Why not "do simple things in a simple way": Use of the Pap test as the first step in screening genetic stability for human cultured stem cell therapy?

The aim of this study was to analyze adipose tissue-derived mesenchymal stem cells (AT-MSCs) using the Pap test as a first screening step to evaluate genetic stability. Human adipose tissue from six healthy female donors was obtained from elective liposuction procedures. The cells were isolated, cultivated at P2/P3, characterized by flow cytometric analysis, and differentiation induced. The AT-MSCs were stained by Papanicolaou staining and analyzed according to the Bethesda classification, and viability-apoptosis relationships were evaluated. The results of the Pap test for Sample I indicated high-grade alterations consistent with genetic instability; for Samples II-V, atypical cells of undetermined significance; and for Sample VI, normal cells. These results demonstrate the potential of using the Pap test as an initial screening step to evaluate the genetic stability of cultured AT-MSCs and also suggest its use for other adherent cells such as embryonic stem cells or induced pluripotent stem cells

Evaluation of hybrid-short-coir-fibre-reinforced composites via full factorial design

A full factorial design (2231) has been used to investigate the effect of the useof sodium hydroxide fibre treatment, Portland cement and uniaxial pressure on the physical and mechanical properties of hybrid short coir fibre reinforced composites (HSCoirFRCs). The response variables considered in this work were the apparent density, porosity, tensile and flexural strength, the modulus of elasticity and the Charpy impact resistance. The alkali treatment contributed not only to reduce the apparent porosity, but also to increase the mechanical properties of the HSCoirFRCs. A reduction of the impact resistance and an increase of the apparent density was also identified after treatment. Cold pressing significantly affected the physical and mechanical properties of the HSCoirFRCs. Higher pressure levels enhanced the wettability of the fibres and, consequently, the mechanical performance of the composites. The incorporation of cement microparticles as a second reinforcement phase was however not effective, leading to decreased strength and an increased apparent density of the materials. The HSCoirFRC structure can be considered an economical and sustainable alternative for future secondary structural parts in lightweight transport applications

Incidence and Duration of Type-Specific Human Papillomavirus Infection in High-Risk HPV-Naïve Women: Results from the Control Arm of a Phase II HPV-16/18 Vaccine Trial

OBJECTIVES: Persistence of human papillomaviruses (HPVs) is necessary for cervical carcinogenesis. We evaluated incidence and duration of type-specific HPV infections and the influence of age and number of sexual partners. METHODS: Data were obtained from 553 women (15-25 years), who were seronegative and DNA-negative for high-risk HPV (HR-HPV) types and were enrolled in the placebo arm of a randomised trial of the HPV-16/18 vaccine (NCT00689741/NCT00120848). They were followed for 6.3 years. Cervicovaginal samples were self-collected at 3-month intervals for up to 27 months, and cervical samples were collected by clinicians at 6-month intervals until study end. Samples were tested for HPV types using a broad-spectrum PCR assay. Incidence rate ratios (RRs) and 95% CIs were used to estimate the association among age, sexual habits and HPV acquisition. RESULTS: Incidence rates (95% CI) using cervical samples were 11.8 (10.4 to 13.4) and 5.6 (4.7 to 6.6) per 1000 women-months for HR-HPVs and low-risk HPVs (LR-HPVs), respectively. Equivalent rates in combined cervicovaginal and cervical samples were 17.2 (15.4 to 19.2) and 6.9 (5.9 to 8.0), respectively. 54 per cent of HR-HPV types from combined cervicovaginal and cervical samples persisted for 1 year compared with 32.3% for LR-HPV types. The risk of acquiring any HPV infection was higher among women aged(RR=1.33, 95% CI 1.1 to 1.7) and women having \u3e1 sexual partner (RR=1.83, 95% CI 1.4 to 2.4) at baseline. CONCLUSIONS: HR-HPV infections were more common and lasted longer on average than LR-HPV infections. HPV acquisition was more common in younger women with multiple sexual partners

Recomendación de fertilizantes para Pinus y Eucalyptus en Corrientes, Argentina utilizando balance nutricional

■ Alcanzar metas de productividad deseadas en bosques implantados requiere suministrarleslas cantidades de nutrientes que estos demandan.■ La calibración de modelos de balance nutricional para especies forestales plantadas enArgentina es una alternativa promisora para recomendar fertilizantes con un fundamentomás científico y menos empírico.■ Estos modelos permiten entender cuando es preciso fertilizar y con qué dosis según cadasituación en busca de optimizar el uso de recursos y mantener la sustentabilidad de laproducción forestal.Fil: Stahringer, Nicolas I.. Universidade Federal de Viçosa.; BrasilFil: Neves, Julio C. L.. Universidade Federal de Viçosa.; BrasilFil: Paulucio, R. B.. Universidade Federal de Viçosa.; BrasilFil: Teixeira, José L.. Universidade Federal de Viçosa.; BrasilFil: Sussini, Paolo Adrián. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Chaco-Formosa. Estación Experimental Agropecuaria Colonia Benítez; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste; ArgentinaFil: Argüelles, R. T.. Universidad Nacional del Sur. Departamento de Agronomía; ArgentinaFil: Chamorro, Luis Leandro. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentin

Leucograma de bezerros Holstein sadios no primeiro mês de vida

To establish reference values and to assess the influence of age on the leukograms of healthy Holstein calves, blood samples were obtained from 300 animals. These samples were distributed equally (n=20) among 15 experimental groups according to age: birth to 8 hours, 9 to 16 hours, 17 to 24 hours, 2 days, 3 days, 4 days, 5 days, 6 to 7 days, 8 to 9 days, 10 to 11 days, 12 to 13 days, 14 to 15 days, 16 to 20 days, 21 to 25 days and 26 to 30 days of age. The maximum numbers of leukocytes (9,305.0/mL), segmented neutrophils (6,551.2/mL) and total neutrophils (6,678.3/mL) were noted within the first 8 hours of life, while band neutrophils peaked in number (133.3/mL) between 9 and 16 hours after birth. Meanwhile, the maximum total lymphocyte (4,992.1/µL) and typical lymphocyte (4,686.1/µL) counts occurred between 21 and 25 days, whereas atypical lymphocytes (388.5/µL) reached their maximum number between 26 and 30 days, demonstrating an inversion of the neutrophil:lymphocyte ratio over time. Thus, the influence of age on the leukocyte count of the evaluated calves was verified. The release of endogenous corticosteroids during labor or at birth may contribute to this variation in leukograms with age.Com a finalidade de estabelecer os valores de referência e de avaliar a influência do fator etário sobre o leucograma de bezerras sadias, da raça Holandesa, utilizaram-se amostras de sangue de 300 animais, distribuídos com igual número (n=20) por 15 grupos experimentais, de acordo com a idade: do nascimento até 8 horas de vida; de 9 até 16 horas; de 17 até 24 horas; 2 dias de idade; 3 dias; 4 dias; 5 dias; 6 a 7 dias; 8 a 9 dias; 10 a 11 dias; 12 a 13 dias; 14 a 15 dias; de 16 a 20 dias; de 2l a 25 dias; e de 26 a 30 dias de vida. Puderam-se observar valores máximos nas primeiras oito horas de vida para leucócitos (9305,0/µL), neutrófilos segmentados (6551,2/µL) e neutrófilos totais (6678,3/µL); entre 9 a 16 horas pós-nascimento para os neutrófilos bastonetes (133,3/µL). Para os linfócitos totais (4992,1/µL) e os linfócitos típicos (4686,1/µL) os valores máximos ocorreram entre 21 e 25 dias, e para os linfócitos atípicos (388,5/µL), entre 26 e 30 dias, demonstrando-se uma inversão na relação neutrófilo: linfócito, com o decorrer dos dias de vida. Constatou-se a da influência do fator etário no leucograma dos bezerros avaliados, no entanto acredita-se que a liberação de corticoides endógenos no momento do parto/nascimento, contribuíram para esta variação

Genetic diversity and phylogenetic relationships of coevolving symbiont-harboring insect trypanosomatids, and their neotropical dispersal by invader African blowflies (Calliphoridae)

This study is about the inter- and intra-specific genetic diversity of trypanosomatids of the genus Angomonas, and their association with Calliphoridae (blowflies) in Neotropical and Afrotropical regions. Microscopic examination of 3,900 flies of various families, mostly Calliphoridae, revealed that 31% of them harbored trypanosomatids. Small subunit rRNA (SSU rRNA) barcoding showed that Angomonas predominated (46%) over the other common trypanosomatids of blowflies of genera Herpetomonas and Wallacemonas. Among Angomonas spp., A. deanei was much more common than the two-other species, A. desouzai and A. ambiguus. Phylogenetic analyses based on SSU rRNA, glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) and internal transcribed spacer rDNA (ITS rDNA) sequences revealed a marked genetic diversity within A. deanei, which comprised four infraspecific genotypes (Dea1– Dea4), and four corresponding symbiont genotypes (Kcr1–Kcr4). Host and symbiont phylogenies were highly congruent corroborating their co-divergence, consistent with host-symbiont interdependent metabolism and symbiont reduced genomes shaped by a long coevolutionary history. We compared the diversity of Angomonas/symbionts from three genera of blowflies, Lucilia, Chrysomya and Cochliomyia. A. deanei, A. desouzai, and A. ambiguus were found in the three genera of blowflies in South America. In Africa, A. deanei and A. ambiguus were identified in Chrysomya. The absence of A. desouzai in Africa and its presence in Neotropical Cochliomyia and Lucilia suggests parasite spillback of A. desouzai into Chrysomya, which was most likely introduced four decades ago from Africa into the Neotropic. The absence of correlation between parasite diversity and geographic and genetic distances, with identical genotypes of A. deanei found in the Neotropic and Afrotropic, is consistent with disjunct distribution due to the recent human-mediated transoceanic dispersal of Angomonas by Chrysomya. This study provides the most comprehensive data gathered so far on the genetic repertoires of a genus of trypanosomatids found in flies from a wide geographical range.The PROAFRICA, INCT-EPIAMO, and PROSUL programs of CNPq, PNIPB of Capes, and FAPESP (Process 2016/07487-0). CAPEs (PNPD) granted a postdoctoral scholarship to TB.http://www.frontiersin.org/Microbiologyam2018Veterinary Tropical Disease

New insights into the evolution of the Trypanosoma cruzi clade provided by a new trypanosome species tightly linked to Neotropical Pteronotus bats and related to an Australian lineage of trypanosomes

Abstract\ud \ud Background\ud Bat trypanosomes are implicated in the evolution of the T. cruzi clade, which harbours most African, European and American trypanosomes from bats and other trypanosomes from African, Australian and American terrestrial mammals, including T. cruzi and T. rangeli, the agents of the American human trypanosomiasis. The diversity of bat trypanosomes globally is still poorly understood, and the common ancestor, geographical origin, and evolution of species within the T. cruzi clade remain largely unresolved.\ud \ud \ud Methods\ud Trypanosome sequences were obtained from cultured parasites and from museum archived liver/blood samples of bats captured from Guatemala (Central America) to the Brazilian Atlantic Coast. Phylogenies were inferred using Small Subunit (SSU) rRNA, glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH), and Spliced Leader (SL) RNA genes.\ud \ud \ud Results\ud Here, we described Trypanosoma wauwau n. sp. from Pteronotus bats (Mormoopidae) placed in the T. cruzi clade, then supporting the bat-seeding hypothesis whereby the common ancestor of this clade likely was a bat trypanosome. T. wauwau was sister to the clade T. spp-Neobats from phyllostomid bats forming an assemblage of trypanosome species exclusively of Noctilionoidea Neotropical bats, which was sister to an Australian clade of trypanosomes from indigenous marsupials and rodents, which possibly evolved from a bat trypanosome. T. wauwau was found in 26.5 % of the Pteronotus bats examined, and phylogeographical analysis evidenced the wide geographical range of this species. To date, this species was not detected in other bats, including those that were sympatric or shared shelters with Pteronotus. T. wauwau did not develop within mammalian cells, and was not infective to Balb/c mice or to triatomine vectors of T. cruzi and T. rangeli.\ud \ud \ud Conclusions\ud \ud Trypanosoma wauwau n. sp. was linked to Pteronotus bats. The positioning of the clade T. wauwau/T.spp-Neobats as the most basal Neotropical bat trypanosomes and closely related to an Australian lineage of trypanosomes provides additional evidence that the T. cruzi clade trypanosomes likely evolved from bats, and were dispersed in bats within and between continents from ancient to unexpectedly recent times.This work was supported by grants from the Brazilian agencies CNPq (PROSUL,\ud PRAFRICA and PROTAX), CAPES (PNIPB and PNPD) and FAPESP. The analysis of\ud bats from Central America, Suriname and Guyana was supported by grant\ud ‘Investissements d’Avenir’ from the Agence Nationale de la Recherche, Canada\ud (ANR-10-LABX-25-01). Archived samples from Brazilian Pteronotus were donated\ud to ACP by VC Tavares, A Césari, PA Rocha, FM Martins, MOG Lopes, CS Bernabé,\ud TG Oliveira, E Gonçalves and M Marcos. We are grateful to many student from\ud USP and researches of other universities for the inestimable help in the\ud fieldworks. We also thanks JA Rosa for the generous contribution with\ud triatomines from the insectary of UNESP-Araraquara, and CE Jared and MM\ud Antoniazzi for the access to electron microscopic facilities of the Institute\ud Butantan, Brazil. Luciana Lima is postdoctoral fellow sponsored by FAPESP, and\ud Oneida Espinosa-Álvarez is recipient of a PhD fellowship from CNPq (PROTAX)

Natural history of progression of HPV infection to cervical lesion or clearance: analysis of the control arm of the large, randomised PATRICIA study

Background: The control arm of PATRICIA (PApillomaTRIal against Cancer In young Adults, NCT00122681) was used to investigate the risk of progression from cervical HPV infection to cervical intraepithelial neoplasia (CIN) or clearance of infection, and associated determinants. Methods and Findings: Women aged 15-25 years were enrolled. A 6-month persistent HPV infection (6MPI) was defined as detection of the same HPV type at two consecutive evaluations over 6 months and clearance as ≥2 type-specific HPV negative samples taken at two consecutive intervals of approximately 6 months following a positive sample. The primary endpoint was CIN grade 2 or greater (CIN2+) associated with the same HPV type as a 6MPI. Secondary endpoints were CIN1+/CIN3+ associated with the same HPV type as a 6MPI; CIN1+/CIN2+/CIN3+ associated with an infection of any duration; and clearance of infection. The analyses included 4825 women with 16,785 infections (3363 womenwith 6902 6MPIs). Risk of developing a CIN1+/CIN2+/CIN3+ associated with same HPV type as a 6MPI varied with HPV type and was significantly higher for oncogenic versus non-oncogenic types. Hazard ratios for development of CIN2+ were 10.44 (95% CI: 6.96-15.65), 9.65 (5.97-15.60), 5.68 (3.50-9.21), 5.38 (2.87-10.06) and 3.87 (2.38-6.30) for HPV-16, HPV-33, HPV-31, HPV-45 and HPV-18, respectively. HPV-16 or HPV-33 6MPIs had ~25-fold higher risk for progression to CIN3+. Previous or concomitant HPV infection or CIN1+ associated with a different HPV type increased risk. Of the different oncogenic HPV types, HPV-16 and HPV-31 infections were least likely to clear. Conclusions: Cervical infections with oncogenic HPV types increased the risk of CIN2+ and CIN3+. Previous or concomitant infection or CIN1+ also increased the risk. HPV-16 and HPV-33 have by far the highest risk of progression to CIN3+, and HPV-16 and HPV-31 have the lowest chance of clearance

Prevençao Secundária da Morte Súbita: Importância do Marcapasso Definitivo Prévio ao Implante de Cardioversor-desfibrilador Implantável (CDI) na Sobrevida de Pacientes com Miocardiopatia Chagásica

Objetivo: Avaliar a importância clínica da presença de marcapasso definitivo (MPD) previamente ao implante de CDI, considerando variáveis clínicas e epidemiológicas. Métodos: Dos 321 pacientes do banco de dados de CDI de nossa instituiçao, foram selecionados 275 submetidos a implante de CDI para prevençao secundária de morte súbita cardíaca (MSC), agrupados de acordo com a cardiomiopatia de base e a presença de MPD prévio ao implante de CDI. As variáveis analisadas foram: sexo, idade, CF-NYHA, medicaçoes, ritmo cardíaco, FEVE e TVNS. Para análise estatística, utilizou-se o método de Kaplan-Meier e o teste de log-rank. Resultados: A amostra reduzida de pacientes com cardiomiopatia nao chagásica e MPD prévio ao implante de CDI (N=6) nao permitiu análises estatísticas consistentes. Nos pacientes com cardiomiopatia chagásica (CCH), as características de base nos subgrupos com e sem MPD prévio foram estatisticamente semelhantes, exceto pela maior prevalência de TVNS no subgrupo sem MPD prévio. Nos pacientes com CCH, a comparaçao das curvas de sobrevida dos subgrupos com e sem MPD prévio evidenciou uma diferença significativa (p&0,05). A probabilidade de sobrevida no final do primeiro e terceiro anos foi de 78% e 39% nos pacientes com MPD prévio (N=18) e 87% e 58% nos pacientes sem MPD prévio (N=72). Conclusoes: Nos pacientes com CCH submetidos a implante de CDI para prevençao secundária de MSC, a presença de MPD previamente ao implante de CDI apresentou prevalência elevada (20%) e associou-se a um pior prognóstico