Targeting DNA2 overcomes metabolic reprogramming in multiple myeloma.

DNA damage resistance is a major barrier to effective DNA-damaging therapy in multiple myeloma (MM). To discover mechanisms through which MM cells overcome DNA damage, we investigate how MM cells become resistant to antisense oligonucleotide (ASO) therapy targeting Interleukin enhancer binding factor 2 (ILF2), a DNA damage regulator that is overexpressed in 70% of MM patients whose disease has progressed after standard therapies have failed. Here, we show that MM cells undergo adaptive metabolic rewiring to restore energy balance and promote survival in response to DNA damage activation. Using a CRISPR/Cas9 screening strategy, we identify the mitochondrial DNA repair protein DNA2, whose loss of function suppresses MM cells' ability to overcome ILF2 ASO-induced DNA damage, as being essential to counteracting oxidative DNA damage. Our study reveals a mechanism of vulnerability of MM cells that have an increased demand for mitochondrial metabolism upon DNA damage activation.

Transcriptional Heterogeneity Overcomes Super-Enhancer Disrupting Drug Combinations in Multiple Myeloma.

Multiple myeloma (MM) is a malignancy that is often driven by MYC and that is sustained by IRF4, which are upregulated by super-enhancers. IKZF1 and IKZF3 bind to super-enhancers and can be degraded using immunomodulatory imide drugs (IMiD). Successful IMiD responses downregulate MYC and IRF4; however, this fails in IMiD-resistant cells. MYC and IRF4 downregulation can also be achieved in IMiD-resistant tumors using inhibitors of BET and EP300 transcriptional coactivator proteins; however, in vivo these drugs have a narrow therapeutic window. By combining IMiDs with EP300 inhibition, we demonstrate greater downregulation of MYC and IRF4, synergistic killing of myeloma in vitro and in vivo, and an increased therapeutic window. Interestingly, this potent combination failed where MYC and IRF4 expression was maintained by high levels of the AP-1 factor BATF. Our results identify an effective drug combination and a previously unrecognized mechanism of IMiD resistance. SIGNIFICANCE: These results highlight the dependence of MM on IKZF1-bound super-enhancers, which can be effectively targeted by a potent therapeutic combination pairing IMiD-mediated degradation of IKZF1 and IKZF3 with EP300 inhibition. They also identify AP-1 factors as an unrecognized mechanism of IMiD resistance in MM. See related article by Neri, Barwick, et al., p. 56. See related commentary by Yun and Cleveland, p. 5. This article is featured in Selected Articles from This Issue, p. 4.

Inorganic polyphosphate's role in energy production and mitochondrial permeability transition pore opening in tick mitochondria.

Inorganic polyphosphate (polyP) is a biopolymer composed of phosphoanhydride-linked orthophosphate molecules. PolyP is engaged in a variety of cellular functions, including mitochondrial metabolism. Here, we examined the effects of polyP on electron transport chain enzymes and F1 Fo ATP synthase in tick embryos during embryonic development. The study found that polyPs containing medium and long chains (polyP15 and polyP65 ) enhanced the activity of complex I, complex II, complex III, and F1 Fo ATP synthase, while short polyP chains (polyP3 ) had no effect. The study also examined the activity of exopolyphosphatases (PPX) in various energy-demand situations. PPX activity was stimulated when ADP concentrations are high, characterizing a low-energy context. When complexes I-III and F1 Fo ATP synthase inhibitors were added in energized mitochondria, PPX activity decreased, whereas the mitochondrial uncoupler FCCP had no impact on PPX activity. Additionally, the study investigated the effect of polyP on mitochondrial swelling, finding that polyP causes mitochondrial swelling by increasing calcium effects on the mitochondrial permeability transition pore. The findings presented here to increase our understanding of the function of polyP in mitochondrial metabolism and its relationship to mitochondrial permeability transition pore opening in an arthropod model.

Targeting DNA2 Overcomes Metabolic Reprogramming in Multiple Myeloma.

DNA damage resistance is a major barrier to effective DNA-damaging therapy in multiple myeloma (MM). To discover novel mechanisms through which MM cells overcome DNA damage, we investigated how MM cells become resistant to antisense oligonucleotide (ASO) therapy targeting ILF2, a DNA damage regulator that is overexpressed in 70% of MM patients whose disease has progressed after standard therapies have failed. Here, we show that MM cells undergo an adaptive metabolic rewiring and rely on oxidative phosphorylation to restore energy balance and promote survival in response to DNA damage activation. Using a CRISPR/Cas9 screening strategy, we identified the mitochondrial DNA repair protein DNA2, whose loss of function suppresses MM cells' ability to overcome ILF2 ASO-induced DNA damage, as being essential to counteracting oxidative DNA damage and maintaining mitochondrial respiration. Our study revealed a novel vulnerability of MM cells that have an increased demand for mitochondrial metabolism upon DNA damage activation. STATEMENT OF SIGNIFICANCE: Metabolic reprogramming is a mechanism through which cancer cells maintain survival and become resistant to DNA-damaging therapy. Here, we show that targeting DNA2 is synthetically lethal in myeloma cells that undergo metabolic adaptation and rely on oxidative phosphorylation to maintain survival after DNA damage activation.

Integrated analysis of next generation sequencing minimal residual disease (MRD) and PET scan in transplant eligible myeloma patients.

Minimal residual disease (MRD) assays allow response assessment in patients with multiple myeloma (MM), and negativity is associated with improved survival outcomes. The role of highly sensitive next generation sequencing (NGS) MRD in combination with functional imaging remains to be validated. We performed a retrospective analysis on MM patients who underwent frontline autologous stem cell transplant (ASCT). Patients were evaluated at day 100 post-ASCT with NGS-MRD and positron emission tomography (PET-CT). Patients with ≥ 2 MRD measurements were included in a secondary analysis for sequential measurements. 186 patients were included. At day 100, 45 (24.2%) patients achieved MRD negativity at a sensitivity threshold of 10-6. MRD negativity was the most predictive factor for longer time to next treatment (TTNT). Negativity rates did not differ according to MM subtype, R-ISS Stage nor cytogenetic risk. PET-CT and MRD had poor agreement, with high rates of PET-CT negativity in MRD-positive patients. Patients with sustained MRD negativity had longer TTNT, regardless of baseline risk characteristics. Our results show that the ability to measure deeper and sustainable responses distinguishes patients with better outcomes. Achieving MRD negativity was the strongest prognostic marker and could help guide therapy-related decisions and serve as a response marker for clinical trials.

Incidence and risk factors of pressure injuries in critically ill patients with COVID-19 / Incidencia y factores de riesgo de lesiones por presión en pacientes críticos con COVID-19 / Incidência e fatores de risco de lesão por pressão em pacientes críticos com COVID-19

ABSTRACT Objective: to analyze pressure injury (PI) incidence and risk factors in patients with COVID-19 admitted to an Intensive Care Unit and characterize the identified PIs. Method: a retrospective cohort study, consisting of 668 patients, carried out between March 2020 and February 2021. Clinical/demographic and PI variables were collected from medical records and electronic database. Data were analyzed using descriptive and inferential statistics. Logistic regression was performed to analyze risk factors for PI. Results: PI incidence was 30.2% (n=202), with the majority located in the sacral region (52.9%) and in stage 1 (39%). Risk factors were age (p<0.001), Diabetes Mellitus (p=0.005), length of stay (p<0.001), immunosuppression (p=0.034), nutritional risk (p=0.015) and mechanical ventilation (p<0.001). Conclusion: PI incidence in critically ill patients with COVID-19 was high.

RESUMEN Objetivo: analizar la incidencia y factores de riesgo de lesiones por presión (LP) en pacientes con COVID-19 ingresados en Unidad de Cuidados Intensivos y caracterizar las LP identificadas. Método: estudio de cohorte retrospectivo, compuesto por 668 pacientes, realizado entre marzo de 2020 y febrero de 2021. Las variables clínico/demográficas y de LP se recogieron de historias clínicas y bases de datos electrónicas. Los datos fueron analizados mediante estadística descriptiva e inferencial. Se realizó una regresión logística para analizar los factores de riesgo de LP. Resultados: la incidencia de LP fue del 30,2% (n=202), localizándose la mayoría en la región sacra (52,9%) y en estadio 1 (39%). Los factores de riesgo fueron edad (p<0,001), Diabetes Mellitus (p=0,005), tiempo de estancia hospitalaria (p<0,001), inmunosupresión (p=0,034), riesgo nutricional (p=0,015) y ventilación mecánica (p<0,001). Conclusión: la incidencia de LP en pacientes críticos con COVID-19 fue alta.

RESUMO Objetivo: analisar a incidência e os fatores de risco de lesão por pressão (LP) em pacientes com COVID-19 internados em Unidade de Terapia Intensiva e caracterizar as LP identificadas. Método: estudo de coorte retrospectivo, constituído por 668 pacientes, realizado entre março de 2020 e fevereiro de 2021. Variáveis clínicas/demográficas e das LP foram coletadas dos prontuários e banco de dados eletrônico. Os dados foram analisados por meio de estatística descritiva e inferencial. A regressão logística foi realizada para análise dos fatores de risco para LP. Resultados: a incidência de LP foi de 30,2% (n=202), sendo a maioria localizada na região sacral (52,9%) e em estágio 1 (39%). Os fatores de risco foram idade (p<0,001), Diabetes Mellitus (p=0,005), tempo de internação (p<0,001), imunossupressão (p=0,034), risco nutricional (p=0,015) e ventilação mecânica (p<0,001). Conclusão: a incidência de LP em pacientes críticos com COVID-19 foi alta.

Obstetric and neonatal adverse events and association with care models: a cohort study / Eventos adversos obstétricos y neonatales y asociación con modelos de atención: un estudio de cohorte / Eventos adversos obstétricos e neonatais e associação com os modelos de assistência: um estudo coorte

ABSTRACT Objective: to analyze the prevalence of practices in childbirth care, obstetric and neonatal adverse events and their association with care models in three public health services. Method: this is a prospective cohort study carried out with 548 dyads, postpartum women and their newborns, whose pregnancy was of usual risk and vaginal birth, admitted to hospital in three public services, one with an exclusive care model by nurse-midwives (service A), one with a collaborative model involving obstetric doctors and nurses (service B) and one with an exclusive medical care model (service C). Initially, an interview was carried out with participants, and a second contact was carried out 42 days after birth to complement the collection of the adverse event outcome. Results: in service A, no woman underwent the Kristeller maneuver, episiotomy, directed pushing or more than one vaginal examination per hour. Meanwhile, in service C, 19.3%, 39.9%, 77.1% and 26.3% of women underwent these interventions, respectively. Adverse events occurred in 19.2% of the dyads. Occurrence of adverse events was associated with not using partograph (p=0.001; OR: 11.03; CI: 2.64-45.99) and episiotomy (p=0.042; OR: 1.72; CI: 1. 02-2.91). The mean probability of experiencing an adverse event was 5% in service A, 21% in service B and 24% in service C. Conclusion: adverse events had a lower mean probability of occurrence in the service exclusively operated by nurse-midwives, in which greater application of recommendations for labor and birth care was identified.

RESUMEN Objetivo: analizar la prevalencia de prácticas en la atención del childbirth, eventos adversos obstétricos y neonatales y su asociación con los modelos de atención en tres servicios públicos de salud. Método: estudio de cohorte prospectivo realizado con 548 binomios, puérperas y sus recién nacidos, cuyo embarazo fue de riesgo habitual y childbirth vaginal, hospitalizados en tres servicios públicos, uno con modelo de atención exclusiva por enfermeras obstétricas (servicio A), otro con modelo colaborativo. modelo con la labor de médicos y enfermeras obstetras (servicio B) y otro con un modelo de asistencia médica exclusiva (servicio C). Inicialmente se realizó una entrevista a los participantes y se realizó un segundo contacto a los 42 días del nacimiento para complementar la recopilación del resultado del evento adverso. Resultados: en el servicio A ninguna mujer fue sometida a maniobra de Kristeller, episiotomía, pujo dirigido o más de un examen vaginal por hora. Mientras tanto, en el servicio C, el 19,3%, 39,9%, 77,1% y 26,3% de las mujeres se sometieron a estas intervenciones, respectivamente. Los eventos adversos ocurrieron en el 19,2% de los binomios. La aparición de eventos adversos se asoció con la no utilización del partographa (p=0,001; OR: 11,03; IC: 2,64-45,99) y episiotomía (p=0,042; OR: 1,72; IC: 1,02-2,91). La probabilidad promedio de experimentar un evento adverso fue del 5% en el servicio A, del 21% en el servicio B y del 24% en el servicio C. Conclusión: los eventos adversos tuvieron menor probabilidad promedio de ocurrencia en el servicio operado exclusivamente por enfermeras obstétricas, en el que se identificó mayor aplicación de las recomendaciones de asistencia al childbirth y nacimiento.

RESUMO Objetivo: analisar a prevalência de práticas na atenção ao parto, eventos adversos obstétricos e neonatais e sua associação com modelos assistenciais em três serviços de saúde públicos. Método: estudo coorte prospectivo realizado com 548 binômios, puérperas e seus recém-nascidos, cuja gestação foi de risco habitual e o parto vaginal, internados em três serviços públicos, sendo um com modelo de assistência exclusivo por enfermeiras obstetras (serviço A), um com modelo colaborativo com atuação de médicos e enfermeiras obstetras (serviço B) e um com modelo de assistência exclusiva médica (serviço C). Inicialmente, foi realizada uma entrevista com as participantes e um segundo contato foi realizado após 42 dias do parto para complementar a coleta do desfecho evento adverso. Resultados: no serviço A, nenhuma mulher foi submetida à manobra de Kristeller, episiotomia, incentivos a puxos dirigidos ou mais de um toque vaginal por hora. Enquanto, no serviço C, 19,3%, 39,9%, 77,1% e 26,3% das mulheres foram submetidas a essas intervenções, respectivamente. Os eventos adversos ocorreram em 19,2% dos binômios. A ocorrência dos eventos adversos foi associada ao não uso do partograma (p=0,001; OR: 11,03; IC: 2,64-45,99) e episiotomia (p=0,042; OR: 1,72; IC: 1,02-2,91). A probabilidade média de apresentar algum evento adverso foi de 5% no serviço A, 21% no serviço B e 24% no serviço C. Conclusão: os eventos adversos apresentaram menor probabilidade média de ocorrência no serviço com atuação exclusiva de enfermeiras obstetras, no qual se identificou maior aplicação das recomendações para assistência ao parto e nascimento.

Time trend of audiological procedures in the Brazilian Public Health System / Tendência temporal de procedimentos audiológicos no Sistema Único de Saúde

ABSTRACT Purpose: to analyze the time trend of audiological procedures in the Brazilian Public Health System (SUS) between 2008 and 2019 in the geographical regions of Brazil. Methods: a time series ecological study with data on audiological procedures from the SUS Outpatient Information System. The study calculated annual indices of audiological procedures with joinpoint regression for the trend analysis. The progress in the provision of procedures between the initial and final years in the series and the distribution of identified records was calculated. Results: altogether, 38,500,404 records were found in the study period. The South (178.84/10,000 inhabitants) and North of Brazil (130.97/10,000 inhabitants), respectively, had the highest and lowest mean procedure indices. Both were the only regions with a trend towards a significant increase in procedures. In Brazil, they increased by 56.91%, with an emphasis on the increase in "otoacoustic evoked emissions in neonatal hearing screening" and a decrease in "visual reinforcement audiometry (air/bone conduction)." In the distribution, procedures concentrated on basic audiological assessments. Conclusion: audiological procedures have increased at SUS, but there are discrepancies between procedures and geographical regions of Brazil.

RESUMO Objetivo: analisar a tendência temporal de procedimentos audiológicos no Sistema Único de Saúde (SUS), entre 2008 e 2019, nas regiões geográficas brasileiras. Métodos: estudo ecológico de série temporal, com dados de procedimentos audiológicos do Sistema de Informações Ambulatoriais do SUS. Foram calculadas taxas anuais de procedimentos audiológicos, utilizando-se a regressão Joinpoint para análise de tendência. Calculou-se a evolução da oferta destes procedimentos entre os anos inicial e final da série e a distribuição dos registros identificados. Resultados: foram localizados 38.500.404 registros no período investigado. Verificou-se que as regiões Sul (178,84/10.000 habitantes) e Norte (130,97/10.000 habitantes) apresentaram a maior e a menor taxa média de procedimentos, respectivamente. Ambas foram as únicas regiões que exibiram tendências de aumento significativo na realização dos procedimentos. No Brasil, a oferta dos procedimentos demonstrou uma evolução positiva de 56,91%, com destaque para crescimento de "emissões otoacústicas evocadas para triagem auditiva (teste da orelhinha)" e queda de "audiometria de reforço visual (via aérea/óssea)". Na distribuição, houve concentração de procedimentos da avaliação audiológica básica. Conclusão: a realização de procedimentos audiológicos no SUS revela aumento, mas evidencia discrepâncias nas regiões geográficas brasileiras e nos procedimentos executados.

Cross-cultural differences in beliefs about emotions: A comparison between Brazilian and British participants / Diferenças transculturais em crenças sobre emoções: uma comparação entre participantes brasileiros e britânicos

ABSTRACT Objective: Perfectionist beliefs about emotions impact the experience and expression of emotions, being linked to increased levels of depression and anxiety. Given the influence of culture in the representation and expression of emotion, it is possible that beliefs vary across countries, but few empirical studies have been conducted on the theme. This study aims to compare Brazilian and British samples regarding their beliefs about emotional experience and expression. Methods: The current study compared a total of 960 Brazilian and British participants, with the samples having a similar profile in terms of age, gender and ethnicity. Participants answered online the Generalized Anxiety Disorder (GAD-7), Patient Health Questionnaire (PHQ-9), and Beliefs about Emotions Scale (BES). This study aims to compare Brazilian and British samples with regard to beliefs about emotional experience and expression. Results: Significant differences between samples were found for beliefs about emotions. As hypothesized, Brazilians scored lower on unhelpful beliefs about emotions, except for beliefs about experiencing negative feelings and emotional control. Differences in total BES scores remained even after the inclusion of depression and anxiety as covariates. Conclusions: Results suggest higher endorsement of perfectionist beliefs in a European versus a Latin American context, but highlight that this pattern depends on the specific beliefs being studied. These differences should be considered when working with people from different cultural backgrounds and developing cultural adaptations for clinical interventions and psychopathology models.

RESUMO Objetivo: Crenças perfeccionistas sobre emoções afetam a experiência e expressão de emoções, estando relacionadas a níveis aumentados de depressão e ansiedade. Dada a influência da cultura na representação e expressão de emoções, é possível que as crenças variem entre os países, mas poucos estudos empíricos foram realizados sobre o tema. Este estudo tem o objetivo de comparar amostras brasileiras e britânicas em relação às suas crenças sobre a experiência emocional e expressão. Métodos: O presente estudo comparou um total de 960 participantes brasileiros e britânicos, com as amostras tendo um perfil semelhante em termos de idade, gênero e etnia. Os participantes responderam on-line à Escala de Ansiedade Generalizada (GAD-7), ao Questionário de Saúde do Paciente (PHQ-9) e à Escala de Crenças sobre Emoções (BES). Resultados: Foram encontradas diferenças significativas entre as amostras em relação às crenças sobre emoções. Como hipotetizado, os brasileiros obtiveram pontuações mais baixas em crenças prejudiciais sobre emoções, exceto nas crenças sobre experienciar sentimentos negativos e controle emocional. As diferenças nos escores totais da BES permaneceram mesmo após a inclusão de depressão e ansiedade como covariáveis. Conclusões: Os resultados sugerem maior endosso de crenças perfeccionistas em um contexto europeu, em comparação com um contexto latino-americano, mas destacam que esse padrão depende das crenças específicas estudadas. Essas diferenças devem ser consideradas ao trabalhar com pessoas de diferentes origens culturais e no desenvolvimento de adaptações culturais para intervenções clínicas e modelos de psicopatologia.

SARS-CoV-2 Spatiotemporal Genomic and Molecular Analysis of the First Wave of the COVID-19 Pandemic in Macaé, the Brazilian Capital of Oil.

The SARS-CoV-2 virus infection led to millions of deaths during the COVID-19 pandemic. Hundreds of workers from several other Brazilian cities, as well as from other countries, arrive daily in Macaé to work in the oil supply chain, making this city a putative hotspot for the introduction of new viral lineages. In this study, we performed a genomic survey of SARS-CoV-2 samples from Macaé during the first outbreak of COVID-19, combined with clinical data and a molecular integrative analysis. First, phylogenomic analyses showed a high occurrence of viral introduction events and the establishment of local transmissions in Macaé, including the ingression and spread of the B.1.1.28 lineage in the municipality from June to August 2020. Second, SARS-CoV-2 mutations were identified in patients with distinct levels of COVID-19 severity. Third, molecular interactions of the mutated spike protein from three B.1.1.33 local samples and human ACE2 showed higher interactions than that of the wild-type spike protein from the ancestral virus. Altogether, these results elucidate the SARS-CoV-2 genomic profile in a strategic Brazilian city and further explore the functional aspects of SARS-CoV-2 with a characterization of emerging viral mutations associated with clinical data and the potential targets for drug development against SARS-CoV-2.

Educación médica y bienestar: orientaciones para el desarrollo de una educación médica positiva / Medical education and well-being: orientations towards the implementation of a positive teaching

The transition to and through college can affect the mental health of college students. The situation in medical students is particularly worrisome, considering that they have high rates of psychopathology. These mental health problems can be associated with worse academic and professional outcomes and worse patient care. In this scenario, it is relevant to look at how this problem can be addressed considering variables of both mental health and educational systems in medicine. Therefore, initiatives to promote well-being during the career are proposed. The importance of well-being at an institutional level should be defined. Also, the foundations of learning neurosciences from the point of view of well-being should be incorporated. Considering that well-being goes beyond the absence of discomfort, the role of educational environment both in learning and in mental health should be recognized. Proven effective interventions should be incorporated. It must be considered that not every initiative linked to well-being is necessarily positive and that discomfort to a certain extent can be exploited educatively as well. The well-being of medical students is a complex, dynamic and multidimensional issue. These principles are orientations for different actors of the educational process, to define how they will approach this problem in their contexts.

Molecular testing and analysis of disease spreading during the emergence of COVID-19 in Macaé, the Brazilian National Capital of Oil.

The Brazilian strategy to overcome the spread of COVID-19 has been particularly criticized due to the lack of a national coordinating effort and an appropriate testing program. Here, a successful approach to control the spread of COVID-19 transmission is described by the engagement of public (university and governance) and private sectors (hospitals and oil companies) in Macaé, state of Rio de Janeiro, Brazil, a city known as the National Oil Capital. In 2020 between the 17th and 38th epidemiological week, over two percent of the 206,728 citizens were subjected to symptom analysis and RT-qPCR testing by the Federal University of Rio de Janeiro, with positive individuals being notified up to 48 h after swab collection. Geocodification and spatial cluster analysis were used to limit COVID-19 spreading in Macaé. Within the first semester after the outbreak of COVID-19 in Brazil, Macaé recorded 1.8% of fatalities associated with COVID-19 up to the 38th epidemiological week, which was at least five times lower than the state capital (10.6%). Overall, considering the successful experience of this joint effort of private and public engagement in Macaé, our data suggest that the development of a similar strategy countrywise could have contributed to a better control of the COVID-19 spread in Brazil. Quarantine decree by the local administration, comprehensive molecular testing coupled to scientific analysis of COVID-19 spreading, prevented the catastrophic consequences of the pandemic as seen in other populous cities within the state of Rio de Janeiro and elsewhere in Brazil.

Can roughage: concentrate ratio affect the action of red propolis extract on sheep metabolism?

The aim of the present study was to evaluate whether the addition of red propolis extract (RPE), in different roughage: concentrate (R:C) ratios, influences the intake, digestibility, ruminal parameters, and serum biochemistry of sheep. We used eight Santa Inês sheep with an average body weight of 29.45 ± 1.58 kg, housed in metabolism cages for 60 days, and distributed in two simultaneous Latin square designs in a 2 × 2 factorial scheme. The factors consisted of two R:C ratios (70:30 and 30:70) with or without the addition of 15 mL/day of RPE. No interactions were found (P > 0.05) between R:C ratios and with or without RPE. Sheep fed 30:70 ratio showed higher (P < 0.05) intake and dry matter (DM) digestibility and non-fibrous carbohydrates and lower (P < 0.05) intake and neutral detergent fiber (NDF) digestibility. The addition of RPE did not influence (P > 0.05) the intake or nutrients digestibility, but the sheep that received RPE had a higher (P < 0.05) ruminal pH and longer (P < 0.05) time of rumination (min/kg DM) compared to the group without propolis. The ruminal ammonia concentration was higher for sheep fed 70:30 ratio, but the concentrations of total protein and albumin did not differ between R:C ratios. The addition of 15 mL of RPE does not influence the intake, digestibility, ingestive behavior, and rumen ammonical nitrogen of sheep. There is no association between the R:C ratio and the addition of 15 mL/day of RPE for sheep.

Understanding small ORF diversity through a comprehensive transcription feature classification.

Small open reading frames (small ORFs/sORFs/smORFs) are potentially coding sequences smaller than 100 codons that have historically been considered junk DNA by gene prediction software and in annotation screening; however, the advent of next-generation sequencing has contributed to the deeper investigation of junk DNA regions and their transcription products, resulting in the emergence of smORFs as a new focus of interest in systems biology. Several smORF peptides were recently reported in non-canonical mRNAs as new players in numerous biological contexts; however, their relevance is still overlooked in coding potential analysis. Hence, this review proposes a smORF classification based on transcriptional features, discussing the most promising approaches to investigate smORFs based on their different characteristics. First, smORFs were divided into non-expressed (intergenic) and expressed (genic) smORFs. Second, genic smORFs were classified as smORFs located in non-coding RNAs (ncRNAs) or canonical mRNAs. Finally, smORFs in ncRNAs were further subdivided into sequences located in small or long RNAs, whereas smORFs located in canonical mRNAs were subdivided into several specific classes depending on their localization along the gene. We hope that this review provides new insights into large-scale annotations and reinforces the role of smORFs as essential components of a hidden coding DNA world.

Establishment of suitable reference genes for studying relative gene expression during the transition from trophozoites to cyst-like stages and first evidences of stress-induced expression of meiotic genes in Trichomonas vaginalis.

Trichomonas vaginalis is a parasite of the human urogenital tract and the causative agent of trichomoniasis, a sexually transmitted disease of worldwide importance. This parasite is usually found as a motile flagellated trophozoite. However, when subjected to stressful microenvironmental conditions, T. vaginalis trophozoites can differentiate into peculiar cyst-like stages, which exhibit notable physiological resistance to unfavourable conditions. Although well documented in morphological and proteomic terms, patterns of gene expression changes involved in the cellular differentiation into cyst-like stages are mostly unknown. The real-time reverse transcription polymerase chain reaction (RT-qPCR) is recognized as a sensitive and accurate method for quantification of gene expression, providing fluorescence-based data that are proportional to the amount of a target RNA. However, the reliability of relative expression studies depends on the validation of suitable reference genes, which RNAs exhibit a minimum of variation between tested conditions. Here, we attempt to determine suitable reference genes to be used as controls of invariant expression during cold-induced in vitro differentiation of T. vaginalis trophozoites into cyst-like forms. Furthermore, we reveal that the mRNA from the meiotic recombinase Dmc1 is upregulated during this process, indicating that cryptic sexual events may take place in cyst-like stages of T. vaginalis.

Análise temporal da concessão de aparelhos de amplificação sonora individual pelo Sistema Único de Saúde / Temporal analysis of hearing aids provision by the Brazilian Unified Health System

RESUMO Objetivo Analisar a produção ambulatorial de aspectos da concessão de aparelhos de amplificação sonora individual (AASI) pelo Sistema Único de Saúde (SUS) entre 2005 e 2018. Método Trata-se de um estudo ecológico, de série temporal, baseado em dados do Sistema de Informações Ambulatoriais do SUS e do Instituto Brasileiro de Geografia e Estatística. As taxas de concessão de AASI foram analisadas com o software Joinpoint. Realizaram-se análises descritivas para categorias tecnológicas, financiamento e reposição de AASI e acompanhamento audiológico. Resultados As regiões Sul (9,96/10.000 habitantes) e Norte (3,20/10.000 habitantes) registraram a maior e menor média das taxas, respectivamente. Houve tendências de aumento significativas da concessão de AASI no Brasil e nas regiões Norte, Sudeste e Centro-Oeste, com posteriores tendências de redução significativas nas regiões Sudeste e Centro-Oeste. No país, a concessão das categorias tecnológicas foi de A (39,26%), B (36,93%) e C (23,81%), elevando o financiamento. As regiões Centro-Oeste (24,78%) e Nordeste (14,22%) apresentaram a maior e menor proporção da reposição de AASI, respectivamente. O acompanhamento audiológico predominou na região Sudeste (45,88%), com menor ocorrência na região Norte (4,18%). Conclusão Entre 2005 e 2018, há oscilações de tendências e discrepâncias entre as regiões geográficas quanto à concessão de AASI pelo SUS, além de descompassos no fornecimento das categorias tecnológicas e financiamento, considerável proporção de reposições e insuficiente quantitativo de acompanhamento audiológico para usuários de AASI.

ABSTRACT Purpose To analyze the outpatient production of aspects of hearing aids (HA) provision by the Brazilian Unified Health System (SUS) between 2005 and 2018. Methods This is an ecological time-series study, based on data from the Outpatient Information System of the SUS and the Brazilian Institute of Geography and Statistics. The HA provision rates were analyzed with Joinpoint software. Descriptive analyses were carried out for technological categories, funding, HA replacement and audiological monitoring. Results The South (9.96/10,000 inhabitants) and North (3.20/10,000 inhabitants) regions recorded the highest and lowest average rates, respectively. There were significant upward trends in HA provision in Brazil, in the North, Southeast and Midwest regions, with subsequent significant downward trends in the Southeast and Midwest. In the country, HA concession by technological categories was A (39.26%), B (36.93%) and C (23.81%), increasing funding. The Midwest (24.78%) and Northeast (14.22%) regions had the highest and lowest proportion of HA replacement, respectively. The audiological monitoring predominated in the Southeast (45.88%), with the lowest occurrence in the North (4.18%). Conclusion Between 2005 and 2018, trends fluctuated and discrepancies between geographic regions were observed in HA provision by the SUS, in addition to mismatches in the provision of technological categories and funding, a considerable proportion of replacements and insufficient audiological monitoring for HA users.

Femoral fractures in the elderly in Brasil - incidence, lethality, and costs (2008-2018)

SUMMARY OBJECTIVES: To describe the incidence by gender and region, lethality, and costs associated with the treatment of femoral fractures in the elderly (≥ 60 years) hospitalized in the Unified Health System (SUS) of Brasil between 2008 and 2018. METHODS: This is a cross-sectional, descriptive, retrospective study of hospitalizations of elderly people due to femoral fractures by analyzing secondary data obtained from the SUS Hospital Information System (SIH/SUS) between 2008 and 2018; for calculation of epidemiological coefficients, we used information from demographic censuses (2000 and 2010) of the Brazilian Geography and Statistics Institute (IBGE). RESULTS: A total of 478,274 hospitalizations were recorded in the period; the incidence was 1.7 times higher in females (overall average of 274.91/100,000 for women and 161/100,000 for men). The Southeast region had the highest absolute number of hospitalizations and the South region presented the highest annual overall average incidence (224.02/100,000). The average annual cost for SUS for the treatment of femoral fractures in the elderly was R$ 99,718,574.30. CONCLUSIONS: In the evaluated period (2008-2018), femoral fractures in the elderly had a high incidence (478,274 hospitalizations; 224.02 cases/100,000 elderly), a predominance of females (1.7F/1.0M), a higher absolute number of hospitalizations in the Southeast region and a higher incidence in the South region; the lethality was high (an increase of 17.46%; overall mean coefficient of 4.99%/year); and the costs for the SUS were huge (an increase of 126.24%; average annual expenditure of R$ 99,718,574.30).

RESUMO OBJETIVOS: Descrever a incidência por gênero e região, a letalidade e os custos associados ao tratamento de fraturas do fêmur em idosos (≥ 60 anos) internados no Sistema Único de Saúde (SUS) do Brasil entre 2008 e 2018. MÉTODOS: Estudo transversal, descritivo e retrospectivo das internações de idosos por fraturas do fêmur mediante análise dos dados secundários obtidos do Sistema de Informações Hospitalares do SUS (SIH/SUS) entre 2008 e 2018; para cálculo dos coeficientes epidemiológicos, utilizamos informações dos censos demográficos (2000 e 2010) do Instituto Brasileiro de Geografia e Estatística (IBGE). RESULTADOS: Foram registradas 478.274 mil internações no período. A incidência foi 1,7 vezes maior no gênero feminino (média geral de 274,91/100.000 para mulheres e 161/100.000 para homens). A região sudeste obteve maior número absoluto de internações e na região sul apresentou a maior incidência média geral anual (224,02/100.000). O custo médio anual do SUS para o tratamento das fraturas de fêmur em idosos foi de R$ 99.718.574,30. CONCLUSÕES: No período avaliado (2008-2018), as fraturas do fêmur em idosos apresentaram alta incidência (478.274 mil internações; 224,02 casos/100.000 idosos), predomínio do gênero feminino (1,7F/1,0M), maior número absoluto de internações na região sudeste e maior incidência na região sul; a letalidade foi elevada (aumento de 17,46%; média geral do coeficiente de 4,99%/ano); e, os custos para o SUS foram vultuosos (aumento de 126,24%, média anual de gastos de R$ 99.718.574,30).

Satisfação de usuários com aparelhos de amplificação sonora individual concedidos pelo Sistema Único de Saúde: revisão integrativa / Satisfaction of users with hearing aids provided by the Unified Health System: an integrative review

RESUMO Objetivo Identificar a satisfação de usuários com os aparelhos de amplificação sonora individual (AASI) concedidos pelo Sistema Único de Saúde (SUS). Estratégia de pesquisa Trata-se de uma revisão integrativa da literatura, realizada nas bases de dados LILACS, SciELO, PubMed e Scopus, empregando os descritores hearing loss, public health policy, Unified Health System, public health, patient satisfaction e hearing aids. Critérios de seleção Foram selecionados artigos publicados a partir de 2004, sem restrição quanto ao idioma, envolvendo usuários adaptados pelo SUS. Excluíram-se publicações repetidas, resenhas, artigos de opinião, editoriais, teses e dissertações. Resultados Foram localizados 1011 estudos, dos quais, 24 foram incluídos. As pesquisas veicularam-se a partir de 2007, com predomínio na região Sudeste, por meio de abordagens quantitativas e, em grande parte, com amostras limitadas, compreendendo adultos e idosos. Os questionários de autoavaliação foram os recursos utilizados para avaliar a satisfação. Conclusão A maioria dos usuários revelou elevada satisfação com os AASI concedidos pelo SUS.

ABSTRACT Purpose Identify user satisfaction with hearing aids (HAs) provided by the Unified Health System (UHS). Research strategy This is an integrative literature review, carried out in the LILACS, SciELO, PubMed and Scopus databases, using the following keywords: "hearing loss", "public health policy", "Unified Health System", "public health", "patient satisfaction" and "hearing aids". Selection criteria Articles published from 2004 onwards, without language restrictions, involving users treated by the UHS were selected. Duplicate publications, reviews, opinion articles, editorials, theses and dissertations were excluded. Results A total of 1011 studies were found, 24 of which were included. The studies were published from 2007 onwards, with a predominance in the Southeast region, using quantitative approaches with limited samples comprising adults and older people. Self-assessment questionnaires were used to evaluate satisfaction. Conclusion Most users showed a high level of satisfaction with the HAs provided by the UHS.

Inorganic pyrophosphatase from the red flour beetle (Tribolium castaneum) modulates mitochondrial polyphosphate metabolism.

Polyphosphates (polyPs) have been found in all cell types examined to date and play diverse roles, depending on the cell type. In eukaryotic organisms, polyPs have been mainly investigated in mammalian cells, with few studies on insects. In this study, we investigated mitochondrial polyphosphate metabolism in the red flour beetle, Tribolium castaneum. Substrate specificity for different chain lengths demonstrated the presence of two exopolyphosphatase isoforms in mitochondria. T. castaneum mitochondrial polyP levels decreased after injection with soluble pyrophosphatase (Tc-sPPase) dsRNA, while the membrane exopolyphosphate activity increased. Mitochondrial respiration modulated exopolyphosphatase activity only in wild-type beetles. Tripolyphosphate was able to increase the F-ATPase activity in wild-type and Tc-sPPase RNAi beetles. We suggest that inorganic pyrophosphatase modulates polyphosphate metabolism in mitochondria and affects the link between mitochondrial activity and polyphosphate metabolism in T. castaneum.