Assessing chronological aging in Saccharomyces cerevisiae.

Chronological age represents the time that passes between birth and a given date. To understand the complex network of factors contributing to chronological lifespan, a variety of model organisms have been implemented. One of the best studied organisms is the yeast Saccharomyces cerevisiae, which has greatly contributed toward identifying conserved biological mechanisms that act on longevity. Here, we discuss high- und low-throughput protocols to monitor and characterize chronological lifespan and chronological aging-associated cell death in S. cerevisiae. Included are propidium iodide staining with the possibility to quantitatively assess aging-associated cell death via flow cytometry or qualitative assessments via microscopy, cell viability assessment through plating and cell counting and cell death characterization via propidium iodide/AnnexinV staining and subsequent flow cytometric analysis or microscopy. Importantly, all of these methods combined give a clear picture of the chronological lifespan under different conditions or genetic backgrounds and represent a starting point for pharmacological or genetic interventions.

Neonatal exposure to hypoxia induces early arterial stiffening via activation of lysyl oxidases.

Hypoxia in the neonatal period is associated with early manifestations of adverse cardiovascular health in adulthood including higher risk of hypertension and atherosclerosis. We hypothesize that this occurs due to activation of lysyl oxidases (LOXs) and the remodeling of the large conduit vessels, leading to early arterial stiffening. Newborn C57Bl/6 mice were exposed to hypoxia (FiO2  = 11.5%) from postnatal day 1 (P1) to postnatal day 11 (P11), followed by resumption of normoxia. Controls were maintained in normoxia. Using in vivo (pulse wave velocity; PWV) and ex vivo (tensile testing) arterial stiffness indexes, we determined that mice exposed to neonatal hypoxia had significantly higher arterial stiffness compared with normoxia controls by young adulthood (P60), and it increased further by P120. Echocardiography performed at P60 showed that mice exposed to hypoxia displayed a compensated dilated cardiomyopathy. Western blotting revelated that neonatal hypoxia accelerated age-related increase in LOXL2 protein expression in the aorta and elevated LOXL2 expression in the PA at P11 with a delayed decay toward normoxic controls. In the heart and lung, gene and protein expression of LOX/LOXL2 were upregulated at P11, with a delayed decay when compared to normoxic controls. Neonatal hypoxia results in a significant increase in arterial stiffness in early adulthood due to aberrant LOX/LOXL2 expression. This suggests an acceleration in the mechanical decline of the cardiovascular system, that contributes to increased risk of hypertension in young adults exposed to neonatal hypoxia that may increase susceptibility to further insults.

A 24-month clinical evaluation of composite resins with different viscosity and chemical compositions: a randomized clinical trial.

OBJECTIVES: To evaluate the clinical performance of two methacrylate-based flowable composites and an ormocer-based flowable composite in noncarious cervical lesions (NCCLs) in adult participants. METHOD AND MATERIALS: In total, 183 restorations were performed on NCCLs. All cavities were restored using a universal adhesive system (Futurabond U, Voco) with selective enamel etching and with one of the three evaluated flowable composites (n = 61): low-viscosity methacrylate-based composite (GrandioSO Flow, LV), high-viscosity methacrylate-based composite (GrandioSO Heavy Flow, HV), and an ormocer-based flowable composite (Admira Fusion Flow, ORM). All restorations were evaluated using FDI and USPHS criteria after 24 months. Kruskall-Wallis analysis of variance rank (α = .05) was used for statistical analysis. RESULTS: After 24 months of clinical evaluation, 16 restorations were lost (LV = 3, HV = 10, ORM = 3) and the retention rates (95% confidence interval) were 95.0% for LV, 82.2% for HV, and 95.0% for ORM, with statistical differences observed between HV and LV as well as HV and ORM (P < .05). When secondary parameters were evaluated, no significant differences between groups were observed (P > .05). Thirty-three restorations (LV = 8, HV = 13, ORM = 12) showed minor marginal staining, 71 restorations (LV = 26, HV = 20, ORM = 25) presented small marginal adaptation defects, and one restoration for HV presented recurrence of caries. CONCLUSION: The universal adhesive associated with the ormocer-based and methacrylate-based flowable composite showed promising clinical performance after 24 months. However, the heavy-flow restorations showed significantly more failures. (Quintessence Int 2023;54:186-199; doi: 10.3290/j.qi.b3631841).

Redescription of Leptoconops (Leptoconops) brasiliensis (Lutz) and Leptoconops (Megaconops) floridensis Wirth (Diptera: Ceratopogonidae) with its first Brazilian record.

The Leptoconops Skuse genus includes many pestiferous biting midges found in coastal areas. Only two species were previously found in Brazil and some species have ancient descriptions lacking updated taxonomic features. Herein, we redescribe Leptoconops (Leptoconops) brasiliensis Lutz and L. (Megaconops) floridensis Wirth based on female type specimens and additional material from Gois and Santa Catarina, Brazil. The first Brazilian record of L. (M.) floridensis is reported.

Dinámica de la evolución de las formas de Blastocystis spp. en un medio de cultivo simple / Dynamics of the evolution of the forms of Blastocystis spp. in a simple culture medium

Resumen Blastocystis spp. es un parásito muy frecuente en materia fecal humana, pero la naturaleza polimórfica y el número de Blastocystis en la muestra pueden complicar su detección por microscopía. El objetivo del trabajo fue describir la dinámica de los morfotipos de Blastocystis a corto plazo en un medio de cultivo simple y determinar su aplicabilidad para utilizarlo como complemento del análisis coproparasitológico y para estudios morfológicos, bioquímicos y moleculares del parásito. Se sembraron 10 muestras de materia fecal con Blastocystis en un medio Pavlova adaptado, se examinaron diariamente por examen microscópico durante 6 días y se registraron las formas y el recuento. El desarrollo fue regular y abundante y las formas fueron de tamaños variables y claramente identificables. El cultivo ensayado puede ser útil para la detección de Blastocystis cuando existan dudas diagnósticas por microscopía, para estudios de sensibilidad y especificidad diagnóstica o cuando se requiera aumentar la carga para realizar otros estudios.

Abstract Blastocystis spp. is a very frequent parasite in human fecal matter, but the polymorphic nature and the number of Blastocystis in a sample can complicate its detection by microscopy. The objective of the present work was to describe the dynamics of Blastocystis morphotypes in the short term in a simple culture medium and to determine its applicability to use it as a complement to coproparasitological analysis and for morphological, biochemical and molecular studies of the parasite. Ten stool samples with Blastocystis were cultured in an adapted Pavlova medium and examined during 6 days by microscopy to record the forms and the count. The development was regular and abundant and the shapes were of variable sizes and clearly identifiable. The tested culture could be used for the detection of Blastocystis when microscopic diagnosis is dubious, for studies of diagnostic sensitivity and specificity or when it is necessary to increase the load to perform other studies.

Resumo Blastocystis spp. é um parasita muito frequente nas fezes humanas, mas a natureza polimórfica e o número de Blastocystis na amostra podem complicar a sua detecção através do microscópio. O objetivo do trabalho foi descrever a dinâmica dos morfotipos de Blastocystis no curto prazo em um meio de cultura simples e determinar sua aplicabilidade para ser utilizado como complemento da análise coproparasitológica e para estudos morfológicos, bioquímicos e moleculares do parasita. Foram semeadas dez amostras de fezes com Blastocystis em um meio Pavlova adaptado e examinadas diariamente através de exame microscópico durante 6 dias, registrando as formas e fazendo recontagem. O desenvolvimento foi regular e abundante e as formas foram de tamanhos variáveis e claramente identificáveis. A cultura testada pode ser útil para a detecção de Blastocystis quando houver dúvidas diagnósticas por microscopia; para estudos de sensibilidade e especificidade diagnóstica ou quando for necessário aumentar a carga para a realização de outros estudos.

The rise of Candida auris: from unique traits to co-infection potential.

Candida auris is a multidrug resistant (MDR) fungal pathogen with a crude mortality rate of 30-60%. First identified in 2009, C. auris has been rapidly emerging to become a global risk in clinical settings and was declared an urgent health threat by the Centers for Disease Control and Prevention (CDC). A concerted global action is thus needed to successfully tackle the challenges created by this emerging fungal pathogen. In this brief article, we underline the importance of unique virulence traits,including its easy transformation, its persistence outside the host and its resilience against multiple cellular stresses, as well as of environmental factors that have mainly contributed to the rise of this superbug.

Selected Transesophageal Echocardiographic Parameters of Left Ventricular Diastolic Function Predict Length of Stay Following Coronary Artery Bypass Graft-A Prospective Observational Study.

(1) Importance: Abnormal left ventricular (LV) diastolic function, with or without a diagnosis of heart failure, is a common finding that can be easily diagnosed by intra-operative transesophageal echocardiography (TEE). The association of diastolic function with duration of hospital stay after coronary artery bypass (CAB) is unknown. (2) Objective: To determine if selected TEE parameters of diastolic dysfunction are associated with length of hospital stay after coronary artery bypass surgery (CAB). (3) Design: Prospective observational study. (4) Setting: A single tertiary academic medical center. (5) Participants: Patients with normal systolic function undergoing isolated CAB from September 2017 through June 2018. (6) Exposures: LV function during diastole, as assessed by intra-operative TEE prior to coronary revascularization. (7) Main Outcomes and Measures: The primary outcome was duration of postoperative hospital stay. Secondary intermediate outcomes included common postoperative cardiac, respiratory, and renal complications. (8) Results: The study included 176 participants (mean age 65.2 ± 9.2 years, 73% male); 105 (60.2%) had LV diastolic dysfunction based on selected TEE parameters. Median time to hospital discharge was significantly longer for subjects with selected parameters of diastolic dysfunction (9.1/IQR 6.6−13.5 days) than those with normal LV diastolic function (6.5/IAR 5.3−9.7 days) (p < 0.001). The probability of hospital discharge was 34% lower (HR 0.66/95% CI 0.47−0.93) for subjects with diastolic dysfunction based on selected TEE parameters, independent of potential confounders, including a baseline diagnosis of heart failure. There was a dose−response relation between severity of diastolic dysfunction and probability of discharge. LV diastolic dysfunction based on those selected TEE parameters was also associated with postoperative cardio-respiratory complications; however, these complications did not fully account for the relation between LV diastolic dysfunction and prolonged length of hospital stay. (9) Conclusions and Relevance: In patients with normal systolic function undergoing CAB, diastolic dysfunction based on selected TEE parameters is associated with prolonged duration of postoperative hospital stay. This association cannot be explained by baseline comorbidities or common post-operative complications. The diagnosis of diastolic dysfunction can be made by TEE.

A hundred spotlights on microbiology: how microorganisms shape our lives.

Viral, bacterial, fungal and protozoal biology is of cardinal importance for the evolutionary history of life, ecology, biotechnology and infectious diseases. Various microbiological model systems have fundamentally contributed to the understanding of molecular and cellular processes, including the cell cycle, cell death, mitochondrial biogenesis, vesicular fusion and autophagy, among many others. Microbial interactions within the environment have profound effects on many fields of biology, from ecological diversity to the highly complex and multifaceted impact of the microbiome on human health. Also, biotechnological innovation and corresponding industrial operations strongly depend on microbial engineering. With this wide range of impact in mind, the peer-reviewed and open access journal Microbial Cell was founded in 2014 and celebrates its 100th issue this month. Here, we briefly summarize how the vast diversity of microbiological subjects influences our personal and societal lives and shortly review the milestones achieved by Microbial Cell during the last years.

New species and new records of Monohelea Kieffer (Diptera, Ceratopogonidae) from Brazil.

Two new Brazilian species of Monohelea Kieffer are described and illustrated based on male specimens, Monoheleacapixaba sp. nov. from Espírito Santo and Monoheleacoimbrai sp. nov. from Rio de Janeiro. New records for M.aguirrei Tavares & Souza, M.archibaldoi Tavares & Souza and M.maculipennis (Coquillet) are given based on specimens from Espírito Santo (all three species) and Amapá (M.maculipennis only). All specimens are deposited in the Ceratopogonidae Collection of Fundação Oswaldo Cruz, Brazil.

Murals meet microbes: at the crossroads of microbiology and cultural heritage.

Our cultural heritage consists of manifold cultural expressions and represents a defining feature of our societies that needs to be further inherited to future generations. Even though humankind always fought a daily struggle for survival, at the same time, it seemed to have a spiritual need that went far beyond mere materialistic satisfaction and nowadays manifests in sometimes very ancient, yet brilliant artistic works. This fundamental legacy is endangered by several instances, including biodeterioration. Indeed, microorganisms play a significant role in the decline of all forms of tangible cultural heritage, including movable, immovable and underwater cultural heritage. Microbial colonization, biofilm formation and damaging metabolite production eventually result in critical decay. Thus, efforts to mitigate the negative impact of damaging microorganisms have been pursued with diverse physical, chemical and biological approaches. Intriguingly, recent advances have unveiled that specific microorganisms and microbial-based technologies also have the potential for cultural heritage preservation and present unique advantages. This short piece provides a quick overview on the duality of microorganisms in the conservation and restoration of cultural heritage.

Probing tissue transglutaminase mediated vascular smooth muscle cell aging using a novel transamidation-deficient Tgm2-C277S mouse model.

Tissue transglutaminase (TG2), a multifunctional protein of the transglutaminase family, has putative transamidation-independent functions in aging-associated vascular stiffening and dysfunction. Developing preclinical models will be critical to fully understand the physiologic relevance of TG2's transamidation-independent activity and to identify the specific function of TG2 for therapeutic targeting. Therefore, in this study, we harnessed CRISPR-Cas9 gene editing technology to introduce a mutation at cysteine 277 in the active site of the mouse Tgm2 gene. Heterozygous and homozygous Tgm2-C277S mice were phenotypically normal and were born at the expected Mendelian frequency. TG2 protein was ubiquitously expressed in the Tgm2-C277S mice at levels similar to those of wild-type (WT) mice. In the Tgm2-C277S mice, TG2 transglutaminase function was successfully obliterated, but the transamidation-independent functions ascribed to GTP, fibronectin, and integrin binding were preserved. In vitro, a remodeling stimulus led to the significant loss of vascular compliance in WT mice, but not in the Tgm2-C277S or TG2-/- mice. Vascular stiffness increased with age in WT mice, as measured by pulse-wave velocity and tensile testing. Tgm2-C277S mice were protected from age-associated vascular stiffening, and TG2 knockout yielded further protection. Together, these studies show that TG2 contributes significantly to overall vascular modulus and vasoreactivity independent of its transamidation function, but that transamidation activity is a significant cause of vascular matrix stiffening during aging. Finally, the Tgm2-C277S mice can be used for in vivo studies to explore the transamidation-independent roles of TG2 in physiology and pathophysiology.

Assessment of the effect of experimental bleaching agent with nano-bioactive material on postoperative sensitivity: A randomized, triple blind clinical trial.

OBJECTIVES: This clinical study aimed to evaluate the effect of incorporating bioactive nanoparticles (n-Bm) inside an in-office bleaching gel on the risk and intensity of tooth sensitivity (TS) and on bleaching effectiveness. MATERIALS AND METHODS: Sixty-six participants were selected and randomly assigned into two groups: control-only in-office gel and experimental-in-office gel with n-Bm. Teeth were bleached in two sessions (3 × 15-min). TS was recorded using a VAS and NRS. The color change was evaluated by subjective (VITA Classical and VITA Bleachedguide) and objective (Easyshade spectrophotometer) methods at baseline and 30 days after the end of treatment. The TS was evaluated by McNemar, Wilcoxon Signed Rank, and paired t test. The color changes between groups were compared using paired t test (α = 0.05). RESULTS: No significant differences between the groups were observed in the risk (control = 27% [95%IC 18-39]; experimental = 21% [95%IC 13-32]) and intensity of TS, as well as in the color change (p >0.05) for any color measurement. CONCLUSION: The inclusion of n-Bm into the bleaching agents did not affect the whitening effectiveness, as well as the risk and intensity of TS between groups. However, the results of the absolute risk of TS were low for both in-office gels used. CLINICAL SIGNIFICANCE: Despite no significant differences between groups, both experimental bleaching agents present suitable results with low values for TS.

Clinical performance of a micro invasive treatment based on demineralization-remineralization for dental fluorosis: case report

Introduction: Dental fluorosis manifests clinically as white to brown spots on the enamel teeth that were exposed to excessive fluoride during the formation process. Esthetic treatments were described in the literature to reduce or eliminate the fluorotic stains on the enamel surface. Objective: The aim of this study is to present an alternative treatment for fluorosis-stained teeth. Case report: This case report describes the clinical performance of a treatment of a 12-year-old male patient whose teeth presented moderate fluorosis. This treatment is based on tooth demineralization and remineralization. The material has an acid phase made by hydrochloric acid with tricarboxylic acid, and an alkaline phase made by Calcium Hydroxide. Results: This pain less and fast treatment presented good results. The treatment eliminated the spots during the follow-up and preserved most of the dental structure, improving the appearance of the patient's teeth. Conclusion: The appearance of the treated enamel showed a surface almost completely free of fluorotic stains, demonstrating the satisfactory results of this treatment.

Introdução: A fluorose dentária manifesta-se clinicamente como manchas brancas a marrons no esmalte de dentes expostos ao excesso de flúor durante o processo de formação. Tratamentos estéticos foram descritos na literatura para reduzir ou eliminar as manchas fluoróticas na superfície do esmalte. Objetivo: Apresentar uma alternativa de tratamento para dentes manchados por fluorose dentária. Relato do caso: Este relato de caso descreve o desempenho clínico do tratamento para dentes com fluorose moderada em um paciente de 12 anos. Esse tratamento foi baseado numa técnica de desmineralização e remineralização do dente. O material possui uma fase ácida composta por ácido clorídrico com ácido tricarboxílico, e uma fase alcalina composta por Hidróxido de Cálcio. Resultados: Observou-se que esse tratamento, sem dor e rápido, apresentou resultados satisfatórios, pois eliminou as manchas durante o acompanhamento. Além disso, preservou ao máximo a estrutura dentária, beneficiando o paciente com uma melhor aparência dos seus dentes. Conclusão: O aspecto do esmalte tratado mostrou uma superfície quase sem manchas fluoróticas, demonstrando resultados satisfatórios deste tratamento.

The role of HIF-1α in nicotine-induced root and bone resorption during orthodontic tooth movement.

BACKGROUND: In orthodontic tooth movement (OTM), pseudo-inflammatory processes occur that are similar to those of nicotine-induced periodontitis. Previous studies have shown that nicotine accelerates OTM, but induces periodontal bone loss and dental root resorption via synergistically increased osteoclastogenesis. This study aimed to investigate the role of hypoxia-inducible factor 1 alpha (HIF-1α) in nicotine-induced osteoclastogenesis during OTM. MATERIALS/METHODS: Male Fischer-344 rats were treated with l-Nicotine (1.89 mg/kg/day s.c., N = 10) or NaCl solution (N = 10). After a week of premedication, a NiTi spring was inserted to mesialize the first upper left molar. The extent of dental root resorption, osteoclastogenesis, and HIF-1α protein expression was determined by (immuno)histology, as well as bone volume (BV/TV) and trabecular thickness (TbTh) using µCT. Receptor activator of nuclear factor of activated B-cells ligand (RANK-L), osteoprotegerin (OPG), and HIF-1α expression were examined at the protein level in periodontal ligament fibroblasts (PDLF) exposed to pressure, nicotine and/or hypoxia, as well as PDLF-induced osteoclastogenesis in co-culture experiments with osteoclast progenitor cells. RESULTS: Nicotine favoured dental root resorptions and osteoclastogenesis during OTM, while BV/TV and TbTh were only influenced by force. This nicotine-induced increase does not appear to be mediated by HIF-1α, since HIF-1α was stabilized by force application and hypoxia, but not by nicotine. The in vitro data showed that the hypoxia-induced increase in RANK-L/OPG expression ratio and PDLF-mediated osteoclastogenesis was less pronounced than the nicotine-induced increase. CONCLUSIONS: Study results indicate that the nicotine-induced increase in osteoclastogenesis and periodontal bone resorption during OTM may not be mediated by hypoxic effects or HIF-1α stabilization in the context of nicotine-induced vasoconstriction, but rather by an alternative mechanism.

Influence of viscosity and chemical composition of composite resins in non-carious cervical restorations: 12-month randomized clinical trial / Influência da viscosidade e composição química das resina compostas em restaurações de lesões cervicais não-cariosas: resultados de 12 meses de uma pesquisa clínica randomizada

Objective: The objective of this double-blind, randomized controlled clinical trial was to evaluate the clinical performance of two methacrylate-based flowable composite and ormocer-based flowable composite in non-carious cervical lesions (NCCLs) of adult patients. Material and Methods: 183 restorations were performed on NCCLs using the Futurabond U adhesive system, applied in the selective enamel etching mode in all cavities. After the adhesive application, the cavities were restored with one out of the three evaluated flowable composites (n = 61 per group): ormocer-based flowable composite (Admira Fusion Flow, ORM), low viscosity methacrylate-based composite (GrandioSO Flow, LV) and high viscosity methacrylate-based composite (GrandioSO Heavy Flow, HV). After 12 months of clinical performance, these restorations were evaluated according to FDI and USPHS criteria in the following items: retention/fracture, marginal adaptation, marginal staining, postoperative sensitivity and caries recurrence. Results: eight restorations were lost/fractured after 12 months of clinical evaluation (1 in the ORM and 7 in the HV group). The retention rates for 12- months (95% confidence interval) were 98.4% (91.3%-99.7%) for the ORM group, 100% (94.5%-100%) for the LV group and 88.5% (78.1%-94.3%) for the HV group, with no statistical difference identified between any pair of groups (p > 0.05). Five restorations presented small marginal adaptation defects at the 12-months evaluation recall, and all of them were considered clinically acceptable. Conclusion: The clinical performance of the universal adhesive associated to ormocer-based or methacrylate-based flowable composite were found to be promising after 12-month of clinical evaluation (AU)

Objetivo: O objetivo deste estudo clínico duplo-cego randomizado foi comparar as taxas de retenção de um compósito fluido à base de Ormocer versus dois compósitos fluidos à base de metacrilato quando utilizados em lesões cervicais não cariosas (LCNCs) de pacientes adultos. Material e Métodos: 183 restaurações foram realizadas em LCNCs utilizando o sistema adesivo Futurabond U, aplicado no modo de condicionamento seletivo do esmalte em todas as cavidades. Após a aplicação do adesivo, as cavidades foram restauradas com um dos três compósitos fluidos avaliados (n = 61 por grupo): compósito fluido à base de ormocer (Admira Fusion Flow, ORM), compósito à base de metacrilato de baixa viscosidade (GrandioSO Flow, LV) e compósito à base de metacrilato de alta viscosidade (GrandioSO Heavy Flow, HV). Após 12 meses de desempenho clínico, essas restaurações foram avaliadas de acordo com os critérios FDI e USPHS nos seguintes itens: retenção / fratura, adaptação marginal, coloração marginal, sensibilidade pós-operatória e recorrência de cárie. Resultados: oito restaurações foram perdidas / fraturadas após 12 meses de avaliação clínica (1 no grupo ORM e 7 no grupo HV). As taxas de retenção por 12 meses (intervalo de confiança de 95%) foram 98,4% (91,3% -99,7%) para o grupo ORM, 100% (94,5% -100%) para o grupo LV e 88,5% (78,1% -94,3%) para o grupo HV, sem diferença estatística identificada entre nenhum par de grupos (p> 0,05). Cinco restaurações apresentaram pequenos defeitos de adaptação marginais no período de avaliação de 12 meses, e todas foram consideradas clinicamente aceitáveis. Conclusão: O desempenho clínico do adesivo universal associado ao compósito fluido à base de ormocer ou metacrilato mostrou-se promissor após 12 meses de avaliação clínica(AU)

Nutritional Aspects of Spermidine.

Natural polyamines (spermidine and spermine) are small, positively charged molecules that are ubiquitously found within organisms and cells. They exert numerous (intra)cellular functions and have been implicated to protect against several age-related diseases. Although polyamine levels decline in a complex age-dependent, tissue-, and cell type-specific manner, they are maintained in healthy nonagenarians and centenarians. Increased polyamine levels, including through enhanced dietary intake, have been consistently linked to improved health and reduced overall mortality. In preclinical models, dietary supplementation with spermidine prolongs life span and health span. In this review, we highlight salient aspects of nutritional polyamine intake and summarize the current knowledge of organismal and cellular uptake and distribution of dietary (and gastrointestinal) polyamines and their impact on human health. We further summarize clinical and epidemiological studies of dietary polyamines.

Comparative assessment of mouse models for experimental orthodontic tooth movement.

Animal experiments are essential for the elucidation of biological-cellular mechanisms in the context of orthodontic tooth movement (OTM). So far, however, no studies comparatively assess available mouse models regarding their suitability. OTM of first upper molars was induced in C57BL/6 mice either via an elastic band or a NiTi coil spring for three, seven or 12 days. We assessed appliance survival rate, OTM and periodontal bone loss (µCT), root resorptions, osteoclastogenesis (TRAP+ area) and local expression of OTM-related genes (RT-qPCR). Seven days after the elastic bands were inserted, 87% were still in situ, but only 27% after 12 days. Survival rate for the NiTi coil springs was 100% throughout, but 8.9% of the animals did not survive. Both methods induced significant OTM, which was highest after 12 (NiTi spring) and 7 days (band), with a corresponding increase in local gene expression of OTM-related genes and osteoclastogenesis. Periodontal bone loss and root resorptions were not induced at a relevant extent by neither of the two procedures within the experimental periods. To induce reliable OTM in mice beyond 7 days, a NiTi coil spring is the method of choice. The elastic band method is recommended only for short-term yes/no-questions regarding OTM.

Influence of flavonoids on long-term bonding stability on caries-affected dentin.

OBJECTIVES: To evaluate the effect of experimental dentin pre-treatment solutions formulated with different flavonoids on microtensile bond strength (µTBS), nanohardness (NH) and ultra-morphological characteristics of artificial caries-affected dentin (CAD) bonded using a universal bonding system. METHODS: A microbiological method was used to create an artificial CAD in 91 human molars. Five experimental pre-treatment solutions were created using the following flavonoids: quercetin (QUE); hesperidin (HES); rutin (RUT); naringin (NAR), or proanthocyanidin (PRO). A placebo solution (PLA) with no flavonoids added was also evaluated. The flavonoids or placebo solutions were applied to the CAD prior to the application and photoactivation of a universal adhesive (Scotchbond Universal, 3M Oral Care). A control group (CON), in which only the bonding agent was applied without any flavonoid solution, was also evaluated. A 3-mm-thick block of resin composite (Opallis, FGM) was built up on the flat bonded CAD surfaces and was light-cured following the manufacturer's instructions. Specimens were sectioned to obtain resin-dentin slices and sticks (cross-sectional area of 0.8 mm2). The µTBS, NH, and confocal ultramorphology analysis of resin-dentin interface was evaluated at 24 h and after thermo-cycling aging (25,000 cycles). The results were analyzed using 2-way ANOVA followed by Bonferroni's post hoc test (pre-set α = 0.05). RESULTS: The specimens from groups QUE, NAR, and RUT presented greater µTBS values than those from CON group (p<0.05). Specimens from some of these experimental groups presented greater nanomechanical properties (p<0.05), and no morphological degradation at the resin-dentin interface after aging. SIGNIFICANCE: The use of exogenous cross-linkers as dentin pre-treatment before bonding procedures may represent a suitable strategy to improve the longevity of universal adhesive systems applied to caries-affected dentin.

Fungal infections in humans: the silent crisis.

Annually, over 150 million severe cases of fungal infections occur worldwide, resulting in approximately 1.7 million deaths per year. Alarmingly, these numbers are continuously on the rise with a number of social and medical developments during the past decades that have abetted the spread of fungal infections. Additionally, the long-term therapeutic application and prophylactic use of antifungal drugs in high-risk patients have promoted the emergence of (multi)drug-resistant fungi, including the extremely virulent strain Candida auris. Hence, fungal infections are already a global threat that is becoming increasingly severe. In this article, we underline the importance of more and effective research to counteract fungal infections and their consequences.

Monumental architecture at Aguada Fénix and the rise of Maya civilization.

Archaeologists have traditionally thought that the development of Maya civilization was gradual, assuming that small villages began to emerge during the Middle Preclassic period (1000-350 BC; dates are calibrated throughout) along with the use of ceramics and the adoption of sedentism1. Recent finds of early ceremonial complexes are beginning to challenge this model. Here we describe an airborne lidar survey and excavations of the previously unknown site of Aguada Fénix (Tabasco, Mexico) with an artificial plateau, which measures 1,400 m in length and 10 to 15 m in height and has 9 causeways radiating out from it. We dated this construction to between 1000 and 800 BC using a Bayesian analysis of radiocarbon dates. To our knowledge, this is the oldest monumental construction ever found in the Maya area and the largest in the entire pre-Hispanic history of the region. Although the site exhibits some similarities to the earlier Olmec centre of San Lorenzo, the community of Aguada Fénix probably did not have marked social inequality comparable to that of San Lorenzo. Aguada Fénix and other ceremonial complexes of the same period suggest the importance of communal work in the initial development of Maya civilization.