-I, MyHC-IIa, and MyHC-IIx) were quantified by SDS-PAGE electrophoresis. We found lower expression of MYH2 and MYH1 genes in heavy compared to light animals and a higher amount of MyHC-I isoform in the tough meat group compared to the tender meat group. Protein expression of MyHC-IIa was higher in the tender meat group. A negative correlation was found of this protein and SF (tenderness), suggesting MyHC-IIa as a biomarker of meat quality

Application of proteomic to investigate the different degrees of meat tenderness in Nellore breed

This study describes the association between meat tenderness and abundance of soluble muscle proteins in Nellore bulls (Bos indicus) using a proteomic approach. We evaluated shear force (SF) of Longissimus thoracis muscle 24 h after slaughter and selected three experimental groups of animals with moderately tender (TE; SF = 3.9 ± 0.7 kg), moderately tough (TO; SF = 5.6 ± 0.7 kg) and very tough meat (TO+; SF = 7.9 ± 1.4 kg). Proteome was investigated by two-dimensional electrophoresis (2D-PAGE) in combination with electrospray ionization-tandem mass spectrometry (ESI–MS/MS). The metabolic proteins triosephosphate isomerase (TPI1) and phosphoglucomutase 1 (PGM1), the structural protein profilin 1 (PFN1), and cytosol aminopeptidase (LAP3) were up-regulated (P < 0.05) in the TE meat group when compared to the TO and TO+ groups. Actin structural proteins (ACTA1, ACTB, and ACTG1), the oxidative stress protein peroxiredoxin (PRDX6, PRDX2, PRDX1, and PARK7), heat shock protein isoforms, and co-chaperones (CDC37 and STIP1) were up-regulated (P < 0.05) in the TO and TO+ meat groups. In addition, we also identified proteins PFN1, LAP3, PRDX1, PRDX2, HSPD1, and ARHGDIA to be associated with beef tenderness. The results reported herein demonstrated that meat tenderness in Nellore cattle depends on the modulation and expression of a set of proteins involved in different biological pathways.Incluye referencias bibliográfica

Stoma-free survival after anastomotic leak following rectal cancer resection: worldwide cohort of 2470 patients

Background: The optimal treatment of anastomotic leak after rectal cancer resection is unclear. This worldwide cohort study aimed to provide an overview of four treatment strategies applied. Methods: Patients from 216 centres and 45 countries with anastomotic leak after rectal cancer resection between 2014 and 2018 were included. Treatment was categorized as salvage surgery, faecal diversion with passive or active (vacuum) drainage, and no primary/secondary faecal diversion. The primary outcome was 1-year stoma-free survival. In addition, passive and active drainage were compared using propensity score matching (2: 1). Results: Of 2470 evaluable patients, 388 (16.0 per cent) underwent salvage surgery, 1524 (62.0 per cent) passive drainage, 278 (11.0 per cent) active drainage, and 280 (11.0 per cent) had no faecal diversion. One-year stoma-free survival rates were 13.7, 48.3, 48.2, and 65.4 per cent respectively. Propensity score matching resulted in 556 patients with passive and 278 with active drainage. There was no statistically significant difference between these groups in 1-year stoma-free survival (OR 0.95, 95 per cent c.i. 0.66 to 1.33), with a risk difference of -1.1 (95 per cent c.i. -9.0 to 7.0) per cent. After active drainage, more patients required secondary salvage surgery (OR 2.32, 1.49 to 3.59), prolonged hospital admission (an additional 6 (95 per cent c.i. 2 to 10) days), and ICU admission (OR 1.41, 1.02 to 1.94). Mean duration of leak healing did not differ significantly (an additional 12 (-28 to 52) days). Conclusion: Primary salvage surgery or omission of faecal diversion likely correspond to the most severe and least severe leaks respectively. In patients with diverted leaks, stoma-free survival did not differ statistically between passive and active drainage, although the increased risk of secondary salvage surgery and ICU admission suggests residual confounding

Stoma-free Survival After Rectal Cancer Resection With Anastomotic Leakage: Development and Validation of a Prediction Model in a Large International Cohort.

Objective:To develop and validate a prediction model (STOMA score) for 1-year stoma-free survival in patients with rectal cancer (RC) with anastomotic leakage (AL).Background:AL after RC resection often results in a permanent stoma.Methods:This international retrospective cohort study (TENTACLE-Rectum) encompassed 216 participating centres and included patients who developed AL after RC surgery between 2014 and 2018. Clinically relevant predictors for 1-year stoma-free survival were included in uni and multivariable logistic regression models. The STOMA score was developed and internally validated in a cohort of patients operated between 2014 and 2017, with subsequent temporal validation in a 2018 cohort. The discriminative power and calibration of the models' performance were evaluated.Results:This study included 2499 patients with AL, 1954 in the development cohort and 545 in the validation cohort. Baseline characteristics were comparable. One-year stoma-free survival was 45.0% in the development cohort and 43.7% in the validation cohort. The following predictors were included in the STOMA score: sex, age, American Society of Anestesiologist classification, body mass index, clinical M-disease, neoadjuvant therapy, abdominal and transanal approach, primary defunctioning stoma, multivisceral resection, clinical setting in which AL was diagnosed, postoperative day of AL diagnosis, abdominal contamination, anastomotic defect circumference, bowel wall ischemia, anastomotic fistula, retraction, and reactivation leakage. The STOMA score showed good discrimination and calibration (c-index: 0.71, 95% CI: 0.66-0.76).Conclusions:The STOMA score consists of 18 clinically relevant factors and estimates the individual risk for 1-year stoma-free survival in patients with AL after RC surgery, which may improve patient counseling and give guidance when analyzing the efficacy of different treatment strategies in future studies

Search for supersymmetry with a compressed mass spectrum in the vector boson fusion topology with 1-lepton and 0-lepton final states in proton-proton collisions at s=\sqrt{s}= 13 TeV

A search for supersymmetric particles produced in the vector boson fusion topology in proton-proton collisions is presented. The search targets final states with one or zero leptons, large missing transverse momentum, and two jets with a large separation in rapidity. The data sample corresponds to an integrated luminosity of 35.9 fb$^{-1}$ of proton-proton collisions at $\sqrt{s}=$ 13 TeV collected in 2016 with the CMS detector at the LHC. The observed dijet invariant mass and lepton-neutrino transverse mass spectra are found to be consistent with the standard model predictions. Upper limits are set on the cross sections for chargino ($\widetilde\chi_1^\pm$) and neutralino ($\widetilde\chi_2^0$) production with two associated jets. For a compressed mass spectrum scenario in which the $\widetilde\chi_1^\pm$ and $\widetilde\chi_2^0$ decays proceed via a light slepton and the mass difference between the lightest neutralino $\widetilde\chi_1^0$ and the mass-degenerate particles $\widetilde\chi_1^\pm$ and $\widetilde\chi_2^0$ is 1 (30) GeV, the most stringent lower limit to date of 112 (215) GeV is set on the mass of these latter two particles