Roasted yerba mate (Ilex paraguariensis) infusions in bovine milk model before and after in vitro digestion: Bioaccessibility of phenolic compounds, antioxidant activity, protein-polyphenol interactions and bioactive peptides.

Yerba mate is increasingly acknowledged for its bioactive properties and is currently being incorporated into various food and pharmaceutical products. When roasted, yerba mate transforms into mate tea, consumed as a hot aqueous infusion, and has gained popularity. This study investigated the bioaccessibility of phenolic compounds, protein-polyphenol interactions, antioxidant activity, and bioactive peptides in roasted yerba mate infusions, utilizing whole, semi-skimmed, and skimmed bovine milk models. The phytochemical profile of roasted yerba mate was analyzed in infusions with water and milk (whole, semi-skimmed, and skimmed), before and after in vitro digestion, identifying 18 compounds that exhibited variations in composition and presence among the samples. Bioavailability varied across different milk matrices, with milk being four times more efficient as a solvent for extraction. Gastric digestion significantly impacted (p < 0.05) the release of phenolic compounds, such as chlorogenic acid and rutin, with only chlorogenic acid remaining 100 % bioavailable in the infusion prepared with skimmed milk. Protein-polyphenol interaction did not influence protein digestion in different infusions, as there was a similarity in the hydrolysis pattern during the digestive process. Changes in antioxidant activity during digestion phases, especially after intestinal digestion in milk infusions, were related to alterations in protein structures and digestive interactions. The evaluation of total phenolic compounds highlighted that skimmed milk infusion notably preserved these compounds during digestion. Peptidomic analysis identified 253, 221, and 191 potentially bioactive peptides for whole, semi-skimmed, and skimmed milk-digested infusions, respectively, with a focus on anti-inflammatory and anticancer activities, presenting a synergistic approach to promote health benefits. The selection of milk type is crucial for comprehending the effects of digestion and interactions in bioactive compound-rich foods, highlighting the advantages of consuming plant infusions prepared with milk.

Validation and optimization of the loop-mediated isothermal amplification (LAMP) technique for rapid detection of wheat stripe mosaic virus, a wheat-infecting pathogen.

BACKGROUND: Wheat stripe mosaic virus (WhSMV) is a significant wheat pathogen that causes substantial yield losses in Brazil and other countries. Although several detection methods are available, reliable and efficient tools for on-site WhSMV detection are currently lacking. In this study, a Loop-Mediated Isothermal Amplification (LAMP) method was developed for rapid and reliable field detection of WhSMV. We designed WhSMV-specific primers for the LAMP assay and optimized reaction conditions for increased sensitivity and specificity using infected plant samples. RESULTS: We have developed a diagnostic method utilizing the Loop-Mediated Isothermal Amplification (LAMP) technique capable of rapidly and reliably detecting WhSMV. The LAMP assay has been optimized to enhance sensitivity, specificity, and cost-effectiveness. CONCLUSION: The LAMP assay described here represents a valuable tool for early WhSMV detection, serving to mitigate the adverse economic and social impacts of this viral pathogen. By enabling swift and accurate identification, this assay can significantly improve the sustainability of cereal production systems, safeguarding crop yields against the detrimental effects of WhSMV.

Caloric Restriction Rejuvenates Skeletal Muscle Growth in Heart Failure With Preserved Ejection Fraction.

Heart failure with preserved ejection fraction (HFpEF) is a major clinical problem, with limited treatments. HFpEF is characterized by a distinct, but poorly understood, skeletal muscle pathology, which could offer an alternative therapeutic target. In a rat model, we identified impaired myonuclear accretion as a mechanism for low myofiber growth in HFpEF following resistance exercise. Acute caloric restriction rescued skeletal muscle pathology in HFpEF, whereas cardiac therapies had no effect. Mechanisms regulating myonuclear accretion were dysregulated in patients with HFpEF. Overall, these findings may have widespread implications in HFpEF, indicating combined dietary with exercise interventions as a beneficial approach to overcome skeletal muscle pathology.

Cactus species and tree forage hay in the diet of dairy goats.

The purpose of this study was to evaluate the influence of the association of Opuntia stricta (OS) or Nopalea cochenillifera (NC) with Moringa hay (MY) or Leucaena hay (LY) on nutrient intake, ingestive behavior, serum biochemistry, and milk production and composition of dairy goat. Eight multiparous Saanen goats were used, with an average of 68 ± 8 days in milk and 48.5 ± 6 kg of initial weight, fed with the following treatment diets: OS + MY; OS + LY; NC + MY, and NC + LY. The treatments were distributed in a double Latin square, in a 2 × 2 factorial scheme (two varieties of cactus and two of hay). Goats fed with NC had higher (P < 0.05) intakes of dry matter, crude protein, neutral detergent fiber (NDF), and non-fibrous carbohydrates. There was no influence (P > 0.05) on milk production, but goats fed with OS produced milk with higher (P < 0.05) fat content and presented higher (P < 0.05) feed efficiency. There was no effect (P > 0.05) of the hay type on the nutrient intake and milk production and composition of goats, but the animals fed with MY had higher (P < 0.05) serum cholesterol levels and greater efficiency of NDF rumination. Goats fed with OS showed greater feeding and rumination efficiency. Our results indicated that both cactus species could be successfully included in the diet of low yielding dairy goats fed 18.4% concentrate.

Oral habits, sociopsychological orthodontic needs, and sociodemographic factors perceived by caregivers impact oral health-related quality of life in children with and without autism?

BACKGROUND: Caregivers play a crucial role in assessing the oral health-related quality of life (OHRQoL) of young individuals with autism spectrum disorder (ASD). AIM: This study assessed the impact of sociodemographic and oral conditions on OHRQoL and family dynamics in young individuals with and without autism, as perceived by guardians. DESIGN: This comparative cross-sectional study included young individuals aged 6 to 14 years and their guardians. Data were collected at a specialized institution and dental schools. Guardians completed the Parental-Caregiver Perceptions Questionnaire (P-CPQ), Family Impact Scale (FIS), and sociodemographic and oral habits questionnaires. The sociopsychological need for orthodontic treatment was assessed using the Index of Orthodontic Treatment Need (IOTN). RESULTS: The sample included 144 youths and caregivers. The ASD group had higher P-CPQ and FIS scores. Factors associated with poorer perceived OHRQoL included higher youth age, lower caregiver education, higher IOTN scores, teeth clenching (RR = 1.20; 95% CI: 1.01-1.41), and lip sucking. Lower parental education (RR = 1.75; 95% CI: 1.10-2.80) and higher IOTN scores from the caregiver's perspective impacted family dynamics. CONCLUSION: Caregivers of young individuals with ASD perceived a lower OHRQoL, and families in this group were more affected by sociodemographic and oral conditions.

Antiaging effects of a skin care formulation containing nanoencapsulated antioxidants: A clinical, in vitro, and ex vivo study.

BACKGROUND: The development of effective cosmetic products for the reduction of the signs of skin aging is a complex process which requires an optimized combination of ingredients and specialized systems to deliver the actives to the skin layers. AIM: To evaluate the tolerance and antiaging clinical efficacy of a cosmetic formulation containing a blend of nanoencapsulated antioxidants: ascorbyl palmitate, resveratrol, tocopherol, caffeine, carnosine, and niacinamide. METHODS: Clinical efficacy was determined by subjective and instrumental analyses of collagen synthesis by fluorescence spectroscopy, by three-dimensional imaging analysis of suborbital edema, and by analysis of skin hydration and sebum content by biophysical techniques-Corneometer® and Sebumeter®. RESULTS: The studied formulation was safe and effective for the improvement of skin appearance by increasing collagen synthesis and skin moisturizing and by reducing facial blemishes, swelling, and oiliness. A preclinical exploratory approach using an experimental model of human cell and skin cultures agreed with the observed antiaging effects, identifying mechanisms related to the containment of oxidative stress, reduction of melanin production, increased synthesis of type I procollagen, and regulation of the epidermal cohesion protein filaggrin. CONCLUSIONS: The skin benefits obtained resulted from the combination of the ingredients in the formulation and the nanoencapsulation-based delivery system, which favors the solubility, safety, efficacy, and bioavailability of the preparation to the skin.

Natural variation in a short region of the Acidovorax citrulli type III-secreted effector AopW1 is associated with differences in cytotoxicity and host adaptation.

Bacterial fruit blotch, caused by Acidovorax citrulli, is a serious disease of melon and watermelon. The strains of the pathogen belong to two major genetic groups: group I strains are strongly associated with melon, while group II strains are more aggressive on watermelon. A. citrulli secretes many protein effectors to the host cell via the type III secretion system. Here we characterized AopW1, an effector that shares similarity to the actin cytoskeleton-disrupting effector HopW1 of Pseudomonas syringae and with effectors from other plant-pathogenic bacterial species. AopW1 has a highly variable region (HVR) within amino acid positions 147 to 192, showing 14 amino acid differences between group I and II variants. We show that group I AopW1 is more toxic to yeast and Nicotiana benthamiana cells than group II AopW1, having stronger actin filament disruption activity, and increased ability to induce cell death and reduce callose deposition. We further demonstrated the importance of some amino acid positions within the HVR for AopW1 cytotoxicity. Cellular analyses revealed that AopW1 also localizes to the endoplasmic reticulum, chloroplasts, and plant endosomes. We also show that overexpression of the endosome-associated protein EHD1 attenuates AopW1-induced cell death and increases defense responses. Finally, we show that sequence variation in AopW1 plays a significant role in the adaptation of group I and II strains to their preferred hosts, melon and watermelon, respectively. This study provides new insights into the HopW1 family of bacterial effectors and provides first evidence on the involvement of EHD1 in response to biotic stress.

Simplified assessment of castration-induced pain in pigs using lower complexity algorithms.

Pigs are raised on a global scale for commercial or research purposes and often experience pain as a by product of management practices and procedures performed. Therefore, ensuring pain can be effectively identified and monitored in these settings is critical to ensure appropriate pig welfare. The Unesp-Botucatu Pig Composite Acute Pain Scale (UPAPS) was validated to diagnose pain in pre-weaned and weaned pigs using a combination of six behavioral items. To date, statistical weighting of supervised and unsupervised algorithms was not compared in ranking pain-altered behaviors in swine has not been performed. Therefore, the aim of this study was to verify if supervised and unsupervised algorithms with different levels of complexity can improve UPAPS pain diagnosis in pigs undergoing castration. The predictive capacity of the algorithms was evaluated by the area under the curve (AUC). Lower complexity algorithms containing fewer pain-altered behaviors had similar AUC (90.1-90.6) than algorithms containing five (89.18-91.24) and UPAPS (90.58). In conclusion, utilizing a short version of the UPAPS did not influence the predictive capacity of the scale, and therefore it may be easier to apply and be implemented consistently to monitor pain in commercial and experimental settings.

Biofertilizers from wastewater treatment as a potential source of mineral nutrients for growth of amaranth plants.

Exploring alternative fertilizers is crucial in agriculture due to the cost and environmental impact of inorganic options. This study investigated the potential of sewage-derived biofertilizers on the growth and physiology of Amaranthus cruentus plants. Various treatments were compared, including control treatments with inorganic fertilizer and treatments with biofertilizers composed of microalgae, biosolids and reclaimed water. The following traits were investigated: photosynthetic pigments, gas exchange, growth, and leaf nutrient concentrations. The results showed that the concentrations of N, P, Cu, Fe Zn and Na nutrients, in the dry microalgae and biosolids, were quite high for the needs of the plants. The wet microalgae presented high concentration of Cu, Fe and Zn nutrients while reclaimed water contained high concentration of N, K, Ca and S. Na and Zn nutrients increased in the leaf of plants treated with dry microalgae and biosolid, respectively. At the beginning of the flowering phase, total chlorophyll and carotenoids contents were lower for plants grown with wet microalgae while for plants grown with higher doses of biosolid or reclaimed water total chlorophyll was increased, and carotenoids were not affected. Lower photosynthetic pigments under wet microalgae resulted in lower photosynthetic rates. On the other hand, amendments with dry microalgae and biosolid increased photosynthetic rates with the biosolid being the most effective. Higher applications of biosolid, wet and dry microalgae produced a considerable increase in shoot biomass of amaranth, with the dry microalgae being the most effective. Additionally, reclaimed water obtained after tertiary treatment of sewage with microalgae and biosolids applied alone showed promising effects on plant growth. Overall, these findings suggest that organic fertilizers derived from sewage treatment have the potential to enhance plant growth and contribute to sustainable agricultural practices.

Comparative cross-sectional study of socio-psychological orthodontic needs and oral habits in Autism Spectrum Disorder.

OBJECTIVE: This cross-sectional study analyzed the socio-psychological need for orthodontic treatment in children and adolescents with and without Autism Spectrum Disorder (ASD) and its association with deleterious oral habits. METHODS: Children/adolescents aged 6-14 years old, with and without ASD, and their respective caregivers were included. Caregivers completed a questionnaire addressing children and adolescents' deleterious oral habits. To assess the socio-psychological need for orthodontic treatment, we utilized the aesthetic component of the Index of Orthodontic Treatment Needs (IOTN). Caregivers and an orthodontist independently used the standardized IOTN aesthetic attractiveness scale containing ten intraoral photographs and compared them with the most similar condition of the child/adolescent. Data collection occurred at two universities in northeastern Brazil and at an ASD referral center. Statistical analyses encompassed descriptive statistics, Shapiro-Wilk, Kruskal-Wallis, Mann-Whitney, and Chi-squared tests with linear trend (α5%). RESULTS: The study involved 144 participants, evenly divided into two groups (ASD and non-ASN). The ASD group showed higher average IOTN-AC scores as evaluated by both professionals (mean score = 4.78 [± 2.34], p = .182) and caregivers (mean score = 4.31 [± 2.71], p = .992). Caregivers reported notably elevated IOTN-AC scores in cases where onychophagy was absent (p = .049). CONCLUSIONS: Patients with ASD are indicated as having a relatively higher socio-psychological need for orthodontic treatment.

Bridging the Gap: Exploring the Preclinical Potential of Pereskia grandifolia in Metabolic-Associated Fatty Liver Disease.

Metabolic-associated fatty liver disease (MAFLD) is a complex condition characterized by steatosis and metabolic disturbances. Risk factors such as diabetes, cigarette smoking, and dyslipidaemia contribute to its development and progression. Effective and safe therapies for MAFLD are urgently needed. Pereskia grandifolia has shown potential as an alternative treatment, but its effectiveness against liver disease remains unexplored. This research aims to determine the hepatoprotective properties of P. grandifolia using a model of MAFLD. The study was carried out through various phases to assess the safety and efficacy of the ethanol-soluble fraction of P. grandifolia. Initially, an in vitro assay was performed to assess cell viability. This was followed by an acute toxicity test conducted in rats to determine the safety profile of the extract. Subsequently, the anti-inflammatory properties of P. grandifolia were examined in macrophages. For the MAFLD study, diabetic Wistar rats were made diabetic and exposed to a high fat diet and cigarette smoke, for 4 weeks. During the last 2 weeks, the rats were orally given either the vehicle (negative control group; C-), P. grandifolia (30, 100, and 300 mg/kg), or insulin in addition to simvastatin. A basal group of rats not exposed to these risk factors was also assessed. Blood samples were collected to measure cholesterol, triglycerides, glucose, ALT, and AST levels. Liver was assessed for lipid and oxidative markers, and liver histopathology was examined. P. grandifolia showed no signs of toxicity. It demonstrated anti-inflammatory effects by inhibiting phagocytosis and macrophage spreading. The MAFLD model induced liver abnormalities, including increased AST, ALT, disrupted lipid profile, oxidative stress, and significant hepatic damage. However, P. grandifolia effectively reversed these changes, highlighting its potential as a therapeutic agent. These findings emphasize the significance of P. grandifolia in mitigating hepatic consequences associated with various risk factors.

Association between the number of altered late potential criteria and increased arrhythmic risk in Brugada syndrome patients.

BACKGROUND: Brugada syndrome (BrS) is associated with abnormal electrophysiological properties at right ventricular epicardium, consisting of fragmented electrograms extending well beyond QRS termination. We aimed to evaluate the utility of signal-averaged electrocardiogram (SA-ECG) for the noninvasive assessment of late potentials (LP) and risk stratification of BrS patients. METHODS: A prospective, observational, single-center study of BrS patients is submitted to SA-ECG with the determination of the total filtered QRS duration (fQRS), root mean square voltage of the 40 ms terminal portion of the QRS (RMS40), and duration of the low-amplitude electric potential component of the terminal portion of the QRS (LAS40). LP were considered positive when above standard cut-offs: fQRS > 114 ms, RMS40 < 20 µV, and LAS40 > 38 ms. The rates of malignant arrhythmic events (MAEs), defined as sudden death or appropriate shocks, were compared in relation to clinical characteristics and SA-ECG findings. RESULTS: A total of 106 BrS patients (mean age, 48 ± 12 years, 67.9% male) were studied, 49% with type-1 spontaneous pattern and 81% asymptomatic. During a median follow up of 4.7 years, 10 patients (7.1%) suffered MAEs, including 4 sudden deaths. The presence of LP was significantly associated with the arrhythmic risk, which increased with the number of altered LP criteria. In comparison to the patients who had none or 1 altered LP criterium, MAE risk was 4.7 times higher in those with 2 altered criteria and 9.4 times higher in those with 3 altered LP criteria. CONCLUSIONS: SA-ECG may be a useful tool for risk stratification in BrS. The presence of 2 or 3 abnormal LP criteria could identify a subset of asymptomatic patients at high risk of arrhythmic events.

When it is too much: Identifying butamben excess on the surface of pharmaceutical preformulation samples by Raman mapping.

Butamben is a topical local anesthetic which formulation in lipid-based drug delivery systems (DDS) is challenging due to its affinity for hydrophilic excipients. This means that a medium polarity excipient is preferred for the development of a stable nanostructured lipid carrier (NLC) formulation. In turn, in NLC, the type and number of excipients will determine the active pharmaceutical ingredient (API) solubility and the maximum drug upload. To solve this dilemma and get the best formulation, a throughout screening study to evaluate API solubilization in different excipients was carried out. Subsequently, excipients with different solubilization capacities were selected for microscopic evaluation by Raman mapping, and in turn analysis of the distributional homogeneity index (DHI) and standard deviation of the histograms allowed solving the posed question. Design of experiments (DoE) was employed to understand better the interactions between the excipients; linear and higher-order models were obtained with R2 above of 0.8824. Even though DHI is a good parameter to be used as response, an API concentration higher than 30% (w/w) provided a homogeneous surface in case of good miscibility and, in this case, this parameter needs to be employed with an inspection and/or evaluation of other parameters. A curve of concentration vs. mean scores of images proved to be an alternative to identify the saturation/limit of linear range.

Facile Access to Structurally Diverse Antimalarial Indoles Using a One-Pot A3 Coupling and Domino Cyclization Approach.

A multistep and diversity-oriented synthetic route aiming at the A3 coupling/domino cyclization of o-ethynyl anilines, aldehydes and s-amines is described. The preparation of the corresponding precursors included a series of transformations, such as haloperoxidation and Sonogashira cross-coupling reactions, amine protection, desilylation and amine reduction. Some products of the multicomponent reaction underwent further detosylation and Suzuki coupling. The resulting library of structurally diverse compounds was evaluated against blood and liver stage malaria parasites, which revealed a promising lead with sub-micromolar activity against intra-erythrocytic forms of Plasmodium falciparum. The results from this hit-to-lead optimization are hereby reported for the first time.

Genetic diversity and molecular evolution of Plasmodium vivax Duffy Binding Protein and Merozoite Surface Protein-1 in northwestern Thailand.

The local diversity and population structure of malaria parasites vary across different regions of the world, reflecting variations in transmission intensity, host immunity, and vector species. This study aimed to use amplicon sequencing to investigate the genotypic patterns and population structure of P. vivax isolates from a highly endemic province of Thailand in recent years. Amplicon deep sequencing was performed on 70 samples for the 42-kDa region of pvmsp1 and domain II of pvdbp. Unique haplotypes were identified and a network constructed to illustrate genetic relatedness in northwestern Thailand. Based on this dataset of 70 samples collected between 2015 and 2021, 16 and 40 unique haplotypes were identified in pvdbpII and pvmsp142kDa, respectively. Nucleotide diversity was higher in pvmsp142kDa than in pvdbpII (π = 0.027 and 0.012), as was haplotype diversity (Hd = 0.962 and 0.849). pvmsp142kDa also showed a higher recombination rate and higher levels of genetic differentiation (Fst) in northwestern Thailand versus other regions (0.2761-0.4881). These data together suggested that the genetic diversity of P. vivax in northwestern Thailand at these two studied loci evolved under a balancing selection, most likely host immunity. The lower genetic diversity of pvdbpII may reflect its stronger functional constrain. In addition, despite the balancing selection, a decrease in genetic diversity was observed. Hd of pvdbpII decreased from 0.874 in 2015-2016 to 0.778 in 2018-2021; π of pvmsp142kDa decreased from 0.030 to 0.022 over the same period. Thus, the control activities must have had a strong impact on the parasite population size. The findings from this study provide an understanding of P. vivax population structure and the evolutionary force on vaccine candidates. They also established a new baseline for tracking future changes in P. vivax diversity in the most malarious area of Thailand.

Detection of skin wrinkles and quantification of roughness using a novel image processing technique from a dermatoscope device.

OBJECTIVE: Cutaneous relief analysis is crucial in the development of new skincare products, as well as in the evaluation of dermatological treatments. The analysis can be performed by qualitative or quantitative methods. We propose a new algorithm to detect wrinkles and quantify skin roughness by image processing from a dermatoscope. METHODS: A clinical study was carried out with 33 research participants, and images were collected with the dermatoscope and PRIMOS equipment for wrinkle evaluation at two different times: Day 0 (D0) and 45 days (D45) after the use of a dermocosmetic product. Later, a new algorithm was developed to detect wrinkles in the acquired images by applying filters and image transformations that generate a segmented image highlighting the wrinkles. A roughness calculation method is proposed from the pixels belonging to wrinkles. RESULTS: Correlation between the values obtained by the PRIMOS equipment and the proposed system was verified. No correlation was found for data obtained at D0; however, there was correlation at time D45 by Spearman's similarity coefficient. By comparing roughness between times D0 and D45, the treatment was statistically significant for both PRIMOS and the proposed methodology data. CONCLUSION: The wrinkle detection algorithm, in addition to the roughness calculation, demonstrated a sensitivity comparable to the PRIMOS system in evaluating the effectiveness of the dermocosmetic treatment. SIGNIFICANCE: Considering the simplicity of the dermatoscope design compared to other established devices such as PRIMOS, the proposed system is promising as an alternative for dermatological evaluations.

Rooftop water harvesting for managed aquifer recharge and flood mitigation in tropical cities: Towards a strategy of co-benefit evaluations in João Pessoa, northeast Brazil.

Intense urbanisation in many coastal areas has led to intensification of groundwater consumption, while reducing permeable areas and increasing the frequency and magnitude of flooding. Among the potential strategies to compensate for these adverse effects, which are expected to become worse as a result of climate change, rooftop rainwater harvesting (RWH) in combination with managed aquifer recharge (MAR), may be indicated. This work investigated the performance of different configurations of such a system, tested as a twofold sustainable stormwater and domestic water management tool in a tropical metropole (João Pessoa, Brazil). This area located over a sedimentary aquifer system illustrates the water security challenges of densely urbanised areas in southern cities. To that end, several configurations of rooftop catchments and storage volumes were evaluated, by simulating a MAR-RWH system connected to the regional unconfined aquifer (Barreiras Formation) through a 6″ diameter injection well. Rainfall-runoff-recharge processes and water balances were simulated using monitored high-temporal resolution rainfall data. The results showed that catchments ranging from 180 to 810 m2, connected to tanks from 0.5 to 30.0 m³, are the optimal solutions in terms of efficient rainwater retention and peak flow reduction. These solutions provided mean annual estimates of aquifer recharge between 57 and 255 m³/yr from 2004 to 2019. The results of this study highlight the opportunity for MAR schemes to reconcile stormwater management and water supply goals.

Evaluation of patients treated by telemedicine in the beginning of the COVID-19 pandemic in São Paulo, Brazil: A non-randomized clinical trial preliminary study.

We performed a pilot open-label, non-randomized controlled clinical trial in a clinic in São Paulo, Brazil in the beginning of the COVID-19 pandemic. "This medical pilot project was carried out during the pandemic of a new and unknown agent. It was necessary to find a new and safe therapeutic approach for pathogens with high potential for severity and contamination. The repositioning of safe and accessible pre-existing and approved medications and the telemedicine approach improved treated covid patients' symptoms and reduced the risk of disease transmission. The emergency application of a new medical technology was the major limitation of the study. This innovative care model is a low-cost safe strategy, and we understand that applicability can be expanded to other regions in emergency situations." The 187 patients of the study (mean age of 37.6 ± 15,6 years) were divided into four groups: (1) asymptomatic, (2) mild symptoms, (3) moderate symptoms and (4) severe symptoms and were followed up for five days. A drug intervention was performed in group 3 and the patients of Group 4 were oriented to seek hospital care. Of all the patients, 23.0% were asymptomatic, 29.4% reported mild symptoms, 43.9% moderate symptoms and 3.7% severe symptoms. Three patients were hospitalized and discharged after recovery. Our results indicate that the use of telemedicine with diagnosis and drug treatment is a safe and effective strategy to reduce overload of health services and the exposure of healthcare providers and the population. The patients that initiated the treatment in the early stages of the disease presented satisfactory clinical response, reducing the need of face-to-face consultations and hospitalizations. The patients who followed the protocol treatment for COVID-19 with hydroxychloroquine and azithromycin for five days presented statistically significant improvement of clinical symptoms when compared to moderate patients who opted for not following the protocol (p < 0.05) and to all no treatment patients (p < 0.001).

Identification of B-Cell Linear Epitopes in the Nucleocapsid (N) Protein B-Cell Linear Epitopes Conserved among the Main SARS-CoV-2 Variants.

The Nucleocapsid (N) protein is highlighted as the main target for COVID-19 diagnosis by antigen detection due to its abundance in circulation early during infection. However, the effects of the described mutations in the N protein epitopes and the efficacy of antigen testing across SARS-CoV-2 variants remain controversial and poorly understood. Here, we used immunoinformatics to identify five epitopes in the SARS-CoV-2 N protein (N(34-48), N(89-104), N(185-197), N(277-287), and N(378-390)) and validate their reactivity against samples from COVID-19 convalescent patients. All identified epitopes are fully conserved in the main SARS-CoV-2 variants and highly conserved with SARS-CoV. Moreover, the epitopes N(185-197) and N(277-287) are highly conserved with MERS-CoV, while the epitopes N(34-48), N(89-104), N(277-287), and N(378-390) are lowly conserved with common cold coronaviruses (229E, NL63, OC43, HKU1). These data are in accordance with the observed conservation of amino acids recognized by the antibodies 7R98, 7N0R, and 7CR5, which are conserved in the SARS-CoV-2 variants, SARS-CoV and MERS-CoV but lowly conserved in common cold coronaviruses. Therefore, we support the antigen tests as a scalable solution for the population-level diagnosis of SARS-CoV-2, but we highlight the need to verify the cross-reactivity of these tests against the common cold coronaviruses.