Periparturient blood T-lymphocyte PD-1 and CTLA-4 expression as potential predictors of new intramammary infections in dairy cows during early lactation (short communication).

BACKGROUND: The periparturient period in dairy cows is marked by immunosuppression which increases the likelihood of infectious disorders, particularly also mastitis. An in-depth understanding of peripartum leukocyte biology is vital for the implementation of highly successful post-partum disease prevention measures. Immune checkpoint molecules, such as programmed death 1 (PD-1) and cytotoxic T-lymphocyte antigen 4 (CTLA-4), are critical inhibitory receptors expressed on immune cells, particularly T cells, that drive immunosuppressive signaling pathways. However, the potential role of immune checkpoint molecules expression in T-cells on udder health has never been explored. Thus, the association between the occurrence of new postpartum intramammary infections (IMIs) and the expression of programmed cell death protein 1 (PD-1) and cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) on blood T-cells during the peripartum period was investigated. RESULTS: In this study, the incidence of IMIs by any pathogen in early lactation was not associated with a higher expression of PD-1 and CTLA-4 in the periparturient period. However, the incidence of IMIs by major pathogens throughout the first month of lactation was significantly associated with higher expression of PD-1 at 14 days before calving (P = 0.03) and CTLA-4 at parturition (P = 0.03) by blood T-cells. Also, the expression of CTLA-4 at D0 (P = 0.012) by T-cells was associated with the occurrence of persistent IMIs during the first month of lactation. CONCLUSIONS: To our knowledge, this is the first report to investigate the expression of PD-1 and CTLA-4 by blood T-lymphocytes during the periparturient period in dairy cows and to explore their relationship with the incidence of new IMIs in the postpartum period. Thus, a comprehensive understanding of leukocyte biology during peripartum would appear to be a prerequisite for the identification of resilient dairy cows or targets innovative (immunological) non-antibiotic approaches in the transition period.

Thiazolyl-isatin derivatives: Synthesis, in silico studies, in vitro biological profile against breast cancer cells, mRNA expression, P-gp modulation, and interactions of Akt2 and VIM proteins.

The literature reports that thiazole and isatin nuclei present a range of biological activities, with an emphasis on anticancer activity. Therefore, our proposal was to make a series of compounds using the molecular hybridization strategy, which has been used by our research group, producing hybrid molecules containing the thiazole and isatin nuclei. After structural planning and synthesis, the compounds were characterized and evaluated in vitro against breast cancer cell lines (T-47D, MCF-7 and MDA-MB-231) and against normal cells (PBMC). The activity profile on membrane proteins involved in chemoresistance and tumorigenic signaling proteins was also evaluated. Among the compounds tested, the compounds 4c and 4a stood out with IC50 values of 1.23 and 1.39 µM, respectively, against the MDA-MB-231 cell line. Both compounds exhibited IC50 values of 0.45 µM for the MCF-7 cell line. Compounds 4a and 4c significantly decreased P-gp mRNA expression levels in MCF-7, 4 and 2 folds respectively. Regarding the impact on tumorigenic signaling proteins, compound 4a inhibited Akt2 in MDA-MB-231 and compound 4c inhibited the mRNA expression of VIM in MCF-7.

Disease progression in Sanfilippo type B: Case series of Brazilian patients.

Mucopolysaccharidosis type IIIB (MPS IIIB) is caused by deficiency of alpha-N-acetylglucosaminidase, leading to storage of heparan sulphate. The disease is characterized by intellectual disability and hyperactivity, among other neurological and somatic features. Here we studied retrospective data from a total of 19 MPS IIIB patients from Brazil, aiming to evaluate disease progression. Mean age at diagnosis was 7.2 years. Speech delay was one of the first symptoms to be identified, around 2-3 years of age. Behavioral alterations include hyperactivity and aggressiveness, starting around age four. By the end of the first decade, patients lost acquired abilities such as speech and ability to walk. Furthermore, as disease progresses, respiratory, cardiovascular and joint abnormalities were found in more than 50% of the patients, along with organomegaly. Most common cause of death was respiratory problems. The disease progression was characterized in multiple systems, and hopefully these data will help the design of appropriate clinical trials and clinical management guidelines.

Herbicide Leaching in Soil with Different Properties: Perspectives from Commercial Formulations and Analytical Standards.

The leaching of herbicides into the soil is essential to control germinating seeds and parts of vegetative weeds. However, herbicide transportation to deeper soil layers can result in groundwater contamination and, consequently, environmental issues. In this research, our objective was to investigate differences in herbicide leaching between commercial formulations and analytical standards using three different soils. Leaching experiments were carried out for diuron, hexazinone, and sulfometuron-methyl herbicides isolated and in binary and ternary mixtures. The herbicide residue quantification was performed by ultra-high-performance liquid chromatography coupled to a mass spectrometer (LC-MS/MS). Diuron had less mobility in soils and was retained in the most superficial layers. Hexazinone and sulfometuron-methyl were more mobile and leached into deeper layers. The leaching process was more intense for hexazinone and sulfometuron-methyl. The additives present in the commercial formulation favored the leaching in soils of diuron, hexazinone, and sulfometuron-methyl herbicides isolated and mixture compared to the analytical standard. This fact highlights the importance of considering these effects for the positioning of herbicides in the field to increase the efficiency of weed control and minimize the potential for environmental contamination.

Effect of Ceramic Thickness on the Bond Strength to Resin-Luting Agents before and after Thermal Cycling.

This study investigated microshear bond strength (µSBS) of two (2) dual-cured resin-luting agents (RelyX™ Ultimate and RelyX™ U200) when photoactivated through varying thicknesses of lithium disilicate, with or without thermal cycling. Discs of IPS e.max Press of 0.5, 1.5, and 2 mm in thickness were obtained. Elastomer molds (3.0 mm in thickness) with four cylinder-shaped orifices 1.0 mm in diameter, were placed onto the ceramic surfaces and filled with resin-luting agents. A Mylar strip, glass plate, and load of 250 grams were placed over the filled mold. The load was removed and the resin-luting agents were photoactivated through the ceramics using a single-peak LED (Radii Plus.) All samples were stored in distilled water at 37oC for 24 h. Half of the samples were subjected to thermal cycling (3,000 cycles; 5ºC and 55ºC). All samples were then submitted to µSBS test using a universal testing machine (Instron 4411) at a crosshead speed of 0.5 mm/min. Data were submitted to three-way ANOVA and Tukey post-hoc test (α=0.05). The mean µSBS at 24 h was significantly higher than after thermal cycling (p<0.05). No statistical difference was found between resin-luting agents (p > 0.05). The mean µSBS for groups photoactivated through 0.5 mm ceramic were significantly higher than 1.5 mm and 2.0 mm (p < 0.05). In conclusion, increased ceramic thicknesses reduced the bond strength of tested resin-luting agents to lithium disilicate. No differences were found between resin-luting agents. Thermal cycling reduced the bond strength of both resin-luting agents.

Nanoencapsulation of hybrid crude palm oil Unaué HIE OxG with jackfruit by-products as encapsulants: A study of cellular antioxidant activity and cytotoxicity in Caco-2 cells.

Hybrid crude palm oil (HCPO) HIE OxG is notable for its abundance of carotenoids, tocopherols, and tocotrienols. Investigating cellular antioxidant activity (CAA) and the non-cytotoxicity of oil nanoparticles is crucial for understanding the behavior of these phytochemicals in biological systems and ensuring the safety of products. Nanoparticles of HCPO, encapsulated with jackfruit by-products were produced and characterized for CAA and cytotoxicity in Caco-2 cells. The nanoparticles exhibited nanoscale diameters (<250 nm), uniform distribution and stability (polydispersity index < 0.25; zeta potential JSF-NP -12.46 ± 0.15 mV and JAF-NP -13.73 ± 1.28 mV). JSF-NP and JAF-NP demonstrated superior CAA compared to the free HCPO across all concentrations, without inducing cytotoxic effects on differentiated Caco-2 cells. This study underscores the importance of investigating the CAA of edible oil nanoparticles, with non-cytotoxicity indicating biological safety and the potential to safeguard intestinal epithelial cells. Thus, JSF-NP and JAF-NP emerge as promising delivery systems for future HCPO applications.

Classic and Atypical Late Infantile Neuronal Ceroid Lipofuscinosis in Latin America: Clinical and Genetic Aspects, and Treatment Outcome with Cerliponase Alfa.

Introduction: Late infantile neuronal ceroid lipofuscinosis type 2 (CLN2), is a neurodegenerative autosomal recessive disease caused by TPP1 gene variants, with a spectrum of classic and atypical phenotypes. The aim of treatment is to slow functional decline as early as possible in an attempt to improve quality of life and survival. This study describes the clinical characteristics as well as the response to treatment with cerliponase alfa. Materials and methods: A retrospective study was conducted in five Latin-American countries, using clinical records from patients with CLN2. Clinical follow-up and treatment variables are described. A descriptive and bivariate statistical analysis was performed. Results: A total of 36 patients were observed (range of follow-up of 61-110 weeks post-treatment). At presentation, patients with the classic phenotype (n = 16) exhibited regression in language (90%), while seizures were the predominant symptom (87%) in patients with the atypical phenotype (n = 20). Median age of symptom onset and time to first specialized consultation was 3 (classical) and 7 (atypical) years, while the median time interval between onset of symptoms and treatment initiation was 4 years (classical) and 7.5 (atypical). The most frequent variant was c.827 A > T in 17/72 alleles, followed by c.622C > T in 6/72 alleles. All patients were treated with cerliponase alfa, and either remained functionally stable or had a loss of 1 point on the CLN2 scale, or up to 2 points on the Wells Cornel and Hamburg scales, when compared to pretreatment values. Discussion and conclusion: This study reports the largest number of patients with CLN2 currently on treatment with cerliponase alfa in the world. Data show a higher frequency of patients with atypical phenotypes and a high allelic proportion of intron variants in our region. There was evidence of long intervals until first specialized consultation, diagnosis, and enzyme replacement therapy. Follow-up after the initiation of cerliponase alfa showed slower progression or stabilization of the disease, associated with adequate clinical outcomes and stable functional scores. These improvements were consistent in both clinical phenotypes.

Optimization and Characterization of Interspecific Hybrid Crude Palm Oil Unaué HIE OxG Nanoparticles with Vegetable By-Products as Encapsulants.

Interspecific hybrid crude palm oil (HCPO) HIE OxG derived from crossbred African oil palm (Elaeis guineensis) and American Caiaué (Elaeis oleifera) is prominent for its fatty acid and antioxidant compositions (carotenoids, tocopherols, and tocotrienols), lower production cost, and high pest resistance properties compared to crude palm oil. Biodegradable and sustainable encapsulants derived from vegetable byproducts were used to formulate HCPO nanoparticles. Nanoparticles with hybrid crude palm oil and jackfruit seed flour as a wall material (N-JSF) and with hybrid crude palm oil and jackfruit axis flour as a wall material (N-JAF) were optimized using a 22 experimental design. They exhibited nanoscale diameters (<250 nm) and were characterized based on their zeta potential, apparent viscosity, pH, color, and total carotenoid content. The nanoparticles demonstrated a monodisperse distribution, good uniformity, and stability (polydispersity index < 0.25; zeta potentials: N-JSF -19.50 ± 1.47 mV and N-JAF -12.50 ± 0.17 mV), as well as high encapsulation efficiency (%) (N-JSF 86.44 ± 0.01 and N-JAF 90.43 ± 1.34) and an optimal carotenoid retention (>85%). These nanoparticles show potential for use as sustainable and clean-label HCPO alternatives in the food industry.

Acute Dyspnea With an Infiltrative Tracheal Mass.

CASE PRESENTATION: A 48-year-old woman presented to the ED with a nonproductive cough, shortness of breath, and stridor. She was otherwise healthy and had never used tobacco. The patient was mildly tachycardic but otherwise hemodynamically stable, afebrile, and saturating well on room air. She did not display any signs of increased work of breathing at rest. Although auscultation of her thorax indicated good air entry bilaterally without any adventitious sounds, stridor was elicited with forced expiration.

Clinical Characterization and Underlying Genetic Findings in Brazilian Patients with Syndromic Microcephaly Associated with Neurodevelopmental Disorders.

Microcephaly is characterized by an occipitofrontal circumference at least two standard deviations below the mean for age and sex. Neurodevelopmental disorders (NDD) are commonly associated with microcephaly, due to perturbations in brain development and functioning. Given the extensive genetic heterogeneity of microcephaly, managing patients is hindered by the broad spectrum of diagnostic possibilities that exist before conducting molecular testing. We investigated the genetic basis of syndromic microcephaly accompanied by NDD in a Brazilian cohort of 45 individuals and characterized associated clinical features, as well as evaluated the effectiveness of whole-exome sequencing (WES) as a diagnostic tool for this condition. Patients previously negative for pathogenic copy number variants underwent WES, which was performed using a trio approach for isolated index cases (n = 31), only the index in isolated cases with parental consanguinity (n = 8) or affected siblings in familial cases (n = 3). Pathogenic/likely pathogenic variants were identified in 19 families (18 genes) with a diagnostic yield of approximately 45%. Nearly 86% of the individuals had global developmental delay/intellectual disability and 51% presented with behavioral disturbances. Additional frequent clinical features included facial dysmorphisms (80%), brain malformations (67%), musculoskeletal (71%) or cardiovascular (47%) defects, and short stature (54%). Our findings unraveled the underlying genetic basis of microcephaly in half of the patients, demonstrating a high diagnostic yield of WES for microcephaly and reinforcing its genetic heterogeneity. We expanded the phenotypic spectrum associated with the condition and identified a potentially novel gene (CCDC17) for congenital microcephaly.

Body image perception and dissatisfaction in resistance training practitioners: A critical review of the literature of Brazilian studies and assessment tools used.

This study aimed to review the findings on body image (BI) dissatisfaction and muscle dysmorphia (MD) in Brazilian RT practitioners and the differences in the assessment tools used. A critical review of studies with searches in PubMed, Brazilian Virtual Health Library, SciELO, PsycInfo, and SPORTDiscus databases was conducted. A total of 23 studies were included. Nine tools were used to assess BI dissatisfaction or MD: three questionnaires and six visual scales. The overall mean BI dissatisfaction was 56.5% (59.2% in men vs 57.3% in women). The mean MD was 42.4% (45.1% in women vs 38.5% in men). BI dissatisfaction and MD are related to women seeking slimness and men wanting to increase muscle mass. In conclusion, the frequency of BI was high in both sexes, and MD, when identified, was higher in women. The scales and questionnaires used differ significantly in depth and scope for the same purpose.

Development of ternary polymeric films based on cassava starch, pea flour and green banana flour for food packaging.

The development of alternative materials to replace plastics used in food packaging is an important approach to reducing environmental pollution and minimizing harmful impacts on ecosystems. In this study, biopolymeric films were formulated using cassava starch (Manihot esculenta Crantz), pea flour (Pisum sativum) and green banana flour (Musa sp.) to obtain a material for application in food packaging. The influence of a plasticizer on the optical and physicochemical properties of the films was analyzed and the synergy between higher concentrations of starch and plasticizer resulted in films with low opacity. In addition, the morphology, thermal, mechanical and barrier properties were examined. The film with the best formulation (p < 0.05) contained 12 g cassava starch, 3.6 g pea flour and 30 % glycerol (the maximum levels of the experiment). This film presented average values of thickness, moisture, solubility, opacity, maximum strength (F), maximum tensile stress (σ), elongation at break (ε) and elasticity (E) of 0.47 mm, 19.95 %, 87.45 %, 20.93 %, 9.30 N, 1.75 MPa, 30.10 % and 5.93 %, respectively. This research demonstrates the potential application of films obtained by combining starches from different sources. The sustainable production of environmentally-friendly packaging provides an alternative to fossil-based plastics, which have well-documented adverse effects on the environment.

Insect residues as an alternative and promising source for the extraction of chitin and chitosan.

This work aimed to obtain and characterize chitin and chitosan extracted from the rearing residues of Tenebrio molitor, Zophobas morio, and Blaptica dubia insects in different growth stages in the same rearing cycles chitin and chitosan yielded 11.21 %-20.89 % and 6.26 %-7.07 %, respectively. The deacetylation degrees of chitosan ranged from 75.75 %-89.21 %, and the solubilities from 69.88 %-94.39 %. Infrared spectroscopy corroborated the acquisition of chitin and chitosan and can be used as a semi-quantitative technique for determining the degree of chitosan deacetylation. The X-ray diffraction profiles revealed the presence of α-chitin, and the relative crystalline indices ranged from 65.9 %-89.2 %. Typical TG profiles with two thermal events are observed for chitin and chitosan samples with different residue contents from the extraction procedure. The chitosan solutions exhibited pseudoplastic behavior, with apparent viscosities ranging from 195.96 to 249.86 mPa.s. The characterization results of the biopolymers extracted from insect residues were similar to those obtained from conventional sources. The growth stage influenced the chitin yield and crystallinity index. The results of this study reinforce the feasibility of using alternative sources of chitin and chitosan, providing the use of waste from insect farms and contributing to sustainability and a circular economy.

Effect of Ceramic Thickness on the Bond Strength to Resin-Luting Agents before and after Thermal Cycling

Abstract This study investigated microshear bond strength (µSBS) of two (2) dual-cured resin-luting agents (RelyX™ Ultimate and RelyX™ U200) when photoactivated through varying thicknesses of lithium disilicate, with or without thermal cycling. Discs of IPS e.max Press of 0.5, 1.5, and 2 mm in thickness were obtained. Elastomer molds (3.0 mm in thickness) with four cylinder-shaped orifices 1.0 mm in diameter, were placed onto the ceramic surfaces and filled with resin-luting agents. A Mylar strip, glass plate, and load of 250 grams were placed over the filled mold. The load was removed and the resin-luting agents were photoactivated through the ceramics using a single-peak LED (Radii Plus.) All samples were stored in distilled water at 37oC for 24 h. Half of the samples were subjected to thermal cycling (3,000 cycles; 5ºC and 55ºC). All samples were then submitted to µSBS test using a universal testing machine (Instron 4411) at a crosshead speed of 0.5 mm/min. Data were submitted to three-way ANOVA and Tukey post-hoc test (α=0.05). The mean µSBS at 24 h was significantly higher than after thermal cycling (p<0.05). No statistical difference was found between resin-luting agents (p > 0.05). The mean µSBS for groups photoactivated through 0.5 mm ceramic were significantly higher than 1.5 mm and 2.0 mm (p < 0.05). In conclusion, increased ceramic thicknesses reduced the bond strength of tested resin-luting agents to lithium disilicate. No differences were found between resin-luting agents. Thermal cycling reduced the bond strength of both resin-luting agents.

Resumo: Este estudo investigou a resistência de união ao microcisalhamento (RUµC) de dois (2) agentes de cimentação de resina dual (RelyX™ Ultimate e RelyX™ U200) quando fotoativados através de diferentes espessuras de dissilicato de lítio, com ou sem ciclagem térmica. Discos do IPS e.max Press de 0,5, 1,5 e 2 mm de espessura foram obtidos. Moldes de elastômero (3,0 mm de espessura) com quatro orifícios cilíndricos de 1,0 mm de diâmetro foram colocados sobre as superfícies cerâmicas e preenchidos com agentes de cimentação de resina. Uma tira Mylar, placa de vidro e carga de 250 gramas foram colocadas sobre o molde preenchido. A carga foi removida e os agentes de cimentação resinosos foram fotoativados através da cerâmica usando um LED de pico-único (Radii Plus). Todas as amostras foram armazenadas em água deionizada a 37oC por 24 h. Metade das amostras foi submetida a ciclagem térmica (3.000 ciclos; 5ºC e 55ºC). Todas as amostras foram então submetidas ao teste de RUµC usando uma máquina de teste universal (Instron 4411) com velocidade de 0,5 mm/min. Os dados foram submetidos à Análise de Variância três fatores e ao teste post-hoc de Tukey (α = 0,05). A média de RUµC em 24 h foi significativamente maior do que após a ciclagem térmica (p < 0,05). Não houve diferença estatística entre os cimentos resinosos (p > 0,05). As médias de RUµC para grupos fotoativados através de cerâmica de 0,5 mm foram significativamente maiores do que 1,5 mm e 2,0 mm (p < 0,05). Em conclusão, o aumento da espessura da cerâmica reduziu a resistência de união dos agentes de cimentação resinosos ao dissilicato de lítio. Não foram encontradas diferenças entre os agentes de cimentação resinosos. A ciclagem térmica reduziu a resistência de união de ambos os agentes de cimentação resinosos.

Enhancing Anthocyanin Extraction from Wine Lees: A Comprehensive Ultrasound-Assisted Optimization Study.

Wine lees, an important by-product of the wine industry, pose a major environmental problem due to the enormous quantities of solid-liquid waste that are discarded annually without defined applications. In this study, the optimization of a method based on a Box-Behnken design with surface response has been carried out to obtain extracts with high anthocyanin content and potent antioxidant activity. Six variables have been considered: %EtOH, temperature, amplitude, cycle, pH, and ratio. The developed method exhibited important repeatability properties and intermediate precision, with less than 5% CV being achieved. Furthermore, these novel methods were successfully applied to diverse wine lees samples sourced from Cabernet Sauvignon and Syrah varieties (Vitis vinifera), resulting in extracts enriched with significant anthocyanin content and noteworthy antioxidant activity. Additionally, this study evaluated the influence of grape variety, fermentation type (alcoholic or malolactic), and sample treatment on anthocyanin content and antioxidant activity, providing valuable insights for further research and application in various sectors. The potential applications of these high-quality extracts extend beyond the winemaking industry, holding promise for fields like medicine, pharmaceuticals, and nutraceuticals, thus promoting a circular economy and mitigating environmental contamination.