Abstract The genus Neorickettsia comprises trematode-associated bacteria that can cause diseases in animals and humans. Despite detection of Neorickettsia antigens in the intestine of coatis kept in captivity in southern Brazil through immunohistochemistry, the molecular identity of the bacteria in South American procyonids remains elusive. The aim of the present study was to investigate the occurrence of Neorickettsia sp. in blood samples from coatis in central-western Brazil. Between March 2018 and January 2019, animals were captured and recaptured in two areas of the Cerrado (Parque Estadual do Prosa, PEP; and Vila da Base Aérea, VBA) located in the city of Campo Grande, state of Mato Grosso do Sul, central-western Brazil. All captures were performed according to convenience. DNA from 97 blood samples was subjected to nested PCR (nPCR) targeting a fragment of the 16S rRNA gene of Neorickettsia sp. Six samples (3.6%; five from VBA and one from PEP) from different coatis were positive in nPCR based on the 16S rRNA. The sequences obtained (~500 bp) showed ˃ 99% similarity to N. risticii. Phylogenetic analysis clustered the sequences detected in the present study in a clade with N. risticii. This is the first molecular detection of Neorickettsia sp. in coatis in Brazil. trematodeassociated trematode associated humans immunohistochemistry elusive sp centralwestern central western 201 2019 Parque Prosa PEP Aérea Grande Sul convenience 9 (nPCR S 3.6% 36 3 6 (3.6% ~500 500 (~50 bp 99 N risticii 20 3.6 (3.6 ~50 50 (~5 2 3. (3. ~5 5 (~ (3 ~ (

Resumo O gênero Neorickettsia compreende bactérias associadas a trematódeos que podem causar doenças em animais e humanos. Apesar da detecção de antígenos de Neorickettsia por imuno-histoquímica no intestino de quatis mantidos em cativeiro no sul do Brasil, a identidade molecular da bactéria em procionídeos da América do Sul permanece indefinida. O presente trabalho teve como objetivo investigar a ocorrência de Neorickettsia sp. em amostras de sangue de quatis do Centro-Oeste do Brasil. Entre março de 2018 e janeiro de 2019, os animais foram capturados em duas áreas de Cerrado (Parque Estadual do Prosa PEP e Vila da Base Aérea VBA) localizadas na cidade de Campo Grande, Mato Grosso do Sul, Centro-Oeste do Brasil. Todas as capturas e recapturas foram realizadas por conveniência. O DNA de 97 amostras de sangue foi submetido a "nested" (nPCR), baseada em um fragmento do gene 16S rRNA de Neorickettsia sp. Seis (3,6% - 5 de VBA e 1 de PEP) amostras de quatis diferentes foram positivas na nPCR, baseada no rRNA 16S. As sequências obtidas (~500 pb) apresentaram ˃99% de identidade com N. risticii. A inferência filogenética agrupou as sequencias detectadas no presente estudo em um clado com N. risticii. Esta é a primeira detecção molecular de Neorickettsia sp. em quatis do Brasil. humanos imunohistoquímica imuno histoquímica Brasil indefinida sp CentroOeste Centro Oeste 201 2019 Parque Grande conveniência 9 nested "nested nPCR , (nPCR) S 3,6% 36 3 6 (3,6 ~500 500 (~50 pb 99 ˃99 N risticii 20 (nPCR 3,6 (3, ~50 50 (~5 ˃9 2 3, (3 ~5 (~ ˃ ( ~

Abstract The TP53 3’UTR variant rs78378222 A>C has been detected in different tumor types as a somatic alteration that reduces p53 expression through modification of polyadenylation and miRNA regulation. Its prevalence is not yet known in all tumors. Herein, we examine tumor tissue prevalence of rs7837822 in Brazilian cohorts of patients from south and southeast regions diagnosed with lung adenocarcinoma (LUAD, n=586), sarcoma (SARC, n=188) and uterine leiomyoma (ULM, n=41). The minor allele (C) was identified in heterozygosity in 6/586 LUAD tumors (prevalence = 1.02 %) and none of the SARC and ULM samples. Additionally, next generation sequencing analysis revealed that all variant-positive tumors (n=4) with sample availability had additional pathogenic or likely pathogenic somatic variants in the TP53 coding regions. Among them, 3/4 (75 %) had the same pathogenic or likely pathogenic sequence variant (allele frequency <0.05 in tumor DNA) namely c.751A>C (p.Ile251Leu). Our results indicate a low somatic prevalence of rs78378222 in LUAD, ULM and SARC tumors from Brazilian patients, which suggests that no further analysis of this variant in the specific studied regions of Brazil is warranted. However, these findings should not exclude tumor molecular testing of this TP53 3’UTR functional variant for different populations. TP TP5 3UTR UTR 3 rs AC A C p p5 regulation Herein rs783782 (LUAD n=586, n586 n n=586 , 586 n=586) SARC, (SARC n=188 n188 188 ULM, (ULM n=41. n41 n=41 . 41 n=41) (C 6586 6 6/58 102 1 02 1.0 % samples Additionally variantpositive positive n=4 n4 4 (n=4 them 34 3/ 75 (7 005 0 05 <0.0 DNA c751AC cAC c 751A p.Ile251Leu. pIle251Leu pIleLeu p.Ile251Leu Ile251Leu Ile Leu (p.Ile251Leu) warranted However populations rs78378 n58 n=58 58 n=18 n18 18 658 6/5 10 1. n= (n= 7 ( 00 <0. pIle IleLeu (p.Ile251Leu rs7837 n5 n=5 5 n=1 n1 65 6/ (n <0 rs783 < rs78 rs7

ABSTRACT Background: Rickettsia of the spotted fever group (SFG) has been reported in ticks and domestic animals in Campo Grande (CG), Midwest Brazil. Methods: We searched for Rickettsia in the SFG in capybaras and their ticks in an urban park in the CG. Results: The seropositivity rate was 88.2% (15/17). Although 87.7% of the capybaras sampled showed infestations with Amblyomma sculptum, A. dubitatum, and Amblyomma spp., no molecular results were detected in ticks. Conclusions: Since Rickettsia from the SFG circulates among capybaras in the urban parks of Campo Grande, this large rodent species should be monitored within the One Health Agenda.

The development of visible-light active photocatalysts is vital to harvest the solar irradiation and build efficient photoelectrochemical devices. Bismuth oxyhalide (BiOCl, BiOBr and BiOI) materials stand out due to their remarkable properties, e.g., the band gap energy can be tuned from 1.9 to 3.4 eV depends on the halide employed. However, the hydrothermal synthesis of this material class was not properly investigated and compared. Then, we evaluated the effect of the hydrothermal treatment in the BiOX’s structural, electronic, morphological and textural properties and the relationship with their photocatalytic performance. Methylene blue (MB) dye was used as probe for photocatalytic performance evaluation under ultraviolet (UV) and visible (Vis) radiation. BiOCl and BiOBr samples showed a good photoactivity under both UV and visible radiation, on other hand the BiOI samples was not active, likely due to the fast charge recombination. BiOCl and BiOBr samples exhibit a favorably interaction with MB dye, because their surfaces are negatively charged. BiOCl samples were able to promote around 40 and 80% of the MB dye degradation under visible and UV radiation, respectively. The samples were also employed as photoanode for O2 evolution, BiOI sample exhibited the best performance, due to the synergistic effect of photocatalysis and electrocatalysis.

Resumo Este estudo teve como objetivo determinar a co-soropositividade para agentes transmitidos por carrapatos, como Anaplasma marginale, Babesia bovis, Babesia bigemina, Coxiella burnetii, Anaplasma phagocytophilum, e Trypanosoma vivax em bovinos de corte do Pantanal Brasileiro. Amostras de soro foram colhidas de 400 animais (200 vacas; 200 bezerros) e submetidas a Ensaios Imunoenzimáticos Indiretos (iELISA) para detecção de anticorpos IgG anti- A. marginale, anti- B. bovis, anti- B. bigemina e anti- T. vivax, e à Reação de Imunofluorescência Indireta (RIFI) para detecção de anticorpos IgG anti -C. burnetii e anti- A. phagocytophilum. Ausência de correlação foi vista entre os animais soropositivos para C. burnetii e A. phagocytophilum com os outros agentes e correlação moderada ocorreu entre A. marginale x B. bigemina x B. bovis. Vacas foram mais soropositivas que bezerros para T. vivax e bezerros mais soropositivos que vacas para B. bovis e B. bigemina. Maior número de animais soropositivos para um único agente foi visto para T. vivax (15,2%). Vacas demonstraram maior co-soropositividade para T. vivax + A. marginale (25,5%) e bezerros para T. vivax + B. bovis + B. bigemina + A. marginale (57,5%). A alta correlação entre a soropositividade para A. marginale x B. bovis x B. bigemina é provavelmente devida à presença do vetor biológico, o carrapato Rhipicephalus microplus, nas fazendas estudadas. As vias de transmissão comuns, mediadas por dípteros hematófagos e fômites, podem explicar a alta co-soropositividade das vacas para A. marginale e T. vivax. A baixa soropositividade para C. burnetii é provavelmente devida ao tipo de sistema de criação empregado (extenso). A soropositividade para A. phagocytophilum em apenas um animal sugere a ocorrência de reação sorológica cruzada com outro agente do gênero Anaplasma.

Abstract This study investigated the seropositivity for five different tick-borne agents, namely Anaplasma marginale, Babesia bovis, Babesia bigemina, Coxiella burnetii, Anaplasma phagocytophilum, and Trypanosoma vivax in beef cattle in the Brazilian Pantanal. The serum samples collected from animals (200 cows; 200 calves) were used in indirect enzyme-linked immunosorbent assays (iELISA) to detect IgG antibodies against A. marginale, B. bovis, B. bigemina, and T. vivax, and Indirect Fluorescent Antibody Test (IFAT) for detecting IgG antibodies against C. burnetii and A. phagocytophilum. No correlation was observed between seropositivity for C. burnetii and A. phagocytophilum with other agents whereas moderate correlation was observed for A. marginalexB. bigemina x B. bovis. Cows were more seropositive for T. vivax whereas calves were more seropositive for B. bovis and B. bigemina. The highest number of seropositive animals by a single agent was observed for T. vivax (15.2%). Co-seropositivity for T. vivax + A. marginale was higher in cows (25.5%) and for T. vivax + B. bovis + B. bigemina + A. marginale was higher in calves (57.5%). The high seropositivity correlation for A. marginale x B. bovis x B. bigemina is probably due to the presence of the tick biological vector, Rhipicephalus microplus, in the studied farms. Common transmission pathways, mediated by hematophagous dipterans and fomites, may explain the high co-seropositivity of cows for A. marginale and T. vivax. Low seropositivity to C. burnetii is probably due to the type of breeding system employed (extensive). Seropositivity for A. phagocytophilum in only one animal suggests the occurrence of a cross-serological reaction with another agent of the genus Anaplasma.

Resumo O gene msp4 de A. marginale é unicodon, estável e pouco heterogêneo, sendo considerado como um marcador útil para caracterização filogeográfica desta bactéria. Este trabalho teve como objetivo analisar a filogeografia de A. marginale com base no gene msp4 em bovinos de corte do Pantanal Brasileiro, comparativamente a outra regiões do mundo. Alíquotas de sangue foram colhidas de 400 bovinos (200 vacas e 200 bezerros) em cinco propriedades de cria e recria extensiva. Como resultado, 56,75% (227/400) mostraram-se positivas para A. marginale pela qPCR para o gene msp1β e destas, 8,37% (19/227) amostras foram positivas na PCR convencional para o gene msp4. Na análise de distância Network, 14 sequências do Pantanal brasileiro foram agrupadas em um único grupo com as da Thailândia, Índia, Espanha, Colômbia, Paraná (Brasil), México, Portugal, Argentina, China, Venezuela, Austrália, Italia e Minas Gerais (Brasil). Dentre 68 sequências do Brasil e do mundo, constatou-se a presença de 15 genótipos, sendo o genótipo número um (#1) o mais distribuído. As sequências msp4 de A. marginale detectadas em bovinos de corte no Pantanal brasileiro apresentaram baixo polimorfismo com formação de dois genogrupos filogeneticamente relacionados àqueles encontrados em ruminantes de países das América do Sul e Central, Europa e Ásia.

Abstract The msp4 gene of A. marginale is unicodon, stable and mostly homogeneous, being considered as a useful marker for phylogeographic characterization of this bacterium. The objective of this work was to analyze the phylogeography of A. marginale based on the msp4 gene in beef cattle from the Brazilian Pantanal, compared to those found in other regions worldwide. The blood samples investigated were collected from 400 animals (200 cows and 200 calves) reared in five extensive breeding farms in this region. The results indicated that of the evaluated samples, 56.75% (227/400) were positive for A. marginale based on the msp1β gene by quantitatitve PCR (qPCR), while 8.37% (19/227) were positive for the msp4 gene in the conventional PCR. In the Network distance analysis, 14 sequences from the Brazilian Pantanal were grouped into a single group with those from Thailand, India, Spain, Colombia, Parana (Brazil), Mexico, Portugal, Argentina, China, Venezuela, Australia, Italy and Minas Gerais (Brazil). Among 68 sequences from Brazil and the world, 15 genotypes were present while genotype number one (#1) was the most distributed worldwide. Both Splitstree and network analyses showed that the A. marginale msp4 sequences detected in beef cattle from the Brazilian Pantanal showed low polymorphism, with the formation of one genogroup phylogenetically related to those found in ruminants from South and Central America, Europe, and Asia.

Abstract Germline mutations in BRCA1 and BRCA2 (BRCA) genes confer high risk of developing cancer, especially breast and ovarian tumors. Since the cloning of these tumor suppressor genes over two decades ago, a significant amount of research has been done. Most recently, monoallelic loss-of-function mutations in PALB2 have also been shown to increase the risk of breast cancer. The identification of BRCA1, BRCA2 and PALB2 as proteins involved in DNA double-strand break repair by homologous recombination and of the impact of complete loss of BRCA1 or BRCA2 within tumors have allowed the development of novel therapeutic approaches for patients with germline or somatic mutations in said genes. Despite the advances, especially in the clinical use of PARP inhibitors, key gaps remain. Now, new roles for BRCA1 and BRCA2 are emerging and old concepts, such as the classical two-hit hypothesis for tumor suppression, have been questioned, at least for some BRCA functions. Here aspects regarding cancer predisposition, cellular functions, histological and genomic findings in BRCA and PALB2-related tumors will be presented, in addition to an up-to-date review of the evolution and challenges in the development and clinical use of PARP inhibitors.

Resumo O objetivo do trabalho foi avaliar a influência do sexo, da sazonalidade e de infecções por Trypanosoma cruzi e Trypanosoma evansi nos valores hematológicos de lobinhos (Cerdocyon thous) que habitam o Pantanal Sul-Mato-Grossense. Entre novembro de 2015 e outubro de 2016, foram amostrados 48 lobinhos. Os valores hematológicos mensurados foram as contagens de eritrócitos e leucócitos, volume globular e volume corpuscular médio. Consideramos como parasitados os animais positivos para T.cruzi e T. evansi em qualquer um dos testes diagnósticos utilizados. Observamos que sete (14.5%) lobinhos encontraram-se parasitados somente por T.cruzi, sete por T. evansi, e nove (19%) estavam coinfectados. Os animais parasitados por T.cruzi mostraram um aumento significativo das contagens de leucócitos (14.7 x 103) em relação aos animais não parasitados (10.4 x 103), parasitados por T. evansi (12.4 x 103) e coinfectados (12.9 x 103). Observamos diferença significativa em relação aos valores médios de eritrócitos e volume corpuscular médio entre os períodos de cheia e seca: 2.6 x 106 e 165, e 3.4 x 106 e 132, respectivamente. Todos apresentaram macrocitose não confirmada morfologicamente.

Abstract The aim of this work was to evaluate the influence of sex, seasonality and infections by Trypanosoma cruzi and Trypanosoma evansi on the hematological values of crab-eating foxes (Cerdocyon thous) that inhabit the Pantanal from Mato Grosso do Sul. Between November of 2015 and October of 2016, 48 animals were sampled. The hematological values measured were erythrocyte and leukocyte counts, globular volume and mean corpuscular volume. We considered as parasitized the positive animals for T.cruzi and T. evansi in any of diagnostic tests used. We observed that seven (14.5%) of the crab-eating foxes were parasitized only by T.cruzi, seven by T. evansi, and nine (19%) were coinfected. The parasitized animals by T.cruzi showed a significant increase in leukocyte counts (14.7 x 103) compared to non-parasitized animals (10.4 x 103), parasitized by T. evansi (12.4 x 103) and coinfected (12.9 x 103). We observed a significant difference in relation to mean erythrocyte and mean corpuscular volume values between wet and dry periods: 2.6 x 106 and 165, and 3.4 x 106 and 132, respectively. All presented morphologically unconfirmed macrocytosis.

Resumo Após uma revisão na literatura vimos que o Problema Restrito de Três Corpos (PRTC) é um tema com algumas publicações voltadas, principalmente, para a resolução do problema matemático, com muito poucas aplicações em situações reais e contextualizadas. Além disso, ao analisarmos alguns livros textos de mecânica clássica utilizados nos cursos de física, observamos que o PRTC não é desenvolvido e explorado didaticamente. Apoiados nessas análises o nosso objetivo neste trabalho é apresentar, desenvolver e aplicar o PRTC para simular as órbitas de dois astros do sistema solar: a do asteroide troiano da Terra, 2010 TK7, e a libração de Plutão devido a Netuno. Para resolver as equações diferencias do PRTC utilizamos o método de Cauchy em um código escrito na linguagem de programação Python. Nos dois casos estudados conseguimos verificar as trajetórias, valores e comportamentos orbitais dos astros. Por isso, entendemos que o PRTC pode ser aplicado para simular o movimento de diversos astros do sistema solar e, consequentemente, ser utilizado como um recurso pedagógico para contextualizar conceitos de mecânica celeste e dinâmica orbital. Uma vez que ele se relaciona com o estudo das equações diferencias, com a gravitação universal e com os métodos numéricos.

Abstract After a review in the literature we have seen that the Restricted Three-Body Problem (RTBP) is a topic with some publications focused, primarily, on resolution of the mathematical problem, with very few application in real and contextualized situations. In addition, in analyzing textbooks of classical mechanics we observed that the RTBP is not developed and didactically explored. Based on this facts, our objective in this work is to present, develop and apply the RTBP to simulate the orbits of two objects in the solar system : motion of Earth-trojan asteroid 2010 TK7 and libration of pluto due to Neptune. To solve differential equations of RTBP we uses the cauchy method in a code written in the Python programming language. With this work we hope to demonstrate that the study of the RTBP can be used to contextualize some concepts of celestial mechanics and orbital dynamics.Since it is related to the study of differential equations, with universal gravitation and numerical methods.

Abstract Germline mutations in the TP53 gene are associated with Li-Fraumeni and Li-Fraumeni-Like Syndromes, characterized by increased predisposition to early-onset cancers. In Brazil, the prevalence of the TP53-p.R337H germline mutation is exceedingly high in the general population and in cancer-affected patients, probably as result of a founder effect. Several genotyping methods are used for the molecular diagnosis of LFS/LFL, however Sanger sequencing is still considered the gold standard. We compared performance, cost and turnaround time of Sanger sequencing, PCR-RFLP, TaqMan-PCR and HRM in the p.R337H genotyping. The performance was determined by analysis of 95 genomic DNA samples and results were 100% concordant for all methods. Sequencing was the most expensive method followed by TaqMan-PCR, PCR-RFLP and HRM. The overall cost of HRM increased with the prevalence of positive samples, since confirmatory sequencing must be performed when a sample shows an abnormal melting profile, but remained lower than all other methods when the mutation prevalence was less than 2.5%. Sequencing had the highest throughput and the longest turnaround time, while TaqMan-PCR showed the lowest turnaround and hands-on times. All methodologies studied are suitable for the detection of p.R337H and the choice will depend on the application and clinical scenario.

A infecção simultânea por parasitas de diferentes espécies pode resultar em alterações imprevisíveis. O presente estudo avaliou a patologia de cães naturalmente coinfectados por Leishmania infantum e Ehrlichia canis. A saúde dos cães foi investigada pelas análises histopatológicas, hematológicas e bioquímicas de 21 cães infectados somente por L. infantum e 22 cães coinfectados por L. infantum e E. canis. Observou-se uma reação inflamatória crônica, predominantemente linfohistioplasmocítica, na pele dos dois grupos. A plasmocitose, encontrada nos tecidos linfóides, provavelmente estava relacionada com a hipergamaglobulinemia observada em todos os cães amostrados. A desorganização da matriz extracelular da derme da região inguinal e da orelha, demonstrada pela substituição das fibras de colágeno espessas por fibras finas, foi relacionada com o grau de reação inflamatória, independente da presença de parasitas. Ainda, observamos duas vezes mais animais do grupo coinfectado apresentando formas amastigotas na pele de orelha pela histopatologia comparado ao número de cães infectados apenas por Leishmania, tornando-os desta forma mais infectivos aos vetores. Nossos resultados ressaltam que a saúde de cães coinfectados estava severamente comprometida devido aos altos níveis de proteína plasmática total, globulinas, fosfatase alcalina, creatina quinase e anemia acentuada.

Different parasites that commonly occur concomitantly can influence one another, sometimes with unpredictable effects. We evaluated pathological aspects of dogs naturally co-infected with Leishmania infantum and Ehrlichia canis. The health status of the dogs was investigated based on histopathological, hematological and biochemical analyses of 21 animals infected solely with L. infantum and 22 dogs co- infected with L. infantum and E. canis. The skin of both groups showed chronic, predominantly lymphohistioplasmacytic inflammatory reaction. The plasmacytosis in the lymphoid tissues was likely related with the hypergammaglobulinemia detected in all the dogs. The disorganization of extracellular matrix found in the reticular dermis of the inguinal region and ear, characterized by the substitution of thick collagen fibers for thin fibers, was attributed to the degree of inflammatory reaction, irrespective of the presence of parasites. In addition, the histopathological analysis revealed that twice as many dogs in the co-infected group presented Leishmania amastigotes in the ear skin than those infected solely with Leishmania, increasing the possibility of becoming infected through sand fly vectors. Our findings highlight the fact that the health of dogs infected concomitantly with L. infantum and E. canis is severely compromised due to their high levels of total plasma protein, globulins, alkaline phosphatase and creatine kinase, and severe anemia.

Os autores descrevem sua experiência em recanalização tubária videolaparoscópica pós-laqueadura em 10 pacientes, tratadas no período de junho de 1994 a fevereiro de 1996. A duração da 1ª cirurgia foi de 4 horas e 30 minutos e da última foi de 2 horas e 30 minutos. A metade das recanalizações foi ístmo-ístmica. O tamanho mínimo das tubas restantes foi de 4 cm de cada lado e os fios utilizados para a sutura foram o 7-0 e 6-0 monofilamentares, derivados do ácido poliglicólico. A permeabilidade tubária foi verificada pela histerossalpingografia 3 meses após a cirurgia e, como resultado, 88, 8% das tubas operadas estavam permeáveis. Quatro das pacientes consideradas aptas para engravidar ficaram grávidas durante o período de seguimento. O tempo médio de permanência hospitalar foi de 24 horas. Não houve complicações cirúrgicas ou anestésicas.

The authors describe their experience with videolaparoscopic tubal anastomosis in 10 selected patients operated from June 1994 to February 1996. The time of the first surgery was 4 hours and 30 minutes and the last , 2 hours and 30 minutes. The time was different according to the change of auxiliary team. Half of the reanastomoses were isthmic- isthmic. The minimum size of the remaining tubes was 5cm on each side. We used 7-0 and 6-0 polyglycolic acid monofilament for suture. The tube patency was tested by hysterosalpingogram 3 months after surgery, and it was shown that 88.8% of the operated tubes were free. The patients considered able to become pregnant were followed up for a short period of time and 4 of them became pregnant. The hospitalization lasted 24 hours and there were no surgical or anesthetic complications.