A fluorogenic substrate for the detection of lipid amidases in intact cells.

Lipid amidases of therapeutic relevance include acid ceramidase (AC), N-acylethanolamine-hydrolyzing acid amidase, and fatty acid amide hydrolase (FAAH). Although fluorogenic substrates have been developed for the three enzymes and high-throughput methods for screening have been reported, a platform for the specific detection of these enzyme activities in intact cells is lacking. In this article, we report on the coumarinic 1-deoxydihydroceramide RBM1-151, a 1-deoxy derivative and vinilog of RBM14-C12, as a novel substrate of amidases. This compound is hydrolyzed by AC (appKm = 7.0 µM; appVmax = 99.3 nM/min), N-acylethanolamine-hydrolyzing acid amidase (appKm = 0.73 µM; appVmax = 0.24 nM/min), and FAAH (appKm = 3.6 µM; appVmax = 7.6 nM/min) but not by other ceramidases. We provide proof of concept that the use of RBM1-151 in combination with reported irreversible inhibitors of AC and FAAH allows the determination in parallel of the three amidase activities in single experiments in intact cells.

Efectividad de la anestesia subaracnoidea con morfina como tratamiento del dolor postoperatorio en cesárea

Fundamento el dolor postoperatorio se considera un dolor con limitaciones de tiempo, a menudo mal controlado. Su manejo representa un gran desafío, ya que la analgesia postoperatoria debe brindar a la madre un control adecuado de este, y a su vez facilitar la atención del bebé. Objetivo evaluar la efectividad de la anestesia subaracnoidea con morfina como tratamiento del dolor postoperatorio en cesárea. Métodos estudio descriptivo y transversal, realizado en el Hospital General Docente Martín Chang Puga, del municipio de Nuevitas, provincia de Camagüey, entre enero de 2021 y diciembre de 2022. La muestra estuvo conformada por 36 pacientes a las cuales se aplicó anestesia subaracnoidea con lidocaína hiperbárica más morfina para la cesárea. Resultados predominó la edad comprendida entre 27-31 años. El 63,9 % de las cesareadas no refirió dolor postoperatorio. Casi la mitad de la población (47,2 %) estudiada presentó efectos secundarios con el uso de la morfina intratecal, principalmente el prurito. El 80,5 % expresó satisfacción con la analgesia postoperatoria. Conclusiones la mayoría de las pacientes encontraron satisfacción con el tratamiento analgésico, a pesar la presencia de efectos adversos, de modo que el uso de morfina intratecal es efectivo en el manejo del dolor poscesárea.

Foundation Postoperative pain is considered time-limited pain, often poorly controlled. Its management represents a great challenge, since postoperative analgesia must provide the mother with adequate control, and at the same time facilitate care for the baby. Objective to evaluate the effectiveness of subarachnoid anesthesia with morphine as a treatment for postoperative pain in cesarean section. Methods descriptive and cross-sectional study carried out at the Martín Chang Puga General Teaching Hospital, in the Nuevitas municipality, Camagüey province, between January 2021 and December 2022. 36 patients to whom subarachnoid anesthesia was applied with Hyperbaric lidocaine plus morphine for cesarean section were considered as the sample. Results the age between 27-31 years predominated. 63.9% of cesarean patients did not report postoperative pain. Almost half of the population (47.2%) studied presented side effects with the use of intrathecal morphine, mainly pruritus. 80.5% expressed satisfaction with postoperative analgesia. Conclusions the majority of patients were satisfied with the analgesic treatment, despite the presence of adverse effects, so that the use of intrathecal morphine is effective in the management of post-cesarean section pain.

IL-1ß-mediated adaptive reprogramming of endogenous human cardiac fibroblasts to cells with immune features during fibrotic remodeling.

The source and roles of fibroblasts and T-cells during maladaptive remodeling and myocardial fibrosis in the setting of pulmonary arterial hypertension (PAH) have been long debated. We demonstrate, using single-cell mass cytometry, a subpopulation of endogenous human cardiac fibroblasts expressing increased levels of CD4, a helper T-cell marker, in addition to myofibroblast markers distributed in human fibrotic RV tissue, interstitial and perivascular lesions in SUGEN/Hypoxia (SuHx) rats, and fibroblasts labeled with pdgfrα CreERt2/+ in R26R-tdTomato mice. Recombinant IL-1ß increases IL-1R, CCR2 receptor expression, modifies the secretome, and differentiates cardiac fibroblasts to form CD68-positive cell clusters. IL-1ß also activates stemness markers, such as NANOG and SOX2, and genes involved in dedifferentiation, lymphoid cell function and metabolic reprogramming. IL-1ß induction of lineage traced primary mouse cardiac fibroblasts causes these cells to lose their fibroblast identity and acquire an immune phenotype. Our results identify IL-1ß induced immune-competency in human cardiac fibroblasts and suggest that fibroblast secretome modulation may constitute a therapeutic approach to PAH and other diseases typified by inflammation and fibrotic remodeling.

Microalga Broths Synthesize Antibacterial and Non-Cytotoxic Silver Nanoparticles Showing Synergy with Antibiotics and Bacterial ROS Induction and Can Be Reused for Successive AgNP Batches.

The era of increasing bacterial antibiotic resistance requires new approaches to fight infections. With this purpose, silver-based nanomaterials are a reality in some fields and promise new developments. We report the green synthesis of silver nanoparticles (AgNPs) using culture broths from a microalga. Broths from two media, with different compositions and pHs and sampled at two growth phases, produced eight AgNP types. Nanoparticles harvested after several synthesis periods showed differences in antibacterial activity and stability. Moreover, an evaluation of the broths for several consecutive syntheses did not find relevant kinetics or activity differences until the third round. Physicochemical characteristics of the AgNPs (core and hydrodynamic sizes, Z-potential, crystallinity, and corona composition) were determined, observing differences depending on the broths used. AgNPs showed good antibacterial activity at concentrations producing no or low cytotoxicity on cultured eukaryotic cells. All the AgNPs had high levels of synergy against Escherichia coli and Staphylococcus aureus with the classic antibiotics streptomycin and kanamycin, but with ampicillin only against S. aureus and tetracycline against E. coli. Differences in the synergy levels were also dependent on the types of AgNPs. We also found that, for some AgNPs, the killing of bacteria started before the massive accumulation of ROS.

Manejo anestésico perioperatorio en histerectomía total abdominal por fibroma uterino en paciente con estenosis aórtica severa. Presentación de un caso

Se presenta el caso de una paciente de 35 años, femenina, con antecedentes de presentar un soplo desde la infancia y que debía ser intervenida quirúrgicamente por fibroma uterino. El anestesiólogo, durante la consulta preoperatoria, constató el soplo ya mencionado por lo que se decidió posponer la cirugía e interconsultar con un cardiólogo. El examen ecocardiográfico demostró que la paciente era portadora de una estenosis aórtica severa con fracción de eyección conservada, prueba ergométrica: clase funcional I, sin isquemia. Mediante anestesia general orotraqueal, se realizó histerectomía total abdominal sin complicaciones. Es de vital importancia para el anestesiólogo el manejo perioperatorio de la estenosis aórtica.

The case of a 35-years-old female patient with a history of presenting a murmur since childhood and who had to undergo surgery for uterine fibroid is presented. The anesthesiologist, during the preoperative consultation, verified the aforementioned murmur, so it was decided to postpone surgery and consult with a cardiologist. The echocardiographic examination showed that the patient had severe aortic stenosis with preserved ejection fraction, stress test: functional class I, without ischemia. Using orotracheal general anesthesia, a total abdominal hysterectomy was performed without complications. The perioperative management of aortic stenosis is of vital importance for the anesthesiologist.

Assessing the relationship between energy-related methane emissions and the burden of cardiovascular diseases: a cross-sectional study of 73 countries.

The energy industry significantly contributes to anthropogenic methane emissions, which add to global warming and have been linked to an increased risk of cardiovascular diseases (CVD). This study aims to evaluate the relationship between energy-related methane emissions and the burden of CVD, measured in disability-adjusted life years (DALYs), in 2019. We conducted a cross-sectional analysis of datasets from 73 countries across all continents. The analyzed datasets included information from 2019 on environmental energy-related methane emissions, burden of DALYs due to CVD. The age-standardized prevalence of obesity in adults and life expectancy at birth were retrieved. The relationship between the variables of interest was evaluated using multiple linear regression models. In the multiple model, we observed a positive linear association between methane emissions and the log-transformed count of DALYs related to CVD. Specifically, for each unit increase in energy-related methane emissions, the burden of CVD increased by 0.06% (95% CI 0.03-0.09%, p < 0.001). The study suggests that reducing methane emissions from the energy industry could improve public health for those at risk of CVD. Policymakers can use these findings to develop strategies to reduce methane emissions and protect public health.

Two-Dimensional Self-Assembly Driven by Intermolecular Hydrogen Bonding in Benzodi-7-azaindole Molecules on Au(111).

The control of molecular structures at the nanoscale plays a critical role in the development of materials and applications. The adsorption of a polyheteroaromatic molecule with hydrogen bond donor and acceptor sites integrated in the conjugated structure itself, namely, benzodi-7-azaindole (BDAI), has been studied on Au(111). Intermolecular hydrogen bonding determines the formation of highly organized linear structures where surface chirality, resulting from the 2D confinement of the centrosymmetric molecules, is observed. Moreover, the structural features of the BDAI molecule lead to the formation of two differentiated arrangements with extended brick-wall and herringbone packing. A comprehensive experimental study that combines scanning tunneling microscopy, high-resolution X-ray photoelectron spectroscopy, near-edge X-ray absorption fine structure spectroscopy, and density functional theory theoretical calculations has been performed to fully characterize the 2D hydrogen-bonded domains and the on-surface thermal stability of the physisorbed material.

Insight into the Antifungal Effects of Propolis and Carnosic Acid-Extension to the Pathogenic Yeast Candida glabrata: New Propolis Fractionation and Potential Synergistic Applications.

Fungi have traditionally been considered opportunistic pathogens in primary infections caused by virulent bacteria, protozoan, or viruses. Consequently, antimycotic chemotherapy is clearly less developed in comparison to its bacterial counterpart. Currently, the three main families of antifungals (polyenes, echinocandins, and azoles) are not sufficient to control the enormous increase in life-threatening fungal infections recorded in recent decades. Natural substances harvested from plants have traditionally been utilized as a successful alternative. After a wide screening of natural agents, we have recently obtained promising results with distinct formulations of carnosic acid and propolis on the prevalent fungal pathogens Candida albicans and Cryptococcus neoformans. Here, we extended their use to the treatment against the emerging pathogenic yeast Candida glabrata, which displayed lower susceptibility in comparison to the fungi mentioned above. Taking into account the moderate antifungal activity of both natural agents, the antifungal value of these combinations has been improved through the obtention of the hydroethanolic fractions of propolis. In addition, we have demonstrated the potential clinical application of new therapeutical designs based on sequential pre-treatments with carnosic/propolis mixtures, followed by exposure to amphotericin B. This approach increased the toxic effect induced by this polyene.

Simultaneous Inhibition of Ceramide Hydrolysis and Glycosylation Synergizes to Corrupt Mitochondrial Respiration and Signal Caspase Driven Cell Death in Drug-Resistant Acute Myeloid Leukemia.

Acute myelogenous leukemia (AML), the most prevalent acute and aggressive leukemia diagnosed in adults, often recurs as a difficult-to-treat, chemotherapy-resistant disease. Because chemotherapy resistance is a major obstacle to successful treatment, novel therapeutic intervention is needed. Upregulated ceramide clearance via accelerated hydrolysis and glycosylation has been shown to be an element in chemotherapy-resistant AML, a problem considering the crucial role ceramide plays in eliciting apoptosis. Herein we employed agents that block ceramide clearance to determine if such a "reset" would be of therapeutic benefit. SACLAC was utilized to limit ceramide hydrolysis, and D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-threo-PDMP) was used to block the glycosylation route. The SACLAC D-threo-PDMP inhibitor combination was synergistically cytotoxic in drug-resistant, P-glycoprotein-expressing (P-gp) AML but not in wt, P-gp-poor cells. Interestingly, P-gp antagonists that can limit ceramide glycosylation via depression of glucosylceramide transit also synergized with SACLAC, suggesting a paradoxical role for P-gp in the implementation of cell death. Mechanistically, cell death was accompanied by a complete drop in ceramide glycosylation, concomitant, striking increases in all molecular species of ceramide, diminished sphingosine 1-phosphate levels, resounding declines in mitochondrial respiratory kinetics, altered Akt, pGSK-3ß, and Mcl-1 expression, and caspase activation. Although ceramide was generated in wt cells upon inhibitor exposure, mitochondrial respiration was not corrupted, suggestive of mitochondrial vulnerability in the drug-resistant phenotype, a potential therapeutic avenue. The inhibitor regimen showed efficacy in an in vivo model and in primary AML cells from patients. These results support the implementation of SL enzyme targeting to limit ceramide clearance as a therapeutic strategy in chemotherapy-resistant AML, inclusive of a novel indication for the use of P-gp antagonists.

Single-Electron Charging of Thioctic Acid Monolayer-Protected Gold Clusters.

There is great interest in the use of Monolayer-Protected Gold Clusters (AuMPCs) as nanoscale capacitors in aqueous media for nanobiotechnological applications, such as bioelectrocatalysts, biofuel cells, and biosensors. However, AuMPCs exhibiting subattofarad double-layer capacitance at room temperature, and the resolution of single-electron charging, has been mainly obtained in an organic medium with nonfunctional capping ligands. We report here the synthesis of Thioctic Acid Monolayer-Protected Au Clusters (TA-AuMPCs) showing electrochemical single electron quantized capacitance charging in organic and aqueous solutions and when immobilized onto different self-assembled monolayer-modified gold electrodes. The presence of functional carboxylic groups opens a simple strategy for interfacing a nanoparticle assembly to biomolecules for their use as electron donors or acceptors in biological electron transfer reactions.

Characterization of the external exposome and its contribution to the clinical respiratory and early biological effects in children: The PROMESA cohort study protocol.

BACKGROUND: Air pollution contains a mixture of different pollutants from multiple sources. However, the interaction of these pollutants with other environmental exposures, as well as their harmful effects on children under five in tropical countries, is not well known. OBJECTIVE: This study aims to characterize the external exposome (ambient and indoor exposures) and its contribution to clinical respiratory and early biological effects in children. MATERIALS AND METHODS: A cohort study will be conducted on children under five (n = 500) with a one-year follow-up. Enrolled children will be followed monthly (phone call) and at months 6 and 12 (in person) post-enrolment with upper and lower Acute Respiratory Infections (ARI) examinations, asthma development, asthma control, and genotoxic damage. The asthma diagnosis will be pediatric pulmonologist-based and a standardized protocol will be used. Exposure, effect, and susceptibility biomarkers will be measured on buccal cells samples. For environmental exposures PM2.5 will be sampled, and questionnaires, geographic information, dispersion models and Land Use Regression models for PM2.5 and NO2 will be used. Different statistical methods that include Bayesian and machine learning techniques will be used for the ambient and indoor exposures-and outcomes. This study was approved by the ethics committee at Universidad Pontificia Bolivariana. EXPECTED STUDY OUTCOMES/FINDINGS: To estimate i) The toxic effect of particulate matter transcending the approach based on pollutant concentration levels; ii) The risk of developing an upper and lower ARI, based on different exposure windows; iii) A baseline of early biological damage in children under five, and describe its progression after a one-year follow-up; and iv) How physical and chemical PM2.5 characteristics influence toxicity and children's health.

High-throughput discovery of novel small-molecule inhibitors of acid Ceramidase.

Ceramide has a key role in the regulation of cellular senescence and apoptosis. As Ceramide levels are lowered by the action of acid ceramidase (AC), abnormally expressed in various cancers, the identification of AC inhibitors has attracted increasing interest. However, this finding has been mainly hampered by the lack of formats suitable for the screening of large libraries. We have overcome this drawback by adapting a fluorogenic assay to a 384-well plate format. The performance of this optimised platform has been proven by the screening a library of 4100 compounds. Our results show that the miniaturised platform is well suited for screening purposes and it led to the identification of several hits, that belong to different chemical classes and display potency ranges of 2-25 µM. The inhibitors also show selectivity over neutral ceramidase and retain activity in cells and can therefore serve as a basis for further chemical optimisation.

Tromboembolismo pulmonar de riesgo intermedio en el posoperatorio mediato de apendicectomía: informe de caso

Introducción: El tromboembolismo pulmonar es una enfermedad de relevancia clínica, en pacientes que se le han realizado intervenciones quirúrgicas. La tasa de mortalidad por esta causa ha disminuido en los últimos años. Objetivo: Describir el manejo diagnóstico y terapéutico del tromboembolismo pulmonar de riesgo intermedio. Caso clínico: Paciente masculino de 62 años de edad que se encontraba ingresado por haber sido operado de una apendicitis gangrenosa. En el posoperatorio mediato comenzó con cuadro de dificultad respiratoria, se realizaron exámenes complementarios incluido ecocardiograma transtorácico, donde se evidenció el signo de McConnell, por lo que se diagnosticó un tromboembolismo pulmonar de riesgo intermedio. El paciente se somete al tratamiento trombolítico estandarizado, luego de lo cual la clínica y los parámetros ecocardiográficos que evolucionaron hacia la mejoría. Conclusiones: Se describió el diagnóstico y tratamiento del tromboembolismo pulmonar de riesgo intermedio en el posoperatorio mediato de apendicectomía, donde se evidenció la utilidad del uso del ecocardiograma doppler transtorácico con la identificación de los signos característicos y la aplicación exitosa de trombolisis sistémica.

Introduction: Pulmonary thromboembolism (PTE) is an entity of clinical relevance in patients in settings related to surgical interventions. The mortality rate from this cause has decreased in recent years. Objective: To describe the diagnostic and therapeutic management of intermediate risk pulmonary thromboembolism. Clinical case: A 62-year-old male patient who was hospitalized for having been operated on for gangrenous appendicitis. In the immediate postoperative period, he begins with symptoms of respiratory distress, complementary tests are performed, including a transthoracic echocardiogram, where McConnell's sign is evident, for which an intermediate-risk pulmonary thromboembolism is diagnosed. The patient undergoes standardized thrombolytic treatment, after which the clinical and echocardiographic parameters evolve towards improvement. Conclusions: The diagnosis and treatment of intermediate risk pulmonary thromboembolism in the immediate postoperative period of appendectomy was described. The usefulness of the use of transthoracic Doppler echocardiography was evidenced with the identification of the characteristic signs and the successful application of systemic thrombolysis.

Fluorescently Labeled Ceramides and 1-Deoxyceramides: Synthesis, Characterization, and Cellular Distribution Studies.

Ceramides (Cer) are bioactive sphingolipids that have been proposed as potential disease biomarkers since they are involved in several cellular stress responses, including apoptosis and senescence. 1-Deoxyceramides (1-deoxyCer), a particular subtype of noncanonical sphingolipids, have been linked to the pathogenesis of type II diabetes. To investigate the metabolism of these bioactive lipids, as well as to have a better understanding of the signaling processes where they participate, it is essential to expand the toolbox of fluorescent sphingolipid probes exhibiting complementary subcellular localization. Herein, we describe a series of new sphingolipid probes tagged with two different organic fluorophores, a far-red/NIR-emitting coumarin derivative (COUPY) and a green-emitting BODIPY. The assembly of the probes involved a combination of olefin cross metathesis and click chemistry reactions as key steps, and these fluorescent ceramide analogues exhibited excellent emission quantum yields, being the Stokes' shifts of the COUPY derivatives much higher than those of the BODIPY counterparts. Confocal microscopy studies in HeLa cells confirmed an excellent cellular permeability for these sphingolipid probes and revealed that most of the vesicles stained by COUPY probes were either lysosomes or endosomes, whereas BODIPY probes accumulated either in Golgi apparatus or in nonlysosomal intracellular vesicles. The fact that the two sets of fluorescent Cer probes have such different staining patterns indicates that their subcellular distribution is not entirely defined by the sphingolipid moiety but rather influenced by the fluorophore.

Green Extracellular Synthesis of Silver Nanoparticles by Pseudomonas alloputida, Their Growth and Biofilm-Formation Inhibitory Activities and Synergic Behavior with Three Classical Antibiotics.

Bacterial resistance to antibiotics is on the rise and hinders the fight against bacterial infections, which are expected to cause millions of deaths by 2050. New antibiotics are difficult to find, so alternatives are needed. One could be metal-based drugs, such as silver nanoparticles (AgNPs). In general, chemical methods for AgNPs' production are potentially toxic, and the physical ones expensive, while green approaches are not. In this paper, we present the green synthesis of AgNPs using two Pseudomonas alloputida B003 UAM culture broths, sampled from their exponential and stationary growth phases. AgNPs were physicochemically characterized by transmission electron microscopy (TEM), total reflection X-ray fluorescence (TXRF), infrared spectroscopy (FTIR), dynamic light scattering (DLS), and X-ray diffraction (XRD), showing differential characteristics depending on the synthesis method used. Antibacterial activity was tested in three assays, and we compared the growth and biofilm-formation inhibition of six test bacteria: Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis. We also monitored nanoparticles' synergic behavior through the growth inhibition of E. coli and S. aureus by three classical antibiotics: ampicillin, nalidixic acid, and streptomycin. The results indicate that very good AgNP activity was obtained with particularly low MICs for the three tested strains of P. aeruginosa. A good synergistic effect on streptomycin activity was observed for all the nanoparticles. For ampicillin, a synergic effect was detected only against S. aureus. ROS production was found to be related to the AgNPs' antibacterial activity.

Factors associated with virologic failure in HIV patients on antiretroviral therapy.

OBJECTIVE: To determine the factors associated with virologic failure n HIV  patients on antiretroviral treatment treated in a Colombian health institution. METHOD: This was a cross-sectional observational retrospective analytical study of HIV patients receiving antiretroviral treatment between 2007­2020. Sociodemographic, pharmacological and clinical variables were collected, including viral load, adherence, and the medication  possession ratio. For statistical analysis, crude and adjusted odds ratios and confidence intervals were obtained. RESULTS: In a population of 5,406 patients, the proportion of virologic failure  was 16.7%. Moreover, in the adjusted model, an association was found  between virologic failure and time on treatment greater than one year,  medication possession ratio under 80%, failure to claim medications from the  pharmacy due to dose omission or discontinuation, adherence under 85%, CD4  count under 500, total cholesterol levels above 201 mg/dL, high density  lipoproteins under 39 mg/dL and presence of mycosis. CONCLUSIONS: In our cohort of HIV patients, short treatment periods, CD4 counts under 200, a low medication possession ratio, failure to timely claim medications from the pharmacy due to omission or  discontinuation, and a lower degree of adherence were factors related to  virologic failure.

OBJETIVO: Determinar los factores asociados al fracaso virológico en pacientes  con el virus de la inmunodeficiencia humana con tratamiento antirretroviral  atendidos en una institución de salud colombiana.Método: Estudio transversal, observacional, retrospectivo y analítico en pacientes con el virus de la inmunodeficiencia humana con tratamiento antirretroviral entre 2007-2020. Se recogieron variables  Sociodemográficas, farmacológicas y clínicas, incluyendo la carga viral, el  grado de adherencia y la tasa de reclamación de medicamentos. Para el  análisis estadístico se obtuvieron las odds ratio crudas y ajustadas y los  intervalos de confianza. RESULTADOS: De una población de 5.406 pacientes, la proporción de fracaso  virológico fue de 16,7% y en el modelo ajustado se encontró asociación entre  el fracaso virológico con el tiempo en el tratamiento mayor a un año, la tasa de posesión de la reclamación inferior al 80%, la no reclamación oportuna por  omisión o suspensión, la adherencia inferior al 85%, el recuento de CD4  inferior a 500, niveles de colesterol total mayores de 201 mg/dl, lipoproteína  de alta densidad menor de 39 mg/dl y presencia de micosis. CONCLUSIONES: En nuestra cohorte de pacientes con el virus de la  Inmunodeficiencia humana, un menor tiempo en tratamiento, un recuento de CD4 menor de 200, una baja tasa de reclamación de los medicamentos, así  como la no reclamación oportuna por omisión y suspensión y un menor grado  de adherencia son factores que se relacionan con el fracaso virológico.

Factores asociados al fracaso virológico en pacientes VIH con tratamiento antirretroviral / Factors associated with virologic failure in HIV patients on antiretroviral therapy

Objetivo: Determinar los factores asociados al fracaso virológico enpacientes con el virus de la inmunodeficiencia humana con tratamientoantirretroviral atendidos en una institución de salud colombiana.Método: Estudio transversal, observacional, retrospectivo y analítico enpacientes con el virus de la inmunodeficiencia humana con tratamientoantirretroviral entre 2007-2020. Se recogieron variables sociodemográficas,farmacológicas y clínicas, incluyendo la carga viral, el grado de adherenciay la tasa de reclamación de medicamentos. Para el análisis estadístico seobtuvieron las odds ratio crudas y ajustadas y los intervalos de confianza.Resultados: De una población de 5.406 pacientes, la proporción defracaso virológico fue de 16,7% y en el modelo ajustado se encontró asociación entre el fracaso virológico con el tiempo en el tratamiento mayora un año, la tasa de posesión de la reclamación inferior al 80%, la noreclamación oportuna por omisión o suspensión, la adherencia inferior al85%, el recuento de CD4 inferior a 500, niveles de colesterol total mayores de 201 mg/dl, lipoproteína de alta densidad menor de 39 mg/dl ypresencia de micosis.Conclusiones: En nuestra cohorte de pacientes con el virus de la inmunodeficiencia humana, un menor tiempo en tratamiento, un recuento deCD4 menor de 200, una baja tasa de reclamación de los medicamentos,así como la no reclamación oportuna por omisión y suspensión y un menorgrado de adherencia son factores que se relacionan con el fracaso virológico. (AU)

Objective: To determine the factors associated with virologic failurein HIV patients on antiretroviral treatment treated in a Colombian healthinstitution.Method: This was a cross-sectional observational retrospective analytical study of HIV patients receiving antiretroviral treatment between2007-2020. Sociodemographic, pharmacological and clinical variableswere collected, including viral load, adherence, and the medication possession ratio. For statistical analysis, crude and adjusted odds ratios andconfidence intervals were obtained.Results: In a population of 5,406 patients, the proportion of virologicfailure was 16.7%. Moreover, in the adjusted model, an association wasfound between virologic failure and time on treatment greater than oneyear, medication possession ratio under 80%, failure to claim medications from the pharmacy due to dose omission or discontinuation, adherence under 85%, CD4 count under 500, total cholesterol levels above201 mg/dL, high density lipoproteins under 39 mg/dL and presence ofmycosis.Conclusions: In our cohort of HIV patients, short treatment periods, CD4counts under 200, a low medication possession ratio, failure to timelyclaim medications from the pharmacy due to omission or discontinuation,and a lower degree of adherence were factors related to virologic failure. (AU)

Cell death induction in Aspergillus fumigatus: accentuating drug toxicity through inhibition of the unfolded protein response (UPR).

One of the most potent opportunistic fungal pathogens of humans is Aspergillus fumigatus, an environmental mold that causes a life-threatening pneumonia with a high rate of morbidity and mortality. Despite advances in therapy, issues of drug toxicity and antifungal resistance remain an obstacle to effective therapy. This underscores the need for more information on fungal pathways that could be pharmacologically manipulated to either reduce the viability of the fungus during infection, or to unleash the fungicidal potential of current antifungal drugs. In this review, we summarize the emerging evidence that the ability of A. fumigatus to sustain viability during stress relies heavily on an adaptive signaling pathway known as the unfolded protein response (UPR), thereby exposing a vulnerability in this fungus that has strong potential for future therapeutic intervention.

Methuosis Contributes to Jaspine-B-Induced Cell Death.

Methuosis is a type of programmed cell death in which the cytoplasm is occupied by fluid-filled vacuoles that originate from macropinosomes (cytoplasmic vacuolation). A few molecules have been reported to behave as methuosis inducers in cancer cell lines. Jaspine B (JB) is a natural anhydrous sphingolipid (SL) derivative reported to induce cytoplasmic vacuolation and cytotoxicity in several cancer cell lines. Here, we have investigated the mechanism and signalling pathways involved in the cytotoxicity induced by the natural sphingolipid Jaspine B (JB) in lung adenocarcinoma A549 cells, which harbor the G12S K-Ras mutant. The effect of JB on inducing cytoplasmic vacuolation and modifying cell viability was determined in A549 cells, as well as in mouse embryonic fibroblasts (MEF) lacking either the autophagy-related gene ATG5 or BAX/BAK genes. Apoptosis was analyzed by flow cytometry after annexin V/propidium iodide staining, in the presence and absence of z-VAD. Autophagy was monitored by LC3-II/GFP-LC3-II analysis, and autophagic flux experiments using protease inhibitors. Phase contrast, confocal, and transmission electron microscopy were used to monitor cytoplasmic vacuolation and the uptake of Lucifer yellow to assess macropinocyosis. We present evidence that cytoplasmic vacuolation and methuosis are involved in Jaspine B cytotoxicity over A549 cells and that activation of 5' AMP-activated protein kinase (AMPK) could be involved in Jaspine-B-induced vacuolation, independently of the phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin complex 1 (PI3K/Akt/mTORC1) axis.

Chirality-Puckering correlation and intermolecular interactions in Sphingosines: Rotational spectroscopy of jaspine B3 and its monohydrate.

Chirality is determinant for sphingosine biofunctions and pharmacological activity, yet the reasons for the biological chiral selection are not well understood. Here, we characterized the intra- and intermolecular interactions at the headgroup of the cytotoxic anhydrophytosphingosine jaspine B, revealing chirality-dependent correlations between the puckering of the ring core and the formation of amino-alcohol hydrogen bond networks, both in the monomer and the monohydrate. Following the specific synthesis of a shortened 3-carbon side-chain molecule, denoted jaspine B3, six different isomers were observed in a jet expansion using broadband (chirped-pulsed) rotational spectroscopy. Additionally, a single isomer of the jaspine B3 monohydrate was observed, revealing the insertion of water in between the hydroxy and amino groups and the formation of a network of O-H···N-H···Oring hydrogen bonds. The specific jaspine B3 stereochemistry thus creates a double-faced molecule where the exposed lone-pair electrons may easily catalyze the formation of intermolecular aggregates and determine the sphingosine biological properties.