How different attributes are weighted in professionals' decision-making in Pediatric Dentistry-a protocol for guiding discrete choice experiment focused on shortening the evidence-based practice implementation for dental care.

BACKGROUND: Important evidence has been constantly produced and needs to be converted into practice. Professional consumption of such evidence may be a barrier to its implementation. Then, effective implementation of evidence-based interventions in clinical practice leans on the understanding of how professionals value attributes when choosing between options for dental care, permitting to guide this implementation process by maximizing strengthens and minimizing barriers related to that. METHODS: This is part of a broader project investigating the potential of incorporating scientific evidence into clinical practice and public policy recommendations and guidelines, identifying strengths and barriers in such an implementation process. The present research protocol comprises a Discrete Choice Experiment (DCE) from the Brazilian oral health professionals' perspective, aiming to assess how different factors are associated with professional decision-making in dental care, including the role of scientific evidence. Different choice sets will be developed, either focusing on understanding the role of scientific evidence in the professional decision-making process or on understanding specific attributes associated with different interventions recently tested in randomized clinical trials and available as newly produced scientific evidence to be used in clinical practice. DISCUSSION: Translating research into practice usually requires time and effort. Shortening this process may be useful for faster incorporation into clinical practice and beneficial to the population. Understanding the context and professionals' decision-making preferences is crucial to designing more effective implementation and/or educational initiatives. Ultimately, we expect to design an efficient implementation strategy that overcomes threats and potential opportunities identified during the DCEs, creating a customized structure for dental professionals. TRIAL REGISTRATION: https://osf.io/bhncv .

Novel SK channel positive modulators prevent ferroptosis and excitotoxicity in neuronal cells.

Small conductance calcium-activated potassium (SK) channel activity has been proposed to play a role in the pathology of several neurological diseases. Besides regulating plasma membrane excitability, SK channel activation provides neuroprotection against ferroptotic cell death by reducing mitochondrial Ca2+ uptake and reactive oxygen species (ROS). In this study, we employed a multifaceted approach, integrating structure-based and computational techniques, to strategically design and synthesize an innovative class of potent small-molecule SK2 channel modifiers through highly efficient multicomponent reactions (MCRs). The compounds' neuroprotective activity was compared with the well-studied SK positive modulator, CyPPA. Pharmacological SK channel activation by selected compounds confers neuroprotection against ferroptosis at low nanomolar ranges compared to CyPPA, that mediates protection at micromolar concentrations, as shown by an MTT assay, real-time cell impedance measurements and propidium iodide staining (PI). These novel compounds suppress increased mitochondrial ROS and Ca2+ level induced by ferroptosis inducer RSL3. Moreover, axonal degeneration was rescued by these novel SK channel activators in primary mouse neurons and they attenuated glutamate-induced neuronal excitability, as shown via microelectrode array. Meanwhile, functional afterhyperpolarization of the novel SK2 channel modulators was validated by electrophysiological measurements showing more current change induced by the novel modulators than the reference compound, CyPPA. These data support the notion that SK2 channel activation can represent a therapeutic target for brain diseases in which ferroptosis and excitotoxicity contribute to the pathology.

Lipid profile in breast cancer: From signaling pathways to treatment strategies.

Breast cancer is the most prevalent cancer in women. Metabolic abnormalities, particularly increased lipid synthesis and uptake, impact the onset and progression of the disease. However, the influence of lipid metabolism in breast cancer varies according to the disease stage and patient's hormone status. In postmenopausal patients, obesity is associated with a higher risk and poor prognosis of luminal tumors, while in premenopausal individuals, it is correlated to BRCA mutated tumors. In fact, the tumor's lipid profile may be used to distinguish between HER2+, luminal and BRCA-mutated tumors. Moreover, drug resistance was associated with increased fatty acid synthesis and alterations in membrane composition, impacting its fluidity and spatial subdomains such as lipid rafts. Here, we discuss the subtype-specific lipid metabolism alterations found in breast cancer and the potentiality of its modulation in a clinical setting.

Individual satisfaction with HIV/AIDS care in Belo Horizonte, Brazil.

Individual satisfaction is associated with increased retention in HIV/AIDS care services and adherence to treatment. This study assessed factors associated with individual satisfaction at the initiation of antiretroviral therapy and compared the proportions of satisfaction at the initiation of antiretroviral therapy and after three months of follow-up. Face-to-face interviews were carried out among 398 individuals of three HIV/ AIDS healthcare services in Belo Horizonte, Brazil. Variables included sociodemographic and clinical characteristics, perception about healthcare services, and domains of quality of life. Individuals that rated the quality of healthcare services as good or very good were classified as satisfied. A logistic regression analysis of the association between independent variables and individual satisfaction was performed. The proportion of individual satisfaction with healthcare services was 95.5% at the initiation of antiretroviral therapy and 96.7% after three months and these changes were not statistically significant (p = 0.472). The physical domain of quality of life was found to be associated with satisfaction at the initiation of antiretroviral therapy (OR = 1.38; CI = 1.11-1.71; p = 0.003). Training and supervision of health professionals for addressing the needs of individuals with lower levels of physical domain of quality of life may improve satisfaction with HIV/ AIDS care.

Negative modulation of mitochondrial calcium uniporter complex protects neurons against ferroptosis.

Ferroptosis is an iron- and reactive oxygen species (ROS)-dependent form of regulated cell death, that has been implicated in Alzheimer's disease and Parkinson's disease. Inhibition of cystine/glutamate antiporter could lead to mitochondrial fragmentation, mitochondrial calcium ([Ca2+]m) overload, increased mitochondrial ROS production, disruption of the mitochondrial membrane potential (ΔΨm), and ferroptotic cell death. The observation that mitochondrial dysfunction is a characteristic of ferroptosis makes preservation of mitochondrial function a potential therapeutic option for diseases associated with ferroptotic cell death. Mitochondrial calcium levels are controlled via the mitochondrial calcium uniporter (MCU), the main entry point of Ca2+ into the mitochondrial matrix. Therefore, we have hypothesized that negative modulation of MCU complex may confer protection against ferroptosis. Here we evaluated whether the known negative modulators of MCU complex, ruthenium red (RR), its derivative Ru265, mitoxantrone (MX), and MCU-i4 can prevent mitochondrial dysfunction and ferroptotic cell death. These compounds mediated protection in HT22 cells, in human dopaminergic neurons and mouse primary cortical neurons against ferroptotic cell death. Depletion of MICU1, a [Ca2+]m gatekeeper, demonstrated that MICU is protective against ferroptosis. Taken together, our results reveal that negative modulation of MCU complex represents a therapeutic option to prevent degenerative conditions, in which ferroptosis is central to the progression of these pathologies.

The perspectives of pharmacy student preceptors on a service-learning program in primary health care: A qualitative study.

Background: There are a few studies about preceptorship in primary health care settings, but they didn't assess all possible contributions of this experience, such as preceptor behavioral changes and impact on health services. Objective: This study aimed to analyze the perspective of pharmacy student preceptors on a service-learning program carried out in a primary health care setting. Methods: This was a qualitative descriptive study, with semi-structured interviews and thematic analysis. The themes were defined deductively, according to levels of Kirkpatrick's theory about the impact of educational activities. Results: The preceptors showed positive and negative reactions to the structure of the service-learning program, developed their knowledge, changed behaviors such as collaborative interprofessional practice, and reported improvements in the health units. Conclusions: From the perspective of pharmacy student preceptor preceptors, the service-learning program had an impact on all levels expected for an educational activity according to Kirkpatrick's theory: reaction, learning, behavior, and results. These findings may inform the design of service-learning programs.

Species-specific metabolic reprogramming in human and mouse microglia during inflammatory pathway induction.

Metabolic reprogramming is a hallmark of the immune cells in response to inflammatory stimuli. This metabolic process involves a switch from oxidative phosphorylation (OXPHOS) to glycolysis or alterations in other metabolic pathways. However, most of the experimental findings have been acquired in murine immune cells, and little is known about the metabolic reprogramming of human microglia. In this study, we investigate the transcriptomic, proteomic, and metabolic profiles of mouse and iPSC-derived human microglia challenged with the TLR4 agonist LPS. We demonstrate that both species display a metabolic shift and an overall increased glycolytic gene signature in response to LPS treatment. The metabolic reprogramming is characterized by the upregulation of hexokinases in mouse microglia and phosphofructokinases in human microglia. This study provides a direct comparison of metabolism between mouse and human microglia, highlighting the species-specific pathways involved in immunometabolism and the importance of considering these differences in translational research.

Exogenous OCT4 and SOX2 Contribution to In Vitro Reprogramming in Cattle.

Mechanisms of cell reprogramming by pluripotency-related transcription factors or nuclear transfer seem to be mediated by similar pathways, and the study of the contribution of OCT4 and SOX2 in both processes may help elucidate the mechanisms responsible for pluripotency. Bovine fibroblasts expressing exogenous OCT4 or SOX2, or both, were analyzed regarding the expression of pluripotency factors and imprinted genes H19 and IGF2R, and used for in vitro reprogramming. The expression of the H19 gene was increased in the control sorted group, and putative iPSC-like cells were obtained when cells were not submitted to cell sorting. When sorted cells expressing OCT4, SOX2, or none (control) were used as donor cells for somatic cell nuclear transfer, fusion rates were 60.0% vs. 64.95% and 70.53% vs. 67.24% for SOX2 vs. control and OCT4 vs. control groups, respectively; cleavage rates were 66.66% vs. 81.68% and 86.47% vs. 85.18%, respectively; blastocyst rates were 33.05% vs. 44.15% and 52.06% vs. 44.78%, respectively. These results show that the production of embryos by NT resulted in similar rates of in vitro developmental competence compared to control cells regardless of different profiles of pluripotency-related gene expression presented by donor cells; however, induced reprogramming was compromised after cell sorting.

The Top 100 Most-Cited Articles on Permanent Teeth Avulsion: A Bibliometric Analysis.

INTRODUCTION: Knowledge of bibliometric indices on the avulsion of permanent teeth can help researchers to identify gaps in the literature. The aim of this study was to analyze the bibliometric characteristics of the 100 most-cited articles on the avulsion of permanent teeth. MATERIALS AND METHODS: Two researchers conducted an unfiltered search in the Core Collection section of the Web of Science (WoS-CC) database. The following data were extracted: position in the citation list, total number and density of citations, title, authors, institution, country, year of publication, journal, keywords, study design, tooth storage medium, and other topics of interest. Bibliometric networks were generated and analyzed using the VOSviewer software. Descriptive analysis and a Poisson regression analysis were performed (P < .05). RESULTS: The number of citations per article varied between 31 and 407. The average was 70.05, and 13 articles received more than 100 citations. J.O. Andreasen (n = 13) and M. Trope (n = 15) were the authors who contributed most often to publications. The United States was the country with the highest number of articles (n = 31). Authors associated with the University of Copenhagen, Denmark (n = 9) and University of North Carolina, USA (n = 10) showed higher productivity. In vitro/animal model studies were the most frequent study designs (n = 50), and the most-discussed topic of interest was prognosis (n = 47). There was strong clustering among 13 research groups. CONCLUSION: Observational studies, published by authors from Europe and about prognosis or treatment presented more citation ratio in WoS-CC. Clinical studies on permanent tooth avulsion are needed to increase the quality of scientific evidence and provide insights for health professionals and public policy makers.

Mitochondrial transplantation rescues neuronal cells from ferroptosis.

Ferroptosis is a type of oxidative cell death that can occur in neurodegenerative diseases and involves damage to mitochondria. Previous studies demonstrated that preventing mitochondrial dysfunction can rescue cells from ferroptotic cell death. However, the complexity of mitochondrial dysfunction and the timing of therapeutic interventions make it difficult to develop an effective treatment strategy against ferroptosis in neurodegeneration conditions. In this study, we explored the use of mitochondrial transplantation as a novel therapeutic approach for preventing ferroptotic neuronal cell death. Our data showed that isolated exogenous mitochondria were incorporated into both healthy and ferroptotic immortalized hippocampal HT-22 cells and primary cortical neurons (PCN). The mitochondrial incorporation was accompanied by increased metabolic activity and cell survival through attenuating lipid peroxidation and mitochondrial superoxide production. Further, the function of mitochondrial complexes I, III and V activities contributed to the neuroprotective activity of exogenous mitochondria. Similarly, we have also showed the internalization of exogenous mitochondria in mouse PCN; these internalized mitochondria were found to effectively preserve the neuronal networks when challenged with ferroptotic stimuli. The administration of exogenous mitochondria into the axonal compartment of a two-compartment microfluidic device induced mitochondrial transportation to the cell body, which prevented fragmentation of the neuronal network in ferroptotic PCN. These findings suggest that mitochondria transplantation may be a promising therapeutic approach for protecting neuronal cells from ferroptotic cell death.

Is there an association between dental caries and genetics? Systematic review and meta-analysis of studies with twins.

OBJECTIVE: This review aimed to assess the agreement of dental caries experience between monozygotic (MZ) and dizygotic (DZ) twins. DATA RESOURCES: This systematic review was performed by reviewers in the databases Embase, MEDLINE-PubMed, Scopus, Web of Science and manual searches and gray literature Google Scholar® and Opengray. Observational studies that evaluated dental caries in twins were included. The risk of bias was analyzed using the Joanna Briggs checklist. Meta-analyses were performed to assess the pooled Odds Ratio to estimate the agreement values of dental caries experience and DMF index between pairs of twins (p < 0.05). To assess the certainty of evidence, the GRADE scale was used. STUDY SELECTION: 2533 studies were identified, of which 19 were included in the qualitative analysis, six in the quantitative synthesis, with two meta-analyses being carried out. Association between genetics and the development of the disease was observed in most studies. In the risk of bias analysis, 47.4% had moderate risk. Higher agreement of dental caries experience was observed in MZ twins than DZ in both dentitions (OR: 5.94; 95% CI: 2.00-17.57). However, there was no difference between the MZ and DZ twin groups in the analysis comparing DMF index agreement (OR: 2.86; 95%CI: 0.25-32.79). The certainty of evidence was considered low and very low for all studies included in meta-analyses. CONCLUSION: With very low certainty of the evidence, the genetic factor seems to influence the agreement of the caries experience. CLINICAL RELEVANCE: Understanding the genetic influence on the disease has the potential to contribute to the development of studies that may use biotechnologies for prevention and treatment as well as guide future research involving gene therapies aiming to prevent the occurrence of dental caries.

Association between genetic factors and molar-incisor hypomineralisation or hypomineralised second primary molar: A systematic review.

OBJECTIVE: To determine the association between genetic factors and molar-incisor hypomineralisation (MIH) and/or hypomineralised second primary molars by means of a systematic review. DESIGN: A search was performed in Medline-PubMed, Scopus, Embase and Web of Science databases; manual search and search in gray literature were also performed. Selection of articles was performed independently by two researchers. A third examiner was involved in cases of disagreement. Data extraction was performed using an Excel® spreadsheet and independent analysis was performed for each outcome. RESULTS: Sixteen studies were included. There was an association between MIH and genetic variants related to amelogenesis, immune response, xenobiotic detoxification and other genes. Moreover, interactions between amelogenesis and immune response genes, and SNPs in the aquaporin gene and vitamin D receptors were associated with MIH. Greater agreement of MIH was found in pairs of monozygotic twins than dizygotic twins. The heritability of MIH was 20 %. Hypomineralised second primary molars was associated with SNPs in the hypoxia-related HIF-1 gene and methylation in genes related to amelogenesis. CONCLUSION: With very low or low certainty of evidence, an association was observed between MIH and SNPs in genes associated with amelogenesis, immune response, xenobiotic detox and ion transport. Interactions between genes related to amelogenesis and immune response as well as aquaporin genes were associated to MIH. With very low certainty of evidence, hypomineralised second primary molars was associated to a hypoxia-related gene and to methylation in genes related to amelogenesis. Moreover, higher agreement of MIH in pairs of monozygotic twins than dizygotic twins was observed.

Risk assessment validation in patients with pulmonary arterial hypertension: Data from a Southern Brazil registry (RESPHIRAR study).

Pulmonary arterial hypertension (PAH) is a severe and progressive disease characterized by increased pulmonary vascular resistance, ultimately leading to right heart failure and death. Registries are a valuable tool in the research of rare conditions such as PAH. Moreover, the risk assessment strategy has been validated in European and North American registries and has been reported to provide an accurate prediction of mortality and the clinical advantage of reaching low-risk status. However, there is no available data from Brazil. Thus, the aim of the present study was to describe the characteristics of a sample of PAH from Southern Brazil and to retrospectively validate the risk assessment at our population. The RESPHIRAR is a retrospective and multicentric registry on pulmonary hypertension. With a join collaboration from nine centers in Southern Brazil, demographics, clinical presentation, and hemodynamics data of PAH were collected between 2007 and 2017. Moreover, the REVEAL 2.0 and REVEAL 2.0 Lite risk assessments were validated in our population. Overall, 370 PAH patients were included in the present study. Patients were predominantly female (78.5%) and had a mean age of 41.8 ± 18.8 years. Most patients (33.4%) had idiopathic PAH, 30.2% had PAH associated with congenital heart disease, and 23.5% had PAH associated with connective tissue disease. The low-risk group showed significantly lower mortality than the intermediated- or high-risk group at diagnosis (p < 0.05). In conclusion, our data suggest that REVEAL 2.0 and REVEAL 2.0 Lite risk assessments can predict mortality risk in PAH patients in Southern Brazil.

Prepubertal to adulthood exposure to low doses of glyphosate-based herbicide increases the expression of the Havcr1 (Kim1) biomarker and causes mild kidney alterations.

Glyphosate is a nonselective and postemergent herbicide used to combat weeds in several crops, which raises concerns about risks to human health since residues are detected in urine, human milk, surface water and several types of food. Feces and urine are the major routes of elimination of glyphosate, making the kidney a sensitive target for the development of toxicity. In fact, farmers are at high risk of developing chronic kidney disease. In this sense, this study aims to investigate kidney function by measuring the serum levels of urea and creatinine, examining the histological morphology, and analyzing the mRNA expression of genes related to tubular transport of ions, urea and urates and the biomarker of kidney disease Kim1, and the levels of lead in the kidney in male Wistar rats orally exposed to low levels of glyphosate-based herbicide (GBH: 0, 0.5 or 5 mg/kg) from weaning to adult life by gavage. GBH0.5 showed reduced serum urea concentration, presence of tubulointerstitial swelling and mononuclear cell infiltration into the interstitium, increased gene expression of Kim1 and reduced gene expression of Slc14a1. GBH5 showed reduced serum urea and increased serum creatinine concentrations, tubulointerstitial swelling, interstitial fibrosis, and reduced expression of Trpm6 and Trpv5. Exposure to GBH did not affect the levels of Pb in the kidneys of animals. In conclusion, glyphosate at low doses may cause mild kidney damage. It is necessary to evaluate whether the long-term effects of this constant injury may contribute to the development of chronic kidney disease of uncertain etiology.

Mitochondrial dysfunction mediates neuronal cell response to DMMB photodynamic therapy.

Photodynamic therapy (PDT) is a process in which a photosensitizer (PS) is exposed to specific wavelengths and generates reactive oxygen species (ROS) which act within nanometers. The low invasive nature and directed cytotoxicity of this approach render it attractive to the treatment of different conditions, including the ones that affect the central nervous system (CNS). The effect of PDT on healthy neurons is one main concern over its use in the CNS, since neuronal-like cells were shown to be particularly sensitive to certain PSs. Among available PSs, 1,9-dimethyl-methylene blue (DMMB) stands out as being resistant to reduction to its inactive leuco form and by being able to produce high levels of singlet­oxygen. In this study, we aimed to investigate DMMB photodamage mechanisms in the hippocampal cell line HT22. Our results demonstrate that DMMB-PDT decrease in cell viability was linked with an increase in cell death and overall ROS production. Besides, it resulted in a significant increase in mitochondrial ROS production and decreased mitochondria membrane potential. Furthermore, DMMB-PDT significantly increased the presence of acidic autolysosomes, which was accompanied by an increase in ATG1 and ATG8 homologue GaBarap1 expression, and decreased DRAM1 expression. Taken together our results indicated that mitochondrial and autophagic dysfunction underlie DMMB-PDT cytotoxicity in neuronal cells.

Characterization of Lipopolysaccharide Effects on LRRK2 Signaling in RAW Macrophages.

Dysfunction of the immune system and mitochondrial metabolism has been associated with Parkinson's disease (PD) pathology. Mutations and increased kinase activity of leucine-rich repeat kinase 2 (LRRK2) are linked to both idiopathic and familial PD. However, the function of LRRK2 in the immune cells under inflammatory conditions is contradictory. Our results showed that lipopolysaccharide (LPS) stimulation increased the kinase activity of LRRK2 in parental RAW 264.7 (WT) cells. In addition to this, LRRK2 deletion in LRRK2 KO RAW 264.7 (KO) cells altered cell morphology following LPS stimulation compared to the WT cells, as shown by an increase in the cell impedance as observed by the xCELLigence measurements. LPS stimulation caused an increase in the cellular reactive oxygen species (ROS) levels in both WT and KO cells. However, WT cells displayed a higher ROS level compared to the KO cells. Moreover, LRRK2 deletion led to a reduction in interleukin-6 (IL-6) inflammatory cytokine and cyclooxygenase-2 (COX-2) expression and an increase in lactate production after LPS stimulation compared to the WT cells. These data illustrate that LRRK2 has an effect on inflammatory processes in RAW macrophages upon LPS stimulation.

Lesion sterilization and tissue repair with chloramphenicol, tetracyline, zinc oxide/eugenol paste versus conventional pulpectomy: A 36-month randomized controlled trial.

BACKGROUND: Endodontic treatment of primary molars represents one of the challenges in pediatric dentistry. There is a lack of consensus in the literature about the endodontic techniques and filling paste for primary teeth with pulp necrosis. AIM: To compare the effectiveness of the LSTR technique (lesion sterilization and tissue repair) with CTZ paste (chloramphenicol, tetracycline, zinc oxide and eugenol) and pulpectomy with ZOE paste (zinc oxide and eugenol) in the treatment of primary molars with pulp necrosis. DESIGN: Eighty-eight primary molars with pulp necrosis from 70 children between the ages of 3 and 8 years were included. The teeth were randomized to the LSTR with CTZ paste group or pulpectomy with ZOE paste group. Clinical and radiographic evaluations were performed at 18, 24, 30 and 36 months. RESULTS: At 36 months, clinical success was 86.4% in LSTR with CTZ paste and 90.9% in pulpectomy with ZOE paste (p = .45). Radiographic success was 43.2% in both groups (p = 1.00). The overall success was 40.9% in LSTR with CTZ paste and 43.2% in pulpectomy with ZOE paste (p = 1.00). CONCLUSION: After 36 months of evaluation, the effectiveness of the LSTR technique with CTZ paste and pulpectomy with ZOE paste was similar for the treatment of primary molars with pulp necrosis.

Chemical structure and biological activity of the (1 → 3)-linked ß-D-glucan isolated from marine diatom Conticribra weissflogii.

Several polysaccharides are considered to be "biological response modifiers" (BRM) - these refer to biomolecules that augment immune responses and can be derived from a variety of sources. Microalgae produce a diverse range of polysaccharides and could be an excellent source of BRM. Here, we describe the chemical structure and biological activity of water-soluble polysaccharide isolated from the marine diatom Conticribra weissflogii. Using chemical and NMR spectroscopic methods, the polysaccharide was identified as a (1 â†’ 3)-linked ß-D-glucan with a low proportion of C-6 substitution by single ß-glucose units. The biological activity of this low molecular weight ß-glucan (11.7 kDa) was investigated with respect to glioblastoma cell lines (U87 MG and U251) and macrophages (RAW 264.7). We observed that this ß-D-glucan did not exhibit cytotoxic activity against glioblastoma cells, but did enhance the phagocytic activity of macrophages, suggesting that it possesses immunomodulatory properties.

Asthma control in Brazil: a systematic review.

OBJECTIVES: To explore asthma control in patients undergoing pharmacotherapy on studies in the last 20 years in Brazil. Asthma is a chronic airway inflammation disease with a high prevalence worldwide. Even with a variety of drug treatment improvements, attaining asthma control is challenging, since it should have a personalized approach. In Brazil, studies on the prevalence of asthma control are scarce and usually from a small sample size. DATA SOURCES: A systematic review was performed to assess asthma control in Brazilian population. Terms related to "asthma", "asthma control" and "Brazil" were used in the search strategies in PubMed, BVSalud, Embase and Cochrane Library, including Brazilian Journal of Allergy and Immunology as data sources. A narrative synthesis was performed to report key outcome. STUDY SELECTIONS: In total, 23 studies were included. Most of them were conducted in the Southeastern and Northeast regions, in a short duration. RESULTS: Pediatric and non-pediatric population were assessed, with a higher proportion of female. In pediatric population, those with poorly controlled asthma usually had severe or persistent disease. In elderly, an increased asthma severity was found, although proper treatment might be effective. Most studies (70%) also described exacerbations, hospitalizations (48%), quality of life (39%), and emergency visits (30%). Despite heterogeneity of outcomes and population, studies show an important prevalence of uncontrolled asthma even in patients being treated, with better disease control with treatment improvements. CONCLUSIONS: Studies in Brazil have shown that asthma control remains a challenge and there is still a need for improvement on disease management.

Enhancing the expression of multi-antigen chimeric TGAGS/BST protein from Toxoplasma gondii in Escherichia coli BL 21 Star during batch cultivation.

Toxoplasmosis, despite advances in science and technology, is a disease that requires attention since there is no vaccine capable of immunizing humans and animals against all isolated types of Toxoplasma gondii. Thus, the use of chimeric proteins, which can contain multiple antigens, is a very promising alternative for the process of obtaining a vaccine and diagnostic test for toxoplasmosis due to the great diversity of antigens presented by T. gondii. In this context, the present study evaluates batch culture strategies in the production of the multi-antigenic chimeric protein TgAGS/BsT from Toxoplasma gondii. Several exploratory cultures were initially carried out to observe the kinetic behavior of E. coli BL21 Star in five different medium compositions without the addition of IPTG (inducer). Cultures of E. coli B21 Star were carried out with 1.0 mM IPTG at different times of initiation of induction (0.5, 1, and 6 h) to evaluate the effects on cell growth, production of the protein of interest, culture pH, and acetic acid formation. The results showed that among the culture media evaluated, 2xTY and TB supplemented with glycerol had the best cell concentration values of 3.42 ± 0.05 g/L and 5.48 ± 0.05 g/L, respectively. In the assays induced by IPTG, a higher expression of TgAGS/BsT protein was observed, with induction beginning within 6 h of culture, with a maximum concentration of protein of interest of 1.82 ± 0.02 g/L for the 2xTY and 2.49 ± 0.03 g/L for the TB medium. In addition, later induction by IPTG provided greater stability of plasmid pET-TgAGS, remaining with values above 90% at the end of culture.