Essential oils from Argentinean native species reduce in vitro methane production

The aim of this study was to evaluate the effect of Argentinean essential oils (EO) on methane production during in vitro fermentation compared to EO with proven effects as rumen fermentation modifiers. A complete randomized block design was used and the treatments included EO from Aloysia gratissima (50, 100, 150 and 300 mg/L), Eucalyptus globulus (5, 50, 150 and 300 mg/L), Lippia turbinata (30, 60, 120 and 240 mg/L), Mentha x piperita var. vulgaris (50, 100, 200 and 400 mg/L), Origanum vulgare ssp. hirtum cv. Compacto (0.5, 5, 50 and 250 mg/L) and cv. Mendocino (50, 150, 250 and 350 mg/L), Rosmarinus officinalis (100, 300, 500 and 700 mg/L), Schinus molle (75, 150, 300 and 600 mg/L), Tagetes minuta (5, 50, 125 and 250 mg/L), and Thymus vulgaris (5, 50, 150 and 300 mg/L). Two controls were included: control (not containing EO) and monensin (1.87 mg/L). Variables measured were: digestibility of neutral detergent fiber, gas and methane production. Compared to the control, monensin reduced methane production by 44%, but with a 15% reduction in fiber digestibility. Except for E. globulus and L. turbinata, each variable measured was significantly reduced with the higher level of inclusion compared to the control, representing that overall microbial activity was affected. L. turbinata (60.8% limonene) reduced methane by half compared to control, and by 35 to 85% when compared to monensin, without affecting digestibility of the fiber. Some essential oils from native Argentinean plants exhibited great potential to reduce enteric methane production without affecting digestibility, of which L. turbinata was the most promising alternative.El objetivo de este estudio fue evaluar el efecto de aceites esenciales (AE) de plantas nativas de Argentina en la producción de metano durante la fermentación in vitro, en comparación con AE con efectos comprobados como modificadores de la fermentación ruminal. Se realizó un diseño en bloque completos al azar y los AE y dosis evaluados fueron: Aloysia gratissima (0,5, 5, 50 y 250 mg/L) , Eucalyptus globulus (5, 50, 150 y 300 mg/L), Lippia turbinata (30, 60, 120 y 240 mg/L), Mentha x piperita var. vulgaris (50, 100, 200 y 400 mg/L), Origanum vulgare ssp. hirtum cv. Compacto (0,5, 5, 50 y 250 mg/L) y cv. Mendocino (50, 150, 250 y 350 mg/L), Rosmarinus officinalis (100, 300, 500 y 700 mg/L), Schinus molle (75, 150, 300 y 600 mg/L), Tagetes minuta (5, 50, 125 y 250 mg/L), y Thymus vulgaris (5, 50, 150 y 300 mg/L). Se incluyeron dos controles: control (sin AE) y monensina (1,87 mg/L). Las variables evaluadas fueron: digestibilidad in vitro de la fibra en detergente neutro, producción de gas y metano. En comparación al control, la monensina redujo la producción de metano en un 44%, pero con una reducción de un 15% en la digestibilidad de la fibra. Con excepción a E. globulus y L. turbinata, todas las variables fueron reducidas significativamente con el mayor nivel de inclusión comparado con el control, lo cual representa que la actividad microbiana se vio afectada. L. turbinata (60,8% limoneno) redujo la producción de metano a la mitad comparado al control, y entre un 35 y 85% en comparación a la monensina, sin que la digestibilidad de la fibra se vea afectada. Algunos aceites esenciales de plantas nativas de Argentina demostraron gran potencial para reducir la producción de metano entérico sin afectar la digestibilidad, de los cuales L. turbinata fue la alternativa más promisoria.EEA ManfrediFil: García, F. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; ArgentinaFil: Brunetti, Maria Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Manfredi; ArgentinaFil: Lucini, Enrique I. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias,; ArgentinaFil: Scorcione Turcato, M.C. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Moreno, Maria Valeria. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Manfredi; ArgentinaFil: Frossasco, Georgina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Manfredi; ArgentinaFil: Colombatto, Dario. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Agronomia; Argentina.Fil: Martinez, Maria Jose. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Manfredi; ArgentinaFil: Martinez Ferrer, Jorge. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Manfredi; Argentin

KPC-2 allelic variants in Klebsiella pneumoniae isolates resistant to ceftazidime-avibactam from Argentina: blaKPC-80, blaKPC-81, blaKPC-96 and blaKPC-97

Ceftazidime-avibactam (CZA) therapy has significantly improved survival rates for patients infected by carbapenem-resistant bacteria, including KPC producers. However, resistance to CZA is a growing concern, attributed to multiple mechanisms. In this study, we characterized four clinical CZA-resistant Klebsiella pneumoniae isolates obtained between July 2019 and December 2020. These isolates expressed novel allelic variants of blaKPC-2 resulting from changes in hotspots of the mature protein, particularly in loops surrounding the active site of KPC. Notably, KPC-80 had an K269_D270insPNK mutation near the Lys270-loop, KPC-81 had a del_I173 mutation within the Ω-loop, KPC-96 showed a Y241N substitution within the Val240-loop and KPC-97 had an V277_I278insNSEAV mutation within the Lys270-loop. Three of the four isolates exhibited low-level resistance to imipenem (4 µg/mL), while all remained susceptible to mero meropenem. Avibactam and relebactam effectively restored carbapenem susceptibility in resistant isolates. Cloning mutant blaKPC genes into pMBLe increased imipenem MICs in recipient Escherichia coli TOP10 for blaKPC-80, blaKPC-96, and blaKPC-97 by two dilutions; again, these MICs were restored by avibactam and relebactam. Frameshift mutations disrupted ompK35 in three isolates. Additional resistance genes, including blaTEM-1, blaOXA-18 and blaOXA-1, were also identified. Interestingly, three isolates belonged to clonal complex 11 (ST258 and ST11) and one to ST629. This study highlights the emergence of CZA resistance including unique allelic variants of blaKPC-2 and impermeability. Comprehensive epidemiological surveillance and in-depth molecular studies are imperative for understanding and monitoring these complex resistance mechanisms, crucial for effective antimicrobial treatment strategies.Fil: Sanz, María Belén. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Pasteran, Fernando. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: de Mendieta, Juan Manuel. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Brunetti, Florencia Lourdes. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Investigaciones en Bacteriología y Virología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Albornoz, Ezequiel Pablo. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Rapoport, Melina. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Lucero, Celeste. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Errecalde, Laura. Gobierno de la Ciudad de Buenos Aires. Hospital General de Agudos "Juan A. Fernández"; ArgentinaFil: Nuñez, Maria Rosa. Provincia del Neuquen. Hospital Provincial Neuquen "dr. E. Castro Rendon"; ArgentinaFil: Monge, Renata. Hospital Británico de Buenos Aires; ArgentinaFil: Pennini, Magdalena Ines. Laboratorio Stamboulian; ArgentinaFil: Power, Pablo. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Corso, Alejandra. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; ArgentinaFil: Gómez, Sonia Alejandra. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

The copy number variation landscape of congenital anomalies of the kidney and urinary tract

Congenital anomalies of the kidney and urinary tract (CAKUT) are a major cause of pediatric kidney failure. We performed a genome-wide analysis of copy number variants (CNVs) in 2,824 cases and 21,498 controls. Affected individuals carried a significant burden of rare exonic (that is, affecting coding regions) CNVs and were enriched for known genomic disorders (GD). Kidney anomaly (KA) cases were most enriched for exonic CNVs, encompassing GD-CNVs and novel deletions; obstructive uropathy (OU) had a lower CNV burden and an intermediate prevalence of GD-CNVs; and vesicoureteral reflux (VUR) had the fewest GD-CNVs but was enriched for novel exonic CNVs, particularly duplications. Six loci (1q21, 4p16.1-p16.3, 16p11.2, 16p13.11, 17q12 and 22q11.2) accounted for 65% of patients with GD-CNVs. Deletions at 17q12, 4p16.1-p16.3 and 22q11.2 were specific for KA; the 16p11.2 locus showed extensive pleiotropy. Using a multidisciplinary approach, we identified TBX6 as a driver for the CAKUT subphenotypes in the 16p11.2 microdeletion syndrome

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field