Clonal diversity of Acinetobacter clinical isolates producing NDM-type carbapenemase in Cuba, 2013-19.

Objectives: Our study aimed to elucidate the clonal diversity of carbapenem-resistant Acinetobacter clinical isolates producing NDM-type carbapenemase collected through national surveillance in Cuba during a 7-year period (2013-19). Methods: A total of 199 isolates of Acinetobacter spp. from 37 hospitals in 12 provinces were genetically analyzed for their species, carbapenemase genes and genotypes. Sequence type (ST) and OXA-51-like gene type were determined for bla NDM-positive isolates. Results: Most isolates (95%) were identified as species of Acinetobacter calcoaceticus-baumannii complex, with A. baumannii being the majority. Acquired carbapenemase genes were assigned to bla OXA or bla NDM type; the most commonly detected gene was bla OXA-23-like (49%), followed by bla OXA-24-like (20%) and bla NDM (15%). Twenty-nine bla NDM-positive isolates (22 A. baumannii, 2 A. pittii, 2 A. johnsonii, 3 other species) were differentiated into 19 STs, including the most common, ST23. Though NDM genes were mostly typed as bla NDM-1, a novel bla NDM-42 was identified in an ST79 A. baumannii isolate. bla OXA-51-like genes of NDM-positive A. baumannii were discriminated into 10 OXA types, including 2 novel ones. Conclusions: Our study indicated the spread of bla NDM to various clones of A. baumannii and other Acinetobacter spp. in Cuba.

Asociación de polimorfismos en los genes CAPN y CAST con propiedades fisicoquímicas de la carne bovina: una revisión / Association of polymorphisms in CAPN and CAST genes with physicochemical properties of beef: a review / Associação de polimorfismos nos genes CAPN e CAST com propriedades físico-químicas da carne bovina: uma revisão

Resumen Las propiedades fisicoquímicas como terneza, color, jugosidad, sabor, marmoleo, pH y capacidad de retención de agua, son consideradas como los atributos que tienen mayor importancia en la calidad de la carne. El manejo post mortem influye en el funcionamiento del sistema proteolítico de las enzimas µ-calpaínas y calpastatinas, que están codificadas por los genes CAPN1 y CAST, respectivamente. Esta revisión muestra las asociaciones de los polimorfismos CAPN1 y CAST con el fin de explicar las propiedades fisicoquímicas antes mencionadas, que determinan las características relacionadas con la calidad de la carne. Existen polimorfismos de CAPN1 y CAST asociados a propiedades fisicoquímicas particulares de la carne, pero no todos los polimorfismos se asocian con las propiedades mencionadas. La actividad del sistema proteolítico de las enzimas varía según el manejo peri y post mortem. La presencia o ausencia de un polimorfismo varía entre razas. Por consiguiente, las asociaciones de polimorfismos de nucleótido simple con las propiedades fisicoquímicas de la carne mencionadas en esta revisión podrían ayudar a mejorar los indicadores de calidad de la carne bovina.

Abstract Physicochemical properties such as tenderness, color, juiciness, flavor, marbling, pH and water retention capacity, are considered the most important attributes in the meat quality. Posmortem management influences the functioning of the proteolytic system of the enzymes calpains and calpastatins, which are encoded by CAPN1 and CAST genes, respectively. This review displays the associations of CAPN1 and CAST polymorphisms in order to explain the aforementioned physicochemical properties, which determine the characteristics relate to the beef quality. There are CAPN1 and CAST polymorphisms associated with particular physicochemical properties of meat, but not all polymorphisms are associated with the mentioned properties. The activity of enzymes proteolytic system varies according to peri and posmortem management. The presence or absence of a polymorphism varies between races. Therefore, the associations of single nucleotide polymorphisms with the physicochemical properties of meat mentioned in this review could help to improve the quality indicators of beef.

Resumo Propriedades físico-químicas como maciez, cor, suculência, sabor, marmorização, pH e capacidade de retenção da água, são consideradas os atributos mais importantes na qualidade da carne. O manejo pós-morte influencia o funcionamento do sistema proteolítico das enzimas calpaína e calpastatina, que são codificadas pelos genes CAPN1 e CAST, respectivamente. Esta revisão mostra as associações dos polimorfismos CAPN1 e CAST para explicar as propriedades físico-químicas citadas, que determinam as características relacionadas à qualidade da carne. Existem polimorfismos CAPN1 e CAST associados a propriedades físico-químicas específicas da carne, mas nem todos os polimorfismos estão associados às propriedades mencionadas. A atividade das enzimas do sistema proteolítico varia de acordo com o manejo peri e pos-mortem. A presença ou ausência de um polimorfismo varia entre as raças. Portanto, as associações de polimorfismos de nucleotídeo único com as propriedades físico-químicas da carne mencionadas nesta revisão poderiam ajudar a melhorar os indicadores de qualidade da carne bovina.

Influência da insensibilização inadequada durante o abate de suínos nas propriedades físico-químicas da carne / Influence of inadequate sttuning during swine slaughter on the meat physical-chemical properties

Objetivou-se avaliar a influência da insensibilização inadequada durante o abate de suínos na cor, pH e perdas de água da carne. Após eletrocussão dos animais em um abatedouro-frigorífico sob SIF da região da Zona da Mata, Minas Gerais, parâmetros de insensibilização foram avaliados em 3.347 suínos abatidos. Os parâmetros avaliados foram ausência de reflexos palpebrais, reflexo de orelhas, gritos, pedalagem nos membros anteriores e respiração arrítmica. Após o completo abate e 16 horas de estocagem das carcaças em câmara fria a 2°C, fragmentos transversais do músculo Longissimus dorsi foram obtidos de animais selecionados de forma randomizada e insensibilizados de forma adequada (n=12) e inadequada (n=12). As amostras de carne foram avaliadas quanto ao pH, Capacidade de Retenção de Água (CRA). Do total de 3.347 suínos abatidos, 272 (8,13%) apresentaram pelo menos algum sinal de insensibilização inadequada. Observou-se que as 24 amostras de carne tiveram pH médio de 6,06, perda média de 24,74% de água e escore médio de 3,64 na escala de cor avaliada. O pH médio da carne obtida de animais inadequadamente insensibilizados (6,11) foi maior (p<0,05) que o pH médio de animais adequadamente insensibilizados (6,01). Quanto menor o pH da carne menor era o seu escore de cor e CRA (p<0,05). Portanto, a carne obtida de suínos mal insensibilizados possui maior pH, o que resulta em carne mais escura e com baixa CRA.(AU)

The objective of study was to evaluate the influence of inadequate sttuning during the swine slaughter in color, pH, and water loss of meat. After electric sttuning of animals in a slaugtherhouse under SIF of the Zona da Mata region, Minas Gerais, stunning parameters were evaluated in 3,347 slaughtered swines. The parameters evaluated were the absence of eyelid reflexes, ear reflexes, screams, pedaling in the forelimbs and arrhythmic breathing. After the full 16 hours of slaughter and storage of carcases in a cold chamber at 2°C, transverse fragments Longissimus dorsi muscle were obtained from randomly selected animals and properly (n=12) and improperly (n=12) sttuninged animals. The meat samples were evaluated for pH, Water Holding Capacity (WHC). Of the 3,347 slaughtered swines, 272 (8.13%) showed at least some sign of inadequate sttuning. It was observed that the 24 meat samples had mean pH of 6.06, mean loss water of 24.74% and mean score of 3.64 evaluated in color scale. The mean pH of meat obtained from animals improperly sttuninged (6.11) was greater (p<0.05) than the mean pH of animal properly sttuninged (6.01). The lower pH of meat causes lower color score and WHC (p<0.05). Therefore, the meat obtained from improperly sttuninged swine has a higher pH, which results in darker meat and low WHC.(AU)

Influência da insensibilização inadequada durante o abate de suínos nas propriedades físico-químicas da carne / Influence of inadequate sttuning during swine slaughter on the meat physical-chemical properties

Objetivou-se avaliar a influência da insensibilização inadequada durante o abate de suínos na cor, pH e perdas de água da carne. Após eletrocussão dos animais em um abatedouro-frigorífico sob SIF da região da Zona da Mata, Minas Gerais, parâmetros de insensibilização foram avaliados em 3.347 suínos abatidos. Os parâmetros avaliados foram ausência de reflexos palpebrais, reflexo de orelhas, gritos, pedalagem nos membros anteriores e respiração arrítmica. Após o completo abate e 16 horas de estocagem das carcaças em câmara fria a 2°C, fragmentos transversais do músculo Longissimus dorsi foram obtidos de animais selecionados de forma randomizada e insensibilizados de forma adequada (n=12) e inadequada (n=12). As amostras de carne foram avaliadas quanto ao pH, Capacidade de Retenção de Água (CRA). Do total de 3.347 suínos abatidos, 272 (8,13%) apresentaram pelo menos algum sinal de insensibilização inadequada. Observou-se que as 24 amostras de carne tiveram pH médio de 6,06, perda média de 24,74% de água e escore médio de 3,64 na escala de cor avaliada. O pH médio da carne obtida de animais inadequadamente insensibilizados (6,11) foi maior (p<0,05) que o pH médio de animais adequadamente insensibilizados (6,01). Quanto menor o pH da carne menor era o seu escore de cor e CRA (p<0,05). Portanto, a carne obtida de suínos mal insensibilizados possui maior pH, o que resulta em carne mais escura e com baixa CRA.

The objective of study was to evaluate the influence of inadequate sttuning during the swine slaughter in color, pH, and water loss of meat. After electric sttuning of animals in a slaugtherhouse under SIF of the Zona da Mata region, Minas Gerais, stunning parameters were evaluated in 3,347 slaughtered swines. The parameters evaluated were the absence of eyelid reflexes, ear reflexes, screams, pedaling in the forelimbs and arrhythmic breathing. After the full 16 hours of slaughter and storage of carcases in a cold chamber at 2°C, transverse fragments Longissimus dorsi muscle were obtained from randomly selected animals and properly (n=12) and improperly (n=12) sttuninged animals. The meat samples were evaluated for pH, Water Holding Capacity (WHC). Of the 3,347 slaughtered swines, 272 (8.13%) showed at least some sign of inadequate sttuning. It was observed that the 24 meat samples had mean pH of 6.06, mean loss water of 24.74% and mean score of 3.64 evaluated in color scale. The mean pH of meat obtained from animals improperly sttuninged (6.11) was greater (p<0.05) than the mean pH of animal properly sttuninged (6.01). The lower pH of meat causes lower color score and WHC (p<0.05). Therefore, the meat obtained from improperly sttuninged swine has a higher pH, which results in darker meat and low WHC.

Avaliação de propriedades funcionais e oxidação lipídica em mortadela com aplicação de maca peruana / Evaluation of functional properties and lipid oxidation in mortadella with application of maca peruana

Avaliou-se durante 90 dias o pH, a cor, a capacidade de retenção de água (CRA), a textura e a oxidação lipídica, de uma formulação de mortadela otimizada por meio de um delineamento experimental de mistura para dois componentes. Desenvolveram-se duas amostras da formulação de mortadela otimizada, com antioxidante (OA) e sem antioxidante (OS). Os valores de pH e CRA não apresentaram diferenças significativas ao final dos 90 dias entre as amostras, OA (6,02 e 94,05 g/100g) e OS (6,07 e 93,25 g/100g). Na cor houve diferenças significativas. Na textura, apenas a dureza, mastigabilidade e coesividade apresentaram diferenças significativas. E aos 90 dias as amostras com (2,10 mg de MDA/Kg) e sem antioxidante (2,59 mg de MDA/Kg) não apresentaram diferenças significativas para oxidação lipídica e podem ser considerados aceitáveis para produtos cárneos.(AU)

Avaliação de propriedades funcionais e oxidação lipídica em mortadela com aplicação de maca peruana / Evaluation of functional properties and lipid oxidation in mortadella with application of maca peruana

Avaliou-se durante 90 dias o pH, a cor, a capacidade de retenção de água (CRA), a textura e a oxidação lipídica, de uma formulação de mortadela otimizada por meio de um delineamento experimental de mistura para dois componentes. Desenvolveram-se duas amostras da formulação de mortadela otimizada, com antioxidante (OA) e sem antioxidante (OS). Os valores de pH e CRA não apresentaram diferenças significativas ao final dos 90 dias entre as amostras, OA (6,02 e 94,05 g/100g) e OS (6,07 e 93,25 g/100g). Na cor houve diferenças significativas. Na textura, apenas a dureza, mastigabilidade e coesividade apresentaram diferenças significativas. E aos 90 dias as amostras com (2,10 mg de MDA/Kg) e sem antioxidante (2,59 mg de MDA/Kg) não apresentaram diferenças significativas para oxidação lipídica e podem ser considerados aceitáveis para produtos cárneos.

Microbial Cell Factories à la Carte: Elimination of Global Regulators Cra and ArcA Generates Metabolic Backgrounds Suitable for the Synthesis of Bioproducts in Escherichia coli.

Manipulation of global regulators is one of the strategies used for the construction of bacterial strains suitable for the synthesis of bioproducts. However, the pleiotropic effects of these regulators can vary under different conditions and are often strain dependent. This study analyzed the effects of ArcA, CreC, Cra, and Rob using single deletion mutants of the well-characterized and completely sequenced Escherichia coli strain BW25113. Comparison of the effects of each regulator on the synthesis of major extracellular metabolites, tolerance to several compounds, and synthesis of native and nonnative bioproducts under different growth conditions allowed the discrimination of the particular phenotypes that can be attributed to the individual mutants and singled out Cra and ArcA as the regulators with the most important effects on bacterial metabolism. These data were used to identify the most suitable backgrounds for the synthesis of the reduced bioproducts succinate and 1,3-propanediol (1,3-PDO). The Δcra mutant was further modified to enhance succinate synthesis by the addition of enzymes that increase NADH and CO2 availability, achieving an 80% increase compared to the parental strain. Production of 1,3-PDO in the ΔarcA mutant was optimized by overexpression of PhaP, which increased more than twice the amount of the diol compared to the wild type in a semidefined medium using glycerol, resulting in 24 g · liter-1 of 1,3-PDO after 48 h, with a volumetric productivity of 0.5 g · liter-1 h-1IMPORTANCE Although the effects of many global regulators, especially ArcA and Cra, have been studied in Escherichia coli, the metabolic changes caused by the absence of global regulators have been observed to differ between strains. This scenario complicates the identification of the individual effects of the regulators, which is essential for the design of metabolic engineering strategies. The genome of Escherichia coli BW25113 has been completely sequenced and does not contain additional mutations that could mask or interfere with the effects of the global regulator mutations. The uniform genetic background of the Keio collection mutants enabled the characterization of the physiological consequences of altered carbon and redox fluxes caused by each global regulator deletion, eliminating possible strain-dependent results. As a proof of concept, Δcra and ΔarcA mutants were subjected to further manipulations to obtain large amounts of succinate and 1,3-PDO, demonstrating that the metabolic backgrounds of the mutants were suitable for the synthesis of bioproducts.

A Metabolic Widget Adjusts the Phosphoenolpyruvate-Dependent Fructose Influx in Pseudomonas putida.

Fructose uptake in the soil bacterium Pseudomonas putida occurs through a canonical phosphoenolpyruvate (PEP)-dependent sugar transport system (PTSFru). The logic of the genetic circuit that rules its functioning is puzzling: the transcription of the fruBKA operon, encoding all the components of PTSFru, can escape the repression exerted by the catabolite repressor/activator protein Cra solely in the presence of intracellular fructose-1-P, an agonist formed only when fructose has been already transported. To study this apparently incongruous regulatory architecture, the changes in the transcriptome brought about by a seamless Δcra deletion in P. putida strain KT2440 were inspected under different culture conditions. The few genes found to be upregulated in the cra mutant unexpectedly included PP_3443, encoding a bona fide glyceraldehyde-3-P dehydrogenase. An in silico model was developed to explore emergent properties that could result from such connections between sugar uptake with Cra and PEP. Simulation of fructose transport revealed that sugar uptake called for an extra supply of PEP (obtained through the activity of PP_3443) that was kept (i.e., memorized) even when the carbohydrate disappeared from the medium. This feature was traced to the action of two sequential inverters that connect the availability of exogenous fructose to intracellular PEP levels via Cra/PP_3443. The loss of such memory caused a much longer lag phase in cells shifted from one growth condition to another. The term "metabolic widget" is proposed to describe a merged biochemical and regulatory patch that tailors a given node of the cell molecular network to suit species-specific physiological needs. IMPORTANCE The regulatory nodes that govern metabolic traffic in bacteria often show connectivities that could be deemed unnecessarily complex at a first glance. Being a soil dweller and plant colonizer, Pseudomonas putida frequently encounters fructose in the niches that it inhabits. As is the case with many other sugars, fructose is internalized by a dedicated phosphoenolpyruvate (PEP)-dependent transport system (PTSFru), the expression of which is repressed by the fructose-1-P-responding Cra regulatory protein. However, Cra also controls a glyceraldehyde-3-P dehydrogenase that fosters accumulation of PEP (i.e., the metabolic fuel for PTSFru). A simple model representing this metabolic and regulatory device revealed that such an unexpected connectivity allows cells to shift smoothly between fructose-rich and fructose-poor conditions. Therefore, although the metabolic networks that handle sugar (i.e., fructose) consumption look very similar in most eubacteria, the way in which their components are intertwined endows given microorganisms with emergent properties for meeting species-specific and niche-specific needs.

Necesidades y sobrecarga en cuidadores primarios informales de pacientes con traumatismo craneoencefálico / Needs and overload on primary informal caregivers burden of patients with head injury

El objetivo de esta investigación fue conocer la relación entre la sobrecarga y las necesidades de los cuidadores primarios informales de pacientes que sufrieron traumatismo craneoencefálico. En una muestra no probabilística se aplicaron dos instrumentos a 64 cuidadores primarios que miden las necesidades (cuestionario de necesidades familiares) y sobrecarga (entrevista de sobrecarga del cuidador de Zarit), bajo un diseño transversal correlacional. Los resultados no mostraron diferencias significativas entre las categorías de las variables sociodemográficas, perosí presentaron relevancia práctica en las variables de sobrecarga, importancia de necesidades y satisfacción de necesidades. Estos resultados parecen sugerir la posibilidad de guiar estrategias de intervención hacia la satisfacción de necesidades de soporte emocional e instrumental con el objetivo de disminuir la sobrecarga en el cuidador primario informal y favorecer la calidad de vida; no obstante, es importante validar estos resultados en muestras de mayor tamaño que permitan generalizar los hallazgos.

The objective of this research was to determine the relationship between the overload and the needs of primary informal caregivers of patients suffering from a traumatic brain injury. In a non-probability sample, two instruments were applied to 64 primary caregivers measuring needs (family needs questionnaire) and overload (interview Zarit caregiver's burden), under a correlational cross-sectional design.The results showed no significant differences between categories of socio demographic variables, but had considerable practical relevance in relation to the variables of overload, importance of needs and satisfaction needs. These results seem to suggest the possibility of guiding intervention strategies towards meeting needs of emotional and instrumental support in order to reduce the burden on informal primary caregiver and promote the quality of life, however, it is important to validate these findings in larger samples that discard the possible influence of chance.

Ações de promoção da saúde para os idosos: análise da legislação proposta pela Agência Nacional de Saúde Suplementar / Health promotion actions for the elderly: analysis of legislation proposed by the National Health Agency Supplementary

Aumento significativo da população idosa gera aumento das doenças ligadas a esse ciclo e maiores gastos. No setor privadode saúde, trabalha-se visando ao assistencialismo com ações curativas distanciando da promoção à saúde. Esse trabalho tem por objetivo analisar as ações e os programas de promoção da saúde e prevenção de agravos,direcionadas aos idosos, propostas pelas operadoras de saúde no âmbito da legislação vigente da Agência Nacional de Saúde Suplementar. Trata-se de um trabalho de revisão bibliográfica com utilização de dados secundá¡rios. Conclui-se que a legislação estabelecida pela agência age de forma ineficiente na regulamentação dos planos de saúde. Destacam-se falhas nas ofertas dosprogramas de promoção da saúde, além do descumprimento dos planos em relação à legislação estabelecida .

Significant increase in the elderly population leads to increased diseases linked tothis cycle, and higher expenses. In the private health sector work is aimed at the welfare away with curative health promotion.This work aims to analyze the actions and programs of health promotion and diseaseprevention, directed at seniors, offered by health insurance companies under the current legislation of the National Health Agency. Itis a work of literature review using secondary data. We conclude that the law laid downby the agency, acting inefficiently in the regulation of health plans. Highlight gaps inthe offerings of programs to promote health,beyond the failure of plans in relation to theestablished laws .

Fructose 1-phosphate is the one and only physiological effector of the Cra (FruR) regulator of Pseudomonas putida.

Fructose-1-phosphate (F1P) is the preferred effector of the catabolite repressor/activator (Cra) protein of the soil bacterium Pseudomonas putida but its ability to bind other metabolic intermediates in vivo is unclear. The Cra protein of this microorganism (Cra(PP)) was submitted to mobility shift assays with target DNA sequences (the PfruB promoter) and candidate effectors fructose-1,6-bisphosphate (FBP), glucose 6-phosphate (G6P), and fructose-6-phosphate (F6P). 1 mM F1P was sufficient to release most of the Cra protein from its operators but more than 10 mM of FBP or G6P was required to free the same complex. However, isothermal titration microcalorimetry failed to expose any specific interaction between Cra(PP) and FBP or G6P. To solve this paradox, transcriptional activity of a PfruB-lacZ fusion was measured in wild-type and ΔfruB cells growing on substrates that change the intracellular concentrations of F1P and FBP. The data indicated that PfruB activity was stimulated by fructose but not by glucose or succinate. This suggested that Cra(PP) represses expression in vivo of the cognate fruBKA operon in a fashion dependent just on F1P, ruling out any other physiological effector. Molecular docking and dynamic simulations of the Cra-agonist interaction indicated that both metabolites can bind the repressor, but the breach in the relative affinity of Cra(PP) for F1P vs FBP is three orders of magnitude larger than the equivalent distance in the Escherichia coli protein. This assigns the Cra protein of P. putida the sole role of transducing the presence of fructose in the medium into a variety of direct and indirect physiological responses.