Methodological Approach to Use Fresh and Cryopreserved Vessels as Tools to Analyze Pharmacological Modulation of the Angiogenic Growth.

The sprouting of new vessels is greatly influenced by the procedure chosen. We sought to optimize the experimental conditions of the angiogenic growth of fresh and cryopreserved vessels cultured in Matrigel with the aim to use this system to analyze the pharmacological modulation of the process. Segments of second-order branches of rat mesenteric resistance arteries, thoracic aorta of rat or mouse, and cryopreserved rat aorta and human femoral arteries were cultured in Matrigel for 7-21 days in different mediums, as well as in the absence of endothelial or adventitia layer. Quantification of the angiogenic growth was performed by either direct measurement of the mean length of the neovessels or by calcein AM staining and determination of fluorescence intensity and area. Fresh and cryopreserved arterial rings incubated in Matrigel exhibited a spontaneous angiogenic response that was strongly accelerated by fetal calf serum. Addition of vascular endothelial growth factor, fibroblast growth factor, endothelial growth factor, or recombinant insulin-like growth factor failed to increase aortic sprouting, unless all were added together. Removal of adventitia, but not the endothelial layer, abrogated the angiogenic response of aortic rings. Determination of the mean neovessel length is an easy and accurate method to quantify the angiogenic growth devoid of confounding factors, such as inclusion of other cellular types surrounding the neovessels. Activity of a α1-adrenoceptor agonist (phenylephrine) and its inhibition by a selective antagonist (prazosin) were analyzed to prove the usefulness of the Matrigel system to evaluate the pharmacological modulation of the angiogenic growth.

Adenosine A2B receptor agonist improves epidermal barrier integrity in a murine model of epidermal hyperplasia.

Adenosine regulates multiple physiological processes through the activation of four receptor subtypes, of which the A2B adenosine receptor (A2BAR) has the lowest affinity for adenosine. Being the adenosine receptor subtype most prominently expressed in epidermis, we recently described the antiproliferative and anti-inflammatory effect of the selective A2BAR agonist BAY60-6583 (BAY) in human keratinocytes stimulated with 12-O-tetradecanoylphorbol-13-acetate (TPA), so we sought to establish the effect of topical application of BAY in a model of murine epidermal hyperplasia. Topical application of BAY (1 or 10 µg/site) prevented the inflammatory reaction and skin lesions induced by TPA, minimizing hyperproliferation and acanthosis, as well as the expression of specific markers of proliferative keratinocytes. On the other hand, pre-treatment with the selective A2BAR antagonist, PSB-1115 (PSB, 5 or 50 µg/site) reversed these beneficial effects. Additionally, BAY application normalized the expression of epidermal barrier proteins, whose integrity is altered in inflammatory skin diseases, while treatment with the antagonist alone worsened it. Our results, besides confirming the anti-inflammatory and antiproliferative effects of the A2BAR agonist, further demonstrate a role of A2BAR activation to preserve the epidermal barrier. Therefore, the activation of A2BAR may constitute a possible new pharmacological target for the treatment of skin inflammatory diseases such as psoriasis.

Picomolar gradients of progesterone select functional human sperm even in subfertile samples.

More than 1 million infertility treatments are practiced around the world per year, but only 30% of the couples succeed in taking a baby home. Reproductive technology depends in part on sperm quality, which influences not only fertilization but also embryo development and implantation. In order to provide a better quality sperm subpopulation, innovative sperm selection techniques based on physiological sperm features are needed. Spermatozoa at an optimum state may be selected by following an increasing concentration gradient of picomolar progesterone, a steroid secreted by the cumulus cells at the time of ovulation. In this study we developed a method to recruit spermatozoa at the best functional state, based on sperm guidance toward progesterone. The sperm selection assay (SSA) consists of a device with two wells connected by a tube. One well was filled with the sperm suspension and the other with picomolar progesterone, which diffused inside the connecting tube as a gradient. The sperm quality after the SSA was analyzed in normal and subfertile semen samples. Several sperm parameters indicative of sperm physiological state were determined before and after the SSA: capacitation, DNA integrity and oxidative stress. After the SSA, the mean level of capacitated spermatozoa increased three times in normal and in subfertile samples. The level of sperm with intact DNA was significantly increased, while sperm oxidative stress was decreased after sperm selection. Interestingly, the exposure to a progesterone gradient stimulated the completion of capacitation in some spermatozoa that could not do it by themselves. Thus, the SSA supplies a sperm population enriched with spermatozoa at an optimum physiological state that may improve the assisted reproductive technology outcome.

Potential antipsoriatic effect of chondroitin sulfate through inhibition of NF-κB and STAT3 in human keratinocytes.

Chondroitin sulfate (CS) is a natural glycosaminoglycan, formed by the 1-3 linkage of d-glucuronic acid to N-acetylgalactosamine, present in the extracellular matrix. It is used as a slow acting disease modifying agent in the treatment of osteoarthritis, and part of its beneficial effects are due to its antiinflammatory properties that result from an inhibitory effect on NF-κB signaling pathway. This ability raises the hypothesis that CS might be effective in other chronic inflammatory processes such as psoriasis, in which a deregulation of NF-κB is a key feature. In addition, psoriasis is characterized by an upregulation of STAT3 signaling pathway that is related to the epidermal hyperplasia. In the present study we report the pharmacological modulation of the NF-κB and STAT3 signaling pathways by CS in normal human keratinocytes. CS inhibited NF-κB activation and the release of some of the key psoriatic cytokines such as TNFα, IL-8, IL-6 and CCL27. Moreover, it impaired STAT3 translocation to the nucleus and significantly reduced STAT3 transcriptional activity by a mechanism that was independent from STAT3 phosphorylation. Our results confirm the interest of CS as a candidate for future drug research in the therapeutics of psoriasis given the need of more effective and safer oral medications for these patients.

Sustained low disease activity measured by ASDAS slow radiographic spinal progression in axial spondyloarthritis patients treated with TNF-inhibitors: data from REGISPONSERBIO.

BACKGROUND: To evaluate the influence of the disease activity on radiographic progression in axial spondyloarthritis (axSpA) patients treated with TNF inhibitors (TNFi). METHODS: The study included 101 axSpA patients from the Spanish Register of Biological Therapy in Spondyloarthritides (REGISPONSERBIO), which had clinical data and radiographic assessment available. Patients were classified into 2 groups based on the duration of TNFi treatment at baseline: (i) long-term treatment (≥4 years) and (ii) no long-term treatment (< 4 years). Radiographs were scored by two readers according to the modified Stoke Ankylosing Spondylitis Spine Score (mSASSS) with known chronology. Disease activity differences between patients' groups at each time point were assessed using a linear mixed-effect model. RESULTS: Radiographic progression was defined as an increase in ≥2 mSASSS units. At inclusion, approximately half of the patients (45.5%) were receiving long-term treatment with TNFi (≥4 years). In this group of subjects, a significant difference in averaged Ankylosing Spondylitis disease Activity Score (ASDAS) across follow-up was found between progressors and non-progressors (2.33 vs 1.76, p=0.027, respectively). In patients not under long-term TNFi treatment (54.5%) though, no significant ASDAS differences were observed between progressors and non-progressors until the third year of follow-up. Furthermore, no significant differences were found in progression status, when disease activity was measured by Bath Ankylosing spondylitis Disease Activity Index (BASDAI) and C reactive protein (CRP). CONCLUSIONS: Patients on long-term TNFi treatment with a mean sustained low disease activity measures by ASDAS presented lower radiographic progression than those with active disease.

Cyanocobalamin Ultraflexible Lipid Vesicles: Characterization and In Vitro Evaluation of Drug-Skin Depth Profiles.

Atopic dermatitis (AD) and psoriasis are the most common chronic inflammatory skin disorders, which importantly affect the quality of life of patients who suffer them. Among other causes, nitric oxide has been reported as part of the triggering factors in the pathogenesis of both conditions. Cyanocobalamin (vitamin B12) has shown efficacy as a nitric oxide scavenger and some clinical trials have given positive outcomes in its use for treating skin pathologies. Passive skin diffusion is possible only for drugs with low molecular weights and intermediate lipophilicity. Unfortunately, the molecular weight and hydrophilicity of vitamin B12 do not predict its effective diffusion through the skin. The aim of this work was to design new lipid vesicles to encapsulate the vitamin B12 to enhance its skin penetration. Nine prototypes of vesicles were generated and characterized in terms of size, polydispersity, surface charge, drug encapsulation, flexibility, and stability with positive results. Additionally, their ability to release the drug content in a controlled manner was demonstrated. Finally, we found that these lipid vesicle formulations facilitated the penetration of cyanocobalamin to the deeper layers of the skin. The present work shows a promising system to effectively administer vitamin B12 topically, which could be of interest in the treatment of skin diseases such as AD and psoriasis.

Annexin A2-Mediated Plasminogen Activation in Endothelial Cells Contributes to the Proangiogenic Effect of Adenosine A2A Receptors.

Adenosine A2A receptor mediates the promotion of wound healing and revascularization of injured tissue, in healthy and animals with impaired wound healing, through a mechanism depending upon tissue plasminogen activator (tPA), a component of the fibrinolytic system. In order to evaluate the contribution of plasmin generation in the proangiogenic effect of adenosine A2A receptor activation, we determined the expression and secretion of t-PA, urokinase plasminogen activator (uPA), plasminogen activator inhibitor-1 (PAI-1) and annexin A2 by human dermal microvascular endothelial cells stimulated by the selective agonist CGS-21680. The plasmin generation was assayed through an enzymatic assay and the proangiogenic effect was studied using an endothelial tube formation assay in Matrigel. Adenosine A2A receptor activation in endothelial cells diminished the release of PAI-1 and promoted the production of annexin A2, which acts as a cell membrane co-receptor for plasminogen and its activator tPA. Annexin A2 mediated the increased cell membrane-associated plasmin generation in adenosine A2A receptor agonist treated human dermal microvascular endothelial cells and is required for tube formation in an in vitro model of angiogenesis. These results suggest a novel mechanism by which adenosine A2A receptor activation promotes angiogenesis: increased endothelial expression of annexin A2, which, in turn, promotes fibrinolysis by binding tPA and plasminogen to the cell surface.

Wound healing is accelerated by agonists of adenosine A2 (G alpha s-linked) receptors.

The complete healing of wounds is the final step in a highly regulated response to injury. Although many of the molecular mediators and cellular events of healing are known, their manipulation for the enhancement and acceleration of wound closure has not proven practical as yet. We and others have established that adenosine is a potent regulator of the inflammatory response, which is a component of wound healing. We now report that ligation of the G alpha s-linked adenosine receptors on the cells of an artificial wound dramatically alters the kinetics of wound closure. Excisional wound closure in normal, healthy mice was significantly accelerated by topical application of the specific A2A receptor agonist CGS-21680 (50% closure by day 2 in A2 receptor antagonists. In rats rendered diabetic (streptozotocin-induced diabetes mellitus) wound healing was impaired as compared to nondiabetic rats; CGS-21680 significantly increased the rate of wound healing in both nondiabetic and diabetic rats. Indeed, the rate of wound healing in the CGS-21680-treated diabetic rats was greater than or equal to that observed in untreated normal rats. These results appear to constitute the first evidence that a small molecule, such as an adenosine receptor agonist, accelerates wound healing in both normal animals and in animals with impaired wound healing.

Improved effectiveness from individualized dosing of self-administered biologics for the treatment of moderate-to-severe psoriasis: a 5-year retrospective chart review from a Spanish University Hospital.

Background: Biologics for moderate-to-severe psoriasis are expensive and treatment substitutions may vastly increase cost. Moreover, administration regimens in routine practice may differ from recommended guidelines.Objectives: To evaluate long-term effectiveness, regimen, drug-survival, and efficiency of self-administered biologics in clinical practice.Methods: We performed a 5-year retrospective study in 72 patients (44 ± 14 years old) with moderate-to-severe psoriasis at the University Hospital La Plana (Vila-real, Spain), treated with subcutaneous biologics. We determined the effectiveness (PASI 75 or PASI < 5), and drug-survival using Kaplan-Meier estimates, and analyzed reasons for treatment interruption, drug substitution patterns, and costs.Results: Etanercept was less effective (45%) than ustekinumab (85%) and adalimumab (71%). In 15% of patients, optimal responses were maintained despite dose intervals lengthening. Drug-survival was significantly lower for etanercept than for the other biologics (p < .005). Most adalimumab and etanercept discontinuations were due to adverse events or lack of effectiveness; for ustekinumab the causes were unrelated to drug effects. Ustekinumab was 100% effective as a secondary biologic.Conclusion: Ustekinumab was the safest and most efficient treatment. Etanercept showed the highest treatment failure rate, incurring higher costs. Dosage individualization according to patient needs improves the therapy efficiency, reducing therapeutic failure and derived costs.

Microneedle-Based Delivery: An Overview of Current Applications and Trends.

Microneedle arrays (MNA) are considered as one of the most promising resources to achieve systemic effects by transdermal delivery of drugs. They are designed as a minimally invasive, painless system which can bypass the stratum corneum, overcoming the potential drawbacks of subcutaneous injections and other transdermal delivery systems such as chemical enhancers, nano and microparticles, or physical treatments. As a trendy field in pharmaceutical and biomedical research, its applications are constantly evolving, even though they are based on very well-established techniques. The number of molecules administered by MNA are also increasing, with insulin and vaccines administration being the most investigated. Furthermore, MNA are being used to deliver cells and applied in other organs and tissues like the eyes and buccal mucosae. This review intends to offer a general overview of the current state of MNA research, focusing on the strategies, applications, and types of molecules delivered recently by these systems. In addition, some information about the materials and manufacturing processes is presented and safety data is discussed.

Trends in Scientific Production on Pharmaceutical Follow-up andthe Dader Method / Tendencias en la producción científica sobre el seguimiento farmacéutico y el método Dader

Objective: Pharmacotherapeutic Follow-up is the Professional Pharmaceutical Care Service aimed at detecting Drug-Related Problems for the prevention and resolution of negative medicine outcomes. The Dader Method is considered a clear and simple tool to develop Pharmacotherapeutic Follow-up. This research aims to analyze the evolution of the international scientific production related to Pharmacotherapeutic Follow-up and the Dader Method to show the current situation of this Professional Pharmacy Assistance Service. In addition, from the data obtained, we give a critical perspective on the implementation of the Dader Method in Community Pharmacy, considering its advantages and disadvantages based on the published scientific literature. Methods: Using bibliometrics tools, indicators were obtained to analyze the international production of scientific articles on Pharmacotherapeutic Followup and the Dader Method during the period (1999-2022) through the Scopus database. Results: The results showed a growth in the international scientific production of publications on Pharmacotherapeutic Follow-up, obtaining 30,287 papers, placing the United States, the United Kingdom, Australia, Canada and Spain as the five most productive countries. The publication of 83 papers on the Dader Method places Spain with the highest number of publications, followed by other Spanish or Portuguese speaking countries, among which Brazil and Colombia have the most prominent number of published papers in Latin America. The most frequent international journal covering the topic of Pharmacotherapeutic Follow-up was the American Journal of Health- Pharmacy (12.4%), while on the Dader Method, the journal Pharmaceutical Care Spain (21.7%) is in the first position, followed by Farmacia Hospitalaria (8.4%). Conclusion: The publications on the Dader method highlighs the greater productivity of the University of Granada and the author María José Faus Dáder. The inclusion of patients in the PTF service using the Dader Method, is more frequent in the hospital context, and is based on the presence of defined chronic pathologies (mainly diabetes), polymedication or specialized care follow-up, with elderly population being the most represented in all cases.(AU)

UPSTAGING, CENTRALITY AND SURVIVAL IN EARLY STAGE NON-SMALL CELL LUNG CANCER VIDEO-ASSISTED SURGERY.

OBJECTIVES: Hiliar (pN1) and mediastinal lymph (pN2) nodal upstaging after surgery for early stage (

Algoritmo para la selección de colgajos perforantes perimamarios en reconstrucción inmediata poscirugía conservadora / Algorithm to select chest wall perforator flaps in immediate conservative breast surgery reconstruction

RESUMEN Antecedentes: los colgajos perforantes perimamarios son de gran utilidad en la reconstrucción mamaria inmediata en cirugía conservadora. Objetivo: describir los resultados del empleo de un algoritmo sobre colgajos perforantes perimamarios en la reconstrucción mamaria inmediata después de cirugía conservadora por cáncer de mama. Material y métodos: se llevó a cabo un estudio retrospectivo descriptivo. Se revisaron las historias clínicas de las pacientes operadas entre enero de 2020 y diciembre de 2022 por carcinoma de mama con cirugía conservadora y que requirieron reconstrucción con colgajos perimamarios. Las indicaciones incluyeron déficit de volumen, defecto de contorno y asimetría. Se evaluó el pedículo vascular del colgajo mediante Doppler color en todos los casos, lo que permitió seguir un algoritmo para la selección de la mejor opción de colgajo. Resultados: se realizaron 20 colgajos en 19 pacientes. Promedio de edad: 52 años ± 11 (rango 30-76). No existieron complicaciones intraoperatorias. Una paciente requirió reoperación por compresión del pedículo vascular del colgajo por hematoma, con la pérdida parcial, y otro colgajo sufrió epidermólisis superficial. No hubo pérdidas totales de ningún colgajo. Todas recibieron radioterapia posoperatoria y no experimentaron pérdida de volumen ni retracciones. Con un promedio de seguimiento de 15 meses, las pacientes valoraron los resultados a 6 meses como excelente en 7, bueno en 11 y regular en 2. Conclusión: la selección de colgajos perforantes locales para corregir defectos mamarios después de cirugía conservadora, mediante el examen con Doppler color preoperatorio para la identificación del pedículo vascular y un algoritmo específico, permitió obtener resultados estéticos satisfactorios sin requerir elementos aloplásticos ni revisiones posteriores.

ABSTRACT Background: Chest wall perforator flaps are a good option for immediate breast reconstruction after conservative surgery. Objective: The aim of this study was to describe the clinical results of an algorithm for using chest wall perforator flaps for breast reconstruction after breast-conserving surgery for breast cancer. Material and methods: We conducted a descriptive and retrospective study. The information was retrieved from the medical records of the patients diagnosed with breast cancer who underwent breast-conserving surgery and required reconstruction using chest wall perforator flaps between January 2020 and March 2022. The indications included volume deficit, contour defect and asymmetry. The vascular pedicle of the flap was evaluated by color Doppler ultrasound in all cases, which allowed us to follow an algorithm for selecting the best flap option. Results: Twenty flaps were made in 19 patients. Mean age: 52 years ± 11 (range 30-76). There were no intraoperative complications. One patient required reoperation due to a hematoma with compression of the vascular pedicle of the flap with partial flap loss, and another flap presented superficial epidermolysis. There were no cases of complete flap loss. All the patients underwent postoperative radiation therapy without loss of volume or retractions. Mean follow-up was 15 months. At 6 months, patients rated the results as excellent, good, and fair in 7, 11, and 2 cases, respectively. Conclusion: The selection of local perforator flaps to correct breast defects after conservative surgery, using preoperative color Doppler ultrasound to identify the vascular pedicle and a specific algorithm, allowed us to obtain satisfactory aesthetic results without the need for alloplastic elements or subsequent revisions.

[Buccal mucosa urethroplasty in anterior urethral strictures]. / Uretroplastia con mucosa oral en estenosis de uretra anterior.

UNLABELLED: Buccal mucosal graft can be used for succesfull repair in both pendulous and bulbar strictures. MATERIAL AND METHODS: We present our experience with buccal mucosal graft repair in 8 patients with onlay patch that varies from 4 to 16 cm. in length. Three pendulous, two bulbar and three panurethral strictures were repaired. These patients were observed for 36 to 60 months. RESULTS: No stricture recurrences were observed. Only one patient had lower lip paresthesia for six months.

Coupling biological detection to liquid chromatography: a new tool in drug discovery.

Procedures to characterize drugs that can be obtained from plant extracts or combinatorial chemistry are tedious, and they consume considerable resources (e.g., animals) and time. Thus, we have looked for a way to streamline this process. We describe here a novel system for the pre-characterization of drugs based on liquid chromatography coupled to biological detection using perifused or perfused organs. This novel system allows the on-line detection of pharmacologically active substances in hydrosoluble mixtures from vegetal extracts or combinatorial chemistry libraries. Depending on the volume of drug solution and concentration of the samples, the procedure can work through either medium pressure liquid chromatography or HPLC, and it enables the fingerprints of drugs to be assessed based on their contractile activity on combinations of different isolated tissues. As an example, we show how the system can identify active fractions from an extract of Stevia rebaudiana Bertoni, an activity that was later associated with rebaudioside N. Coupling liquid chromatography to biological detection offers a rapid way to focus attention on active products in complex samples, mostly from hydrosoluble species, helping to considerably reduce the time and cost of the pre-characterization of drugs.

Methotrexate and sulfasalazine promote adenosine release by a mechanism that requires ecto-5'-nucleotidase-mediated conversion of adenine nucleotides.

We and others have shown that an increased extracellular concentration of adenosine mediates the antiinflammatory effects of methotrexate and sulfasalazine both in vitro and in vivo, but the mechanism by which these drugs increase extracellular adenosine remains unclear. The results of the experiments reported here provide three distinct lines of evidence that adenosine results from the ecto-5'-nucleotidase- mediated conversion of adenine nucleotides to adenosine. First, pretreatment of a human microvascular endothelial cell line (HMEC-1) with methotrexate increases extracellular adenosine after exposure of the pretreated cells to activated neutrophils; the ecto-5'-nucleotidase inhibitor alpha, beta-methylene adenosine-5'-diphosphate (APCP) abrogates completely the increase in extracellular adenosine. Second, there is no methotrexate-mediated increase in extracellular adenosine concentration in the supernate of cells deficient in ecto-5'-nucleotidase, but there is a marked increase in extracellular adenosine concentration in the supernates of these cells after transfection and surface expression of the enzyme. Finally, as we have shown previously, adenosine mediates the antiinflammatory effects of methotrexate and sulfasalazine in the murine air pouch model of inflammation, and injection of APCP, the ecto-5'-nucleotidase inhibitor, abrogates completely the increase in adenosine and the decrement in inflammation in this in vivo model. These results not only show that ecto-5'-nucleotidase activity is a critical mediator of methotrexate- and sulfasalazine-induced antiinflammatory activity in vitro and in vivo but also indicate that adenine nucleotides, released from cells, are the source of extracellular adenosine.

An induction gene trap for identifying a homeoprotein-regulated locus.

An important issue in developmental biology is the identification of homeoprotein target genes. We have developed a strategy based on the internalization and nuclear addressing of exogenous homeodomains, using an engrailed homeodomain (EnHD) to screen an embryonic stem (ES) cell gene trap library. Eight integrated gene trap loci responded to EnHD. One is within the bullous pemphigoid antigen 1 (BPAG1) locus, in a region that interrupts two neural isoforms. By combining in vivo electroporation with organotypic cultures, we show that an already identified BPAG1 enhancer/promoter is differentially regulated by homeoproteins Hoxc-8 and Engrailed in the embryonic spinal cord and mesencephalon. This strategy can therefore be used for identifying and mutating homeoprotein targets. Because homeodomain third helices can internalize proteins, peptides, phosphopeptides, and antisense oligonucleotides, this strategy should be applicable to other intracellular targets for characterizing genetic networks involved in a large number of physiopathological states.

[Psychopathology associated with idiopathic nephrotic syndrome in the pediatric age group]. / Psicopatología del síndrome nefrótico idiopático en la edad pediátrica.

AIM: To analyze the influence of several variables in the genesis of psychological morbidity in children with idiopathic nephrotic syndrome (INS) as compared with healthy controls. METHODS: The Strengths and Difficulties Questionnaire (SDQ) was administered to children with INS (n = 23) and a control group of healthy children (n = 38). The SDQ is a well-known screening instrument for psychological/psychiatric morbidity in childhood. The questionnaire consists of five scales that evaluate emotional symptoms, behavioral problems, hyperactivity, peer problems and prosocial behavior. The total score is calculated by adding all the subscales except the prosocial behavior scale. In children with INS, type of treatment, sex, steroid dose, disease stage, disease duration, and prognosis (number of relapses/disease stage) were also noted. RESULTS: Statistically significant differences were found in two of the scales (emotional symptoms and peer problems), as well as in the total score in children with INS as compared with the control group (p < 0.05). A positive correlation was found between disease duration and total score in the INS group. CONCLUSIONS: Emotional and behavioral problems were more frequent in children with INS. The only variable related to the severity of the symptoms studied in these children was disease duration.

Desarrollo y protocolización de un modelo de optimización de respuestas a las consultas en los centros de información del medicamento / Development and protocolisation of a model for optimising responses to queries at the medicines information centres

INTRODUCCIÓN: los Centros de Información de Medicamento (CIM) tienen como objetivo servir a los farmacéuticos como fuente de información técnica, científica y actualizada de los medicamentos y productos sanitarios para promover el uso racional de estos y mejorar el cuidado y la salud del paciente. El farmacéutico ha de disponer de la mejor información sobre el medicamento para dar un buen servicio profesional.OBJETIVO: elaboración, implantación y validación de un protocolo de respuestas a las consultas recibidas en el CIM que permita el registro normalizado de las preguntas recibidas, así como un modelo optimizado de respuestas basado en la recuperación eficiente de información en las bases de datos especializadas.MATERIALES Y MÉTODOS: estudio observacional retrospectivo de las consultas recibidas y desarrollo de una base de datos propia para el registro de las consultas generadas (CAC). Las consultas se clasificaron siguiendo criterios de categorización a través de 25 ejes (comercialización, indicación, seguridad, posología, etc.), fármaco consultado y grupo ATC al que pertenece. Una vez procesadas las consultas de 7 meses, se procedió a la implantación del protocolo normalizado de resolución de consultas para evaluar su eficiencia.RESULTADOS Y DISCUSIÓN: durante el periodo de tiempo comprendido entre junio de 2021 y enero de 2022 se han registrado 248 consultas, que se han categorizado en base al tipo de consulta y fármaco consultado. En febrero, tras iniciar la validación, se recibieron 38 consultas de las cuales se repitieron 9 fármacos consultados (3,21 %), 10 grupos ATC (3,57 %) y se repitieron las mismas consultas 4 veces (1,43 %). Estos resultados permitieron responder de forma rápida y efectiva a las consultas que se repitieron durante el primer mes de validación. (AU)

Thyroid hormone receptor beta1 gene expression is increased by Dexamethasone at transcriptional level in rat liver.

Triiodothyronine (T3) exerts most of its effect through nuclear thyroid hormone receptors (TR) which bind mainly as heterodimers with retinoid-X receptors (RXR) to thyroid hormone response elements (TRE) in target genes. It is well known that the synergistic interaction of T3 and glucocorticoids has a role on the synthesis of growth hormone in rat pituitary cell lines and in the T3-induced metamorphosis in amphibians. Glucocorticoids increased mRNAs of T3-regulated hepatic genes. Our laboratory reported increased specific metabolic actions of T3 in rat liver by Dexamethasone (Dex) through a mechanism involving an up-regulation of the maximal binding capacity of TR. In this study we further explored the participation of TR in the molecular mechanism of the Dex-induced increase on liver T3-specific metabolic action. Dex administration to adrenalectomized rats induced an increase of liver TRbeta1 protein and mRNA. Nuclear run-on assay revealed that Dex up-regulated the TR gene transcriptional rate. Transfection assay in COS-7 cells indicated that Dex increased the transcriptional activity of the TRbeta1 promoter. Electrophoretic mobility shift assay demonstrated that Dex induced the binding of additional proteins related to or neighboring the DNA sequence of a glucocorticoid receptor (GR) binding (GRE) half-site in the TRbeta1 promoter. Evidences for an interaction of GR on the TRbeta1 promoter have been obtained. Moreover, the specificity of the GR binding to GRE was determined not only by the GRE DNA sequence, but also by the interaction of the GR with other transacting factors bound to sequences flanking the GRE.