The latest freshwater giants: a new Peltocephalus (Pleurodira: Podocnemididae) turtle from the Late Pleistocene of the Brazilian Amazon.

Overkill of large mammals is recognized as a key driver of Pleistocene megafaunal extinctions in the Americas and Australia. While this phenomenon primarily affected mega-mammals, its impact on large Quaternary reptiles has been debated. Freshwater turtles, due to the scarcity of giant forms in the Quaternary record, have been largely neglected in such discussions. Here we present a new giant podocnemidid turtle, Peltocephalus maturin sp. nov., from the Late Pleistocene Rio Madeira Formation in the Brazilian Amazon, that challenges this assumption. Morphological and phylogenetic analyses of the holotype, a massive partial lower jaw, reveal close affinities to extant Amazonian species and suggest an omnivorous diet. Body size regressions indicate Pe. maturin possibly reached about 180 cm in carapace length and is among the largest freshwater turtles ever found. This finding presents the latest known occurrence of giant freshwater turtles, hinting at coexistence with early human inhabitants in the Amazon.

Droplet-based logic gates simulation of viscoelastic fluids under electric field.

Nano and microfluidic technologies have shown great promise in the development of controlled drug delivery systems and the creation of microfluidic devices with logic-like functionalities. Here, we focused on investigating a droplet-based logic gate that can be used for automating medical diagnostic assays. This logic gate uses viscoelastic fluids, which are particularly relevant since bio-fluids exhibit viscoelastic properties. The operation of the logic gate is determined by evaluating various parameters, including the Weissenberg number, the Capillary number, and geometric factors. To effectively classify the logic gates operational conditions, we employed a deep learning classification to develop a reduced-order model. This approach accelerates the prediction of operating conditions, eliminating the need for complex simulations. Moreover, the deep learning model allows for the combination of different AND/OR branches, further enhancing the versatility of the logic gate. We also found that non-operating regions, where the logic gate does not function properly, can be transformed into operational regions by applying an external force. By utilizing an electrical induction technique, we demonstrated that the application of an electric field can repel or attract droplets, thereby improving the performance of the logic gate. Overall, our research shows the potential of the droplet-based logic gates in the field of medical diagnostics. The integration of deep learning classification algorithms enables rapid evaluation of operational conditions and facilitates the design of complex logic circuits. Additionally, the introduction of external forces and electrical induction techniques opens up new possibilities for enhancing the functionality and reliability of these logic gates.

Oxidative and osmotolerant effects in Salvator merianae (Squamata: Teiidae) red blood cells during hibernation

Abstract Hibernation is a natural condition of animals that lives in the temperate zone, although some tropical lizards also experience hibernation annually, such as the lizard native from South America, Salvator merianae, or tegu lizard. Even though physiological and metabolic characteristic associated with hibernation have been extensively studied, possible alterations in the red blood cells (RBC) integrity during this period remains unclear. Dehydration and fasting are natural consequences of hibernating for several months and it could be related to some cellular modifications. In this study, we investigated if the osmotic tolerance of RBCs of tegu lizard under hibernation is different from the cells obtained from animals while normal activity. Additionally, we indirectly investigated if the RBCs membrane of hibernating tegus could be associated with oxidation by quantifying oxidized biomolecules and the activity of antioxidant enzymes. Our findings suggest that RBCs are more fragile during the hibernation period, although we did not find evidence of an oxidative stress scenario associated with the accentuated fragility. Even though we did not exclude the possibility of oxidative damage during hibernation, we suggested that an increased RBCs volume as a consequence of hypoosmotic blood during hibernation could also affect RBCs integrity as noted.

Resumo A hibernação é uma condição natural dos animais que vivem na zona temperada, embora alguns lagartos tropicais também experenciem hibernação anualmente, como é o caso do lagarto nativo da América do Sul, Salvator merianae ou teiú. Embora as características fisiológicas e metabólicas associadas à hibernação tenham sido amplamente estudadas, possíveis alterações na integridade das hemácias durante esse período ainda permanecem obscuras. A desidratação e o jejum são consequências naturais da hibernação por vários meses e podem estar relacionadas a algumas modificações celulares. Neste estudo, investigamos se a tolerância osmótica de hemácias do lagarto teiú sob hibernação são diferentes das células obtidas de animais em atividade normal. Além disso, investigamos indiretamente por meio da quantificação de biomoléculas oxidadas e da atividade de enzimas antioxidantes se a membrana das hemácias dos teiús em hibernação poderia estar associada à oxidação. Nossos resultados sugerem que as hemácias possuem maior fragilidade durante o período de hibernação, embora não tenhamos encontrado evidências de um cenário de estresse oxidativo associado à essa fragilidade acentuada. Embora não tenhamos excluído a possibilidade de dano oxidativo durante a hibernação, sugerimos que um aumento no volume das hemácias como consequência de sangue hipoosmótico durante a hibernação também poderia afetar a integridade de hemácias, tal como foi observado.

Oxidative and osmotolerant effects in Salvator merianae (Squamata: Teiidae) red blood cells during hibernation / Efeitos de oxidação e osmotolerância dos eritrócitos de Salvator merianae (Squamata: Teiidae) durante a hibernação

Abstract Hibernation is a natural condition of animals that lives in the temperate zone, although some tropical lizards also experience hibernation annually, such as the lizard native from South America, Salvator merianae, or "tegu" lizard. Even though physiological and metabolic characteristic associated with hibernation have been extensively studied, possible alterations in the red blood cells (RBC) integrity during this period remains unclear. Dehydration and fasting are natural consequences of hibernating for several months and it could be related to some cellular modifications. In this study, we investigated if the osmotic tolerance of RBCs of tegu lizard under hibernation is different from the cells obtained from animals while normal activity. Additionally, we indirectly investigated if the RBCs membrane of hibernating tegus could be associated with oxidation by quantifying oxidized biomolecules and the activity of antioxidant enzymes. Our findings suggest that RBCs are more fragile during the hibernation period, although we did not find evidence of an oxidative stress scenario associated with the accentuated fragility. Even though we did not exclude the possibility of oxidative damage during hibernation, we suggested that an increased RBCs volume as a consequence of hypoosmotic blood during hibernation could also affect RBCs integrity as noted.

Resumo A hibernação é uma condição natural dos animais que vivem na zona temperada, embora alguns lagartos tropicais também experenciem hibernação anualmente, como é o caso do lagarto nativo da América do Sul, Salvator merianae ou "teiú". Embora as características fisiológicas e metabólicas associadas à hibernação tenham sido amplamente estudadas, possíveis alterações na integridade das hemácias durante esse período ainda permanecem obscuras. A desidratação e o jejum são consequências naturais da hibernação por vários meses e podem estar relacionadas a algumas modificações celulares. Neste estudo, investigamos se a tolerância osmótica de hemácias do lagarto teiú sob hibernação são diferentes das células obtidas de animais em atividade normal. Além disso, investigamos indiretamente por meio da quantificação de biomoléculas oxidadas e da atividade de enzimas antioxidantes se a membrana das hemácias dos teiús em hibernação poderia estar associada à oxidação. Nossos resultados sugerem que as hemácias possuem maior fragilidade durante o período de hibernação, embora não tenhamos encontrado evidências de um cenário de estresse oxidativo associado à essa fragilidade acentuada. Embora não tenhamos excluído a possibilidade de dano oxidativo durante a hibernação, sugerimos que um aumento no volume das hemácias como consequência de sangue hipoosmótico durante a hibernação também poderia afetar a integridade de hemácias, tal como foi observado.

Is leaf fluctuating asymmetry related to plant and leaf size in Miconia albicans, a common Melastomataceae species? / A assimetria flutuante de folhas está relacionada ao tamanho da planta e da folha em um arbusto de savana neotropical?

Fluctuating asymmetry, defined as random differences between the two sides of a symmetrical structure, has been often related to development stress in both plants and animals. In plants, leaf fluctuating asymmetry has been related to stresses such as pollution and fire and may also be related to leaf growth and herbivory rates. We assessed whether leaf fluctuating asymmetry is related to plant and leaf size in Miconia albicans (Sw.) Triana (Melastomataceae), a common multi-stem Neotropical shrub, in a Brazilian savanna area. We collected 15 leaves from each of 70 individuals, and measured fluctuating asymmetry as the difference in area between the right and left sides of the leaves using the central vein as reference. To avoid spurious results due to measurement error, the division along the central vein was performed independently by three researchers. We also measured the basal area and height of each stem of the plant individuals. We used linear models to assess the relations between leaf fluctuating asymmetry, plant size and leaf size. No consistent relations were observed between leaf fluctuating asymmetry and plant size, as the analyses performed on the fluctuating asymmetry values obtained by the different researchers showed different results. However, relative fluctuating asymmetry values, obtained by dividing the fluctuating asymmetry by the total leaf area, tended to be smaller in larger leaves. It thus appears that, in the study species, fluctuating asymmetry is related to the developmental conditions faced by the individual leaves and not by the plant as a whole.

A assimetria flutuante, definida como diferenças aleatórias entre os dois lados de uma estrutura simétrica, tem sido frequentemente relacionada ao estresse de desenvolvimento em plantas e animais. Nas plantas, a assimetria flutuante foliar tem sido relacionada a estresses como poluição e fogo e também pode estar relacionada ao crescimento foliar e taxas de herbivoria. Nós avaliamos se a assimetria flutuante foliar está relacionada ao tamanho da planta e da folha em Miconia albicans (Sw.) Triana (Melastomataceae), um arbusto neotropical multicaule comum, em uma área de Cerrado. Coletamos 15 folhas de cada um dos 70 indivíduos e medimos a assimetria flutuante como a diferença de área entre os lados direito e esquerdo das folhas usando a nervura central como referência. Para evitar resultados espúrios devido ao erro de medição, a separação ao longo da nervura centra foi feita independentemente por três pesquisadoras/es. Também medimos a área basal e a altura de cada caule de cada planta. Usamos modelos lineares para avaliar as relações entre assimetria flutuante foliar, tamanho da planta e tamanho da folha. Não foram observadas relações consistentes entre a assimetria flutuante da folha e o tamanho da planta, pois as análises realizadas nos valores da assimetria flutuante obtidos pelas/os diferentes pesquisadoras/es mostraram resultados diferentes. No entanto, os valores de assimetria flutuante relativa, obtidos pela divisão da assimetria flutuante pela área foliar total, tenderam a ser menores nas folhas maiores. Assim, verifica-se que, na espécie estudada, a assimetria flutuante está relacionada às condições de desenvolvimento enfrentadas pelas folhas individuais e não pela planta como um todo.

Identification of APOB variants in samples from the brazilian population with hypercholesterolemic phenotype

BACKGROUND AND AIMS: Apolipoprotein B plays a crucial role in regulating plasma cholesterol by mediating the interaction of low-density lipoprotein (LDL) with LDL receptors in the liver. Inherited mutations in this gene may increase the risk of developing premature atherosclerotic cardiovascular disease, especially in individuals with familial hypercholesterolemia type 2 (FH2). The aim of this study is to identify APOB variants that may indicate pathogenicity in a sample of the Brazilian population using a data bank exome sequencing study by NGS in a Brazilian population phenotypically diagnosed by clinical and laboratory profile. This finding is going to improve genetic hypercholesteremia diagnosis. METHODS: High-quality DNA samples (n»300) were sequenced using an exon-targeted gene sequencing (ETGS) strategy to identify variants in FHrelated genes. Pathogenicity classification was based on criteria established by the American College of Medical Genetics and Genomics (ACMG), also using information from ClinVar and pathogenicity scores from previous association studies. RESULTS: A total of 121 variants were identified in APOB, of which four are novel variants missense (p.Thr626Asn, p.Ile2750Thr, p.Gln2078Lys and p.Met4184Arg). After curating pathogenicity scores, variants were classified according to the ACMG criteria. Among them four as pathogenic or likely pathogenic (p.Pro2739Leu, p.His1923Arg, p.Pro994Leu and p.Pro877Leu), and 21 variants had uncertain significance. Additionally, 92 previously known variants with uncertain significance were classified as benign or likely benign. The results were submitted to Clinvar for actualization of pathogenicity. CONCLUSIONS: These results improve the molecular diagnosis associating APOB variants with the clinical phenotype of hypercholesterolemia.

Oral amyloidosis: an update.

BACKGROUND: Amyloidosis is a disease characterized by the progressive deposition of abnormal proteins that can occur in any organ. In the oral cavity, the tongue is the most common affected site, usually causing macroglossia. Biopsy is essential for the diagnosis and the occurrence of its systemic form is mandatory to be investigated. This systematic review evaluated the existing information in the literature on Amyloidosis in the oral cavity to allow a more comprehensive and updated analysis of its clinicopathological characteristics, as well as to explore the main forms of treatment and prognostic factors. MATERIAL AND METHODS: Electronic searches were undertaken in five databases supplemented by manual scrutiny. RESULTS: A total of 111 studies were included with 158 individuals. CONCLUSIONS: The disease had a higher prevalence in women, the tongue was the most affected site, as well as the systemic form of the disease. The worst prognosis was for cases of systemic amyloidosis associated with multiple myeloma.

Genetic parameters for frame size and carcass traits in Nellore cattle.

Our study aimed to evaluate the frame size as a genetic selection criterion as well as the association of scores with carcass characteristics measured via ultrasound to the genetic improvement of carcass quality in Nellore cattle. The variance components were estimated using the restricted maximum likelihood method for the thoracic perimeter (TP), frame size score (FRAME), yearling weight (YW), visual scores of body structure (BS), muscularity (MUSC), and precocity (PREC). The carcass traits included loin eye area (REA), backfat thickness (BF), rump fat (FR), and intramuscular fat (IF). The estimated heritabilities ranged from moderate to high. The FRAME showed a positive genetic correlation with YW (0.82 ± 0.07), TP (0.58 ± 0.09), and BS score (0.62 ± 0.07), but it had a negative correlation with the MUSC and PREC visual scores (- 0.34 ± 0.09 and - 0.50 ± 0.08, respectively). The genetic correlation between FRAME and REA was positive (0.26 ± 0.09). However, FRAME presented a correlation close to zero with BF, RF, and IF. TP had a positive genetic correlation with body size traits such as YW (0.76 ± 0.05) and BS score (0.55 ± 0.08) and with muscularity traits such as REA (0.44 ± 0.01). Frame size has the potential to be used as a parameter in the selection of precocious beef cattle, but a study of its use is necessary to determine the ideal biotype for each production system.

Is leaf fluctuating asymmetry related to plant and leaf size in Miconia albicans, a common Melastomataceae species?

Fluctuating asymmetry, defined as random differences between the two sides of a symmetrical structure, has been often related to development stress in both plants and animals. In plants, leaf fluctuating asymmetry has been related to stresses such as pollution and fire and may also be related to leaf growth and herbivory rates. We assessed whether leaf fluctuating asymmetry is related to plant and leaf size in Miconia albicans (Sw.) Triana (Melastomataceae), a common multi-stem Neotropical shrub, in a Brazilian savanna area. We collected 15 leaves from each of 70 individuals, and measured fluctuating asymmetry as the difference in area between the right and left sides of the leaves using the central vein as reference. To avoid spurious results due to measurement error, the division along the central vein was performed independently by three researchers. We also measured the basal area and height of each stem of the plant individuals. We used linear models to assess the relations between leaf fluctuating asymmetry, plant size and leaf size. No consistent relations were observed between leaf fluctuating asymmetry and plant size, as the analyses performed on the fluctuating asymmetry values obtained by the different researchers showed different results. However, relative fluctuating asymmetry values, obtained by dividing the fluctuating asymmetry by the total leaf area, tended to be smaller in larger leaves. It thus appears that, in the study species, fluctuating asymmetry is related to the developmental conditions faced by the individual leaves and not by the plant as a whole.

Intestinal morphometry and performance of broiler chickens subjected to diets with buriti oil / [Morfometria intestinal e desempenho de frangos de corte submetidos a dietas com óleo de buriti]

This study aimed to evaluate the productive performance, carcass yield, and intestinal morphometry of broiler chickens subjected to diets with the inclusion of buriti oil. Buriti oil is an energy food that can be used in chicken feed to replace soybean oil, which has a high cost in production, also has its anti-inflammatory, antioxidant, and antimicrobial properties, which can provide benefits in chicken feed. 180 male broilers of the Ross lineage were used, distributed in a completely randomized design with 3 treatments and 6 replicates of 10 birds per experimental unit. The experimental phase took place from 14 to 28 days and levels 0; 0.75 and 1.50% of buriti oil were added to the corn and soybean-based feed. It can be inferred that the buriti oil-based diets in the diet with insertion of levels of 0%, 0.75% and 1.50% did not differ significantly for the productive performance variables, but in relation to the productive yield there was a significant difference for carcass yield accompanied by greater disposition of abdominal fat, thigh and heart fat, a common effect in the use of oils. Although no statistical differences were observed for most of the variables in the assessment of intestinal morphometry, there was an increase in the crypt depth of the duodenum and ileum and an increase in the muscle layer of the duodenum and jejunum, which may have been caused by some injury to the intestine and not by direct effect of buriti oil. In the other variables there was no difference between treatments and it is concluded that buriti oil did not harm the performance, yield and intestinal morphometry of broiler chickens from 14 to 28 days of age.

Objetivou-se, com essa pesquisa, avaliar o desempenho produtivo, o rendimento de carcaça e a morfometria intestinal de frangos de corte com dietas submetidas à inclusão do óleo de buriti. Trata-se de um alimento energético que pode ser utilizado na alimentação de frangos visando substituir o óleo de soja, o qual apresenta alto custo na produção. O óleo de buriti ainda possui suas propriedades anti-inflamatórias, antioxidantes, antimicrobianas, podendo conferir benefícios na alimentação de frangos. Foram utilizados 180 frangos de corte machos, da linhagem Ross, distribuídos em um delineamento inteiramente ao acaso, com três tratamentos e seis repetições de 10 aves por unidade experimental. A fase experimental ocorreu de 14 a 28 dias, e os níveis 0%, 0,75% e 1,50% de óleo de buriti foram inseridos na ração à base de milho e soja. Pode-se inferir que as dietas à base de óleo de buriti na dieta com inserção dos níveis de 0%, 0,75% e 1,50% não diferiram significativamente para as variáveis de desempenho produtivo, mas, em relação ao rendimento produtivo, houve uma diferença significativa para o rendimento de carcaça, acompanhado de maior disposição de gordura abdominal, gordura de sobrecoxa e coração, efeito comum na utilização de óleos. Apesar de não serem observadas diferenças estatísticas para a maioria das variáveis na avaliação da morfometria intestinal, houve aumento na profundidade de cripta do duodeno e do íleo, bem como aumento da camada muscular do duodeno e do jejuno, que podem ter sido provocados por alguma injúria no intestino e não por efeito direto do óleo de buriti. Nas demais variáveis, não houve diferença entre os tratamentos, e conclui-se que o óleo de buriti não causou prejuízo ao desempenho, ao rendimento e à morfometria intestinal de frangos de corte no período de 14 a 28 dias de idade.

Engineering topological phases in triple HgTe/CdTe quantum wells.

Quantum wells formed by layers of HgTe between Hg[Formula: see text]Cd[Formula: see text]Te barriers lead to two-dimensional (2D) topological insulators, as predicted by the BHZ model. Here, we theoretically and experimentally investigate the characteristics of triple HgTe quantum wells. We describe such heterostructure with a three dimensional [Formula: see text] Kane model, and use its eigenstates to derive an effective 2D Hamiltonian for the system. From these we obtain a phase diagram as a function of the well and barrier widths and we identify the different topological phases composed by zero, one, two, and three sets of edge states hybridized along the quantum wells. The phase transitions are characterized by a change of the spin Chern numbers and their corresponding band inversions. Complementary, transport measurements are experimentally investigated on a sample close to the transition line between the phases with one and two sets of edges states. Accordingly, for this sample we predict a gapless spectrum with low energy bulk conduction subbands given by one parabolic and one Dirac subband, and with edge states immersed in the bulk valence subbands. Consequently, we show that under these conditions, local and non-local transport measurements are inconclusive to characterize a sole edge state conductivity due to bulk conductivity. On the other hand, Shubnikov-de Haas (SdH) oscillations show an excellent agreement with our theory. Particularly, we show that the measured SdH oscillation frequencies agrees with our model and show clear signatures of the coexistence of a parabolic and Dirac subbands.

Somatic homozygous loss of SH2B3, and a non-Robertsonian translocation t(15;21)(q25.3;q22.1) with NTRK3 rearrangement, in an adolescent with progenitor B-cell acute lymphoblastic leukemia with the iAMP21.

Intrachromosomal amplification of chromosome 21 (iAMP21) occurs in ∼2% of B-cell acute lymphoblastic leukemia (ALL) and is considered to confer a poor prognosis. The relapse risk is associated with therapy intensity, suggesting that other somatic mutations may influence iAMP21-ALL prognosis. This abnormality is characterized by multiple copies of the RUNX1 gene in chromosome 21 and appears to arise through multiple breakage-fusion bridge cycles and chromothripsis. Rob(15;21) or a ring chromosome 21 have been associated with an increased risk for iAMP21-ALL, suggesting that constitutional genetic abnormalities may also drive leukemogenesis. Here we describe homozygous deletion of the SH2B3 gene, chromothripsis of chromosome 21, and a non-Robertsonian somatic t(15;21)(q25.3;q22.1) with NTRK3 gene rearrangement in an adolescent with iAMP21-B-ALL. Molecular cytogenetic studies detected iAMP21 with aCGH analysis revealing further genomic imbalances. The RT-qPCR analysis detected elevated expression levels of RUNX1 (68-fold) and reduced expression of CDK6 (0.057-fold). Studies with constitutive cells collected from mouth swabs showed that SH2B3 biallelic deletion was a somatic alteration occurring during clonal evolution. The identification of novel secondary genetic changes was valuable to discuss sporadic iAMP21 leukemogenic mechanisms. For the first time, we show a t(15;21)(q25.3;q22.1) with NTRK3 rearrangement in an adolescent with iAMP21-ALL.

Oxidative and osmotolerant effects in Salvator merianae (Squamata: Teiidae) red blood cells during hibernation.

Hibernation is a natural condition of animals that lives in the temperate zone, although some tropical lizards also experience hibernation annually, such as the lizard native from South America, Salvator merianae, or "tegu" lizard. Even though physiological and metabolic characteristic associated with hibernation have been extensively studied, possible alterations in the red blood cells (RBC) integrity during this period remains unclear. Dehydration and fasting are natural consequences of hibernating for several months and it could be related to some cellular modifications. In this study, we investigated if the osmotic tolerance of RBCs of tegu lizard under hibernation is different from the cells obtained from animals while normal activity. Additionally, we indirectly investigated if the RBCs membrane of hibernating tegus could be associated with oxidation by quantifying oxidized biomolecules and the activity of antioxidant enzymes. Our findings suggest that RBCs are more fragile during the hibernation period, although we did not find evidence of an oxidative stress scenario associated with the accentuated fragility. Even though we did not exclude the possibility of oxidative damage during hibernation, we suggested that an increased RBCs volume as a consequence of hypoosmotic blood during hibernation could also affect RBCs integrity as noted.

Caffeine intake reduces sedentary time and increases physical activity predisposition in obese police officers.

Sedentary time is associated with increased obesity in police officers. Caffeine intake may reduce sedentary time but it has not been extensively investigated. In the present study, the effect of caffeine ingestion on sedentary time was investigated in obese police officers. Fourteen obese police officers ingested either 5 mg/kg of caffeine or cellulose (placebo) for six days. Information on inactivity time, time spent with physical activities, self-reported perception of tiredness, and physical activity disposition was obtained daily during the intervention period. Sedentary and physical activity times were divided into two intraday periods (T1: 08:00 am-02:00 pm and T2: 02:00 pm-08:00 pm). Caffeine intake decreased the sedentary time in both T1 (79.2±2.2%) and T2 (79.1±2.5%), when compared with T2 of the placebo condition (81.1±3.6%, P<0.05). Caffeine intake also increased the time spent on light physical activities in T1 and T2 (17±2 and 18±2%), when compared with T2 of the placebo condition (16±3%, P<0.05). In addition, sedentary time increased and light physical activity time decreased from T1 to T2 in the placebo (P<0.001) but not in the caffeine condition (P=0.81). Caffeine intake had no effect on tiredness (P>0.05), but it increased the self-reported physical activity disposition compared to the placebo condition (4.5±2.7 vs 3.2±2.3 units, P<0.05). Caffeine intake reduced the sedentary time and increased the time spent on light physical activities of obese police officers, which seems to be related to a higher disposition for the practice of physical activity.

Functional analysis of ldlr genetic variants identified in hypercholesterolemic patients in brazil, USING CRISPR/CAS9

BACKGROUND AND AIMS: Introduction: The familial hypercholesterolemia (FH) is one of the main causes of cardiovascular diseases, and it is mainly caused by genetic variants at the low-density lipoprotein receptor (LDLR). Although ultrasequencing technology has allowed the identification of several genetic variants, few of them was functional analyzed. The CRISPR/ Cas9 tool promotes precise genetic editing and allows the creation of experimental models, therefore contributing to the functional validation process. Aim: To use the CRISPR/Cas9 tool to perform in vitro functional analysis of LDLR variants identified in FH patients. METHODS: Two missense LDLR variants were selected within a group of variants identified in FH patients, based on in silico data, the affected protein domain and MAF. Three sgRNAs were designed for each of the variants c.551G>A and c.1118G>A, to analyze the accuracy of the sgRNAs. The sgRNAs were inserted on PX458 plasmid, cloned, purified in E. coli DH5a, and then co-transfected with the DNA template at HepG2 cells. The DNAs templates were designed to contain the selected variants. RESULTS: HepG2 cells co-transfected with PX458 constructs and DNA templates showed considerably transfection rate, being possible to visualize it at fluorescence microscopy. However, it was noted that single transfection of sgRNAs showed a higher transfection efficiency than cotransfection. CONCLUSIONS: We designed sgRNA for c.551G>A and c.1118G>A variants, being able to analyze the transfection efficiency. In further steps, we will select new sgRNAs for LDLR variants that have not been described yet, and functional analysis will be performed to determine the clinical relevance of these variants.

Anti-staphylococcal effects of Myracrodruon urundeuva lectins on nonresistant and multidrug resistant isolates.

AIMS: To evaluate the anti-staphylococcal effects of lectins isolated from bark (MuBL), heartwood (MuHL) and leaves (MuLL) of Myracrodruon urundeuva. METHODS AND RESULTS: The lectins were evaluated for: effects on growth, aggregation, haemolytic activity and biofilm-forming ability of Staphylococcus aureus clinical isolates nonresistant (8325-4) and multidrug resistant (LAC USA300); interference with the expression of virulence genes (hla, rnaIII and spa) of the Agr system of S. aureus; and synergistic effect with the antibiotics cefoxitin and cefotaxime. MuBL, MuHL and MuLL reduced growth (minimal inhibitory concentration (MIC): 12·5-50 µg ml-1 ) and viability (minimal bactericidal concentration (MBC): 100 µg ml-1 ) of 8325-4 and LAC USA300 cells. MuLL (at ½MIC and MIC) reduced LAC USA300 agglutination. The lectins did not interfere with haemolytic activity and expression of hla, rnaIII and spa genes. Only MuHL was able to reduce the biofilm production by 8325-4 (50-400 µg ml-1 ) and LAC USA300 (400 µg ml-1 ). CONCLUSION: The M. urundeuva lectins showed antibacterial activity against nonresistant and resistant clinical isolates of S. aureus and synergistic effects with antibiotics in reducing growth and biofilm formation. SIGNIFICANCE AND IMPACT OF THE STUDY: This work reports bioactive molecules capable of acting as anti-staphylococcal agents, since there are increasing reports of multiresistant isolates of this bacterium.

The use of outdoor gyms is associated with women and low-income people: a cross-sectional study.

OBJECTIVE: To examine the prevalence, and the demographic, socio-economic, and health correlates to Outdoor Gyms (OGs) use for adults from a southern Brazilian city. STUDY DESIGN: Population-based cross-sectional study. METHODS: A total of 431 adults (66.8% women) aged 18-87 years living in the surroundings of four OGs distributed in different regions of the city were randomly selected. Information about OG use for physical activity (PA) practice, and demographic, socio-economic, and health variables were collected by household interviews. Associations between independent variables and OG use were analyzed with results expressed as odds ratio (OR) and 95% confidence interval (95% CI). RESULTS: About one-third of participants (30.4%; 95% CI: 26.1-34.7) informed using OGs for PA practice, and 20.4% (95% CI 16.8-24.4) informed using it twice or more times a week (≥2x/week). Adjusted analysis indicated that the OG use ≥2x/week is higher for women (OR: 1.93; 95% CI: 1.11-3.35) and for those with lower family income (OR: 2.13; 95% CI: 1.03-4.13) than men and those with higher family income, respectively. CONCLUSION: About 30% of the population uses OGs for PA practice. Women and low-income people are those who more commonly use OGs for PA practice. The installation of these facilities in public spaces may reduce social inequities related to leisure-time PA.

Antibacterial effects of the lectin from pomegranate sarcotesta (PgTeL) against Listeria monocytogenes.

AIMS: To investigate the effects of the lectin from Punica granatum sarcotesta (PgTeL) on growth, viability, cell structure, biofilm formation and chitinase activity of Listeria monocytogenes. In addition, the effect of PgTeL on the adhesion and invasion of human cells (HeLa) was determined. METHODS AND RESULTS: PgTeL showed bacteriostatic and bactericidal effects on the strains L. monocytogenes N53-1 and EGD-e, causing morphometric alterations, cell aggregation, strong deformation and cell disruption. PgTeL inhibited biofilm formation by EGD-e and N53-1 and also interfered with the adhesion and invasion processes of EGD-e and N53-1 in HeLa cells. Finally, the chitinase activity of L. monocytogenes EGD-e was reduced in the presence of PgTeL, which can be involved in the inhibition of adhesion process. CONCLUSION: PgTeL is an antibacterial agent against L. monocytogenes, inhibiting growth and promoting cell death, as well as impairing biofilm formation and bacterial adhesion and invasion into human cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The results stimulate future investigations on the potential of PgTeL for protection of contamination in food products.

Caffeine intake reduces sedentary time and increases physical activity predisposition in obese police officers

Sedentary time is associated with increased obesity in police officers. Caffeine intake may reduce sedentary time but it has not been extensively investigated. In the present study, the effect of caffeine ingestion on sedentary time was investigated in obese police officers. Fourteen obese police officers ingested either 5 mg/kg of caffeine or cellulose (placebo) for six days. Information on inactivity time, time spent with physical activities, self-reported perception of tiredness, and physical activity disposition was obtained daily during the intervention period. Sedentary and physical activity times were divided into two intraday periods (T1: 08:00 am-02:00 pm and T2: 02:00 pm-08:00 pm). Caffeine intake decreased the sedentary time in both T1 (79.2±2.2%) and T2 (79.1±2.5%), when compared with T2 of the placebo condition (81.1±3.6%, P<0.05). Caffeine intake also increased the time spent on light physical activities in T1 and T2 (17±2 and 18±2%), when compared with T2 of the placebo condition (16±3%, P<0.05). In addition, sedentary time increased and light physical activity time decreased from T1 to T2 in the placebo (P<0.001) but not in the caffeine condition (P=0.81). Caffeine intake had no effect on tiredness (P>0.05), but it increased the self-reported physical activity disposition compared to the placebo condition (4.5±2.7 vs 3.2±2.3 units, P<0.05). Caffeine intake reduced the sedentary time and increased the time spent on light physical activities of obese police officers, which seems to be related to a higher disposition for the practice of physical activity.