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Canine monocytic ehrlichiosis (CME) is endemic to Brazil, and studies have verified that dogs have been exposed to different genotypes of Ehrlichia canis. This genetic divergence can influence the clinical response of the animals. We aimed to describe clinical and hematological changes in 125 dogs that reacted to BrTRP36, USTRP36, and CRTRP36 genotypes through enzyme immunoassays and to highlight the current concern regarding infection by the Costa Rican genotype. The results showed that 52.0% reacted to the Brazilian genotype, 22.4% reacted to the Costa Rican genotype, and 16.0% reacted to the American genotype, and some co-reactions were observed. Dogs reactive to BrTRP36 were 1.24% more likely to present with medullary regeneration in cases of anemia and 3% less likely to manifest hyperproteinemia, while dogs reactive to CRTRP36 were 0.7% less likely to present with medullary regeneration. Febrile illness and neurological alterations were also statistically associated, with an 85.7% and 231.2% increased likelihood, respectively, to occur in dogs that reacted to USTRP36. The dogs with the American genotype developed clinical manifestations related to systemic inflammation, while those with the Brazilian genotype of E. canis were more dispersed in the region studied, showing greater adaptation to the hosts. We highlight the significant serocurrence of the Costa Rican genotype, which has already been described to have zoonotic potential and which showed less adaptation.

Glomerulonephritis, caused by the deposition of immune complexes, can lead to kidney damage in dogs with canine monocytic ehrlichiosis (CME). The early diagnosis of renal insult is important to prevent severe kidney disease in infected dogs by Ehrlichia canis. This study aimed to investigate urinary biomarkers of renal function, neutrophil gelatinase (uNGAL), and kidney injury molecule-1 (uKIM-1) using the Luminex® xMAP® platform, and the proportion of mixed or high molecular weight proteinuria in dogs with CME. This study included blood samples of thirty dogs with clinical signs of CME and amplified DNA for E. canis (CME group) and six dogs of different breeds and both sexes, aged 3 to 7 years, that showed no clinical-laboratory alterations or tick parasitism and were tested negative for E. canis via PCR (control group). The total calcium, phosphorus (p < 0.05), urea (p < 0.001), creatinine (p < 0.05), urinary density (p < 0.05), urinary protein creatinine ratio (p < 0.001), uNGAL (p < 0.05), and uKIM-1 (p > 0.05), as well as the proportion of high molecular weight proteinuria and mixed proteinuria (p < 0.01), were measured. Elevated serum concentrations of creatinine, urea, and phosphorus combined with reduced urinary density, increased urinary creatinine–protein ratio, urinary NGAL, and mixed proteinuria detected renal damage in dogs with CME, while KIM -1 remained unchanged. uNGAL can detect early renal lesions, reflecting renal damage before a significant increase in serum creatinine occurs, and appears to be an early diagnostic biomarker in renal disease in dogs with CME.

Clinical and hematological characteristics of the blood of 125 dogs with different Ehrlichia canis genotypes were analyzed. We observed that the Brazilian genotype is more prevalent and adapted in dogs in the central-western region of Brazil, whereas the American genotype appears to be more pathogenic, causing inflammatory signs. However, we highlight the high frequency of dogs reactive to the Costa Rican genotype, which is suggested to be less adapted to the immune response of the animals. In addition, this genotype has an imminent risk because of its zoonotic potential. Canine monocytic ehrlichiosis (CME) is endemic to Brazil, and studies have verified that dogs have been exposed to different genotypes of Ehrlichia canis. This genetic divergence can influence the clinical response of the animals. We aimed to describe clinical and hematological changes in 125 dogs that reacted to BrTRP36, USTRP36, and CRTRP36 genotypes through enzyme immunoassays and to highlight the current concern regarding infection by the Costa Rican genotype. The results showed that 52.0% reacted to the Brazilian genotype, 22.4% reacted to the Costa Rican genotype, and 16.0% reacted to the American genotype, and some co-reactions were observed. Dogs reactive to BrTRP36 were 1.24% more likely to present with medullary regeneration in cases of anemia and 3% less likely to manifest hyperproteinemia, while dogs reactive to CRTRP36 were 0.7% less likely to present with medullary regeneration. Febrile illness and neurological alterations were also statistically associated, with an 85.7% and 231.2% increased likelihood, respectively, to occur in dogs that reacted to USTRP36. The dogs with the American genotype developed clinical manifestations related to systemic inflammation, while those with the Brazilian genotype of E. canis were more dispersed in the region studied, showing greater adaptation to the hosts. We highlight the significant serocurrence of the Costa Rican genotype, which has already been described to have zoonotic potential and which showed less adaptation.

Background The continuous proliferation of intestinal stem cells followed by their tightly regulated differentiation to epithelial cells is essential for the maintenance of the gut epithelial barrier and its functions. How these processes are tuned by diet and gut microbiome is an important, but poorly understood question. Dietary soluble fibers, such as inulin, are known for their ability to impact the gut bacterial community and gut epithelium, and their consumption has been usually associated with health improvement in mice and humans. In this study, we tested the hypothesis that inulin consumption modifies the composition of colonic bacteria and this impacts intestinal stem cells functions, thus affecting the epithelial structure. Methods Mice were fed with a diet containing 5% of the insoluble fiber cellulose or the same diet enriched with an additional 10% of inulin. Using a combination of histochemistry, host cell transcriptomics, 16S microbiome analysis, germ-free, gnotobiotic, and genetically modified mouse models, we analyzed the impact of inulin intake on the colonic epithelium, intestinal bacteria, and the local immune compartment. Results We show that the consumption of inulin diet alters the colon epithelium by increasing the proliferation of intestinal stem cells, leading to deeper crypts and longer colons. This effect was dependent on the inulin-altered gut microbiota, as no modulations were observed in animals deprived of microbiota, nor in mice fed cellulose-enriched diets. We also describe the pivotal role of γδ T lymphocytes and IL-22 in this microenvironment, as the inulin diet failed to induce epithelium remodeling in mice lacking this T cell population or cytokine, highlighting their importance in the diet-microbiota-epithelium-immune system crosstalk. Conclusion This study indicates that the intake of inulin affects the activity of intestinal stem cells and drives a homeostatic remodeling of the colon epithelium, an effect that requires the gut microbiota, γδ T cells, and the presence of IL-22. Our study indicates complex cross kingdom and cross cell type interactions involved in the adaptation of the colon epithelium to the luminal environment in steady state. -dZ131ovLvfeoRbZ-Pz4qe Video Abstract

Background Lipid nanoparticles (LNP) are a safe and effective messenger RNA (mRNA) delivery system for vaccine applications, as shown by the COVID-19 mRNA vaccines. One of the main challenges faced during the development of these vaccines is the production of new and versatile LNP formulations capable of efficient encapsulation and delivery to cells in vivo. This study aimed to develop a new mRNA vaccine formulation that could potentially be used against existing diseases as well as those caused by pathogens that emerge every year. Results Using firefly luciferase (Luc) as a reporter mRNA, we evaluated the physical–chemical properties, stability, and biodistribution of an LNP-mRNA formulation produced using a novel lipid composition and a microfluidic organic-aqueous precipitation method. Using mRNAs encoding a dengue virus or a Leishmania infantum antigen, we evaluated the immunogenicity of LNP-mRNA formulations and compared them with the immunization with the corresponding recombinant protein or plasmid-encoded antigens. For all tested LNP-mRNAs, mRNA encapsulation efficiency was higher than 85%, their diameter was around 100 nm, and their polydispersity index was less than 0.3. Following an intramuscular injection of 10 µg of the LNP-Luc formulation in mice, we detected luciferase activity in the injection site, as well as in the liver and spleen, as early as 6 h post-administration. LNPs containing mRNA encoding virus and parasite antigens were highly immunogenic, as shown by levels of antigen-specific IgG antibody as well as IFN-γ production by splenocytes of immunized animals that were similar to the levels that resulted from immunization with the corresponding recombinant protein or plasmid DNA. Conclusions Altogether, these results indicate that these novel LNP-mRNA formulations are highly immunogenic and may be used as novel vaccine candidates for different infectious diseases. Graphical Abstract

The Cactaceae family is composed by 124 genera and about 1438 species. Pilosocereus gounellei, popularly known in Brazil as xique-xique, is used in folk medicine to treat prostate inflammation, gastrointestinal and urinary diseases. The pioneering phytochemical study of P. gounellei was performed using column chromatography and HPLC, resulting in the isolation of 10 substances: pinostrobin (1), β-sitosterol (2), a mixture of sitosterol 3-O-β-d-glucopyranoside/stigmasterol 3-O-β-d-glucopyranoside (3a/3b), 132-hydroxyphaeophytin a (4), phaeophytin a (5), a mixture of β-sitosterol and stigmasterol (6a/6b), kaempferol (7), quercetin (8), 7′-ethoxy-trans-feruloyltyramine (mariannein, 9) and trans-feruloyl tyramine (10). Compound 9 is reported for the first time in the literature. The structural characterization of the compounds was performed by analyses of 1-D and 2-D NMR data. In addition, a phenolic and flavonol total content assay was carried out, and the anti-oxidant potential of P. gounellei was demonstrated.

ABSTRACT Remote monitoring of the management of coffee crops is necessary as the demand in decision-making, where the aim is to rise production based on sustainable management is in a constant growth. In this work, it was evaluated the potential of images obtained by low-cost sensors in the discrimination of sources and doses of mineral and organomineral fertilizers in coffee. The experimental design was in randomized blocks, with five blocks and six treatments, as follows: (T1) - 100% of the organomineral treatment; (T2) - 70% of the organomineral treatment; (T3) - 50% of the organomineral treatment; (T4) - 100% of mineral fertilization; (T5) - standard treatment of the farm and (T6) - 70% of mineral fertilization. After management, we used the Mapir 3 Survey3W camera coupled to an ARP drone – Phantom4 to take images of the experiment over a 12-month vegetative period. Combined with image taking, it was collected agronomic parameters of coffee growth and productivity for two crops and concluded that different fertilization doses did not significantly affect the analyzed parameters. Based on the supervised classification of multispectral images, it was possible to discriminate treatments with a higher degree of accuracy (86.66% accuracy) than when analyzing coffee growth parameters.

Cats naturally exposed to Ehrlichia canis have been described in different regions of the world, but little is known about the genotypes associated with infection in these animals. To detect E. canis-specific antibodies and investigate the E. canis TRP genotypes in cats, serum samples from 76 domestic cats reactive to crude E. canis antigens by the indirect fluorescence antibody test (IFAT) were analyzed by ELISA, using E. canis-specific peptides (i.e., TRP19 and TRP36 /BR/US/CR). Of these, 25 (32.9%) cats reacted to at least one TRP peptide, confirming their specific exposure to E. canis. Eighteen (23.7%) cats reacted to TRP19, 15 (19.8%) to BRTRP36, and 11 (14.5%) to USTRP36, but none of them reacted to CRTRP36. Eight (10.5%) cats reacted to TRP19 but not to any TRP36 genotype, demonstrating the possible existence of a new E. canis genotype infecting felines. Nevertheless, this study provides the first report of anti-E. canis-specific antibodies in domestic cats.

Canine distemper virus (CDV) is a serious and often fatal disease caused by Morbillivirus canis, which affects domestic dogs and wild carnivores, with case-fatality rates reaching up to 47%. The hemagglutinin (H) protein mediates viral adsorption and shows high genetic variability, making it a valuable molecular marker. This study aimed to detect and characterize the H gene of CDV strains from 14 dogs with fatal neurological disease in the Brazilian states of Mato Grosso and Rondônia. Brain tissue was tested via RT-PCR for the nucleocapsid gene, and positive samples were amplified for the H gene. Ten complete H-gene sequences were obtained. Phylogenetic analysis revealed two distinct clusters within the South America I/Europe lineage: one related to strains from Uruguay and Argentina (with residues 530G/549Y) and another related to Brazilian strains (530S/549Y). One sequence (MT8) showed an intermediate position in the haplotype network but clustered phylogenetically with Uruguay/Argentina-related strains. Most sequences carried 530S/549Y, a pattern linked to altered SLAM receptor usage in wildlife. These findings demonstrate the co-circulation of two CDV clusters in Central–Western Brazil, their regional and international genetic connectivity, and amino acid substitutions potentially influencing host adaptation and antigenicity.

ObjectiveThis study aims to retrospectively analyze anatomical parameters for safe LCP performance and provide a technical note on its execution.MethodsWe retrospectively analyzed 200 head computed tomography (CT) scans to identify the optimal LCP point (P), defined by the intersection of the anterior margin of the C1 posterior arch and the midpoint between the inferior C1 posterior arch and superior C2 lamina. Anatomical distances (D1, D2, D3) from this point to relevant landmarks were measured. Statistical analyses were performed using the Mann-Whitney U test and Student’s t-test to compare groups. A detailed technical note on LCP execution, including patient positioning, anatomical considerations, and procedural steps, is provided, augmented by an illustrative case.ResultsThe mean age of the 200 patients (108 females, 92 males) was 63.67 ± 11.79 years. Mean distances were D1: 7.04 ± 2.09 mm, D2: 14.59 ± 4.94 mm, and D3: 12.17 ± 4.79 mm. No statistically significant differences in puncture distances were observed between sexes or age groups (≤ 60 vs. >60 years), except for D3 in women, where those over 60 years exhibited a significantly greater mean distance compared to younger women (p = 0.047). The illustrative case describes a 61-year-old male who developed a surgical site infection and required CSF analysis, for which LCP was successfully performed due to contraindication for LP, yielding clear CSF with negative cultures.ConclusionsLCP is a safe and effective alternative for CSF collection in scenarios where LP is contraindicated. Our anatomical findings, supported by the illustrative case, delineate consistent landmarks and suggest minimal anatomical variation that might necessitate slight adjustments, particularly for D3 in older women. The detailed technical approach combined with anatomical precision enhances the safety and efficacy of LCP, making it a valuable tool in neurosurgical practice.