Spatial imaging of glycoRNA in single cells with ARPLA.

Little is known about the biological roles of glycosylated RNAs (glycoRNAs), a recently discovered class of glycosylated molecules, because of a lack of visualization methods. We report sialic acid aptamer and RNA in situ hybridization-mediated proximity ligation assay (ARPLA) to visualize glycoRNAs in single cells with high sensitivity and selectivity. The signal output of ARPLA occurs only when dual recognition of a glycan and an RNA triggers in situ ligation, followed by rolling circle amplification of a complementary DNA, which generates a fluorescent signal by binding fluorophore-labeled oligonucleotides. Using ARPLA, we detect spatial distributions of glycoRNAs on the cell surface and their colocalization with lipid rafts as well as the intracellular trafficking of glycoRNAs through SNARE protein-mediated secretory exocytosis. Studies in breast cell lines suggest that surface glycoRNA is inversely associated with tumor malignancy and metastasis. Investigation of the relationship between glycoRNAs and monocyte-endothelial cell interactions suggests that glycoRNAs may mediate cell-cell interactions during the immune response.

Infectious endarteritis in aortic coarctation: two spectra of an infrequent disease / Endarterite infecciosa na coarctação da aorta: dois espectros de uma doença pouco frequente

ABSTRACT Objective: To describe two different degrees of clinical commitment and results in the evolution of infectious endarteritis in patients without a previous diagnosis of aortic coarctation. Case description: Two male patients aged 13 and 9 years old were admitted. The first due to a fever for 2 months, which started after dental cleaning, and the second due to high blood pressure, both patients with asthenia and weight loss. In the first case, the transthoracic echocardiogram showed aortic coarctation, and the transesophageal echocardiogram showed the presence of vegetations in the post-coarctation area, without pseudoaneurysms, with blood culture positive for Streptococcus mitis. This patient was treated for six weeks with crystalline penicillin, resolving the infection without complications. The second case was assessed for high blood pressure with a history of fever, and was treated with antibiotics. When performing a transthoracic echocardiogram, aortic coarctation was observed with a saccular image classified as a pseudoaneurysm by angiography and tomography. Blood culture was negative, and the patient developed an episode of hematemesis whose initial etiology could not be determined. Before surgical repair, he had a second episode of copious hematemesis with hypovolemic shock and death. Comments: We need to have a high index of clinical suspicion to establish the diagnosis of aortic coarctation complicated by endarteritis and start the appropriate antibiotic treatment, always maintaining surveillance for the early detection of pseudoaneurysms.

RESUMO Objetivo: Descrever dois diferentes graus de comprometimento clínico e resultados na evolução de endarterite infecciosa em pacientes sem diagnóstico prévio de coarctação da aorta. Descrição do caso: Dois pacientes do sexo masculino com idades entre 13 e nove anos foram internados. O primeiro por febre durante dois meses, iniciada após limpeza dentária. O segundo por hipertensão arterial. Ambos com astenia e perda de peso. No primeiro caso, o ecocardiograma transtorácico mostrou coarctação da aorta e o ecocardiograma transesofágico revelou vegetações na área pós-coarctação, sem pseudoaneurismas. A hemocultura foi positiva para de Streptococcus mitis. Este paciente foi tratado por seis semanas com penicilina cristalina, resolvendo a infecção sem complicações. O segundo caso foi avaliado pela presença de hipertensão arterial, com história de febre tratada com antibióticos. Ao realizar o ecocardiograma transtorácico, observou-se coarctação da aorta com imagem sacular classificada como pseudoaneurisma pela angiografia e tomografia. A hemocultura foi negativa. O paciente desenvolveu um episódio de hematêmese, cuja etiologia inicial não pôde ser determinada. Antes da correção cirúrgica, apresentou um segundo episódio de hematêmese profusa, com choque hipovolêmico e óbito. Comentários: Devemos ter um alto índice de suspeição clínica para poder estabelecer o diagnóstico de coarctação da aorta complicada com endarterite e iniciar o tratamento antibiótico adequado. É preciso manter a vigilância para a detecção precoce de pseudoaneurismas.

Infectious endarteritis in aortic coarctation: two spectra of an infrequent disease.

OBJECTIVE: To describe two different degrees of clinical commitment and results in the evolution of infectious endarteritis in patients without a previous diagnosis of aortic coarctation. CASE DESCRIPTION: Two male patients aged 13 and 9 years old were admitted. The first due to a fever for 2 months, which started after dental cleaning, and the second due to high blood pressure, both patients with asthenia and weight loss. In the first case, the transthoracic echocardiogram showed aortic coarctation, and the transesophageal echocardiogram showed the presence of vegetations in the post-coarctation area, without pseudoaneurysms, with blood culture positive for Streptococcus mitis. This patient was treated for six weeks with crystalline penicillin, resolving the infection without complications. The second case was assessed for high blood pressure with a history of fever, and was treated with antibiotics. When performing a transthoracic echocardiogram, aortic coarctation was observed with a saccular image classified as a pseudoaneurysm by angiography and tomography. Blood culture was negative, and the patient developed an episode of hematemesis whose initial etiology could not be determined. Before surgical repair, he had a second episode of copious hematemesis with hypovolemic shock and death. COMMENTS: We need to have a high index of clinical suspicion to establish the diagnosis of aortic coarctation complicated by endarteritis and start the appropriate antibiotic treatment, always maintaining surveillance for the early detection of pseudoaneurysms.

Withdrawal from repeated nicotine vapor exposure increases somatic signs of physical dependence, anxiety-like behavior, and brain reward thresholds in adult male rats.

Nicotine vapor consumption via electronic nicotine delivery systems has increased over the last decade. While prior work has shed light on the health effects of nicotine vapor inhalation, its unique effects on the brain and behavior have not been thoroughly explored. In this study we assessed markers of withdrawal following 14 days of nicotine vapor exposure. For Experiment 1, 21 adult male rats were exposed to ambient air or 6, 12, or 24 mg/mL nicotine vapor for 14 consecutive days. Following exposure on day 14, rats were injected with the nicotinic receptor antagonist mecamylamine (3.0 mg/mL) and assessed for somatic withdrawal signs and anxiety-like behavior in the elevated plus maze. For Experiment 2, 12 adult male rats were tested for intracranial self-stimulation (ICSS) immediately following exposure to vehicle vapor (50%/50%, vegetable glycerin/propylene glycol) or 24 mg/mL nicotine vapor, for 14 consecutive days. ICSS behavior was assessed for an additional 14 days, following cessation of repeated vapor exposure. Results reveal that rats with repeated nicotine vapor exposure display an increase in behavioral indicators of withdrawal following injection of mecamylamine (precipitated withdrawal). Additionally, increases in ICSS stimulation thresholds, indicative of reduced brain reward sensitivity, persist following cessation of repeated nicotine vapor exposure (spontaneous withdrawal). These data suggest that repeated e-cigarette use leads to nicotine dependence and withdrawal that affects behavior and brain reward function. Further characterization of the health effects of nicotine vapor is necessary to improve treatment strategies for nicotine use disorder and public health policies related to novel nicotine delivery systems.

Traslado no programado de pacientes pediátricos desde una sala de internación general a una unidad de cuidados intensivos / Unplanned transfer of pediatric patients from the general ward to the intensive care unit

Introducción. Durante la internación, los pacientes pueden presentar un deterioro clínico significativo y requerir el ingreso no programado a la unidad de cuidados intensivos pediátricos (UCIP). Esto puede conllevar un aumento de la morbilidad y la mortalidad. Frecuentemente, estos eventos están precedidos por una fase de deterioro que podría pasar desapercibida. Objetivo. Determinar la frecuencia, analizar las causas, describir las características clínicas y los resultados de los traslados no programados en pacientes pediátricos hospitalizados, desde el área de internación general pediátrica (IGP) a la UCIP, y analizar las diferencias entre traslados urgentes y emergentes. Población y métodos. Estudio descriptivo prospectivo; se analizaron todos los traslados no programados desde IGP a la UCIP ocurridos entre el 1 de enero de 2014 y el 31 de diciembre 2019. Resultados. Se constataron 212 traslados no programados (21 traslados cada 1000 ingresos). El 76 % de los pacientes trasladados presentaban una comorbilidad asociada ­la más frecuente fue la patología oncológica (36 %)­ y llevaban más de 24 horas internados en IGP. Las causas más frecuentes de traslado fueron dificultad respiratoria (43 %), sepsis (20 %) y complicaciones neurológicas/neuroquirúrgicas (20 %). La tasa de mortalidad global fue del 8,96 % (19 pacientes). Conclusiones. El análisis de los traslados no programados es un elemento esencial en la evaluación de la calidad de atención y seguridad del paciente de un área, y debe constituir un indicador integrado al tablero de control. La interpretación de los traslados no programados como un evento prevenible constituye un cambio de paradigma clave.

Introduction. During hospitalization, patients may develop significant clinical deterioration and require unplanned admission to the pediatric intensive care unit (PICU). This may result in increased morbidity and mortality. These events are often preceded by a deterioration phase that may go unnoticed. Objective. To determine the frequency, analyze the causes, and describe the clinical characteristics and outcomes of unplanned transfers of hospitalized pediatric patients from the general pediatric ward (GPW) to the PICU, and analyze the differences between urgent and emergent transfers. Population and methods. Prospective, descriptive study; all unplanned transfers from the GPW to the PICU occurring between January 1st, 2014 and December 31st, 2019 were analyzed. Results. There were 212 unplanned transfers (21 transfers per 1000 admissions). An associated comorbidity was present in 76% of transferred patients ­being cancer the most frequent one (36%)­ and they had been hospitalized for more than 24 hours in the GPW. The most frequent causes of transfer were respiratory distress (43%), sepsis (20%), and neurological/neurosurgical complications (20%). The overall mortality rate was 8.96% (19 patients). Conclusions. The analysis of unplanned transfers is a critical component in the assessment of the quality of care and patient safety of an area, and should be an indicator integrated into the control panel. The interpretation of unplanned transfers as a preventable event is a key paradigm shift.

Cytotoxic CD4+ T cells eliminate senescent cells by targeting cytomegalovirus antigen.

Senescent cell accumulation has been implicated in the pathogenesis of aging-associated diseases, including cancer. The mechanism that prevents the accumulation of senescent cells in aging human organs is unclear. Here, we demonstrate that a virus-immune axis controls the senescent fibroblast accumulation in the human skin. Senescent fibroblasts increased in old skin compared with young skin. However, they did not increase with advancing age in the elderly. Increased CXCL9 and cytotoxic CD4+ T cells (CD4 CTLs) recruitment were significantly associated with reduced senescent fibroblasts in the old skin. Senescent fibroblasts expressed human leukocyte antigen class II (HLA-II) and human cytomegalovirus glycoprotein B (HCMV-gB), becoming direct CD4 CTL targets. Skin-resident CD4 CTLs eliminated HCMV-gB+ senescent fibroblasts in an HLA-II-dependent manner, and HCMV-gB activated CD4 CTLs from the human skin. Collectively, our findings demonstrate HCMV reactivation in senescent cells, which CD4 CTLs can directly eliminate through the recognition of the HCMV-gB antigen.

Validation of an immunochromatographic assay kit based on colored latex particles for the identification of the canine visceral leishmaniasis.

Visceral leishmaniasis is a zoonotic infectious disease with a severe impact on humans and animals. Infection is transmitted by phlebotomine sand flies. The dogs are main reservoir for human infection. A rapid and accurate diagnosis of canine visceral leishmaniasis is essential for an efficient surveillance program. The aim of this study was to assess the performance of a rapid immunochromatographic strip test based on functionalized colored particles and a new recombinant antigenic protein, as a visual "in situ" method for the diagnosis of canine visceral leishmaniasis. The results were evaluated using an in-house ELISA assay with the same antigen. Both tests produced concordant results and the immunochromatographic strip test showed good diagnostic sensitivity (98%) and specificity (95%). Finally, meta-analysis was used to compare the sensitivity and specificity of the here developed test with the results of commercial immunochromatographic strip tests obtained from literature.

Unplanned transfer of pediatric patients from the general ward to the intensive care unit. / Traslado no programado de pacientes pediátricos desde una sala de internación general a una unidad de cuidados intensivos.

Introduction. During hospitalization, patients may develop significant clinical deterioration and require unplanned admission to the pediatric intensive care unit (PICU). This may result in increased morbidity and mortality. These events are often preceded by a deterioration phase that may go unnoticed. Objective. To determine the frequency, analyze the causes, and describe the clinical characteristics and outcomes of unplanned transfers of hospitalized pediatric patients from the general pediatric ward (GPW) to the PICU, and analyze the differences between urgent and emergent transfers. Population and methods. Prospective, descriptive study; all unplanned transfers from the GPW to the PICU occurring between January 1st , 2014 and December 31st, 2019 were analyzed. Results. There were 212 unplanned transfers (21 transfers per 1000 admissions). An associated comorbidity was present in 76% of transferred patients -being cancer the most frequent one (36%)- and they had been hospitalized for more than 24 hours in the GPW. The most frequent causes of transfer were respiratory distress (43%), sepsis (20%), and neurological/neurosurgical complications (20%). The overall mortality rate was 8.96% (19 patients). Conclusions. The analysis of unplanned transfers is a critical component in the assessment of the quality of care and patient safety of an area, and should be an indicator integrated into the control panel. The interpretation of unplanned transfers as a preventable event is a key paradigm shift.

Introducción. Durante la internación, los pacientes pueden presentar un deterioro clínico significativo y requerir el ingreso no programado a la unidad de cuidados intensivos pediátricos (UCIP). Esto puede conllevar un aumento de la morbilidad y la mortalidad. Frecuentemente, estos eventos están precedidos por una fase de deterioro que podría pasar desapercibida. Objetivo. Determinar la frecuencia, analizar las causas, describir las características clínicas y los resultados de los traslados no programados en pacientes pediátricos hospitalizados, desde el área de internación general pediátrica (IGP) a la UCIP, y analizar las diferencias entre traslados urgentes y emergentes. Población y métodos. Estudio descriptivo prospectivo; se analizaron todos los traslados no programados desde IGP a la UCIP ocurridos entre el 1 de enero de 2014 y el 31 de diciembre 2019. Resultados. Se constataron 212 traslados no programados (21 traslados cada 1000 ingresos). El 76 % de los pacientes trasladados presentaban una comorbilidad asociada ­la más frecuente fue la patología oncológica (36 %)­ y llevaban más de 24 horas internados en IGP. Las causas más frecuentes de traslado fueron dificultad respiratoria (43 %), sepsis (20 %) y complicaciones neurológicas/neuroquirúrgicas (20 %). La tasa de mortalidad global fue del 8,96 % (19 pacientes). Conclusiones. El análisis de los traslados no programados es un elemento esencial en la evaluación de la calidad de atención y seguridad del paciente de un área, y debe constituir un indicador integrado al tablero de control. La interpretación de los traslados no programados como un evento prevenible constituye un cambio de paradigma clave.

A vicious circle in breast cancer: The interplay between inflammation, reactive oxygen species, and microRNAs.

Breast cancer (BC) is the most common cancer in women worldwide. This highly heterogeneous disease is molecularly stratified into luminal A, luminal B, HER2, triple-negative/basal-like, and normal-like subtypes. An important aspect in BC progression is the activation of inflammatory processes. The activation of CD8+/Th1, NK, and M1 tumor associated macrophages (TAMs), leads to tumor destruction. In contrast, an anti-inflammatory response mediated by CD4+/Th2 and M2 TAMs will favor tumor progression. Inflammation also stimulates the production of inflammatory mediators like reactive oxygen species (ROS). In chronic inflammation, ROS activates oxidative stress and endothelial dysfunction. In cancer, ROS plays a dual role with anti-tumorigenic and pro-tumorigenic effects in cell signaling pathways that control proliferation, survival, apoptosis, and inflammation. MicroRNAs (miRNAs), which are known to be involved in BC progression and inflammation, can be regulated by ROS. At the same time, miRNAs regulate the expression of genes modulating oxidative stress. In this review, we will discuss the interplay between inflammation, ROS, and miRNAs as anticancer and tumor promoter molecules in BC. A clear understanding of the role of miRNAs in the regulation of ROS production and inflammation, may lead to new opportunities for therapy in BC.

Comparison of clinical characteristics and outcomes of hospitalized patients with seasonal coronavirus infection and COVID-19: a retrospective cohort study.

BACKGROUND: Unlike SARS-CoV and MERS-C0V, SARS-CoV-2 has the potential to become a recurrent seasonal infection; hence, it is essential to compare the clinical spectrum of COVID-19 to the existent endemic coronaviruses. We conducted a retrospective cohort study of hospitalized patients with seasonal coronavirus (sCoV) infection and COVID-19 to compare their clinical characteristics and outcomes. METHODS: A total of 190 patients hospitalized with any documented respiratory tract infection and a positive respiratory viral panel for sCoV from January 1, 2011, to March 31, 2020, were included. Those patients were compared with 190 hospitalized adult patients with molecularly confirmed symptomatic COVID-19 admitted from March 1, 2020, to May 25, 2020. RESULTS: Among 190 patients with sCoV infection, the Human Coronavirus-OC93 was the most common coronavirus with 47.4% of the cases. When comparing demographics and baseline characteristics, both groups were of similar age (sCoV: 74 years vs. COVID-19: 69 years) and presented similar proportions of two or more comorbidities (sCoV: 85.8% vs. COVID-19: 81.6%). More patients with COVID-19 presented with severe disease (78.4% vs. 67.9%), sepsis (36.3% vs. 20.5%), and developed ARDS (15.8% vs. 2.6%) compared to patients with sCoV infection. Patients with COVID-19 had an almost fourfold increased risk of in-hospital death than patients with sCoV infection (OR 3.86, CI 1.99-7.49; p < .001). CONCLUSION: Hospitalized patients with COVID-19 had similar demographics and baseline characteristics to hospitalized patients with sCoV infection; however, patients with COVID-19 presented with higher disease severity, had a higher case-fatality rate, and increased risk of death than patients with sCoV. Clinical findings alone may not help confirm or exclude the diagnosis of COVID-19 during high acute respiratory illness seasons. The respiratory multiplex panel by PCR that includes SARS-CoV-2 in conjunction with local epidemiological data may be a valuable tool to assist clinicians with management decisions.

Efficient delivery of a DNA aptamer-based biosensor into plant cells for glucose sensing through thiol-mediated uptake.

DNA aptamers have been widely used as biosensors for detecting a variety of targets. Despite decades of success, they have not been applied to monitor any targets in plants, even though plants are a major platform for providing oxygen, food, and sustainable products ranging from energy fuels to chemicals, and high-value products such as pharmaceuticals. A major barrier to progress is a lack of efficient methods to deliver DNA into plant cells. We herein report a thiol-mediated uptake method that more efficiently delivers DNA into Arabidopsis and tobacco leaf cells than another state-of-the-art method, DNA nanostructures. Such a method allowed efficient delivery of a glucose DNA aptamer sensor into Arabidopsis for sensing glucose. This demonstration opens a new avenue to apply DNA aptamer sensors for functional studies of various targets, including metabolites, plant hormones, metal ions, and proteins in plants for a better understanding of the biodistribution and regulation of these species and their functions.

A nationwide retrospective analysis of ventilator-associated pneumonia in the US.

Ventilator-associated pneumonia (VAP) is a major cause of healthcare-associated mortality and morbidity in critically ill patients who are mechanically ventilated. The purpose of this study was to describe the various primary discharge diagnoses of hospitalizations with VAP, to identify their demographic characteristics, and to identify risk factors for mortality in hospitalizations with VAP. Hospitalizations with a diagnosis of VAP with mechanical ventilation for over 24 hours were selected from the National Inpatient Sample in 2016 and 2017. In total, 33,140 hospitalizations with VAP were analyzed. The leading principal discharge diagnoses for hospitalizations leading to VAP were sepsis due to an unspecified organism (16.92%), respiratory failure (8.09%), and VAP (6.38%). Mortality among hospitalizations with VAP was 20.9%. Independent risk factors for mortality in hospitalizations with VAP were uninsured status (adjusted odds ratio [aOR] 2.13, 95% confidence interval [CI] 1.49-3.06, P < 0.001), acute renal failure (aOR 2.00, 95% CI 1.75-2.30, P < 0.001), and liver disease (aOR 1.82, 95% CI 1.52-2.18, P < 0.001). In conclusion, VAP is associated with significant mortality. Infective, traumatic, cardiovascular, and respiratory conditions accounted for over 85% of hospitalizations with VAP. Acute renal failure, the presence of liver disease, and lack of insurance are associated with higher mortality in hospitalizations with VAP.

Activity-based photoacoustic probe for biopsy-free assessment of copper in murine models of Wilson's disease and liver metastasis.

The development of high-performance photoacoustic (PA) probes that can monitor disease biomarkers in deep tissue has the potential to replace invasive medical procedures such as a biopsy. However, such probes must be optimized for in vivo performance and exhibit an exceptional safety profile. In this study, we have developed PACu-1, a PA probe designed for biopsy-free assessment (BFA) of hepatic Cu via photoacoustic imaging. PACu-1 features a Cu(I)-responsive trigger appended to an aza-BODIPY dye platform that has been optimized for ratiometric sensing. Owing to its excellent performance, we were able to detect basal levels of Cu in healthy wild-type mice as well as elevated Cu in a Wilson's disease model and in a liver metastasis model. To showcase the potential impact of PACu-1 for BFA, we conducted two blind studies in which we were able to successfully identify Wilson's disease animals from healthy control mice in each instance.

Chemical Composition and Molecular-Specific Optical Properties of Atmospheric Brown Carbon Associated with Biomass Burning.

This study provides molecular insights into the light absorption properties of biomass burning (BB) brown carbon (BrC) through the chemical characterization of tar condensates generated from heated wood pellets at oxidative and pyrolysis conditions. Both liquid tar condensates separated into "darker oily" and "lighter aqueous" immiscible phases. The molecular composition of these samples was investigated using reversed-phase liquid chromatography coupled with a photodiode array detector and a high-resolution mass spectrometer. The results revealed two sets of BrC chromophores: (1) common to all four samples and (2) specific to the "oily" fractions. The common BrC chromophores consist of polar, monoaromatic species. The oil-specific BrC chromophores include less-polar and nonpolar polyaromatic compounds. The most-light-absorbing pyrolysis oily phase (PO) was aerosolized and size-separated using a cascade impactor to compare the composition and optical properties of the bulk versus the aerosolized BrC. The mass absorption coefficient (MAC300-500 nm) of aerosolized PO increased compared to that of the bulk, due to gas-phase partitioning of more volatile and less absorbing chromophores. The optical properties of the aerosolized PO were consistent with previously reported ambient BB BrC measurements. These results suggest the darkening of atmospheric BrC following non-reactive evaporation that transforms the optical properties and composition of aged BrC aerosols.

An overlooked cause of septic shock: Staphylococcal Toxic Shock Syndrome secondary to an axillary abscess.

Staphylococcal Toxic Shock Syndrome (TSS) is characterized by rapid onset of fever, rash, hypotension, and multiorgan system involvement. Clinical manifestations of staphylococcal TSS include fever, chills, hypotension, and a diffuse macular erythroderma followed by desquamation one to two weeks later. The disease came to public attention in the 1980s with the occurrence of a series of menstrual-associated cases. However, the relative incidence of staphylococcal TSS not associated with menstruation has increased, and still, it remains an overlooked cause of septic shock. We present the case of a healthy 19-year-old male that presented with fever, chills, malaise, near-syncope, and a non-fluctuant, mobile nodule in the left armpit. The patient developed septic shock requiring critical care. He underwent extensive investigations resulting negative except for PCR for the detection of MRSA, raising the suspicion for STSS. For that reason, antibiotics for staphylococcal coverage were started, after which he started to improve. Ultimately, the mobile nodule evolved to fluctuant access. Incision and drainage was performed, and cultures confirmed the presence of Staphylococcus aureus.