Battery-free, lightweight, injectable microsystem for in vivo wireless pharmacology and optogenetics.

Pharmacology and optogenetics are widely used in neuroscience research to study the central and peripheral nervous systems. While both approaches allow for sophisticated studies of neural circuitry, continued advances are, in part, hampered by technology limitations associated with requirements for physical tethers that connect external equipment to rigid probes inserted into delicate regions of the brain. The results can lead to tissue damage and alterations in behavioral tasks and natural movements, with additional difficulties in use for studies that involve social interactions and/or motions in complex 3-dimensional environments. These disadvantages are particularly pronounced in research that demands combined optogenetic and pharmacological functions in a single experiment. Here, we present a lightweight, wireless, battery-free injectable microsystem that combines soft microfluidic and microscale inorganic light-emitting diode probes for programmable pharmacology and optogenetics, designed to offer the features of drug refillability and adjustable flow rates, together with programmable control over the temporal profiles. The technology has potential for large-scale manufacturing and broad distribution to the neuroscience community, with capabilities in targeting specific neuronal populations in freely moving animals. In addition, the same platform can easily be adapted for a wide range of other types of passive or active electronic functions, including electrical stimulation.

Aplicación de la Terapia Dialéctico-Conductual en el tratamiento del Trastorno Límite de la Personalidad y de la Patología Dual / The Application of Dialectical Behavior Therapy to the treatment of Borderline Personality Disorder and of Dual Disorder / Aplicació de laTeràpiaDialècticaConductual en el tractamentdel Trastorn Límitde laPersonalitati de laPatologiaDual

En este artículo, se introduce el modelo de hospitalización parcial de adolescentes de la Fundació Orienta (seguido en sus hospitales de día), así como las estrategias terapéuticas empleadas en patologías específicas. Se presenta la Terapia Dialéctico Conductual y se valora su idoneidad en el tratamiento de pacientes adolescentes con Trastorno Límite de Personalidad, así como con Patología Dual. Se describe finalmente la aplicación de dicha terapia en los hospitales de día de la Fundación

The Application of Dialectical Behavior Therapy to the treatment of Borderline Personality Disorder and of Dual Disorder. Fundació Orienta’s adolescent partial hospitalization model (employed in the foundation’s day hospitals) is introduced, as well as the program’s therapeutic strategies for specific pathologies. Dialectical Behavioral Therapy is presented, and its suitability in the treatment of adolescent patients with Borderline Personality Disorder and with Dual Disorder is assessed. Finally, the application of the treatment in Fundació Orienta day hospitals is described

En aquest article s’introdueix el model d’hospitalització parcial d’adolescents de la Fundació Orienta (seguit en els seus hospitals de dia), així com les estratègies terapèutiques utilitzades en patologies específiques. Es presenta la Teràpia Dialèctica Conductual i se’n valora la idoneïtat en el tractament de pacients adolescents amb Trastorn Límit de la Personalitat, així com de Patologia Dual. Es descriu finalment l’aplicació d’aquesta teràpia en els hospitals de dia de la Fundació

Manejo farmacológico perioperatorio en pacientes con glaucoma / Perioperative pharmacological management in patients with glaucoma

PROPÓSITO DE LA REVISIÓN: Ante la necesidad de realizar una facoemulsificación, una cirugía filtrante o la combinación de ambas, pueden plantearse dudas sobre la conveniencia de mantener determinados fármacos antiglaucomatosos. El objetivo del presente trabajo es unificar criterios que puedan orientar la práctica clínica diaria y que permitan desarrollar algoritmos de actuación en el preoperatorio y el postoperatorio de la cirugía filtrante o de catarata. Protocolos propuestos. En el preoperatorio de la cirugía de catarata, el uso de antiinflamatorios no esteroideos queda a criterio del cirujano, recomendándose el formato de monodosis. Se plantea la suspensión de las prostaglandinas unos días antes de la cirugía. Los fármacos sin conservantes favorecen la mejor recuperación de la superficie ocular (SO) tras la cirugía de catarata. Una vez eliminados todos los aspectos modificadores de la presión intraocular (PIO), se debe reevaluar la PIO basal, prefiriendo los fármacos hipotensores sin conservantes, en caso de necesitarlos. La utilización de hipotensores oculares y corticoides libres de conservantes en el preoperatorio de la cirugía de glaucoma reduce el riesgo de fracaso quirúrgico. Se recomienda interrumpir las prostaglandinas. En el postoperatorio de la cirugía de glaucoma los corticoides constituyen el tratamiento antiinflamatorio de elección, siendo preferibles aquellos libres de conservantes. Al reintroducir un tratamiento antiglaucomatoso, se deben evitar los conservantes para no potenciar la cicatrización. CONCLUSIONES: el presente protocolo de consenso persigue la unificación de las pautas de actuación con el fin de disminuir la incidencia de acontecimientos adversos y maximizar el resultado quirúrgico

REVIEWS AIM: When a phacoemulsification, a filtration surgery or a combined surgery are necessary, questions about the convenience of continuing certain antiglaucomatous drugs could appear. The aim of this review article is to unify criteria that will guide daily clinical practice and including the developing algorithms of action in the preoperative and postoperative periods of filtration surgery and/or cataract surgery. Proposed protocols: In the preoperative period of cataract surgery, the use of non-steroidal anti-inflammatory drugs is at the discretion of the surgeon, with the monodose presentation being recommended. The suspension of prostaglandines a fewdays before the surgery should be considered. Preservative-free drugs ensure a better recovery of the ocular surface (OS) after cataract surgery. Once all modifying factors of the intraocular pressure (IOP) have been removed, baseline IOP should be evaluated again, choosing preservative-free antiglaucomatous drugs when needed. The use of preservative-free ocular antihypertensive drugs and steroids in the preoperative period of glaucoma surgery reduces the risk of surgical failure. The interruption of prostaglandines is recommended. In the postoperative period of glaucoma surgery, steroids are the anti-inflammatory treatment of choice, the preservative-free ones being preferred. When reintroducing antiglaucomatous treatment, preservatives should be avoided to prevent scarring. The appropriate perioperative management of patients with glaucoma is essential to obtain a correct control of IOP, improve the situation of the OS, prevent complications and improve the result of the filtration surgery and cataract surgery. CONCLUSIONS: this protocol aims to unify the different lines of action in order to decrease the incidence of adverse events and maximize the surgical outcome

Human-on-a-chip design strategies and principles for physiologically based pharmacokinetics/pharmacodynamics modeling.

Advances in maintaining multiple human tissues on microfluidic platforms has led to a growing interest in the development of microphysiological systems for drug development studies. Determination of the proper design principles and scaling rules for body-on-a-chip systems is critical for their strategic incorporation into physiologically based pharmacokinetic (PBPK)/pharmacodynamic (PD) model-aided drug development. While the need for a functional design considering organ-organ interactions has been considered, robust design criteria and steps to build such systems have not yet been defined mathematically. In this paper, we first discuss strategies for incorporating body-on-a-chip technology into the current PBPK modeling-based drug discovery to provide a conceptual model. We propose two types of platforms that can be involved in the different stages of PBPK modeling and drug development; these are µOrgans-on-a-chip and µHuman-on-a-chip. Then we establish the design principles for both types of systems and develop parametric design equations that can be used to determine dimensions and operating conditions. In addition, we discuss the availability of the critical parameters required to satisfy the design criteria, consider possible limitations for estimating such parameter values and propose strategies to address such limitations. This paper is intended to be a useful guide to the researchers focused on the design of microphysiological platforms for PBPK/PD based drug discovery.

Estudio descriptivo de los pacientes atendidos en una Unidad de Tabaquismo / No disponible

Objetivo: Conocer las características de los pacientes atendidos en nuestra Unidad de tabaquismo así como los resultados obtenidos en términos de abstinencia. Pacientes y métodos: Se revisan los datos recogidos de los pacientes atendidos durante 7 años en nuestra consulta, y realizamos un estudio descriptivo. Se analizan los datos antropométricos, los relativos al consumo de tabaco, el tratamiento prescrito y la abstinencia en las distintas visitas de seguimiento hasta el año de seguimiento. Análisis estadístico con SPSS 15.0. Resultados: De los 518 pacientes revisados atendidos en nuestra Unidad hay un predominio de mujeres (56,4%), con una edad media de 47,11 años, y un consumo tabáquico de 26,6 cigarrillos al día. El tratamiento pautado con más frecuencia ha sido la terapia sustitutiva con nicotina (TSN). La abstinencia a los 6 meses de seguimiento obtenida fue del 63,8%, y al año del 55,8%. Conclusiones: Atendemos a una población de mediana edad con un consumo de tabaco elevado. El tratamiento farmacológico prescrito con mayor frecuencia es la TSN, logrando la abstinencia a los 6 meses más de la mitad de los pacientes atendidos

Purpose: To know the data of patients attending our Smoking Cessation Unit and their abstinence rate. Patients and methods: Review of the data collected along 7 years and descriptive analysis. We studied anthropometric data, smoking habits, treatment prescribed for smoking cessation and abstinence in each visit until one year of follow-up. Statistical analysis with SPSS 15.0. Results: 518 patients studied, 56.4% females, mean age 47.11 years old, smoked 26.6 cigarettes per day. The pharmacological treatment more frequently prescribed was nicotine replacement therapy (NRT). Six-month abstinence was 63.8% and at one year 55.8%. Conclusions: Our patients are middle-aged and have a high rate of smoking consumption. NRT is the treatment more frequently used, with abstinence in more than half of the patients

Terapias ventilatorias en el manejo del edema agudo de pulmón cardiogénico / Moreno-Zabaleta, R

El edema agudo de pulmón (EAP), fundamentalmente de origen cardiogénico, supone una importante carga asistencial en las urgencias hospitalarias, así como una importante causa de muerte. Junto a un tratamiento médico óptimo, muchas guías recomiendan el uso de CPAP o ventilación mecánica no invasiva. Aunque los meta-análisis publicados hasta ahora muestran suficiente evidencia para recomendar el uso de dispositivos ventilatorios en el edema agudo de pulmón, existe un ensayo clínico que incluye cerca de 1.000 pacientes en el que no se mostró una clara ventaja de la CPAP o la ventilación mecánica no invasiva con respecto a la oxigenoterapia en el tratamiento de estos pacientes. Esto ha generado cierta controversia en el manejo del edema agudo de pulmón con terapias ventilatorias. Como alternativa existen otros dispositivos no mecánicos, como la CPAP de Boussignac o el oxígeno con alto flujo humidificado, que en estudios iniciales parecen tener resultados similares a la CPAP o la ventilación mecánica no invasiva

Acute pulmonary edema (APE), fundamentally of cardiogenic origin, entails a significant care load in the hospital emergency services and is an important cause of death. Together with optimal medical treatment, many guidelines recommend the use of continuous positive airway pressure (CPAP) or non-invasive mechanical ventilation. Although the meta-analyses published up to date show sufficient evidence to recommend the use of ventilatory devices in acute pulmonary edema, there is a clinical trial including approximately 1000 patients in which no clear advantage of the CPAP or non-invasive mechanical ventilation over oxygen therapy in the treatment of these patients was demonstrated. This has generated some controversy regarding the management of acute pulmonary edema with ventilatory therapies. As an alternative, there are other non-mechanical devices such as Boussignac CPAP or high flow humidified oxygen therapy whose results seem to be similar to CPAP or non-invasive mechanical ventilation in the initial studies

Testicular perfusion apparatus.

OBJECTIVES: A reproductive biology laboratory requires to provide reliable instruments for regular use of research workers. To understand an organ and perform scientific study and reach on a conclusion good instruments and techniques are essential. Such a suitable one is not available in case of testicular study. The present study is aimed to clear this lacuna. MATERIALS AND METHODS: A new testicular perfusion apparatus for rabbit and rat is designed and fabricated with glass. The glass blowing instrument available is used. The perfusion fluid preferred is Krebs Ringer bicarbonate buffer (pH 7.4). Quartz wool provided in stem tube filters emboli present in it. Perfusate temperature is maintained to that of testis. A 26-gauge needle is introduced into the spermatic artery to allow entry of perfusate into testis. Method for the collection of perfusate leaving testis is available. This permits to measure the level of an injected substance before its entry through perfusion fluid into the testis, which shall be compared in the fluid after completion of perfusion. RESULTS: Initially few animal testes were used and found the instrument to be functioning well. CONCLUSION: A new simple perfusion apparatus for testes of rabbit and rat is designed. The apparatus made out of glass could be prepared with the help of glass blowing facility and used regularly in any laboratory.

Propriedades farmacológicas da Aloe vera (L. ) Burm. f / Pharmacological activities of Aloe vera (L. ) Burm. f

A Aloe vera (L.) Burm.f. tem sido utilizada há milhares de anos na medicina tradicional para o tratamento de diversos males. O intuito desse trabalho foi o levantamento bibliográfico de artigos que evidenciassem a atividade farmacológica da Aloe vera. A revisão contemplou livros e periódicos nacionais e internacionais indexados nas bases de dados MEDLINE, LILACS e SciElo, nos idiomas português, inglês e espanhol, utilizando as palavras-chave citadas. Após o levantamento bibliográfico, constatou-se que várias atividades biológicas são atribuídas a Aloe vera. Evidências sugerem eficácia no tratamento da psoríase, herpes genital, queimaduras e hiperglicemia. Além disto, também foram demonstradas atividades antineoplásica, antimicrobiana, anti-inflamatória e imunomodulatória por estudos in vitro e in vivo, entretanto, na cicatrização de feridas, os resultados foram conflitantes. No tratamento de dermatite por radiação e em queimaduras solares sua eficácia não foi comprovada e foram relatados casos de hepatite aguda devido ao consumo de preparações orais. Tendo em vista as várias atividades comprovadas e poucos relatos acerca de sua contra indicação, conclui-se que o uso desta espécie corrobora o vasto uso popular.

The Aloe vera (L.) Burm.f. has been used for thousands of years in traditional medicine to treat various ailments. The aim of this study was to carry out a bibliographical review on the pharmacological activity of Aloe vera. This review included books and national and international journals indexed to MEDLINE, LILACS and SciELO, in Portuguese, English and Spanish, using the key words mentioned. After the literature review, we found that several biological activities have been attributed to Aloe vera. Evidence suggests efficacy in the treatment of psoriasis, genital herpes, burns and hyperglycemia. Moreover, antineoplastic, antimicrobial, anti-inflammatory and immunomodulatory activities have also been demonstrated by in vitro and in vivo studies; however, in wound healing, the results were conflicting. In the treatment of radiation dermatitis and sunburn, its efficacy has not been proven, and cases of acute hepatitis from the consumption of oral preparations have been reported. Considering the various proven activities and the few reports about the contraindications of Aloe vera, we conclude that the use of this species confirms its wide popular usage.

Toxins from the Caribbean sea anemone Bunodeopsis globulifera increase cisplatin-induced cytotoxicity of lung adenocarcinoma cells

Lung cancer causes 1.4 million deaths worldwide while non-small-cell lung cancer (NSCLC) represents 80-85% of the cases. Cisplatin is a standard chemotherapy against this type of cancer; however, tumor cell resistance to this drug limits its efficacy. Sea anemones produce compounds with pharmacological activities that may be useful for augmenting cisplatin efficacy. This study aimed to evaluate the pharmacological activities of crude venom (CV) from the sea anemone Bunodeopsis globulifera and four derived fractions (F1, F2, F3 and F4) to test their increase efficiency cisplatin cytotoxicity in human lung adenocarcinoma cells. Results Pre-exposure to CV, F1 and F2 fractions increases cisplatin cytotoxicity in human lung adenocarcinoma cells under specific conditions. Exposure to CV at 50 μgmL-1 induced a reduction of approximately 50% in cell viability, while a similar cytotoxic effect was observed when cell culture was exposed to F1 at 25 μgmL -1 or F2 at 50 μgmL-1. The cell culture exposure to F1 (10 μgmL-1) fraction combined with cisplatine (25 μM) provoked a decrease in MTT reduction until 65.57% while F2 (25 μgmL-1) fraction combined with cisplatin (10 μM) provoked a decrease in MTT reduction of 72.55%. Conclusions The F1 fraction had the greatest effect on the lung adenocarcinoma cell line compared with CV and F2. The combination of antineoplastic drugs and sea anemone toxins might allow a reduction of chemotherapeutic doses and thus mitigate side effects.

Isolated biomolecules of pharmacological interest in hemostasis from Cerastes cerastes venom

Biomolecules from Cerastes cerastes venom have been purified and characterized. Two phospholipases isolated from Cerastes cerastes venom share 51% of homology. CC2-PLA2 exhibits antiplatelet activity that blocks coagulation. CCSV-MPase, a non-hemorrhagic Zn2+-metalloproteinase, significantly reduced the plasmatic fibrinogen level and hydrolyzes only its Bβ chain. Serine proteinases such as RP34, afaâcytin and CC3-SPase hydrolyze the fibrinogen and are respectively α, αβ and αβ fibrinogenases. In deficient human plasma, afaâcytin replaces the missing factors VIII and IX, and activates purified human factor X into factor Xa. It releases serotonin from platelets and directly aggregates human (but not rabbit) blood platelets. RP34 proteinase also had no effect on both human and rabbit blood platelet aggregation. CC3-SPase revealed a pro-coagulant activity. However, the insolubility of the obtained clot indicates that CC3-SPase does not activate factor XIII. In addition, CC3-SPase clotting activity was carried out with human plasmas from volunteer patients deficient in clotting factors. Results showed that CC3-SPase shortens clotting time of plasma deficient in factors II and VII but with weaker clotting than normal plasma. The clotting time of plasma deficient in factor II is similar to that obtained with normal plasma; suggesting that CC3-SPase is able to replace both factors IIa and VII in the coagulation cascade and thus could be involved in the blood clotting process via an extrinsic pathway. These results imply that CC3-SPase and afaâcytin could repair hemostatic abnormalities and may replace some factors missing in pathological deficiency. Afaâcytin also exhibits α fibrinase property similar to a plasmin-like proteinase. Despite its thrombin-like characteristics, afaâcytin is not inhibited by plasmatic thrombin inhibitors. The procoagulant properties of afaâcytin might have potential clinical applications.

Optimizing hollow-fiber-based pharmacokinetic assay via chemical stability study to account for inaccurate simulated drug clearance of rifampicin.

With increasing multidrug resistance coupled to a poor development pipeline, clinicians are exploring antimicrobial combinations to improve treatment outcomes. In vitro hollow-fiber infection model (HFIM) is employed to simulate human in vivo drug clearance and investigate pharmacodynamic synergism of antibiotics. Our overarching aim was to optimize the HFIM-based pharmacokinetic (PK) assay by using rifampicin and polymyxin B as probe drugs. An ultrapressure liquid chromatography tandem mass spectrometry method was validated for the quantification of rifampicin and polymyxin B components. In vitro profiling studies demonstrated that the experimental PK profiles of polymyxin B monotherapy were well correlated with the human population PK data while monotherapy with rifampicin failed to achieve the expected maximum plasma concentration. Chemical stability studies confirmed polymyxin B was stable in broth at 37 °C up to 12 h while rifampicin was unstable under the same conditions over 12 and 80 h. The calculated mean clearance of rifampicin due to chemical degradation was 0.098 ml/min accounting for 12.2 % of its clinical total clearance (CL = 0.8 ml/min) based on population PK data. Our novel finding reinforces the importance to optimize HFIM-based PK assay by performing chemical stability study so as to account for potential discrepancy between experimental and population PK profiles of antimicrobial agents.

[Theory of toxic classification of traditional Chinese medicine and recommendations for revision of China pharmacopeia (volume 1)].

Toxic classification of traditional Chinese medicine, as a contribution of traditional Chinese medicine (TCM) to the recognition of medicinal toxicity and rational use of medicinal materials by Chinese people, is now a great issue related to safe medication, sustainable development and internationalization of Chinese medicine. In this article, the origination and development of toxic classification theory was summarized and analyzed. Because toxic classification is an urgent issue related to TCM industrialization, modernization and internationalization, this article made a systematic analysis on the nature and connotation of toxic classification as well as risk control for TCM industry due to the medicinal toxicity. Based on the toxic studies, this article made some recommendations on toxic classification of Chinese medicinal materials for the revision of China Pharmacopeia (volume 1). From the aspect of scientific research, a new technical guideline for research on toxic classification of Chinese medicine should be formulated based on new biological toxicity test technology such as Microtox and ADME/Tox, because the present classification of acute toxicity of mice/rats can not met the modern development of Chinese medicine any more. The evaluation system and technical SOP of TCM toxic classification should also be established, and they should well balance TCM features, superiority and international requirements. From the aspect of medicine management, list of toxic medicines and their risk classification should be further improved by competent government according to scientific research. In China Pharmacopeia (volume I), such descriptions of strong toxicity, toxicity or mild toxicity should be abandoned when describing medicine nature and flavor. This revision might help promote TCM sustainable development and internationalization, and enhance the competitive capacity of Chinese medicine in both domestic and international market. However, description of strong toxicity, toxicity or mild toxicity might be used when making cautions for the medicine, stating that the description is based on Chinese classic works. In this way, TCM traditional theory might be inherited and features of Chinese medicine maintained and reflected. Besides, modern findings should be added to the cautions, including dose-response relationship, toxic mechanism, and toxic elements. The traditional toxic descriptions and modern findings, as a whole, can make the caution clear and scientific, and then promote safe medication and TCM modernization and internationalization.

Analysis of drug metabolism activities in a miniaturized liver cell bioreactor for use in pharmacological studies.

Based on a hollow fiber perfusion technology with internal oxygenation, a miniaturized bioreactor with a volume of 0.5 mL for in vitro studies was recently developed. Here, the suitability of this novel culture system for pharmacological studies was investigated, focusing on the model drug diclofenac. Primary human liver cells were cultivated in bioreactors and in conventional monolayer cultures in parallel over 10 days. From day 3 on, diclofenac was continuously applied at a therapeutic concentration (6.4 µM) for analysis of its metabolism. In addition, the activity and gene expression of the cytochrome P450 (CYP) isoforms CYP1A2, CYP2B6, CYP2C9, CYP2D6, and CYP3A4 were assessed. Diclofenac was metabolized in bioreactor cultures with an initial conversion rate of 230 ± 57 pmol/h/10(6) cells followed by a period of stable conversion of about 100 pmol/h/10(6) cells. All CYP activities tested were maintained until day 10 of bioreactor culture. The expression of corresponding mRNAs correlated well with the degree of preservation. Immunohistochemical characterization showed the formation of neo-tissue with expression of CYP2C9 and CYP3A4 and the drug transporters breast cancer resistance protein (BCRP) and multidrug resistance protein 2 (MRP2) in the bioreactor. In contrast, monolayer cultures showed a rapid decline of diclofenac conversion and cells had largely lost activity and mRNA expression of the assessed CYP isoforms at the end of the culture period. In conclusion, diclofenac metabolism, CYP activities and gene expression levels were considerably more stable in bioreactor cultures, making the novel bioreactor a useful tool for pharmacological or toxicological investigations requiring a highly physiological in vitro representation of the liver.

[Tools for network pharmacology study: network visualization and network analysis].

Network pharmacology has been one of the academic frontiers of traditional Chinese medicine research. In this paper, we generally showed the work flow of network pharmacology research, and introduced concepts of network visualization and network analysis, as well the commonly-used tools, such as Cytoscape.

Estudios Farmacogenéticos del tratamiento con Antipsicóticos: Estado actual y perspectivas / Pharmacogenetic studies on the antipsychotic treatment. Current status and perspectives

En este trabajo se revisa, a la luz de los conocimientos actuales, la relevancia de los estudios farmacogenéticos en el tratamiento con fármacos antipsicóticos. Se han descrito un gran número de asociaciones entre distintos marcadores genéticos y la respuesta al tratamiento, así como a la aparición de efectos adversos. Sin embargo, no se ha identificado aún ningún biomarcador “estrella” capaz de predecir de forma inequívoca el beneficio clínico de un determinado tratamiento ni su toxicidad. La utilización de marcadores farmacogenéticos individuales se ha demostrado de poca utilidad clínica, por lo que la combinación de la información obtenida del estudio de diversos genes parece una estrategia más prometedora. La inclusión de estudios farmacogenéticos en ensayos clínicos realizados de forma prospectiva incluyendo un elevado número de pacientes podría, sin duda, contribuir de forma significativa al desarrollo de protocolos de medicina personalizada (AU)

Based on present knowledge, in this work we review the importance of the pharmacogenetic tests in the treatment with antipsychotic drugs. Many association shave been reported between different genetic markers and response to treatment as well as to the appearance of adverse reactions. However, up to now, no “prime” biomarker capable of unequivocally predicting the clinical benefits of a specific treatment or its toxicity has been identified. The use of individual pharmacogenetic markers has been demonstrated to have little clinical utility, and therefore the combination of information obtained from the analysis of different genes seems to be a more promising strategy. Inclusion of pharmacogenetic tests in clinical trials conducted prospectively and that include a large number of cases could, undoubtedly, significantly contribute to the development of individualized medicine protocols (AU)