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Gene expression through RT-qPCR can be performed by the relative quantification method, which requires the expression normalization through reference genes. Therefore, it is essential to validate, experimentally, the candidate reference genes. Thus, although there are several studies that are performed to identify the most stable reference genes, most them validate genes for very specific conditions, not exploring the whole potential of the research since not all possible combinations of treatments and/or conditions of the study are explored. For this reason, new experiments must be conducted by researchers that have interest in analyzing gene expression of treatments and/or conditions present, but not explored, in these studies. Here, we present the RGeasy tool, which aims to facilitate the selection of reference genes, allowing the user to choose genes for a greater number of combinations of treatments/conditions, compared to the ones present in the original articles, through just a few clicks. RGeasy was validated with RT-qPCR data from gene expression studies performed in two coffee species, Coffea arabica and Coffea canephora, and it can be used for any animal, plant or microorganism species. In addition to displaying a rank of the most stable reference genes for each condition or treatment, the user also has access to the primer pairs for the selected reference genes., (© 2024. The Author(s).)

Context: Studies that analyze the temporal trend and spatial clustering of medical education indicators are scarce, especially in developing countries such as Brazil. This analysis is essential to subsidize more equitable policies for the medical workforce in the states and regions of Brazil. Thus, this study aimed to analyze the temporal trend and identify spatial clusters of medical education indicators in Brazil disaggregated by public and private education, states, and regions., Methods: A time-series ecological study was conducted using data from the Higher Education Census of the Ministry of Education from 2010 to 2021. The study analyzed vacancy density indicators of active and former students/100,000 population, disaggregated by public and private education, 27 states, and 5 regions in Brazil. Prais-Winsten regression was used for trend analyses of indicators. Hot Spot Analysis (Getis-Ord Gi*) was used to identify spatial clusters of indicators., Results: The number of medical schools increased by 102.2% between 2010 and 2021. A total of 366 medical schools offered 54,870 vacancies at the end of 2021. Vacancy density and active and former students increased significantly in the period, but this increase was greater in private institutions. Most states and regions showed an increasing trend in the indicators, with higher increase percentages in private than in public schools. Hot spot spaces changed over time, concentrated in the southeast, center-west, and north at the end of 2021. Medical education remains uneven in Brazil, with a low provision in regions with low socioeconomic development, academic structure, and health services, represented by regions in the north and northeast., Conclusions: There is a growing trend in medical education indicators in Brazil, especially in the private sector. Spatial clusters were found predominantly in the southeast, center-west, and north. These results indicate the need for more equitable medical education planning between the regions., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Background: So far, three porcine hemoplasmas (PH) have been identified, namely Mycoplasma suis, Mycoplasma parvum, and Mycoplasma haemosuis. The first one is the main agent associated with porcine hemoplasmosis, a possible cause of economic losses in pig production. Thus, this work aimed to detect and quantify PH 16S rRNA in finishing pigs and to associate its load estimate with average daily weight gain (ADWG). For this purpose, whole blood samples from 318 pigs were collected at an age of 75 days (d0) when the pigs entered the finishing phase and 105 days later (d105). To calculate ADWG, the animals were weighed at the abovementioned dates. Then, DNA from blood samples were submitted to a qPCR targeting the 16S rRNA gene for PH. Spearman correlation test was performed to investigate potential associations between ADWG and the quantification values. Lastly, the molecular characterization of PH was done by sequencing the 23S rDNA gene., Results: Out of the 318 samples, 190 (59.74%) were positive on d0, and 304 (95.6%) were positive on d105. A significant correlation was observed (p < 0.05), albeit with a low coefficient value (0.18), when comparing ADWG with quantification values on d105. The phylogenetic analysis based on the 23S rDNA gene showed that four sequences were closely related to M. parvum, and one sequence was positioned in the M. suis cluster., Conclusion: Two PH, M. suis and M. parvum, were detected in a Brazilian pig farm. Moreover, increasing occurrence through time was observed, which may have affected the productive performance of positive animals, mainly at the end of the finishing phase, when antimicrobials are removed.

Background: This cross-sectional study evaluated the adequacy of the Family Health Strategy for the primary care model for chronic noncommunicable diseases and the changes that occurred between the two cycles of external evaluations of the National Program for Improving Access and Quality of Primary Care, which took place in 2012 and 2014, in the higher coverage context of the Family Health Strategy of Brazil, in the state of Tocantins, Brazil., Methods: The data source contained information on the infrastructure of the 233 Primary Health Units and on the work process of 266 health teams. The Principal Component Analysis for Categorical Data technique and the McNemar chi-squared statistical test for comparing paired samples were used, and a significance level of 5% with a 95% Confidence Interval was used., Results: The analysis identified a low proportion of dispensing of medications for the treatment of chronic disease in both cycles. There was a significant increase in seasonal influenza vaccination, in the number of sterilization, procedure, dressings and inhalation rooms. There was a small but significant reduction in the materials for cervical cancer screening, although they are available in almost 90.0% of the PHUs. More than 70.0% of the health teams carried out additional health education activities, encouraged physical activity, registered schoolchildren with health needs for monitoring, evaluated user satisfaction and user referral., Conclusions: The findings of this study highlighted the improvement of the structure of the Primary Health Units, but identified a low provision of medicines to treat chronic diseases. The health promotion was performed as the main work process tool of family health teams, but it was little focused on intersectoral actions and on actions with the population in the area or on the empowerment of users through self-management support for chronic diseases. Furthermore, it is critical that the Family Health Strategy in Tocantins be organized and focused on the care of chronic diseases to improve and adapt itself to a primary chronic care model.

Background: Both diarrhea in calves and mastitis in cows limit cattle production. The bacteria involved in these diseases have shown multi-resistance to antimicrobials, however plant metabolites therefore can provide an alternative method of control. This study selected and characterized Cerrado plant extracts showing inhibitory effects against Escherichia coli and Staphylococcus spp. from cattle. Thirteen leaf extracts were initially screened and diameters of inhibition zones produced against the pathogens were recorded using an agar disk diffusion method. Total condensed tannin contents were determined and antibacterial activities were analyzed after tannin removal from the five selected extracts. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were evaluated by macro-dilution antimicrobial susceptibility tests, and the extracts were characterized by high performance liquid chromatography., Results: Inter- and intra-specific bacterial variations in the susceptibility to the extracts were detected. The aqueous extract (AE) from Caryocar brasiliense Cambess. leaves produced larger inhibition zones against E. coli strains than did other selected extracts. However, the AE from Schinopsis brasiliensis was the most effective against Staphylococcus spp. strains (P < 0.001). The MIC of ethanolic extracts (EE) from C. brasiliense (0.27 mg/mL) and S. brasiliensis (0.17 mg/mL) were lower than those of other extracts. The MIC and MBC of the Annona crassiflora EE were 6.24 mg/mL for all bacteria. Flavonoids were the main metabolites detected in the A. crassiflora EE as well as in the AE and EE from C. brasiliense, while tannins were the main metabolites in the S. brasiliensis leaf extracts., Conclusion: The AE from C. brasiliense was more effective against Gram-negative bacteria, while the AE from S. brasiliensis was more effective against Gram-positive bacteria. A. crassiflora EE and S. brasiliensis extracts are potent bactericide. After removal of the tannins, no antimicrobial effects were observed, indicating that these metabolites are the main active antibacterial components.

Background: Oral cancer is a potentially fatal disease, especially when diagnosed in advanced stages. In Brazil, the primary health care (PHC) system is responsible for promoting oral health in order to prevent oral diseases. However, there is insufficient evidence to assess whether actions of the PHC system have some effect on the morbidity and mortality from oral cancer. The purpose of this study was to analyze the effect of PHC structure and work processes on the incidence and mortality rates of oral cancer after adjusting for contextual variables., Methods: An ecological, longitudinal and analytical study was carried out. Data were obtained from different secondary data sources, including three surveys that were nationally representative of Brazilian PHC and carried out over the course of 10 years (2002-2012). Data were aggregated at the state level at different times. Oral cancer incidence and mortality rates, standardized by age and gender, served as the dependent variables. Covariables (sociodemographic, structure of basic health units, and work process in oral health) were entered in the regression models using a hierarchical approach based on a theoretical model. Analysis of mixed effects with random intercept model was also conducted (alpha = 5%)., Results: The oral cancer incidence rate was positively association with the proportion of of adults over 60 years (β = 0.59; p = 0.010) and adult smokers (β = 0.29; p = 0.010). The oral cancer related mortality rate was positively associated with the proportion of of adults over 60 years (β = 0.24; p < 0.001) and the performance of preventative and diagnostic actions for oral cancer (β = 0.02; p = 0.002). Mortality was inversely associated with the coverage of primary care teams (β = -0.01; p < 0.006) and PHC financing (β = -0.52 -9 ; p = 0.014)., Conclusions: In Brazil, the PHC structure and work processes have been shown to help reduce the mortality rate of oral cancer, but not the incidence rate of the disease. We recommend expanding investments in PHC in order to prevent oral cancer related deaths.

Background: Caregivers are responsible for the home care of family members with mental-health disorders often experience changes in their life that can generate stress and burden. The aim of this study was to identify factors associated with the burden of caregivers of family members with mental disorders., Methods: This cross-sectional study was conducted with a non-probability sample of family caregivers, whose patients attended a community services program, the Psychosocial Care Centers, in three cities in the southwest region of Goiás State, Central Brazil. Data collection took place from June 2014 to June 2015. The participants were 281 caregivers who completed a sociodemographic questionnaire and the Zarit Burden Interview (ZBI). Bivariate analyses (t test, analysis of variance, and Pearson correlation) were performed, and variables with values of p < 0.10 and gender were included in a multiple-linear regression model. Values of p < 0.05 were considered significant., Results: The caregivers were mostly female and parents of the patients, were married, with low education, and of low income. The mean ZBI score was 27.66. The factors independently associated with caregivers' burden were depression, being over 60 years of age, receiving no help with caregiving, recent patient crisis, contact days, and having other family members needing care., Conclusions: This study identified factors that deserve the attention of community services and can guide programs, such as family psycho-education groups, which may help to minimize or prevent the effects of burden on family caregivers responsible for patients' home care.

Background: Unequal distribution of emergency care services is a critical barrier to be overcome to assure access to emergency and surgical care. Considering this context it was objective of the present work analyze geographic access barriers to emergency care services in Brazil. A secondary aim of the study is to define possible roles to be assumed by small hospitals in the Brazilian healthcare network to overcome geographic access challenges., Methods: The present work can be classified as a cross-sectional ecological study. To carry out the present study, data of all 5843 Brazilian hospitals were categorized among high complexity centers and small hospitals. The geographical access barriers were identified through the use of two-step floating catchment area method. Once concluded the previous step an evaluation using the Getis-Ord-Gi method was performed to identify spatial clusters of municipalities with limited access to high complexity centers but well covered by well-equipped small hospitals., Results: The analysis of accessibility index of high complexity centers highlighted large portions of the country with nearly zero hospital beds by inhabitant. In contrast, it was possible observe a group of 1595 municipalities with high accessibility to small hospitals, simultaneously with a low coverage of high complexity centers. Among the 1595 municipalities with good accessibility to small hospitals, 74% (1183) were covered by small hospitals with at least 60% of minimum emergency service requirements. The spatial clusters analysis aggregated 589 municipalities with high values related to minimum emergency service requirements. Small hospitals in these 589 cities could promote the equity in access to emergency services benefiting more than eight million people., Conclusions: There is a spatial disequilibrium within the country with prominent gaps in the health care network for emergency services. Taking this challenge into consideration, small hospitals could be a possible solution and foster equity in access to emergency and surgical care. However more investments in are necessary to improve small hospitals capabilities to fill this gap.

Background: Physician shortage is a global issue that concerns Brazil's authorities. The organizational structure and the environment of a medical institution can hide a low-quality life of a physician. This study examines the relationship between the hospital work environment and physicians' job satisfaction and motivation when working in a large public academic hospital., Methods: The study was restricted to one large, multispecialty Brazil's hospital. Six hundred hospital physicians were invited to participate by e-mail. A short version of the Physician Worklife Survey (PWS) was used to measure working satisfaction. Physicians were also asked for socio-demographic information, medical specialty, and the intention to continue working in the hospital., Results: Data from 141 questionnaires were included in the analyses. Forty-five physicians graduated from the hospital's university, and they did not intend to leave the hospital under any circumstance (affective bond). The motivating factor for beginning the career at the hospital and to continue working there were the connection to the medical school and the hospital status as a "prestigious academic hospital"; the physicians were more satisfied with the career than the specialty. Only 30% completely agreed with the statement "If I had to start my career over again, I would choose my current specialty," while 45% completely agreed with the statement "I am not well compensated given my training and experience." The greater point of satisfaction was the relationship with physician colleagues. They are annoyed about the amount of calls they are requested to take and about how work encroaches on their personal time. No significant differences between medical specialties were found in the analysis., Conclusions: The participants were satisfied with their profession. The fact that they remained at the hospital was related to the academic environment, the relationship with colleagues, and the high prestige in which society holds the institution. The points of dissatisfaction were inadequate remuneration and the fact that work invaded personal time. Routinely, there is a need for organizations to examine the impact of their structures, policies, and procedures on the stress and quality of life of physicians.

Background: Breast cancer is the most common cause of death from cancer in women in less developed regions. Therefore, the objective of this study was to provide data on the temporal trends in female breast cancer mortality between 1990 and 2011 and to evaluate its association with the social inequalities present in Brazil., Methods: Breast cancer mortality data and estimates for the resident population were obtained from the Brazilian National Health Service database for the 1990-2011 period. Age-standardized mortality rates were calculated (20-39, 40-49, 50-69 and ≥70 years) by direct standardization using the 1960 standard world population. Trends were modeled using joinpoint regression model and linear regression. The Social Exclusion Index and the Human Development Index were used to classify the 27 Brazilian states. Pearson's correlation was used to describe the association between the Social Exclusion Index and the Human DeveIopment and the variations in mortality rates in each state., Results: Age-standardized mortality rates in Brazil were found to be stable (annual percent change [APC] = 0.3; 95% CI: -0.1 - 0.7) between 1994 and 2011. Considering the Brazilian states, significant decreases in mortality rates were found in Rio Grande do Sul, Rio de Janeiro and São Paulo. Increases in mortality rates were most notable in the states of Maranhão (APC = 11.2; 95 %CI: 5.8 - 16.9), Piauí (APC = 9.8; 95% CI: 7.6 - 12.1) and Paraíba (APC = 9.3; 95% CI: 6.0 - 12.8). There was a statistically significant correlation between Social Exclusion Index and a change in female breast cancer mortality rates in the Brazilian states between 1990 and 2011 and between Human Development Index and mortality between 2001 and 2011., Conclusions: Female breast cancer mortality rates are stable in Brazil. Reductions in these rates were found in the more developed states, possibly reflecting better healthcare.

Background: The use of lactic acid bacteria as vehicles to delivery antigens to immunize animals is a promising issue. When genetically modified, these bacteria can induce a specific local and systemic immune response against selected pathogens. Gastric acid and bile salts tolerance, production of antagonistic substances against pathogenic microorganisms, and adhesive ability to gut epithelium are other important characteristics that make these bacteria useful for oral immunization., Results: Bacteria isolated on de Man, Rogosa and Sharpe medium (MRS) from different gastrointestinal portions of broiler chicks were evaluated for their resistance to artificial gastric acid and bile salts, production of hydrogen peroxide, and cell surface hydrophobicity. Thirty-eight isolates were first typed at species level by PCR amplification of 16S-23S rRNA intergenic spacers using universal primers that anneal within 16S and 23S genes, followed by restriction digestion analyses of PCR amplicons (PCR-ARDRA). An expression cassette was assembled onto the pCR2.1-Topo vector by cloning the promoter, leader peptide, cell wall anchor and terminator sequences derived from the laminin binding S-layer protein gene of L. crispatus strain F5.7 (lbs gene). A sequence encoding the green fluorescent protein (GFP) was inserted as reporter gene, and an erythromycin resistance gene was added as selective marker. All constructs were able to express GFP in the cloning host E. coli XL1-Blue and different Lactobacillus strains as verified by FACS and laser scanning confocal microscopy., Conclusion: Lactobacillus isolated from gastrointestinal tract of broiler chickens and selected for probiotic characteristics can be genetically modified by introducing an expression cassette into the lbs locus. The transformed bacteria expressed on its cell wall surface different fluorescent proteins used as reporters of promoter function. It is possible then that similar bacterial model expressing pathogen antigens can be used as live oral vaccines to immunize broilers against infectious diseases.