Post-translational modification by the Pgf glycosylation machinery modulates Streptococcus mutans OMZ175 physiology and virulence.

Streptococcus mutans is commonly associated with dental caries and the ability to form biofilms is essential for its pathogenicity. We recently identified the Pgf glycosylation machinery of S. mutans, responsible for the post-translational modification of the surface-associated adhesins Cnm and WapA. Since the four-gene pgf operon (pgfS-pgfM1-pgfE-pgfM2) is part of the S. mutans core genome, we hypothesized that the scope of the Pgf system goes beyond Cnm and WapA glycosylation. In silico analyses and tunicamycin sensitivity assays suggested a functional overlap between the Pgf machinery and the rhamnose-glucose polysaccharide synthesis pathway. Phenotypic characterization of pgf mutants (ΔpgfS, ΔpgfE, ΔpgfM1, ΔpgfM2, and Δpgf) revealed that the Pgf system is important for biofilm formation, surface charge, membrane stability, and survival in human saliva. Moreover, deletion of the entire pgf operon (Δpgf strain) resulted in significantly impaired colonization in a rat oral colonization model. Using Cnm as a model, we showed that Cnm is heavily modified with N-acetyl hexosamines but it becomes heavily phosphorylated with the inactivation of the PgfS glycosyltransferase, suggesting a crosstalk between these two post-translational modification mechanisms. Our results revealed that the Pgf machinery contributes to multiple aspects of S. mutans pathobiology that may go beyond Cnm and WapA glycosylation.

Experimental Periodontitis Worsens Dopaminergic Neuronal Degeneration.

AIM: To investigate the hypothesis supporting the link between periodontitis and dopaminergic neuron degeneration. MATERIALS AND METHODS: Adult male Wistar rats were used to induce dopaminergic neuronal injury with 6-hydroxydopamine (6-OHDA) neurotoxin and experimental periodontitis via ligature placement. Motor function assessments were conducted before and after periodontitis induction in controls and 6-OHDA-injury-induced rats. Tissue samples from the striatum, jaw and blood were collected for molecular analyses, encompassing immunohistochemistry of tyrosine hydroxylase, microglia and astrocyte, as well as micro-computed tomography, to assess alveolar bone loss and for the analysis of striatal oxidative stress and plasma inflammatory markers. RESULTS: The results indicated motor impairment in 6-OHDA-injury-induced rats exacerbated by periodontitis, worsening dopaminergic striatal degeneration. Periodontitis alone or in combination with 6-OHDA-induced lesion was able to increase striatal microglia, while astrocytes were increased by the combination only. Periodontitis increased striatal reactive oxygen species levels and plasma tumour necrosis factor-alpha levels in rats with 6-OHDA-induced lesions and decreased the anti-inflammatory interleukin-10. CONCLUSIONS: This study provides original insights into the association between periodontitis and a neurodegenerative condition. The increased inflammatory pathway associated with both 6-OHDA-induced dopaminergic neuron lesion and periodontal inflammatory processes corroborates that the periodontitis-induced systemic inflammation may aggravate neuroinflammation in Parkinson's-like disease, potentially hastening disease progression.

Enterotoxigenic Staphylococcus aureus in Brazilian artisanal cheeses: Occurrence, counts, phenotypic and genotypic profiles.

The present study aimed to assess the occurrence and counts of Staphylococcus aureus in Brazilian artisanal cheeses (BAC) produced in five regions of Brazil: Coalho and Manteiga (Northeast region); Colonial and Serrano (South); Caipira (Central-West); Marajó (North); and Minas Artisanal cheeses, from Araxá, Campos das Vertentes, Cerrado, Serro and Canastra microregions (Southeast). The resistance to chlorine-based sanitizers, ability to attach to stainless steel surfaces, and antibiogram profile of a large set of S. aureus strains (n = 585) were assessed. Further, a total of 42 isolates were evaluated for the presence of enterotoxigenic genes (sea, seb, sec, sed, see, seg, sei, sej, and ser) and submitted to typing using pulsed-field gel electrophoresis (PFGE). BAC presented high counts of S. aureus (3.4-6.4 log CFU/g), varying from 25 to 62.5%. From the S. aureus strains (n = 585) assessed, 16% could resist 200 ppm of sodium hypochlorite, whereas 87.6% produced strong ability to attach to stainless steel surfaces, corroborating with S. aureus ability to persist and spread in the environment. Furthermore, the relatively high frequency (80.5%) of multidrug-resistant S. aureus and the presence of enterotoxin genes in 92.6% of the strains is of utmost attention. It reveals the lurking threat of SFP that can survive when conditions are favorable. The presence of enterotoxigenic and antimicrobial-resistant strains of S. aureus in cheese constitutes a potential risk to public health. This result calls for better control of cheese contamination sources, and taking hygienic measures is necessary for food safety. More attention should be paid to animal welfare and hygiene practices in some dairy farms during manufacturing to enhance the microbiological quality of traditional cheese products.

Guava fibre characterization and effects on digestibility, fermentation products, gastrointestinal transit time and palatability of dry diets for dogs.

The use of fruit fibres as a way of reusing industrial waste is attractive and necessary, increasing the efficiency and reducing the environmental impact of the industry. This study characterized guava fibre as a fibrous ingredient and evaluated its effects when included in extruded diets for dogs on nutrient digestibility, faecal traits, fermentation products, gastrointestinal transit time and palatability. Four diets were formulated: CO (control diet, no fibrous ingredients added), GF3 (3% guava fibre), GF6 (6% guava fibre) and GF12 (12% guava fibre). The guava fibre was added to diets in replacement of maize, with small adjustments in the other ingredients to ensure similar contents of protein and fat. The diets were fed to 24 adult Beagle dogs (6 per diet) during 15 days for adaptation followed by 15 days for assessment of digestibility, fermentation end products and gastrointestinal transit time. The palatability of G6 and G12 treatments was evaluated against the CO by the two-pan test. Results were compared by analysis of variance and polynomial contrasts according to the guava fibre inclusion level (p < 0.05). The addition of guava fibre did not change nutrient intake except for the increase on dietary fibre (p < 0.001). Fibre inclusion resulted in a quadratic reduction in total tract apparent digestibility for dry matter, organic matter, crude protein and crude energy (p < 0.001), and on the metabolizable energy content of the foods (p < 0.001). Guava fibre addition did not change the faecal concentration of ammonia, lactic acid, faecal pH and branched chain fatty acids but it decreased acetic and propionic acids concentration (p < 0.01). The inclusion of 12% guava fibre did result in a faster transit time (p = 0.046) compared to the control diet. Guava fibre can be characterized as a novel insoluble non-fermentable fibre source that could be safely used in levels up to 12% in canine extruded diets.

Chemometrics of the Composition and Antioxidant Capacity of Hyptis crenata Essential Oils from Brazil.

Hyptis crenata (Pohl) ex Benth is used in traditional medicine as an analgesic to treat general pain. Six Hyptis crenata samples (Hc-1 to Hc-6) were collected in Pará state, Brazil. The leaf essential oils were obtained by hydrodistillation, and GC-MS and GC-FID were used to analyze their chemical compositions. The antioxidant capacity was measured in vitro using DPPH and carotene/linoleic acid assays. Chemometrics analysis (PCA, HCA, and clustered heat map) were used to identify the sample relationships between those collected in this study and those from the literature (Hc-7 to Hc-16) samples. According to the main chemical constituents identified in the samples described in this work and the literature, the sixteen samples were classified into ten groups. Group I was characterized by 1,8-cineole (31.0%), α-pinene (13.6%), (E)-caryophyllene (7.8%), and ß-pinene (7.6%); and Group IV was characterized by 1,8-cineole (17.4-23.5%), α-pinene (15.7-23.5%), ß-pinene (10.5-13.4%), and limonene (8.5-9.7%). Both groups are described for the first time. The total antioxidant capacity was expressed in Trolox Equivalent Antioxidant Capacity values (TEAC): TEAC of Hc-5 (551.9 mg.TE/g) and Hc-6 (475.1 mg.TE/g). In the ß-carotene/linoleic acid assay, the highest inhibition was from Hc-2 (40.0%), Hc-6 (39.0%), and Hc-3 (29.4%).

Exploring the complex role of chemokines and chemoattractants in vivo on leukocyte dynamics.

Chemotaxis is fundamental for leukocyte migration in immunity and inflammation and contributes to the pathogenesis of many human diseases. Although chemokines and various other chemoattractants were initially appreciated as important mediators of acute inflammation, in the past years they have emerged as critical mediators of cell migration during immune surveillance, organ development, and cancer progression. Such advances in our knowledge in chemokine biology have paved the way for the development of specific pharmacological targets with great therapeutic potential. Chemoattractants may belong to different classes, including a complex chemokine system of approximately 50 endogenous molecules that bind to G protein-coupled receptors, which are expressed by a wide variety of cell types. Also, an unknown number of other chemoattractants may be generated by pathogens and damaged/dead cells. Therefore, blocking chemotaxis without causing side effects is an extremely challenging task. In this review, we focus on recent advances in understanding how the chemokine system orchestrates immune cell migration and positioning at the whole organ level in homeostasis, inflammation, and infection.

Contribution of the vectorcardiogram in the differential diagnosis of Brugada electrocardiographic pattern.

BACKGROUND: The electrocardiogram (ECG) is a powerful tool for differential diagnosis among a group of pathologies with different therapeutic approaches/prognoses, the so-called J-wave syndrome. The vectorcardiogram (VCG) can be used as a complementary method to the ECG in several dubious electrocardiographic alterations. OBJECTIVE: We carried out a VCG analysis after conceiving and measuring a novel parameter (JT-distance) that allows diagnosis of the Brugada ECG pattern. METHODS: A retrospective cohort study selected ninety-six ECGs with J-point elevation in V1/V2, ECG superior leads and VCGs, all performed on the same day. A new VCG measurement by Frank method (JT-distance) was conceived and designed in transverse and right sagittal planes by 3 lines drawn 1) at the final third of the QRS loop, comprehending the J-point; 2) at the initial portion of the T loop; 3) a parallel of the J-point line at the beginning of the T loop. JT measure was determined by the distance between parallels. A validation cohort was established in a new sample of thirty-five patients. RESULTS: JT-distance ≥1.5 mm (tranverse plane) and JT-distance >1.25 mm, in the sagittal plane, differentiated Brugada type-1 from Brugada type-2, early repolarization and others, with 95% sensitivity and 68% specificity. JT-distance <1.5 mm (transverse plane) and JT >1.25 mm (sagittal plane) had 100% sensitivity and 85% specificity for Brugada type-1 diagnosis. A validation cohort showed very similar Cohen's kappa levels (0.65 and 0.77, test and validation cohorts, respectively), with overlapping 95% confidence intervals. CONCLUSIONS: The novel vectorcardiogram measurement (JT-distance) presented a new diagnostic criterion to identify Brugada pattern. Nevertheless, prospective studies should be performed by other centers to confirm these findings.

Phenotypic divergence among half-sib progenies of African mahogany.

Divergence studies are essential for the conservation of genetic resources and guidance in genetic improvement programs. This study aimed to assess the phenotypic dispersion of half-sib progenies using Gower's algorithm and the non-metric multidimensional scaling analysis based on qualitative and quantitative traits. Twenty-two half-sib progenies of African mahogany (Khaya senegalensis) were evaluated in a randomized block design with 22 treatments (progenies/origin), with 30 replications and one plant per plot, totaling 660 plants. The total tree height (cm), diameter at soil level (cm), chlorophyll A and B, presence of bifurcation, presence of lateral branches, stem tortuosity, and the phytosanitary and nutritional status according to a rating scale were evaluated. Data analysis was performed at the progeny mean level for the quantitative traits and the modal value was used for the qualitative traits. The technique used presented the distance between the progenies of K. senegalensis in a graphical form. The dispersion of the progenies in the graph is indicative that there is phenotypic variation in the progeny test, allowing to select individuals with characteristics of interest to improve mahogany for Piauí. Gower's algorithm applied to mixed data and non-metric multidimensional scaling are efficient methods to describe phenotypic similarities between progenies.

Characterization and effectiveness of a violet LED light for in-office whitening.

OBJECTIVES: This study characterized a violet LED light (V-LED; bright max whitening) tooth whitening device and evaluated its efficacy on stained enamel compared to hydrogen peroxide (HP). MATERIALS AND METHODS: Characterization of the V-LED beam profile was performed using a laser beam-profiler. The irradiance was measured throughout an exposure cycle at 0- and 8-mm distances using an integrating sphere and a spectral radiometer. Bovine enamel/dentin blocks stained with black tea (BT), cigarette smoke (CS), or without staining (CONT) were subjected to V-LED or 40% HP (n = 10/group). Color parameters (ΔL, Δa, Δb, and ΔE00) were measured using a digital spectrophotometer. Light transmission was estimated through 1-mm-thick bovine enamel slices (n = 5). ΔL, Δb, ΔE00, and irradiance were analyzed by two-way ANOVAs and Tukey's tests, Δa by Kruskal-Wallis and Mann-Whitney tests, and light transmission by t-test (α = 5%). RESULTS: Heterogeneous beam distribution was observed for the emitting V-LED chips. After 20 sequential exposures, irradiance levels were reduced 25-50%, regardless of the distance from V-LED. Localized irradiance values were statistically different between beam locations and different distances from the target. V-LED produced lower ΔE00, ΔL, Δa, and Δb values than HP for CONT and BT, with no differences for CS. Light transmittance decreased approximately 98% through 1-mm thick enamel. CONCLUSIONS: V-LED irradiance was heterogeneous and decreased throughout the exposure cycles and was also greatly reduced with increasing tip distance. V-LED produced a significantly lower whitening effect on BT and control teeth. CLINICAL RELEVANCE: This study contributes to the knowledge of V-LED and its clinical use.

Reduced Dose of Beta-Alanine Is Sufficient to Maintain Performance in Repeated Sprints.

ABSTRACT: Zandona, BA, Ramos, RA, de Oliveira, CdS, McAnulty, SR, Ferreira, LHB, Smolarek, AC, Enes, AAN, Urbinati, KMdSS, Aragon, AA, Schoenfeld, BJ, and de Souza Junior, TP. Reduced Dose of Beta-Alanine Is Sufficient to Maintain Performance in Repeated Sprints. J Strength Cond Res 36(6): 1636-1642, 2022-Beta-alanine (BA) supplementation has been shown to be effective in improving physical performance by increasing carnosine concentration. However, it is still necessary to know the effect of a maintenance dose on performance. Thus, this study aimed to investigate the effects of a maintenance dose of BA supplementation on performance. Forty-four anaerobically trained men with 23.9 ± 3.8 years of age, 176.0 ± 0.05 cm height, 81.2 ± 7.5 kg body mass, and 15.5 ± 2.9% of body fat performed a cycle ergometer test consisting of 4 sprints of 30 s with 4 minutes of active recovery. The study comprised 3 phases: (a) presupplementation, (b) supplementation with 6.4 g·d-1 BA or placebo, and (c) postsupplementation with a maintenance dose of 1.2 g·d-1 of BA or interruption of supplementation. Data were analyzed using generalized estimated equations with a priori 0.05 level of significance. The placebo group and interruption group presented a lower power (7.28 ± 0.66 and 7.71 ± 0.42 W·kg-1 vs. 8.04 ± 0.84 and 9.25 ± 1.18 W·kg-1, respectively; p < 0.05) during the third sprint in postsupplementation, whereas the maintenance group maintained the required power (7.47 ± 1.03 vs. 8.74 ± 1.07 W·kg-1; p > 0.05). The placebo group also presented higher percentage of fatigue (44.5% ± 12.3 and 44.8% ± 7.7 vs. 37.6 ± 7.2%; p = 0.021) and higher subjective perception of exertion (8.92 ± 0.90 vs. 8.00 ± 1.60; p = 0.028). Therefore, the maintenance dose of 1.2 g·d-1 BA was effective in maintaining performance, whereas a reduction in performance was observed after supplementation interruption.

Corporoplasty: A simplified technique for clitoroplasty.

INTRODUCTION: Clitoroplasty constitutes an important step in feminizing surgery for congenital adrenal hyperplasia (CAH) (1). In this video we present a technique that aims to preserve clitoral sensitivity and engorgement while minimizing the risk of neurovascular lesion. MATERIALS AND METHODS: We present a video of a three-year-old girl with history of CAH classical form, PRADER-III, who underwent clitoroplasty. After an initial endoscopic evaluation of the urogenital sinus, the clitoris was degloved and a rectangular incision was made on the ventral corpora cavernosa 15mm above the corpora bifurcation and 0.5 mm below the coronal sulcus. The cavernous tissue was partially resected. The upper and lower borders of the rectangular gap were closed by a 5-0 PDS running suture similar to the Mikulicz technique. Next, the edge of the glans was deepithelialized to reduce its size. For improved clitoral positioning, the clitoris was sutured to the pubic fat. From that point onward the procedure followed that of a standard vaginoplasty using the en-bloc technique (2-4). Thus far we have performed this technique in 33 patients, with 31 of them being girls with CAH and 2 being women with clitoral hypertrophy. CONCLUSION: Corporoplasty is a simplified technique for clitoroplasty, with the advantage being that is faster and safer than the technique that involves the dissection of the neurovascular bundle. In addition, corporoplasty has the possible benefit of preserving the cavernosal blood flow that permits the engorgement of the clitoris during sexual arousal.

Intratubular Sealer Penetration: Scanning Electron Microscopy Associated with Cathodoluminescence Analysis.

AIM: The aim of the study was to compare the ability of three endodontic sealers, Endofill (END), AH Plus (AHP), and Sealer Plus BC (SPB), to penetrate dentinal tubules. MATERIALS AND METHODS: Forty-five human teeth, single-rooted and previously instrumented mandibular premolars, were randomly divided into three experimental groups (n = 15): END (n = 15), AHP (n = 15), and SPB (n = 15). After obturation, dental sections were performed horizontally, at 2 and 5 mm from the root apex. The samples were analyzed by scanning electron microscopy associated with cathodoluminescence. Percentage penetration (PP%) and maximum penetration depth (MPD) of the sealers were evaluated by the Kruskal-Wallis and Mann-Whitney tests, for general and paired data, respectively. The Wilcoxon test was applied to analyze the differences between the 5 and 2 mm distances. A 5% significance level was adopted. RESULTS: As for PP%, AHP and SPB were similar (p = 0.127) and presented higher values than END (AHP, p = 0.024 and SPB, p <0.001); with regard to MPD, AHP and SPB did not differ either (p = 0.450), but were higher than END (p <0.001); in both analyses, penetration was greater at 5 mm than at 2 mm (p <0.001). CONCLUSION: SPB showed satisfactory performance in penetrating dentinal tubules, being similar to AHP, and superior to END. CLINICAL SIGNIFICANCE: Greater penetration of sealer into the dentinal tubules may increase the chance of successful endodontic treatment.

Use of antioxidants to restore bond strength after tooth bleaching with peroxides.

This review compiles the literature on the antioxidants used after tooth bleaching with either low or high-concentrated carbamide and hydrogen peroxide to recover the bond strength. Antioxidants used in bleached teeth are mainly natural and non-enzymatic, except for catalase. Commonly, antioxidants are applied to remove any reactive oxygen species (ROS) residues left from bleaching gels, which adversely affect adhesive procedures, such as restorations or orthodontic brackets bonding. Even though sodium ascorbate, the most thoroughly investigated antioxidant, showed the most efficient bond strength recovery at 10% concentration, its performance depends on the type of solution and the application time. Natural extracts, such as proanthocyanidins and green tea, showed satisfactory results in the reversal of bond strength at 5% and 10% concentrations, respectively. Sodium ascorbyl phosphate, α-tocopherol, and catalase exhibited promising results, but further research is required. The adhesive system type plays an important role in the outcome of enamel bond strength after the antioxidant application. The postponement of either restorations or orthodontic brackets cementation following bleaching procedures seems to be efficiently replaced by antioxidant application prior to bonding procedures. However, the efficacy of using an antioxidant to recover bond strength depends on its type and application time.

Oral shedding of CMV and HSV-1 in hematopoietic stem cell transplantation patients.

OBJECTIVES: To evaluate the oral shedding of herpesviruses in patients undergoing hematopoietic stem cell transplantation (HSCT) and correlate it with oral mucositis (OM). METHODS: Saliva samples were collected before the HSCT and on day D + 8. Multiplex Polymerse Chain Reaction (PCR) was performed to detect herpes simplex virus (HSV)-1 and HSV-2, Epstein-Barr virus (EBV), Cytomegalovirus (CMV), Variella-zoster virus (VZV), and human herpesvirus (HHV)-6, HHV-7, and HHV-8. OM was assessed according to WHO criteria. RESULTS: Thirty one patients were enrolled, in which 20 of 31 (64.5%) were males; median age was 50 (21-70) years; 16 of 31 (51.6%) underwent allo-HSCT; and 15 of 31 (48.4%) underwent auto-HSCT. On D + 8, OM grades III and IV were observed in 8 of 31 (25.8%) patients. In the first salivary collection, EBV was found in 24 of 31 (77.4%), followed by HHV-6 (7/31, 22.6%) and HHV-7 (8/31 25.8%). In the second collection, EBV was found in 24 of 27(89%), followed by HSV-1 (8/27, 30%) and CMV, HHV-6, and HHV-7 (5/27, 18.5%, each one). On D + 8, OM grades II and IV were associated with the presence of HSV-1. HSV-1 was also associated with worsening degrees of OM on D + 15. CONCLUSION: The presence of HSV-1 and CMV in oral samples was more frequent on day D + 8 after HSCT. HSV-1 detection was associated with severity and worsening of OM. HSV-1 and CMV seem to be associated with oral dysbiosis due to HSCT.

High throughput screening of technological and biopreservation traits of a large set of wild lactic acid bacteria from Brazilian artisanal cheeses.

This study aimed to evaluate technological (acidification, proteolysis, lipolysis, resistance to low pH, NaCl, and bile salts) and biopreservation (antimicrobial activity against foodborne pathogens) features of 1002 LAB by high throughput screening (HTS) methods. The LAB was isolated from 11 types of Brazilian artisanal cheeses (BAC) marketed in the main 5 producing regions. Remarkable intra-species variability in acidification rates have been found, which was most pronounced between isolates from Mina's artisanal cheeses, Caipira and Coalho cheeses. Lacticaseibacillus paracasei and Levilactobacillus brevis showed the fastest acidification rate; however, all isolates showed slower acidification rates than a lactococcal control strain (4.3 × lower). When testing inhibitory effects, > 75% of LAB isolates could inhibit the growth of Staphylococcus aureus ATCC 19095 and Listeria monocytogenes ATCC 7644. Two of these isolates, identified as Lactiplantibacillus plantarum and Lentilactobacillus buchneri, the sterile and neutral supernatants alone, were sufficient to inhibit L. monocytogenes growth. Principal component analysis (PCA) allowed the identification of functional groups based on proteolytic and lipolytic activity, osmotic stress resistance, and inhibition of L. monocytogenes. The type of cheese the isolates were recovered from influenced properties such as anti-listerial compounds and lipolytic enzyme production. The use of HTS and multivariate statistics allowed insights into a diverse set of LAB technological and biopreservation properties. These findings allow a profound knowledge of the heterogeneity of a large set of isolates, which can be further used to design starter cultures with varied and combined properties, such as biopreservation and technological features. Besides that, HTS makes it possible to analyze a vast panel of LAB strains, reducing costs and time within laboratory analysis, while avoiding the loss of information once all LAB are tested at the same time (differently from the traditional labor-intensive approach, in which a few numbers of strains is tested per time).

Technological Advancements in Monoclonal Antibodies.

Biopharmaceuticals are innovative solutions that have revolutionized the treatment of important chronic diseases and malignancies. The approval of biosimilar products has become a complex and balanced process, and there are versions of drugs with established biosimilarity that can offer a more accessible treatment option to patients. The objective of this work was to identify the advancement of these technologies by means of patent and article analysis based on technological and scientific prospection. In patent document recovery, Derwent Innovation Index (DWPI) and PatentInspiration databases were used. The research was based on the search of the selected terms in the title, summary, and claims of the documents through a search strategy containing IPC code and keywords. In articles recovery, the Web of Science tool was used in the search of scientific publications dated from the last 5 years. The search resulted in a total of 2295 individual patent documents and 467 families using DWPI database, 769 individual patents and 205 families using PatentInspiration, and 2602 articles using Web of Science database. Additionally, this work describes the number of organizations that contribute to this area, where they are, how much development they have undergone, and the inventors/authors involved. Based on the number of publications registered, there is an important prominence for scientific research in mAbs. In terms of innovation, it is expected that several therapeutic drugs are already under regulatory review, which will allow drugs to be approved over the next few years and will thus generate a continuous flow of new products based on immunotherapies, mAbs, and biosimilar drugs. These drugs have become essential weapons for the treatment of significant diseases, and the increasing trend in the number of related scientific and technological publications contributes to making these therapies available to the greatest number of people.

A Simple Method for Evaluating the Bioactive Phenolic Compounds' Presence in Brazilian Craft Beers.

There are a significant number of analytical methodologies employing different techniques to determine phenolic compounds in beverages. However, these methods employ long sample preparation processes and great time consumption. The aim of this paper was the development of a simple method for evaluating the phenolic compounds' presence in Brazilian craft beers without a previous extraction step. Catechin, caffeic acid, epicatechin, p-coumaric acid, hydrated rutin, trans-ferulic acid, quercetin, kaempferol, and formononetin were analyzed in fifteen different craft beers. The method showed good linearity (R2 ≥ 0.9966). The limit of detection ranged from 0.08 to 0.83 mg L-1, and limits of quantification were between 0.27 and 2.78 mg L-1. The method showed a satisfactory precision (RSD ≤ 16.2%). A good accuracy was obtained by the proposed method for all phenolic compounds in craft beer (68.6% ˂ accuracy ˂ 112%). Catechin showed higher concentrations (up to 124.8 mg L-1) in the samples, followed by epicatechin (up to 51.1 mg L-1) and caffeic acid (up to 8.13 mg L-1). Rutin and formononetin were observed in all analyzed samples (0.52 mg L-1 to 2.40 mg L-1), and kaempferol was less present in the samples. The presence of plant origin products was determinant for the occurrence of the highest concentrations of phenolic compounds in Brazilian craft beers.

Effects of crude protein and sodium intake on water turnover in cats fed extruded diets.

The comprehension of strategies to increase urine production may be important, especially in kibble diets to prevent urolithiasis in cats. The effects of increasing amounts of crude protein (CP) and sodium on the water turnover of cats were evaluated using the water balance (WB) method and the deuterium dilution technique. The study followed a randomized block design, with three blocks of eight cats, two cats per food type in each block, and six cats per food. Four extruded diets with different amounts of CP and sodium were evaluated (on DM basis): 28% CP and 0.58% sodium; 39% CP and 0.64% sodium; 52% CP and 0.76% sodium; and 64% CP and 0.87% sodium. Cats were individually housed in cages for 8 days to measure WB, urea excretion, and faecal and urine characteristics. Deuterium oxide was used to evaluate water turnover, and during the period cats were housed in a collective cattery. The data were analysed by an F test, and the means were compared by polynomial contrasts. The ɑ level of significance was set at 0.05. The methods were compared by Pearson correlation, and Bland and Altman analysis. The increase in the CP content elevated linearly the renal excretion of urea (p < .001), and, together with the higher sodium intake, elevated the renal solute load, which resulted in a linear increase in urine production and water intake (p < .01). The urine density, metabolic water, and faecal and insensible water losses did not differ (p > .05). The water flux increased linearly when using the deuterium method (p < .001), but the obtained values were 20.85 ± 11.11 ml/cat/day higher than those verified using the WB method (p = .001). Higher CP and sodium amounts in dry diets increased the urine production and water consumption of cats, and this can be explored as a possible option to increase urination.

Nonmulberry Silk Based Ink for Fabricating Mechanically Robust Cardiac Patches and Endothelialized Myocardium-on-a-Chip Application.

Bioprinting holds great promise towards engineering functional cardiac tissue constructs for regenerative medicine and as drug test models. However, it is highly limited by the choice of inks that require maintaining a balance between the structure and functional properties associated with the cardiac tissue. In this regard, we have developed a novel and mechanically robust biomaterial-ink based on non-mulberry silk fibroin protein. The silk-based ink demonstrated suitable mechanical properties required in terms of elasticity and stiffness (~40 kPa) for developing clinically relevant cardiac tissue constructs. The ink allowed the fabrication of stable anisotropic scaffolds using a dual crosslinking method, which were able to support formation of aligned sarcomeres, high expression of gap junction proteins as connexin-43, and maintain synchronously beating of cardiomyocytes. The printed constructs were found to be non-immunogenic in vitro and in vivo. Furthermore, delving into an innovative method for fabricating a vascularized myocardial tissue-on-a-chip, the silk-based ink was used as supporting hydrogel for encapsulating human induced pluripotent stem cell derived cardiac spheroids (hiPSC-CSs) and creating perfusable vascularized channels via an embedded bioprinting technique. We confirmed the ability of silk-based supporting hydrogel towards maturation and viability of hiPSC-CSs and endothelial cells, and for applications in evaluating drug toxicity.

Validation of saliva and urine use and sampling time on the doubly labelled water method to measure energy expenditure, body composition and water turnover in male and female cats.

Less invasive protocols are necessary to study energy expenditure (EE) of cats living in homes for expressing their normal living conditions. The present study compared sampling times and the use of saliva, urine and blood to measure 2H and 18O to apply the doubly labelled water method. In the first study, four cats were used to evaluate the enrichment (2, 4, 6, 7 and 8 h) and elimination (2, 4, 6, 8, 10, 12, 14, 16, 18 and 20 d) of 2H and 18O (subcutaneously injected). The maximum enrichment was after 5 h (R2 0·82) of injection, with an Ln linear elimination of both isotopes (P < 0·001; R2 0·99). The results of EE were similar, regardless of the sampling time used (P = 0·999). In the second study, seven male cats and seven female cats were used. Before and after isotope injection (5 h, 7 d, 10 d and 14 d), blood, saliva and urine were collected. Isotope enrichment was lower in urine (P < 0·05) and at the similar level in blood and saliva. Isotope elimination was similar for all fluids (P < 0·473). The EE calculated with blood and saliva was similar but higher for urine (P = 0·015). According to Bland-Altman statistics, blood and saliva presented low bias and high correlation (P < 0·001), but this was not observed for urine (P = 0·096). Higher EE was observed for male cats (384 (se 39) kJ/kg0·67 per d) than for female cats (337 (se 34) kJ/kg0·67 per d; P < 0·05). The sampling time for the method is flexible, and saliva can be used as a substitute for blood.