Effects on the Properties of Self-Compacting Cement Paste (PAA) with the Addition of Superabsorbent Polymer.

The addition of Superabsorbent Polymer (SAP) decreases the effect of autogenous shrinkage present in pastes, mortars, and concretes. In this study we investigated the influence of the addition of SAP in self-compacting cement paste mixtures. Eighteen 5 × 10 cylindrical specimens were molded in all, three for each mixture (CPII base, CPII 0.15%SAP/600µm, CPII 0.15%SAP/800 µm, CPV base, CPV 0.15%SAP/600 µm, CPV 0.15%SAP/800 µm). Two types of cement were tested, CP II-Z and CP V-ARI with 0.15% of weight replaced per two diameters of SAP (600 µm and 800 µm). The samples followed the standards required. Mini slump tests were carried out in the fresh state, and uniaxial compressive strength, elastic modulus, specific mass, absorption, and air content in the hardened state after 28 days. The results obtained show the SAP is high indicated to replaced cement in small % of weight i/to fresh and hardened paste. Likewise, the group mix n° 3 composed of CPII 0.15% of SAP with 800 µm diameter presented the best result.

Physical and Chemical Properties of Waste Foundry Exhaust Sand for Use in Self-Compacting Concrete.

The reuse of waste in civil construction brings environmental and economic benefits. However, for these to be used in concrete, it is necessary a previous evaluation of their physical and chemical characteristics. Thus, this study aimed to characterize and analyze the waste foundry exhaust sand (WFES) for use in self-compacting concrete (SCC). Foundry exhaust sand originates from the manufacturing process of sand molds and during demolding of metal parts. It is a fine sand rich in silica in the form of quartz collected by baghouse filter. Characterization of WFES was conducted through laser granulometry, scanning electron microscopy (SEM) in the energy dispersive spectroscopy (EDS) mode, X-ray diffraction (XRD), X-ray fluorescence (XRF), Fourier transform infrared spectroscopy (FTIR), thermogravimetry (TG) and derivative thermogravimetry (DTG) techniques. The waste was classified as non-hazardous and non-inert, with physical and chemical properties suitable for use in SCC composition, as fine aggregate or mineral addition. Five mixtures of SCC were developed, in order to determine the waste influence in both fresh and hardened concrete. The properties in the fresh state were reached. There was an increase in compressive strength and sulfate resistance, a decrease in water absorption of self-compacting concrete by incorporating WFES as 30% replacement.

Bibliometric analysis of studies involving e-waste: a critical review.

The correct destination of waste is an essential factor for sustainable development. Electronic waste, which is very toxic, is the type of waste with the highest rate of increase in its generation. For these reasons, the amount of research on this topic increases year by year, as shown by the literature review carried out by this study. This review aims to identify the main characteristics and proposals of the main study on electronic waste and verify how Brazil is inserted globally in the research on e-waste. Another objective is to suggest a path for researchers who want to start research on e-waste by identifying the keywords most used in the analyzed articles. The results evidenced that the most published countries on the subject are China, the USA, and India. Brazil is in a position of little prominence concerning its research production on the subject. Most of the central studies we analyzed use case study and literature review as the research method. Among the 44 articles analyzed, only one proposed a destination for e-waste. It highlights the need for more research focusing on the environmentally correct destination of e-waste. A good way to start a search on electronic waste is to use the keywords identified in this study, especially those used most frequently in the analyzed articles.

Individual selection of the first backcross generation of passion fruit potentially resistant to the fruit woodiness disease.

The REML/BLUP procedure has been successfully used for genetic progress through individual selection of high-yielding passion fruit genotypes resistant to the Cowpea aphid-borne mosaic virus. This study was thus developed to estimate genetic parameters and predict the gain obtained from individual selection of genotypes in a population derived from backcrosses in passion fruit. The experiment was set up as a randomized block design with four replicates, involving five full-sib families (genotypes from the first backcross). Variance components and the genetic values were estimated for eight agronomic traits via the REML/BLUP procedure. For all traits, genotypic variance between the genotypes from the first backcross showed little contribution to the phenotypic variance. The low heritability estimates obtained for the traits are overcome via individual BLUP estimates. Therefore, it was possible to obtain considerable gains with individual selection for the variables fruit length, average fruit weight, and pulp weight (19.50 to 14.04%; 22.93 to 17.97%; and 10.08 to 7.95%, respectively). For the traits showing lower gains, it is possible to obtain gains indirectly by selecting genotypes for correlated traits. Because this population derives from the first backcross generation, agronomic traits still must be recovered.

NEK10 interactome and depletion reveal new roles in mitochondria.

BACKGROUND: Members of the family of NEK protein kinases (NIMA-related kinases) were described to have crucial roles in regulating different aspects of the cell cycle. NEK10 was reported to take part in the maintenance of the G2/M checkpoint after exposure to ultraviolet light. NEK1, NEK5, NEK2 and NEK4 proteins on the other hand have been linked to mitochondrial functions. METHODS: HEK293T cells were transfected with FLAG empty vector or FLAG-NEK10 and treated or not with Zeocin. For proteomic analysis, proteins co-precipitated with the FLAG constructs were digested by trypsin, and then analyzed via LC-MS/MS. Proteomic data retrieved were next submitted to Integrated Interactome System analysis and differentially expressed proteins were attributed to Gene Ontology biological processes and assembled in protein networks by Cytoscape. For functional, cellular and molecular analyses two stable Nek10 silenced HeLa cell clones were established. RESULTS: Here, we discovered the following possible new NEK10 protein interactors, related to mitochondrial functions: SIRT3, ATAD3A, ATAD3B, and OAT. After zeocin treatment, the spectrum of mitochondrial interactors increased by the proteins: FKBP4, TXN, PFDN2, ATAD3B, MRPL12, ATP5J, DUT, YWHAE, CS, SIRT3, HSPA9, PDHB, GLUD1, DDX3X, and APEX1. We confirmed the interaction of NEK10 and GLUD1 by proximity ligation assay and confocal microscopy. Furthermore, we demonstrated that NEK10-depleted cells showed more fragmented mitochondria compared to the control cells. The knock down of NEK10 resulted further in changes in mitochondrial reactive oxygen species (ROS) levels, decreased citrate synthase activity, and culminated in inhibition of mitochondrial respiration, affecting particularly ATP-linked oxygen consumption rate and spare capacity. NEK10 depletion also decreased the ratio of mtDNA amplification, possibly due to DNA damage. However, the total mtDNA content increased, suggesting that NEK10 may be involved in the control of mtDNA content. CONCLUSIONS: Taken together these data place NEK10 as a novel regulatory player in mitochondrial homeostasis and energy metabolism.

XPD/ERCC2 mutations interfere in cellular responses to oxidative stress.

Nucleotide excision repair (NER) is a conserved, flexible mechanism responsible for the removal of bulky, helix-distorting DNA lesions, like ultraviolet damage or cisplatin adducts, but its role in the repair of lesions generated by oxidative stress is still not clear. The helicase XPD/ERCC2, one of the two helicases of the transcription complex IIH, together with XPB, participates both in NER and in RNA pol II-driven transcription. In this work, we investigated the responses of distinct XPD-mutated cell lines to the oxidative stress generated by photoactivated methylene blue (MB) and KBrO3 treatments. The studied cells are derived from patients with XPD mutations but expressing different clinical phenotypes, including xeroderma pigmentosum (XP), XP and Cockayne syndrome (XP-D/CS) and trichothiodystrophy (TTD). We show by different approaches that all XPD-mutated cell lines tested were sensitive to oxidative stress, with those from TTD patients being the most sensitive. Host cell reactivation (HCR) assays showed that XP-D/CS and TTD cells have severely impaired repair capacity of oxidised lesions in plasmid DNA, and alkaline comet assays demonstrated the induction of significantly higher amounts of DNA strand breaks after treatment with photoactivated MB in these cells compared to wild-type cells. All XPD-mutated cells presented strong S/G2 arrest and persistent γ-H2AX staining after photoactivated MB treatment. Taken together, these results indicate that XPD participates in the repair of lesions induced by the redox process, and that XPD mutations lead to differences in the response to oxidatively induced damage.

Association between overweight and consumption of ultra-processed food and sugar-sweetened beverages among vegetarians / Associação entre o excesso de peso e o consumo de alimentos ultraprocessados e bebidas açucaradas por diferentes grupos vegetarianos

ABSTRACT Objective: To assess the consumption of ultra-processed food and sugar-sweetened beverages and to identify the association of this consumption with overweight among vegetarians. Methods: A cross-sectional study with a convenience sampling method was conducted. Data were collected using an online questionnaire from 8/24/2015 to 10/8/2015. Subjects were male and female vegetarians aged >16 years. Using a food frequency questionnaire, we assessed the weekly consumption of ultra-processed food and sugar-sweetened beverages and described the frequency of daily consumption overall and according to type of vegetarianism. The association between overweight and excessive daily intake of ultra-processed food was analyzed by multiple logistic regression (OR [95CI%]). Results: Information was retrieved from 503 individuals (29.8±8.5 years old); 83.7% were women. The most frequent types of vegetarianism in our sample were ovo-lacto (45.5%) and vegan (41.7%), and the median time of vegetarianism was 5.3 years. The consumption of sugar-sweetened beverages (≥2x/day) and ultra-processed food (≥3x/day) was 21.0% and 16.0%, respectively, and regarding the different vegetarianism types, vegans showed the lowest frequency of excessive daily sugar-sweetened beverages and ultra-processed food consumption. In the multivariable analysis, consumption of ultra-processed food ≥3x/day (2.33 [1.36-4.03]), male sex (1.73 [1.01-2.96]), age ≥35 years (2.03 [1.23-3.36]) and not preparing one's food (1.67 [0.95-2.94]) were independently associated with overweight. Conclusion: Although vegetarianism is frequently associated with a healthier diet and, consequently, prevention of poor health outcomes, this study found that the excessive consumption of sugar-sweetened beverages and ultra-processed food was associated with overweight.

RESUMO Objetivo: Avaliar o consumo de alimentos ultraprocessados e de bebidas açucaradas entre indivíduos vegetarianos e identificar sua associação com o excesso de peso. Métodos: Estudo transversal com método amostral por conveniência, sendo os dados coletados por meio de questionário virtual entre 24/8/2015 e 8/10/2015. A amostra foi composta por indivíduos vegetarianos de ambos os sexos com idade >16 anos. Estudou-se a frequência de consumo de alimentos ultraprocessados e bebidas açucaradas, avaliados por meio de questionário de frequência alimentar, sendo categorizados em função do consumo semanal e diário e do tipo de prática vegetariana. A associação entre excesso de peso e consumo excessivo de alimentos ultraprocessados foi realizada por meio de regressão logística múltipla (RC [IC95%]). Resultados: Participaram desta pesquisa 503 indivíduos (29,8±8,5 anos), com predominância do sexo feminino (83,7%). As principais práticas vegetarianas observadas foram a ovolactovegetariana (45,5%) e a vegana (41,7%), sendo a mediana de tempo de adesão ao vegetarianismo de 5,3 anos. Observou-se que o consumo de bebidas açucaradas (≥2x/dia) e alimentos ultraprocessados (≥3x/dia) foi, respectivamente, 21,0% e 16,0%; em relação aos outros grupos, os veganos apresentaram consumo inferior de alimentos ultraprocessados e bebidas açucaradas. No modelo múltiplo, identificou-se que consumo ≥3x/dia de alimentos ultraprocessados (2,33 [1,36-4,03]), sexo masculino (1,73 [1,01-2,96]), idade ≥35 anos (2,03 [1,23-3,36]) e não preparar as refeições em casa (1,67 [0,95-2,94]) se associaram independentemente com excesso de peso. Conclusão: Embora o vegetarianismo seja frequentemente associado à alimentação saudável e, consequentemente, à pre-venção de desfechos desfavoráveis a saúde, este estudo identificou frequência elevada do consumo diário excessivo de bebidas açucaradas e alimentos ultraprocessados e que este padrão de consumo se associou ao excesso de peso.

Cardiolipin is a key determinant for mtDNA stability and segregation during mitochondrial stress.

Mitochondria play a key role in adaptation during stressing situations. Cardiolipin, the main anionic phospholipid in mitochondrial membranes, is expected to be a determinant in this adaptive mechanism since it modulates the activity of most membrane proteins. Here, we used Saccharomyces cerevisiae subjected to conditions that affect mitochondrial metabolism as a model to determine the possible role of cardiolipin in stress adaptation. Interestingly, we found that thermal stress promotes a 30% increase in the cardiolipin content and modifies the physical state of mitochondrial membranes. These changes have effects on mtDNA stability, adapting cells to thermal stress. Conversely, this effect is cardiolipin-dependent since a cardiolipin synthase-null mutant strain is unable to adapt to thermal stress as observed by a 60% increase of cells lacking mtDNA (ρ0). Interestingly, we found that the loss of cardiolipin specifically affects the segregation of mtDNA to daughter cells, leading to a respiratory deficient phenotype after replication. We also provide evidence that mtDNA physically interacts with cardiolipin both in S. cerevisiae and in mammalian mitochondria. Overall, our results demonstrate that the mitochondrial lipid cardiolipin is a key determinant in the maintenance of mtDNA stability and segregation.

Estudo dos mecanismos moleculares do reparo de quebra de duplas fitas no DNA mitocondrial / Study of the molecular mechanisms of double-strand break repair in mitochondrial DNA

O DNA está constantemente exposto a danos causados tanto por agentes endógenos quanto exógenos. Estes podem causar diferentes tipos de lesões incluindo modificações de bases e do açúcar, além de quebras de fitas simples ou duplas. As quebras de duplas fitas, quando comparadas às demais, constituem as mais citotóxicas e podem resultar em deleções no DNA e instabilidade genética. Deleções no DNA mitocondrial (mtDNA) causam diversas doenças e estão envolvidas no processo de envelhecimento. No núcleo, as quebras de duplas fitas no DNA podem ser reparadas por recombinação homóloga (HR), ligação de pontas não homólogas (NHEJ) e anelamento de fita simples (SSA). No entanto, em mitocôndrias de células de mamíferos, o reparo de quebras de duplas fitas ainda não foi completamente caracterizado. Experimentos in vitro usando extratos mitocondriais de células de roedores mostraram que estes são capazes de reparar essas quebras, no entanto pouco é sabido sobre quais proteínas são responsáveis por cada etapa de reparo, bem como sua implicação na manutenção da integridade do genoma mitocondrial. Sendo assim, nesse trabalho investigamos a localização e função mitocondrial das proteínas ATM, Rad51, Rad52, Ku70/86 e DNA-PKCs, que são sabidamente envolvidas em reparo de quebras de duplas fitas no núcleo. Para identificar essas proteínas em mitocôndrias de células de mamíferos, mitocôndrias foram isoladas a partir de células da linhagem HEK293T, usando centrifugação diferencial seguida por gradiente de Percoll. Para as proteínas de recombinação homóloga, ATM e Rad51, imunodetectamos isoformas semelhantes em todos os compartimentos celulares. Já para a proteína Rad52 o mesmo anticorpo imunodetectou duas bandas distintas na mitocôndria ao passo que no núcleo foram quatro. Além disso, verificamos que baixos níveis de proteína Rad52, induzidos pela expressão de shRNA (short hairping RNA) específico, resultam em diminuição do número de cópias de mtDNA bem como acúmulo de deleções no genoma mitocondrial. Para as proteínas de NHEJ, DNA-PKCs e a subunidade Ku70, identificamos isoformas semelhantes em todos os compartimentos celulares. Já para a subunidade 86 do heterodímero Ku70/86 o anticorpo detectou, somente em mitocôndrias, uma banda menor de 50 kDa, a qual difere na região N-terminal da subunidade detectada no núcleo (86 KDa). Experimentos de co-imunprecitação de proteínas mostraram que essa isoforma menor compõe o heterodímero mitocondrial juntamente com a subunidade 70 (mtKu70/50) e que esse interage com DNA ligase III mitocondrial. Nossos resultados também mostraram que a estabilidade proteica de mtKu70/50 é regulada por ATM. Tratamento das células com peróxido de hidrogênio, que induz quebras de duplas fitas, aumentou a associação do heterodímero mtKu70/50 com o mtDNA, de forma independente de aumento da concentração proteica intra-mitocondrial. Já a diminuição dos níveis proteicos de Ku, induzida através de shRNA, resultou em diminuição do número de cópias de mtDNA e acumulo de danos nesse genoma. Extratos mitocondriais de células knockdown para Ku apresentaram menor atividade de reparo NHEJ em um ensaio in vitro, sugerindo que o acúmulo de danos nestas células é provavelmente devido a deficiências na via de NHEJ. Em conjunto, nossos dados sugerem que tanto HR quanto NHEJ operam em mitocôndrias. Além disso, a via de NHEJ mitocondrial utiliza o heterodímero mitocondrial Ku70/50 o qual está envolvido na manutenção do mtDNA. Ademais, nossos resultados mostram uma grande conservação molecular e funcional entre as vias de reparo de NHEJ e HR no núcleo e na mitocôndria, o que reforça sua importância para a manutenção da estabilidade genômica mitocondrial e, provavelmente a função mitocondrial

DNA is constantly exposed to damaging agents from both endogenous and exogenous sources. These can cause different types of DNA lesions that include base and sugar modifications and single and double strand breaks. DNA doublestrand breaks (DSBs) are among the most cytotoxic DNA lesions, which can result in deletions and genetic instability. Deletions in the mitochondrial DNA (mtDNA) cause numerous human diseases and drive normal aging. DSBs in the nuclear DNA are repaired by non-homologous DNA end joining (NHEJ), homologous recombination (HR) or Single Strand Annealing (SSA). Yet, repair of DSBs in mammalian mitochondria has not been fully characterized. Mitochondrial extracts from rodent cells are proficient in ligating DNA ends in vitro, but little is known about which proteins are responsible for each enzymatic step and its implication in mitochondrial genome maintenance. Thus, we investigated mitochondrial localization and function of DSBR (double strand break repair) proteins ATM, Rad51, Rad52, the Ku70/86 heterodimer and DNA-PKCs.To identify DSBR proteins in mammalian mitochondria, highly purified mitochondria from HEK293T cells were isolated using differential centrifugation followed by Percoll gradient. For HR proteins, we detected similar isoforms for ATM and Rad51 proteins in all cellular compartments. Two mitochondriaspecific isoforms of Rad52 were detected, while the same antibody detected four isoforms in the nucleus. In addition, lower Rad52 protein levels, induced by specific shRNA expression, result in decreased mtDNA copy number and accumulation of deleted mitochondrial genomes. For NHEJ proteins, similar isoforms of DNA-PKcs and the Ku70 subunit were detected in all cellular compartments. On the other hand, antibodies against the Ku86 subunit detected a smaller band in mitochondrial extracts (50 KDa), lacking the N-terminal region of the canonical isoform detected in the nucleus (86 KDa). The mitochondrial Ku70/50 heterodimer interacts with mitochondrial DNA ligase III, suggesting a role in DSBR. Moreover, stability of the mtKu heterodimer is regulated by ATM. Hydrogen peroxide treatment, which induces DSBs, increases mtKu70/50 association with the mtDNA and cells with reduced Ku levels, also induced by shRNA transfection, have lower mtDNA copy number and accumulate mtDNA damage. Moreover, mitochondrial extracts from Ku knockdown cells show lower NHEJ repair activity in an in vitro assay, suggesting that damage accumulation in these cells is likely due to deficiencies in NHEJ. Together, our data suggest that both HR and NHEJ operate in mitochondria. Also, mtNHEJ requires the Ku heterodimer and is involved in mtDNA maintenance. Moreover, our results indicate that there is a significant molecular and functional conservation between NHEJ and HR repair pathways in the nucleus and in mitochondria, which reinforces their importance for maintenance of mitochondrial genomic stability and, likely mitochondrial function

DivIVA-mediated polar localization of ComN, a posttranscriptional regulator of Bacillus subtilis.

ComN (YrzD) is a small, 98-amino-acid protein recently shown to be involved in the posttranscriptional control of the late competence comE operon in Bacillus subtilis. We show here that ComN localizes to the division site and cell poles in a DivIVA-dependent fashion. Yeast two-hybrid and glutathione S-transferase pulldown experiments showed that ComN interacts directly with DivIVA. ComN is not essential for the polar assembly of the core competence DNA uptake machinery. Nevertheless, polar localization of ComN should play some role in competence acquisition because delocalization of ComN leads to a small reduction in competence efficiency. We found that ComN promotes the accumulation of its target comE mRNA to septal and polar sites. Thus, we speculate that localized translation of ComE proteins may be required for efficient competence development. Our results underscore the versatility of DivIVA as a promoter of the differentiation of bacterial poles and demonstrate that the repertoire of polarly localized molecules in B. subtilis is broad, including a regulator of gene expression and its target mRNA. Moreover, our findings suggest that mRNA localization may play a role in the subcellular organization of bacteria.

Alto riesgo obstetrico (ARO), Trabajo de Metodologia de la Investigación

En la bibliografia incluye sitios WEB y fichas bibliográficas

Experiência de cárie e situaçäo de higiene oral em crianças dos orfanatos de Anápolis-GO / Caries experience and oral hygiene status among institutionalized children in Anápolis-GO, Brazil

O objetivo do presente estudo foi avaliar a condiçäo de saúde bucal das crianças mantidas nos orfanatos de Anápolis-GO. A populaçäo de estudo foi constituída de 168 crianças de ambos os sexos, na idade de 6 a 12 anos. A prevalência de cárie foi bastante elevada em todas as idades e apenas 5,3 por cento das crianças apresentaram-se livres de cárie. Aos 12 anos, o CPO-D foi 6,37. As necessidades de tratamento na dentiçäo decídua e permanente foram superiores às necessidades atendidas. A situaçäo de higiene oral (IHOS) mostrou-se deficiente para todas as idades. Conclui-se que o estado de saúde bucal deste grupo de crianças é precário e que há necessidade urgente de medidas de promoçäo de saúde bucal nas instituiçöes pesquisadas