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STATEMENT OF PROBLEM: Impregnated fibers require light polymerization; however, little information exists about how different protocols might affect the mechanical properties of reinforced denture base materials. PURPOSE: The purpose of this study was to compare the effects of pre- or postpolymerization of preimpregnated fibers on the flexural strength and elastic modulus of a reinforced autopolymerized and a heat-polymerized acrylic resin. MATERIAL AND METHODS: Seventy-two specimens were divided into 12 treatment groups (n=6), according to type of acrylic resin (autopolymerized or heat polymerized), type of reinforcement, and its pre- or postpolymerization. Impregnated glass fibers (Fibrex-Lab), unimpregnated glass fibers (Fibrante), and ribs made from a restorative composite resin (Z250) were used as reinforcements. The reinforcements were light polymerized either before or after incorporation and processing of the acrylic resins. Specimens were tested in 3-point load and the data were analyzed using 2-way ANOVA and Tukey post hoc test (alpha=.05). Specimens were further examined using light microscopy, atomic force microscopy, and scanning electron microscopy. RESULTS: Elastic modulus was significantly higher for heat-polymerized acrylic resins than for autopolymerized acrylic resins (P<.001). Prepolymerized fibers increased both flexural strength and elastic modulus of autopolymerized acrylic resins significantly more than postpolymerized fibers (P<.001); however, postpolymerized fibers yielded a higher elastic modulus than prepolymerized fibers for the heat-polymerized material (P<.001). CONCLUSIONS: Prepolymerized fibers improved the overall mechanical properties of reinforced autopolymerized acrylic resins more than postpolymerized fibers. However, postpolymerization of fibers yielded higher elastic modulus for reinforced heat-polymerized acrylics. [ABSTRACT FROM AUTHOR]

Abstract Background Herbaspirillum rubrisubalbicans was first identified as a bacterial plant pathogen, causing the mottled stripe disease in sugarcane. H. rubrisubalbicans can also associate with various plants of economic interest in a non pathogenic manner. Results A 21 kb DNA region of the H. rubrisubalbicans genome contains a cluster of 26 hrp/hrc genes encoding for the type three secretion system (T3SS) proteins. To investigate the contribution of T3SS to the plant-bacterial interaction process we generated mutant strains of H. rubrisubalbicans M1 carrying a Tn5 insertion in both the hrcN and hrpE genes. H. rubrisulbalbicans hrpE and hrcN mutant strains of the T3SS system failed to cause the mottled stripe disease in the sugarcane susceptible variety B-4362. These mutant strains also did not produce lesions on Vigna unguiculata leaves. Oryza sativa and Zea mays colonization experiments showed that mutations in hrpE and hrcN genes reduced the capacity of H. rubrisulbalbicans to colonize these plants, suggesting that hrpE and hrcN genes are involved in the endophytic colonization. Conclusions Our results indicate that the T3SS of H. rubrisubalbicans is necessary for the development of the mottled stripe disease and endophytic colonization of rice.

Abstract Background Metagenomics, the application of molecular genomics to consortia of non-cultivated microbes, has the potential to have a substantial impact on the search for novel industrial enzymes such as esterases (carboxyl ester hydrolases, EC 3.1.1.1) and lipases (triacylglycerol lipases, EC 3.1.1.3). In the current work, a novel lipase gene was identified from a fosmid metagenomic library constructed with the "prokaryotic-enriched" DNA from a fat-contaminated soil collected from a wastewater treatment plant. Results In preliminary screening on agar containing 1% tributyrin, 2661 of the approximately 500,000 clones in the metagenomic library showed activity. Of these, 127 showed activity on agar containing 1% tricaprylin, while 32 were shown to be true lipase producers through screening on agar containing 1% triolein. The clone with the largest halo was further characterized. Its lipase gene showed 72% identity to a putative lipase of Yersinia enterocolitica subsp. palearctica Y11. The lipase, named LipC12, belongs to family I.1 of bacterial lipases, has a chaperone-independent folding, does not possess disulfide bridges and is calcium ion dependent. It is stable from pH 6 to 11 and has activity from pH 4.5 to 10, with higher activities at alkaline pH values. LipC12 is stable up to 3.7 M NaCl and from 20 to 50°C, with maximum activity at 30°C over a 1 h incubation. The pure enzyme has specific activities of 1722 U/mg and 1767 U/mg against olive oil and pig fat, respectively. Moreover, it is highly stable in organic solvents at 15% and 30% (v/v). Conclusions The combination of the use of a fat-contaminated soil, enrichment of prokaryotic DNA and a three-step screening strategy led to a high number of lipase-producing clones in the metagenomic library. The most notable properties of the new lipase that was isolated and characterized were a high specific activity against long chain triacylglycerols, activity and stability over a wide range of pH values, good thermal stability and stability in water-miscible organic solvents and at high salt concentrations. These characteristics suggest that this lipase has potential to perform well in biocatalytic processes, such as for hydrolysis and synthesis reactions involving long-chain triacylglycerols and fatty acid esters.

Abstract Background CXCL12 is a chemokine that is constitutively expressed in many organs and tissues. CXCL12 promoter hypermethylation has been detected in primary breast tumours and contributes to their metastatic potential. It has been shown that the oestrogen receptor α (ESR1) gene can also be silenced by DNA methylation. In this study, we used methylation-specific PCR (MSP) to analyse the methylation status in two regions of the CXCL12 promoter and ESR1 in tumour cell lines and in primary breast tumour samples, and correlated our results with clinicopathological data. Methods First, we analysed CXCL12 expression in breast tumour cell lines by RT-PCR. We also used 5-aza-2'-deoxycytidine (5-aza-CdR) treatment and DNA bisulphite sequencing to study the promoter methylation for a specific region of CXCL12 in breast tumour cell lines. We evaluated CXCL12 and ESR1 methylation in primary tumour samples by methylation-specific PCR (MSP). Finally, promoter hypermethylation of these genes was analysed using Fisher's exact test and correlated with clinicopathological data using the Chi square test, Kaplan-Meier survival analysis and Cox regression analysis. Results CXCL12 promoter hypermethylation in the first region (island 2) and second region (island 4) was correlated with lack of expression of the gene in tumour cell lines. In the primary tumours, island 2 was hypermethylated in 14.5% of the samples and island 4 was hypermethylated in 54% of the samples. The ESR1 promoter was hypermethylated in 41% of breast tumour samples. In addition, the levels of ERα protein expression diminished with increased frequency of ESR1 methylation (p < 0.0001). This study also demonstrated that CXCL12 island 4 and ESR1 methylation occur simultaneously at a high frequency (p = 0.0220). Conclusions This is the first study showing a simultaneous involvement of epigenetic regulation for both CXCL12 and ESR1 genes in Brazilian women. The methylation status of both genes was significantly correlated with histologically advanced disease, the presence of metastases and death. Therefore, the methylation pattern of these genes could be used as a molecular marker for the prediction of breast cancer outcome.

Abstract Background ADAM33 protein is a member of the family of transmembrane glycoproteins composed of multidomains. ADAM family members have different activities, such as proteolysis and adhesion, making them good candidates to mediate the extracellular matrix remodelling and changes in cellular adhesion that characterise certain pathologies and cancer development. It was reported that one family member, ADAM23, is down-regulated by promoter hypermethylation. This seems to correlate with tumour progression and metastasis in breast cancer. In this study, we explored the involvement of ADAM33, another ADAM family member, in breast cancer. Methods First, we analysed ADAM33 expression in breast tumour cell lines by RT-PCR and western blotting. We also used 5-aza-2'-deoxycytidine (5azadCR) treatment and DNA bisulphite sequencing to study the promoter methylation of ADAM33 in breast tumour cell lines. We evaluated ADAM33 methylation in primary tumour samples by methylation specific PCR (MSP). Finally, ADAM33 promoter hypermethylation was correlated with clinicopathological data using the chi-square test and Fisher's exact test. Results The expression analysis of ADAM33 in breast tumour cell lines by RT-PCR revealed gene silencing in 65% of tumour cell lines. The corresponding lack of ADAM33 protein was confirmed by western blotting. We also used 5-aza-2'-deoxycytidine (5-aza-dCR) demethylation and bisulphite sequencing methodologies to confirm that gene silencing is due to ADAM33 promoter hypermethylation. Using MSP, we detected ADAM33 promoter hypermethylation in 40% of primary breast tumour samples. The correlation between methylation pattern and patient's clinicopathological data was not significantly associated with histological grade; tumour stage (TNM); tumour size; ER, PR or ERBB2 status; lymph node status; metastasis or recurrence. Methylation frequency in invasive lobular carcinoma (ILC) was 76.2% compared with 25.5% in invasive ductal carcinoma (IDC), and this difference was statistically significant (p = 0.0002). Conclusion ADAM33 gene silencing may be related to the discohesive histological appearance of ILCs. We suggest that ADAM33 promoter methylation may be a useful molecular marker for differentiating ILC and IDC.

Abstract Background Rice (Oryza sativa L.) germplasm represents an extraordinary source of genes that control traits of agronomic importance such as drought tolerance. This diversity is the basis for the development of new cultivars better adapted to water restriction conditions, in particular for upland rice, which is grown under rainfall. The analyses of subtractive cDNA libraries and differential protein expression of drought tolerant and susceptible genotypes can contribute to the understanding of the genetic control of water use efficiency in rice. Results Two subtractive libraries were constructed using cDNA of drought susceptible and tolerant genotypes submitted to stress against cDNA of well-watered plants. In silico analysis revealed 463 reads, which were grouped into 282 clusters. Several genes expressed exclusively in the tolerant or susceptible genotypes were identified. Additionally, proteome analysis of roots from stressed plants was performed and 22 proteins putatively associated to drought tolerance were identified by mass spectrometry. Conclusion Several genes and proteins involved in drought-response, as well as genes with no described homologs were identified. Genes exclusively expressed in the tolerant genotype were, in general, related to maintenance of turgor and cell integrity. In contrast, in the susceptible genotype, expression of genes involved in protection against cell damage was not detected. Several protein families identified in the proteomic analysis were not detected in the cDNA analysis. There is an indication that the mechanisms of susceptibility to drought in upland rice are similar to those of lowland varieties.

Objective: To estimate original wild-type BNT162b2 effectiveness against symptomatic Omicron infection among children 5-11 years of age., Methods: This prospective test-negative, case-control study was conducted in Toledo, southern Brazil, from June 2022 to July 2023. Patients were included if they were aged 5-11 years, sought care for acute respiratory symptoms in the public health system, and were tested for SARS-CoV-2 using reverse transcription polymerase chain reaction. In the primary analysis, we determined the effectiveness of two doses of original wild-type BNT162b2 against symptomatic COVID-19. The reference exposure group was the unvaccinated., Results: A total of 757 children were enrolled; of these, 461 (25 cases; 436 controls) were included in the primary analysis. Mean age was 7.4 years, 49.7 % were female, 34.6 % were obese, and 14.1 % had chronic pulmonary disease. Omicron accounted for 100 % of all identified SARS-CoV-2 variants with BA.5, BQ.1, and XBB.1 accounting for 35.7 %, 21.4 % and 21.4 %, respectively. The adjusted estimate of two-dose vaccine effectiveness against symptomatic Omicron was 3.1 % (95 % CI, -133.7 % to 61.8 %) after a median time between the second dose and the beginning of COVID-19 symptoms of 192.5 days (interquartile range, 99 to 242 days)., Conclusion: In this study with children 5-11 years of age, a two dose-schedule of original wild-type BNT162b2 was not associated with a significant protection against symptomatic Omicron infection after a median time between the second dose and the beginning of COVID-19 symptoms of 192 days, although the study may have been underpowered to detect a clinically important difference., Trial Registration Number: ClinicalTrials.gov number, NCT05403307 (https://classic., Clinicaltrials: gov/ct2/show/NCT05403307)., Competing Interests: Declaration of competing interest Rodrigues, Maltempi de Souza, Manfio, de Souza, Araujo, Cohen, Barbosa, Romeiro Silva, Sganzerla, Dias da Silva, Ferreira, Kunkel, Camargo, Sarturi, Guilhem, Oliveira, Lopes, Widmar, Barufi, Nunes da Silva, Gradia, Brandalize, Royer, Luiz, Baura, Abreu, and Poitevin report honoraria fee for working in this study from Hospital Moinhos de Vento. Rosa reports honoraria fee related to investigator activities from Pfizer, and research grants from Pfizer, MSD and Brazilian Ministry of Health. Falavigna reports honoraria fee related to investigator activities from Pfizer and MSD, consulting fees from Sanofi, Ultragenyx, Novartis, Alnylam, PTC and JCR, and honoraria for lectures from Janssen, Abbvie, Sanofi, Roche, Pfizer and Novartis.Valluri, Srivastava, Julião, Melone, Allen, Kyaw, Spinardi, Castillo, and McLaughlin are Pfizer empolyees. Kucharski, and Pedrotti have nothing to disclose., (Copyright © 2024. Published by Elsevier B.V.)

Background: To verify the influence of macrogeometry with healing chambers on the osseointegration of dental implants by analyzing implant stability quotient (ISQ) and evaluate the correlation between insertion torque and ISQ insertion with different macrogeometries., Methods: In total, 26 implants were installed in the posterior mandible of eight patients with sufficient bone height for the installation of implants measuring 3.5 mm in diameter and 9.0 mm in length. The implants were categorized according to two types of macrogeometry: a test group (GT) with 13 conical implants with healing chambers and a control group (GC) with 13 conical implants with conventional threads. To insert the implants, a bone drilling protocol was used up to a diameter of 3 mm with the last helical bur. The insertion torque of the implants was evaluated, followed by the measurement of ISQ at 0 (T-0), 7 (T-7), 14 (T-14), 21 (T-21), 28 (T-28), and 42 (T-42) days., Results: The mean insertion torque was 43 Ncm in both groups, without a significant difference. Moreover, no significant difference in the ISQ values was found between the groups at different time points (p > 0.05), except at T-7 (GT = 69.87±1.89 and GC = 66.48±4.49; p = 0.01). Although there was no significant difference, ISQ median values were higher in the GT group than GC group at 28 days (GT = 67.98 and GC = 63.46; p = 0.05) and 42 days (GT = 66.12 and GC = 60.33; p = 0.09). No correlation was found between the insertion torque and ISQ insertion (p > 0.05)., Conclusion: Furthermore, implants with a 3.5 mm diameter macrogeometry, with or without healing chambers, inserted with a drilling protocol up to 3 mm in diameter of the last helical bur, led to a similar secondary stability, with no difference in ISQ values. Although, implants with healing chamber demonstrates ascending values in the graph of ISQ, having a trend of faster osseointegration than implants without healing chambers. Both macrogeometries provide a similar primary stability to implants., Trial Registration: This study was registered retrospectively in ReBec (brazilian registry of clinical trials) under the number RBR-96n5×69, on the date of 19/06/2023., (© 2024. The Author(s).)

Background: Recent studies have revealed atypical features in the plastomes of the family Cactaceae, the largest lineage of succulent species adapted to arid and semi-arid regions. Most plastomes sequenced to date are from short-globose and cylindrical cacti, while little is known about plastomes of epiphytic cacti. Published cactus plastomes reveal reduction and complete loss of IRs, loss of genes, pseudogenization, and even degeneration of tRNA structures. Aiming to contribute with new insights into the plastid evolution of Cactaceae, particularly within the tribe Rhipsalideae, we de novo assembled and analyzed the plastomes of Lepismium cruciforme and Schlumbergera truncata, two South American epiphytic cacti., Methods and Results: Our data reveal many gene losses in both plastomes and the first loss of functionality of the trnT-GGU gene in Cactaceae. The trnT-GGU is a pseudogene in L. cruciforme plastome and appears to be degenerating in the tribe Rhipsalideae. Although the plastome structure is conserved among the species of the tribe Rhipsalideae, with tribe-specific rearrangements, we mapped around 200 simple sequence repeats and identified nine nucleotide polymorphism hotspots, useful to improve the phylogenetic resolutions of the Rhipsalideae. Furthermore, our analysis indicated high gene divergence and rapid evolution of RNA editing sites in plastid protein-coding genes in Cactaceae., Conclusions: Our findings show that some characteristics of the Rhipsalideae tribe are conserved, such as plastome structure with IRs containing only the ycf2 and two tRNA genes, structural degeneration of the trnT-GGU gene and ndh complex, and lastly, pseudogenization of rpl33 and rpl23 genes, both plastid translation-related genes., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

This study aimed to evaluate the impact of long-term liquid dairy manure (LDM) application on the activity and structure of soil bacterial and archaea communities in two cropping seasons over 1 year of a no-till crop rotation system. The experiment was run in a sandy clay loam texture Oxisol, in Brazil, including LDM doses of 60, 120, and 180 m 3  ha -1  year -1 , installed in 2005. Soil sampling was conducted during spring 2018 and autumn 2019 at 0-10-cm depth. Microbial biomass carbon and nitrogen, 16S rRNA gene sequencing, microbial respiration and quotient were performed. Over the 14-year period, LDM application increased soil microbial community activity. Analysis of 16S rRNA gene sequencing revealed dominance by Proteobacteria, Acidobacteria, and Actinobacteria phyla (67% in spring and 70% in autumn). Genera Pirulla and Nitrososphaera showed enrichment at LDM doses of 120 and 180 m 3  ha -1  year -1 doses, respectively. During spring, following black oat cropping, shifts in the relative abundance of Bacteroidetes, Proteobacteria, Firmicutes, Gemmatimonadetes, Verrucomicrobia, Chloroflexi, Actinobacteria, and AD3 phyla were observed due to LDM application, correlating with soil chemical indicators such as pH, K, Ca, Mn, and Zn. Our findings indicate that plant development strongly influences microbial community composition, potentially outweighing the impact of LDM. Our findings indicate that the application of liquid dairy manure alters the soil bacterial activity and community; however, this effect depends on the developing plant., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The RNA-Seq profiling of Herbaspirillum seropedicae SmR1 wild-type and ntrC mutant was performed under aerobic and three nitrogen conditions (ammonium limitation, ammonium shock, and nitrate shock) to identify the major metabolic pathways modulated by these nitrogen sources and those dependent on NtrC. Under ammonium limitation, H. seropedicae scavenges nitrogen compounds by activating transporter systems and metabolic pathways to utilize different nitrogen sources and by increasing proteolysis, along with genes involved in carbon storage, cell protection, and redox balance, while downregulating those involved in energy metabolism and protein synthesis. Growth on nitrate depends on the narKnirBDHsero&#95;2899nasA operon responding to nitrate and NtrC. Ammonium shock resulted in a higher number of genes differently expressed when compared to nitrate. Our results showed that NtrC activates a network of transcriptional regulators to prepare the cell for nitrogen starvation, and also synchronizes nitrogen metabolism with carbon and redox balance pathways., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Aquatic environments are subject to threats from multiple human activities, particularly through the release of untreated sanitary sewage into the coastal environments. These effluents contain a large group of natural or synthetic compounds referred to as emerging contaminants. Monitoring the types and quantities of toxic substances in the environment, especially complex mixtures, is an exhausting and challenging task. Integrative effect-based tools, such as biomarkers, are recommended for environmental quality monitoring programs. In this study, fish Poecilia vivipara were exposed for 24 and 96 h to raw untreated sewage diluted 33 % (v/v) in order to identify hepatic genes to be used as molecular biomarkers. Through a de novo hepatic transcriptome assembly, using Illumina MiSeq, 54,285 sequences were assembled creating a reference transcriptome for this guppy species. Transcripts involved in biotransformation systems, antioxidant defenses, ABC transporters, nuclear and xenobiotic receptors were identified and evaluated by qPCR. Sanitary sewage induced transcriptional changes in AhR, PXR, CYP2K1, CYP3A30, NQO1, UGT1A1, GSTa3, GSTmu, ST1C1, SOD, ABCC1 and SOX9 genes from liver of fish, particularly after 96 h of exposure. Changes in hepatic enzyme activities were also observed. The enzymes showed differences in fish exposed to both periods, while in the gills there was a prevalence of significant results after 96 h. The observed differences were associated to gender and/or to sewage exposure. The obtained results support the use of P. vivipara as sentinel and model organism for ecotoxicological studies and evidence the importance of understanding the differential responses associated to gender., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The objective is to present a clinical case of dental autotransplantation managed with surgery, orthodontics, endodontics, periodontics, and aesthetic rehabilitation. A 10-year-old boy sought treatment after avulsion of the maxillary left central incisor, which was not reimplanted. Based on anamnesis, clinical examination, and complementary examinations, agenesis of the maxillary and mandibular second premolars except the mandibular right second premolar was observed. After a multidisciplinary planning, the space in the maxillary left central incisor region was opened to receive the transplanted mandibular right second premolar. The receptor site was created in a single surgical procedure. Pulp necrosis was noted in the transplanted tooth, which was treated endodontically, and the agenesis spaces were closed using fixed orthodontic appliances. After removing the appliance, gingivectomy with osteotomy was performed in the maxillary right central incisor and the transplanted tooth regions to harmonize the height and shape of the gingival contour. Next, aesthetic readjustment was performed with tooth whitening, using office and home techniques, followed by microabrasion of the vestibular surface of the maxillary right central incisor. Direct composite resin restorations were placed in the maxillary incisors, and the teeth were rehabilitated using incisal and palatal guides. A multidisciplinary approach is essential for reestablishing the function and aesthetics of complex cases involving dental autotransplantation., Competing Interests: None declared., (The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/).)

Purpose: To analyze spatiotemporal trends in hospitalizations for cardiovascular diseases (CVD) sensitive to primary health care (PHC) among individuals aged 50-69 years in Paraná State, Brazil, from 2014 to 2019 and investigate correlations between PHC services and the Social Development Index., Methods: We conducted a cross-sectional ecological study using publicly available secondary data to analyze the municipal incidence of hospitalizations for CVD sensitive to PHC and to estimate the risk of hospitalization for this group of diseases and associated factors using hierarchical Bayesian spatiotemporal modeling with Markov chain Monte Carlo simulation., Results: There was a 5% decrease in the average rate of hospitalizations for PHC-sensitive CVD from 2014 to 2019. Regarding standardized hospitalization rate (SHR) according to population size, we found that no large municipality had an SHR >2. Likewise, a minority of these municipalities had SHR values of 1-2 (33%). However, many small and medium-sized municipalities had SHR values >2 (47% and 48%, respectively). A greater Social Development Index value served as a protective factor against hospitalizations, with a relative risk of 0.957 (95% credible interval, 0.929-0.984)., Conclusions: The annual risk of hospitalization decreased over time; however, small municipalities had the greatest rates of hospitalization, indicating an increase in health inequity. The inverse association between social development and hospitalizations for CVD sensitive to PHC raises questions about intersectionality in health care., (© 2024 Annals of Family Medicine, Inc.)

Statement of Problem: Little is known about the effect of different composite resin cements on the bond strength of prefabricated and milled glass fiber post-and-cores., Purpose: The purpose of this in vitro study was to compare the push-out bond strengths of root dentin and standard, relined, or computer-aided design and computer-aided manufacturing (CAD-CAM) glass fiber post-and-cores luted with conventional, universal, or self-adhesive composite resin cement., Material and Methods: Post spaces were prepared in 90 premolars. Standard and relined prefabricated and CAD-CAM-milled glass fiber post-and-cores were luted with conventional (RelyX ARC; 3M ESPE), universal (RelyX Ultimate; 3M ESPE), and self-adhesive (RelyX U200; 3M ESPE) composite resin cement (n=10). All specimens were thermally aged (5000 cycles at 5°C and 55°C with a 20-second dwell time) and mechanical fatigue (1 000 000 cycles at 2 Hz, 50-N load). The specimens were sectioned perpendicularly to the root long axis into 1-mm-thick sections (apical, middle, and cervical root thirds), the push-out bond strength was determined, and the mode of failure recorded. Data were submitted to the Kruskal-Wallis test followed by the Bonferroni-Dunn multiple comparison test (α=.05)., Results: Push-out bond strength values did not differ (P>.05) among posts at the same root thirds. Standard posts showed higher bond strength than CAD-CAM at the apical third for data grouped by composite resin cements (P<.001). The cervical root thirds had higher bond strength than the apical thirds (P<.05). The cervical, middle, and apical root thirds showed decreasing bond strength values for each type of post for data grouped by composite resin cement (P<.001). Adhesive failures at the dentin-cement interface were predominant., Conclusions: The type of posts and cements did not affect the bond strength of glass fiber posts. The cervical root thirds had better bonding performance than the apical thirds., (Copyright © 2024 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

One of the main factors limiting tilapia's production is the occurrence of infections caused by Aeromonas and Streptococcus species. This work intended to evaluate a bivalent vaccine against A. hydrophila and S. agalactiae by intraperitoneal (i.p) administration in Nile tilapia ( Oreochromis niloticus ) in Brazil. The study was carried out in two phases: one in the laboratory, on a small scale, and from the results obtained, the study was expanded to a large scale in a production system in cages. The vaccine proved to be safe and effective in laboratory tests, with a vaccine efficacy (VE) of 93.66%. However, in large-scale tests with 12,000 tilapias, the VE was 59.14%, with a better food conversion ratio (1.54 kg) in the vaccinated group compared to the control group (1.27 kg). These results corroborate the efficiency of this tested vaccine; however, they indicate the need for field tests to attest to real protection.

The appearance of new viruses and diseases has made the development of rapid and reliable diagnostic tests crucial. In light of it, we proposed a new method for assembling an electrochemical immunosensor, based on a one-step approach for selective layer formation. For this purpose, a mixture containing the immobilizing agent (polyxydroxybutyrate, PHB) and the recognition element (antibodies against SARS-CoV-2 nucleocapsid protein) was prepared and used to modify a screen-printed carbon electrode with electrodeposited graphene oxide, for the detection of SARS-CoV-2 nucleocapsid protein (N-protein). Under optimum conditions, N-protein was successfully detected in three different matrixes - saliva, serum, and nasal swab, with the lowest detectable values of 50 pg mL -1 , 1.0 ng mL -1, and 50 pg mL -1 , respectively. Selectivity was assessed against SARS-CoV-2 receptor-binding domain protein (RBD) and antibodies against yellow fever (YF), and no significant response was observed in presence of interferents, reinforcing the suitability of the proposed one-step approach for selective layer formation. The proposed biosensor was stable for up to 14 days, and the mixture was suitable for immunosensor preparation even after 60 days of preparation. The proposed assembly strategy reduces the cost, analysis time, and waste generation. This reduction is achieved through miniaturization, which results in the decreased use of reagents and sample volumes. Additionally, this approach enables healthcare diagnostics to be conducted in developing regions with limited resources. Therefore, the proposed one-step approach for selective layer formation is a suitable, simpler, and a reliable alternative for electrochemical immunosensing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Two novel copper(ii) complexes [Cu(2mni) 2 (H 2 O) 2 ](NO 3 ) 2 ·2H 2 O (1) and [Cu(2mni) 2 (NO 3 ) 2 ] (2), where 2mni is 2-methyl-5-nitroimidazole, were prepared and characterized in the solid state using single-crystal and powder X-ray diffraction analyses, EPR, electronic and vibrational spectroscopies (FTIR and Raman), and thermogravimetric methods. Both products present an elongated distorted octahedral geometry with axial Cu-O bond lengths of 2.606(14) and 2.593(15) Å, indicating semi-coordination. Density functional theory (DFT) calculations at the B3LYP/LANL2DZ theory level were used to study the electronic properties of 1 and 2. The Independent Gradient Model (IGM) was employed to determine the Intrinsic Bond Strength Index (IBSI) of the semi-coordination and to plot δ g isosurfaces for the electronic sharing between the metal center and ligands. A moderate to weak antibacterial activity against Escherichia coli cultures was found for 1 with a 50% growth inhibition (GI 50 ) value of 0.25 mmol L -1 . To the best of our knowledge, this is the first time that the semi-coordination analysis using IGM was carried out for a copper(ii) complex with axial elongation, finding a good correlation between the bond length and the IBSI, and the study was extended for a series of analogous complexes described in the literature., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (This journal is © The Royal Society of Chemistry.)

Trauma disproportionately affects vulnerable road users, especially the elderly. We analyzed the spatial distribution of elderly pedestrians struck by vehicles in the urban area of Maringa city, from 2014 to 2018. Hotspots were obtained by kernel density estimation and wavelet analysis. The relationship between spatial relative risks (RR) of elderly run-overs and the built environment was assessed through Qualitative Comparative Analysis (QCA). Incidents were more frequent in the central and southeast regions of the city, where the RR was up to 2.58 times higher. The QCA test found a significant association between elderly pedestrian victims and the presence of traffic lights, medical centers/hospitals, roundabouts and schools. There is an association between higher risk of elderly pedestrians collisions and specific elements of built environments in Maringa, providing fundamental data to help guide public policies to improve urban mobility aimed at protecting vulnerable road users and planning an age-friendly city.

Background: Evidence regarding effectiveness of BNT162b2 mRNA COVID-19 vaccine against Omicron in Latin America is limited. We estimated BNT162b2 effectiveness against symptomatic COVID-19 in Brazil when Omicron was predominant., Methods: This prospective test-negative, case-control study was conducted in Toledo, Brazil, following a mass COVID-19 vaccination with BNT162b2. Patients were included if they were aged ≥12 years, sought care for acute respiratory symptoms in the public health system between November 3, 2021 and June 20, 2022, and were tested for SARS-CoV-2 using RT-PCR. In the primary analysis, we determined the effectiveness of two doses of BNT162b2 against symptomatic COVID-19., Results: A total of 4,574 were enrolled; of these, 1,758 patients (586 cases and 1,172 controls) were included in the primary analysis. Mean age was 27.7 years, 53.8 % were women, and 90.1 % had a Charlson comorbidity index of zero. Omicron accounted for >97 % of all identified SARS-CoV-2 variants, with BA.1 and BA.2 accounting for 84.3 % and 12.6 %, respectively. Overall adjusted estimate of two-dose vaccine effectiveness against symptomatic COVID-19 was 46.7 % (95 %CI, 19.9 %-64.6 %) after a median time between the second dose and the beginning of COVID-19 symptoms of 94 days (IQR, 60-139 days). Effectiveness waned from 77.7 % at 7-29 days after receipt of a second dose to <30 % (non-significant) after ≥120 days., Conclusion: In a relatively young and healthy Brazilian population, two doses of BNT162b2 provided protection against symptomatic Omicron infection. However, this protection waned significantly over time, underscoring the need for boosting with variant-adapted vaccines in this population prior to waves of disease activity., Trial Registration Number: ClinicalTrials.gov number, NCT05052307 (https://clinicaltrials.gov/ct2/show/NCT05052307)., Competing Interests: Declaration of Competing Interest Rosa reports honoraria fee related to investigator activities from Pfizer, and research grants from Pfizer and Brazilian Ministry of Health. Falavigna reports honoraria fee related to investigator activities from Pfizer, consulting fees from Sanofi, Ultragenyx,Novartis, Alnylam, PTC and JCR, and honoraria for lectures from Janssen, Abbvie, Sanofi, Roche, Pfizer and Novartis. Manfio, Araujo, Cohen, Barbosa, Paula de Souza, Silva, Sganzerla, da Silva, Ferreira, Rodrigues, Maltempi de Souza de Souza, Oliveira, Gradia, Brandalize, Royer, and Luiz report honoraria fee for working in this study from Hospital Moinhos de Vento. Valluri, Srivastava, Julião, Melone, Allen, Kyaw, Spinardi, Castillo, and McLaughlin are Pfizer empolyees. Kucharski, and Pedrotti have nothing to disclose., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The COVID-19 pandemic resulted in the collapse of healthcare systems and led to the development and application of several approaches of wastewater-based epidemiology to monitor infected populations. The main objective of this study was to carry out a SARS-CoV-2 wastewater based surveillance in Curitiba, Southern Brazil Sewage samples were collected weekly for 20 months at the entrance of five treatment plants representing the entire city and quantified by qPCR using the N1 marker. The viral loads were correlated with epidemiological data. The correlation by sampling points showed that the relationship between the viral loads and the number of reported cases was best described by a cross-correlation function, indicating a lag between 7 and 14 days amidst the variables, whereas the data for the entire city presented a higher correlation (0.84) with the number of positive tests at lag 0 (sampling day). The results also suggest that the Omicron VOC resulted in higher titers than the Delta VOC. Overall, our results showed that the approach used was robust as an early warning system, even with the use of different epidemiological indicators or changes in the virus variants in circulation. Therefore, it can contribute to public decision-makers and health interventions, especially in vulnerable and low-income regions with limited clinical testing capacity. Looking toward the future, this approach will contribute to a new look at environmental sanitation and should even induce an increase in sewage coverage rates in emerging countries., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Cladophialophora exuberans is a filamentous fungus related to black yeasts in the order Chaetothyriales. These melanized fungi are known for their 'dual ecology', often occurring in toxic environments and also being frequently involved in human infection. Particularly Cladophialophora exuberans, C. immunda, C. psammophila, and Exophiala mesophila have been described with a pronounced ability to degrade aromatic compounds and xenobiotic volatiles, such as benzene, toluene, ethyl-benzene, and xylene, and are candidates for bioremediation applications. The objective of the present study is the sequencing, assembly, and description of the whole genome of C. exuberans focusing on genes and pathways related to carbon and toxin management, assessing the tolerance and bioremediation of lead and copper, and verifying the presence of genes for metal homeostasis. Genomic evaluations were carried out through a comparison with sibling species including clinical and environmental strains. Tolerance of metals was evaluated via a microdilution method establishing minimum inhibitory (MIC) and fungicidal concentrations (MFC), and agar diffusion assays. Heavy metal bioremediation was evaluated via graphite furnace atomic absorption spectroscopy (GFAAS). The final assembly of C. exuberans comprised 661 contigs, with genome size of 38.10 Mb, coverage of 89.9X and a GC content of 50.8%. In addition, inhibition of growth was shown at concentrations of 1250 ppm for copper and at 625 ppm for lead, using the MIC method. In the agar tests, the strain grew at 2500 ppm of copper and lead. In GFAAS tests, uptake capacities were observed of 89.2% and 95.7% for copper and lead, respectively, after 21 experimental days. This study enabled the annotation of genes involved in heavy metal homeostasis and also contributed to a better understanding of the mechanisms used in tolerance of and adaptation to extreme conditions., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interests., (Copyright © 2023 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Nitrogen is one of the most abundant elements in the biosphere, but its gaseous form is not biologically available to many organisms, including plants and animals. Diazotrophic microorganisms can convert atmospheric nitrogen into ammonia, a form that can be absorbed by plants in a process called biological nitrogen fixation (BNF). BNF is catalyzed by the enzyme nitrogenase, which not only reduces N 2 to NH 3 , but also reduces other substrates such as acetylene. The acetylene reduction assay (ARA) can be used to measure nitrogenase activity in diazotrophic organisms, either in symbiotic associations or in their free-living state. The technique uses gas chromatography to measure the reduction of acetylene to ethylene by nitrogenase in a simple, quick, and inexpensive manner. Here, we demonstrate how to: prepare nodulated soybean plants and culture free-living Azospirillum brasilense for the ARA, use the gas chromatograph to detect the ethylene formed, and calculate the nitrogenase activity based on the peaks generated by the chromatograph. The methods shown here using example organisms can be easily adapted to other nodulating plants and diazotrophic bacteria. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Acetylene reduction assay in root nodules Basic Protocol 2: Acetylene reduction assay using diazotrophic bacteria Basic Protocol 3: Calculation of nitrogenase activity Support Protocol 1: Production of acetylene from calcium carbide Support Protocol 2: Calibration of the gas chromatograph Support Protocol 3: Total protein quantification., (© 2023 Wiley Periodicals LLC.)

Encounters between flowers and invertebrates are key events for the functioning of tropical forests. Assessing the structure of networks composed of the interactions between those partners leads to a better understanding of ecosystem functioning and the effects of environmental factors on ecological processes. Gathering such data is, however, costly and time-consuming, especially in the highly diverse tropics. We aimed to provide a comprehensive repository of available flower-invertebrate interaction information for the Atlantic Forest, a South American tropical forest domain. Data were obtained from published works and "gray literature," such as theses and dissertations, as well as self-reports by co-authors. The data set has ~18,000 interaction records forming 482 networks, each containing between one and 1061 interaction links. Each network was sampled for about 200 h or less, with few exceptions. A total of 641 plant genera within 136 different families and 39 orders were reported, with the most abundant and rich families being Asteraceae, Fabaceae, and Rubiaceae. Invertebrates interacting with these plants were all arthropods from 10 orders, 129 families, and 581 genera, comprising 2419 morphotypes (including 988 named species). Hymenoptera was the most abundant and diverse order, with at least six times more records than the second-ranked order (Lepidoptera). The complete data set shows Hymenoptera interacting with all plant orders and also shows Diptera, Lepidoptera, Coleoptera, and Hemiptera to be important nodes. Among plants, Asterales and Fabales had the highest number of interactions. The best sampled environment was forest (~8000 records), followed by pastures and crops. Savanna, grasslands, and urban environments (among others) were also reported, indicating a wide range of approaches dedicated to collecting flower-invertebrate interaction data in the Atlantic Forest domain. Nevertheless, most reported data were from forest understory or lower strata, indicating a knowledge gap about flower-invertebrate interactions at the canopy. Also, access to remote regions remains a limitation, generating sampling bias across the geographical range of the Atlantic Forest. Future studies in these continuous and hard-to-access forested areas will yield important new information regarding the interactions between flowers and invertebrates in the Atlantic Forest. There are no copyright restrictions on the data set. Please cite this data paper if the data are used in publications and teaching events., (© 2022 The Ecological Society of America.)

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-positive pet owners are reported to be a risk factor for infection of their pets; however, the influence of the viral load and associated risks has not been fully established. This study aimed to assess potential association of viral load in owners with the presence of SARS-CoV-2 infection in their dogs. Of 20 SARS-CoV-2-positive pet owners from 13 families in Curitiba, Brazil, 5 of 22 (22.7%) dogs were positive for SARS-CoV-2. Viral presence was detected in oropharyngeal samples for 2 of 5 (40.0%) dogs at 8 and 9 days after the first positive sample. Detection of SARS-CoV-2 in these dogs was associated with higher viral loads in the owners and close owner contact. All 5 RT-qPCR-positive dogs had antibodies to at least one viral protein tested in the serological assay. Molecular detection of SARS-CoV-2 in dogs was statistically associated with clinical signs in owners such as cold, cough, or diarrhea (P = 0.039), number of positive persons in the household (P = 0.002), and higher viral load (P = 0.039). Such findings serve as a warning for risks of human to dog infection, mainly due to sharing beds and other close interactions without protection. In conclusion, people with coronavirus disease 2019 (COVID-19), particularly in households with multiple residents and high viral load, should take the same preventive measures when interacting with their dogs during self-isolation as they do with people., Competing Interests: Declaration of Competing Interest The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Brazil was the epicenter of worldwide pandemics at the peak of its second wave. The genomic/proteomic perspective of the COVID-19 pandemic in Brazil could provide insights to understand the global pandemics behavior. In this study, we track SARS-CoV-2 molecular information in Brazil using real-time bioinformatics and data science strategies to provide a comparative and evolutive panorama of the lineages in the country. SWeeP vectors represented the Brazilian and worldwide genomic/proteomic data from Global Initiative on Sharing Avian Influenza Data (GISAID) between February 2020 and August 2021. Clusters were analyzed and compared with PANGO lineages. Hierarchical clustering provided phylogenetic and evolutionary analyses of the lineages, and we tracked the P.1 (Gamma) variant origin. The genomic diversity based on Chao's estimation allowed us to compare richness and coverage among Brazilian states and other representative countries. We found that epidemics in Brazil occurred in two moments with different genetic profiles. The P.1 lineages emerged in the second wave, which was more aggressive. We could not trace the origin of P.1 from the variants present in Brazil. Instead, we found evidence pointing to its external source and a possible recombinant event that may relate P.1 to a B.1.1.28 variant subset. We discussed the potential application of the pipeline for emerging variants detection and the PANGO terminology stability over time. The diversity analysis showed that the low coverage and unbalanced sequencing among states in Brazil could have allowed the silent entry and dissemination of P.1 and other dangerous variants. This study may help to understand the development and consequences of variants of concern (VOC) entry., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Perico, De Pierri, Neto, Fernandes, Pedrosa, de Souza and Raittz.)

Chronic Chagasic cardiomyopathy (CCC), a progressive inflammatory and fibrosing disease, is the most prominent clinical form of Chagas disease, a neglected tropical disease caused by Trypanosoma cruzi infection. During CCC, the parasite remains inside the cardiac cells, leading to tissue damage, involving extensive inflammatory response and irregular fibrosis. Among the fibrogenic factors is transforming growth factor-β (TGF-β), a key cytokine controlling extracellular matrix synthesis and degradation. TGF-β is involved in CCC onset and progression, with increased serum levels and activation of its signaling pathways in the cardiac tissue, which crucially contributes to fibrosis. Inhibition of the TGF-β signaling pathway attenuates T. cruzi infection and prevents cardiac damage in an experimental model of acute Chagas disease. The aim of this study was to investigate the effect of TGF-β neutralization on T. cruzi infection in both in vitro and in vivo pre-clinical models, using the 1D11 monoclonal antibody. To this end, primary cultures of cardiac cells were infected with T. cruzi trypomastigote forms and treated with 1D11. For in vivo studies, 1D11 was administered in different schemes for acute and chronic phase models (Swiss mice infected with 10 4 parasites from the Y strain and C57BL/6 mice infected with 10 2 parasites from the Colombian strain, respectively). Here we show that the addition of 1D11 to cardiac cells greatly reduces cardiomyocyte invasion by T. cruzi and the number of parasites per infected cell. In both acute and chronic experimental models, T. cruzi infection altered the electrical conduction, decreasing the heart rate, increasing the PR interval and the P wave duration. The treatment with 1D11 reduced cardiac fibrosis and reversed electrical abnormalities improving cardiac performance. Taken together, these data further support the major role of the TGF-β signaling pathways in T. cruzi -infection and their biological consequences on parasite/host interactions. The therapeutic effects of the 1D11 antibody are promising and suggest a new possibility to treat cardiac fibrosis in the chronic phase of Chagas' heart disease by TGF-β neutralization., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer CC declared a shared affiliation with the authors RF, ES, WD, RA, and MW to the handling editor at the time of review., (Copyright © 2022 Ferreira, de Souza, Vilar-Pereira, Degrave, Abreu, Meuser-Batista, Ferreira, Ledbeter, Barker, Bailly, Feige, Lannes-Vieira, de Araújo-Jorge and Waghabi.)

In the present work, we report an innovative approach for immunosensors construction. The experimental strategy is based on the anchoring of biological material at screen-printed carbon electrode (SPE) modified with electrodeposited Graphene Quantum Dots (GQD) and polyhydroxybutyric acid (PHB). It was used as functional substract basis for the recognition site receptor-binding domain (RBD) from coronavirus spike protein (SARS-CoV-2), for the detection of Anti-S antibodies (AbS). SEM images and EDS spectra suggest an interaction of the protein with GQD-PHB sites at the electrode surface. Differential pulse voltametric (DPV) measurements were performed before and after incubation, in presence of the target, shown a decrease in voltametric signal of an electrochemical probe ([Fe(CN) 6 ] 3/4- ). Using the optimal experimental conditions, analytical curves were performed in PBS and human serum spiked with AbS showing a slight matrix effect and a relationship between voltametric signal and AbS concentration in the range of 100 ng mL -1 and 10 μg mL -1 . The selectivity of the proposed sensor was tested against yellow fever antibodies (YF) and the selective layer on the electrode surface did not interact with these unspecific antibodies. Eight samples of blood serum were analyzed and 87.5% of these total investigated provided adequate results. In addition, the present approach showed better results against traditional EDC/NHS reaction with enhancements in time and the possibility to develop an immunosensor in a single drop, since the proteins can be anchored prior to the electrode modification step., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Introduction: Real-world data on COVID-19 vaccine effectiveness are needed to validate evidence from randomized clinical trials. Accordingly, this study aims to evaluate, in a real-world setting in Brazil, the effectiveness of Pfizer-BioNTech BNT162b2 against symptomatic COVID-19 and COVID-19-related complications across diverse populations., Materials and Methods: A test-negative case-control study with follow-up of cases is currently being conducted in Toledo, a city in southern Brazil, following a mass COVID-19 vaccination campaign with BNT162b2. The study is being conducted among patients aged 12 years or older seeking care in the public health system with acute respiratory symptoms and tested for SARS-CoV-2 on reverse transcription polymerase chain reaction (RT-PCR). Cases are RT-PCR positive and controls RT-PCR negative. Test-positive cases are prospectively followed through structured telephone interviews performed at 15 days post-enrollment, and at 1, 3, 6, 9 and 12 months. Baseline demographic, clinical, and vaccination data are being collected by means of structured interviews and medical registry records reviews at the time of enrollment. All RT-PCR-positive samples are screened for mutations to identify SARS-CoV-2 variants., Ethics and Dissemination: The study protocol has been approved by the research ethics committee of all participant sites. Study findings will be disseminated through peer-reviewed publications and conference presentations., Trail Registration: Clinicatrials.gov: NCT05052307., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: Regis Goulart Rosa: has received honoraria related to investigator activities of the present study, research grants from the Brazilian Ministry of Health and Pfizer for COVID-19 studies, and honoraria for lectures from Novartis. Júlia Regazzini Spinardi: is Pfizer employee and owns Pfizer stocks. Kristen Allen: is Pfizer employee and owns Pfizer stocks. Josélia Manfio: is Moinhos de Vento employee Cintia Laura Pereira de Araujo: is Moinhos de Vento employee Mírian Cohen: is Moinhos de Vento employee and has received research grants from Medicago. Caroline Cabral Robinson: is Moinhos de Vento employee Daniel Sganzerla: has received honoraria related to statistical analysis activities of the present study. Diogo Rosas Ferreira: has received honoraria related to data managing activities of the present study. Emanuel Maltempi de Souza: has received supply of resources to perform SARS-COV-2 variant genotyping procedures of the present study. Jaqueline Carvalho de Oliveira: has received supply of resources to perform SARS-COV-2 variant genotyping procedures of the present study. Daniela Fiori Gradia: has received supply of resources to perform SARS-COV-2 variant genotyping procedures of the present study. Ana Paula Carneiro Brandalize: has received supply of resources to perform SARS-COV-2 variant genotyping procedures of the present study. Gabriela Almeida Kucharski: has no competing interests Fernando Pedrotti: has no competing interests Cristina de Oliveira Rodrigues: has received supply of resources to perform SARS-COV-2 variant genotyping procedures of the present study. Moe H. Kyaw: is Pfizer employee and owns Pfizer stocks. Graciela del Carmen Morales Castillo: is Pfizer employee and owns Pfizer stocks. Amit K. Srivastava: is Pfizer employee and owns Pfizer stocks. John M McLaughlin: is Pfizer employee and owns Pfizer stocks. Maicon Falavigna: has received honoraria related to investigator activities of the present study, consulting fees from Sanofi, Ultragenyx, Novartis, Alnylam, PTC and JCR, honoraria for lectures from Janssen, Abbvie, Sanofi, Roche, Pfizer and Novartis, and Payment for expert testimony from Merck Sharp & Dohme. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Dengue fever, caused by the dengue virus (DENV-1, -2, -3, and -4), affects millions of people in the tropical and subtropical regions worldwide. Severe dengue is correlated with high viraemia and cytokine storm, such as high levels of transforming growth factor-β1 (TGF-β1) in the patient's serum. Here, the TGF-β1 signaling was investigated in the context of in vitro viral clearance. Macrophages were infected with DENV-2 at MOI 5 and treated with the TGF-β receptor 1 and 2 inhibitor, GW788388. TGF-β1 expression, signal transduction and viral load were evaluated 48 h after DENV-2 infection by enzyme-linked immunoassay, immunofluorescence, and RT-qPCR assays. Total TGF-β1 level was reduced in 15% after DENV-2 infection, but the secretion of its biologically active form increased threefold during infection, which was followed by the phosphorylation of Smad2 protein. Phosphorylation of Smad2 was reduced by treatment with GW788388 and it was correlated with reduced cytokine production. Importantly, treatment led to a dose-dependent reduction in viral load, ranging from 6.6 × 10 5 RNA copies/ml in untreated cultures to 2.3 × 10 3 RNA copies/ml in cultures treated with 2 ng/ml of GW788388. The anti-TGF-β1 antibody treatment also induced a significant reduction in viral load to 1.6 × 10 3 RNA copies/ml. On the other hand, the addition of recombinant TGF-β1 in infected cultures promoted an increase in viral load to 7.0 × 10 6 RNA copies/ml. These results support that TGF-β1 plays a significant role in DENV-2 replication into macrophages and suggest that targeting TGF-β1 may represent an alternative therapeutic strategy to be explored in dengue infection., (© 2022 The Authors. Journal of Medical Virology published by Wiley Periodicals LLC.)

Gluconacetobacter diazotrophicus has been the focus of several studies aiming to understand the mechanisms behind this endophytic diazotrophic bacterium. The present study is the first global analysis of the early transcriptional response of exponentially growing G. diazotrophicus to iron, an essential cofactor for many enzymes involved in various metabolic pathways. RNA-seq, targeted gene mutagenesis and computational motif discovery tools were used to define the G. diazotrophicus fur regulon. The data analysis showed that genes encoding functions related to iron homeostasis were significantly upregulated in response to iron limitations. Certain genes involved in secondary metabolism were overexpressed under iron-limited conditions. In contrast, it was observed that the expression of genes involved in Fe-S cluster biosynthesis, flagellar biosynthesis and type IV secretion systems were downregulated in an iron-depleted culture medium. Our results support a model that controls transcription in G. diazotrophicus by fur function. The G. diazotrophicus fur protein was able to complement an E. coli fur mutant. These results provide new insights into the effects of iron on the metabolism of G. diazotrophicus , as well as demonstrate the essentiality of this micronutrient for the main characteristics of plant growth promotion by G. diazotrophicus .

Competing Interests: None disclosed.

Background and Aim: Acute kidney injury (AKI) is associated with a grave prognosis. A clinical assessment of kidney function can be performed based on the glomerular filtration rate (GFR). Cystatin C (CysC) can indicate the GFR or kidney function and its measurement is currently performed using immunological methods such as nephelometry, immunoturbidimetry, and enzyme-linked immunosorbent assays in human medicine. However, these techniques are not specific for use in veterinary medicine. This study aimed to validate an immunoturbidimetric assay for serum CysC (sCy) in dogs, determine the sCy reference intervals for healthy dogs, evaluate sCy stability in serum samples, and compare sCy with serum creatinine (sCr) in healthy dogs and dogs with AKI., Materials and Methods: Forty-three dogs were divided into a control group (n = 19) and an AKI group (n = 24). An immunoturbidimetric method including commercially available human CysC calibrated with canine CysC was used to evaluate canine serum samples., Results: An average recovery of 97% was observed for canine serum samples. The reference interval for CysC in healthy dogs was 0.57-1.29 mg/L. The sCy concentration in dogs with AKI was significantly higher (2.82 ± 1.46 mg/L) than in healthy dogs (0.93 ± 0.18 mg/L). Statistical analysis confirmed a strong correlation between sCy and sCr (r = 0.94; p < 0.05) in dogs with AKI., Conclusion: The immunoturbidimetric method of evaluating sCy yielded satisfactory results and can be used for canine samples when a species-specific calibrator is used. Furthermore, sCy is a reliable marker of renal dysfunction in dogs. It is best to store samples for sCy evaluation at temperatures between 4°C and 8°C., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Paes-Leme, et al.)

During the Zika fever outbreak in Brazil in 2015-2016, only some babies from infected mothers had teratogenic effects, suggesting that cofactors may influence congenital transmission. We investigated the ZIKV infection profile in explants and isolated cells from full-term human placenta to infection with the Brazilian Zika virus strain (ZIKV BR ) and the effect of coinfection with the Brazilian Human alphaherpesvirus 2 strain (HSV-2 BR ) on ZIKV replication. We found that the ZIKV BR infect the explants of amniotic and chorionic membranes, as well as chorionic villi core, but not the trophoblasts layer. It was also observed that ZIKV replication was higher in amniotic cells than chorionic and trophoblasts cells. Upon coinfection, the replication of ZIKV BR was reduced according to exposed HSV-2 BR load in trophoblasts cells and the levels of TNF-α and IL-6 cytokines were also reduced. These findings suggest that the placental cell types and HSV-2 BR coinfection may impact on ZIKV replication., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Transforming growth factor beta (TGF-β) is deeply involved on the pathogenesis of Chagas disease. Our group has been investigating the participation of this pleiotropic cytokine in different aspects of Chagas disease over the last 20 years. Important observations have been made, such as: (i) the ability of Trypanosoma cruzi in activating latent TGF-β; (ii) the potential involvement of TGF-β pathway on T. cruzi invasion of host cells; (iii) association of TGF-β with parasite intracellular replication; (iv) cardiac fibrosis development and maintenance; (v) disruption of Connexin-43 plaque structures and (vi) inflammation and immune response. In this perspective article we intend to discuss the advances of the potential use of new therapies targeting TGF-β to treat the cardiac alterations of Chagas disease-affected patients.

Background: Brazil has faced two simultaneous problems related to respiratory health: forest fires and the high mortality rate due to COVID-19 pandemics. The Amazon rain forest is one of the Brazilian biomes that suffers the most with fires caused by droughts and illegal deforestation. These fires can bring respiratory diseases associated with air pollution, and the State of Pará in Brazil is the most affected. COVID-19 pandemics associated with air pollution can potentially increase hospitalizations and deaths related to respiratory diseases. Here, we aimed to evaluate the association of fire occurrences with the COVID-19 mortality rates and general respiratory diseases hospitalizations in the State of Pará, Brazil., Methods: We employed machine learning technique for clustering k-means accompanied with the elbow method used to identify the ideal quantity of clusters for the k-means algorithm, clustering 10 groups of cities in the State of Pará where we selected the clusters with the highest and lowest fires occurrence from the 2015 to 2019. Next, an Auto-regressive Integrated Moving Average Exogenous (ARIMAX) model was proposed to study the serial correlation of respiratory diseases hospitalizations and their associations with fire occurrences. Regarding the COVID-19 analysis, we computed the mortality risk and its confidence level considering the quarterly incidence rate ratio in clusters with high and low exposure to fires., Findings: Using the k-means algorithm we identified two clusters with similar DHI (Development Human Index) and GDP (Gross Domestic Product) from a group of ten clusters that divided the State of Pará but with diverse behavior considering the hospitalizations and forest fires in the Amazon biome. From the auto-regressive and moving average model (ARIMAX), it was possible to show that besides the serial correlation, the fires occurrences contribute to the respiratory diseases increase, with an observed lag of six months after the fires for the case with high exposure to fires. A highlight that deserves attention concerns the relationship between fire occurrences and deaths. Historically, the risk of mortality by respiratory diseases is higher (about the double) in regions and periods with high exposure to fires than the ones with low exposure to fires. The same pattern remains in the period of the COVID-19 pandemic, where the risk of mortality for COVID-19 was 80% higher in the region and period with high exposure to fires. Regarding the SARS-COV-2 analysis, the risk of mortality related to COVID-19 is higher in the period with high exposure to fires than in the period with low exposure to fires. Another highlight concerns the relationship between fire occurrences and COVID-19 deaths. The results show that regions with high fire occurrences are associated with more cases of COVID deaths., Interpretation: The decision-make process is a critical problem mainly when it involves environmental and health control policies. Environmental policies are often more cost-effective as health measures than the use of public health services. This highlight the importance of data analyses to support the decision making and to identify population in need of better infrastructure due to historical environmental factors and the knowledge of associated health risk. The results suggest that The fires occurrences contribute to the increase of the respiratory diseases hospitalization. The mortality rate related to COVID-19 was higher for the period with high exposure to fires than the period with low exposure to fires. The regions with high fire occurrences is associated with more COVID-19 deaths, mainly in the months with high number of fires., Funding: No additional funding source was required for this study., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Authors.)

A promising strategy to overcome multidrug resistance is the use of inhibitors of ABC drug transporters. For this reason, we evaluated the polyoxovanadates (POVs) [V 10 O 28 ] 6- (V 10 ), [H 6 V 14 O 38 (PO 4 )] 5- (V 14 ), [V 15 O 36 Cl] 6- (V 15 ) and [V 18 O 42 I] 7- (V 18 ) as inhibitors of three major multidrug resistance-linked ABC transporters: P-glycoprotein (P-gp), ABCG2 and MRP1. All of the POVs selectively inhibited P-gp. V 10 and V 18 were the two most promising compounds, with IC 50 values of transport inhibition of 25.4 and 22.7 µm, respectively. Both compounds inhibited P-gp ATPase activity, with the same IC 50 value of 1.26 µm. V 10 and V 18 triggered different conformational changes in the P-gp protein with time-dependent inhibition, which was confirmed using the synthesized salt of V 10 with rhodamine B, RhoB-V 10 . The hydrophilic nature of POVs supports the hypothesis that these compounds target an unusual ligand-binding site, opening new possibilities in the development of potent modulators of ABC transporters., (© 2021 Federation of European Biochemical Societies.)

Competing Interests: None disclosed.

Crop diversity affects the processes of soil physical structuring and most likely provokes changes in the frequencies of soil microbial communities. The study was conducted for soil prokaryotic diversity sequencing 16S rDNA genes from a 25-year no-tillage experiment comprised of two crop systems: crop succession (Triticum aestivum-Glycine max) and rotation (Vicia sativa-Zea mays-Avena sativa-Glycine max-Triticum aestivum-Glycine max). The hypothesis was that a crop system with higher crop diversification (rotation) would affect the frequencies of prokaryotic taxa against a less diverse crop system (succession) altering the major soil functions guided by bacterial diversity. Soils in both crop systems were dominated by Proteobacteria (31%), Acidobacteria (23%), Actinobacteria (10%), and Gemmatimonadetes (7.2%), among other common copiotrophic soil bacteria. Crop systems did not affect the richness and diversity indexes of soil bacteria and soil archaea. However, the crop rotation system reduced only the frequencies of anaerobic metabolism bacteria Chloroacidobacteria, Holophagae, Spirochaetes, Euryarchaeota, and Crenarchaeota. It can be concluded that crop succession, a system that is poorer in root diversity over time, may have conditioned the soil to lower oxygen diffusion and built up ecological niches that suitable for anaerobic bacteria tolerating lower levels of oxygen. On the other hand, it appeared that crop rotation has restructured the soil over the years while enabling copiotrophic aerobic bacteria to dominate the soil ecosystem. The changes prompted by crop succession have implications for efficient soil organic matter decomposition, reduced greenhouse gas emissions, higher root activity, and overall soil productivity, which compromise to agriculture sustainability., (© 2021. Sociedade Brasileira de Microbiologia.)

We performed a large-scale severe acute respiratory syndrome coronavirus 2 screening campaign using 2 PCR-based approaches, coupled with variant genotyping, aiming to provide a safer environment for employees of Federal University in Curitiba, Brazil. We observed the rapid spread of the Gamma variant of concern, which replaced other variants in <3 months.

Purpose: To evaluate the color, surface roughness and microhardness of human dental enamel subjected to brushing with whitening dentifrices., Methods: The enamel samples (4 x 4 x 2 mm) were darkened using 0.2% chlorhexidine and black tea, and were randomly divided into seven groups (n= 15) for brushing cycles with only water; control; conventional dentifrice (Colgate Maximum Anticaries Protection); dentifrice containing 2% hydrogen peroxide (Colgate Luminous White Advanced); dentifrice containing sodium tripolyphosphate (Sensodyne True White); dentifrice containing activated charcoal (Black is White); and dentifrice containing blue covarine (Close Up White Now). The samples were submitted to 1,000 and 30,000 brushing cycles. The color values (CIE L*a*b*), surface roughness (Ra) and microhardness (Knoop) were recorded at four time points: baseline, after staining, after 1,000 and after 30,000 brushing cycles. The data were submitted to statistical analyses using paired t-test, ANOVA and Tukey., Results: The color analysis, roughness and microhardness results showed interaction between the effect of the dentifrices and the time point (P< 0.001). The bleaching dentifrices brought about color changes (ΔE) significantly greater than those obtained with conventional dentifrices and the control; however, there were no significant color changes among the bleaching dentifrices. When the color coordinates were evaluated individually, the L* values increased and the a* and b* values decreased throughout the experiment. The conventional toothpaste and the toothpastes containing blue covarine, activated charcoal or hydrogen peroxide promoted a statistically significant increase in the surface roughness of the samples after the last brushing cycle. The sodium tripolyphosphate dentifrice caused a progressive reduction in roughness. The microhardness increased statistically after 30,000 cycles for the conventional toothpaste, blue covarine and hydrogen peroxide. It was concluded that the whitening dentifrices lightened the samples, which evidenced greater luminosity and reduction in reddish and yellow tones. Moreover, they did not have deleterious effects on microhardness, and caused acceptable changes in surface roughness., Clinical Significance: Dentifrices with different whitening strategies changed the color of the samples, resulting in samples with greater luminosity and reduced reddish and yellowish tones. They also caused changes in surface roughness within the acceptable clinical limit and did not have a deleterious effect on the microhardness of dental enamel., Competing Interests: The authors declared no conflict of interest., (Copyright©American Journal of Dentistry.)

The genus Herbaspirillum is widely studied for its ability to associate with grasses and to perform biological nitrogen fixation. However, the bacteria of the Herbaspirillum genus have frequently been isolated from clinical samples. Understanding the genomic characteristics that allow these bacteria to switch environments and become able to colonize human hosts is essential for monitoring emerging pathogens and predicting outbreaks. In this work, we describe the sequencing, assembly, and annotation of the genome of H. frisingense AU14559 isolated from the sputum of patients with cystic fibrosis, and its comparison with the genomes of the uropathogenic strain VT-16-41 and the environmental strains GSF30, BH-1, IAC152, and SG826. The genes responsible for biological nitrogen fixation were absent from all strains except for GSF30. On the other hand, genes encoding virulence and host interaction factors were mostly shared with environmental strains. We also identified a large set of intrinsic antibiotic resistance genes that were shared across all strains. Unlike other strains, in addition to unique genomic islands, AU14559 has a mutation that renders the biosynthesis of rhamnose and its incorporation into the exopolysaccharide unfeasible. These data suggest that H. frisingense has characteristics that provide it with the metabolic diversity needed to infect and colonize human hosts.

Background: We report a genomic surveillance of SARS-CoV-2 lineages circulating in Paraná, southern Brazil, from March 2020 to April 2021. Our analysis, based on 333 genomes, revealed that the first variants detected in the state of Paraná in March 2020 were the B.1.1.33 and B.1.1.28 variants. The variants B.1.1.28 and B.1.1.33 were predominant throughout 2020 until the introduction of the variant P.2 in August 2020 and a variant of concern (VOC), Gamma (P.1), in January 2021. The VOC Gamma, a ramification of the B.1.1.28 lineage first detected in Manaus (northern Brazil), has grown rapidly since December 2020 and was thought to be responsible for the deadly second wave of COVID-19 throughout Brazil., Methods: The 333 genomic sequences of SARS-CoV-2 from March 2020 to April 2021 were generated as part of the genomic surveillance carried out by Fiocruz in Brazil Genomahcov Fiocruz. SARS-CoV-2 sequencing was performed using representative samples from all geographic areas of Paraná. Phylogenetic analyses were performed using the 333 genomes also included other SARS-CoV-2 genomes from the state of Paraná and other states in Brazil that were deposited in the GISAID. In addition, the time-scaled phylogenetic tree was constructed with up to 3 random sequences of the Gamma variant from each state in Brazil in each month of 2021. In this analysis we also added the sequences identified as the B.1.1.28 lineage of the Amazonas state and and the Gamma-like-II (P.1-like-II) lineage identified in different regions of Brazil., Results: Phylogenetic analyses of the SARS-CoV-2 genomes that were previously classified as the VOC Gamma lineage by WHO/PANGO showed that some genomes from February to April 2021 branched in a monophyletic clade and that these samples grouped together with genomes recently described with the lineage Gamma-like-II. Additionally, a new mutation (E661D) in the spike (S) protein has been identified in nearly 10% of the genomes classified as the VOC Gamma from Paraná in March and April 2021.Finally, we analyzed the correlation between the lineage and the Gamma variant frequency, age group (patients younger or older than 60 years old) and the clinical data of 86 cases from the state of Paraná., Conclusions: Our results provided a reliable picture of the evolution of the SARS-CoV-2 pandemic in the state of Paraná characterized by the dominance of the Gamma strain, as well as a high frequencies of the Gamma-like-II lineage and the S:E661D mutation. Epidemiological and genomic surveillance efforts should be continued to unveil the biological relevance of the novel mutations detected in the VOC Gamma in Paraná., (© 2021. The Author(s).)

Breast cancer (BC) is the most frequently diagnosed female cancer and second leading cause of death. Despite the discovery of many antineoplastic drugs for BC, the current therapy is not totally efficient. In this study, we investigated the potential of repurposing the well-known diabetes type II drug liraglutide to modulate epigenetic modifications in BC cells lines in vitro and in vivo via Ehrlich mice tumors models. The in vitro results revealed a significant reduction on cell viability, migration, DNMT activity and displayed lower levels of global DNA methylation in BC cell lines after liraglutide treatment. The interaction between liraglutide and the DNMT enzymes resulted in a decrease profile of DNA methylation for the CDH1, ESR1 and ADAM33 gene promoter regions and, consequently, increased their gene and protein expression levels. To elucidate the possible interaction between liraglutide and the DNMT1 protein, we performed an in silico study that indicates liraglutide binding in the catalytic cleft via hydrogen bonds and salt bridges with the interdomain contacts and disturbs the overall enzyme conformation. The in vivo study was also able to reveal that liraglutide and the combined treatment of liraglutide and paclitaxel or methotrexate were effective in reducing tumor growth. Moreover, the modulation of CDH1 and ADAM33 mouse gene expression by DNA demethylation suggests a role for liraglutide in DNMT activity in vivo. Altogether, these results indicate that liraglutide may be further analysed as a new adjuvant treatment for BC., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Herbaspirillum seropedicae is a β-proteobacterium that establishes as an endophyte in various plants. These bacteria can consume diverse carbon sources, including hexoses and pentoses like D-xylose. D-xylose catabolic pathways have been described in some microorganisms, but databases of genes involved in these routes are limited. This is of special interest in biotechnology, considering that D-xylose is the second most abundant sugar in nature and some microorganisms, including H. seropedicae, are able to accumulate poly-3-hydroxybutyrate when consuming this pentose as a carbon source. In this work, we present a study of D-xylose catabolic pathways in H. seropedicae strain Z69 using RNA-seq analysis and subsequent analysis of phenotypes determined in targeted mutants in corresponding identified genes. G5B88&#95;22805 gene, designated xylB, encodes a NAD + -dependent D-xylose dehydrogenase. Mutant Z69∆xylB was still able to grow on D-xylose, although at a reduced rate. This appears to be due to the expression of an L-arabinose dehydrogenase, encoded by the araB gene (G5B88&#95;05250), that can use D-xylose as a substrate. According to our results, H. seropedicae Z69 uses non-phosphorylative pathways to catabolize D-xylose. The lower portion of metabolism involves co-expression of two routes: the Weimberg pathway that produces α-ketoglutarate and a novel pathway recently described that synthesizes pyruvate and glycolate. This novel pathway appears to contribute to D-xylose metabolism, since a mutant in the last step, Z69∆mhpD, was able to grow on this pentose only after an extended lag phase (40-50 h). KEY POINTS: • xylB gene (G5B88&#95;22805) encodes a NAD + -dependent D-xylose dehydrogenase. • araB gene (G5B88&#95;05250) encodes a L-arabinose dehydrogenase able to recognize D-xylose. • A novel route involving mhpD gene is preferred for D-xylose catabolism., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)