RNA-sequencing of Human Kidney Allografts and Delineation of T-Cell Genes, Gene Sets, and Pathways Associated With Acute T Cell-mediated Rejection.

BACKGROUND: Delineation of T-cell genes, gene sets, pathways, and T-cell subtypes associated with acute T cell-mediated rejection (TCMR) may improve its management. METHODS: We performed bulk RNA-sequencing of 34 kidney allograft biopsies (16 Banff TCMR and 18 no rejection [NR] biopsies) from 34 adult recipients of human kidneys. Computational analysis was performed to determine the differential intragraft expression of T-cell genes at the level of single-gene, gene set, and pathways. RESULTS: T-cell signaling pathway gene sets for plenary T-cell activation were overrepresented in TCMR biopsies compared with NR biopsies. Heightened expression of T-cell signaling genes was validated using external TCMR biopsies. Pro- and anti-inflammatory immune gene sets were enriched, and metabolism gene sets were depleted in TCMR biopsies compared with NR biopsies. Gene signatures of regulatory T cells, Th1 cells, Th2 cells, Th17 cells, T follicular helper cells, CD4 tissue-resident memory T cells, and CD8 tissue-resident memory T cells were enriched in TCMR biopsies compared with NR biopsies. T-cell exhaustion and anergy were also molecular attributes of TCMR. Gene sets associated with antigen processing and presentation, and leukocyte transendothelial migration were overexpressed in TCMR biopsies compared with NR biopsies. Cellular deconvolution of graft infiltrating cells by gene expression patterns identified CD8 T cell to be the most abundant T-cell subtype infiltrating the allograft during TCMR. CONCLUSIONS: Our delineation of intragraft T-cell gene expression patterns, in addition to yielding new biological insights, may help prioritize T-cell genes and T-cell subtypes for therapeutic targeting.

Transcriptomic signatures of chronic active antibody-mediated rejection deciphered by RNA sequencing of human kidney allografts.

Natural killer (NK) cells mediate spontaneous cell-mediated cytotoxicity and antibody-dependent cell-mediated cytotoxicity. This dual functionality could enable their participation in chronic active antibody-mediated rejection (CA-ABMR). Earlier microarray profiling studies have not subcategorized antibody-mediated rejection into CA-ABMR and active-ABMR, and the gene expression pattern of CA-ABMR has not been compared with that of T cell-mediated rejection (TCMR). To fill these gaps, we RNA sequenced human kidney allograft biopsies categorized as CA-ABMR, active-ABMR, TCMR, or No Rejection (NR). Among the 15,910 genes identified in the biopsies, 60, 114, and 231 genes were uniquely overexpressed in CA-ABMR, TCMR, and active-ABMR, respectively; compared to NR, 50 genes were shared between CA-ABMR and active-ABMR, and 164 genes between CA-ABMR and TCMR. The overexpressed genes were annotated to NK cells and T cells in CA-ABMR and TCMR, and to neutrophils and monocytes in active-ABMR. The NK cell cytotoxicity and allograft rejection pathways were enriched in CA-ABMR. Genes encoding perforin, granzymes, and death receptor were overexpressed in CA-ABMR versus active-ABMR but not compared to TCMR. NK cell cytotoxicity pathway gene set variation analysis score was higher in CA-ABMR compared to active-ABMR but not in TCMR. Principal component analysis of the deconvolved immune cellular transcriptomes separated CA-ABMR and TCMR from active-ABMR and NR. Immunohistochemistry of kidney allograft biopsies validated a higher proportion of CD56+ NK cells in CA-ABMR than in active-ABMR. Thus, CA-ABMR was exemplified by the overexpression of the NK cell cytotoxicity pathway gene set and, surprisingly, molecularly more like TCMR than active-ABMR.

Effects of atipamezole on selected physiologic parameters in cynomolgus macaques (Macaca fascicularis).

BACKGROUND: Atipamezole, an α-2 adrenergic receptor antagonist, reverses the α-2 agonist anesthetic effects. There is a dearth of information on the physiological effects of these drugs in cynomolgus macaques (Macaca fascicularis). We assessed atipamezole's physiologic effects. We hypothesized atipamezole administration would alter anesthetic parameters. METHODS: Five cynomolgus macaques were sedated with ketamine/dexmedetomidine intramuscularly, followed 45 min later with atipamezole (0.5 mg/kg). Anesthetic parameters (heart rate, blood pressure [systolic (SAP), diastolic (DAP), and mean (MAP) blood pressure], body temperature, respiratory rate, and %SpO2) were monitored prior to and every 10 min (through 60 min) post atipamezole injection. RESULTS: While heart rate was significantly increased for 60 min; SAP, DAP, MAP, and temperature were significantly decreased at 10 min. CONCLUSIONS: This study indicates subcutaneous atipamezole results in increased heart rate and transient blood pressure decrease. These findings are clinically important to ensure anesthetist awareness to properly support and treat patients as needed.

Light wavelength and its impact on broiler health.

Light is a powerful management tool in poultry production systems, affecting productivity, physiology, and behavior. The objective of this study was to understand the impacts of three light colors (blue, green, or white) on broiler health. Broilers (N = 14,256) were raised in floor pens with fresh litter from 0 to 35 d in 9 rooms (2 blocked trials). Additionally, 2 genotypes (Ross YPMx708 and EPMx708) and sex were studied (6 room replications per lighting treatment and 18 pen replicates per sex × genotype × lighting program). Blood samples and tissue samples from the retina and the pineal gland were collected from birds (16-18 d of age) 9 times in one 24-hr period per trial, then analyzed to determine melatonin levels (pg/mL). Mobility was assessed via gait scoring, using a 0 to 5 scale at 31 to 32 d of age. Footpad dermatitis was assessed using a 0 to 4 scale, and litter quality by a subjective scoring system (scores ranging from 0-4). Mortality and morbidity causes were identified through necropsies performed by pathologists. Data were analyzed as a 3 × 2 × 2 factorial design, with trial as a random variable block and lighting treatment nested within rooms (MIXED procedure, SAS). Birds raised under blue light had lower serum melatonin levels during one time-point during the scotophase, but no other differences were noted. No effect of light color was observed for melatonin produced in the tissues, nor mobility and footpad dermatitis. An interaction was noted for litter quality where a higher percentage of pens housing YPM-708 broilers had litter categorized into dry, but not easily moved with the foot (category 1). Males had higher incidence of infectious and metabolic deaths than females. Interactions were observed between light and sex, where males raised under white light had a higher incidence of skeletal causes of mortality. Overall, the results showed that light color had minor impacts only on melatonin levels, mobility, footpad dermatitis, litter quality, and cause of mortality.

Light color and the commercial broiler: effect on behavior, fear, and stress.

Light is an important component in poultry production, and it may impact bird behavior, an important component of animal welfare. Light-emitting diode (LED) lamps are of interest for broiler production since they are inexpensive to run and provide monochromatic colors. This study aimed to understand the impact of three light colors (blue, green, or white), provided by LED lighting, on behavioral expression, stress and fear levels of broilers. A total of 14,256 male and female broilers of 2 genotypes (Ross EPMx708 and Ross YPMx708) were housed in 9 rooms in 2 blocked trials (3 room replicates per light per trial), with sexes and genotypes housed in 12 separate pens per room. Behavioral expression was recorded using an infrared camera and analyzed using a scan sampling technique. To assess fear, 3 tests were conducted: tonic immobility, novel object, and response to observer. Blood was collected to evaluate chronic stress using the heterophil:lymphocyte (H:L) ratio. Data were statistically analyzed using SAS (MIXED procedure) in a 3 × 2 × 2 factorial design, with lighting treatment nested within room. Fear tests indicated reduced fear levels in birds raised under blue light (lower latency to rise during the tonic immobility test and a lower percentage of birds moving due to the passage by of an observer). No differences were observed for the novel object test. Light color resulted in changes in stress levels, indicated by a lower H:L ratio for broilers raised under blue light compared to those raised under white light. Behavior was influenced by light color, especially at 33 to 34 d of age, where birds raised under white light were more active, and birds raised under blue light spent more time resting. Overall, results indicated that light color has minor influences on behavioral expression. Utilizing blue light during the brooding and rearing phase leads to lower stress and a reduction in fear, suggesting that blue light may improve the emotional states of fear and stress, thereby improving the welfare of poultry.

Does light color during brooding and rearing impact broiler productivity?

Light color during brooding and rearing may impact broiler production; however, literature results are inconsistent. To address this, the effects of 3 wavelength spectra on broiler performance in 2 sex and 2 genotypes (Ross YPMx708 and EPMx708) were studied. Broilers were raised (d 0-35) under wavelength programs provided by LED light bulbs (blue (455 nm), green (510 nm) or white) under similar intensities (clux). Two trials were conducted (total number of birds =  14,256; 6 room replications per lighting treatment; 18 replicate pens per light × sex × genotype). Data were analyzed as a 3 × 2 × 2 (wavelength × sex × genotype) factorial design, with trial as a random variable block and wavelength nested within rooms (Proc Mixed, SAS 9.4). Birds raised under white light were heavier than under blue or green light at d7 (P = 0.004), and green at d14 (P = 0.03). Feed intake, gain-to-feed efficiency and flock uniformity (d15, 28) did not differ. Mortality only differed at wk 5, when broilers raised under white light had higher mortality than those raised under blue (P = 0.03). YPM-708 were heavier at 21 d (P = 0.007), 28 d (P = 0.001), and 35 d (P < 0.0001), had a better total feed conversion rate (P < 0.0001), higher mortality for wk 1 (P = 0.001), lower mortality during the last wk (P = 0.02) and better uniformity at 28 d (P = 0.01) than EPM-708 broilers. Males were heavier at all measured ages except d0 (d7-P = 0.03, other weeks P < 0.0001), had better total feed conversion (P < 0.0001), increased weekly mortality except for wk 1 (wk2-P = 0.04, wk3-P = 0.002, wk4, 5, and total-P = 0.0001) and were less uniform (P = 0.0002) than females. YPM-708 and EPM-708 males had higher total feed intake (P < 0.0001), and males raised under white light had higher mortality than females raised under white or blue light (P = 0.01). To conclude, the use of specific light colors (blue and green) had only minor impacts on broiler production when light intensity was equalized and balanced for bird spectral sensitivity, and its use to improve productivity does not appear to be advantageous for broilers in a commercial setting.

Biologically synthesized silver nanoparticles as potent antibacterial effective against multidrug-resistant Pseudomonas aeruginosa.

Pseudomonas aeruginosa is one of the most worrisome infectious bacteria due to its intrinsic and acquired resistance against several antibiotics and the recalcitrance of its infections; hence, the development of novel antimicrobials effective against multidrug-resistant P. aeruginosa is mandatory. In this work, silver nanoparticles obtained by green synthesis using a leaf extract and fungi were tested against a battery of clinical strains from cystic fibrosis, pneumonia and burnt patients, some of them with multidrug resistance. Both nanoparticles showed a potent antibacterial effect, causing severe damage to the cell wall, membrane and DNA, and inducing the production of reactive oxygen species. Moreover, the nanoparticles derived from fungi showed synergistic antibacterial effects with the antibiotics meropenem and levofloxacin for some clinical strains and both kinds of nanoparticles were nontoxic for larvae of the moth Galleria mellonella, encouraging further research for their implementation in the treatment of P. aeruginosa infections.

Detection of infiltrating fibroblasts by single-cell transcriptomics in human kidney allografts.

We tested the hypothesis that single-cell RNA-sequencing (scRNA-seq) analysis of human kidney allograft biopsies will reveal distinct cell types and states and yield insights to decipher the complex heterogeneity of alloimmune injury. We selected 3 biopsies of kidney cortex from 3 individuals for scRNA-seq and processed them fresh using an identical protocol on the 10x Chromium platform; (i) HK: native kidney biopsy from a living donor, (ii) AK1: allograft kidney with transplant glomerulopathy, tubulointerstitial fibrosis, and worsening graft function, and (iii) AK2: allograft kidney after successful treatment of active antibody-mediated rejection. We did not study T-cell-mediated rejections. We generated 7217 high-quality single cell transcriptomes. Taking advantage of the recipient-donor sex mismatches revealed by X and Y chromosome autosomal gene expression, we determined that in AK1 with fibrosis, 42 months after transplantation, more than half of the kidney allograft fibroblasts were recipient-derived and therefore likely migratory and graft infiltrative, whereas in AK2 without fibrosis, 84 months after transplantation, most fibroblasts were donor-organ-derived. Furthermore, AK1 was enriched for tubular progenitor cells overexpressing profibrotic extracellular matrix genes. AK2, eight months after successful treatment of rejection, contained plasmablast cells with high expression of immunoglobulins, endothelial cell elaboration of T cell chemoattractant cytokines, and persistent presence of cytotoxic T cells. In addition to these key findings, our analysis revealed unique cell types and states in the kidney. Altogether, single-cell transcriptomics yielded novel mechanistic insights, which could pave the way for individualizing the care of transplant recipients.

Decreased levels of cathepsin Z mRNA expressed by immune blood cells: diagnostic and prognostic implications in prostate cancer.

Cathepsin Z (CTSZ) is a cysteine protease responsible for the adhesion and migration of both immune and tumor cells. Due to its dual role, we hypothesized that the site of CTSZ expression could be determinant of the pro- or anti-tumorigenic effects of this enzyme. To test this hypothesis, we analyzed CTSZ expression data in healthy and tumor tissues by bioinformatics and evaluated the expression levels of CTSZ mRNA in the blood cells of prostate cancer (PCa) patients by qRT-PCR compared with healthy subjects, evaluating its diagnostic and prognostic implications for this type of cancer. Immune cells present in the blood of healthy patients overexpress CTSZ. In PCa, we found decreased CTSZ mRNA levels in blood cells, 75% lower than in healthy subjects, that diminished even more during biochemical relapse. CTSZ mRNA in the blood cells had an area under the curve for PCa diagnosis of 0.832, with a 93.3% specificity, and a positive likelihood ratio of 9.4. The site of CTSZ mRNA expression is fundamental to determine its final role as a protective determinant in PCa, such as CTSZ mRNA in the blood cells, or a malignant determinant, such as found for CTSZ expressed in high levels by different types of primary and metastatic tumors. Low CTSZ mRNA expression in the total blood is a possible PCa marker complementary to prostate-specific antigen (PSA) for biopsy decisions, with the potential to eliminate unnecessary biopsies.

High-quality intake of carbohydrates is associated with lower prevalence of subclinical atherosclerosis in femoral arteries: The AWHS study.

BACKGROUND AND AIMS: High-quality of the carbohydrates consumed, apart from their total amount, appear to protect from cardiovascular disease (CVD). However, the relationship between the quality of carbohydrates and the early appearance of atherosclerosis has not yet been described. Our objective was to estimate the association between the quality of dietary carbohydrates and subclinical atherosclerosis in femoral and carotid arteries. METHODS: Cross-sectional study of femoral and carotid atherosclerosis assessed using ultrasounds of 2074 middle-aged males, 50.9 (SD 3.9) years old, with no previous CVD, and pertaining to the Aragon Workers' Health Study (AWHS) cohort. Food frequency questionnaires were used to calculate a carbohydrate quality index (CQI) defined as: consumption of dietary fiber, a lower glycemic index, the ratio of whole grains/total grains, and the ratio of solid carbohydrates/total carbohydrates. The presence of plaques across four CQI intervals was studied using adjusted logistic regression models. RESULTS: The CQI showed a direct inverse association with subclinical atherosclerosis in femoral territories. Participants with a higher consumption of high-quality carbohydrates (13-15 points) were less likely to have femoral plaques when compared with participants in the lowest index interval (4-6 points) (OR = 0.59; 95% CI = 0.39, 0.89; p = 0.005). No association was found between the CQI and the presence of subclinical atherosclerosis in carotid territories. A lower consumption of high-quality carbohydrates tended to be associated with a greater atherosclerosis extension, considered as the odds for having more affected territories (p = 0.011). CONCLUSIONS: Among middle-aged males, a high-quality intake of carbohydrates is associated with a lower prevalence of femoral artery subclinical atherosclerosis when compared with a lower consumption. Thus, indicating an early relationship between the quality of carbohydrates and the development of CVD.

Ubiquitous atmospheric production of organic acids mediated by cloud droplets.

Atmospheric acidity is increasingly determined by carbon dioxide and organic acids1-3. Among the latter, formic acid facilitates the nucleation of cloud droplets4 and contributes to the acidity of clouds and rainwater1,5. At present, chemistry-climate models greatly underestimate the atmospheric burden of formic acid, because key processes related to its sources and sinks remain poorly understood2,6-9. Here we present atmospheric chamber experiments that show that formaldehyde is efficiently converted to gaseous formic acid via a multiphase pathway that involves its hydrated form, methanediol. In warm cloud droplets, methanediol undergoes fast outgassing but slow dehydration. Using a chemistry-climate model, we estimate that the gas-phase oxidation of methanediol produces up to four times more formic acid than all other known chemical sources combined. Our findings reconcile model predictions and measurements of formic acid abundance. The additional formic acid burden increases atmospheric acidity by reducing the pH of clouds and rainwater by up to 0.3. The diol mechanism presented here probably applies to other aldehydes and may help to explain the high atmospheric levels of other organic acids that affect aerosol growth and cloud evolution.

Antibacterial properties of phenothiazine derivatives against multidrug-resistant Acinetobacter baumannii strains.

AIM: As options to treat recalcitrant bacterial infections which are increasingly limited due to multidrug-resistant strains, searching for new, effective antibacterial compounds is necessary. One strategy is to generate treatment alternatives by drug repurposing. METHODS AND RESULTS: In this work, phenotypic microarrays were used for the screening of miscellaneous compounds against the growth and biofilm formation of Acinetobacter baumannii, an important emergent multidrug-resistant opportunistic pathogen. The results showed that the phenothiazine derivatives, such as promethazine, trifluoperazine, thioridazine, and chlorpromazine, inhibited the growth of antibiotic-sensitive and multidrug-resistant strains (showing minimal inhibitory concentrations ranging from 0·05 to 0·6 g l-1 and minimal bactericidal concentrations ranging from 0·1 to 2·5 g l-1 ). All phenothiazine derivatives were active against biofilm cells (with minimal biofilm eradication concentrations ranging from 0·5 to >3 g l-1 ). Chlorpromazine promoted reactive oxigen species (ROS) production, and cell membrane and DNA damage. Chlorpromazine showed synergy with antibiotics such as ceftazidime, meropenem, and colistin and was an effective treatment for experimentally infected Galleria mellonella when combined with ceftazidime. CONCLUSIONS: It was demonstrated that phenothiazine derivatives, especially chlorpromazine, are drugs with attractive antibacterial properties against nosocomial MDR strains of A. baumannii, by generating ROS and cell membrane and DNA damage. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study indicates that repurposing phenothiazine derivatives for treating recalcitrant infections by A. baumannii could be promising.

FOXP3 mRNA Profile Prognostic of Acute T Cell-mediated Rejection and Human Kidney Allograft Survival.

BACKGROUND: T cell-mediated rejection (TCMR) is the most frequent type of acute rejection and is associated with kidney allograft failure. Almost 40% of TCMR episodes are nonresponsive to therapy, and molecular mechanisms for the nonresponsiveness are unknown. Our single-center study identified that urinary cell FOXP3 mRNA abundance predicts TCMR reversibility and allograft survival. METHODS: We developed PCR assays and measured absolute copy numbers of transcripts for FOXP3, CD25, CD3E, perforin, and 18S rRNA in 3559 urines from 480 kidney allograft recipients prospectively enrolled in the multicenter Clinical Trials in Organ Transplantation-04. In this replication study, we investigated the association between mRNA profile and TCMR diagnosis, TCMR reversibility, and allograft survival. RESULTS: 18S rRNA normalized levels of mRNA for FOXP3 (P = 0.01, Kruskal-Wallis test), CD25 (P = 0.01), CD3E (P < 0.0001), and perforin (P < 0.0001) were diagnostic of TCMR, but only FOXP3 mRNA level predicted TCMR reversibility (ROC AUC = 0.764; 95% confidence interval, 0.611-0.917; P = 0.008). Multivariable logistic regression analyses showed that urinary cell FOXP3 mRNA level predicted reversal, independent of clinical variables. A composite model of clinical variables and FOXP3 mRNA (AUC = 0.889; 95% CI, 0.781-0.997; P < 0.001) outperformed FOXP3 mRNA or clinical variables in predicting TCMR reversibility (P = 0.01, likelihood ratio test). Multivariable Cox proportional hazards regression analyses showed that FOXP3 mRNA level predicts kidney allograft survival (P = 0.047) but not after controlling for TCMR reversal (P = 0.477). CONCLUSIONS: Urinary cell level of FOXP3 mRNA is diagnostic of TCMR, predicts TCMR reversibility, and is prognostic of kidney allograft survival via a mechanism involving TCMR reversal.

Impact of a multidisciplinary medication reconciliation program on clinical outcomes: A pre-post intervention study in surgical patients.

BACKGROUND: Previous studies have evaluated the effects of medication reconciliation (MR) and suggest that it is effective in decreasing medication discrepancies. Nevertheless, a recent overview of systematic reviews concluded that there is no clear evidence in favor of MR in patient-related outcomes and healthcare utilization, and further research about it is needed. OBJECTIVE: To evaluate the impact of a multidisciplinary MR program on clinical outcomes in patients with colorectal cancer presenting other chronic diseases, undergoing elective colorectal surgery. METHODS: We performed a pre-post study. Adult patients scheduled for elective colorectal cancer surgery were included if they presented at least one "high-risk" criteria. The MR program was developed by internists, pharmacists and surgeons, and ended with the obtention of the patient's pre-admission medication list and follow-up care until discharge. The primary outcome was the length of stay (LOS). Secondly, we evaluated mortality, preventable surgery cancellations and risk factors for complications. RESULTS: Three hundred and eight patients were enrolled. Only one patient in the pre-intervention group suffered a preventable surgery cancellation (p = 0.317). The mean LOS was 13 ± 12 vs. 11 ± 5 days in the pre-intervention and the intervention cohort, respectively (p = 0.435). A difference in favor of the intervention group in patients with cardiovascular disease (p = 0.038) and those >75 years old (p = 0.043) was observed. No difference was detected in the mortality rate (p = 0.999) neither most of the indicators of risk factors for complications. However, the management of preoperative systolic blood pressure of hypertensive patients (p = 0.004) and insulin reconciliation in patients with treated diabetes (p = 0.003) were statistically better in the intervention group. CONCLUSIONS: No statistically significant change was observed in the mean global LOS. A statistically significant positive effect on LOS was observed in vulnerable populations: patients >75 years old and those with cardiovascular disease, who presented a 5-day reduction in the mean LOS.

Decreased levels of cathepsin Z mRNA expressed by immune blood cells: diagnostic and prognostic implications in prostate cancer

Cathepsin Z (CTSZ) is a cysteine protease responsible for the adhesion and migration of both immune and tumor cells. Due to its dual role, we hypothesized that the site of CTSZ expression could be determinant of the pro- or anti-tumorigenic effects of this enzyme. To test this hypothesis, we analyzed CTSZ expression data in healthy and tumor tissues by bioinformatics and evaluated the expression levels of CTSZ mRNA in the blood cells of prostate cancer (PCa) patients by qRT-PCR compared with healthy subjects, evaluating its diagnostic and prognostic implications for this type of cancer. Immune cells present in the blood of healthy patients overexpress CTSZ. In PCa, we found decreased CTSZ mRNA levels in blood cells, 75% lower than in healthy subjects, that diminished even more during biochemical relapse. CTSZ mRNA in the blood cells had an area under the curve for PCa diagnosis of 0.832, with a 93.3% specificity, and a positive likelihood ratio of 9.4. The site of CTSZ mRNA expression is fundamental to determine its final role as a protective determinant in PCa, such as CTSZ mRNA in the blood cells, or a malignant determinant, such as found for CTSZ expressed in high levels by different types of primary and metastatic tumors. Low CTSZ mRNA expression in the total blood is a possible PCa marker complementary to prostate-specific antigen (PSA) for biopsy decisions, with the potential to eliminate unnecessary biopsies.

COVID-19 with and without pneumonia: clinical outcomes in the internal medicine ward / COVID-19 con y sin neumonía: resultados en la sala de medicina interna

Abstract The clinical features of COVID-19 differ substantially upon the presence (or absence) of viral pneumonia. The aim of this article was to describe the clinical characteristics of COVID-19 patients admitted to the Internal Medicine ward, as divided into those with and without pneumonia. This single-center prospective cohort study was conducted in a tertiary teaching public hospital in Buenos Aires City named Hospital General de Agudos Carlos G. Durand. Baseline data collection was performed within 48 hours of admission and patients were followed until discharge or in-hospital death. Epidemiological, clinical, laboratory, and radiological characteristics together with treatment data were obtained from the medical records. Of the 417 included, 243 (58.3%) had pneumonia. Median age was 43 years (IQR:32-57) and 222 (53.2%) were female. The overall crude case-fatality rate was 3.8%. None of the COVID-19 patients without pneumonia developed critical disease, required invasive mechanical ventilation nor died during hospitalization. However, 7 (4%) developed severe disease during follow-up. Among patients with COVID-19 pneumonia, in-hospital mortality rate was 6.6%, severe disease developed in 81 (33.3%), critical disease in 23 (9.5%), and 22 (9.1%) were admitted to the intensive care unit. A largely good prognosis was observed among COVID-19 patients without pneumonia, still, even among this group, unfavorable clinical progression can develop and should be properly monitored. Critical illness among patients with COVID-19 pneumonia was frequent and observed rates from this cohort provide a sound characterization of COVID-19 clinical features in a major city from South America.

Resumen Las características clínicas del COVID-19 difieren sustancialmente según la presencia (o ausencia) de neumonía viral. El objetivo de este artículo fue describir las características clínicas de los pacientes con COVID-19 internados en el servicio de Clínica Médica, divididos en pacientes con y sin neumonía. Fue un estudio de cohorte prospectivo, con base en un único centro, realizado en un hospital público de la ciudad de Buenos Aires: Hospital General de Agudos Carlos G. Durand. La recolección basal de datos se realizó dentro de las 48 horas del ingreso y los pacientes fueron seguidos hasta el alta o la muerte hospitalaria. Las características epidemiológicas, clínicas, de laboratorio y radiológicas junto con los datos del tratamiento se obtuvieron de la historia clínica. De los 417 incluidos, 243 (58.3%) tenían neumonía. La mediana de edad fue de 43 años (RIC: 32-57) y 222 (53.2%) eran mujeres. La tasa global de letalidad fue del 3.8%. Ninguno de los pacientes con COVID-19 sin neumonía desarrolló enfermedad crítica, requirió ventilación mecánica invasiva ni falleció durante la hospitalización. Sin embargo, 7 (4%) desarrollaron enfermedad grave durante el seguimiento. Entre aquellos con neumonía COVID-19, la tasa de mortalidad hospitalaria fue del 6.6%, se desarrolló enfermedad grave en 81 (33.3%), enfermedad crítica en 23 (9.5%) y 22 (9.1%) fueron trasladados a la unidad de cuidados intensivos. Los pacientes con COVID-19 sin neumonía presentaron buen pronóstico; sin embargo, incluso en este grupo, se observaron algunos con progresión clínica desfavorable, por lo que se requirió seguimiento adecuado. En los pacientes con neumonía por COVID-19, el desarrollo de enfermedad crítica fue frecuente y las tasas observadas en esta cohorte proporcionan una caracterización sólida de las características clínicas de los pacientes con COVID-19 en una importante ciudad de América del Sur.

AI (Artificial Intelligence) and Hypertension Research.

PURPOSE OF REVIEW: This review a highlights that to use artificial intelligence (AI) tools effectively for hypertension research, a new foundation to further understand the biology of hypertension needs to occur by leveraging genome and RNA sequencing technology and derived tools on a broad scale in hypertension. RECENT FINDINGS: For the last few years, progress in research and management of essential hypertension has been stagnating while at the same time, the sequencing of the human genome has been generating many new research tools and opportunities to investigate the biology of hypertension. Cancer research has applied modern tools derived from DNA and RNA sequencing on a large scale, enabling the improved understanding of cancer biology and leading to many clinical applications. Compared with cancer, studies in hypertension, using whole genome, exome, or RNA sequencing tools, total less than 2% of the number cancer studies. While true, sequencing the genome of cancer tissue has provided cancer research an advantage, DNA and RNA sequencing derived tools can also be used in hypertension to generate new understanding how complex protein network, in non-cancer tissue, adapts and learns to be effective when for example, somatic mutations or environmental inputs change the gene expression profiles at different network nodes. The amount of data and differences in clinical condition classification at the individual sample level might be of such magnitude to overwhelm and stretch comprehension. Here is the opportunity to use AI tools for the analysis of data streams derived from DNA and RNA sequencing tools combined with clinical data to generate new hypotheses leading to the discovery of mechanisms and potential target molecules from which drugs or treatments can be developed and tested. Basic and clinical research taking advantage of new gene sequencing-based tools, to uncover mechanisms how complex protein networks regulate blood pressure in health and disease, will be critical to lift hypertension research and management from its stagnation. The use of AI analytic tools will help leverage such insights. However, applying AI tools to vast amounts of data that certainly exist in hypertension, without taking advantage of new gene sequencing-based research tools, will generate questionable results and will miss many new potential molecular targets and possibly treatments. Without such approaches, the vision of precision medicine for hypertension will be hard to accomplish and most likely not occur in the near future.

Estudio de cohorte prospectivo de pacientes con COVID-19 hospitalizados en servicio de clínica médica del Hospital Durand Protocolo de estudio / Prospective cohort study of patients with COVID-19 hospitalized in the Internal Medicine ward of Hospital Durand: study protocol

INTRODUCCIÓN: Conocer los predictores de mala evolución en pacientes con Enfermedad por Coronavirus 2019 (COVID-19) permite identificar de forma temprana a los pacientes con peor pronóstico, aportando mejores herramientas a la hora de tomar decisiones clínicas. Se presenta el protocolo de un estudio de cohorte cuyo objetivo principal es identificar factores de riesgo de infección severa, critica y mortalidad en pacientes con COVID-19 internados en el Servicio de Clínica Médica del Hospital Durand (Buenos Aires, Argentina). MÉTODOS: Estudio de cohorte prospectivo con base en un único centro. Se incluirá a todos los pacientes que ingresen al servicio de Clínica Médica con diagnóstico de COVID-19 durante el periodo de estudio. Se recolectarán las características epidemiológicas, clínicas, de laboratorio, radiológicas y los datos de tratamiento, al ingreso y al momento del alta o muerte hospitalaria. El evento final primario es la muerte en la internación; los eventos secundarios son el desarrollo de enfermedad grave y enfermedad crítica, la internación en unidad cerrada y el requerimiento de asistencia respiratoria mecánica.

Urinary cell transcriptomics and acute rejection in human kidney allografts.

BACKGROUNDRNA sequencing (RNA-Seq) is a molecular tool to analyze global transcriptional changes, deduce pathogenic mechanisms, and discover biomarkers. We performed RNA-Seq to investigate gene expression and biological pathways in urinary cells and kidney allograft biopsies during an acute rejection episode and to determine whether urinary cell gene expression patterns are enriched for biopsy transcriptional profiles.METHODSWe performed RNA-Seq of 57 urine samples collected from 53 kidney allograft recipients (patients) with biopsies classified as acute T cell-mediated rejection (TCMR; n = 22), antibody-mediated rejection (AMR; n = 8), or normal/nonspecific changes (No Rejection; n = 27). We also performed RNA-Seq of 49 kidney allograft biopsies from 49 recipients with biopsies classified as TCMR (n = 12), AMR (n = 17), or No Rejection (n = 20). We analyzed RNA-Seq data for differential gene expression, biological pathways, and gene set enrichment across diagnoses and across biospecimens.RESULTSWe identified unique and shared gene signatures associated with biological pathways during an episode of TCMR or AMR compared with No Rejection. Gene Set Enrichment Analysis demonstrated enrichment for TCMR biopsy signature and AMR biopsy signature in TCMR urine and AMR urine, irrespective of whether the biopsy and urine were from the same or different patients. Cell type enrichment analysis revealed a diverse cellular landscape with an enrichment of immune cell types in urinary cells compared with biopsies.CONCLUSIONSRNA-Seq of urinary cells and biopsies, in addition to identifying enriched gene signatures and pathways associated with TCMR or AMR, revealed genomic changes between TCMR and AMR, as well as between allograft biopsies and urinary cells.