A multisite validation of a two hours antibiotic susceptibility flow cytometry assay directly from positive blood cultures.

BACKGROUND: Rapid antimicrobial susceptibility testing (AST) is urgently needed to provide safer treatment to counteract antimicrobial resistance. This is critical in septic patients, because resistance increases empiric therapy uncertainty and the risk of a poor outcome. We validate a novel 2h flow cytometry AST assay directly from positive blood cultures (PBC) by using a room temperature stable FASTgramneg and FASTgrampos kits (FASTinov® Porto, Portugal) in three sites: FASTinov (site-1), Hospital Ramon y Cajal, Madrid, Spain (site-2) and Centro Hospitalar S. João, Porto, Portugal (site-3). A total of 670 PBC were included: 333 spiked (site-1) and 337 clinical PBC (151 site-2 and 186 site-3): 367 gram-negative and 303 gram-positive. Manufacturer instructions were followed for sample preparation, panel inoculation, incubation (1h/37ºC) and flow cytometry analysis using CytoFlex (Site-1 and -2) or DxFlex (site-3) both instruments from Beckman-Coulter, USA. RESULTS: A proprietary software (bioFAST) was used to immediately generate a susceptibility report in less than 2 h. In parallel, samples were processed according to reference AST methods (disk diffusion and/or microdilution) and interpreted with EUCAST and CLSI criteria. Additionally, ten samples were spiked in all sites for inter-laboratory reproducibility. Sensitivity and specificity were >95% for all antimicrobials. Reproducibility was 96.8%/95.0% for FASTgramneg and 95.1%/95.1% for FASTgrampos regarding EUCAST/CLSI criteria, respectively. CONCLUSION: FASTinov® kits consistently provide ultra-rapid AST in 2h with high accuracy and reproducibility on both Gram-negative and Gram-positive bacteria. This technology creates a new paradigm in bacterial infection management and holds the potential to significantly impact septic patient outcomes and antimicrobial stewardship.

Hypersensitivity to Non-Steroidal Anti-Inflammatory Drugs on a pediatric Portuguese cohort.

Summary: Background. Non-steroidal anti-inflammatory drugs (NSAID)/analgesics (paracetamol) are among the most common causes of drug hypersensitivity reactions in children, with a reported prevalence of around 0.3% in the pediatric population. Paracetamol and ibuprofen are the most commonly reported culprits in the pediatric population. Our objective was to describe the allergy workup to NSAID/paracetamol of a pediatric population monitored in an allergy outpatient clinic. Methods. Retrospective observational study by consulting the medical records of patients evaluated in a pediatric outpatient clinic with history of NSAID/paracetamol, between January 2016 to August 2022. Results. A total of 43 patients have been evaluated for NSAID/paracetamol suspected allergy: 53.5% females, mean age of 9.8 ± 5.1 years, 47.7% atopic. The drugs reported as culprits were: ibuprofen (75.6%), paracetamol (17.8%), metamizole (4.4%) and naproxen (2.2%) and clinical manifestations were mainly urticaria/angioedema and maculopapular exanthema. Skin tests were performed in 7 patients: paracetamol (n = 5) and metamizole (n = 2), which were all negative. Fourty-six drug provocation tests were performed: 28 with the culprit drug and 18 with an alternative one; only 2 were positive (ibuprofen - culprit NSAID group): one immediate periorbital angioedema and one delayed lip edema with oropharyngeal tightness. Conclusions. The investigation of allergy to NSAID/paracetamol in children remains a challenge. In our population, ibuprofen was the most common NSAID reported. There were only 2 (4.3%) mild reactions on DPT. We could allow the use of the culprit NSAID/analgesic in 11 patients and an alternative one in 9 patients. This study highlights the importance of DPT in children for a correct diagnosis of NSAID hypersensitivity and selection of an alternative drug.

Variations in heterochromatin content reveal important polymorphisms for studies of genetic improvement in garlic (Allium sativum L) / Conteúdo de heterocromatina revela polimorfismos importantes para o melhoramento genético do alho (Allium sativum L)

Allium sativum L. is an herb of the Alliaceae family with a specific taste and aroma and medicinal and nutraceutical properties that are widely marketed in several countries. Brazil is one of the largest importers of garlic in the world, despite of its production is restricted and limited to internal consumption. Thus, explore the genetic diversity of commercial garlic conserved at germplasm banks is essential to generate additional genetic information about its economically important crop. A suitable tool for this purpose is the cytogenetic characterisation of these accessions. This study aimed to characterise the cytogenetic diversity among seven accessions of garlic from a Germplasm Bank in Brazil. The karyotypes were obtained by conventional staining and with chromomycin A3 (CMA) and 4,6-diamidino-2-phenylindole (DAPI) fluorochromes. All accessions analysed showed chromosome number 2n= 16, karyotype formula 6M+2SM, symmetrical karyotypes, reticulate interphase nuclei, and chromosomes with uniform chromatin condensation from prophase to metaphase. The fluorochromes staining showed differences in the amount and distribution of heterochromatin along the chromosomes and between accessions studied. Based on the distribution pattern of these small polymorphisms, it was possible to separate the seven accessions into three groups. It was also possible to differentiate some of the accessions individually. One of the results obtained showed a heteromorphic distension of the nucleolar organiser region observed on the chromosome pairs 6 or 7 with peculiar characteristics. It was suggested for example, that the heteromorphic block of heterochromatin (CMA+++/DAPI-) on chromosome 6 of the "Branco Mineiro Piauí" accession can be used as a marker to identify this genotype or may be associated with some character of economic interest.

Allium sativum L. é uma erva da família Alliaceae com sabor e aroma específicos e propriedades medicinais e nutracêuticas amplamente comercializada em diversos países. O Brasil é um dos maiores importadores de alho do mundo, apesar da sua produção ser restrita e limitada ao consumo interno. Assim, explorar a diversidade genética do alho comercial conservado em bancos de germoplasma é essencial para fornecer informações genéticas adicionais acerca dessa cultura economicamente importante. Uma ferramenta adequada para esse fim é a caracterização citogenética desses acessos. Este estudo teve como objetivo caracterizar a diversidade citogenética entre sete acessos de alho de um Banco de Germoplasma no Brasil. Os cariótipos foram obtidos por coloração convencional e com os fluorocromos de cromomicina A3 (CMA) e 4,6-diamidino-2-fenilindol (DAPI). Todos os acessos analisados apresentaram número cromossômico 2n = 16, fórmula cariotípica 6M + 2SM, cariótipos simétricos, núcleos reticulados em intérfase e cromossomos com condensação uniforme da cromatina da prófase para a metáfase. A coloração com fluorocromos mostrou diferenças na quantidade e distribuição de heterocromatina ao longo dos cromossomos e entre os acessos estudados. Com base no padrão de distribuição desses pequenos polimorfismos, foi possível separar os sete acessos em três grupos. Também foi possível diferenciar individualmente alguns dos acessos. Um dos resultados obtidos mostrou distensão heteromórfica da região organizadora nucleolar observada nos pares dos cromossomos 6 ou 7 com características peculiares. Foi sugerido, por exemplo, que o bloco heteromórfico de heterocromatina (CMA +++ / DAPI-) no cromossomo 6 do acesso “Branco Mineiro Piauí” pode ser usado como um marcador para identificar esse genótipo ou pode estar associado a algum caráter de interesse econômico.

Variations in heterochromatin content reveal important polymorphisms for studies of genetic improvement in garlic (Allium sativum L.)

Abstract Allium sativum L. is an herb of the Alliaceae family with a specific taste and aroma and medicinal and nutraceutical properties that are widely marketed in several countries. Brazil is one of the largest importers of garlic in the world, despite of its production is restricted and limited to internal consumption. Thus, explore the genetic diversity of commercial garlic conserved at germplasm banks is essential to generate additional genetic information about its economically important crop. A suitable tool for this purpose is the cytogenetic characterisation of these accessions. This study aimed to characterise the cytogenetic diversity among seven accessions of garlic from a Germplasm Bank in Brazil. The karyotypes were obtained by conventional staining and with chromomycin A3 (CMA) and 4,6-diamidino-2-phenylindole (DAPI) fluorochromes. All accessions analysed showed chromosome number 2n = 16, karyotype formula 6M+2SM, symmetrical karyotypes, reticulate interphase nuclei, and chromosomes with uniform chromatin condensation from prophase to metaphase. The fluorochromes staining showed differences in the amount and distribution of heterochromatin along the chromosomes and between accessions studied. Based on the distribution pattern of these small polymorphisms, it was possible to separate the seven accessions into three groups. It was also possible to differentiate some of the accessions individually. One of the results obtained showed a heteromorphic distension of the nucleolar organiser region observed on the chromosome pairs 6 or 7 with peculiar characteristics. It was suggested for example, that the heteromorphic block of heterochromatin (CMA+++/DAPI-) on chromosome 6 of the Branco Mineiro Piauí accession can be used as a marker to identify this genotype or may be associated with some character of economic interest.

Resumo Allium sativum L. é uma erva da família Alliaceae com sabor e aroma específicos e propriedades medicinais e nutracêuticas amplamente comercializada em diversos países. O Brasil é um dos maiores importadores de alho do mundo, apesar da sua produção ser restrita e limitada ao consumo interno. Assim, explorar a diversidade genética do alho comercial conservado em bancos de germoplasma é essencial para fornecer informações genéticas adicionais acerca dessa cultura economicamente importante. Uma ferramenta adequada para esse fim é a caracterização citogenética desses acessos. Este estudo teve como objetivo caracterizar a diversidade citogenética entre sete acessos de alho de um Banco de Germoplasma no Brasil. Os cariótipos foram obtidos por coloração convencional e com os fluorocromos de cromomicina A3 (CMA) e 4,6-diamidino-2-fenilindol (DAPI). Todos os acessos analisados apresentaram número cromossômico 2n = 16, fórmula cariotípica 6M + 2SM, cariótipos simétricos, núcleos reticulados em intérfase e cromossomos com condensação uniforme da cromatina da prófase para a metáfase. A coloração com fluorocromos mostrou diferenças na quantidade e distribuição de heterocromatina ao longo dos cromossomos e entre os acessos estudados. Com base no padrão de distribuição desses pequenos polimorfismos, foi possível separar os sete acessos em três grupos. Também foi possível diferenciar individualmente alguns dos acessos. Um dos resultados obtidos mostrou distensão heteromórfica da região organizadora nucleolar observada nos pares dos cromossomos 6 ou 7 com características peculiares. Foi sugerido, por exemplo, que o bloco heteromórfico de heterocromatina (CMA +++ / DAPI-) no cromossomo 6 do acesso Branco Mineiro Piauí pode ser usado como um marcador para identificar esse genótipo ou pode estar associado a algum caráter de interesse econômico.

Variations in heterochromatin content reveal important polymorphisms for studies of genetic improvement in garlic (Allium sativum L. ) / Conteúdo de heterocromatina revela polimorfismos importantes para o melhoramento genético do alho (Allium sativum L. )

Abstract Allium sativum L. is an herb of the Alliaceae family with a specific taste and aroma and medicinal and nutraceutical properties that are widely marketed in several countries. Brazil is one of the largest importers of garlic in the world, despite of its production is restricted and limited to internal consumption. Thus, explore the genetic diversity of commercial garlic conserved at germplasm banks is essential to generate additional genetic information about its economically important crop. A suitable tool for this purpose is the cytogenetic characterisation of these accessions. This study aimed to characterise the cytogenetic diversity among seven accessions of garlic from a Germplasm Bank in Brazil. The karyotypes were obtained by conventional staining and with chromomycin A3 (CMA) and 4,6-diamidino-2-phenylindole (DAPI) fluorochromes. All accessions analysed showed chromosome number 2n = 16, karyotype formula 6M+2SM, symmetrical karyotypes, reticulate interphase nuclei, and chromosomes with uniform chromatin condensation from prophase to metaphase. The fluorochromes staining showed differences in the amount and distribution of heterochromatin along the chromosomes and between accessions studied. Based on the distribution pattern of these small polymorphisms, it was possible to separate the seven accessions into three groups. It was also possible to differentiate some of the accessions individually. One of the results obtained showed a heteromorphic distension of the nucleolar organiser region observed on the chromosome pairs 6 or 7 with peculiar characteristics. It was suggested for example, that the heteromorphic block of heterochromatin (CMA+++/DAPI-) on chromosome 6 of the "Branco Mineiro Piauí" accession can be used as a marker to identify this genotype or may be associated with some character of economic interest.

Resumo Allium sativum L. é uma erva da família Alliaceae com sabor e aroma específicos e propriedades medicinais e nutracêuticas amplamente comercializada em diversos países. O Brasil é um dos maiores importadores de alho do mundo, apesar da sua produção ser restrita e limitada ao consumo interno. Assim, explorar a diversidade genética do alho comercial conservado em bancos de germoplasma é essencial para fornecer informações genéticas adicionais acerca dessa cultura economicamente importante. Uma ferramenta adequada para esse fim é a caracterização citogenética desses acessos. Este estudo teve como objetivo caracterizar a diversidade citogenética entre sete acessos de alho de um Banco de Germoplasma no Brasil. Os cariótipos foram obtidos por coloração convencional e com os fluorocromos de cromomicina A3 (CMA) e 4,6-diamidino-2-fenilindol (DAPI). Todos os acessos analisados ​​apresentaram número cromossômico 2n = 16, fórmula cariotípica 6M + 2SM, cariótipos simétricos, núcleos reticulados em intérfase e cromossomos com condensação uniforme da cromatina da prófase para a metáfase. A coloração com fluorocromos mostrou diferenças na quantidade e distribuição de heterocromatina ao longo dos cromossomos e entre os acessos estudados. Com base no padrão de distribuição desses pequenos polimorfismos, foi possível separar os sete acessos em três grupos. Também foi possível diferenciar individualmente alguns dos acessos. Um dos resultados obtidos mostrou distensão heteromórfica da região organizadora nucleolar observada nos pares dos cromossomos 6 ou 7 com características peculiares. Foi sugerido, por exemplo, que o bloco heteromórfico de heterocromatina (CMA +++ / DAPI-) no cromossomo 6 do acesso "Branco Mineiro Piauí" pode ser usado como um marcador para identificar esse genótipo ou pode estar associado a algum caráter de interesse econômico.

Behavioral effects of a low molecular weight peptide fraction from Phaseolus vulgaris in rats.

Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (<3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 µg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, α-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.

Marginal and Internal Misfit of Occlusal Veneers Made in Resin-matrix Ceramics.

OBJECTIVE: Considering that misfit is a significant predictor of the clinical success of indirect restorations, the objective of this study was to evaluate the marginal and internal misfit of two computer-aided design and manufacturing (CAD/CAM) RMC ceramic materials used as occlusal veneers (OVs) of different thicknesses. METHODS AND MATERIALS: A CAD model of a mandibular first molar was obtained and OV preparations 0.5-, 1.0-, and 1.5-mm thick were modeled and milled in two different materials (n=10/group): resin nanoceramic (RNC) and polymer-infiltrated ceramic network (PICN). Using the same CAD model, tooth preparations were milled in fiber-reinforced epoxy resin (n=20/thickness). The marginal and internal misfit of the restorations was assessed by X-ray microtomography. The measurements of the marginal gap (MG) and absolute marginal discrepancy were performed in two locations on each slice, whereas internal gap (IG) measurements were performed at ten locations on each slice. The data obtained were analyzed using two-way analysis of variance and Tukey post-hoc tests (α=0.05). RESULTS: No significant effect was attributable to the material type or material-thickness interaction for the MG, absolute marginal discrepancy (AMD), or IG (p>0.05). However, the thickness significantly affected the IG of the restorations (p<0.05). CAD/CAM RNC and PICN systems presented similar MG and AMD for OVs 0.5-, 1.0-, and 1.5-mm thick. However, the IG varied between thicknesses.

Impact of a large dam on reproduction of a non-migratory teleost species, Acestrorhynchus lacustris (Characiformes: Acestrorhynchidae) / Impacto de uma grande barragem na reprodução de uma espécie de teleósteo não migradora, Acestrorhynchus lacustris (Characiformes: Acestrorhynchidae)

The release of water from the reservoir hypolimnion, lower concentration of oxygen and the anthropogenic regulation of the river flow, could affect the reproduction of fish, especially migratory species. However, little is known about the effects of these changes in water on non-migratory species. In this sense, the reproduction of Acestrohynchus lacustris was evaluated in two sections of São Francisco River, Minas Gerais, Brazil. Section 1, located immediately downstream from Três Marias Dam (18°09'31.65"S and 45°13'36.00"W) and section 2, located at the confluence of the São Francisco and the Abaeté Rivers (18°02'47.78"S and 45°1057.95"W). For this, we obtained the physico-chemical parameters of water of each study section. Additionally, biometric data and biological indices ofall specimens were measured. Fecundity and follicles diameters were measured in females. Temperature, dissolvedoxygen and flow showed lower values in section 1. Fish captured in this section, had lower values of GSI in bothsexes, and females presented decreased values of fecundity and follicles diameter. This species showed reproductiveactivity in the two sections analyzed, however, in section 1 where the temperature and dissolved oxygen presentedsignificant lower values, the reproductive capacity of A. lacustris, was negatively affected.

A liberação de água do hipolímnio do reservatório, baixa concentração de oxigênio e a regulação antropogênica do fluxo do rio podem afetar a reprodução de peixes, principalmente espécies migradoras. No entanto, pouco se sabe sobre os efeitos dessas mudanças nas condições da água em espécies não migradoras. A reprodução de Acestrohynchus lacustris foi avaliada em duas seções do rio São Francisco. Seção 1, localizada imediatamente a jusante da barragem de Três Marias e seção 2, localizada na confluência dos rios São Francisco e Abaeté. Para isso, foram obtidos os parâmetros físico-químicos da água de cada seção do estudo. Além disso, dados biométricos e índices biológicos de todos os peixes capturados foram obtidos. Adicionalmente, nas fêmeas foram medidos os diâmetros dos folículos vitelogênicos e a fecundidade. Temperatura, oxigênio dissolvido e fluxo apresentaram valores mais baixos na seção 1. Os peixes capturados nesta seção apresentaram menores valores de IGS em ambos os sexos, e as fêmeas apresentaram menores valores de fecundidade e diâmetro dos folículos. Essa espécie apresentou atividade reprodutiva nas duas seções analisadas, porém, na seção 1, onde os parâmetros da água apresentam piores condições para o processo reprodutivo de peixes, a capacidade reprodutiva de A. lacustris foi afetada negativamente.

Impact of a large dam on reproduction of a non-migratory teleost species, Acestrorhynchus lacustris (Characiformes: Acestrorhynchidae)

Abstract The release of water from the reservoir hypolimnion, lower concentration of oxygen and the anthropogenic regulation of the river flow, could affect the reproduction of fish, especially migratory species. However, little is known about the effects of these changes in water on non-migratory species. In this sense, the reproduction of Acestrohynchus lacustris was evaluated in two sections of São Francisco River, Minas Gerais, Brazil. Section 1, located immediately downstream from Três Marias Dam (18°0931.65S and 45°1336.00W) and section 2, located at the confluence of the São Francisco and the Abaeté Rivers (18°0247.78S and 45°1057.95W). For this, we obtained the physico-chemical parameters of water of each study section. Additionally, biometric data and biological indices of all specimens were measured. Fecundity and follicles diameters were measured in females. Temperature, dissolved oxygen and flow showed lower values in section 1. Fish captured in this section, had lower values of GSI in both sexes, and females presented decreased values of fecundity and follicles diameter. This species showed reproductive activity in the two sections analyzed, however, in section 1 where the temperature and dissolved oxygen presented significant lower values, the reproductive capacity of A. lacustris, was negatively affected.

Resumo A liberação de água do hipolímnio do reservatório, baixa concentração de oxigênio e a regulação antropogênica do fluxo do rio podem afetar a reprodução de peixes, principalmente espécies migradoras. No entanto, pouco se sabe sobre os efeitos dessas mudanças nas condições da água em espécies não migradoras. A reprodução de Acestrohynchus lacustris foi avaliada em duas seções do rio São Francisco. Seção 1, localizada imediatamente a jusante da barragem de Três Marias e seção 2, localizada na confluência dos rios São Francisco e Abaeté. Para isso, foram obtidos os parâmetros físico-químicos da água de cada seção do estudo. Além disso, dados biométricos e índices biológicos de todos os peixes capturados foram obtidos. Adicionalmente, nas fêmeas foram medidos os diâmetros dos folículos vitelogênicos e a fecundidade. Temperatura, oxigênio dissolvido e fluxo apresentaram valores mais baixos na seção 1. Os peixes capturados nesta seção apresentaram menores valores de IGS em ambos os sexos, e as fêmeas apresentaram menores valores de fecundidade e diâmetro dos folículos. Essa espécie apresentou atividade reprodutiva nas duas seções analisadas, porém, na seção 1, onde os parâmetros da água apresentam piores condições para o processo reprodutivo de peixes, a capacidade reprodutiva de A. lacustris foi afetada negativamente.

Impact of a large dam on reproduction of a non-migratory teleost species, Acestrorhynchus lacustris (Characiformes: Acestrorhynchidae) / Impacto de uma grande barragem na reprodução de uma espécie de teleósteo não migradora, Acestrorhynchus lacustris (Characiformes: Acestrorhynchidae)

The release of water from the reservoir hypolimnion, lower concentration of oxygen and the anthropogenic regulation of the river flow, could affect the reproduction of fish, especially migratory species. However, little is known about the effects of these changes in water on non-migratory species. In this sense, the reproduction of Acestrohynchus lacustris was evaluated in two sections of São Francisco River, Minas Gerais, Brazil. Section 1, located immediately downstream from Três Marias Dam (18°09'31.65"S and 45°13'36.00"W) and section 2, located at the confluence of the São Francisco and the Abaeté Rivers (18°02'47.78"S and 45°10'57.95"W). For this, we obtained the physico-chemical parameters of water of each study section. Additionally, biometric data and biological indices of all specimens were measured. Fecundity and follicles diameters were measured in females. Temperature, dissolved oxygen and flow showed lower values in section 1. Fish captured in this section, had lower values of GSI in both sexes, and females presented decreased values of fecundity and follicles diameter. This species showed reproductive activity in the two sections analyzed, however, in section 1 where the temperature and dissolved oxygen presented significant lower values, the reproductive capacity of A. lacustris, was negatively affected.

A liberação de água do hipolímnio do reservatório, baixa concentração de oxigênio e a regulação antropogênica do fluxo do rio podem afetar a reprodução de peixes, principalmente espécies migradoras. No entanto, pouco se sabe sobre os efeitos dessas mudanças nas condições da água em espécies não migradoras. A reprodução de Acestrohynchus lacustris foi avaliada em duas seções do rio São Francisco. Seção 1, localizada imediatamente a jusante da barragem de Três Marias e seção 2, localizada na confluência dos rios São Francisco e Abaeté. Para isso, foram obtidos os parâmetros físico-químicos da água de cada seção do estudo. Além disso, dados biométricos e índices biológicos de todos os peixes capturados foram obtidos. Adicionalmente, nas fêmeas foram medidos os diâmetros dos folículos vitelogênicos e a fecundidade. Temperatura, oxigênio dissolvido e fluxo apresentaram valores mais baixos na seção 1. Os peixes capturados nesta seção apresentaram menores valores de IGS em ambos os sexos, e as fêmeas apresentaram menores valores de fecundidade e diâmetro dos folículos. Essa espécie apresentou atividade reprodutiva nas duas seções analisadas, porém, na seção 1, onde os parâmetros da água apresentam piores condições para o processo reprodutivo de peixes, a capacidade reprodutiva de A. lacustris foi afetada negativamente.

Behavioral effects of a low molecular weight peptide fraction from Phaseolus vulgaris in rats

Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (<3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 µg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, α-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.

Impact of a large dam on reproduction of a non-migratory teleost species, Acestrorhynchus lacustris (Characiformes: Acestrorhynchidae).

The release of water from the reservoir hypolimnion, lower concentration of oxygen and the anthropogenic regulation of the river flow, could affect the reproduction of fish, especially migratory species. However, little is known about the effects of these changes in water on non-migratory species. In this sense, the reproduction of Acestrohynchus lacustris was evaluated in two sections of São Francisco River, Minas Gerais, Brazil. Section 1, located immediately downstream from Três Marias Dam (18°09'31.65"S and 45°13'36.00"W) and section 2, located at the confluence of the São Francisco and the Abaeté Rivers (18°02'47.78"S and 45°10'57.95"W). For this, we obtained the physico-chemical parameters of water of each study section. Additionally, biometric data and biological indices of all specimens were measured. Fecundity and follicles diameters were measured in females. Temperature, dissolved oxygen and flow showed lower values in section 1. Fish captured in this section, had lower values of GSI in both sexes, and females presented decreased values of fecundity and follicles diameter. This species showed reproductive activity in the two sections analyzed, however, in section 1 where the temperature and dissolved oxygen presented significant lower values, the reproductive capacity of A. lacustris, was negatively affected.

Variations in heterochromatin content reveal important polymorphisms for studies of genetic improvement in garlic (Allium sativum L.).

Allium sativum L. is an herb of the Alliaceae family with a specific taste and aroma and medicinal and nutraceutical properties that are widely marketed in several countries. Brazil is one of the largest importers of garlic in the world, despite of its production is restricted and limited to internal consumption. Thus, explore the genetic diversity of commercial garlic conserved at germplasm banks is essential to generate additional genetic information about its economically important crop. A suitable tool for this purpose is the cytogenetic characterisation of these accessions. This study aimed to characterise the cytogenetic diversity among seven accessions of garlic from a Germplasm Bank in Brazil. The karyotypes were obtained by conventional staining and with chromomycin A3 (CMA) and 4,6-diamidino-2-phenylindole (DAPI) fluorochromes. All accessions analysed showed chromosome number 2n = 16, karyotype formula 6M+2SM, symmetrical karyotypes, reticulate interphase nuclei, and chromosomes with uniform chromatin condensation from prophase to metaphase. The fluorochromes staining showed differences in the amount and distribution of heterochromatin along the chromosomes and between accessions studied. Based on the distribution pattern of these small polymorphisms, it was possible to separate the seven accessions into three groups. It was also possible to differentiate some of the accessions individually. One of the results obtained showed a heteromorphic distension of the nucleolar organiser region observed on the chromosome pairs 6 or 7 with peculiar characteristics. It was suggested for example, that the heteromorphic block of heterochromatin (CMA+++/DAPI-) on chromosome 6 of the "Branco Mineiro Piauí" accession can be used as a marker to identify this genotype or may be associated with some character of economic interest.

Oxidonitrergic and antioxidant effects of a low molecular weight peptide fraction from hardened bean (Phaseolus vulgaris) on endothelium.

About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction <3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 µg/mL PV3. Oxidative stress was provoked by 3% H2O2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO- and endothelium-dependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 µg/mL PV3. PV3 at 30 µg/mL increased cell viability, while cytotoxicity was observed only with 250 µg/mL PV3. PV3 at 10 µg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H2O2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release.

Water requirements of beef production can be reduced by genetic selection.

Growing concerns regarding sustainability in agriculture include the availability of drinking water, which is putting pressure on livestock production, especially the beef sector, for more efficient practices. Thus, genetic parameters were estimated for traits related to water intake and water use efficiency in Senepol cattle. Senepol females (n = 925) and males (n = 191) were evaluated in performance tests carried out from 2014 to 2019. Daily dry matter intake (DMI) and water intake (WI) were recorded by electronic feed and water bunks (Intergado Ltd.). Other traits assessed included average daily gain (ADG); mid-test metabolic BW (BW0.75); residual water intake based on ADG (RWIADG), estimated as the residual of the linear regression equation of WI on ADG and BW0.75; residual water intake based on DMI (RWIDMI), estimated as the residual of the linear regression equation of WI on DMI and BW0.75 (RWIDMI); water conversion ratio (= WI/ADG); gross water efficiency (GWE = ADG/WI); residual feed intake estimated as the residual of the linear regression equation of DMI on ADG and BW0.75 (RFI); feed conversion ratio (= DMI/ADG) and gross feed efficiency. Genetic (co)variances were estimated with bivariate analyses. The heritabilities for WI, RWIADG and RWIDMI were 0.38, 0.36 and 0.33, respectively. Water conversion ratio, RWIADG and RWIDMI showed positive genetic and phenotypic correlations with WI, whereas GWE was negatively correlated with WI, suggesting that traits related to water use efficiency may be useful to identify cattle with reduced WI. Water intake showed positive genetic (r = 0.79) and phenotypic (r = 0.60) correlations with DMI, suggesting the use of WI to estimate DMI in future studies. Both RWIADG and RWIDMI were genetically correlated with RFI (0.67 and 0.57, respectively) and ADG (0.49 and 0.44, respectively), showing that RWI is positively associated with feed efficiency, but has an antagonistic relationship with growth. This antagonism, however, may be managed using selection indexes. Genetic improvement of water use efficiency in Senepol cattle is possible through selection and may reduce the water requirements of beef production systems.

Genetic parameters for performance, feed efficiency, and carcass traits in Senepol heifers.

Improving feed efficiency is a key breeding goal in the beef cattle industry. In this study, we estimated the genetic parameters for feed efficiency and carcass traits in Senepol cattle raised in tropical regions. Various indicators of feed efficiency [gain to feed ratio (G:F), feed conversion ratio (FCR), residual weight gain (RG), residual intake and body weight gain (RIG), and residual feed intake (RFI)] as well as growth [final BW, average daily gain (ADG), and DM intake (DMI)], and carcass [rib-eye area (REA), backfat thickness (BF), intramuscular fat score, and carcass conformation score] traits were included in the study. After data editing, records from 1 393 heifers obtained between 2009 and 2018 were used for the analyses. We fitted an animal model that included contemporary group (animals from the same farm that were evaluated in the same test season) as the fixed effect, and a linear effect of animal age at the beginning of the test as a covariate; in addition to random direct additive genetic and residual effects. The (co)variance components were estimated by Bayesian inference in uni- and bivariate analyses. Our results showed that feed efficiency indicators derived from residual variables such as RG, RIG, and RFI can be improved through genetic selection (h2 = 0.14 ± 0.06, 0.13 ± 0.06, and 0.20 ± 0.08, respectively). Variables calculated as ratios such as G:F and FCR were more influenced by environmental factors (h2 = 0.08 ± 0.05 and 0.09 ± 0.05), and were, therefore, less suitable for use in breeding programs. The traits with the greatest and impact on genetic progress in feed efficiency were ADG, REA, and BF. The traits with the greatest and least impact on growth and carcass traits were RG and RFI, respectively. Selection for feed efficiency will result in distinct overall effects on the growth and carcass traits of Senepol heifers. Direct selection for lower RFI may reduce DMI and increase carcass fatness at the finishing stage, but it might also result in reduced growth and muscle deposition. Residual BW gain is associated with the highest weight gain and zero impact on REA and BF, however, it is linked to higher feed consumption. Thus, the most suitable feed efficiency indicator was RIG, as it promoted the greatest decrease in feed intake concomitant with faster growth, with a similar impact on carcass traits when compared to the other feed efficiency indicators.