ALS-associated VRK1 R321C mutation causes proteostatic imbalance and mitochondrial defects in iPSC-derived motor neurons.

Vaccinia-related kinase 1 (VRK1) is a gene which has been implicated in the pathological process of a broad range of neurodevelopmental disorders as well as neuropathies, such as Amyotrophic Lateral Sclerosis (ALS). Here we report a family presenting ALS in an autosomal recessive mode of inheritance, segregating with a homozygous missense mutation located in VRK1 gene (p.R321C; Arg321Cys). Proteomic analyses from iPSC-derived motor neurons identified 720 proteins eligible for subsequent investigation, and our exploration of protein profiles revealed significant enrichments in pathways such as mTOR signaling, E2F, MYC targets, DNA repair response, cell proliferation and energetic metabolism. Functional studies further validated such alterations, showing that affected motor neurons presented decreased levels of global protein output, ER stress and downregulation of mTOR signaling. Mitochondrial alterations also pointed to decreased reserve capacity and increased non-mitochondrial oxygen consumption. Taken together, our results present the main pathological alterations associated with VRK1 mutation in ALS.

Is tooth loss associated with oral health-related quality of life among young men? Findings from southern Brazil.

OBJECTIVE: To determine the association between tooth loss and oral health-related quality of life (OHRQoL) among young men from southern Brazil. METHODS: Cross-sectional study of 518 young men aged 18-19 years enlisted for mandatory military service in the Brazilian Army in two cities of southern Brazil in 2019-2021. Participants were clinically examined regarding gingivitis, dental caries experience (caries lesions, missing and filled surfaces), and malocclusion. A questionnaire collected socioeconomic information and the Oral Health Impact Profile 14 (OHIP-14). The outcome was OHRQoL, modeled as a discrete variable. The main explanatory variable was tooth loss due to caries (0 or ≥1). Negative binomial regression analysis was used; rate ratios (RR) and 95% confidence intervals (CI) were estimated. RESULTS: Tooth loss prevalence was 22.2%. Associations were detected between tooth loss and OHRQoL in the physical pain and handicap domains. Individuals with at least one missing tooth had 17% higher OHIP-14 scores in the physical pain domain (adjusted RR=1.17; 95%CI=1.01, 1.35) and 49% higher in the handicap domain (adjusted RR=1.49; 95%CI=1.03, 2.15) than those with no tooth loss. CONCLUSIONS: Tooth loss was negatively associated with OHRQoL among 18-19-year-old Brazilian Army conscripts, and this association was related to physical pain and handicap domains.

Sociodemographic disparities in oral health-related quality of life of schoolchildren in rural and urban areas.

OBJECTIVE: To assess the association between residence place, socioeconomic conditions and oral health-related quality of life (OHRQoL) among schoolchildren from southern Brazil. METHODS: Participants were 9-14-year-old schoolchildren from rural and urban municipal schools from Rosário do Sul, Brazil. The Child Perceptions Questionnaire (CPQ11-14) was used to assess OHRQoL. A structured questionnaire collected data on sociodemographic condition (family income), residence place (urban or rural), use of dental services, and behavioral variables. Clinical oral examination recorded the presence of missing teeth and the gingival bleeding index. Multilevel Poisson regression analysis with a hierarchical approach assessed the association between predictors and CPQ11-14 scores. Rate ratios (RR) and 95% confidence intervals (CI) were estimated. RESULTS: A total of 373 schoolchildren were included (rural area=122; urban area=251), with a mean CPQ11-14 score of 11.83, ranging from 0 to 42. Low-income schoolchildren living in rural areas had 15% higher CPQ11-14 scores than high-income schoolchildren living in urban areas. In urban areas, family income predicted OHRQoL, with low-income schoolchildren having 9% higher CPQ11-14 scores than high-income children. In rural areas, schoolchildren with low household income had 19% higher CPQ11-14 scores than high-income children. CONCLUSION: Schoolchildren from low-income families had a poorer OHRQoL irrespective of their area of residence (rural or urban). The association between family income and OHRQoL was more pronounced among children living in rural areas.

High prevalence of pathogenic Eimeria spp. and the main risk factors associated with infection in goats from a semiarid region of Northeastern Brazil.

This study aimed to analyze the epidemiology of infections caused by Eimeria spp. in goats bred in an extensive production system in Northeast Brazil. To this end, an epidemiological survey was performed, wherein 350 fecal samples were collected from goats ranging from 3 months to 8 years old across 30 farms. The number of oocysts per gram of feces (OPG) was counted using the Mini-FLOTAC method. Species were identified based on their morphological and morphometric characteristics. Frequencies between animal categories were assessed using the chi-square test and known risk factors after multivariate analysis. Eimeria oocysts were found in 99.4% (346/348) of samples and 100% (30/30) of the farms. The OPG was influenced by the age of the animals; therefore, young goats (up to 6 months old) eliminated more oocysts (2302.6 + 5454.8) than adult goats (881.7 + 1551.5). Three pathogenic species were identified, with two demonstrating a high prevalence: Eimeria arloingi (88.83%) and Eimeria christenseni (44.7%), in addition to Eimeria caprina (3.72%). In the study, the habit of ground feeding and the use of cisterns as water sources were identified as risk factors, with odds ratios of 4.04 and 9.82, respectively. However, cleaning facilities at shorter intervals was a protective factor (OR = 0.19). We concluded that there was a high prevalence of Eimeria infection in goats bred in extensive production systems, even those in semiarid regions. This high prevalence can be attributed to the congregation of animals in shaded areas and the substantial accumulation of fecal matter at night.

Deciphering the role of ttrA and pduA genes for Salmonella enterica serovars in a chicken infection model.

Salmonella enterica serovars use self-induced intestinal inflammation to increase electron acceptor availability and to obtain a growth advantage in the host gut. There is evidence suggesting that the ability of Salmonella to use tetrathionate and 1,2-propanediol provides an advantage in murine infection. Thus, we present here the first study to evaluate both systemic infection and faecal excretion in commercial poultry challenged by Salmonella Enteritidis (SE) and S. Typhimurium (STM) harbouring deletions in ttrA and pduA genes, which are crucial to the metabolism of tetrathionate and 1,2-propanediol, respectively. Mutant strains were excreted at higher rates when compared to the wild-type strains. The highest rates were observed with white egg-layer and brown egg-layer chicks (67.5%), and broiler chicks (56.7%) challenged by SEΔttrAΔpduA, and brown egg-layer chicks (64.8%) challenged by STMΔttrAΔpduA. SEΔttrAΔpduA presented higher bacterial counts in the liver and spleen of the three chicken lineages and caecal contents from the broiler chickens, whereas STMΔttrAΔpduA presented higher counts in the liver and spleen of the broiler and brown-egg chickens for 28 days post-infection (P < 0.05). The ttrA and pduA genes do not appear to be major virulence determinants in faecal excretion or invasiveness for SE and STM in chickens. RESEARCH HIGHLIGHTSttrA and pudA do not impair gut colonization or systemic infection in chicks.Mutant strains were present in higher numbers in broilers than in laying chicks.Mutants of SE and STM showed greater pathogenicity in broiler chicks than layers.

Ascaris lumbricoides eggs or artefacts? A diagnostic conundrum.

Due to the presence of artefacts in stool samples, the copromicroscopic diagnosis of Ascaris lumbricoides is not always straightforward, particularly in the case of fertilized decorticated eggs. A total of 286 stool samples from 115 schoolchildren in India and 171 adult immigrants in Italy were screened for the presence of A. lumbricoides eggs by both Kato-Katz thick smear and Mini-FLOTAC. If the outer layer of A. lumbricoides eggs was absent, two aliquots of each stool sample were preserved: one for coproculture to identify larvae after development and one to compose a pool of stool for molecular analysis. A total of 64 stool samples (22.4%) were positive for A. lumbricoides using the Kato-Katz thick smear; 36 (56.3%) of these showed mammillated A. lumbricoides eggs, 25 (39.1%) showed elements resembling fertilized decorticated eggs, while three samples (4.7%) showed both mammillated and decorticated eggs. By Mini-FLOTAC, 39 stool samples (13.6%) were positive, while decorticated A. lumbricoides-like eggs were identified as artefacts. These results were confirmed by negative coprocultures and quantitative polymerase chain reaction. Mini-FLOTAC can be used for a reliable diagnosis of A. lumbricoides, thanks to the flotation and translation features which allow a clearer view, resulting in the correct identification of A. lumbricoides eggs.

Antibacterial effects of the lectin from pomegranate sarcotesta (PgTeL) against Listeria monocytogenes.

AIMS: To investigate the effects of the lectin from Punica granatum sarcotesta (PgTeL) on growth, viability, cell structure, biofilm formation and chitinase activity of Listeria monocytogenes. In addition, the effect of PgTeL on the adhesion and invasion of human cells (HeLa) was determined. METHODS AND RESULTS: PgTeL showed bacteriostatic and bactericidal effects on the strains L. monocytogenes N53-1 and EGD-e, causing morphometric alterations, cell aggregation, strong deformation and cell disruption. PgTeL inhibited biofilm formation by EGD-e and N53-1 and also interfered with the adhesion and invasion processes of EGD-e and N53-1 in HeLa cells. Finally, the chitinase activity of L. monocytogenes EGD-e was reduced in the presence of PgTeL, which can be involved in the inhibition of adhesion process. CONCLUSION: PgTeL is an antibacterial agent against L. monocytogenes, inhibiting growth and promoting cell death, as well as impairing biofilm formation and bacterial adhesion and invasion into human cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The results stimulate future investigations on the potential of PgTeL for protection of contamination in food products.

Ultrastructural Aspects of The Intestinal Cells Surface of Angiostrongylus Cantonensis By Using of Transmission Electron Microscopy.

The rat lungworm Angiostrongylus cantonensis is a zoonotic parasite and the main cause of eosinophilic meningitis in the world. Its main source of nutrients originates from the degradation of host hemoglobin in blood-feeding helminths, such as A. cantonensis. The purpose of this study was to analyze the ultrastructure of the intestinal cells by using of transmission electron microscopy (TEM). A. cantonensis worms obtained from Rattus norvegicus (norway rats) from endemic area were used for TEM. The ultrastructural analysis was performed using fragments cut from the middle part of the worms, and the TEM study revealed the cells with microvilli and nuclei containing areas of loose and condensed chromatin and the presence of macrovesicles and microvesicles of digestion and it was observed in this study that intestinal epithelium is look like a syncytium. The ultrastructural aspects of the intestinal cells surface of A. Cantonensis the indicate that the intestinal epithelium is a multinucleate mosaic or syncytium.

Impact of markers of endothelial injury and hypercoagulability on systemic lupus erythematosus.

We have explored the relationship between possible hemostatic changes and clinical manifestation of the systemic lupus erythematosus (SLE) as a function of greater or lesser disease activity according to Systemic Lupus Erythematosus Disease Activity Index-2000 (SLEDAI-2K) criteria. Endothelial injury and hypercoagulability were investigated in patients with SLE by measuring thrombomodulin (TM), D-dimer (DDi) and thrombin generation (TG) potential. A total of 90 participants were distributed into three groups: 1) women with SLE presenting with low disease activity (laSLE) (SLEDAI-2K ≤ 4), 2) women with SLE presenting with moderate to high disease activity (mhaSLE) (SLEDAI-2K > 4), and 3) a control group comprising healthy women. Levels of TM and DDi were higher both in the laSLE and mhaSLE groups compared to controls and in mhaSLE compared to the laSLE group. With respect to TG assay, lagtime and endogen thrombin potential, low concentrations of tissue factor provided the best results for discrimination among groups. Analysis of these data allow us to conclude that TM, DDi and TG are potentially useful markers for discriminating patients with very active from those with lower active disease. Higher SLE activity may cause endothelial injury, resulting in higher TG and consequently a hypercoagulability state underlying the picture of thrombosis common in this inflammatory disease.

Long-term follow-up of inactive occlusal caries lesions: 4-5-year results.

OBJECTIVE: To assess the clinical behavior of inactive caries lesion on the occlusal sites of permanent molars over 4-5 years and to estimate the risk for progression of caries-inactive sites compared with sound ones. METHODS: Clinical examinations were conducted at baseline (n = 258) and after 4-5 years and included the recording of dental plaque and dental caries at the occlusal surfaces and the eruption stage of each permanent molar. RESULTS: One hudred ninety-three schoolchildren were followed (response rate of 74.8%), totalizing 1152 teeth. Of the children, 30.6% (n = 59) presented at least one molar containing an active lesion, filling, or that had been extracted; according to the activity criterion, inactive lesions presented around a twofold increased risk for caries progression than sound surfaces (OR = 2.34 95%CI = 1.51-3.62). Thirteen percent (n = 25) of the children presented at least one molar progressing to dentine cavity, filling, or extraction; according to the severity criterion, inactive caries lesions presented a significantly higher risk for progression when compared with sound surfaces (OR = 2.69, 95% CI = 1.50-4.83). CONCLUSION: The vast majority of lesions (85-90%) identified as inactive enamel caries at baseline did not progress over 4-5 years. Despite this fact, it was possible to detect an increased risk for caries progression in caries-inactive occlusal sites compared with the sound ones. CLINICAL RELEVANCE: Considering the low progression rates, inactive caries lesions do not need a specific caries-controlling treatment and should be monitored longitudinally in the same manner as sound surfaces.

Radiographic pattern of underlying dentin lesions (ICDAS 4) in permanent teeth.

OBJECTIVE: To assess the radiographic pattern of underlying dentine shadows (UDS) in the occlusal surfaces of permanent teeth. METHODS: A total of 282 permanent posterior teeth pertaining to 91 individuals, 142 UDS and 140 non-cavitated enamel lesions (NCEL), were included for comparison. UDS was defined as shadows of discolored dentin visible through the enamel surface which may or may not show signs of localized enamel breakdown, classified as code 4 by the International Caries Detection and Assessment System group. Data collection included the application of a questionnaire, clinical examination, and bilateral bitewing radiographs. The risk for presenting radiolucency was estimated using logistic regression model with generalized estimating equations. RESULTS: Approximately 79% of UDS exhibited no radiolucency. The proportion of teeth exhibiting a radiolucency restricted to the enamel-dentin junction was 20.4% for UDS and 3.6% for NCEL (p < 0.001, chi-square test). UDS had a sixfold increased risk for exhibiting radiolucency compared with NCEL (OR = 5.78, 95% CI = 2.73-12.22, p < 0.001). Despite this finding, it is important to highlight that virtually all cases were located at the enamel-dentin junction, and only one tooth in each category of clinical status exhibited radiolucency at the outer one half of dentin. No tooth exhibited radiolucency reaching the deep dentin. CONCLUSION: The present study showed that UDS presented radiolucency in very few cases. The vast majority of lesions showed no radiolucency. CLINICAL RELEVANCE: Our findings suggest that only a small proportion of UDS would demand restorative treatment.

Is there association between human development index and tuberculosis mortality risk? Evidence from a spatial analysis study in the south of Brazil.

The goal of this study was to analyse the spatial pattern of tuberculosis (TB) mortality using different approaches, namely: mortality rates (MR), spatial relative risks (RR) and Bayesian rates (Global and Local) and their association with human development index (HDI), Global and its three dimensions: education, longevity and income. An ecological study was developed in Curitiba, Brazil based on data from Mortality Information System (2008-2014). Spatial scan statistics were used to compute RR and identify high-risk clusters. Bivariate Local Indicator of Spatial Associations was used to assess associations. MR ranged between 0 and 25.24/100.000 with a mean (standard deviation) of 1.07 (2.66). Corresponding values for spatial RR were 0-27.46, 1.2 (2.99) and for Bayesian rates (Global and Local) were 0.49-1.66, 0.90 (0.19) and 0-6.59, 0.98 (0.80). High-risk clusters were identified for all variables, except for HDI-income and Global Bayesian rate. Significant negative spatial relations were found between MR and income; between RR and HDI global, longevity and income; and Bayesian rates with all variables. Some areas presented different patterns: low social development/low risk and high risk/high development. These results demonstrate that social development variables should be considered, in mortality due TB.

Action of natural phytosanitary products on Bacillus thuringiensis subsp. kurstaki S-1905.

The objective of this study was to evaluate the effects of natural phytosanitary products (NPs) on spores and crystals of Bacillus thuringiensis subsp. kurstaki S-1905 (Btk S-1905). For the spore assay, NPs and bacteria were applied in combination and individually. For the combined application, Btk S-1905 + NP mixtures were inoculated on nutrient agar (NA), and for the separate applications, the NPs were spread on NA plates, which were later inoculated with the pathogen. The number of colony-forming units (CFU) per milliliter was quantified after 18 h of incubation. For the crystal protein degradation assay, the Btk S-1905 + NP mixtures were added to the diet of Anticarsia gemmatalis (Lepidoptera: Erebidae), and mortality was evaluated at the following time points: 12, 24, 48, and 72 h. Scanning electron microscopy and agarose gel electrophoresis were carried out. Biogermex and Ecolife® reduced the CFU ml-1 in both combined and separate applications. Biogermex, Ecolife®, and Planta Clean were antagonistic to the action of bacterial toxins, and no product affected the morphology or resulted in the degradation of the crystal proteins. The remaining products evaluated did not reduce the CFU ml-1 and had additive effect when combined with the crystal toxin.

Partial caries removal in deep caries lesions: a 5-year multicenter randomized controlled trial.

OBJECTIVE: This multicenter randomized controlled clinical trial aimed to compare the outcomes of stepwise excavation (SW) and partial caries removal (PCR) regarding the maintenance of pulp vitality in deep caries lesions over 5 years. METHODS: At baseline, 299 permanent molars with deep caries lesions were randomly assigned to control or test groups. The control group received the stepwise excavation treatment (SW), while the test group received partial caries removal from the pulpal wall followed by restoration in a single session (PCR). Treatments were conducted in two centers located in the cities of Porto Alegre (South Brazil) and Brasília (Midwest Brazil). Survival analysis was performed to compare PCR and SW over time (Weibull regression models). The primary outcome of this study was pulp vitality, determined by the combination of the following characteristics: positive response to cold test, negative response to percussion, absence of spontaneous pain, and absence of periapical lesion (radiographic examination). RESULTS: This 5-year study includes data pertaining to 229 teeth: 121 teeth actually examined at the 5-year appointment, and 108 teeth contributed with data collected in previous follow-ups (18 months or 3 years). Survival analysis showed success rates of 80% in PCR group and 56% in SW group (p < 0.001). Failure was significantly associated with treatment [PCR, HR=0.38; 95%CI=0.23-0.63)] and region [South, HR=2.22; 95%CI=1.21-4.08]. CONCLUSION: PCR significantly reduced the occurrence of pulp necrosis when compared with SW. CLINICAL RELEVANCE: This study supports the PCR as a single-visit technique to manage deep caries lesions in permanent teeth.

Gas mixture for deep-UV plasma emission in a hollow-core photonic crystal fiber.

We report on the first deep-ultraviolet/ultraviolet (DUV/UV) emission using a highly compact microwave-driven plasma-core photonic crystal fiber. The latter consists of a few centimeter long micro-plasma column of a gas mixture in the core of Kagome hollow-core photonic crystal fiber. The plasma is generated by nonintrusively exciting a ternary gas mixture of argon, nitrogen, and oxygen (Ar/N2/O2) with a microwave resonator. Several spectral lines in the wavelength range of 200-450 nm were produced, guided by an Ar-N2-O2 plasma-filled fiber, and controlled by simply varying the gas ratio of this gas mixture. An optimum gas mixture ratio was experimentally and theoretically identified for the strongest emission in the DUV range of 200-275 nm. The developed DUV emitting plasma-core fiber represents an important milestone towards the development of tunable and miniaturized DUV/UV laser sources.

3D map distribution of metallic nanoparticles in whole cells using MeV ion microscopy.

In this work, a new tool was developed, the MORIA program that readily translates Rutherford backscattering spectrometry (RBS) output data into visual information, creating a display of the distribution of elements in a true three-dimensional (3D) environment. The program methodology is illustrated with the analysis of yeast Saccharomyces cerevisiae cells, exposed to copper oxide nanoparticles (CuO-NP) and HeLa cells in the presence of gold nanoparticles (Au-NP), using different beam species, energies and nuclear microscopy systems. Results demonstrate that for both cell types, the NP internalization can be clearly perceived. The 3D models of the distribution of CuO-NP in S. cerevisiae cells indicate the nonuniform distribution of NP in the cellular environment and a relevant confinement of CuO-NP to the cell wall. This suggests the impenetrability of certain cellular organelles or compartments for NP. By contrast, using a high-resolution ion beam system, discretized agglomerates of Au-NP were visualized inside the HeLa cell. This is consistent with the mechanism of entry of these NPs in the cellular space by endocytosis enclosed in endosomal vesicles. This approach shows RBS to be a powerful imaging technique assigning to nuclear microscopy unparalleled potential to assess nanoparticle distribution inside the cellular volume.

Steroid metabolism gene polymorphisms and their implications on breast and ovarian cancer prognosis.

A role for estrogen in the etiology of breast and ovarian cancers has been suggested; therefore, genetic polymorphisms in steroid metabolism genes could be involved in the carcinogenesis of these tumors. We have aimed to investigate the role of GSTP1 and CYP17 polymorphisms and their correlation with MSI (microsatellite instability) and LOH (loss of heterozygosity) in AR, ERß and CYP19 genes in women from Espírito Santo State, Brazil. The study population consisted of 107 female breast and 24 ovarian tumors. GSTP1 and CYP17 polymorphisms were detected by polymerase chain reaction (PCR) amplification followed by restriction fragment length polymorphism (RFLP) analysis while MSI and LOH were analyzed by PCR. GSTP1 and CYP17 polymorphisms alone were not associated with an increased risk for breast or ovarian tumors. However, when combined with MSI/LOH in AR, ERß and CYP19 genes, we were able to detect significant associations with the GSTP1 wild-type genotype in PR (progesterone receptor) negative breast cancers or the CYP17 wild-type genotype in ER (estrogen receptor) and PR-negative breast tumors. No associations with ovarian tumors were detected. Our results suggest that wild-type GSTP1 or CYP17 genes when combined with LOH/MSI in steroid metabolism genes may play a role in ER and/or PR negative breast cancers. These data support the hypothesis that genes related to steroid metabolism are important in the characterization of breast cancer and that the analysis of single polymorphisms may not be sufficient.

A multiproxy approach to evaluate biocidal treatments on biodeteriorated majolica glazed tiles.

The Fishing House located on the grounds of the Marquis of Pombal Palace, Oeiras, Portugal, was built in the 18th century. During this epoch, Portuguese gardens, such as the one surrounding the Fishing House, were commonly ornamented with glazed wall tile claddings. Currently, some of these outdoor tile panels are covered with dark colored biofilms, contributing to undesirable aesthetic changes and eventually inducing chemical and physical damage to the tile surfaces. Phylogenetic analyses revealed that the investigated biofilms are mainly composed of green algae, cyanobacteria and dematiaceous fungi. With the aim of mitigating biodeterioration, four different biocides (TiO2 nanoparticles, Biotin® T, Preventol® RI 80 and Albilex Biostat® ) were applied in situ to the glazed wall tiles. Their efficacy was monitored by visual examination, epifluorescence microscopy and DNA-based analysis. Significant changes in the microbial community composition were observed 4 months after treatment with Preventol® RI 80 and Biotin® T. Although the original community was inactivated after these treatments, an early stage of re-colonization was detected 6 months after the biocide application. TiO2 nanoparticles showed promising results due to their self-cleaning effect, causing the detachment of the biofilm from the tile surface, which remained clean 6 and even 24 months after biocide application. © 2013 Society for Applied Microbiology and John Wiley & Sons Ltd.

An integrated approach for assessing the bioreceptivity of glazed tiles to phototrophic microorganisms.

A laboratory-based methodology was designed to assess the bioreceptivity of glazed tiles. The experimental set-up consisted of multiple steps: manufacturing of pristine and artificially aged glazed tiles, enrichment of phototrophic microorganisms, inoculation of phototrophs on glazed tiles, incubation under optimal conditions and quantification of biomass. In addition, tile intrinsic properties were assessed to determine which material properties contributed to tile bioreceptivity. Biofilm growth and biomass were appraised by digital image analysis, colorimetry and chlorophyll a analysis. SEM, micro-Raman and micro-particle induced X-ray emission analyses were carried out to investigate the biodeteriorating potential of phototrophic microorganisms on the glazed tiles. This practical and multidisciplinary approach showed that the accelerated colonization conditions allowed different types of tile bioreceptivity to be distinguished and to be related to precise characteristics of the material. Aged tiles showed higher bioreceptivity than pristine tiles due to their higher capillarity and permeability. Moreover, biophysical deterioration caused by chasmoendolithic growth was observed on colonized tile surfaces.