The pinewood nematode, Bursaphelenchus xylophilus, in Madeira Island

The environmental conditions in Madeira Island are favorable for the presence and dissemination of the pinewood nematode (PWN), Bursaphelenchus xylophilus. Five hundred Pinus pinaster wood samples were collected in several forest areas and PWN was detected in 22.8 % of the samples. Bursaphelenchus xylophilus isolates from Madeira Island displayed the species-specific diagnostic characters. A morphological variation in the female tail terminus was detected. In most females, the tail presented a broadly rounded terminus and, occasionally, a digitate terminus with a terminal nipple-like extension resembling a mucro. PCR ITS-RFLP analysis revealed that Madeira Island isolates exhibited patterns specific to the species B. xylophilus and similar to virulent isolates. Amplified ITS regions were further sequenced and no genetic diversity was found for this genomic region among 17 Portuguese isolates (Madeira Island and Continental Portugal). Phylogenetic analysis revealed that Portuguese isolates grouped with isolates from China, Korea and one isolate from Japa

S100B chaperone multimers suppress the formation of oligomers during Aβ42 aggregation

Extracellular aggregation of the amyloid-β 1–42 (Aβ42) peptide is a major hallmark of Alzheimer’s disease (AD), with recent data suggesting that Aβ intermediate oligomers (AβO) are more cytotoxic than mature amyloid fibrils. Understanding how chaperones harness such amyloid oligomers is critical toward establishing the mechanisms underlying regulation of proteostasis in the diseased brain. This includes S100B, an extracellular signaling Ca2+-binding protein which is increased in AD as a response to neuronal damage and whose holdase-type chaperone activity was recently unveiled. Driven by this evidence, we here investigate how different S100B chaperone multimers influence the formation of oligomers during Aβ42 fibrillation. Resorting to kinetic analysis coupled with simulation of AβO influx distributions, we establish that supra-stoichiometric ratios of dimeric S100B-Ca2+ drastically decrease Aβ42 oligomerization rate by 95% and AβO levels by 70% due to preferential inhibition of surface-catalyzed secondary nucleation, with a concomitant redirection of aggregation toward elongation. We also determined that sub-molar ratios of tetrameric apo-S100B decrease Aβ42 oligomerization influx down to 10%, while precluding both secondary nucleation and, more discreetly, fibril elongation. Coincidently, the mechanistic predictions comply with the independent screening of AβO using a combination of the thioflavin-T and X-34 fluorophores. Altogether, our findings illustrate that different S100B multimers act as complementary suppressors of Aβ42 oligomerization and aggregation, further underpinning their potential neuroprotective role in AD

Human gingival fibroblasts response to different endodontic sealers: an in vitro study

Endodontic treatment aims to eliminate infection of the root canals and fill the dental pulp space. The biocompatibility studies of the sealers used in root canals obturation are crucial since they are applied in direct contact with periradicular tissues. Objective: The aim of this study was to evaluate the cytotoxicity of three root canal sealers—AH Plus, Bio MTA+, and Bio C sealer—on immortalized human gingival fibroblasts. Methods: AH Plus, Bio MTA+, and Bio C sealers were evaluated through incubation in real-time and material-conditioned media. Cells were incubated for 24 h and 72 h, at three different concentrations (1, 10, and 100 mg/mL) of each sealer. The cytotoxic activity of the sealers was assessed by Methyl tetrazolium (MTT) and Sulforhodamine B (SRB) assays. Cell morphology and cytogenetic alterations were studied microscopically. Results: MTT and SRB assays revealed similar results within both approaches. Cell culture exposed to sealers through incubation in real-time revealed a cytotoxic effect of AH Plus at 100 mg/mL. Material- conditioned media study revealed a cytotoxic effect of Bio MTA+ and Bio C, increasing with higher compound concentration and reaching 50% with 100 mg/mL. Regarding the cell’s morphology, Bio C sealer revealed a decrease in cell confluence and several morphological changes. AH Plus and Bio MTA+ did not seem to affect the cell confluence however morphology alterations were observed. In the cytogenetic study, a severe decrease of the mitotic index and a large number of chromosomal aberrations were observed. The present study represents an advance in the understanding of the biocompatibility of AH Plus, Bio MTA+, and Bio C sealers. These sealers demonstrated some cytotoxicity, depending on the concentration used. Although more validation studies are still needed, this study brings very relevant results in terms of cytotoxicity, cell morphology, and cytogenetic alterations. Conclusions: These results could help in the selection of the most appropriate compounds to be used in clinical practice as well as to determine the maximum recommended amounts of each sealer. Clinical Relevance: This study highlights the potential cytotoxic effects of three commonly used root canal sealers on human gingival fibroblasts, with varying degrees of impact depending on the concentration used. The results emphasize the importance of careful consideration when selecting and applying these materials in clinical practice.info:eu-repo/semantics/publishedVersio

Characterization of phenolic compounds of OMW : toxicity and degradability by yeasts

The characterization of Olive Mill Wastewaters (OMW), focusing the phenolic compounds, is one of the aims of the present work. As a first approach to characterize the phenolic compounds of OMW, the extraction methods used were: a liquid-liquid extraction by acidified ethyl acetate and a solid-liquid extraction with acidified methanol. The analysis of these extracts by reversed phase liquid chromatography confirmed that hydroxytyrosol was the most abundant phenolic compound in OMW, and that this compound was more efficiently recovered by the solid-liquid extraction technique. It was also a goal of this work to study the phenolic compounds toxicity to some yeast strains. Among the phenolic compounds tested catechol is the most inhibitory one to the cells. The phenols degradation was quite difficult, particularly when more easily degradable carbon source is still present in the medium.Fundação para a Ciência e a Tecnologia (FCT) - PTDC/AMB/69379/2006, SFRH/BD/27915/200

Flour fortification for nutritional and health improvement: A review

Deficiencies of micronutrients, essentially vitamins and minerals, have mainly cause several illnesses, especially in children and women worldwide. Governments and world organizations have focused great efforts to address these shortcomings and improve the health of the populations. This malnutrition can be combated by fortifying basic foods that are consumed daily. Thus, flours, especially wheat, maize and rice, are a popular and common food in the world's populations diet and are gaining considerable attention as a suitable vehicle for micronutrient fortification. There are some studies that show the effectiveness of flour fortification in combating micronutrient deficiencies and several diseases and the flour fortification strategies are directed to meet nutritional and health needs of the populations. The main points discussed in this review are food fortification, with great focus in flour fortification, health benefits, and legislative issues. This review also highlights multifaceted issues related to flour fortification to meet nutritional needs and to improve the health of vulnerable populations.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019), R.V.C. Cardoso grant (SFRH/BD/137436/2018). L. Barros and A. Fernandes also thank the national funding by FCT – Foundation for Science and Technology, P.I., through the institutional scientific employment program-contract for their contracts. This work is funded by the European Structural and Investment Funds (FEEI) through the Regional Operational Program North 2020, within the scope of Project Mobilizador ValorNatural®; and to FEEI through the Rural Development Program (PDR2020).info:eu-repo/semantics/publishedVersio

NIVEIS DE SUPERÓXIDO DISMUTASE REVELAM STRESS OXIDATIVO AUMENTADO NA POLINEUROPATIA AMILOIDÓTICA FAMILIAR

NIVEIS DE SUPERÓXIDO DISMUTASE REVELAM STRESS OXIDATIVO AUMENTADO NA POLINEUROPATIA AMILOIDÓTICA FAMILIAR Henrique Reguengo1,2, Maria Luís Cardoso2, Teresa Coelho3, Ana Martins3, Marta Novais3, Madalena Cruz1, Isabel Fonseca3, Berta Martins4, Franklim Marques2 1Serviço de Química Clínica, HSA/CHP, 2FFUP, 3Unidade Clínica de Paramiloidose, HSA/CHP, 4Laboratório de Imunogenética, ICBAS/UP. Hospital de Santo António, Centro Hospitalar do Porto (HSA/CHP), Porto. Faculdade de Farmácia, Universidade do Porto (FF/UP), Porto. Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto (ICBAS/UP), Porto. Introdução A polineuropatia amiloidótica familiar (PAF) é provocada por mutações no gene da transtirretina, principalmente em resultado da mutação TTRv30M no caso de Portugal. Estudos recentes em vários tipos de amiloidoses revelam que o stress oxidativo pode estar envolvido quer na produção das fibrilas de proteína amilóide, quer na modificações pós formação das fibrilas. A enzima superóxido dismutase (SOD) desempenha um importante papel antioxidante que protege as células expostas aos radicais superóxido. O presente estudo pretendeu avaliar o stress oxidativo nesta patologia. Material e Métodos A amostra em estudo incluiu 40 doentes com PAF e 45 portadores assintomáticos da mutação, seguidos habitualmente na Unidade Clínica de Paramiloidose do CHP. Foi ainda considerado um grupo controlo de 26 indivíduos saudáveis. Avaliou-se a concentração de SOD intra-eritocitária com um método colorimétrico disponível no KIT RANSOD ref SD 125 da Randox. A análise estatística foi efectuada utilizando o software SPSS, versão 19. Resultados Os valores de SOD obtidos foram os seguintes: Controlos: (1208±254 U/g Hb), Portadores assintomáticos (1436±62 U/g Hb), Doentes com PAF (1455±350 U/g Hb). Os valores resultaram da média de três réplicas. Os valores de SOD foram significativamente mais elevados no grupo de doentes com PAF e nos portadores assintomáticos, comparativamente ao grupo controlo (respectivamente P=0.003 e P=0.013). Não foram encontradas diferenças estatisticamente significativas entre os doentes com PAF e os portadores assintomáticos. No grupo de doentes com PAF verificou-se uma correlação positiva significativa entre os valores de SOD e Proteína C reactiva (r=0.45, P =0.013). Conclusão A alteração da concentração da SOD nos doentes com PAF sugere uma maior exposição dos mesmos a fenómenos de stress oxidativo comparativamente ao grupo controlo. Dado que este fenómeno pode ter influência quer no despoletar, quer no curso da patologia da doença, justifica-se uma maior atenção e estudo deste fenómeno nestes doentes. Apresentador: Henrique Reguengo, Técnico Superior de Saúde, Serviço de Quimica Clínica, HSA/CHP; Aluno de Doutoramento em Ciências Farmacêuticas, FF/UP

Characterization of phenolic compounds of OMW: toxicity and degrability by yeasts

The olive oil extraction, performed by the 3-phases process, results on a large amount of an effluent, usually known as Olive Mill Wastewater (OMW). It is mainly produced in the Mediterranean area, with the exception of Spain that is one of the major producers but mostly use the 2-phases process. OMW represents a major environmental problem due to its high organic content, being characterized by a strong acidic smell and an intensive brown to dark color due to the presence of biodegradable, recalcitrant and biostatic compounds. OMW phytotoxicity can be attributed to the phenolic compounds (Lanciotti et al, 2005). Owing to their instability, OMW phenols tend to polymerise during storage into condensed high-molecular-weight polymers that are difficult to degrade (Crognale et al, 2006). Thus, uncontrolled OMW disposal can create severe risks to water and soil quality. OMW is currently concentrated by evaporation in open pools, but this method is not satisfactory because a black foul-smelling sludge, difficult to remove, is produced. Instead of disposal solutions an approach of using this waste as a resource to be valorized is of greater interest. In fact, OMW contains sugars, lipids, mineral elements and phenolic compounds (10 % of the organic matter) that could be either directly recovered by chemical extraction and subsequent purification, or utilized as a basis for fermentative processes

Patterned piezoelectric scaffolds for osteogenic differentiation

The morphological clues of scaffolds can determine cell behavior and, therefore, the patterning of electroactive polymers can be a suitable strategy for bone tissue engineering. In this way, this work reports on the influence of poly(vinylidene fluoride-co-trifluoroethylene) (P(VDF-TrFE)) electroactive micropatterned scaffolds on the proliferation and differentiation of bone cells. For that, micropatterned P(VDF-TrFE) scaffolds were produced by lithography in the form of arrays of lines and hexagons and then tested for cell proliferation and differentiation of pre-osteoblast cell line. Results show that more anisotropic surface microstructures promote bone differentiation without the need of further biochemical stimulation. Thus, the combination of specific patterns with the inherent electroactivity of materials provides a promising platform for bone regeneration.This work was supported by national funds through the Fundação para a Ciência e Tecnologia (FCT) and by ERDF through COMPETE2020—Programa Operacional Competitividade e Internacionalização (POCI) in the framework of the Strategic Programs UID/FIS/04650/2020 and UIDB/04436/2020 and projects PTDC/EMD-EMD/28159/2017 and PTDC/BTM-MAT/28237/2017. TA thank FCT for the grant SFRH/BD/141136/2018 and CR thanks the FCT for the contract under the Stimulus of Scientific Employment (DL57/2016 junior researcher contract). Finally, the authors acknowledge funding by Spanish State Research Agency (AEI) and the European Regional Development Fund (ERFD) through the project PID2019-106099RB-C43/AEI/10.13039/501100011033 and from the Basque Government Industry and Education Departments under the ELKARTEK and PIBA (PIBA-2018-06) programs, respectively

MicroRNA deregulation and chemotaxis and phagocytosis impairment in Alzheimer's disease

AbstractIntroductionMononuclear phagocytes play a critical role during Alzheimer's disease (AD) pathogenesis due to their contribution to innate immune responses and amyloid beta (Aβ) clearance mechanisms.MethodsBlood-derived monocytes (BDMs) and monocyte-derived macrophages (MDMs) were isolated from blood of AD, mild cognitive impairment (MCI) patients, and age-matched healthy controls for molecular and phenotypic comparisons.ResultsThe chemokine/chemokine receptor CCL2/CCR2 axis was impaired in BDMs from AD and MCI patients, causing a deficit in cell migration. Changes were also observed in MDM-mediated phagocytosis of Aβ fibrils, correlating with alterations in the expression and processing of the triggering receptor expressed on myeloid cells 2 (TREM2). Finally, immune-related microRNAs (miRNAs), including miR-155, -154, -200b, -27b, and -128, were found to be differentially expressed in these cells.DiscussionThis work provides evidence that chemotaxis and phagocytosis, two crucial innate immune functions, are impaired in AD and MCI patients. Correlations with miRNA levels suggest an epigenetic contribution to systemic immune dysfunction in AD