Gamma-delta T cells suppress microbial metabolites that activate striatal neurons and induce repetitive/compulsive behavior in mice.

Intestinal γδ T cells play an important role in shaping the gut microbiota, which is critical not only for maintaining intestinal homeostasis but also for controlling brain function and behavior. Here, we found that mice deficient for γδ T cells (γδ-/-) developed an abnormal pattern of repetitive/compulsive (R/C) behavior, which was dependent on the gut microbiota. Colonization of WT mice with γδ-/- microbiota induced R/C behavior whereas colonization of γδ-/- mice with WT microbiota abolished the R/C behavior. Moreover, γδ-/- mice had elevated levels of the microbial metabolite 3-phenylpropanoic acid in their cecum, which is a precursor to hippurate (HIP), a metabolite we found to be elevated in the CSF. HIP reaches the striatum and activates dopamine type 1 (D1R)-expressing neurons, leading to R/C behavior. Altogether, these data suggest that intestinal γδ T cells shape the gut microbiota and their metabolites and prevent dysfunctions of the striatum associated with behavior modulation.

The microbiome-gut-brain axis in nutritional neuroscience.

Emerging evidence is highlighting the microbiome as a key regulator of the effect of nutrition on gut-brain axis signaling. Nevertheless, it is not yet clear whether the impact of nutrition is moderating the microbiota-gut-brain interaction or if diet has a mediating role on microbiota composition and function to influence central nervous system function, brain phenotypes and behavior. Mechanistic evidence from cell-based in vitro studies, animal models and preclinical intervention studies are linking the gut microbiota to the effects of diet on brain function, but they have had limited translation to human intervention studies. While increasing evidence demonstrates the triangulating relationship between diet, microbiota, and brain function across the lifespan, future mechanistic and translational studies in the field of microbiota and nutritional neuroscience are warranted to inform potential strategies for prevention and management of several neurological, neurodevelopmental, neurodegenerative, and psychiatric disorders. This brief primer provides an overview of the most recent advances in the nutritional neuroscience - microbiome field, highlighting significant opportunities for future research.

A traveling SARS-CoV-2 laboratory as part of a pandemic response among vulnerable Brazilian populations.

BACKGROUND: Brazil has been dramatically hit by the SARS-CoV-2 pandemic and is a world leader in COVID-19 morbidity and mortality. Additionally, the largest country of Latin America has been a continuous source of SARS-CoV-2 variants and shows extraordinary variability of the pandemic strains probably related to the country´s outstanding position as a Latin American economical and transportation hub. Not all regions of the country show sufficient infrastructure for SARS-CoV-2 diagnosis and genotyping which can negatively impact the pandemic response. METHODS: Due to this reason and to disburden the diagnostic system of the inner São Paulo State, the Butantan Institute established the Mobile Laboratory (in Portuguese: LabMovel) for SARS-CoV-2 testing which started a trip of the most important "hotspots" of the most populous Brazilian region. The LabMovel initiated in two important cities of the State: Aparecida do Norte (an important religious center) and the Baixada Santista region which incorporates the port of Santos, the busiest in Latin America. The LabMovel was fully equipped with an automatized system for SARS-CoV-2 diagnosis and sequencing/genotyping. It also integrated the laboratory systems for patient records and results divulgation including in the Federal Brazilian Healthcare System. RESULTS: Currently,16,678 samples were tested, among them 1,217 from Aparecida and 4,564 from Baixada Santista. We tracked the delta introductio in the tested regions with its high diversification. The established mobile SARS-CoV-2 laboratory had a major impact on the Public Health System of the included cities including timely delivery of the results to the healthcare agents and the Federal Healthcare system, evaluation of the vaccination status of the positive individuals in the background of exponential vaccination process in Brazil and scientific and technological divulgation of the fieldwork to the most vulnerable populations. CONCLUSIONS: The SARS-CoV-2 pandemic has demonstrated worldwide the importance of science to fight against this viral agent and the LabMovel shows that it is possible to integrate researchers, clinicians, healthcare workers and patients to take rapid actions that can in fact mitigate this and other epidemiological situations.

Self-Renewal and Pluripotency in Osteosarcoma Stem Cells' Chemoresistance: Notch, Hedgehog, and Wnt/ß-Catenin Interplay with Embryonic Markers.

Osteosarcoma is a highly malignant bone tumor derived from mesenchymal cells that contains self-renewing cancer stem cells (CSCs), which are responsible for tumor progression and chemotherapy resistance. Understanding the signaling pathways that regulate CSC self-renewal and survival is crucial for developing effective therapies. The Notch, Hedgehog, and Wnt/ß-Catenin developmental pathways, which are essential for self-renewal and differentiation of normal stem cells, have been identified as important regulators of osteosarcoma CSCs and also in the resistance to anticancer therapies. Targeting these pathways and their interactions with embryonic markers and the tumor microenvironment may be a promising therapeutic strategy to overcome chemoresistance and improve the prognosis for osteosarcoma patients. This review focuses on the role of Notch, Hedgehog, and Wnt/ß-Catenin signaling in regulating CSC self-renewal, pluripotency, and chemoresistance, and their potential as targets for anti-cancer therapies. We also discuss the relevance of embryonic markers, including SOX-2, Oct-4, NANOG, and KLF4, in osteosarcoma CSCs and their association with the aforementioned signaling pathways in overcoming drug resistance.

Detection of potential functional variants based on systems-biology: the case of feed efficiency in beef cattle.

BACKGROUND: Potential functional variants (PFVs) can be defined as genetic variants responsible for a given phenotype. Ultimately, these are the best DNA markers for animal breeding and selection, especially for polygenic and complex phenotypes. Herein, we described the identification of PFVs for complex phenotypes (in this case, Feed Efficiency in beef cattle) using a systems-biology driven approach based on RNA-seq data from physiologically relevant organs. RESULTS: The systems-biology coupled with deep molecular phenotyping by RNA-seq of liver, muscle, hypothalamus, pituitary, and adrenal glands of animals with high and low feed efficiency (FE) measured by residual feed intake (RFI) identified 2,000,936 uniquely variants. Among them, 9986 variants were significantly associated with FE and only 78 had a high impact on protein expression and were considered as PFVs. A set of 169 significant uniquely variants were expressed in all five organs, however, only 27 variants had a moderate impact and none of them a had high impact on protein expression. These results provide evidence of tissue-specific effects of high-impact PFVs. The PFVs were enriched (FDR < 0.05) for processing and presentation of MHC Class I and II mediated antigens, which are an important part of the adaptive immune response. The experimental validation of these PFVs was demonstrated by the increased prediction accuracy for RFI using the weighted G matrix (ssGBLUP+wG; Acc = 0.10 and b = 0.48) obtained in the ssGWAS in comparison to the unweighted G matrix (ssGBLUP; Acc = 0.29 and b = 1.10). CONCLUSION: Here we identified PFVs for FE in beef cattle using a strategy based on systems-biology and deep molecular phenotyping. This approach has great potential to be used in genetic prediction programs, especially for polygenic phenotypes.

Hexokinase 2: The preferential target of trehalose-6-phosphate over hexokinase 1.

Cancer-related metabolic features are in part maintained by hexokinase 2 upregulation, which leads to high levels of glucose-6-phosphate (G6P) and is needed to provide energy and biomass to support rapid proliferation. Using a humanized model of the yeast Saccharomyces cerevisiae, we explored how human hexokinase 2 (HK2) behaves under different nutritional conditions. At high glucose levels, yeast presents aerobic glycolysis through a regulatory mechanism known as catabolic repression, which exerts a metabolic adaptation like the Warburg effect. At high glucose concentrations, HK2 did not translocate into the nucleus and was not able to shift the metabolism toward a highly glycolytic state, in contrast to the effect of yeast hexokinase 2 (Hxk2), which is a crucial protein for the control of aerobic glycolysis in S. cerevisiae. During the stationary phase, when glucose is exhausted, Hxk2 is shuttled out of the nucleus, ceasing catabolic repression. Cells harvested at this condition display low glucose consumption rates. However, glucose-starved cells expressing HK2 had an increased capacity to consume glucose. In those cells, HK2 localized to mitochondria, becoming insensitive to G6P inhibition. We also found that the sugar trehalose-6-phosphate (T6P) is a human HK2 inhibitor, like yeast Hxk2, but was not able to inhibit human HK1, the isoform that is ubiquitously expressed in almost all mammalian tissues. In contrast to G6P, T6P inhibited HK2 even when HK2 was associated with mitochondria. The binding of HK2 to mitochondria is crucial for cancer survival and proliferation. T6P was able to reduce the cell viability of tumor cells, although its toxicity was not impressive. This was expected as cell absorption of phosphorylated sugars is low, which might be counteracted using nanotechnology. Altogether, these data suggest that T6P may offer a new paradigm for cancer treatment based on specific inhibition of HK2.

SARS-COV-2 genomic monitoring in the state of São Paulo unveils two emerging AY.43 sublineages.

Delta VOC is highly diverse with more than 120 sublineages already described as of November 30, 2021. In this study, through active monitoring of circulating severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) variants in the state of São Paulo, southeast Brazil, we identified two emerging sublineages from the ancestral AY.43 strain which were classified as AY.43.1 and AY.43.2. These sublineages were defined by the following characteristic nonsynonymous mutations ORF1ab:A4133V and ORF3a:T14I for the AY.43.1 and ORF1ab:G1155C for the AY.43.2 and our analysis reveals that they might have a likely-Brazilian origin. Much is still unknown regarding their dissemination in the state of São Paulo and Brazil as well as their potential impact on the ongoing vaccination process. However, the results obtained in this study reinforce the importance of genomic surveillance activity for timely identification of emerging SARS-CoV-2 variants which can impact the ongoing SARS-CoV-2 vaccination and public health policies.

Diet and the microbiota - gut - brain-axis: a primer for clinical nutrition.

PURPOSE OF REVIEW: Diet is an essential modulator of the microbiota - gut - brain communication in health and disease. Consequently, diet-induced microbiome states can impact brain health and behaviour. The integration of microbiome into clinical nutrition perspectives of brain health is sparse. This review will thus focus on emerging evidence of microbiome-targeted dietary approaches with the potential to improve brain disorders. RECENT FINDINGS: Research in this field is evolving toward randomized controlled trials using dietary interventions with the potential to modulate pathways of the microbiota - gut - brain-axis. Although most studies included small cohorts, the beneficial effects of Mediterranean-like diets on symptoms of depression or fermented foods on the immune function of healthy individuals shed light on how this research line can grow. With a clinical nutrition lens, we highlight several methodological limitations and knowledge gaps, including the quality of dietary intake information, the design of dietary interventions, and missing behavioural outcomes. SUMMARY: Findings in diet - microbiome - brain studies can have groundbreaking implications in clinical nutrition practice and research. Modulating brain processes through diet via the gut microbiota raises numerous possibilities. Novel dietary interventions targeting the microbiota - gut - brain-axis can offer various options to prevent and treat health problems such as mental disorders. Furthermore, knowledge in this field will improve current nutritional guidelines for disease prevention.

What 'Omics can tell us about antifungal adaptation.

Invasive candidiasis, the most frequent healthcare-associated invasive fungal infection, is commonly caused by Candida albicans. However, in recent years other antifungal-resistant Candida species-namely Candida glabrata and Candidaauris-have emerged as a serious matter of concern. Much of our understanding of the mechanisms regulating antifungal resistance and tolerance relies on studies utilizing C. albicans, C. glabrataand the model yeast Saccharomyces cerevisiae. 'Omics studies have been used to describe alterations in metabolic, genomic and transcriptomic expression profiles upon antifungal treatment of fungal cells. The physiological changes identified by these approaches could significantly affect fungal fitness in the host and survival during antifungal challenge, as well as provide further understanding of clinical resistance. Thus, this review aims to comparatively address 'omics data for C. albicans, C. glabrata andS. cerevisiae published from 2000 to 2021 to identify what these technologies can tell us regarding cellular responses to antifungal therapy. We will also highlight possible effects on pathogen survival and identify future avenues for antifungal research.

A prospective observational study for a Federated Artificial Intelligence solution for moniToring mental Health status after cancer treatment (FAITH): study protocol.

BACKGROUND: Depression is a common condition among cancer patients, across several points in the disease trajectory. Although presenting higher prevalence rates than the general population, it is often not reported or remains unnoticed. Moreover, somatic symptoms of depression are common in the oncological context and should not be dismissed as a general symptom of cancer. It becomes even more challenging to track psychological distress in the period after the treatment, where connection with the healthcare system typically becomes sporadic. The main goal of the FAITH project is to remotely identify and predict depressive symptoms in cancer survivors, based on a federated machine learning (ML) approach, towards optimization of privacy. METHODS: FAITH will remotely analyse depression markers, predicting their negative trends. These markers will be treated in distinct categories, namely nutrition, sleep, activity and voice, assessed in part through wearable technologies. The study will include 300 patients who have had a previous diagnosis of breast or lung cancer and will be recruited 1 to 5 years after the end of primary cancer. The study will be organized as a 12-month longitudinal prospective observational cohort study, with monthly assessments to evaluate depression symptoms and quality of life among cancer survivors. The primary endpoint is the severity of depressive symptoms as measured by the Hamilton Depression Rating Scale (Ham-D) at months 3, 6, 9 and 12. Secondary outcomes include self-reported anxiety and depression symptoms (HADS scale), and perceived quality of life (EORTC questionnaires), at baseline and monthly. Based on the predictive models gathered during the study, FAITH will also aim at further developing a conceptual federated learning framework, enabling to build machine learning models for the prediction and monitoring of depression without direct access to user's personal data. DISCUSSION: Improvements in the objectivity of psychiatric assessment are necessary. Wearable technologies can provide potential indicators of depression and anxiety and be used for biofeedback. If the FAITH application is effective, it will provide healthcare systems with a novel and innovative method to screen depressive symptoms in oncological settings. TRIAL REGISTRATION: Trial ID: ISRCTN10423782 . Date registered: 21/03/2022.

Scale of pregnant women's assessment of knowledge, attitude and practice related to urinary incontinence.

INTRODUCTION AND HYPOTHESIS: Urinary incontinence (UI) is prevalent during pregnancy and negatively affects quality of life. Despite this, few women seek professional assistance during prenatal care. Assessing the knowledge, attitude and practice (KAP) of pregnant women related to UI can contribute to the development and improvement of the quality of interventions performed during this period. For this, it is essential to apply targeted and valid instruments for this population. Thus, the present study aimed to construct and validate the content of a scale to evaluate the KAP of pregnant women related to UI. METHODS: Extensive literature review guided the operationalization of the instrument's initial items. Ten experts were selected for the theoretical analysis of the items, which was carried out using the Delphi technique, and the analysis of semantic adequacy proceeded from the application of the scale to 30 pregnant women. The data were analyzed using the content validity coefficient and kappa coefficient. RESULTS: The proposed Scale of Pregnant Women's Assessment of Knowledge, Attitude and Practice related to UI was approved by consensus by the experts, with a mean Cohen's kappa of 0.84 (p < 0.01), comprising 23 items. CONCLUSIONS: The results of the study confirm that the presented scale can be used as a valid tool to assess the KAP of pregnant women related to UI.

Evaluating aquatic macrophytes for removing erythromycin from contaminated water: floating or submerged?

Water contamination by antibiotics is an emerging global problem, with impacts on both public health and the environment. Erythromycin has been encountered in bodies of water throughout the world, which demands the development of efficient remediation technologies. We investigated the physiological responses and phytoremediation capacity of four species of aquatic macrophytes, two floating (Salvinia molesta and Lemna minor) and two submerged (Myriophyllum aquaticum and Rotala rotundifolia). The plants were exposed to relevant environmental concentrations of erythromycin (0 and 1.7 µg l-1) in artificially contaminated water for seven days. Physiological evaluations evidenced the ability of that antibiotic to promote oxidative events in those plants, such as the activation of antioxidant enzymes (ascorbate peroxidase and/or catalase). S. molesta exposed to erythromycin demonstrated accumulations of hydrogen peroxide and oxidative damage (lipid peroxidation) that was reflected in growth reductions. The erythromycin removal efficiency of floating plants varied from 9 to 12%, while submerged species varied from 31 to 44%. As such, submerged macrophyte species demonstrated the most efficient removal of erythromycin from contaminated waters, and are therefore more indicated for antibiotic phytoremediation projects.

For the very first time, the capacities of floating and submerged plant species used for removing erythromycin from contaminated water were compared. Moreover, plant physiological responses were related to their phytoremediation capacity. Our results promise to have direct impacts on plant and environmental science as well as in toxicology since they will contribute to a better understanding of the effects of antibiotics in plants and indicate species for better performance of phytoremediation programs aiming to reclaim the antibiotic erythromycin.

Chemoresistance-Related Stem Cell Signaling in Osteosarcoma and Its Plausible Contribution to Poor Therapeutic Response: A Discussion That Still Matters.

Osteosarcoma is amongst the most prevalent bone sarcomas and majorly afflicts children and adolescents. Therapeutic regimens based on the triad of doxorubicin, cisplatin and methotrexate have been used as the state-of-the-art approach to clinical treatment and management, with no significant improvements in the general outcomes since their inception in the early 1970s. This fact raises the following problematic questions: Why do some patients still relapse despite an initial good response to therapy? Why do nearly 30% of patients not respond to neoadjuvant therapies? Does residual persistent disease contribute to relapses and possible metastatic dissemination? Accumulating evidence suggests that chemoresistant cancer stem cells may be the major culprits contributing to those challenging clinical outcomes. Herein, we revisit the maneuvers that cancer stem cells devise for eluding cell killing by the classic cytotoxic therapies used in osteosarcoma, highlighting studies that demonstrate the complex crosstalk of signaling pathways that cancer stem cells can recruit to become chemoresistant.

[Combined use of serologic markers and spatial analysis for epidemiological surveillance of leprosyUso conjunto de los marcadores serológicos y del análisis espacial en la vigilancia epidemiológica de la lepra]. / Uso combinado de marcadores sorológicos e análise espacial na vigilância epidemiológica da hanseníase.

OBJECTIVE: To evaluate the combined use of serologic markers and spatial analysis to increase the sensitivity of leprosy epidemiological surveillance. METHOD: This cross-sectional study was performed with neighbors of leprosy cases and neighbors and family members of schoolchildren with a positive anti-phenolic glycolipid I (PGL-I) test in Diamantina, state of Minas Gerais, Brazil. Neighbors were those living within a 100-m radius of schoolchildren or leprosy cases. Data collection included a semi-structured interview, dermatologic examination, and rapid ML Flow test. All residential addresses were georeferenced. Multivariate regression and spatial analysis were performed with anti-PGL-I seropositivity as the dependent variable. RESULTS: The study included 1 491 individuals: 1 009 (67.7%) family members and neighbors of schoolchildren with positive serology test and 482 (32.3%) neighbors of leprosy cases. Of the total, 421 (28.2%) were positive for anti-PGL-I. Seropositivity was higher among family members and neighbors of seropositive schoolers (P < 0.001), among people with family income of 1 minimum wage (P < 0.001), among the youngest participants (P < 0.001), and among those living in homes with one to five rooms (P = 0.007). The seropositivity rate was higher in the geographic area corresponding to seropositive schoolchildren (P < 0.001), that is, the spot with the highest number of cases was different from the spot with the highest rate of seropositivity. CONCLUSIONS: The combined use of serologic markers and spatial analysis allowed us to easily identify operational weaknesses of services and a possible occult leprosy endemism in the municipality's urban census tracts. Tracing of social contact and neighbors, active search, educational campaigns, school surveys, and territorial analyses facilitate the early diagnosis of leprosy.

OBJETIVO: Evaluar el uso conjunto de los marcadores serológicos y del análisis espacial para ampliar la sensibilidad de la vigilancia epidemiológica de la lepra. MÉTODO: Este estudio transversal se realizó con vecinos de personas con casos de lepra y con familiares y vecinos de escolares con resultados positivos en las pruebas serológicas del antígeno glicolípido fenólico I (PGL-I) en Diamantina (Minas Gerais, Brasil). Se definieron como vecinos las personas residentes en un radio de hasta 100 metros de distancia de los escolares y de las personas con casos de lepra. Para la recopilación de datos se realizaron entrevistas semiestructuradas, exámenes dermatoneurológicos y pruebas serológicas rápidas ML Flow. Todas las direcciones se ubicaron por georreferencia. Se realizó una regresión multivariante y un análisis espacial, y se empleó la reactividad serológica anti-PGL-I como variable dependiente. RESULTADOS: Se estudiaron 1 491 personas: 1 009 (67,7%) familiares y vecinos de escolares con resultados positivos en las pruebas serológicas y 482 (32,3%) vecinos de las personas con casos de lepra. Del total, 421 (28,2%) presentaron seropositividad anti-PGL-I. La posibilidad de presentar seropositividad fue mayor en los familiares y vecinos de los escolares seropositivos (P < 0,001), en las personas con ingresos familiares de un salario mínimo (P < 0,001), en las personas más jóvenes (P < 0,001) y en los residentes en domicilios con una a cinco habitaciones (P = 0,007). La tasa de seropositividad fue mayor en la zona geográfica correspondiente a los escolares seropositivos (P < 0,001), por lo que hubo divergencia entre el foco de mayor concentración de casos y el de mayor seropositividad. CONCLUSIONES: El uso conjunto de los marcadores serológicos y del análisis espacial permitió determinar las fragilidades operativas de los servicios y una posible endemia oculta de lepra en los sectores urbanos del municipio incluidos en el censo correspondiente. Las actividades de rastreo de contactos sociales y cercanos, la búsqueda activa de casos, las campañas educativas, las encuestas escolares y el análisis del territorio facilitan el diagnóstico temprano de la lepra.

Knowledge, attitude, and practice instruments for urinary incontinence: A psychometric review.

AIMS: To analyze the psychometric properties of instruments measure knowledge and/or attitude and/or practice related to urinary incontinence (UI). METHODS: This review was conducted in September 2018 without restrictions regarding language and year of publication. DATA SOURCES: MEDLINE/PubMed, CINAHL, SCOPUS, PsycINFO, Web of Science, Cochrane, LILACS, and Google. The psychometric properties were evaluated using the COnsensus-based Standards for the selection of health status Measurement INstruments and the Quality Criteria for Measurement Properties. RESULTS: A total of seven articles published between 1999 and 2018 were included. The evaluation of the psychometric properties showed that the majority of the instruments presented "excellent" or "good" methodological quality; however, the description of aspects related to the validation stages was inadequate in some studies, causing the quality of the results to be evaluated as "indeterminate" in some properties. This review cannot recommend any questionnaire without reservations; however, the Urinary Incontinence Scales and the Barriers Incontinence Care Seeking Questionnaire presented better evaluations, regarding both methodological quality and the quality of the results. CONCLUSION: This review provides a strategy to select the most suitable instrument to assess knowledge, attitude, and practice regarding UI, according to the psychometric properties, guiding their use by researchers in clinical practice.

Wastewater Chlorination for Reuse, an Alternative for Small Communities.

Reuse of wastewater is a practice that has been employed all over the world, mainly in agriculture, where the main aim is to reduce the demand for water and provide nutrients. However, these waters for reuse often have excessive amounts of pathogenic microorganisms, requiring a specific disinfection step even after being subjected to a purification process. Thus, this research aimed to evaluate the potential use of calcium hypochlorite and sodium hypochlorite as disinfectant agents for the sanitary effluent of the treatment system based on constructed wetlands for later reuse. Disinfection tests were carried out in batch, using three dosages of hypochlorite (5, 10 and 15 mg.L-1) and different contact. In all disinfection tests, inactivation of indicator microorganisms (total coliforms and E. coli) was considered effective for the two disinfectant agents adopted, satisfying the criteria for reuse according to the World Health Organization (WHO). There was no formation of trihalomethanes after disinfection tests.

Psychometric properties of the Portuguese version of the Yale Food Addiction Scale.

PURPOSE: Rising rates of obesity have been recently associated to the novel concept of food addiction (FA). The Yale Food Addiction Scale (YFAS) is the most widely used measure for examining FA (1) and analysis of its reliability and validity is expected to facilitate empirical research on the construct. Here, we tested the psychometric properties of a Portuguese version of the YFAS (P-YFAS), establishing its factor structure, reliability and construct validity. METHODS: Data were obtained from 468 Portuguese individuals, 278 sampled from non-clinical populations, and 190 among obese candidates for weight-loss surgery. A battery of self-report measures of eating behavior was applied. RESULTS: Confirmatory factor analysis verified a one-factor structure with acceptable fit, with item analysis suggesting the need to eliminate item 24 from the P-YFAS. Internal consistency (KR-20 = .82) and test-retest stability were adequate. Correlation analyses supported convergent and divergent validity of the P-YFAS, particularly in the clinical sample. Both FA symptom count and diagnosis, according to the P-YFAS, adequately discriminated between samples, with classification of FA met by 2.5 and 25.8% of the participants in the non-clinical and clinical samples, respectively. CONCLUSIONS: These findings reinforce the use of P-YFAS in non-clinical and clinical populations. Future directions for extending YFAS validation are discussed.

Pydna: a simulation and documentation tool for DNA assembly strategies using python.

BACKGROUND: Recent advances in synthetic biology have provided tools to efficiently construct complex DNA molecules which are an important part of many molecular biology and biotechnology projects. The planning of such constructs has traditionally been done manually using a DNA sequence editor which becomes error-prone as scale and complexity of the construction increase. A human-readable formal description of cloning and assembly strategies, which also allows for automatic computer simulation and verification, would therefore be a valuable tool. RESULTS: We have developed pydna, an extensible, free and open source Python library for simulating basic molecular biology DNA unit operations such as restriction digestion, ligation, PCR, primer design, Gibson assembly and homologous recombination. A cloning strategy expressed as a pydna script provides a description that is complete, unambiguous and stable. Execution of the script automatically yields the sequence of the final molecule(s) and that of any intermediate constructs. Pydna has been designed to be understandable for biologists with limited programming skills by providing interfaces that are semantically similar to the description of molecular biology unit operations found in literature. CONCLUSIONS: Pydna simplifies both the planning and sharing of cloning strategies and is especially useful for complex or combinatorial DNA molecule construction. An important difference compared to existing tools with similar goals is the use of Python instead of a specifically constructed language, providing a simulation environment that is more flexible and extensible by the user.

Optimizing Screw Speed and Barrel Temperature for Textural and Nutritional Improvement of Soy-Based High-Moisture Extrudates.

Soy remains the legume protein of excellence for plant-based meat alternatives due to its fiber-forming potential. In this study, protein-rich powders from soy protein isolate (SPI), concentrate (SPC), and their mixture (SPM) were thoroughly characterized for their proximate composition, nutritional quality, and physicochemical properties to understand their structuring behavior during high-moisture extrusion. SPI presented higher degrees of protein denaturation and aggregation, least gelation concentration and lower essential amino acid contents. Thus, an SPI:SPC combination (1:9 ratio, 70% protein) was extruded at three different screw speeds (300, 350, and 400 rpm) and two temperature profiles (120 and 140 °C maximum temperature). The effects of the processing parameters on the extrudates were evaluated for their appearance (fibrousness), texture (TPA, cutting force, and anisotropy), color, protein structure (FTIR), and trypsin inhibitors. Higher temperatures resulted in softer and darker extrudates, with increased visual and instrumental anisotropy. Increasing screw speeds led to softer and lighter extrudates, without a clear fibrousness effect. ß-sheet structures decreased and intermolecular aggregates (A1) increased after extrusion, especially at 140 °C, together with the formation of intramolecular aggregates (A2). Extrusion also significantly decreased the amount of trypsin inhibitors (>90%). This study demonstrates that extrusion parameters need to be carefully selected to achieve meat analogs with optimal textural and nutritional characteristics.

Cloud point method applied to the extraction and preconcentration of thiabendazole pesticide from whole grape juice samples and amperometric detection by HPLC.

A cloud point method was developed and applied for the first time to extract and preconcentrate thiabendazole (TBZ) from commercial whole grape juice samples, with determination by high performance liquid chromatography coupled to electrochemical detection (HPLC/EC), using a cathodically pretreated boron-doped diamond electrode (BDD). The best conditions for extraction and preconcentration of TBZ by cloud point extraction (CPE) were performed at pH 6.0, by adding 1 mL of the surfactant Tergitol TMN-6 at 10% (mass-to-mass ratio), without heating (at 27 °C) and ultrasonic stirring time of (20 kHz) for 60 min. The HPLC/EC determination was duly validated in a C8 column, in mobile phase with a 69 : 31 ratio (V/V) of phosphate buffer (pH 7.0):ACN, at a flow rate of 1.2 mL min-1 and electrochemical detection with BDD electrode by applying 1.40 V × Ag/AgCl (3.0 mol L-1). Under these conditions, the procedure showed a preconcentration factor (FC) of 21.7, and limits of detection (LOD) and quantification (LOQ) of 6.64 × 10-9 mol L-1 (or 1.33 µg L-1) and 1.66 × 10-8 mol L-1 (or 3.34 µg L-1), respectively. The method provided a percent recovery of 81% to 98%, with a coefficient of variation between 3% and 15%.