Microscopic Characterization of the Infectious Process, ROS Production, and Fungi Cellular Death of Alternaria alternata on Tangerine Resistant to QoIs.

Quinone outside inhibitor (QoI) fungicide resistance in Alternaria alternata populations was reported in Brazil for the first time in 2019, in São Paulo orchards, and the mutation G143A in cytochrome b (cytb) was found in resistant isolates. Our study investigated the infectious process, production of reactive oxygen species (ROS), and fungal cell death in resistant (QoI-R) and sensitive (QoI-S) A. alternata pathotype tangerine (Aapt) isolates. Morphological characterization of Aapt isolates was performed using confocal laser scanning microscopy (CLSM). Alternaria brown spot (ABS) symptoms were produced by Aapt isolates on tangelo cv. BRS Piemonte. Germination of QoI-R conidia and production of germ tubes on tangelo leaflets treated with 100 µg mL-1 of pyraclostrobin 18 h after inoculation (hai) was observed using scanning electron microscopy (SEM). At the same time, QoI-S conidial germination was inhibited on tangelo leaflets treated with pyraclostrobin. ROS production and cell death in Aapt isolates at high fungicide concentrations were observed using CLSM. QoI-S conidia exhibited high ROS production, indicating high oxidative stress. When dyed with propidium iodate (PI), QoI-S conidia emitted red fluorescence, showing cell death and confirming their sensitive phenotype. In contrast, QoI-R conidia neither produced ROS nor exhibited red fluorescence, indicating no cell death and confirming their resistant phenotype. Therefore, our findings evidence that microscopic techniques may help characterize events during fungi-plant interactions, ROS production, cell death, and Aapt phenotypes resistant and sensitive to QoIs using fluorometric protocols.

A global overview of enamel microabrasion for white spot lesions: a bibliometric review.

This study aimed to identify and analyze articles on enamel microabrasion for the treatment of white spot lesions. A search was conducted on the Web of Science. The following parameters were recorded and analyzed: number of citations, year, journal, impact factor, study design, theme, country and continent, institution, authors, and keywords. Data was analyzed using VOSviewer software. The initial search resulted in 1,126 documents, of which 94 articles were included. The highest number of citations an article received was 65. The oldest article was published in 1975, and the most recent in 2023. The most frequent study design was case report (n = 42). Regarding the themes, it was observed that the main objective of the studies was to evaluate the clinical performance of enamel microabrasion (n = 75), primarily using Opalustre (Ultradent Products Inc., South Jordan, UT, USA) (n = 37) for treating white stains caused by dental fluorosis (n = 41). Most articles originated from Latin America (n = 31), mainly from Brazil (n = 26). The most frequent author was Sundfeld RH (n = 10). This study reveals research trends in the field of enamel microabrasion. The publications were mainly case reports/series using Opalustre for the removal of fluorosis stains.

Identification and serological responses to a novel Plasmodium vivax merozoite surface protein 1 (PvMSP-1) derived synthetic peptide: a putative biomarker for malaria exposure.

Background: The integration of diagnostic methods holds promise for advancing the surveillance of malaria transmission in both endemic and non-endemic regions. Serological assays emerge as valuable tools to identify and delimit malaria transmission, serving as a complementary method to rapid diagnostic tests (RDT) and thick smear microscopy. Here, we evaluate the potential of antibodies directed against peptides encompassing the entire amino acid sequence of the PvMSP-1 Sal-I strain as viable serological biomarkers for P. vivax exposure. Methods: We screened peptides encompassing the complete amino acid sequence of the Plasmodium vivax Merozoite Surface Protein 1 (PvMSP-1) Sal-I strain as potential biomarkers for P. vivax exposure. Here, immunodominant peptides specifically recognized by antibodies from individuals infected with P. vivax were identified using the SPOT-synthesis technique followed by immunoblotting. Two 15-mer peptides were selected based on their higher and specific reactivity in immunoblotting assays. Subsequently, peptides p70 and p314 were synthesized in soluble form using SPPS (Solid Phase Peptide Synthesis) and tested by ELISA (IgG, and subclasses). Results: This study unveils the presence of IgG antibodies against the peptide p314 in most P. vivax-infected individuals from the Brazilian Amazon region. In silico B-cell epitope prediction further supports the utilization of p314 as a potential biomarker for evaluating malaria transmission, strengthened by its amino acid sequence being part of a conserved block of PvMSP-1. Indeed, compared to patients infected with P. falciparum and uninfected individuals never exposed to malaria, P. vivax-infected patients have a notably higher recognition of p314 by IgG1 and IgG3.

First report of Colletotrichum plurivorum and Colletotrichum truncatum causing anthracnose on melon plants in Brazil.

Melon (Cucumis melo L.) is an economically important crop in Brazil, with an annual production of 699.281 tons (FAO 2024). Fungal diseases are one of the biggest problems in melon production, and melon growers in northeastern Brazil have reported over 80% of plants showing anthracnose symptoms in the fields during rainy seasons. Plants were wilted, displaying brown necrotic lesions and water-soaked spots with yellowish edges on the leaves and vines. Melon fruits displayed necrotic lesions on the outside. From June 2022 to June 2023, melon leaves (varieties Yellow, Galia, and Cantaloupe) from anthracnose-symptomatic plants were collected in four melon farms located in the municipalities of Afonso Bezerra, Mossoró, Tibau, and Upanema in the state of Rio Grande do Norte. Small fragments of symptomatic leaves were disinfected in 70% ethanol (30 sec) and 2.5 % sodium hypochlorite (1 min), rinsed in sterile distilled water, and plated on PDA Petri dishes with tetracycline (0.05g/liter). Plates were maintained in a bio-oxygen demand incubator (BOD) for 3 days at 28 ± 2 °C, under a 12 hr photoperiod. Eleven representative fungal colonies resembling Colletotrichum spp. were selected and monosporically grown on PDA for seven days for morphology, pathogenicity, and molecular analyses.ight colonies showed pinkish-dark brown with acervuli in the center and cottony mycelium, and showing black edges in some isolates, resembling C. plurivorum (Zhang et al. 2023). Conidia from those colonies were hyaline, cylindrical with obtuse ends, and 17.76 x 7.06 µm, n= 50. Three colonies developed pinkish-gray mycelia with numerous black microsclerotia, and the conidia were hyaline, falcate, and 27.38 x 4.10 µm, n= 50, resembling C. truncatum (Yu et al. 2023). The total DNA of the eleven isolates was extracted, and the internal transcribed space (ITS), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), actin (ACT), ß-tubulin (TUB), and chitin synthase 1 (CHS-1) regions were partially amplified by PCR. Amplicons were sequenced and deposited to Genbank (Table eXtra1). A phylogenetic tree was built with the Maximum likelihood method with the concatenated sequences of the five partial gene sequences on Software MEGA (Version 11.0.10) (Tamura et al. 2021). The isolates CML5, CML8, CML9, CML10, CML11, CML14, CML15, and CML25 were grouped with Colletotrichum plurivorum CBS 125474 (orchidearum complex), and the isolates CML26, CML27 and CML28 with Colletotrichum truncatum CBS 15:35 (truncatum complex) with 87 % e 97 % of Bootstrap support, respectively. C. plurivorum was detected in four farms visited (we selected two representative isolates per farm), while C. truncatum isolates were all from the farm in Afonso Bezerra municipality. A pathogenicity test was performed following the method of Baishuan et al. (2023), micro-injuries were made in leaves of melon seedlings 'Goldex Yellow' and inoculated with a spore suspension of colonies with seven days of growth (106 spore/mL) of each isolate and sprayed to the point of dripping. Sterile water was used as mock. After nine days, anthracnose symptoms similar to those observed in the field were seen in all inoculated leaves, while no symptom was observed in the leaves of the mock plants. The pathogens were reisolated and their identification was confirmed by morphology and sequencing. Five seedlings were inoculated per isolate and mock, the assay was repeated, and the same results were observed. The species C. plurivorum has already been reported to cause disease in Cucumbers in Brazil (Silva et al. 2023) and C. plurivorum and C. truncatum in Citrullus lanatus in China (Guo et al. 2022). To the best of our knowledge, this is the first report of C. plurivorum and C. truncatum causing anthracnose in melon plants in Brazil.

Isolation and Characterization of Two Novel Genera of Jumbo Bacteriophages Infecting Xanthomonas vesicatoria Isolated from Agricultural Regions in Mexico.

Bacterial spot is a serious disease caused by several species of Xanthomonas affecting pepper and tomato production worldwide. Since the strategies employed for disease management have been inefficient and pose a threat for environmental and human health, the development of alternative methods is gaining relevance. The aim of this study is to isolate and characterize lytic phages against Xanthomonas pathogens. Here, we isolate two jumbo phages, named XaC1 and XbC2, from water obtained from agricultural irrigation channels by the enrichment technique using X. vesicatoria as a host. We determined that both phages were specific for inducing the lysis of X. vesicatoria strains, but not of other xanthomonads. The XaC1 and XbC2 phages showed a myovirus morphology and were classified as jumbo phages due to their genomes being larger than 200 kb. Phylogenetic and comparative analysis suggests that XaC1 and XbC2 represent both different and novel genera of phages, where XaC1 possesses a low similarity to other phage genomes reported before. Finally, XaC1 and XbC2 exhibited thermal stability up to 45 °C and pH stability from 5 to 9. All these results indicate that the isolated phages are promising candidates for the development of formulations against bacterial spot, although further characterization is required.

Nitrogen and Silicon Contribute to Wheat Defense's to Pyrenophora tritici-repentis, but in an Independent Manner.

Nitrogen (N) and silicon (Si) are mineral elements that have shown a reduction in the damage caused by tan spot (Pyrenophora tritici-repentis (Ptr)) in wheat. However, the effects of these elements were studied separately, and the N and Si interaction effect on wheat resistance to tan spot remains elusive. Histocytological and biochemical defense responses against Ptr in wheat leaves treated with Si (+Si) at low (LN) and high N (HN) inputs were investigated. Soil amendment with Si reduced the tan spot severity in 18% due to the increase in the leaf Si concentration (around 30%), but it was affected by the N level used. The superoxide dismutase (SOD) activity was higher in +Si plants and inoculated with Ptr, leading to early and higher H2O2 and callose accumulation in wheat leaf. Interestedly, phenylalanine ammonia-lyase (PAL) activity was induced by the Si supplying, being negatively affected by the HN rate. Meanwhile, catalase (CAT), and peroxidase (POX) activities showed differential response patterns according to the Si and N rates used. Tan spot severity was reduced by both elements, but their interaction does not evidence synergic effects in this disease's control. Wheat plants from -Si and HN and +Si and LN treatments recorded lower tan spot severity.

Expanding on expansus: a new species of Scaphanocephalus from North America and the Caribbean based on molecular and morphological data.

Members of the genus Scaphanocephalus mature in accipitrids, particularly osprey, Pandion haliaetus, with metacercaria causing Black Spot Syndrome in reef fishes. In most of the world, only the type species, Scaphanocephalus expansus (Creplin, 1842) has been reported. Recent molecular studies in the Western Atlantic, Mediterranean and Persian Gulf reveal multiple species of Scaphanocephalus, but have relied on 28S rDNA, mainly from metacercariae, which limits both morphological identification and resolution of closely related species. Here we combine nuclear rDNA with mitochondrial sequences from adult worms collected in osprey across North America and the Caribbean to describe species and elucidate life cycles in Scaphanocephalus. A new species described herein can be distinguished from S. expansus based on overall body shape and size. Phylogenetic analysis of the whole mitochondrial genome of Scaphanocephalus indicates a close relationship with Cryptocotyle. We conclude that at least 3 species of Scaphanocephalus are present in the Americas and 2 others are in the Old World. Specimens in the Americas have similar or identical 28S to those in the Mediterranean and Persian Gulf, but amphi-Atlantic species are unlikely in light of divergence in cytochrome c oxidase I and the lack of amphi-Atlantic avian and fish hosts. Our results provide insight into the geographic distribution and taxonomy of a little-studied trematode recently linked to an emerging pathology in ecologically important reef fishes.

Infiltração resinosa em lesões brancas cariosas e não cariosas: relato de casos / Infiltración resinosa em lesiones blancas cariosas y no cariosas: Reporte de Casos / Resinous infiltration in carious and non-carious white lesions: Case Reports

Introdução: O processo de desmineralização proveniente da cárie leva à formação de manchas brancas que são a primeira manifestação visível da doença. Os infiltrantes resinosos surgem como uma alternativa para o tratamento dessas lesões não cavitadas, pois sãoresinas de baixa viscosidade e faz parte dos procedimentos que visam uma odontologia menos invasiva. Objetivo: Este estudo objetiva relatar a experiência clínica no uso de infiltrante resinoso, Icon, em lesões de mancha branca de etiologias cariosa e não cariosa em dois pacientes distintos, insatisfeitos com a estética do seu sorriso. Descrição do caso: Ao exame clínico foi observado nos pacientes com lesões brancas. No primeiro paciente, de 14 anos, verificou-se a presença da atividade de cárie e micro cavitações. Dessa forma, o plano de tratamento perpassou orientação de higiene oral supervisionada, aplicação de verniz fluoretado, Enamelast, semanal, adequação do meio com restaurações em resina, e só então o uso do infiltrante. A segunda paciente, de 11 anos, já possuía saúde bucal adequada e tinha queixa estética devido à lesão branca não cariosa, hipoplasia, cujo plano de tratamento foi à utilização do Icon apenas. Conclusões: Nos dois casos obteve-se melhoria estética considerável com este procedimento microinvasivo. Melhoria na saúde bucal, aliado à devolução da estética, pode ser observada com o uso de infiltrantes resinosos (AU).

Introduction: The process of demineralization resulting from caries leads to the formation of white spots that are the first visible manifestation of the disease. Resin infiltrants appear as an alternative for the treatment of these non-cavitated lesions, since they are low viscosity resins and are part of the procedures that aim at a less invasive dentistry.Objective:This study aims to report the clinical experience in the use of a resin infiltrant, Icon,in white spot lesions of carious and non-carious etiologies in two different patients who were dissatisfied with the esthetics of their smiles.Methodology: On clinical examination, white lesions were observed in both patients. In the first patient,14 years old,the presence of caries activity and micro cavitations was verified. Thus, the treatment plan included supervised oral hygiene guidance, weekly application of fluoride varnish,Enamelast, adaptation of the environment with resin restorations, and, onlythen, the use of the infiltrant, Icon. The second patient, 11 years old,already had adequate oral health and had an esthetic complaint due to a non-carious white lesion, hypoplasia, whose treatment plan consisted of the use of Icon only.Conclusions:In both cases considerable esthetic improvement was obtained with this microinvasive procedure. Improvement in oral health, combined with the return of esthetics, can be observed with the use of resin infiltrants (AU).

Introducción: El proceso de desmineralización resultante de la caries conduce a la formación de manchas blancas, que son la primera manifestación visible de la enfermedad. Losinfiltrantes de resina aparecen como una alternativa para el tratamiento de estas lesiones no cavitadas, pues se tratan de resinas de baja viscosidad y forman parte de los procedimientos que buscan una odontología menos invasiva. Objetivo:Este estudio tiene como objetivo relatar la experiencia clínica en el uso del infiltrante de resina,Iconen lesiones de mancha blanca de etiologías cariosas y no cariosas en dos pacientes diferentes, insatisfechos con la estética de sus sonrisas.Metodología: En el examen clínico se observaron lesiones blancas en ambos pacientes. En el primer paciente, 14 añosse verificó la presencia de actividad de caries y micro cavitaciones. De ese modo, el plan de tratamiento incluyó la orientación supervisada de la higiene bucal,la aplicación semanal de barniz de flúor (Enamelast), la adaptación del entorno con restauraciones de resina y, sólo después, el uso del infiltrante,Icon. La segunda paciente, 11 años,ya tenía una salud bucal adecuada y presentaba una queja estética debido a una lesión blanca no cariosa, hipoplasia, cuyo plan de tratamiento fue el uso exclusivo de Icon. Conclusiones: En ambos casos, se consiguió una mejora estética considerable con este procedimiento microinvasivo. La mejora de la salud bucal, unida a la recuperación de la estética, puede observarse con el uso de infiltrantes de resina (AU).

Prevalence and associated risk factors of hepatitis C antibody and RNA among people who inject drugs in Puerto Rico.

INTRODUCTION: As a public health crisis, hepatitis C viral infection (HCV) is highly prevalent among people who inject drugs (PWID). We aimed to assess factors associated with HCV antibody (Ab) and HCV ribonucleic acid (RNA) positivity among PWID in Puerto Rico. METHODS: The study recruited a total of 150 persons in rural and peri-urban community settings through the respondent-driven sampling method and administered a structured questionnaire. We conducted HIV and HCV testing using dried blood spots (DBS). We examined correlates of HCV infection with sociodemographics, drug use patterns, and injection practices using regression in bivariate and multivariable analysis. RESULTS: Of the 150 participants, 89 % were male; 11 % were female; 72 % identified as mixed race; and the median duration of drug injection was 17.8 years. The mean age was 43.1 years, with 64 % of the population being from 23 to 45 years old. Among study participants (n = 150), the prevalence of HCV Ab was 73 %, and HCV RNA was 53 %. Factors significantly associated with HCV Ab and RNA included older age, increasing years of injection, incarceration, injecting other people, and identifying as Black. The belief that syringe air blowing reduces HCV transmission was also independently associated with HCV Ab positivity. CONCLUSIONS: Our findings regarding risk factors associated with HCV infection show the need to enhance prevention and control strategies for reducing transmission among PWID. Direct-acting antiviral treatment, sustained access to harm reduction, and culturally tailored services will be required to substantially reduce rates of HCV. Community-based treatment models and treatment in correctional settings are needed.

Draft genome sequence data of Nothopassalora personata, peanut foliar pathogen from Argentina.

Late leaf spot (LLS) caused by the Ascomycete Nothopassalora personata (N.p.) (Syn. Cercosporidium personatum) is the main foliar disease of peanuts in Argentina and in peanut producing areas of the world, causing up to 70% yield losses. The extremely slow growth of this fungus in culture, that takes around one month to form a 1 cm colony (0.45 mm/day), and the lack of adequate young tissues from where to extract nucleic acids, have hindered genetic studies of this pathogen. Here, we report the first genome sequence of a N. personata isolate from South America, as well as genetic variants on its conserved genes, and the complete sequence of its mating-type locus MAT1-2 idiomorph. The N. personata isolate IPAVE 0302 was obtained from peanut leaves in Córdoba, Argentina. The whole genome sequencing of IPAVE 0302 was performed as paired end 150 bp NovaSeq 6000 and de novo assembled. Clean reads were mapped to the reference genome for this species NRRL 64463 and the genetic variants on highly conserved genes and throughout the genome were analyzed. Sequencing data were submitted to NCBI GenBank Bioproject PRJNA948451, accession number SRR23957761. Additional Fasta files are available from Harvard Dataverse (https://doi.org/10.7910/DVN/9AGPMG and https://doi.org/10.7910/DVN/YDO3V6). The data reported here will be the basis for the analysis of genetic diversity of the LLS pathogen of peanut in Argentina, information that is critical to make decisions on management strategies.

TRPA1 channel mediates methylglyoxal-induced mouse bladder dysfunction.

Introduction: The transient receptor potential ankyrin 1 channel (TRPA1) is expressed in urothelial cells and bladder nerve endings. Hyperglycemia in diabetic individuals induces accumulation of the highly reactive dicarbonyl compound methylglyoxal (MGO), which modulates TRPA1 activity. Long-term oral intake of MGO causes mouse bladder dysfunction. We hypothesized that TRPA1 takes part in the machinery that leads to MGO-induced bladder dysfunction. Therefore, we evaluated TRPA1 expression in the bladder and the effects of 1 h-intravesical infusion of the selective TRPA1 blocker HC-030031 (1 nmol/min) on MGO-induced cystometric alterations. Methods: Five-week-old female C57BL/6 mice received 0.5% MGO in their drinking water for 12 weeks, whereas control mice received tap water alone. Results: Compared to the control group, the protein levels and immunostaining for the MGO-derived hydroimidazolone isomer MG-H1 was increased in bladders of the MGO group, as observed in urothelium and detrusor smooth muscle. TRPA1 protein expression was significantly higher in bladder tissues of MGO compared to control group with TRPA1 immunostaining both lamina propria and urothelium, but not the detrusor smooth muscle. Void spot assays in conscious mice revealed an overactive bladder phenotype in MGO-treated mice characterized by increased number of voids and reduced volume per void. Filling cystometry in anaesthetized animals revealed an increased voiding frequency, reduced bladder capacity, and reduced voided volume in MGO compared to vehicle group, which were all reversed by HC-030031 infusion. Conclusion: TRPA1 activation is implicated in MGO-induced mouse overactive bladder. TRPA1 blockers may be useful to treat diabetic bladder dysfunction in individuals with high MGO levels.

Pyricularia's Capability of Infecting Different Grasses in Two Regions of Mexico.

The genus Pyricularia includes species that are phytopathogenic fungi, which infect different species of Poaceae, such as rice and sorghum. However, few isolates have been genetically characterized in North America. The current study addresses this lack of information by characterizing an additional 57 strains of three grasses (Stenotaphrum secundatum, Cenchrus ciliaris and Digitaria ciliaris) from two distant regions of Mexico. A Pyricularia dataset with ITS sequences retrieved from GenBank and the studied sequences were used to build a haplotype network that allowed us to identify a few redundant haplotypes highly related to P. oryzae species. An analysis considering only the Mexican sequences allowed us to identify non-redundant haplotypes in the isolates of C. ciliaris and D. ciliaris, with a high identity with P. pennisetigena. The Pot2-TIR genomic fingerprinting technique resulted in high variability and allowed for the isolates to be grouped according to their host grass, whilst the ERIC-PCR technique was able to separate the isolates according to their host grass and their region of collection. Representative isolates from different host grasses were chosen to explore the pathogenic potential of these isolates. The selected isolates showed a differential pathogenic profile. Cross-infection with representative isolates from S. secundatum and C. ciliaris showed that these were unable to infect D. ciliaris grass and that the DY1 isolate from D. ciliaris was only able to infect its host grass. The results support the identification of pathogenic strains of Pyricularia isolates and their cross-infection potential in different grasses surrounding important crops in Mexico.

Responsiveness of Candidate Genes on CoPv01CDRK/PhgPv01CDRK Loci in Common Bean Challenged by Anthracnose and Angular Leaf Spot Pathogens.

Anthracnose (ANT) and angular leaf spot (ALS) are significant diseases in common bean, leading to considerable yield losses under specific environmental conditions. The California Dark Red Kidney (CDRK) bean cultivar is known for its resistance to multiple races of both pathogens. Previous studies have identified the CoPv01CDRK/PhgPv01CDRK resistance loci on chromosome Pv01. Here, we evaluated the expression levels of ten candidate genes near the CoPv01CDRK/PhgPv01CDRK loci and plant defense genes using quantitative real-time PCR in CDRK cultivar inoculated with races 73 of Colletotrichum lindemuthianum and 63-39 of Pseudocercospora griseola. Gene expression analysis revealed that the Phvul.001G246300 gene exhibited the most elevated levels, showing remarkable 7.8-fold and 8.5-fold increases for ANT and ALS, respectively. The Phvul.001G246300 gene encodes an abscisic acid (ABA) receptor with pyrabactin resistance, PYR1-like (PYL) protein, which plays a central role in the crosstalk between ABA and jasmonic acid responses. Interestingly, our results also showed that the other defense genes were initially activated. These findings provide critical insights into the molecular mechanisms underlying plant defense against these diseases and could contribute to the development of more effective disease management strategies in the future.

Influence of Tilting Angle on Temperature Measurements of Different Object Sizes Using Fiber-Optic Pyrometers.

This article presents a new model of optical power gathered by a fiber-optic pyrometer when there is a tilting angle between the fiber longitudinal axis and the vector perpendicular to the tangent plane of the emitted surface. This optical power depends on the fiber specifications, such as the diameter and the numerical aperture (NA), as well as the object parameters, including its diameter, emissivity, and tilting angle. Some simulations are carried out using other pyrometers from the literature without tilting to validate the model. Additional simulations with different optical fibers, object sizes, and distances at different tilting angles allow us to describe the behavior of the pyrometer when the object is smaller than the optical fiber field of view (the light cone defined by its NA). The results show that for a finite surface object, the power collected by the optical fiber is affected by changes in the tilting angle, greater tilting lesser gathered power, and reaching the maximum power when the field of view of the fiber covers up the entire object, as expected. On the other hand, additional equations are presented to describe the maximum tilting angle, and distance that allow the maximum power gathered for a determined object diameter and fiber, avoiding temperature measurement errors.

Validation of the use of dried blood spots in a chikungunya virus IgG serological assay.

Dried blood spot (DBS) sampling is a simple, fast, and minimally invasive blood collection method that is particularly useful for diagnostic or epidemiological studies in hard-to-reach populations. Nevertheless, the use of DBS in assays that have been optimized with gold-standard samples (serum or plasma) must be optimized to yield reliable results. Here, we describe the validation of DBS in a commercial assay to measure IgG against chikungunya virus (CHIKV IgG ELISA; Euroimmun, Lübeck, Germany). During a health survey of people experiencing homelessness in Salvador, Brazil, between September 2021 and February 2022, a subset (75/523; 14.3%) of the study participants had paired capillary (for DBS preparation) and venous (for serum separation) blood samples collected. A pilot optimization test was initially performed with 17 paired samples to compare the CHIKV IgG ELISA absorbance values between serum and three different dilutions of DBS. Based on the preliminary results, the best DBS dilution was selected for a final evaluation comparing paired serum and DBS samples from 58 participants. The sensitivity and specificity of the CHIKV ELISA of DBS compared to sera were 100% (95% C.I.: 85.8-100%) and 100% (95% C.I.: 93-100%), respectively. In the linear regression analysis, a coefficient of determination (R2) value of 0.98 indicated the excellent performance of DBS in predicting the serum levels of IgG CHIKV antibodies. Our findings suggest that DBS at an optimized dilution is reliable for investigating the prevalence of CHIKV IgG antibodies during population surveys in the commercial assay tested here.

Characterization of Alternaria alternata and Alternaria scrophulariae Brown Spot in Colombian quinoa (Chenopodium quinoa).

Alternaria is a saprophytic and opportunistic fungus with a worldwide distribution that can affect the quality of various agricultural products, such as fruits, cereals, and pseudocereals. This research was carried out to investigate the population of this genus associated with quinoa cultivation in plots located in the Boyacá department (Colombia), the country's third-largest quinoa-producing department. The present study found 17 Alternaria isolates, of which 13 were identified as A. alternata and 4 as A. scrophulariae (formerly A. conjuncta) employed molecular markers of internal transcribed spacer (ITS) region and translation elongation factor 1α (TEF-1α). In the pathogenicity test under greenhouse conditions, all the Alternaria isolates showed some degree of pathogenicity on Piartal quinoa cultivar plants although no significant differences were found in isolates. The severity indices ranged from 2 to 5, and the percentage of affected leaves per plant ranged between 15% and 40%. This fungus affected the foliar tissue of quinoa, resulting in chlorotic and necrotic spots, symptoms that can generate a reduction in the quality and productivity of crops. This is the first time that the pathogenicity of Alternaria spp. in the Piartal variety has been described and the first report of this genera in quinoa crops of Colombia.

Mapping possible interaction sites for crotoxin in CNF, a gamma PLA2 inhibitor from Crotalus durissus terrificus rattle snake, using SPOT synthesis.

Phospholipases A2 (PLA2s) are main components of snake venoms. Several snake species possess endogenous PLA2 inhibitors in their circulating blood, which are generally known as sbPLIs (an acronym for snake blood phospholipase A2inhibitors). The sbPLIs are categorized in three classes (alpha, beta or gamma) depending on the existence of distinguishing protein domains in their structure. The Crotalus durrissus terrificus venom has a highly neurotoxic PLA2 - crotoxin (CTX) - in its composition and the self-protection of the snake is mainly ensured by a sbγPLI named CNF (standing for Crotalusneutralizing factor). In an attempt to find smaller molecules able to inhibit the catalytic activity of CTX, in the present study we used linear peptide arrays to identify CNF segments possibly involved in the interaction with the toxin. Five reacting segments were identified as possible interacting regions. The target peptides were synthesized and located in the in silico CNF structure. Although all of them are exposed to the solvent, high concentrations were needed to inhibit the PLA2 activity of the whole venom or CTX. Limitations of the methodology employed and particular characteristics of CTX inhibition by CNF are discussed.

The Effects of Gluconacin on Bacterial Tomato Pathogens and Protection against Xanthomonas perforans, the Causal Agent of Bacterial Spot Disease.

As agricultural practices become more sustainable, adopting more sustainable practices will become even more relevant. Searching for alternatives to chemical compounds has been the focus of numerous studies, and bacteriocins are tools with intrinsic biotechnological potential for controlling plant diseases. We continued to explore the biotechnological activity of the bacteriocin Gluconacin from Gluconacetobacter diazotrophicus, PAL5 strain, by investigating this protein's antagonism against important tomato phytopathogens and demonstrating its effectiveness in reducing bacterial spots caused by Xanthomonas perforans. In addition to this pathogen, the bacteriocin Gluconacin demonstrated bactericidal activity in vitro against Ralstonia solanacearum and Pseudomonas syringae pv. tomato, agents that cause bacterial wilt and bacterial spots, respectively. Bacterial spot control tests showed that Gluconacin reduced disease severity by more than 66%, highlighting the biotechnological value of this peptide in ecologically correct formulations.

Evidence that methylglyoxal and receptor for advanced glycation end products are implicated in bladder dysfunction of obese diabetic ob/ob mice.

Glycolytic overload in diabetes causes large accumulation of the highly reactive dicarbonyl compound methylglyoxal (MGO) and overproduction of advanced glycation end products (AGEs), which interact with their receptors (RAGE), leading to diabetes-associated macrovascular complications. The bladder is an organ that stays most in contact with dicarbonyl species, but little is known about the importance of the MGO-AGEs-RAGE pathway to diabetes-associated bladder dysfunction. Here, we aimed to investigate the role of the MGO-AGEs-RAGE pathway in bladder dysfunction of diabetic male and female ob/ob mice compared with wild-type (WT) lean mice. Diabetic ob/ob mice were treated with the AGE breaker alagebrium (ALT-711, 1 mg/kg) for 8 wk in drinking water. Compared with WT animals, male and female ob/ob mice showed marked hyperglycemia and insulin resistance, whereas fluid intake remained unaltered. Levels of total AGEs, MGO-derived hydroimidazolone 1, and RAGE in bladder tissues, as well as fluorescent AGEs in serum, were significantly elevated in ob/ob mice of either sex. Collagen content was also markedly elevated in the bladders of ob/ob mice. Void spot assays in filter paper in conscious mice revealed significant increases in total void volume and volume per void in ob/ob mice with no alterations of spot number. Treatment with ALT-711 significantly reduced the levels of MGO, AGEs, RAGE, and collagen content in ob/ob mice. In addition, ALT-711 treatment normalized the volume per void and increased the number of spots in ob/ob mice. Activation of AGEs-RAGE pathways by MGO in the bladder wall may contribute to the pathogenesis of diabetes-associated bladder dysfunction.NEW & NOTEWORTHY The involvement of methylglyoxal (MGO) and advanced glycation end products (AGEs) in bladder dysfunction of diabetic ob/ob mice treated with the AGE breaker ALT-711 was investigated here. Diabetic mice exhibited high levels of MGO, AGEs, receptor for AGEs (RAGE), and collagen in serum and/or bladder tissues along with increased volume per void, all of which were reduced by ALT-711. Activation of the MGO-AGEs-RAGE pathway in the bladder wall contributes to the pathogenesis of diabetes-associated bladder dysfunction.

Efficacy of ClinproTMXT Varnish surrounding brackets on the enamel surface of white spot lesion

Abstract Orthodontics patients usual develop demineralization and present cavity caries lesions after six months. Minimally invasive procedures have been the goal in modern dental practice. The aim of this study was to evaluate the effect of ClinproTMXT Varnish, on the enamel surface roughness and severity of white spot lesions. Twenty premolars were submitted to bond brackets and experimental induction of demineralization and randomly divided into 2 groups: GI - fluoride varnish (Colgate Duraphat®); GII - Ionomeric Sealant (ClinproTMXT Varnish). The treatment was applied around the brackets. The surface roughness of specimens was analyzed, before treatment and 12 weeks after treatment by laser confocal microscopy, and the severity of the white spot lesion was by laser fluorescence device. The data were analyzed by non-parametric Wilcoxon and Mann-Whitney Test, at 5% significance, roughness percentage reduction was performed. The severity of demineralization decreased in both, GI (p = 0.005) and GII (p = 0.019). Enamel superficial roughness levels decreased in GI and GII. As well as the roughness percentage, being more expressive in the ClinproTMXT Varnish group (85,09%). Colgate Duraphat® or Clinpro™ XT Varnish reduced the severity of the demineralization and decreased the superficial roughness on the enamel. The Clinpro™ XT Varnish was superior to superficial roughness on enamel.

Resumo Pacientes ortodônticos geralmente desenvolvem desmineralização e apresentam lesões de cárie após seis meses de tratamento. Procedimentos minimamente invasivos têm sido o objetivo na prática odontológica moderna. O objetivo deste estudo foi avaliar o efeito do ClinproTM XT selante ionomérico, ao redor do bráquete, com relação a rugosidade superficial do esmalte e a severidade da lesão induzida. Vinte pré-molares foram submetidos a colagem de bráquetes e indução experimental de desmineralização e divididos aleatoriamente em 2 grupos: GI - verniz fluoretado (Colgate Duraphat®); GII - Selante Ionomérico (ClinproTM XT). O tratamento foi aplicado ao redor dos bráquetes. A rugosidade da superfície dos espécimes foi analisada, antes do tratamento e 12 semanas após o tratamento por microscopia confocal a laser e severidade da lesão de mancha branca por dispositivo de fluorescência a laser. Os dados foram analisados pelo teste não paramétrico de Wilcoxon e Mann-Whitney, a 5% de significância. A taxa de redução da lesão foi calculada. A severidade da desmineralização diminuiu tanto no GI (p = 0,005) quanto no GII (p = 0,019). Os níveis de rugosidade superficial do esmalte diminuíram no GI e GII, assim como o percentual de rugosidade, sendo mais expressivo no grupo ClinproTMXT (85,09%). Colgate Duraphat® e Clinpro™ XT reduziram a severidade da desmineralização e diminuíram a rugosidade superficial do esmalte. O selante ionomérico Clinpro™ XT foi superior na redução percentual de rugosidade.