INTRODUCTION AND AIMS: The risk factors for fatty liver disease are frequent in the Mexican population and the available studies appear to underestimate its prevalence. Our aims were to know the prevalence of metabolic (dysfunction) associated fatty liver disease (MAFLD) in an open population and determine the clinical and sociodemographic characteristics, nutritional status, physical activity level, and prevalence of metabolic syndrome, in the affected population. MATERIALS AND METHODS: Volunteers from the general public were invited to take part in the study. Three separate questionnaires were applied to 1) determine the sociodemographic characteristics and health status of the participants, 2) evaluate the quality of their diet, and 3) to evaluate their level of physical activity. The participants underwent somatometry, laboratory tests, liver ultrasound, and FIB-4 index determination, and transition elastography (FibroScan®) was carried out on all subjects with signs suggestive of liver fibrosis that agreed to undergo the procedure. The statistical analysis was carried out using SPSS for Windows, version 22, software. RESULTS: A total of 585 volunteers were studied, resulting in a prevalence of MAFLD of 41.3%, a predominance of men above 50 years of age, poor dietary habits, and sedentary lifestyle. Male sex, obesity, metabolic syndrome, and elevated ALT were risk factors for the disease and 40% of those affected had advanced fibrosis. CONCLUSIONS: The prevalence of MAFLD in our population is one of the highest in the world. The conditioning factors of the disease can be modified. Therefore, public policies that redirect the current trend are required.
Although cationic polymers are widely used for DNA delivery, the relationship between the properties of the formed complexes and their biological activity is not fully understood. Here, we propose a novel procedure consisting of superresolved images coupled with quantitative colocalization to analyse DNA release in living cells. This work compares the different workflows available in a quantitative colocalization study of DNA delivery using polyethylenimine as transfection reagent. A nimble workflow with deconvolution in three-dimensional images was developed. Among the different colocalization coefficients, Manders' colocalization coefficient was the best to track the complexes. Results showed that DNA/polyethylenimine complexes were tightly interacting at the time of transfection and their disassembly was observed between 2 and 10 h after their uptake. Heterogenicity was found in the intracellular fate of each complex. At 24 h, some complexes were still present underneath the nuclear envelope. Overall, this study opens the door for particle tracking assessment with three-dimensional imaging at intracellular level. LAY DESCRIPTION: DNA delivery technologies in living cells are of high relevance in the biotechnology field. The transient expression of a gene of interest enables the production of a wide range of new therapeutic candidates for clinical purposes. However, the introduction of an exogenous DNA construct into a cell culture requires the use of certain vehicles that protect the DNA from host cell DNases and deliver it into the cell nucleus. From the different systems available, polyethylenimine (PEI) has been extensively used in transient gene expression strategies for the last three decades. However, the intracellular fate of the formed DNA/PEI complexes and the DNA release from the complexes is still poorly understood. In this work, we propose the application of combined superresolved images through mathematical deconvolution to colocalization studies of DNA/PEI complexes evolution in living mammalian cell cultures. Both specimens were covalently labelled with Cy3 and Cy5 dye, respectively, and the kinetics of its disassembly process within the cells was tracked over the time. Because of the specific features of the formed-complexes, a comparative study of the different colocalization coefficients was performed towards optimizing the analysis of these particles with confocal microscopy. Besides, the 3D imaging of the process allowed the direct visualization of a partial DNA/PEI complexes disassembly and the location of those complexes underneath the nuclear envelope during the cell production phase (24 h after the uptake).
Cations/chemistry , DNA/chemistry , Polyethyleneimine/chemistry , Transfection/methods , Cell Nucleus/genetics , Cell Nucleus/ultrastructure , Gene Expression , HEK293 Cells , Humans , Imaging, Three-Dimensional , Microscopy, Confocal , Staining and Labeling
Polyethyleneimine (PEI) has been used extensively for transient gene expression (TGE) in mammalian cell cultures. However, the relationship between DNA/PEI complex preparation and their biological activity has not been fully established. Here, a systematic study of DNA/PEI complexes, their physicochemical properties during formation and their transfection efficiency was performed on a virus-like particle (VLP) production platform. The same chemically defined cell culture medium for DNA/PEI complex formation was used as an alternative to simple ionic solutions to minimize changes in complex properties during transfection. Upon formation, an initial concentration of 1E + 10 DNA/PEI complexes/mL underwent partial aggregation with an average size of 300 nm. The participation of NaCl ions in the evolution of complexes was analyzed by X-ray spectroscopy, stressing the relevance of complexing media composition in TGE strategies. After 15 min incubation, 250 complexes plus aggregates per cell were estimated at the time of transfection. Such heterogeneous preparations cannot be easily characterized; subsequently, nanoparticle tracking analysis (NTA) and cryo-electron microscopy were combined to achieve a complete picture of the preparation. Finally, the contribution of each DNA/PEI complex subpopulation was tested by drug inhibition endocytosis. Interestingly, all complexes delivered DNA efficiently and high size aggregates, which enter through macropinocytosis, when inhibited presented a major contribution to transfection efficiency. There is a need to understand the physicochemical factors that participate in DNA delivery protocols. Hence, this study provides new insights into the characterization of DNA/PEI complexes that will assist in more productive and reproducible TGE strategies.
Chemical Phenomena , DNA/chemistry , Mammals/metabolism , Polyethyleneimine/chemistry , Transfection/methods , Animals , DNA/ultrastructure , Endocytosis , Green Fluorescent Proteins/metabolism , HEK293 Cells , HIV-1/metabolism , Humans , Hydrodynamics , Kinetics , Molecular Weight , gag Gene Products, Human Immunodeficiency Virus/metabolism
Virus-like particles (VLPs) are nanostructures that resemble the structures of viruses. They are composed of one or more structural proteins that can be arranged in several layers and can also contain a lipid outer envelope. VLPs trigger a high humoral and cellular immune response due to their repetitive structures. A key factor regarding VLP safety is the lack of viral genomic material, which enhances safety during both manufacture and administration. Contemporary VLP production may take advantage of several systems, including bacterial, yeast, insect and mammalian cells. The choice of production platform depends on several factors, including cost and the need for post-translational modifications (PTMs), which can be essential in generating an optimal immune response. Some VLP-based vaccines designed to prevent several infectious diseases are already approved and on the market, with many others at the clinical trial or research stage. Interest in this technology has recently increased due to its advantages over classical vaccines. This paper reviews the state-of-the-art of VLP production systems and the newest generation of VLP-based vaccines now available.
Biotechnology/methods , Viral Vaccines/metabolism , Virion/metabolism , Animals , Cell Culture Techniques , Humans , Plants/metabolism
BACKGROUND: Patients with pollinosis develop symptoms after intake of plant food more often than the general population. In order to study the prevalence of the presentation of allergic symptoms to plant foods in pollinosis, we selected a representative sample of the population from our Mediterranean area. METHODS: All patients completed a questionnaire, provided a blood sample and underwent a battery of skin and other complementary tests (prick-prick, oral challenge test) when necessary. The pollen counts were obtained from the Elche pollen station. In addition, sera from a subgroup of patients were checked with an allergen molecule panel on an Advia Centaur XP platform. RESULTS: Of the final sample (n = 233), 39.9% of the patients with pollinosis were sensitized and 30.9% had clinical allergy to at least one of the plant foods studied. Regression analysis showed that age and sensitization to the extracts of Platanus acerifolia and Artemisia vulgaris were the most important variables for discriminating between groups. Patients with pollinosis at a risk of allergy to plant foods had significantly higher Pru p 3 values [odds ratio (OR) 3.3; 95% confidence interval (95% CI) 2.3-4.8], and the value increased according to the number of plant food sensitizations. CONCLUSION: Plant food allergy is more common in patients with pollinosis than in the general population. The use of the London plain tree (P. acerifolia) and mugwort (A. vulgaris) in the skin tests may help identify such patients in our Mediterranean area, but determination of rPru p 3 could also be very useful in patients suspected of having plant food allergy.
Artemisia/immunology , Food Hypersensitivity/immunology , Pollen/immunology , Rhinitis, Allergic, Seasonal/immunology , Trees/immunology , White People , Adolescent , Adult , Aged , Allergens/immunology , Artemisia/chemistry , Comorbidity , Female , Food Hypersensitivity/complications , Food Hypersensitivity/ethnology , Humans , Male , Mediterranean Region/epidemiology , Middle Aged , Prevalence , Regression Analysis , Rhinitis, Allergic, Seasonal/complications , Rhinitis, Allergic, Seasonal/ethnology , Skin Tests , Trees/chemistry
BACKGROUND: Nail disorders are frequent among the geriatric population and unfortunately, repeat avulsion procedures often result in complications. OBJECTIVE: To utilize a phenol-based total matricectomy technique for permanent nail ablation and evaluate its effectiveness. METHODS: We performed total nail ablation on 34 toenails of the hallux (30 patients) with a gauze-phenol application technique and evaluated the degree of pain reported by the patient 12 months postoperatively compared with the preoperative stage. Furthermore, we evaluated the patients' satisfaction with the final cosmetic appearance as well as their satisfaction with the overall procedure. RESULTS: All patients reported 'too much' or 'severe' pain prior to surgery and 94.11% reported 'no pain' 12 months post-surgery. A vast majority of the patients (82.35%) felt pleased with the cosmetic results and all patients reported that they were 'very satisfied' or 'strongly satisfied' with the procedure when interviewed at their 12 month follow-up examination. LIMITATIONS: There were a limited number of participants in this study across an extensive timeframe. CONCLUSION: Phenol-based total nail ablation with specific gauze application is safe, inexpensive and has a high rate of patient satisfaction.
Ablation Techniques/methods , Nail Diseases/surgery , Nails/surgery , Phenols , Follow-Up Studies , Humans , Pain Measurement , Patient Satisfaction , Treatment Outcome
Pretendemos evaluar la utilidad de la detección de antígenode Streptococcus pneumoniae en orina en el diagnóstico de laneumonía extrahospitalaria en niños que necesitaron ingresohospitalarioMATERIAL Y MÉTODOSEstudio prospectivo durante 15 meses de 61 niños ingresadospor neumonía extrahospitalaria y 21 niños sanos como control.La detección del antígeno se realizó mediante el sistemaBinax Now previa concentración de la orinaRESULTADOSLa técnica mostró una sensibilidad del 100%, una especificidaddel 21.4% El 78.8% de los niños con neumonía presentanantígeno en orina, aunque también se encontraba en el 47%de niños sanos (p=0.005). CONCLUSIONESEstá técnica no es útil en el diagnóstico de este proceso por suescasa especificidad. Sin embargo, la dificultad de establecer unpatrón diagnóstico de la neumonía neumocócica no bacteriémicadificulta la correcta evaluación de esta técnica y otros datossugieren que los datos de especificidad pueden ser superiores
Our aim is to assess the usefulness of the detection ofStreptococcus pneumoniae antigen in urine in the diagnosis ofextrahospital pneumonia in children who require admission tohospitalMATERIAL AND METHODSProspective study over a period of 15 months of 61 childrenadmitted to hospital with extrahospital pneumonia and 21healthy children as controls. The antigen was detected usingthe Binax Now system after concentrating the urineRESULTSThe technique was seen to have a sensitivity of 100%, aspecificity of 21.4%. The antigen in urine was present in 78.8%of the children with pneumonia, and also in 47% of the healthychildren (p=0.005). This technique is not useful because it has a low specificity,but the difficulty in establishing diagnostic guidelines for nonbacteraemiapneumococcus pneumonia makes it difficult toassess this technique properly, although other data indicatethat it may have greater specificity
Male , Female , Infant, Newborn , Infant , Child, Preschool , Child , Humans , Streptococcus pneumoniae/isolation & purification , Pneumococcal Infections/urine , Pneumonia/diagnosis , Antigens/isolation & purification , Hospitalization , Community-Acquired Infections/diagnosis , Prospective Studies
Los antifúngicos tópicos se utilizan para el tratamiento de micosis superficiales. En la actualidad se está produciendo un aumento en el consumo de estos fármacos en España, que ha pasado de 8 millones de unidades en 1992 a 10,8 millones en 2000, representando un 65 por ciento del consumo total de antifúngicos. Esto supone pasar de un gasto de 4000 millones de pesetas en 1992 a 6000 millones de pesetas en 2000, lo que representa un 52,4 por ciento del gasto total en antifúngicos. Dentro de los antifúngicos tópicos, el más consumido es ketoconazol, con un 27 por ciento, seguido por clotrimazol y miconazol, con un 17 por ciento y un 14 por ciento, respectivamente. En este mercado, acaparado por los azoles, se puede observar en los últimos años un descenso en el consumo de sertaconazol, flutrimazol y ketoconazol, en beneficio de otro tipo de antifúngicos como ciclopirox, que alcanza en la actualidad un 4,9 por ciento. Otros tipos de antifúngicos tópicos se consumen en menor proporción, como terbinafina (2,1 por ciento), nistatina (1,4 por ciento), amorolfina (1,3 por ciento) y naftifina (1,3 por ciento) (AU)
Middle Aged , Adult , Aged , Male , Female , Humans , Solutions , Self Medication , Spain , Drug Utilization Review , Mycoses , Ointments , Antifungal Agents , Candidiasis, Vulvovaginal , Drug Utilization , Dermatomycoses , Administration, Cutaneous , Administration, Topical , Gels
This study evaluated whether certain antimicrobial agents used in the treatment of community-acquired respiratory infection were effective against the pathogen Neisseria meningitidis, whose natural habitat is the nasopharyngeal mucosa. Antimicrobial agents commonly use in primary healthcare (betalactams, cephalosporins and macrolides), quinolones and rifampicin were studied by determining minimal inhibitory and bactericidal concentrations, time-kill curves and postantibiotic effect. All of them showed bactericidal activity 24 h after incubation. We therefore believe that they are able to empirically eliminate the causative agent of meningococcal meningitis. However, the only antimicrobial agents capable of inducing a significant postantibiotic effect in the tested strain were the quinolones, which slowed down the growth of the microorganism for over 1 h.
Community-Acquired Infections/drug therapy , Drug Resistance , Fluoroquinolones , Meningococcal Infections/prevention & control , Neisseria meningitidis/drug effects , Respiratory Tract Infections/drug therapy , Amoxicillin/pharmacology , Anti-Infective Agents/pharmacology , Ciprofloxacin/pharmacology , Clarithromycin/pharmacology , Humans , Meningococcal Infections/drug therapy , Meningococcal Infections/transmission , Microbial Sensitivity Tests , Naphthyridines/pharmacology , Nasopharynx/microbiology , Neisseria meningitidis/isolation & purification , Rifampin/pharmacology , Time Factors
En el presente estudio valoramos la capacidad que tienen ciertos antimicrobianos utilizados en el tratamiento de la infección respiratoria de la comunidad frente a un patógeno como Neisseria meningitidis, cuyo hábitat natural lo constituye la mucosa nasofaríngea. Mediante determinación de las concentraciones mínimas inhibitorias y bactericidas, las curvas de letalidad y el efecto postantibiótico, se estudiaron los antimicrobianos habitualmente administrados de forma empírica en atención primaria (betalactámicos y macrólidos), quinolonas y rifampicina. Todos mostraron in vitro actividad bactericida a las 24 horas del ensayo, y pensamos que son capaces de eliminar empíricamente de la nasofaringe al agente causal de la meningitis meningocócica. No obstante, los únicos antimicrobianos capaces de inducir efecto postantibiótico significativo en la cepa probada son las quinolonas, que retardan el crecimiento del microorganismo en más de una hora (AU)
Humans , Fluoroquinolones , Drug Resistance , Rifampin , Time Factors , Clarithromycin , Community-Acquired Infections , Meningococcal Infections , Naphthyridines , Nasopharynx , Neisseria meningitidis , Respiratory Tract Infections , Anti-Infective Agents , Ciprofloxacin , Amoxicillin , Microbial Sensitivity Tests
Antifungal Agents/therapeutic use , Administration, Cutaneous , Administration, Topical , Adult , Aged , Antifungal Agents/administration & dosage , Candidiasis, Vulvovaginal/drug therapy , Dermatomycoses/drug therapy , Drug Utilization/statistics & numerical data , Drug Utilization Review/statistics & numerical data , Female , Gels , Humans , Male , Middle Aged , Mycoses/drug therapy , Ointments , Self Medication , Solutions , Spain
We have previously observed that intracerebroventricular infusion of a 5-HT2A receptor antisense oligonucleotide for 8 days results in an increase in cortical 5-HT2A receptor sites and an increase in central 5-HT2A receptor function as measured by quantitation of 5-HT2A receptor-mediated headshake behavior (28). Because lesioning serotonergic neurons or chronic administration of 5-HT2A receptor antagonists does not result in an increase in 5-HT2A receptor density or function in the brain, we have taken advantage of this unique upregulation of 5-HT2A receptors following 5-HT2A receptor antisense oligonucleotide infusion to study the modulation of D1 receptor-mediated behaviors by 5-HT2A receptors. Grooming behavior, elicited by acute injection of SKF 38393, was attenuated after chronic ICV infusion of a 5-HT2A receptor antisense oligonucleotide. There was also a reduction in vacuous chewing behavior induced by SKF 38393, which did not reach statistical significance. Other oral behaviors (i.e., tongue protrusions and gnawing at the cage bottom) were not attenuated. An increase in the density of cortical, as well as striatal 5-HT2A receptor sites was observed following chronic antisense oligonucleotide administration. There was no change in striatal D1 dopamine receptors following 5-HT2A receptor antisense oligonucleotide administration. SKF 38393-induced grooming behavior was also attenuated in naive rats pretreated acutely with the 5-HT2 receptor agonist DOI. These results suggest a role for the 5-HT2A receptor in the modulation of D1 receptor function.
Grooming/physiology , Receptors, Dopamine D1/physiology , Receptors, Serotonin/drug effects , 2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine/pharmacology , Animals , Base Sequence , Corpus Striatum/drug effects , Corpus Striatum/physiology , Dopamine Agonists/pharmacology , Grooming/drug effects , Ketanserin/metabolism , Male , Oligodeoxyribonucleotides, Antisense/pharmacology , Rats , Rats, Sprague-Dawley , Receptor, Serotonin, 5-HT2A , Receptors, Serotonin/genetics , Receptors, Serotonin/physiology , Serotonin Antagonists/metabolism
P11 cells were transfected with DNA for the human 5-hydroxytryptamine1A (5-HT1A) receptor. These cells stably expressed the 5-HT1A receptor coupled to the inhibition of adenylyl cyclase, and not to the stimulation of phosphoinositide hydrolysis. Homologous and heterologous regulation of the 5-HT1A receptor was studied in this cell system. Pretreatment of P11-5HT1A cells with the 5-HT1 receptor agonist 5-carbox-amidotryptamine (5-CT) resulted in a 3-fold increase in both basal and forskolin-stimulated cAMP accumulation, and desensitization of the 5-HT1A receptor as indicated by a decrease in the potency of 8-hydroxydipropylaminotetralin (8-OH-DPAT) to inhibit forskolin-stimulated cAMP accumulation (vehicle-treated cells: EC50 = 2.3 +/- 0.8 nM; 5-CT-treated cells: 9.9 +/- 0.4 nM). The sensitization of adenylyl cyclase as a result of chronic agonist exposure was prevented by the 5-HT1A antagonist WAY100635, which indicated that the effect of 5-CT pretreatment on basal and forskolin-stimulated cAMP accumulation was mediated by 5-HT1A receptor activation. Pretreatment of cells with pertussis toxin abolished the inhibition of forskolin-stimulated cAMP accumulation mediated by 5-HT1A receptor activation and prevented the sensitization of adenylyl cyclase as a result of chronic 5-HT1A receptor agonist exposure. Pretreatment of P11-5HT1A cells with the phorbol ester, phorbol 12-myristate 13-acetate (PMA), also resulted in desensitization of the 5-HT1A receptor, as indicated by a marked decrease in the potency and intrinsic activity of 8-OH-DPAT. No change in the binding characteristics (i.e., Kd or Bmax) of [3H]8-OH-DPAT to 5-HT1A receptor sites was observed after 5-CT or PMA treatments. Activation of alpha-1 adrenergic receptors, but not 5-HT2A receptors, had effects on 5-HT1A receptor responsiveness similar to those seen with PMA pretreatment. In P11-5HT1A cells, homologous regulation of the 5-HT1A receptor was characterized by sensitization of adenylyl cyclase and a decrease in agonist potency, whereas heterologous regulation of the 5-HT1A receptor was characterized by a greater decrease in agonist potency, as well as a marked decrease in intrinsic activity.
Receptors, Serotonin/physiology , 8-Hydroxy-2-(di-n-propylamino)tetralin/metabolism , Adenylyl Cyclases/metabolism , Animals , Cell Line , Humans , Phosphatidylinositols/metabolism , Protein Kinase C/physiology , Rats , Receptors, Serotonin, 5-HT1 , Second Messenger Systems , Serotonin/analogs & derivatives , Serotonin/pharmacology , Serotonin Receptor Agonists/pharmacology , Signal Transduction , Tetradecanoylphorbol Acetate/pharmacology , Transfection
