A Thermoresponsive injectable drug delivery system of chitosan/ß-glycerophosphate with gellan gum/alginate microparticles.

The development of new Drug Delivery Systems (DDS) by incorporating microparticles within hydrogels can prolong the release rate of drugs and/or other bioactive agents. In this study, we combined gellan gum/alginate microparticles within a thermoresponsive chitosan (Ch) hydrogel with ß-Glycerophosphate (ß-GP), designing the system to be in the sol state at 21 °C and in the gel state at 37 °C to enable the injectability of the system. The system was in the sol state between 10 °C and 21 °C. Higher concentrations of ß-GP (0, 2, 3, 4, 5 w/v%) and microparticles (0, 2 and 5 w/v%) allowed a faster sol-gel transition with higher mechanical strength at 37 °C. However, the sol-gel transition was not instantaneous. The release profile of methylene blue (MB) from the microparticles was significantly affected by their incorporation in Ch/ß-GP hydrogels, only allowing the release of 60-70 % of MB for 6 days, while the microparticles alone released all the MB in 48 h. The proposed system did not present cytotoxicity to VERO cell lines as a preliminary assay, with the Ch/ß-GP/GG:Alg having >90 % of cellular viability. The proposed Ch/ß-GP system proved to have a delaying effect on drug release and biocompatible properties, being a promising future DDS.

Evolution of guanylate binding protein genes shows a remarkable variability within bats (Chiroptera).

Background: GBPs (guanylate binding proteins), an evolutionary ancient protein family, play a key role in the host's innate immune response against bacterial, parasitic and viral infections. In Humans, seven GBP genes have been described (GBP1-7). Despite the interest these proteins have received over the last years, evolutionary studies have only been performed in primates, Tupaia and rodents. These have shown a pattern of gene gain and loss in each family, indicative of the birth-and-death evolution process. Results: In this study, we analysed the evolution of this gene cluster in several bat species, belonging to the Yangochiroptera and Yinpterochiroptera sub-orders. Detailed analysis shows a conserved synteny and a gene expansion and loss history. Phylogenetic analysis showed that bats have GBPs 1,2 and 4-6. GBP2 has been lost in several bat families, being present only in Hipposideidae and Pteropodidae. GBPs1, 4 and 5 are present mostly as single-copy genes in all families but have suffered duplication events, particularly in Myotis myotis and Eptesicus fuscus. Most interestingly, we demonstrate that GBP6 duplicated in a Chiroptera ancestor species originating two genes, which we named GBP6a and GBP6b, with different subsequent evolutionary histories. GBP6a underwent several duplication events in all families while GBP6b is present as a single copy gene and has been lost in Pteropodidae, Miniopteridae and Desmodus rotundus, a Phyllostomidae. With 14 and 15 GBP genes, Myotis myotis and Eptesicus fuscus stand out as having far more copies than all other studied bat species. Antagonistically, Pteropodidae have the lowest number of GBP genes in bats. Conclusion: Bats are important reservoirs of viruses, many of which have become zoonotic diseases in the last decades. Further functional studies on bats GBPs will help elucidate their function, evolutionary history, and the role of bats as virus reservoirs.

First report of Rhytidodes gelatinosus (Rudolphi, 1819) Looss, 1901 (Digenea: Rhytidodidae) in an olive-ridley Turtle Lepidochelys olivacea (Eschscholtz, 1829) from Brazil.

This article reports the first occurrence of Rhytidodes gelatinosus (Rudolphi, 1819) Looss, 1901 (Digenea: Rhytidodidae) in the olive-ridley turtle Lepidochelys olivacea (Testudines: Chelonidae), in an individual found in the State of Sergipe, Brazil. Although R. gelatinosus has already been described in other species of sea turtles in the world, this is the first report of this parasite in L. olivacea. We also present a list of hosts and locations where this helminth has already been identified.

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.

Brain glucose hypometabolism and hippocampal inflammation in Goto-Kakizaki rats.

Brain glucose hypometabolism and neuroinflammation are early pathogenic manifestations in neurological disorders. Neuroinflammation may also disrupt leptin signaling, an adipokine that centrally regulates appetite and energy balance by acting on the hypothalamus and exerting neuroprotection in the hippocampus. The Goto-Kakizaki (GK) rat is a non-obese type 2 diabetes mellitus (T2DM) animal model used to investigate diabetes-associated molecular mechanisms without obesity jeopardizing effects. Wistar and GK rats received the maintenance adult rodent diet. Also, an additional control group of Wistar rats received a high-fat and high-sugar diet (HFHS) provided by free consumption of condensed milk. All diets and water were provided ad libitum for eight weeks. Brain glucose uptake was evaluated by 2-deoxy-2-[fluorine-18] fluoro-D-glucose under basal (saline administration) or stimulated (CL316,243, a selective ß3-AR agonist) conditions. The animals were fasted for 10-12 h, anesthetized, and euthanized. The brain was quickly dissected, and the hippocampal area was sectioned and stored at -80°C in different tubes for protein and RNA analyses on the same animal. GK rats exhibited attenuated brain glucose uptake compared to Wistar animals and the HFHS group under basal conditions. Also, the hippocampus of GK rats displayed upregulated leptin receptor, IL-1ß, and IL-6 gene expression and IL-1ß and the subunit of the transcription factor NF-κB (p-p65) protein expression. No significant alterations were detected in the hippocampus of HFHS rats. Our data indicated that a genetic predisposition to T2DM has significant brain deteriorating features, including brain glucose hypometabolism, neuroinflammation, and leptin signaling disruption in the hippocampal area.

Quejas visuales durante la pandemia de COVID-19 en España y Portugal / Visual complaints during the COVID-19 pandemic in Spain and Portugal

Objetivo Este estudio reporta los hábitos de la población y las quejas oculares relacionadas con la salud visual en el contexto de la pandemia de COVID-19 a partir de visitas realizadas durante 2021 en España y en Portugal. Material y métodos Invitación por correo electrónico a una encuesta transversal online y también realizada en persona a pacientes de clínicas de oftalmología de España y de Portugal de septiembre a noviembre de 2021. Participaron 3.833 encuestados mayores de 18años con respuestas anónimas válidas. Resultados El 60% de los encuestados explicó mucha incomodidad causada por el aumento de los síntomas de ojo seco debido al trabajo digital más intenso y el empañamiento de los lentes al usar mascarillas. El 81,6% de los encuestados usaba dispositivos digitales al menos 3horas en promedio por día, y el 40% comenzó a usar dispositivos digitales más de 8horas en promedio por día. Además, el 44% de los encuestados sintió que su visión de cerca había empeorado en este período. El primer síntoma importante de la presbicia estaba relacionado con la dificultad para leer las letras más pequeñas de los paquetes. El 86% presentó los primeros síntomas a los 40años. Las ametropías más frecuentes identificadas fueron miopía (40,2%) y astigmatismo (36,7%). Para los padres, tener buena vista (87,2%) era el aspecto más valorado en la vida de sus hijos. Conclusiones Los hallazgos brindan una idea de los desafíos durante la COVID-19 para las prácticas oftalmológicas. En una sociedad altamente dependiente de la visión, es fundamental centrarse en los signos y los síntomas que conducen a afecciones oftalmológicas. El uso excesivo de dispositivos digitales y el uso de mascarillas durante esta pandemia han agravado algunos, señalando la importancia de la referencia para planificar una atención ocular eficiente en situaciones similares (AU)

Purpose This study reports visual health during the COVID-19 pandemic in 2021 in Spain and Portugal, focusing on eye complaints and population habits. Material and methods Cross-sectional survey through an online email invitation to patients attending ophthalmology clinics in Spain and Portugal from September to November 2021. Around 3833 participants offered valid anonymous responses in a questionnaire. Results Sixty percent of respondents identified significant discomfort related to dry eye symptoms for increased screen time and lens fogging using facemasks. 81.6% of the participants used digital devices for more than 3h per day and 40% for more than 8h. In addition, 44% of participants referred to worsening near vision. The most frequent ametropias were myopia (40.2%) and astigmatism (36.7%). Parents considered eyesight the most important aspect of their children (87.2%). Conclusions The results show the challenges for eye practices during the initial COVID-19 pandemic. Focusing on signs and symptoms that lead to ophthalmologic conditions is an essential concern, especially in our digital society highly dependent on vision. At the same time, the excessive use of digital devices during this pandemic has aggravated dry eye and myopia (AU)

Visual complaints during the COVID-19 pandemic in Spain and Portugal.

PURPOSE: This study reports visual health during the COVID-19 pandemic in 2021 in Spain and Portugal, focusing on eye complaints and population habits. MATERIAL AND METHODS: Cross-sectional survey through an online email invitation to patients attending ophthalmology clinics in Spain and Portugal from September to November 2021. Around 3833 participants offered valid anonymous responses in a questionnaire. RESULTS: Sixty percent of respondents identified significant discomfort related to dry eye symptoms for increased screen time and lens fogging using facemasks. 81.6% of the participants used digital devices for more than 3 h per day and 40% for more than 8 h. In addition, 44% of participants referred to worsening near vision. The most frequent ametropias were myopia (40.2%) and astigmatism (36.7%). Parents considered eyesight the most important aspect of their children (87.2%). CONCLUSIONS: The results show the challenges for eye practices during the initial COVID-19 pandemic. Focusing on signs and symptoms that lead to ophthalmologic conditions is an essential concern, especially in our digital society highly dependent on vision. At the same time, the excessive use of digital devices during this pandemic has aggravated dry eye and myopia.

[Visual complaints during the COVID-19 pandemic in Spain and Portugal]. / Quejas visuales durante la pandemia de COVID-19 en España y Portugal.

Purpose: This study reports visual health during the COVID-19 pandemic in 2021 in Spain and Portugal, focusing on eye complaints and population habits. Material and methods: Cross-sectional survey through an online email invitation to patients attending ophthalmology clinics in Spain and Portugal from September to November 2021. Around 3833 participants offered valid anonymous responses in a questionnaire. Results: Sixty percent of respondents identified significant discomfort related to dry eye symptoms for increased screen time and lens fogging using facemasks. 81.6% of the participants used digital devices for more than 3 h per day and 40% for more than 8 h. In addition, 44% of participants referred to worsening near vision. The most frequent ametropias were myopia (40.2%) and astigmatism (36.7%). Parents considered eyesight the most important aspect of their children (87.2%). Conclusions: The results show the challenges for eye practices during the initial COVID-19 pandemic. Focusing on signs and symptoms that lead to ophthalmologic conditions is an essential concern, especially in our digital society highly dependent on vision. At the same time, the excessive use of digital devices during this pandemic has aggravated dry eye and myopia.

Effect of Activated Charcoal Toothpaste on Color Stability of Bleached Teeth Immersed in Different Drinks.

OBJECTIVE: This study aimed to evaluate the effect of activated charcoal toothpaste on the color stability of teeth subjected to tooth bleaching and pigmenting agents. METHODS: A total of 120 bovine crowns were randomly divided into 12 groups (n=10) according to two study factors: staining solutions (three levels): saliva (control), coffee, and red wine; and toothpaste (four levels): BPC, Bianco Pro Clinical (Bianco Oral Care) (Control); BIW, Black is White (Curaprox); BCA, Bianco Carbon (Bianco Oral Care); and NAT, Natural Suavetex (Suavetex). The samples were subjected to office bleaching with a 35% hydrogen peroxide-based gel (Whiteness HP Blue, FGM), followed by immersion in the solution for 45 minutes per day and daily toothbrushing for 7 days. The color (ΔE) and luminosity changes (ΔL*) were measured using reflectance spectroscopy (Vita EasyShade). The CIE values (L*, a*, b*) were measured at baseline after bleaching (T0) and immediately after immersion in solution each day (Ti1-Ti7) and after all toothbrushing cycles (Tb1-Tb7). ΔE and ΔL were analyzed using a two-way analysis of variance and Tukey's test (α=0.05). The clinically unacceptable level of ΔE > 3.3 was used to evaluate the color change. RESULTS: The color change was significantly influenced by the staining solutions and toothpastes (p<0.001). The color change (ΔE) was significantly higher when immersed in wine than in coffee, and lower ΔE values were observed for artificial saliva (control), irrespective of the toothpaste used. In artificial saliva, BPC, BIW, and BCA resulted in significantly lower ΔE values than NAT, which presented a clinically unacceptable level of dental color change (ΔE>3.3). Coffee resulted in a lower (L*) reduction than wine, irrespective of the toothpaste used. CONCLUSION: Charcoal toothpastes resulted in a color change on the surface of the tooth enamel (ΔE). The bleaching effect of the charcoal toothpastes and control evaluated in this study partially reduced the color changes on the surface of the tooth enamel caused by staining solutions but was unable to reestablish the measured values to the baseline. For teeth immersed in artificial saliva, the color change was not noticeable in BCA, BIW, and control-BPC (ΔE≥3.3), except for NAT, which showed a significant color change.

Brain glucose hypometabolism and hippocampal inflammation in Goto-Kakizaki rats

Brain glucose hypometabolism and neuroinflammation are early pathogenic manifestations in neurological disorders. Neuroinflammation may also disrupt leptin signaling, an adipokine that centrally regulates appetite and energy balance by acting on the hypothalamus and exerting neuroprotection in the hippocampus. The Goto-Kakizaki (GK) rat is a non-obese type 2 diabetes mellitus (T2DM) animal model used to investigate diabetes-associated molecular mechanisms without obesity jeopardizing effects. Wistar and GK rats received the maintenance adult rodent diet. Also, an additional control group of Wistar rats received a high-fat and high-sugar diet (HFHS) provided by free consumption of condensed milk. All diets and water were provided ad libitum for eight weeks. Brain glucose uptake was evaluated by 2-deoxy-2-[fluorine-18] fluoro-D-glucose under basal (saline administration) or stimulated (CL316,243, a selective β3-AR agonist) conditions. The animals were fasted for 10-12 h, anesthetized, and euthanized. The brain was quickly dissected, and the hippocampal area was sectioned and stored at -80°C in different tubes for protein and RNA analyses on the same animal. GK rats exhibited attenuated brain glucose uptake compared to Wistar animals and the HFHS group under basal conditions. Also, the hippocampus of GK rats displayed upregulated leptin receptor, IL-1β, and IL-6 gene expression and IL-1β and the subunit of the transcription factor NF-κB (p-p65) protein expression. No significant alterations were detected in the hippocampus of HFHS rats. Our data indicated that a genetic predisposition to T2DM has significant brain deteriorating features, including brain glucose hypometabolism, neuroinflammation, and leptin signaling disruption in the hippocampal area.

Brain glucose hypometabolism and hippocampal inflammation in Goto-Kakizaki rats

Brain glucose hypometabolism and neuroinflammation are early pathogenic manifestations in neurological disorders. Neuroinflammation may also disrupt leptin signaling, an adipokine that centrally regulates appetite and energy balance by acting on the hypothalamus and exerting neuroprotection in the hippocampus. The Goto-Kakizaki (GK) rat is a non-obese type 2 diabetes mellitus (T2DM) animal model used to investigate diabetes-associated molecular mechanisms without obesity jeopardizing effects. Wistar and GK rats received the maintenance adult rodent diet. Also, an additional control group of Wistar rats received a high-fat and high-sugar diet (HFHS) provided by free consumption of condensed milk. All diets and water were provided ad libitum for eight weeks. Brain glucose uptake was evaluated by 2-deoxy-2-[fluorine-18] fluoro-D-glucose under basal (saline administration) or stimulated (CL316,243, a selective β3-AR agonist) conditions. The animals were fasted for 10-12 h, anesthetized, and euthanized. The brain was quickly dissected, and the hippocampal area was sectioned and stored at -80°C in different tubes for protein and RNA analyses on the same animal. GK rats exhibited attenuated brain glucose uptake compared to Wistar animals and the HFHS group under basal conditions. Also, the hippocampus of GK rats displayed upregulated leptin receptor, IL-1β, and IL-6 gene expression and IL-1β and the subunit of the transcription factor NF-κB (p-p65) protein expression. No significant alterations were detected in the hippocampus of HFHS rats. Our data indicated that a genetic predisposition to T2DM has significant brain deteriorating features, including brain glucose hypometabolism, neuroinflammation, and leptin signaling disruption in the hippocampal area.

Death and non-lethal weapons: A case of homicide by penetrating injury without projectile.

Herein, we report the death of a man, approximately thirty years old, victim of a short-range shot to the thorax from a PIEXON JPX4, a hand weapon classed category D in France, said to be non-lethal. External examination of the lesion revealed characteristics similar to those of ballistic injuries. The autopsy found an intercostal wound that reached the pulmonary parenchyma and the pulmonary artery, with severe hemothorax. By the end of the autopsy, no projectile had been found in the body, nor any exit orifice. Death was caused by the effects of hemorrhagic shock. The PIEXON JPX4 has four cartridges, projecting a capsaicin gel that is designed to irritate the ENT area (ear nose throat) to incapacitate an assailant. The manufacturer recommends not to use it at distances of less than 1.5 m. Experimental shots were performed on gelatine blocks at point-blank range and at distances of 5, 10, 20, 30, 50, 100 and 150 cm to evaluate the distance necessary for the jet of gel to have a penetrating effect. Shots at 5-30 cm penetrated the structure. None of the other shots were penetrating. The autopsy and experiment data therefore show the penetrating potential of the jet of gel. Herein, we report the first death due to use of the PIEXON JPX4. From a forensic investigation viewpoint, we add a new exception to the "bullet rule". (The odd and even bullet rule states that if the number of gunshot wounds of entrance and exit found in the body is even, the presumption is that no bullet is lodged in the body. If the number of gunshot wounds of entrance and exit is odd, the presumption is that one or more bullets have been lodged in the body.).

Crude extract of Jatobá leaves promotes canine osteosarcoma cell D17 proliferation.

Background and Aim: New substances for neoplasm treatment have to be carefully studied to minimize adverse effects and prevent disease progression stimulation. Jatobá is a typical tree of the Cerrado and Caatinga biome, with antifungal, antimicrobial, larvicide, antioxidant, and antiproliferative properties. This study aimed to investigate the action of the crude extract of Jatobá leaves (EBFJ) on canine osteosarcoma (CO) cells and analyze the expression of biomarkers in neoplasm progression. Materials and Methods: D17 cells were cultured and subjected to treatment with EBFJ at different concentrations (10 µg/mL; 100 µg/mL; 1000 µg/mL; 2000 µg/mL; and 5000 µg/mL) and exposure times (24 h, 48 h, and 72 h). The tetrazolium reduction assay and the immunocytochemistry technique, with anti-Bcl2, anti-p53, and anti-Ki-67 antibodies, were used to observe the effect of the extract on cell proliferation. Results: Doses of 2000 µg and 5000 µg had cell viability of 300.80% and 361.84%, respectively. The extract did not show significant cytotoxicity of samples with the control group. The confluence of cells, the number of labeled cells, and the expression of Bcl2, Ki-67, and p53 were higher in the groups treated with EBFJ, with a statistical difference from the group without treatment. Conclusion: EBFJ was not cytotoxic and had a proliferative effect on CO D17 cells. The confluence of cells, the number of labeled cells, and the expression of Bcl2, Ki-67, and p53 were higher in the groups treated with the extract.

Effects of Charcoal Toothpaste on the Surface Roughness, Color Stability, and Marginal Staining of Resin Composites.

OBJECTIVE: This study was designed to evaluate the effects of charcoal toothpaste on the surface roughness, color stability, and marginal staining of resin composite restorations. METHODS: A total of 100 bovine incisors was collected. The crowns were sectioned and randomly divided into 10 groups (n=10) according to two study factors: toothpaste groups and nanoparticle resin composite groups. Five toothpastes-Bianco Pro Clinical (Bianco Oral Care, Uberlândia, MG, Brazil) - Control group; Bianco Carbon (Bianco Oral Care); NAT, Natural Suavetex Carvão Ativado (Suavetex, Uberlândia, MG, Brazil); Nano Action Black Be Emotion (Polishop, Jundiaí, SP, Brazil); and BIW, Black is White (Curaprox, Curaden AG, Kriens, Switzerland)-and two resin composites-Z350XT (Filtek Z350XT, 3M Oral Care) and Vittra (Vittra APS FGM, Joinville, SC, Brazil)-were used. Circular cavities with a diameter of 4 mm and a depth of 1 mm were prepared on the buccal face of the tooth crowns and restored with resin composites. The specimens were subjected to three months of simulated toothbrushing. The surface roughness (right angle [Ra], in micrometers [µm]) of the resin composites was measured before and after toothbrushing in five areas per specimen. The resin composite color and luminosity changes (ΔE and ΔL, respectively) were measured using reflectance spectroscopy (Vita EasyShade). Macro photographs were taken before and after toothbrushing to qualitatively analyze the marginal staining (MSt) of the resin composite restorations. Scanning electron microscopy (SEM) was performed before and after the simulated toothbrushing. Ra data were analyzed using two-way analysis of variance with repeated measures and the Tukey HSD test; MSt was analyzed using Kruskal-Wallis and Dunn tests (α=0.05), and the resin composite color change was analyzed using the clinically unacceptable level of ΔE > 3.3. RESULTS: Simulated brushing increased Ra irrespective of the resin composite or toothpaste used. No significant differences were found in Ra between the control group and all groups on which the charcoal toothpastes were tested. A clinically unacceptable level of resin composite color change (ΔE>3.3) was found after the use of most charcoal toothpastes. Use of Bianco Carbon resulted in marginal staining similar to that of the control group and was lower than that of the other charcoal toothpastes. Vittra brushed with black toothpaste showed the highest marginal staining. CONCLUSION: Use of charcoal toothpaste resulted in Ra values of resin composites similar to those found with conventional toothpastes. Charcoal toothpaste generally resulted in clinical resin composite color changes (ΔE). All charcoal toothpastes, except Bianco Carbon, caused marginal staining of the resin composite restorations.

Variante no gene LDL-R associada com elevação do LDL-C identificada em um portador de hipercolesterolemia familiar / LDL-R gene variant associated with LDL-C elevation identified in a carrier of familial hypercholesterolemia

INTRODUÇÃO: A hipercolesterolemia familiar (HF) é uma doença hereditária autossômica codominante marcada por altos níveis de colesterol ligado à lipoproteína de baixa densidade (LDL-c). Pode-se apresentar na forma homozigótica, apresentando prevalência de 1/160.000 a 1/1.000.000 ou na forma heterozigótica, um caso em cada 200-250 pessoas. Material e MÉTODO: Paciente do sexo feminino, aos 11 meses foi encaminhada ao setor de Dislipidemia para acompanhamento. Na primeira consulta, apresentava colesterol total (CT): 325 mg/dL, LDL-c: 255 mg/dL, colesterol ligado à lipoproteína de alta densidade (HDL-c): 44 mg/dL, triglicerídeos (TG): 128 mg/ dL. Exame físico: sem alterações. Durante o seguimento, paciente realizou doppler de carótidas e vertebrais sem aumento da espessura médio intimal e tomografia computadorizada com escore de cálcio de zero. Paciente foi incluída no protocolo para o sequenciamento exômico dos principais genes relacionados com a HF. A paciente apresentou 9 diferentes variantes, todos em heterozigose, com cobertura maior que 30 vezes, distribuídas nos 3 genes, no entanto nenhuma previamente associada com HF. RESULTADOS: Trata-se de uma criança cuja alteração do perfil lipídico foi identificada precocemente. A análise do perfil genético identificou uma mutação que está relacionada ao fenótico de HF. As variantes mais comuns relacionadas à HF estão localizadas no gene que codifica o receptor da LDL (LDLR). Dentre as variantes analisadas, somente a variante rs1433099 no gene LDLR foi previamente associada a elevação do LDL-c e maior suscetibilidade a desenvolver doença arterial coronariana. No entanto esse achado nunca havia sido descrito em pacientes com diagnóstico clínico de HF. DISCUSSÃO E CONCLUSÕES: O relato de caso apresentou uma variante genética que não havia sido descrita previamente relacionada à HF. Dessa forma, devido à grande importância da hereditariedade nos quadros de dislipidemias, à pequena parcela de genes descritos e validados, sugere-se que a variante rs1433099 no gene LDLR seja incorporada nos exames genéticos associadas a essa patologia; possibilitando a expansão do diagnóstico, além do tratamento precoce dessa enfermidade tão frequente.

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.

Histological classification and expression of markers of canine mast cell tumors.

BACKGROUND AND AIM: Mast cell tumors (MCTs) are malignant neoplasms that are common in dogs. Their biological behavior is variable and unpredictable. The aim of the present study was to analyze the histological classification and expression of markers of canine MCTs. MATERIALS AND METHODS: Thirty samples of canine MCTs were graded according to the histological classification methods of Patnaik and those of Kiupel. The expression of phosphoprotein 53 (p53) and c-kit proteins was quantified by immunohistochemistry using image processing software, ImageJ - a public domain computer program, developed at the National Institutes of Health. RESULTS: It was possible to determine the grade of 100% of the samples. According to Patnaik's classification, 20.00% of the samples were Grade 1, 43.30% were Grade 2, and 36.70% were Grade 3. According to Kiupel's classification, 56.67% of the samples were of high intensity and 43.33% were of low intensity. Grade 1 tumors had the highest expression of p53 and c-kit, and Grade 2 had the lowest expression. The results showed that it is necessary to perform both histological grading methods. The classification into high and low intensity may provide more consistent results than the three-level grading system. However, a smaller number of categories, although it facilitates the classification, may not be sufficient for the prognosis. CONCLUSION: Quantitative evaluation of p-53 and c-kit expression is a useful tool to increase the accuracy of the analysis and to aid in choosing the treatment method for canine MCTs. Histological grading should be combined with other diagnostic methods.

Hydrocarbon Contamination Patterns in the Cenotes of the Mexican Caribbean: The Application of Principal Component Analysis.

Cenotes are naturally occurring flooded caves that are frequent in Mexico's Yucatan Peninsula; they result from the collapse of limestone bedrock into the regional groundwater table. Cenotes in Quintana Roo are important ecological and economic hot spots but are susceptible to anthropogenic pollution. In this study, we collected water samples from 11 cenotes over multiple years to evaluate polycyclic aromatic hydrocarbon (PAH) concentrations and patterns as associated with tourist traffic. The primary PAHs detected in samples included fluoranthene, anthracene, phenanthrene and naphthalene, with total PAH concentrations increasing almost fivefold for cenotes sampled from 2016 to 2017. This is compared to only a 7% increase in tourist traffic during these years. Multivariate statistical analysis of the PAH concentration data suggests that diesel, gasoline and asphalt are the most likely pollution sources and that they are associated with periods of increased tourist traffic.

Short communication: Socio-psychological factors influencing dairy farmers' intention to adopt high-grain feeding in Brazil.

This study aimed to apply the theory of planned behavior to determine the effects of attitude, perceived behavioral control, and subjective norms on the intention of dairy farmers toward high-grain feeding. Quantitative data were collected through interviews with 150 dairy farmers in Paraná, Brazil. Partial least squares structural equation modeling was used to analyze the data. We observed that farmers with a positive perception of grain feeding (attitude) and a positive perception of their ability to increase the use of grains (perceived behavioral control) had higher intention to increase grain feeding. Social pressure (subjective norm) had no effect on the intention to grain feed. These results can contribute to the development of public strategies and policies that encourage the use of high-grain diets in dairy farms.

Eficácia da suplementação oral com 1, 3-1, 6 betaglucano proveniente de Saccharomyces cerevisiae no controle da mastite bovina / [Efficiency of oral supplementation of 1. 3-1. 6 beta-glucan from saccharomyces cerevisiae on the control of bovine mastitis]

A mastite bovina, uma das principais doenças do rebanho leiteiro, caracteriza-se por um processo inflamatório no úbere. A inviabilidade econômica, o impacto ambiental negativo e os resíduos antimicrobianos têm estimulado a pesquisa de outros tratamentos alternativos para a prevenção e o tratamento de doenças na bovinocultura leiteira. O betaglucano é um agente imunomodulador com potencial ação preventiva para doenças infecciosas, inclusive a mastite. Este estudo teve como objetivo avaliar a eficácia do uso do betaglucano, por meio de administração oral, em animais em lactação. Foram utilizadas 20 vacas lactantes, distribuídas em dois grupos, um controle e um tratamento, com 10 animais em cada grupo. O grupo tratamento recebeu 5g/dia, durante 60 dias, de 1,3-1,6 betaglucano isolado da parede celular de Saccharomyces cerevisiae diluído em ração após a ordenha, enquanto o grupo controle recebia somente a ração. Foram realizados os testes de California Mastitis Test (CMT), contagem de células somáticas (CCS), produção de leite e percentual de gordura e proteína no leite. Não houve diferença estatisticamente significativa entre os grupos quanto à CCS, ao CMT, à composição do leite ou produção. Não se observou, portanto, eficácia do uso do betaglucano purificado, administrado por via oral, no controle e na prevenção da mastite em vacas leiteiras, quando comparadas com o grupo controle. Atribuem-se esses resultados, principalmente, à degradação ruminal do produto testado. Sugerem-se, portanto, mais pesquisas utilizando o 1,3-1,6 betaglucano purificado de parede de S. cerevisiae por outras vias de administração, tais como intramamária e subcutânea.(AU)

Bovine mastitis, one of the main diseases of dairy herds, is characterized by an inflammatory process in the udder. The economic and environmental impacts, as well as the residues of antimicrobial drugs have stimulated the research of novel alternative treatments for the prevention and treatment of diseases in dairy production cows. The beta-glucan is an immunomodulator agent, with potential preventive action for infectious diseases, including mastitis. This study aimed to assess the effectiveness of orally administered beta-glucan in lactating cows. 20 lactating cows were used, distributed into two groups, one control and one treatment, with 10 cows in each group. The treatment group received 5g of 1.3-1.6 betaglucan daily for 60 days, isolated from the cell-wall of Saccharomyces cerevisiae diluted into a grain meal, whereas the animals in the control group received only the ration. The California Mastitis Test (CMT), Somatic Cells Counting (SCC), daily production and assessments of fat and protein content in milk were done. There was no statistically significant difference between the groups concerning subclinical mastitis detected by CMT, SCC, milk production and composition regarding protein and fat content. It was not observed, therefore, the effectiveness of the use of purified beta-glucan orally administered on the control or prevention of mastitis in dairy cows. The results are attributed to the ruminal degradation of the product tested. It is, therefore, suggested that more research should be conducted using the 1.3-1.6 beta-glucan purified from the cell wall of S. cerevisiae by other administration means and ruminal protection technologies for the isolated beta-glucan.(AU)