A three-dimensional valve-on-chip microphysiological system implicates cell cycle progression, cholesterol metabolism and protein homeostasis in early calcific aortic valve disease progression.

BACKGROUND: Calcific aortic valve disease (CAVD) is one of the most common forms of valvulopathy, with a 50 % elevated risk of a fatal cardiovascular event, and greater than 15,000 annual deaths in North America alone. The treatment standard is valve replacement as early diagnostic, mitigation, and drug strategies remain underdeveloped. The development of early diagnostic and therapeutic strategies requires the fabrication of effective in vitro valve mimetic models to elucidate early CAVD mechanisms. METHODS: In this study, we developed a multilayered physiologically relevant 3D valve-on-chip (VOC) system that incorporated aortic valve mimetic extracellular matrix (ECM), porcine aortic valve interstitial cell (VIC) and endothelial cell (VEC) co-culture and dynamic mechanical stimuli. Collagen and glycosaminoglycan (GAG) based hydrogels were assembled in a bilayer to mimic healthy or diseased compositions of the native fibrosa and spongiosa. Multiphoton imaging and proteomic analysis of healthy and diseased VOCs were performed. RESULTS: Collagen-based bilayered hydrogel maintained the phenotype of the VICs. Proteins related to cellular processes like cell cycle progression, cholesterol biosynthesis, and protein homeostasis were found to be significantly altered and correlated with changes in cell metabolism in diseased VOCs. This study suggested that diseased VOCs may represent an early, adaptive disease initiation stage, which was corroborated by human aortic valve proteomic assessment. CONCLUSIONS: In this study, we developed a collagen-based bilayered hydrogel to mimic healthy or diseased compositions of the native fibrosa and spongiosa layers. When the gels were assembled in a VOC with VECs and VICs, the diseased VOCs revealed key insights about the CAVD initiation process. STATEMENT OF SIGNIFICANCE: Calcific aortic valve disease (CAVD) elevates the risk of death due to cardiovascular pathophysiology by 50 %, however, prevention and mitigation strategies are lacking, clinically. Developing tools to assess early disease would significantly aid in the prevention of disease and in the development of therapeutics. Previously, studies have utilized collagen and glycosaminoglycan-based hydrogels for valve cell co-cultures, valve cell co-cultures in dynamic environments, and inorganic polymer-based multilayered hydrogels; however, these approaches have not been combined to make a physiologically relevant model for CAVD studies. We fabricated a bi-layered hydrogel that closely mimics the aortic valve and used it for valve cell co-culture in a dynamic platform to gain mechanistic insights into the CAVD initiation process using proteomic and multiphoton imaging assessment.

miRNAs and NFKB1 and TRAF6 target genes: The initial functional study in CD14+ monocytes in rheumatoid arthritis patients.

We predicted miRNAs with regulatory impact on NFKB1 and TRAF6 gene expression and selected the miR-194-5p, miR-124-3p, miR-9-5p, and miR-340-5p and their target genes for expression analyses on CD14+ monocytes from rheumatoid arthritis (RA) patients and healthy controls. Additionally, we evaluated the influence of genes and miRNA expression on RA patients' cytokine levels. No difference was observed in genes or miRNAs expression when compared to healthy controls and RA patients or clinical parameters. However, we found a significant difference between miR-194-5p and miR-9-5p levels (FC=-2.31; p=0.031; FC=-3.05;p=0.031, respectively) and non-prednisone users as compared to prednisone using patients. We conducted correlation analyses to identify the strength of the relationship between expression data and cytokine plasma levels. We observed a moderate positive correlation between miR-124-3p expression and IL-6 plasma levels (r=0.46; p=0.033). In addition, overexpression of miRNAs was concomitant to TRAF6 and NFKB1 genes as indicated by correlation analyses: TRAF6 and miR-194-5p (r=0.60;p<0.001) and miR-9-5p (r=0.63;p<0.001) and NFKB1 and miR-194-5p (r=0.72;p<0.001), miR-9-5p (r=0.72;p<0.001) and miR-340-5p (r=0.61;p<0.001). NFKB1 and TRAF6 genes and miRNAs monocyte expression do not appear to be related to RA but showed a significant difference in different groups of RA therapy. In addition, increased levels of miRNAs can be linked to concomitant overexpression of TRAF6 and NFKB1 in monocytes and act as its regulators.

Expression of the miR-9-5p, miR-125b-5p and its target gene NFKB1 and TRAF6 in childhood-onset systemic lupus erythematosus (cSLE).

Childhood- onset systemic lupus erythematosus (cSLE) is a multisystem inflammatory disease that can lead to severe clinical conditions resulting in early comorbidities. Several genetic, environmental, and immunological factors are known to influence the onset of the disease. MiRNAs have been already considered as potential actors involved in the development and activity of the SLE. Thus, understanding the behavior of these regulators can contribute to clarify the inflammatory process affecting SLE patients. Among miRNAs, miR-125b-5p and miR-9-5p targeting NFKB1 and TRAF6 genes can be involved in the etio-pathogenesis of the disease by modulating inflammation. In this study we evaluated miR-9-5p and miR-125b-5p expression and its target genes NFKB1 and TRAF6 in peripheral blood samples (PBMC) from the 35 cSLE patients and 35 healthy controls. MiRNAs and gene target expression have been evaluated by using RT-PCR with specific TaqMan® probes. Both miR-9-5p [Fold Change (FC) = -2.21; p = 0.002] and miR-125b-5p (FC= -3.30; p < 0.0001) and NFKB1 (FC = -1.84; p < 0.001) were downregulated in cSLE patients, while TRAF6 was upregulated (FC = 1.80; p = 0.006) in cSLE patients when compared to controls. A significant correlation was found between miR-125b-5p and its target gene NFKB1 [Spearman (r) = 0.47; p = 0.023]. Our results showed miR-125b-5p and miR-9-5p differential expression in cSLE patients, possibly contributing to better understanding the role of these regulators in cSLE development and disease pathogenesis.

Collection of polycyclic aromatic sulfur heterocycles from asphalt fumes and quantification by an HPLC-DAD method.

A chromatographic method is described for the separation and quantification of polycyclic aromatic sulfur heterocycles (PASHs) using liquid chromatography coupled with diode array detection (DAD). The PASHs that were investigated in this study were chosen based on their similarity in molecular weight, volatility and polarity of their PAH analogues. The chromatographic separation of the compounds was optimized, and their analytical characteristics were evaluated. The limits of detection and quantification ranged from 0.05 for 2,3,4,7-tetramethylbenzothiophene to 2.16 µg L-1 for thieno[2,3-b]thiophene and from 0.16 for 2,3,4,7-tetramethylbenzothiophene to 6.53 µg L-1 for thieno[2,3-b]thiophene, respectively. Recoveries ranged from 84.9 for benzo[b]benzo[4,5]thieno[2,3-d]thiophene to 110% for dinaphtho[2,1-b:1',2'-d]thiophene. Intermediate precisions and repeatabilities lie between 1.4 and 3.0%, and 0.3 and 1.7%, respectively. The chromatographic method was applied for determination of PASHs directly in asphalt fumes, which were obtained by heating asphalt samples in a homemade closed system. The mutual interference of PAH analogues was also discussed. The method was successfully used for PASH determination in asphalt samples obtained from three different oil refineries in Brazil.

CIITA gene polymorphism (rs3087456) in systemic lupus erythematosus and rheumatoid arthritis: A population-based cohort study.

Systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA) are influenced by genetic variants in immune system HLA genes. The Class II Major Histocompatibility Complex Transactivator (CIITA) is an important co-activator of the HLA transcriptional complex; the single nucleotide variant (SNV) rs3087456 localized in the gene promoter region (-168 A/G) has been reported as able to modify its transcription level. In our study, we assessed CIITA rs3087456 SNV in 1,044 Brazilians from two Brazilian regions (Northeast and South) to verify the association with susceptibility and clinical manifestations of (SLE) and (RA) using TaqMan SNP Genotyping Assays System. We observed a protection for a recessive model (GG x AA+AG) for RA susceptibility and increased risk for erosion development in AG genotype patients. No significant association was observed for SLE susceptibility; however, we observed significant increased risk for Class IV and V nephritis development in G allele and GG genotype patients. In conclusion, we showed the contribution of CIITA rs3087456 to SLE or RA clinical features and RA susceptibility in the studied populations.

Demographic, clinical, and ancestry characterization of a large cluster of mucopolysaccharidosis IV A in the Brazilian Northeast region.

Mucopolysaccharidosis (MPS) IVA is a rare autosomal recessive disease with a highly variable distribution worldwide. Discrepancies in the incidence of MPS IVA among populations of different ethnicities are mostly attributed to founder effects. Demographic and clinical data from 28 MPS IVA patients, followed at a single center, and ancestry (Y chromosome and mitochondrial markers) of a subsample of 17 patients, most with the p.Ser341Arg (c.1023C>G) mutation were analyzed. Parental consanguinity was observed in 15/20 couples; a rare homozygous N-acetylgalactosamine-6-sulfatase (GALNS) mutation was found in 7/16 families with intra-familial phenotypic heterogeneity. Paternal ancestry was 94.2% (16/17) European, 5.8% (1/17) African, and 0% Amerindian. The European paternal haplogroups R1a, R1b, and R* accounted for 94.2% (16/17) of the patients. The R1b haplogroup, identified in 59% (10/17) of the patients, is frequently found in populations from the Iberian Peninsula. European, Amerindian, and African maternal ancestry was observed in 46.9% (8/17), 35.4% (6/17), and 17.7% (3/17) of the patients, respectively. Study of a cluster of MPS IVA patients from Northeastern Brazil, with high parental consanguinity and phenotypic heterogeneity showed predominantly European parental ancestry. This ancestry finding corroborates historical data on the local settlement, formed predominantly by European men.

The epigenetics of the hypothalamic-pituitary-adrenal axis in fetal development.

The hypothalamic-pituitary-adrenal (HPA) axis is an important hormonal mechanism of the human body and is extremely programmable during embryonic and fetal development. Analyzing its development in this period is the key to understanding in fact how vulnerabilities of congenital diseases occur and any other changes in the phenotypic and histophysiological aspects of the fetus. The environment in which the mother is exposed during the gestational period can influence this axis. Knowing this, our objective was to analyze in recent research the possible impact of epigenetic programming on the HPA axis and its consequences for fetal development. This review brought together articles from two databases: ScienceDirect and PUBMED researched based on key words such as "epigenetics, HPA axis, cardiovascular disease, and circulatory problems" where it demonstrated full relevance in experimental and scientific settings. A total of 101 articles were selected following the criteria established by the researchers. Thus, it was possible to verify that the development of the HPA axis is directly related to changes that occur in the cardiovascular system, to the cerebral growth and other systems depending on the influence that it receives in the period of fetal formation.

Equilin displays similar endothelium-independent vasodilator potential to 17ß-estradiol regardless of lower potential to inhibit calcium entry.

Conjugated equine estrogens (CEE) have been widely used by women who seek to relieve symptoms of menopause. Despite evidence describing protective effects against risk factors for cardiovascular diseases by naturally occurring estrogens, little is known about the vascular effects of equilin, one of the main components of CEE and not physiologically present in women. In this regard, the present study aims to compare the vascular effects of equilin in an experimental model of hypertension with those induced by 17ß-estradiol. Resistance mesenteric arteries from female spontaneously hypertensive rats (SHR) were used for recording isometric tension in a small vessel myograph. As effectively as 17ß-estradiol, equilin evoked a concentration-dependent relaxation in mesenteric arteries from female SHRs contracted with KCl, U46619, PDBu or ET-1. Equilin-induced vasodilation does not involve classical estrogen receptor activation, since the estrogen receptor antagonist (ICI 182,780) failed to inhibit relaxation in U46619-precontracted mesenteric arteries. Vasorelaxation was not affected by either endothelium removal or by inhibiting the release or action of endothelium-derived factors. Incubation with L-NAME (NOS inhibitor), ODQ (guanylyl cyclase inhibitor) or KT5823 (inhibitor of protein kinase G) did not affect equilin-induced relaxation. Similarly, indomethacin (COX inhibitor) or blockage of potassium channels with tetraethylammonium, glibenclamide, 4-aminopyridine, or ouabain did not affect equilin-induced relaxation. Inhibitors of adenylyl cyclase SQ22536 or protein kinase A (KT5720) also had no effects on equilin-induced relaxation. While 17ß-estradiol inhibited calcium (Ca2+) -induced contractions in high-K+ depolarization medium in a concentration-dependent manner, equilin induced a slight rightward-shift in the contractile responses to Ca2+. Comparable pattern of responses were observed in the concentration-response curves to (S)-(-)-Bay K 8644, a L-type Ca2+ channel activator. Equilin was unable to block the transitory contraction produced by caffeine-induced Ca2+ release from intracellular stores. In conclusion, equilin blocks L-type Ca2+ channels less effectively than 17ß-estradiol. Despite its lower effectiveness, equilin equally relaxes resistance mesenteric arteries by blocking Ca2+ entry on smooth muscle.

Efeito da contaminação por saliva na resistência adesiva de braquetes cerâmicos utilizando uma resina ortodôntica hidrofílica / Saliva contamination effect on shear bond strength of ceramic brackets with an orthodontic hydrophilic resin

Introdução: Alguns fatores, como a presença de saliva, são capazes de influenciar a adesão do braquete ao dente durante o procedimento de colagem e podem causar falha da resistência adesiva. Objetivo: O objetivo deste estudo foi avaliar a resistência ao cisalhamento de braquetes cerâmicos ortodônticos colados com Transbond XT e Transbond Plus Color Change em esmalte de dentes bovinos, contaminado e não contaminado por saliva, além de analisar o local da falha adesiva. Material e método: Sessenta incisivos bovinos foram divididos aleatoriamente em quatro grupos (n=15), de acordo com o material de colagem e a presença de contaminação: Grupo 1 (G1): colagem com Transbond XT na ausência de contaminação; Grupo 2 (G2): colagem com Self Etching Pimer e Transbond Color Change na ausência de contaminação; Grupo 3 (G3): colagem com Transbond XT na presença de contaminação, e Grupo 4 (G4): colagem com Self Etching Primer e Transbond Color Change na presença de contaminação. Resultado: O teste não paramétrico de Kruskal-Wallis mostrou que G1 diferiu estatisticamente (p<0,05) de G2 e G3. Não houve diferença estatística significativa entre os demais grupos. O índice de adesivo remanescente (IAR) variou entre 2 e 3 no G1 e entre 0 e 1 nos outros grupos. Conclusão: A contaminação por saliva diminui a resistência adesiva ao cisalhamento de braquetes cerâmicos colados com a resina hidrofóbica Transbond XT convencional. Por outro lado, a utilização da resina hidrofílica Transbond Plus Color Change associada ao Self Etching Primer, em ambiente contaminado por saliva, confere resistência adesiva adequada para o seu uso clínico.

Introduction: Some fator, such as the presence of saliva, are able to influence the adhesion of the bracket to the tooth enamel during the orthodontic bonding procedure and may cause the adhesive failure. Objective: The aim of this study was to evaluate the shear strength of orthodontic ceramic brackets bonded with Transbond XT and Transbond Plus Color Change to bovine teeth, under saliva contamination and the adhesive failure mode. Material and method: Sixty bovine permanent incisors were randomly divided into 4 groups (n=15) according to bonding material and saliva contamination: Group 1 (G1) bonded with Transbond XT with no saliva contamination; Group 2 (G2) bonded with Self Etching Primer and Transbond Plus Color Change with no saliva contamination; Group 3 (G3) bonded with Transbond XT with saliva contamination and Group 4 (G4) bonded with Transbond Plus Color Change with saliva contamination. Result: The Kruskal-Wallis non-parametric test (p<0,05) was used and showed that G1 differed statistically from G2 and G3. There was no statistical difference between the other groups. The adhesive remnant index (ARI) ranged between 2 and 3 in G1 and between 0 and 1 score in the other groups. Conclusion: Saliva contamination decreases the shear bond strength of ceramic brackets bonded with conventional Transbond XT hydrophobic resin. On the other hand, the Transbond Plus Color Change hydrophilic resin associated with a Self Etching Primer, under saliva contamination, provide a sufficient shear bond strength, suitable for their clinical use.

Benefits of intradialytic physiotherapy in quality of life, pain, edema and respiratory function of patients with chronic kidney disease / Benefícios da fisioterapia intradialítica na qualidade de vida, dor, edema e função respiratória de doentes renais crônicos / Beneficios de la fisioterapia intradialítica en la calidad de la vida, el dolor, el edema y la función respiratoria de los pacientes renales crónicos

Abstract Introduction: Patients with chronic kidney disease suffer from a decline in quality of life and respiratory function, for various reasons related to this condition. Objective: To verify the influence of intradialytic physiotherapy on the quality of life and respiratory function in chronic renal patients. Methods: The sample was non-probabilistic and consisted of 51 individuals, of both sexes, aged between 30 and 60 years. The WHOQOL-Bref, an evaluation form, the Manovacuometer and Peak Flow were used to detect the impact of the physiotherapeutic intervention before and after an eight-week treatment protocol. Results: There was a difference in the comparison of the general QOL before and after (p = 0.006) the intervention, as well as in the improvement of the maximal forced expiratory flow peak (p = 0.001), the PEmax (p = 0.000), peak forced expiratory flow rate (0.017) and pain (p = 0.006). There was also improvement of edema (p = 0.013) and cramps (p = 0.000). Conclusion: The benefits of intradialytic physiotherapy in improving QOL and respiratory function in chronic kidney patients are explicit.

Resumo Introdução: Pacientes com doença renal crônica sofrem declínio na qualidade de vida e função respiratória, por diversos motivos ligados à afecção. Objetivo: Verificar a influência da fisioterapia intradialítica sobre a qualidade de vida e a função respiratória em pacientes renais crônicos. Métodos: A amostra foi não probabilística e composta por 51 indivíduos, de ambos os sexos, com idade estabelecida entre 30 e 60 anos. Utilizaram-se o WHOQOL-Bref, uma ficha de avaliação, o Manovacuômetro e o Pico de fluxo expiratório forçado máximo (Peak Flow) para detectar o impacto da intervenção fisioterapêutica antes e após um protocolo de oito semanas de tratamento. Resultados: Houve diferença na comparação da QV geral antes e depois (p =0,006) da intervenção, assim como na melhora da PImáx (p = 0,001), da PEmáx (p = 0,000), do pico de fluxo expiratório forçado máximo (p = 0,017) e da EVA de dor (p = 0,006). Também houve melhora do edema (p = 0,013) e das câimbras (p = 0,000). Conclusão: Ficam explícitos os benefícios da fisioterapia intradialítica na melhora da QV e a função respiratória de doentes renais crônicos.

Resumen Introducción: Los pacientes con enfermedad renal crónica sufren una disminución de la calidad de vida y la función respiratoria, por diversos motivos relacionados con la afección. Objetivo: Verificar la influencia de la Fisioterapia intradialítica sobre la calidad de vida, función respiratoria en pacientes renales crónicos. Métodos: La muestra fue no probabilística y compuesta por 51 individuos, de ambos sexos, con edad establecida entre 30 a 60 años. Se utilizó el WHOQOL-Bref, una ficha de evaluación, el Manovacuómetro y el Pico de flujo espiratorio forzado máximo (Peak Flow) para detectar el impacto de la intervención fisioterapéutica antes y después de un protocolo de ocho semanas de tratamiento. (P = 0,006) de la intervención, así como en la mejora de la PImáx (p = 0,001), de la PEmáx (p = 0,000), del pico de flujo espiratorio forzado máximo (p = 0,000), en la comparación de la QV general antes y después (p = 0,006) 0,017) y de la EVA de dolor (p = 0,006). También hubo una mejora del edema (p = 0,013) y de los calambres (p = 0,000). Conclusión: Queda explícito los beneficios de la fisioterapia intradialítica en la mejora de la QV y la función respiratoria de pacientes renales crónicos.

Methylmercury exposure for 14 days (short-term) produces behavioral and biochemical changes in mouse cerebellum, liver, and serum.

Various studies on methylmercury (MeHg)-induced toxicity focused on the central nervous system (CNS) as a primary target. However, MeHg-mediated toxicity is related to metallic interaction with electrophilic groups, which are not solely restricted to the CNS, but these reactive groups are present ubiquitously in several systems/organs. The aim of this study was thus to examine MeHg-induced systemic toxicity in mice using a standardized neurotoxicology testing exposure model to measure cerebellar neurotoxicity by determining biochemical and behavioral parameters in the cerebellum. After 2 weeks exposure to MeHg (40 µg/ml; diluted in drinking water; ad libitum), adult male Swiss mice showed a marked motor impairment characteristic of cerebellar toxicity as noted in the following tests: rotarod, beam walking, pole, and hind limb clasping. MeHg treatment resulted in Hg deposition in the cerebellum as well as reduction in cerebellar weight, glutathione peroxidase (GPx) activity, and interleukin (IL)-6 levels. MeHg ingestion increased cerebellar glutathione reductase (GR) activity and brain-derived neurotrophic factor (BDNF) levels. In addition to cerebellar toxicity, MeHg treatment also elevated total and non-high density lipoprotein (non-HDL) cholesterol levels, as well as serum aspartate transaminase (AST) and alanine transaminase (ALT) enzymatic activities, systemic parameters. Increased liver weight and reduced serum urea levels were also noted in MeHg-exposed mice. Taken together, our findings demonstrated that a well-standardized exposure protocol to examine MeHg-induced neurotoxicity also produced systemic toxicity in mice, which was characterized by changes in markers of hepatic function as well as serum lipid homeostasis.

Serosurvey of Borrelia in dogs, horses, and humans exposed to ticks in a rural settlement of southern Brazil.

The aims of the present study were to serosurvey dogs, horses, and humans highly exposed to tick bites for anti-Borrelia burgdorferi s.l. antibodies, identify tick species present, and determine risk factors associated with seropositivity in a rural settlement of Paraná State, southern Brazil. Eighty-seven residents were sampled, along with their 83 dogs and 18 horses, and individual questionnaires were administered. Immunofluorescence antibody test (IFAT) was performed on serum samples and positive samples were subjected to western blot (WB) analysis. Anti-B. burgdorferi antibodies were found in 4/87 (4.6%) humans, 26/83 (31.3%) dogs, and 7/18 (38.9%) horses by IFAT, with 4/4 humans also positive by WB. Ticks identified were mostly from dogs and included 45/67 Rhipicephalus sanguineus, 21/67 Amblyomma ovale, and 1/67 A. cajennense sensu lato. All (34/34) horse ticks were identified as A. cajennense s.l.. No significant association was found when age, gender, or presence of ticks was correlated to seropositivity to Borrelia sp. In conclusion, although anti-Borrelia antibodies have been found in dogs, horses and their owners from the rural settlement, the lack of isolation, molecular characterization, absence of competent vectors and the low specificity of the commercial WB kit used herein may have impaired risk factor analysis.

Efeito do NaOCl na resistência de união de pinos de fibra cimentados à dentina utilizando um cimento convencional e um autoadesivo / Effects of NaOCl on bond strength of fiber posts cemented to dentin using a conventional and a self-adhesive cement

Objetivo: avaliar o efeito do hipoclorito de sódio (NaOCl) na resistência de união (RU) de pinos de fibra de vidro cimentados à dentina, utilizando o cimento resinoso convencional AllCem ou o autoadesivo RelyX U200. Material e Métodos: foram utilizados 40 dentes bovinos, que tiveram suas coroas removidas e as raízes padronizadas em 7mm, referente aos terços cervical e médio. Os condutos foram preparados e as raízes divididas em 4 grupos (n = 10). Nos grupos AD/U200 e AD/AAC os condutos foram irrigados com água destilada. Nos grupos NaOCl/U200 e NaOCl/AAC, os condutos foram imersos em NaOCl a 5,25% durante 20 minutos e, em seguida, irrigados com água destilada. Os pinos foram cimentados ao conduto com o cimento RelyX U200 nos grupos AD/U200 e NaOCl/U200, e com o cimento Allcem nos grupo AD/AAC e NaOCl/AAC. As raízes foram segmentadas e os discos submetidos a ensaio de push-out. Os valores de RU foram analisados estatisticamente pelos testes de Kruskal-Wallis e Mann Whitney (α = 0,05). A análise do padrão de fratura das amostras foi realizada em lupa estereoscópica. Resultados: o NaOCl mostrou um efeito deletério na resistência adesiva para ambos os cimentos. Os valores de RU foram superiores para os grupos Âmbar/AllCem em comparação aos grupos RelyX U200. Em relação ao padrão de fratura, houve predomínio de fratura tipo I, adesiva cimento/dentina. Conclusão: o NaOCl mostrou um efeito deletério na resistência de união de pinos de fibra cimentados à dentina utilizando os cimentos RelyX U200 e Âmbar/AllCem.

Objective: to evaluate the effect of sodium hypochlorite (NaOCl) on the bond strength (BS) of fiberglass posts cemented to dentin using the conventional cement Allcem or the self-adhesive cement RelyX U200. Materials and Methods: forty bovine teeth had their crowns removed and the roots standardized in 7 mm, referring to the cervical and middle thirds. The root canals were prepared and roots were divided into four groups (n = 10). In AD/U200 and AD/AAC groups, the root canals were irrigated with distilled water. In NaOCl/U200 and NaOCl/AAC groups, the root canals were immersed in 5.25% NaOCl for 20 min and then irrigated with distilled water. The posts were cemented with RelyX U200 in groups AD/U200 and NaOCl/U200, and with Allcem cement in groups AD/AAC and NaOCl/AAC. The roots were segmented and discs were subjected to the push-out test. The BS values were statistically analyzed by Kruskal-Wallis and Mann-Whitney tests (α = 0.05). The fracture pattern was analyzed using a stereomicroscope. Results: the NaOCl showed a deleterious effect on bond strength for both cements. The RU values were higher for Ambar/Allcem groups compared with the U200 groups. Regarding the fracture pattern, there was a predominance of type I, adhesive cement/dentin. Conclusion: NaOCl showed a deleterious effect on the bond strength of fiber posts cemented to dentin using RelyX U200 and Allcem cements.

Serosurvey of Borrelia in dogs, horses, and humans exposed to ticks in a rural settlement of southern Brazil / Soroprevalência e fatores associados a Borrelia em cães, equinos e humanos expostos a carrapatos em um assentamento rural do sul do Brasil

Abstract The aims of the present study were to serosurvey dogs, horses, and humans highly exposed to tick bites for anti-Borrelia burgdorferi s.l. antibodies, identify tick species present, and determine risk factors associated with seropositivity in a rural settlement of Paraná State, southern Brazil. Eighty-seven residents were sampled, along with their 83 dogs and 18 horses, and individual questionnaires were administered. Immunofluorescence antibody test (IFAT) was performed on serum samples and positive samples were subjected to western blot (WB) analysis. Anti-B. burgdorferi antibodies were found in 4/87 (4.6%) humans, 26/83 (31.3%) dogs, and 7/18 (38.9%) horses by IFAT, with 4/4 humans also positive by WB. Ticks identified were mostly from dogs and included 45/67 Rhipicephalus sanguineus, 21/67 Amblyomma ovale, and 1/67 A. cajennense sensu lato. All (34/34) horse ticks were identified as A. cajennense s.l.. No significant association was found when age, gender, or presence of ticks was correlated to seropositivity to Borrelia sp. In conclusion, although anti-Borrelia antibodies have been found in dogs, horses and their owners from the rural settlement, the lack of isolation, molecular characterization, absence of competent vectors and the low specificity of the commercial WB kit used herein may have impaired risk factor analysis.

Resumo Os objetivos do presente estudo foram realizar um levantamento sorológico de cães, cavalos e humanos altamente expostos a picadas de carrapatos para anticorpos anti-B. burgdorferi s.l., identificar as espécies de carrapatos presentes, e determinar os fatores de risco associados a soropositividade em um assentamento rural do Estado do Paraná, sul do Brasil. Oitenta e sete residentes foram amostrados junto com seus respectivos 83 cães e 118 cavalos e questionários individuais foram aplicados. O teste de imunofluorescência indireta (IFI) foi realizado nas amostras sorológicas e as positivas foram submetidas a análise por western blot (WB). Anticorpos anti-B. burgdorferi foram detectados em 4/87 (4,6%) humanos, 26/83 (31,3%) cães e 7/18 (38,9%) cavalos pela IFI, com 4/4 humanos também positivos pelo WB. Os carrapatos identificados foram em sua maioria de cães e incluíram 45/67 Rhipicephalus sanguineus, 21/67 Amblyomma ovale e 1/67 A. cajennense sensu lato. Todos (34/34) carrapatos dos cavalos foram identificados como A. cajennense s.l.. Não foram observadas diferenças estatísticas entre idade, sexo ou presença de carrapatos e soropositividade para Borrelia sp. Em conclusão, embora anticorpos anti-Borrelia tenham sido encontrados em cães, equinos e seus proprietários do assentamento rural, a ausência de isolamento, caracterização molecular, ausência de vetores competentes e baixa especificidade do kit comercial de WB utilizado podem ter limitado a análise de fatores de risco.

Feira da Saúde - prevenção e promoção da saúde no ambiente escolar / Health Fair - prevention and health promotion in the school environment

O ambiente escolar é muito importante para desenvolver, no processo de ensino, as ações de saúde. Assim, o NASF Continental e a UBS Paulista, junto com parceiros, desenvolveu uma Feira da Saúde na Escola Estadual Roberta Maria Lopes Chaves, com o objetivo de ampliar o conceito de saúde, mobilizando para o autocuidado e autonomia. Foi realizado um circuito com dez salas temáticas: Saúde Bucal; Higiene Pessoal; Guarda Responsável; Circuito Sensorial; Grafite; Bullying; Alimentação Saudável; Lixo; Água Virtual; e Medicação. Participaram dessa atividade 682 alunos, da 1ª à 8ª série. O objetivo foi alcançado, visto que a proposta principal era agregar conhecimento sobre saúde, valorizando a participação das crianças nessa construção de saber, o que foi constatado durante as atividades com perguntas e grande interação das mesmas.

Serosurvey of tick-borne pathogens in dogs from urban and rural areas from Parana State, Brazil.

Considering the zoonotic potential of tick-borne disease (TBD) agents and the fact that dogs may act as sentinels for human infection, the aim of the present study was to determine the seroprevalence of TBD agents and risk factors for exposure in two different canine populations from Parana State, Southern Brazil. A total of 138 dog serum samples from urban (UA) (n=68) and rural (RA) (n=70) areas were tested with commercial ELISA rapid test for Anaplasma phagocytophilum, Ehrlichia canis and Borrelia burgdorferi antibodies and indirect immunofluorescence assay (IFAT) for Babesia vogeli. An overall of 92∕138 (66.7%) dogs, being 62∕68 (91.2%) from UA and 30∕70 (42.9%) from RA, were seropositive for at least one TBD agent. From the total number of dogs, sixty-two were positive for E. canis (44.9%), 19 (13.8%) for A. phagocytophilum, and 64 (46.4%) for B. vogeli. Anti-B. burgdorferi antibodies were not detected. Dogs from UA showed a higher percentage of tick infestation (p = 0.0135) and were highly associated with seropositivity to E. canis (p = 0.000005), A. phagocytophilum (p = 0.0001), and B. vogeli (p = 0.0012). In summary, the findings indicate that dogs from urban areas present higher potential risk exposure to TBD pathogens than those from rural areas.

Seroepidemiological survey of Theileria equi and Babesia caballi in horses from a rural and from urban areas of Paraná State, southern Brazil.

The objective of this study was to evaluate the seroepidemiological data of Babesia caballi and Theileria equi in horses from a rural settlement and carthorses from urban areas of Paraná State, southern Brazil. A total of 198 horses, including 32 from the rural settlement and 166 carthorses from Colombo (n=48), Pinhais (n=76), Londrina (n=24), and Curitiba city (n=18) was sampled and tested using a commercial competitive inhibition ELISA (cELISA) test. Out of the 198 horses, 193 (97.5%) were seropositive for at least one piroplasm species. Antibodies to T. equi were detected in 155/198 horses (78.3%), antibodies to B. caballi were detected in 137/198 horses (69.2%), and antibodies to both were detected in 99/198 (50.0%) horses. Horses living in the rural settlement and Colombo were more likely to be seropositive to T. equi than those in Curitiba (p<0.05). Horses older than 5 years were more likely to be seropositive for T. equi than those younger than 5 years (p<0.05). No significant association was found between gender or the presence of ticks and seropositivity to T. equi (p>0.05). In conclusion, the high seroprevalences to B. caballi and T. equi observed in this study emphasize that active surveillance programs are critical for monitoring animal health status, particularly because carthorses may act as urban disseminators of these piroplasms.

Serological survey of Ehrlichia species in dogs, horses and humans: zoonotic scenery in a rural settlement from southern Brazil.

The aims of this study were to determine the seroprevalence of Ehrlichia spp. and risk factors for exposure in a restricted population of dogs, horses, and humans highly exposed to tick bites in a Brazilian rural settlement using a commercial ELISA rapid test and two indirect immunofluorescent assays (IFA) with E. canis and E. chaffeensis crude antigens. Serum samples from 132 dogs, 16 horses and 100 humans were used. Fifty-six out of 132 (42.4%) dogs were seropositive for E. canis. Dogs > one year were more likely to be seropositive for E. canis than dogs ≤ one year (p = 0.0051). Ten/16 (62.5%) and 8/16 (50%) horses were seropositive by the commercial ELISA and IFA, respectively. Five out of 100 (5%) humans were seropositive for E. canis and E. chaffeensis. Rhipicephalus sanguineus (n = 291, 97.98%) on dogs and Amblyomma cajennense (n = 25, 96.15%) on horses were the most common ticks found. In conclusion, anti-Ehrlichia spp. antibodies were found in horses; however, the lack of a molecular characterization precludes any conclusion regarding the agent involved. Additionally, the higher seroprevalence of E. canis in dogs and the evidence of anti-Ehrlichia spp. antibodies in humans suggest that human cases of ehrlichiosis in Brazil might be caused by E. canis, or other closely related species.

SEROLOGICAL SURVEY OF Ehrlichia SPECIES IN DOGS, HORSES AND HUMANS: ZOONOTIC SCENERY IN A RURAL SETTLEMENT FROM SOUTHERN BRAZIL / Investigação sorológica de espécies de Ehrlichia em cães, equinos e humanos de um assentamento rural do sul do Brasil

SUMMARY The aims of this study were to determine the seroprevalence of Ehrlichia spp. and risk factors for exposure in a restricted population of dogs, horses, and humans highly exposed to tick bites in a Brazilian rural settlement using a commercial ELISA rapid test and two indirect immunofluorescent assays (IFA) with E. canis and E. chaffeensis crude antigens. Serum samples from 132 dogs, 16 horses and 100 humans were used. Fifty-six out of 132 (42.4%) dogs were seropositive for E. canis. Dogs > one year were more likely to be seropositive for E. canis than dogs ≤ one year (p = 0.0051). Ten/16 (62.5%) and 8/16 (50%) horses were seropositive by the commercial ELISA and IFA, respectively. Five out of 100 (5%) humans were seropositive for E. canis and E. chaffeensis. Rhipicephalus sanguineus (n = 291, 97.98%) on dogs and Amblyomma cajennense (n = 25, 96.15%) on horses were the most common ticks found. In conclusion, anti-Ehrlichia spp. antibodies were found in horses; however, the lack of a molecular characterization precludes any conclusion regarding the agent involved. Additionally, the higher seroprevalence of E. canis in dogs and the evidence of anti-Ehrlichia spp. antibodies in humans suggest that human cases of ehrlichiosis in Brazil might be caused by E. canis, or other closely related species. .

RESUMO Objetivou-se determinar a soroprevalência de Ehrlichia spp. e os fatores de risco associados a exposição em uma população restrita de cães, cavalos e humanos altamente expostos a picadas de carrapatos em um assentamento rural brasileiro utilizando um teste comercial de ELISA rápido e dois testes de imunofluorescência indireta (IFI) com antígenos brutos de E. canis e E. chaffeensis. Amostras de soro de 132 cães, 16 cavalos e 100 humanos foram utilizadas. Cinquenta e seis/132 (42,4%) cães foram soropositivos para E. canis. Cães > um ano apresentaram mais chance de serem soropositivos para E. canis do que cães ≤ um ano (p = 0,0051). Dez/16 (62,5%) e 8/16 (50%) cavalos foram soropositivos pelo ELISA comercial e IFI, respectivamente. Cinco/100 (5%) humanos foram soropositivos para E. canis e E. chaffeensis. Rhipicephalus sanguineus (n = 291, 97,98%) nos cães e A. cajennense (n = 25, 96,15%) nos cavalos foram os carrapatos mais encontrados. Concluindo, anticorpos anti-Ehrlichia spp. foram encontrados em cavalos; entretanto, a ausência de uma caracterização molecular impede qualquer conclusão sobre agente envolvido. Além disso, a alta soroprevalência de E. canis em cães e a evidência de anticorpos anti-Ehrlichia sp. em humanos, sugere que os casos de erliquiose humana no Brasil possam ser causados por E. canis ou outra espécie intimamente relacionada. .

Immune response of calves inoculated with proteins of Anaplasma marginale bound to an immunostimulant complex.

Despite our current knowledge of the immunology, pathology, and genetics of Anaplasma marginale, prevention in cattle is currently based on old standbys, including live attenuated vaccines, antibiotic treatment, and maintaining enzootic stability in cattle herds. In the present study, we evaluated the use of an immunostimulant complex (ISCOMATRIX) adjuvant, associated with a pool of recombinant major surface proteins (rMSP1a, rMSP1b, rMSP4 and rMSP5) to improve the humoral immune response triggered in calves mainly by IgG2. Ten calves were divided in three groups: 4 calves were inoculated with the ISCOMATRIX/rMSPs (G1); 2 calves were inoculated with ISCOMATRIX adjuvant (G2); and 4 calves received saline (G3). Three inoculations were administered at 21-day intervals. In G1, the calves showed significant increases in total IgG, IgG1 and IgG2 levels 21 days after the second inoculation, compared to the control group (p < 0.05), and G1 calves remained above the cut-off value 28 days after the third inoculation (p < 0.05). The post-immunized sera from calves in G1 reacted specifically for each of the rMSPs used. In conclusion, the ISCOMATRIX/rMSPs induced antigen-specific seroconversion in calves. Therefore, additional testing to explore the protection induced by rMSPs, both alone and in conjunction with proteins previously identified as subdominant epitopes, is warranted.