Dual-Target Radiosurgery for Concomitant Continuous Pain Presentation of Trigeminal Neuralgia: Radiomodulation Effect and Dose.

OBJECTIVES: Patients with trigeminal neuralgia (TN) experience concomitant continuous pain (CCP) that can be difficult to treat. A dual-target approach delivering a high dose of radiation to the nerve and the contralateral thalamus can develop a fast radiomodulation effect on lowering pain. We sought to determine if this effect was dose dependent. METHODS: We retrospectively reviewed 21 patients treated with radiosurgery in CCP and severe TN pain, with a visual analog scale (VAS) score of nine out of 10 at the time of treatment. Ten patients were treated with a high dose (>120 Gy) in the thalamus 90 Gy to the nerve, and the rest with a low dose (<120 Gy) to the thalamus and >90 Gy to the nerve. RESULTS: Of those who received the high dose to the thalamus, six patients (60%) received 140 Gy, and four (40%) received 120 Gy, with a median dose to the trigeminal nerve of 90 and 85 Gy, respectively. The high thalamus dose showed a radiomodulation effect from day 1. The low thalamus dose did not produce radiomodulation on any of the first four days. The percentage of VAS score reduction one month after treatment was higher in the high-thalamus dose group than in the low-thalamus dose group. At three months, VAS score was 2 in the high-dose group and 4 in the low-dose group. CONCLUSIONS: The radiomodulation effect in pain and dual-target radiosurgery is dose dependent in CCP in TN; a high dose can provide a more consistent clinical result than a lower dose.

Tissue-specific overexpression of systemic RNA interference components limits lifespan in C. elegans.

Intertissue RNA transport recently emerged as a novel signaling mechanism. In mammals, mounting evidence suggests that small RNA transfer between cells is widespread and used in various physiological contexts. In the nematode C. elegans, a similar mechanism is conferred by the systemic RNAi pathway. Members of the Systemic RNA Interference Defective (SID) family act at different steps of cellular RNA uptake and export. The limiting step in systemic RNA interference (RNAi) is the import of extracellular RNAs via the conserved double-stranded (dsRNA)-gated dsRNA channel SID-1. To better understand the role of RNAs as intertissue signaling molecules, we modified the function of SID-1 in specific tissues of C. elegans. We observed that sid-1 loss-of-function mutants are as healthy as wild-type worms. Conversely, overexpression of sid-1 in C. elegans intestine, muscle, or neurons rendered worms short-lived. The effects of intestinal sid-1 overexpression were attenuated by silencing the components of systemic RNAi sid-1, sid-2 and sid-5, implicating systemic RNA signaling in the lifespan reduction. Accordingly, tissue-specific overexpression of sid-2 and sid-5 also reduced worm lifespan. Additionally, an RNAi screen for components of several non-coding RNA pathways revealed that silencing the miRNA biogenesis proteins PASH-1 and DCR-1 rendered the lifespan of worms with intestinal sid-1 overexpression similar to controls. Collectively, our data support the notion that systemic RNA signaling must be tightly regulated, and unbalancing that process provokes a reduction in lifespan. We termed this phenomenon Intercellular/Extracellular Systemic RNA imbalance (InExS).

Depression as a risk factor for adverse outcomes and increased healthcare utilization in bariatric surgery patients.

INTRODUCTION: Depression is strongly associated with obesity and is common among patients undergoing bariatric surgery. Little is known about the impact of depression on early postoperative outcomes or its association with substance use. METHODS: The Michigan Bariatric Surgery Collaborative is a statewide quality improvement program that maintains a large clinical registry. We evaluated patients undergoing primary Roux-en-Y gastric bypass or sleeve gastrectomy between 2017 and 2022. Patients self-reported symptoms of depression (PHQ-8) and use of alcohol (AUDIT-C), smoking, prescription opiates, and marijuana at baseline. Preoperative PHQ-8 scores stratified patients based on severity: no depression (0-4), mild (5-9), moderate (10-14), or severe (15-24). We compared 30-day outcomes and substance use between patients with and without depression. RESULTS: Among 44,301 patients, 30.8% had some level of depression, with 19.8% mild, 7.5% moderate, and 3.5% severe. Patients with depression were more likely to have an extended length of stay (LOS) (> 3 days) than those without depression (no depression 2.1% vs. severe depression 3.0%, p = 0.0452). There were no significant differences between no depression and severe depression groups in rates of complications (5.7% vs. 5.2%, p = 0.1564), reoperations (0.9%, vs. 0.8%, p = 0.7394), ED visits (7.7% vs. 7.8%, p = 0.5353), or readmissions (3.2% vs. 3.9%, p = 0.3034). Patients with severe depression had significantly higher rates of smoking (9.7% vs. 12.5%, p < 0.0001), alcohol use disorder (8.6% vs. 14.0%, p < 0.0001), opiate use (14.5% vs. 22.4%, p < 0.0001) and marijuana use (8.4%, vs. 15.5%, p = 0.0008). CONCLUSIONS: This study demonstrated that nearly one-third of patients undergoing bariatric surgery have depression, with over 10% in the moderate to severe range. There was a significant association between preoperative depressive symptoms and extended LOS after bariatric surgery, as well as higher rates of smoking and use of marijuana, prescription opiates and alcohol. There was no significant effect on adverse events or other measures of healthcare utilization.

Upfront Radiosurgery for Treatment of Symptomatic Obstructive Hydrocephalus due to Brain Tumors.

Introduction Hydrocephalus is a build-up of cerebrospinal fluid (CSF) in the brain and is characterized by abnormal dilatation of the cerebral ventricles. Patients can be either asymptomatic, have symptoms related to primary tumors, or have hydrocephalus-related symptoms. Generally, symptomatic patients are candidates for ventriculoperitoneal (VP) shunt placement to reduce acute symptoms. Little evidence exists regarding the resolution of symptomatic hydrocephalus secondary to brain tumors using stereotactic radiosurgery (SRS) alone as a primary treatment option. Methods The present study is a retrospective series of eight patients (six men and two women) diagnosed with obstructive hydrocephalus due to brain tumors treated with radiosurgery between April 2013 and February 2021. The primary endpoint of the present study is to report our institutional experience regarding the control of symptomatic obstructive hydrocephalus due to brain tumors treated with upfront radiosurgery. Results The mean age was 52 years (range, 5-79). The most common presenting symptoms included headache (100%), vision-related symptoms (75%), and ataxia (37.5%). All patients showed symptom improvement after radiosurgery, five (62.5%) patients showed resolution in less than three days and the rest of the patients resolved hydrocephalus in a longer timeframe (more than three days). All patients lowered their Evans index compared to the index documented before radiosurgery, in a range from 0.02 to 0.17. Conclusion Radiosurgery is a non-invasive alternative treatment for primary and secondary brain tumors that debut with obstructive hydrocephalus, tumors expected to have a high alpha/beta ratio might be suitable to attempt radiosurgery to avoid permanently implanted devices such as VP shunts or other invasive procedures such as a third ventriculostomy. The present study demonstrated that in selected cases SRS can lead to hydrocephalus symptom resolution along with a decrease in ventricular size in a relatively short time frame. Little evidence exists regarding the effect of SRS on symptomatic hydrocephalus resolution and further histology-specific studies are required. We acknowledge that this approach requires immediate access to radiosurgery and close clinical follow-up to ensure success.

Visual Symptoms Outcomes in Cavernous Sinus Radiosurgery and a Systematic Review.

Introduction The complex anatomy of the cavernous sinus confers a true challenge when it comes to safe tumor resection. Due to its non-invasive nature, stereotactic radiosurgery (SRS) is expected to have lower mortality and morbidity rates than microsurgery. The purpose of this study was to evaluate clinical results regarding visual symptoms after SRS for benign tumors invading the cavernous sinus. We also conducted a systematic literature review to provide a robust analysis regarding visual outcomes. Methods The study included 58 patients (43 women and 15 men; mean age: 52 years) with benign tumors invading the cavernous sinus (27 pituitary adenomas and 31 meningiomas) who underwent SRS with different platforms between August 2011 and December 2021. Of these, 26 patients underwent surgery before SRS, and the remaining 32 had SRS as first-line therapy. We identified symptoms involving cranial nerves (CN) II, III, IV, and VI in 38 patients at the time of SRS. We conducted a systematic review to identify all original studies assessing visual outcomes. We searched PubMed, the Latin American and Caribbean Health Sciences Literature index, and Google Scholar using the Medical Subject Heading search terms "radiosurgery" and "cavernous sinus" for valid studies published until January 31, 2022. Results Regarding pituitary adenomas, median tumor volume was 2.05 cc, 3.12 cc, and 2.39 cc for Gamma Knife (GK), CyberKnife (CK), and tomotherapy (Tomo), respectively. Median doses were 14 Gy for GK, 17 Gy for CK, and 15 Gy for Tomo. For meningiomas, median tumor volume was 10.2 cc, 2.62 cc, and 16.3 cc for GK, CK, and Tomo, respectively. The median dose was 14 Gy for GK, 14 Gy for CK, and 14.5 Gy for Tomo. The overall tumor control rate was 100% with a median follow-up of 33 months (range: 6-128 months). A reduction of >30% in total tumor size per the Response Evaluation Criteria in Solid Tumors (RECIST) classification was documented in seven patients (RECIST II; 12.1%), 51 patients (87.9%) had stable disease (RECIST III), and no increase in tumor volume was documented in any patient. Visual symptoms improved in 51.7% of patients. In the systematic review, the mean visual improvement was 36% (range: 25.8-42.5%). Conclusion SRS is an effective treatment for benign tumors invading the cavernous sinus. In this series, patients who underwent SRS as a primary treatment showed improvement in pre-existing cranial neuropathy and visual symptoms. Given the natural history of these tumors, which tend to grow and cause visual alternations, treating asymptomatic patients is a feasible approach worth considering for the appropriate patients.

Predictors of poor outcomes of respiratory syncytial virus acute lower respiratory infections in children under 5 years of age in a middle-income tropical country based on the National Public Health Surveillance System.

OBJECTIVES: The aim of the present study is to gain insight into the identification of region-specific factors associated with poor outcomes in children under 5 years of age with confirmed respiratory syncytial virus acute lower respiratory infection (RSV-ALRI) living in Colombia, a middle-income country, based on the National Public Health Surveillance System of the country. METHODS: An analytical cross-sectional study was conducted using epidemiological data from the records of morbidity and mortality of respiratory infections as registered in the surveillance system report of the National Institute of Health of Colombia 2018, including children under 5 years of age with confirmed RSV-ALRI. Predictor variables included demographic and clinical variables, as well as variables measured after hospital attendance. Outcome variables analyzed were respiratory failure, the need for pediatric intensive care unit admission, and mortality. RESULTS: Of a total of 8470 patients with a diagnosis of ALRI, we selected 1215 (14.3%) that were under 5 years of age and were positive for RSV. After controlling for potential confounders, it was found that age, gender, socioeconomic stratum, incomplete pneumococcal conjugate vaccine 13 immunization for age, cardiac disease, and malnutrition as comorbidities, chest X-ray findings, and development of sepsis independently predicted poor outcomes among the patients analyzed. CONCLUSIONS: The identified predictors for poor outcomes in RSV-affected children may be helpful for guiding efficient and targeted national and/or regional programs and public policies to assist in achieving Goal 3 of the Sustainable Development Goals adopted by the United Nations in 2015.

Equilibrio ocupacional durante el confinamiento por Covid-19 en docentes de terapia ocupacional de una institución universitaria / Equilíbrio ocupacional durante o confinamento por Covid-19 em professores de terapia ocupacional de uma instituição universitária / Occupational balance during Covid-19 lockdown among occupational therapy academics

Resumen Introducción El confinamiento obligatorio en respuesta al Covid-19 obligó al cierre de institutos y universidades, haciendo que los docentes adaptarán sus estrategias de enseñanza a medios digitales, experimentando un incremento de la jornada laboral, mayor estrés, fatiga y dificultades para separar el trabajo de otras ocupaciones, lo cual compromete su salud física y mental. Objetivos Describir el equilibrio ocupacional de docentes de Terapia ocupacional de una institución universitaria de Cali-Colombia, durante el confinamiento por Covid-19, y establecer la relación con variables sociodemográficas y la presencia de sintomatología de ansiedad y depresión. Metodo Estudio observacional de corte transversal con 32 docentes de terapia ocupacional. La información se recopiló mediante un cuestionario sociodemográfico, los cuestionarios OBQ-E, PHQ-9 y la escala GAD-7. Se realizaron tablas de frecuencias y porcentajes para variables cualitativas y las cuantitativas se presentaron en promedio y desviación estándar. El análisis bivariado se hizo mediante análisis de correspondencia múltiple, t-student, ANOVA y correlaciones de Pearson. Resultados El 75,0% de participantes eran mujeres. El promedio en el equilibrio ocupacional fue 36,5; el 31,3% manifestó síntomas de ansiedad y 43,8% de depresión. Conclusiones La mayoría de docentes eran mujeres, sexo que se relacionó con el cuidado de adultos mayores. Las puntuaciones en equilibrio ocupacional y la presencia de síntomas de ansiedad y depresión reflejan los ajustes que los docentes asumieron durante el confinamiento, en respuesta al aumento de las actividades diarias, la poca variedad entre ocupaciones que querían y que debían hacer y la satisfacción con el tiempo de descanso y sueño.

Resumo Introdução No setor de educação, o confinamento obrigatório em resposta ao Covid-19 obrigou ao fechamento de institutos e universidades, fazendo que os professores adaptassem suas estratégias de ensino às mídias digitais, vivenciando aumento da jornada de trabalho, maior estresse, cansaço e dificuldades na separação do trabalho de outras ocupações, o que compromete sua saúde física e mental. Objetivo Descrever o equilíbrio ocupacional de professores de terapia ocupacional de uma instituição universitária em Cali-Colômbia, durante o confinamento, e estabelecer a relação com variáveis ​​sociodemográficas e a presença de sintomas de ansiedade e depressão. Método Foi realizado um estudo observacional transversal com 32 professores de terapia ocupacional. As informações foram coletadas por meio de formulários Google que continham um questionário sociodemográfico, os questionários OBQ-E, PHQ-9 e a escala GAD-7. Foram elaboradas tabelas de frequências absolutas e percentuais para as variáveis ​​qualitativas, e as variáveis ​​quantitativas foram apresentadas em média e desvio padrão. A análise bivariada foi feita por meio de testes t-student, ANOVA e correlações de Pearson. Resultados 75% das participantes eram mulheres. No saldo ocupacional, a média foi de 36,5; 31,3% manifestaram sintomas de ansiedade e 43,8% de depressão. Conclusões A maioria dos professores era do sexo feminino, sexo que estava relacionado ao cuidado de idosos. As pontuações no equilíbrio ocupacional e a presença de sintomas de ansiedade e depressão refletem os ajustes que os professores fizeram durante o confinamento, em resposta ao aumento das atividades diárias, a pouca variedade entre as ocupações que desejavam e as que tinham que fazer e a satisfação com o tempo de descanso e sono.

Abstract Introduction The mandatory confinement in response to Covid-19 forced the closure of colleges and universities, leading professors to adapt their teaching strategies to digital media, These changes in teaching processes showed an increase in the workday, greater stress, fatigue, and difficulties in separating work from other occupations, which may compromise the physical and mental health of teachers. Objective To describe the occupational balance of occupational therapy teachers at a university in Cali-Colombia, during lockdown due to Covid-19, and to establish the relationship between sociodemographic variables and the presence of symptoms of anxiety and depression. Method A cross-sectional study was conducted with 32 occupational therapy professors. The information was collected using Google forms containing a sociodemographic questionnaire, the OBQ-E, PHQ-9 questionnaires, and the GAD-7 scale. Absolute frequency tables and percentages were made for qualitative variables, and quantitative variables were presented as mean and standard deviation. Bivariate analysis was performed using t-student tests, ANOVA, and Pearson's correlations. Results 75.0% of participants were women. In occupational balance the average was 36.5; meanwhile, 31.3% manifested symptoms of anxiety and 43.8% of depression. Conclusions Most of the professors were women, gender that was related to the care of older adults. The scores in occupational balance and the presence of anxiety and depression symptoms reflect the adjustments that the professors assumed during the confinement, in response to the increase in daily activities, the little variety between occupations that they wanted and had to do, and satisfaction with the job, rest and sleep time.

Estrategias lúdicas para mejorar la higiene bucal en una población con síndrome de Down / Playful strategies to improve oral health hygiene in a population with down síndrome / Estratégias lúdicas para melhorar a higiene bucal em um povoado com síndrome de Down

Resumen Objetivos: el objetivo de la investigación fue implementar estrategias lúdicas para mejorar la higiene oral de la población con Síndrome de Down en la Fundación CINDES de la ciudad de Pereira en el periodo 2017-2. Métodos: la investigación adoptó un enfoque cualitativo analizando el mundo social de acuerdo con su propia realidad y experiencia de las 12 personas diagnosticadas con SD y sus padres o cuidadores observándolos en su entorno natural y analizando sus habilidades previas acerca de las técnicas de higiene oral. El método de investigación fue la investigación acción y se usaron estrategias lúdicas, la observación y diarios de campo como técnicas de investigación. Resultados: se evidenció en la mayoría de la población con SD y sus padres o cuidadores que el nivel de conocimiento inicial era regular por el tiempo insuficiente durante el cepillado, falta de uso de la seda dental y mal comportamiento durante el cepillado; con la aplicación de estrategias lúdicas se logró que las personas con síndrome de Down aprendieran a identificar las estructuras de la cavidad oral, los implementos de higiene oral y los riesgos de no realizar un cuidado bucal adecuado, además, mejoraron la ejecución de la técnica de cepillado y se generó cierta autonomía. Conclusiones: se logró mostrar la efectividad de la implementación de estrategias lúdicas como herramienta crucial para lograr un aprendizaje significativo.

Abstract Objectives: The objective of this research was to implement playful strategies to improve the oral hygiene of the population with Down Syndrome (DS) in the CINDES Foundation in the city of Pereira during the 2017-2 period. Method: The research adopted a qualitative approach analyzing the social world according to the own reality and experience of 12 people diagnosed with DS and their parents or caregivers, observing them in their natural environment and analyzing their previous skills regarding oral hygiene techniques. The method applied was action research and playful strategies. Observation and field diaries were used as research techniques. Results: In the majority of the population with DS and their parents or caregivers, it was evidenced that their initial level of knowledge was mediocre due to insufficient time during brushing, lack of use of dental floss and bad behavior during brushing. With the application of playful strategies, it was possible for people with Down Syndrome to learn to identify the structures of the oral cavity, of oral hygiene implements and the risks of not performing adequate oral care. In addition, they improved the execution of the brushing technique and some autonomy was generated. Conclusions: It was possible to show the effectiveness of the implementation of playful strategies as an crucial tool to achieve meaningful learning.

Resumo Objetivos: o objetivo da pesquisa foi realizar estratégias lúdicas para melhorar a higiene oral num grupo com Síndrome de Down na Fundação CINDES da cidade de Pereira no período 2017-2. Métodos: a pesquisa adoptou um enfoque qualitativo analisando o mundo social de acordo com sua própria realidade e experiência das 12 pessoas diagnosticadas com SD e seus pais ou cuidadores observando-os em seu ambiente natural e analisando suas habilidades previas acerca das técnicas de higiene oral. O método de pesquisa foi a pesquisa ação e se usaram estratégias lúdicas, a observação e diários de campo como técnicas de pesquisa. Resultados: evidenciou-se na maioria deste grupo com SD e seus pais ou cuidadores que no nível de conhecimento inicial era regular pelo tempo insuficiente durante o escovado, falta de uso do fio dental e mal comportamento durante a escovação; com a aplicação de estratégias lúdicas se conseguiu que as pessoas com síndrome de Down aprenderam a identificar as estruturas da cavidade oral, o kit de higiene oral e os riscos de não realizar um cuidado bucal adequado, além disso melhora a execução da técnica de escovação e se gerou certa autonomia. Conclusões: conseguiu-se amostrar a efetividade da implementação de estratégias lúdicas como ferramenta crucial para lograr uma aprendizagem significativa.

Stereotactic Radiosurgery for Recurrent Glioblastoma Multiforme: A Retrospective Multi-Institutional Experience.

Introduction Glioblastoma multiforme (GBM) is the most common and lethal primary malignancy of the central nervous system. Despite standard therapy protocols, such as aggressive surgical resection, radiotherapy, and chemotherapy, GBM's aggressive nature produces low survival rates. Tumor recurrence and progression are nearly universal. Stereotactic radiosurgery (SRS) has been studied as an alternative treatment for recurrent GBM as a minimally invasive option that might prolong survival. The objective of this retrospective study was to evaluate the efficacy of SRS as a treatment modality considering overall survival (OS) in patients with GBM who had tumor recurrence and were treated with SRS in three different institutions. Materials and methods We retrospectively reviewed patients who received SRS for recurrent GBM between 1992 and 2020. A total of 46 patients were included in this study. We recorded age at diagnosis, the extent of surgical resection, radiation treatment, chemotherapy regimen, Karnofsky Performance Status at the time of SRS and at last follow-up, use of adjuvant chemotherapy after SRS, and response evaluation criteria in solid tumors. Primary endpoints were OS after initial diagnosis and OS from the date of the SRS procedure. Results Patients received SRS at a median of 10 months (range, 1 to 94 months) after their initial diagnoses. Median follow-up was seven months from the time of SRS and 22.8 months since diagnosis. The estimated median OS for all patients was nine months (range, 1 to 42 months) after SRS and 23.8 months (range, 4 to 102 months) after diagnosis. Median OS after SRS was seven months for patients treated from 1992 to 2011 and nine months for those treated from 2012 to 2020 (p = 0.008; X2 = 7.008). Median OS for younger patients (i.e., those aged <50 years) was 37.1 months compared to 18.6 months for older patients (i.e., those aged >50 years; p = 0.04; X2 = 3.870). Patients who received SRS after 10 months since diagnosis had a median OS of 36.2 months versus those who received SRS sooner than 10 months, who had an OS of 15 months (p = 0.004; X2 = 8.145). Radiosurgery doses larger than 15 Gy correlated with a median survival of nine months versus seven months in those treated with doses <15 Gy (p = 0.01; X2 = 6.756). Lastly, patients who received adjuvant bevacizumab (BEV) and or chemotherapy after SRS had a median survival of 12 months versus seven months for patients who did not receive any additional therapy after SRS (p = 0.04; X2 = 4.196). Conclusion SRS focal recurrent GBM in selected patients may improve OS, especially when combined with adjuvant therapy such as BEV and chemotherapy. Other prognostic variables proved relevant such as patients' age, the dose delivered, and surgery-to-SRS time that translates to the time of recurrence. Our results were consistent with the published literature and added to the accumulating evidence regarding SRS in recurrent GBM; however, extensive, multi-center studies are required to make definitive recommendations on this treatment approach.

Characterisation of the interaction of guanine nucleotides with ribosomal GTPase Lsg1.

Ribosome biogenesis in eukaryotes requires the participation of several transactivation factors that are involved in the modification, assembly, transport and quality control of the ribosomal subunits. One of these factors is the Large subunit GTPase 1 (Lsg1), a protein that acts as the release factor for the export adaptor named Nonsense-mediated mRNA decay 3 protein (Nmd3) and facilitates the incorporation of the last structural protein uL16 into the 60S subunit. Here, we characterised the recombinant yeast Lsg1 and studied its catalysis and binding properties for guanine nucleotides. We described the interaction of Lsg1 with guanine nucleotides alone and in the presence of the complex Nmd3•60S using fluorescence spectroscopy. Lsg1 has a greater affinity for GTP than for GDP suggesting that in the cell cytoplasm it exists mainly bound to the former. In the presence of 60S subunits loaded with Nmd3, the affinity of Lsg1 for both nucleotides increases but to a larger extent towards GTP. From this observation together with the excess of GTP present in the cytoplasm of exponentially growing cells over that of GDP, we can infer that the pre-ribosomal particle composed by Nmd3•60S acts as a GTP Stabilising Factor for Lsg1. Additionally, Lsg1 undergoes different conformational changes depending on its binding partner or the guanine nucleotides it interacts with. Steady-state kinetic analysis of free Lsg1 indicated slow GTP hydrolysis with values of kcat 1 min-1 and Km of 34 µM.

Substitutions in Spike and Nucleocapsid proteins of SARS-CoV-2 circulating in South America.

SARS-CoV-2 is a new member of the genus Betacoronavirus, responsible for the COVID-19 pandemic. The virus crossed the species barrier and established in the human population taking advantage of the spike protein high affinity for the ACE receptor to infect the lower respiratory tract. The Nucleocapsid (N) and Spike (S) are highly immunogenic structural proteins and most commercial COVID-19 diagnostic assays target these proteins. In an unpredictable epidemic, it is essential to know about their genetic variability. The objective of this study was to describe the substitution frequency of the S and N proteins of SARS-CoV-2 in South America. A total of 504 amino acid and nucleotide sequences of the S and N proteins of SARS-CoV-2 from seven South American countries (Argentina, Brazil, Chile, Ecuador, Peru, Uruguay, and Colombia), reported as of June 3, and corresponding to samples collected between March and April 2020, were compared through substitution matrices using the Muscle algorithm. Forty-three sequences from 13 Colombian departments were obtained in this study using the Oxford Nanopore and Illumina MiSeq technologies, following the amplicon-based ARTIC network protocol. The substitutions D614G in S and R203K/G204R in N were the most frequent in South America, observed in 83% and 34% of the sequences respectively. Strikingly, genomes with the conserved position D614 were almost completely replaced by genomes with the G614 substitution between March to April 2020. A similar replacement pattern was observed with R203K/G204R although more marked in Chile, Argentina and Brazil, suggesting similar introduction history and/or control strategies of SARS-CoV-2 in these countries. It is necessary to continue with the genomic surveillance of S and N proteins during the SARS-CoV-2 pandemic as this information can be useful for developing vaccines, therapeutics and diagnostic tests.

Molecular analysis of several in-house rRT-PCR protocols for SARS-CoV-2 detection in the context of genetic variability of the virus in Colombia.

The COVID-19 pandemic caused by SARS-CoV-2 is a public health problem unprecedented in the recent history of humanity. Different in-house real-time RT-PCR (rRT-PCR) methods for SARS-CoV-2 diagnosis and the appearance of genomes with mutations in primer regions have been reported. Hence, whole-genome data from locally-circulating SARS-CoV-2 strains contribute to the knowledge of its global variability and the development and fine tuning of diagnostic protocols. To describe the genetic variability of Colombian SARS-CoV-2 genomes in hybridization regions of oligonucleotides of the main in-house methods for SARS-CoV-2 detection, RNA samples with confirmed SARS-CoV-2 molecular diagnosis were processed through next-generation sequencing. Primers/probes sequences from 13 target regions for SARS-CoV-2 detection suggested by 7 institutions and consolidated by WHO during the early stage of the pandemic were aligned with Muscle tool to assess the genetic variability potentially affecting their performance. Finally, the corresponding codon positions at the 3' end of each primer, the open reading frame inspection was identified for each gene/protein product. Complete SARS-CoV-2 genomes were obtained from 30 COVID-19 cases, representative of the current epidemiology in the country. Mismatches between at least one Colombian sequence and five oligonucleotides targeting the RdRP and N genes were observed. The 3' end of 4 primers aligned to the third codon position, showed high risk of nucleotide substitution and potential mismatches at this critical position. Genetic variability was detected in Colombian SARS-CoV-2 sequences in some of the primer/probe regions for in-house rRT-PCR diagnostic tests available at WHO COVID-19 technical guidelines; its impact on the performance and rates of false-negative results should be experimentally evaluated. The genomic surveillance of SARS-CoV-2 is highly recommended for the early identification of mutations in critical regions and to issue recommendations on specific diagnostic tests to ensure the coverage of locally-circulating genetic variants.

Genetic evolution of influenza viruses among selected countries in Latin America, 2017-2018.

OBJECTIVE: Since the 2009 influenza pandemic, Latin American (LA) countries have strengthened their influenza surveillance systems. We analyzed influenza genetic sequence data from the 2017 through 2018 Southern Hemisphere (SH) influenza season from selected LA countries, to map the availability of influenza genetic sequence data from, and to describe, the 2017 through 2018 SH influenza seasons in LA. METHODS: We analyzed influenza A/H1pdm09, A/H3, B/Victoria and B/Yamagata hemagglutinin sequences from clinical samples from 12 National Influenza Centers (NICs) in ten countries (Argentina, Brazil, Chile, Colombia, Costa Rica, Ecuador, Mexico, Paraguay, Peru and Uruguay) with a collection date from epidemiologic week (EW) 18, 2017 through EW 43, 2018. These sequences were generated by the NIC or the WHO Collaborating Center (CC) at the U.S Centers for Disease Control and Prevention, uploaded to the Global Initiative on Sharing All Influenza Data (GISAID) platform, and used for phylogenetic reconstruction. FINDINGS: Influenza hemagglutinin sequences from the participating countries (A/H1pdm09 n = 326, A/H3 n = 636, B n = 433) were highly concordant with the genetic groups of the influenza vaccine-recommended viruses for influenza A/H1pdm09 and influenza B. For influenza A/H3, the concordance was variable. CONCLUSIONS: Considering the constant evolution of influenza viruses, high-quality surveillance data-specifically genetic sequence data, are important to allow public health decision makers to make informed decisions about prevention and control strategies, such as influenza vaccine composition. Countries that conduct influenza genetic sequencing for surveillance in LA should continue to work with the WHO CCs to produce high-quality genetic sequence data and upload those sequences to open-access databases.

Burden of influenza-associated respiratory hospitalizations in the Americas, 2010-2015.

BACKGROUND: Despite having influenza vaccination policies and programs, countries in the Americas underutilize seasonal influenza vaccine, in part because of insufficient evidence about severe influenza burden. We aimed to estimate the annual burden of influenza-associated respiratory hospitalizations in the Americas. METHODS: Thirty-five countries in the Americas with national influenza surveillance were invited to provide monthly laboratory data and hospital discharges for respiratory illness (International Classification of Diseases 10th edition J codes 0-99) during 2010-2015. In three age-strata (<5, 5-64, and ≥65 years), we estimated the influenza-associated hospitalizations rate by multiplying the monthly number of respiratory hospitalizations by the monthly proportion of influenza-positive samples and dividing by the census population. We used random effects meta-analyses to pool age-group specific rates and extrapolated to countries that did not contribute data, using pooled rates stratified by age group and country characteristics found to be associated with rates. RESULTS: Sixteen of 35 countries (46%) contributed primary data to the analyses, representing 79% of the America's population. The average pooled rate of influenza-associated respiratory hospitalization was 90/100,000 population (95% confidence interval 61-132) among children aged <5 years, 21/100,000 population (13-32) among persons aged 5-64 years, and 141/100,000 population (95-211) among persons aged ≥65 years. We estimated the average annual number of influenza-associated respiratory hospitalizations in the Americas to be 772,000 (95% credible interval 716,000-829,000). CONCLUSIONS: Influenza-associated respiratory hospitalizations impose a heavy burden on health systems in the Americas. Countries in the Americas should use this information to justify investments in seasonal influenza vaccination-especially among young children and the elderly.

LDL Cholesterol Uptake Assay Using Live Cell Imaging Analysis with Cell Health Monitoring.

The regulation of LDL cholesterol uptake through LDLR-mediated endocytosis is an important area of study in various major pathologies including metabolic disorder, cardiovascular disease, and kidney disease. Currently, there is no available method to assess LDL uptake while simultaneously monitoring for health of the cells. The current study presents a protocol, using a live cell imaging analysis system, to acquire serial measurements of LDL influx with concurrent monitoring for cell health. This novel technique is tested in three human cell lines (hepatic, renal tubular epithelial, and coronary artery endothelial cells) over a four-hour time course. Moreover, the sensitivity of this technique is validated with well-known LDL uptake inhibitors, Dynasore and recombinant PCSK9 protein, as well as by an LDL uptake promoter, Simvastatin. Taken together, this method provides a medium-to-high throughput platform for simultaneously screening pharmacological activity as well as monitoring of cell morphology, hence cytotoxicity of compounds regulating LDL influx. The analysis can be used with different imaging systems and analytical software.

Tumor pardo en esqueleto axial de dos pacientes con enfermedad renal crónica en diálisis / Brown tumor in the axial skeleton of two patients with chronic kidney disease in dialysis

Resumen Los tumores pardos son una forma localizada de osteítis fibrosa no neoplásica, secundaria a hiperparatiroidismo (primario o secundario). Hacen parte de las alteraciones del metabolismo mineral y óseo de la enfermedad renal crónica (ERC). Se manifiestan como lesiones líticas, expansivas, asociadas a masas de tejidos blandos, que pueden estar localizadas en cualquier parte del esqueleto, con predilección por las costillas, clavículas, pelvis, fémur, huesos faciales y mandíbula. Reportamos dos casos de paciente con ERC en terapia de reemplazo renal (TRR), con hiperparatiroidismo secundario y tumores pardos localizados en mandíbula, arcos costales y cuerpos vertebrales, con manifestaciones radiológicas atípicas. Conclusión: los tumores pardos hacen parte de las alteraciones óseas de los pacientes con ERC. El aspecto benigno en los estudios de imágenes (lesiones expansivas sin destrucción de la cortical) en el contexto de un paciente con hiperparatiroidismo, pueden sugerir el diagnóstico. (Acta Med Colomb 2018; 43: 221-225).

Abstract Brown tumors are a localized form of non-neoplastic osteitis fibrosa, secondary to hyperparathyroidism (primary or secondary). They are part of the alterations of the mineral and bone metabolism of chronic kidney disease (CKD). They manifest as lytic, expansive lesions associated to soft tissue masses that can be located in any part of the skeleton with predilection for the ribs, clavicles, pelvis, femur, facial bones and jaw. Two cases of patients with CKD in renal replacement therapy (RRT), with secondary hyperparathyroidism and brown tumors located in the jaw, costal arches and vertebral bodies, with atypical radiological manifestations are described. Conclusion: brown tumors are part of the bone disorders of patients with CKD. The benign appearance in imaging studies (expansive lesions without destruction of the cortex) in the context of a patient with hyperparathyroidism, may suggest the diagnosis. (Acta Med Colomb 2018; 43: 221-225).

Micro-RNA Profiling of Exosomes from Marrow-Derived Mesenchymal Stromal Cells in Patients with Acute Myeloid Leukemia: Implications in Leukemogenesis.

Gene regulatory networks in AML may be influenced by microRNAs (miRs) contained in exosomes derived from bone marrow mesenchymal stromal cells (MSCs). We sequenced miRs from exosomes isolated from marrow-derived MSCs from patients with AML (n = 3) and from healthy controls (n = 3; not age-matched). Known targets of mIRs that were significantly different in AML-derived MSC exosomes compared to controls were identified. Of the five candidate miRs identified by differential packaging in exosomes, only miR-26a-5p and miR-101-3p were significantly increased in AML-derived samples while miR-23b-5p, miR-339-3p and miR-425-5p were significantly decreased. Validation of the predicted change in gene expression of the potential targets was investigated by interrogating gene expression levels from public datasets of marrow-derived CD34-selected cells from patients with AML (n = 69) and healthy donors (n = 40). Two molecules with decreased gene expression in AML (EZH2 and GSK3ß) were predicted by the miR profiling and have been previously implicated in AML while three molecules were increased in AML-derived cells and have not been previously associated with leukemogenesis (KRBA2, RRBP1 and HIST2H 2BE). In summary, profiling miRs in exosomes from AML-derived MSCs allowed us to identify candidate miRs with potential relevance in AML that could yield new insights regarding leukemogenesis or new treatment strategies.

Use of Statins to Augment Progenitor Cell Function in Preclinical and Clinical Studies of Regenerative Therapy: a Systematic Review.

BACKGROUND: Mesenchymal stromal cells (MSCs) and endothelial progenitor cells (EPCs) are used in cell-based regenerative therapy. HMG CoA reductase inhibitors (statins) appear promising in blocking apoptosis, prolonging progenitor cell survival and improving their capacity to repair organ function. METHODS: We performed a systematic review of preclinical and clinical studies to clarify whether statins can improve cell-based repair of organ injury. MEDLINE, EMBASE, and PUBMED databases were searched (1947 to June 25, 2013). Controlled clinical and pre-clinical studies were included that evaluated statin therapy used alone or in combination with MSCs or EPCs in patients or animals with organ injury. RESULTS: After screening 771 citations, 100 records underwent full eligibility screening of which 38 studies met eligibility and were included in the review: Studies were grouped into pre-clinical studies that involved statin treatment in combination with cell therapy (18 studies), preclinical studies of statin therapy alone (13 studies) and clinical studies of statin therapy (7 studies). Studies addressed cardiac injury (14 studies), vascular disorders (15 studies), neurologic conditions (8 studies) and bone fractures (1 study). Pre-clinical studies of statins in combination with MSC infusion (15 studies) or EPC therapy (3 studies) were described and despite marked heterogeneity in reporting outcomes of cellular analysis and organ function, all of these cell-based pre-clinical studies reported improved organ recovery with the addition of statin therapy. Moreover, 13 pre-clinical studies involved the administration of a statin drug alone to animals. An increase in EPC number and/or function (no studies of MSCs) was reported in 11 of these studies (85 %) and improved organ function in 12 studies (92 %). We also identified 7 clinical studies and none involved the administration of cells but described an increased number and/or function of EPCs (no studies of MSCs) and improved organ function with statin therapy (1.2-fold to 35-fold improvement over controls) in all 7 studies. CONCLUSION: Our systematic review provides a foundation of encouraging results that support further study of statins in regenerative therapy to augment the number and/or function of MSCs used in cell-based repair and to augment the number and function of EPCs in vivo to repair damaged tissues. Larger studies are needed to ensure safety and confirm clinical benefits.

The effect of plasma osmolality and baroreceptor loading status on postexercise heat loss responses.

We examined the separate and combined effects of plasma osmolality and baroreceptor loading status on postexercise heat loss responses. Nine young males completed a 45-min treadmill exercise protocol at 58 ± 2% V̇o2 peak, followed by a 60-min recovery. On separate days, participants received 0.9% NaCl (ISO), 3.0% NaCl (HYP), or no infusion (natural recovery) throughout exercise. In two additional sessions (no infusion), lower-body negative (LBNP) or positive (LBPP) pressure was applied throughout the final 45 min of recovery. Local sweat rate (LSR; ventilated capsule: chest, forearm, upper back, forehead) and skin blood flow (SkBF; laser-Doppler flowmetry: forearm, upper back) were continuously measured. During HYP, upper back LSR was attenuated from end-exercise to 10 min of recovery by ∼0.35 ± 0.10 mg·min(-1)·cm(-2) and during the last 20 min of recovery by ∼0.13 ± 0.03 mg·min(-1)·cm(-2), while chest LSR was lower by 0.18 ± 0.06 mg·min(-1)·cm(-2) at 50 min of recovery compared with natural recovery (all P < 0.05). Forearm and forehead LSRs were not affected by plasma hyperosmolality during HYP (all P > 0.28), which suggests regional differences in the osmotic modulation of postexercise LSR. Furthermore, LBPP application attenuated LSR by ∼0.07-0.28 mg·min(-1)·cm(-2) during the last 30 min of recovery at all sites except the forehead compared with natural recovery (all P < 0.05). Relative to natural recovery, forearm and upper back SkBF were elevated during LBPP, ISO, and HYP by ∼6-10% by the end of recovery (all P < 0.05). We conclude that 1) hyperosmolality attenuates postexercise sweating heterogeneously among skin regions, and 2) baroreceptor loading modulates postexercise SkBF independently of changes in plasma osmolality without regional differences.

Timing of influenza epidemics and vaccines in the American tropics, 2002-2008, 2011-2014.

BACKGROUND: Influenza-associated illness results in increased morbidity and mortality in the Americas. These effects can be mitigated with an appropriately chosen and timed influenza vaccination campaign. To provide guidance in choosing the most suitable vaccine formulation and timing of administration, it is necessary to understand the timing of influenza seasonal epidemics. OBJECTIVES: Our main objective was to determine whether influenza occurs in seasonal patterns in the American tropics and when these patterns occurred. METHODS: Publicly available, monthly seasonal influenza data from the Pan American Health Organization and WHO, from countries in the American tropics, were obtained during 2002-2008 and 2011-2014 (excluding unseasonal pandemic activity during 2009-2010). For each country, we calculated the monthly proportion of samples that tested positive for influenza. We applied the monthly proportion data to a logistic regression model for each country. RESULTS: We analyzed 2002-2008 and 2011-2014 influenza surveillance data from the American tropics and identified 13 (81%) of 16 countries with influenza epidemics that, on average, started during May and lasted 4 months. CONCLUSIONS: The majority of countries in the American tropics have seasonal epidemics that start in May. Officials in these countries should consider the impact of vaccinating persons during April with the Southern Hemisphere formulation.