Tratamientos y bioterapias para COVID-19 / Therapeutics and biotherapies for covid-19

INTRODUCCIÓN: Un nuevo brote de coronavirus surgió en 2019 en Wuhan, China, causando conmoción en el sistema sanitario de todo el mundo; el Comité Internacional de Taxonomía de Virus lo denominó SARS-CoV-2, agente causante de la enfermedad COVID-19.El espectro de gravedad de la enfermedad es muy amplio: la mayoría de los pacientes no presentan gravedad, pero otros pueden desarrollar neumonías, y la insuficiencia respiratoria aguda es la causa más frecuente de mortalidad. Objetivo: analizar y desarrollar las distintas alternativas terapéuticas aportadas por la Biotecnología para tratar los síntomas de aquellos pacientes con COVID-19. Metodología: se realizó una revisión de la bibliografía disponible, a partir de enero de 2020 en PubMed, acerca de los tratamientos que se encuentran aún en ensayos clínicos y aquellos que cuentan con aprobación bajo uso de emergencia para la enfermedad COVID-19. También se realizaron búsquedas a través de Google y Google Académico para publicaciones de organismos de Salud en referencia a políticas de salud establecidas para la terapéutica durante dicha pandemia. Resultados: este trabajo aborda las nuevas alternativas terapéuticas para COVID-19 derivadas de la Biotecnología, que se encuentran tanto en uso como en etapas de ensayos clínicos comprendidos dentro del segmento de los biofármacos y las bioterapias. Se incluye un breve resumen del estatus regulatorio de entidades de salud, el mecanismo de acción de dichas terapias y características generales de cada uno. Se incluyen novedosas bioterapias que se empezaron a implementar para afrontar la pandemia. Conclusiones: la pandemia de coronavirus está poniendo a prueba el sistema sanitario internacional, para brindar soluciones tanto desde el diagnóstico y prevención como para el tratamiento de la población a fin de disminuir la mortalidad. Esto incluyó, obviamente también, al área de la Biotecnología aplicada a la salud, que ha aportado en los tres aspectos mencionados; el presente trabajo se centra en las respuestas de tipo terapéutico que ha brindado y que están comercializadas o en fases clínicas. (AU)

INTRODUCTION: A new coronavirus outbreak emerged in 2019 in Wuhan, China, causing a shock to the healthcare system around the world; the International Committee on Taxonomy of Viruses named it SARS-CoV- 2, the infectious agent of the COVID-19 disease. The spectrum of severity of the disease is very wide, most patients are not serious, but others can develop pneumonia, with acute respiratory failure being the most frequent cause of mortality. Objective: to analyze and develop the different therapeutic alternatives provided by Biotechnology dedicated to Health, to treat the symptoms of those COVID-19 patients who require it, and thus reduce mortality.Methodology: a review of the available literature from January 2020 in PubMed of the treatments that are still in clinical trials and those that have been approved under emergency use for the disease COVID-19 was performed. Searches were also carried out through Google and Google Scholar for publications of Health organizations in reference to health policies established for therapeutics during the mentioned pandemic. Results: this work addresses the new therapeutic alternatives derived from Biotechnology, which are both in use and in stages of clinical trials, to treat patients who developed COVID-19 included within the segment of biopharmaceuticals and biotherapies. A brief summary of the regulatory status of health entities, the mechanism of action of said therapies and general characteristics of each one is included. Innovative biotherapies that began to be implemented to face the pandemic are included. Conclusions: The coronavirus pandemic has driven the international health system to the test, to provide solutions both from the diagnosis, prevention and treatment of the population to reduce the mortality of patients. This obviously also included the area of Biotechnology applied to health, which has contributed in the three aspects mentioned. The present work focuses on the therapeutic responses that it has provided and that are commercialized or in clinical phases. (AU)

Riesgo biológico del blanqueamiento dental interno / Biological risk posed by internal tooth whitening

Introducción: El blanqueamiento dental interno forma parte de la práctica odontológica desde hace varias décadas, pero se han descrito riesgos biológicos de los cuales no existe mucha evidencia. Objetivo: Describir los riesgos biológicos provocados por el blanqueamiento dental interno en dientes no vitales. Métodos: Se realizó una revisión sistemática exploratoria en la base de datos bibliográfica PubMed empleando operadores booleanos y palabras clave que aparecen recogidas en los Descriptores en Ciencias de la Salud. Las variables estudiadas fueron: autor principal, título, año de publicación, tipo de artículo, revista donde se publicaron los artículos y riesgo biológico del blanqueamiento interno descrito. Se filtraron y procesaron los resultados en una base de datos y se analizaron los textos completos para evaluar su calidad y que estos cumplieran los criterios de inclusión y exclusión. Resultados: Fueron incluidos en el estudio 14 artículos, todos por investigaciones originales o revisiones sistemáticas. El 64,3 por ciento de los artículos proyectaba los hallazgos hacia la presencia de reabsorciones cervicales externas posterior al blanqueamiento dental interno. El resto no aportó evidencia alguna de riesgos biológicos luego del tratamiento descrito. Conclusiones: Más de la mitad de los artículos incluidos en el estudio concluyeron que la reabsorción cervical externa es el principal riesgo biológico que trae consigo el tratamiento blanqueador interno de dientes no vitales, apoyados por un factor desencadenante de historial de traumatismos dentarios que propician la aparición de estas afectaciones(AU)

Introduction: Internal tooth whitening has been a component of dental practice for several decades, but biological risks have been described about which not much evidence is available. Objective: Describe the biological risks posed by internal tooth whitening in nonvital teeth. Methods: An exploratory systematic review was conducted in the bibliographic database PubMed using Boolean operators and key words obtained from Health Sciences Descriptors. The variables analyzed were main author, title, year of publication, article type, journal where the articles were published and biological risk of the internal tooth whitening procedure described. Results were filtered and processed in a database and full texts were analyzed for quality and compliance with the inclusion and exclusion criteria. Results: A total 14 articles were included in the study, all of which were either original studies or systematic reviews. In 64.3 percent of the articles, findings had to do with the presence of external cervical resorption after internal tooth whitening. The remaining articles did not contribute any evidence of biological risks after the treatment described. Conclusions: More than half of the articles included in the study concluded that external cervical resorption is the main biological risk posed by internal whitening of nonvital teeth, supported by the triggering factor of a history of dental trauma leading to the occurrence of these disorders(AU)

Efecto de los extractos fitoquímicos en la eliminación del barro dentinario en el tratamiento de conductos radiculares / Effects of phytochemical extracts on smear layer removal in root canal treatment

Introduction: Studies suggested that phytochemical products are considered potential solutions to smear layer removal due to their biological safety in cleaning root canal systems, lower toxicity, lower irritant effect and antimicrobial effect. Objective: The aim of the present study was to perform a systematic review to evaluate smear layer removal of root canal systems by different natural product solutions. Methods: systematic review. Search of the literature was performed on Pubmed, Scopus, Cochrane, Web of Science, Scielo and Google Scholar according the PRISMA protocol. Studies were included if they performed the experiments of smear layer removal on extracted permanent human teeth. Articles published in any language without restriction of year of publication were included in this review. The risk of bias assessment in the included studies was evaluated using the Joanna Briggs Institute Critical Appraisal Checklist for Quasi-Experimental Studies. The search strategy resulted in the retrieval of 6221 publications. After the eligibility criteria application, 8 articles were selected for analysis. Results: It was observed that some natural products solutions showed effects on smear layer removal, especially on coronal third. The phytochemical products such as Salvadora persica, chamomile alcohol extract and grape seed extract demonstrated good potential of smear layer removal, however, its effectiveness and clinical applicability are still unclear. Conclusions: Although the limitations of this study, it is possible to highlight those phytochemical products such as Salvadora persica, chamomile alcohol extract and grape seed extract that demonstrated good potential on SL removal(AU)

Introducción: Los productos fitoquímicos se consideran soluciones potenciales para la eliminación del barro dentinario debido a su seguridad biológica en la limpieza del sistema de conductos radiculares, menor toxicidad, menor efecto irritante y efecto antimicrobiano. Objetivo: Evaluar la eliminación del barro dentinario de los sistemas de conductos radiculares mediante diferentes soluciones de productos naturales. Métodos: La búsqueda de la literatura se realizó en Pubmed, Scopus, Cochrane, Web of Science, SciELO y Google Scholar, según el protocolo PRISMA. Los estudios se incluyeron si realizaron los experimentos de eliminación del barro dentinario en dientes humanos permanentes extraídos. En esta revisión se incluyeron artículos publicados en cualquier idioma, sin restricción de año de publicación. La evaluación del riesgo de sesgo en los estudios incluidos se evaluó mediante la lista de verificación de evaluación crítica del Instituto Joanna Briggs para estudios cuasiexperimentales. La estrategia de búsqueda resultó en la recuperación de 6221 publicaciones. Después de la aplicación de los criterios de elegibilidad, se seleccionaron 8 artículos para su análisis. Resultados: Se observó que algunas soluciones de productos naturales mostraron efectos sobre la eliminación del barro dentinario, especialmente en el tercio coronal. Los productos fitoquímicos como salvadora pérsica, extracto de alcohol de manzanilla y extracto de semilla de uva demostraron un buen potencial de eliminación del barro dentinario; sin embargo, su efectividad y aplicabilidad clínica aún no están claras. Conclusiones: A pesar de las limitaciones de este estudio, es posible destacar aquellos productos fitoquímicos como salvadora pérsica, extracto de alcohol de manzanilla y extracto de semilla de uva demostraron buen potencial de eliminación del barro dentinario(AU)

Espirulina como producto natural con potencialidades para su empleo en pacientes con diabetes mellitus / Spirulina as a natural product with potentials for its use in patients with diabetes mellitus

Introducción: La Spirulina platensis es una cianobacteria planctónica filamentosa, que contiene un espectro natural de mezclas de pigmentos de caroteno, xantofila y ficocianina, con actividad antioxidante y la posibilidad de inducir un mejor control de la glucemia en las personas con diabetes mellitus. Objetivo: Describir los efectos del uso del producto logrado a partir de la bacteria Spirulina platensis en el paciente con diabetes mellitus. Método: Se utilizaron como buscadores de información científica a PubMed, SciELO, Google y Google Académico. Las palabras claves utilizadas fueron: espirulina, Spirulina platensis, Arthrospira platensis, diabetes mellitus y control metabólico. Se evaluaron artículos de revisión, de investigación y páginas web que, en general, tenían menos de 10 años de publicados en idioma español, portugués e inglés, cuyos títulos estaban relacionados con el tema de estudio. Se obtuvieron 70 referencias bibliográficas, de las cuales 49 se citaron en el presente artículo. Conclusiones: La espirulina tiene varios efectos benéficos que permiten su uso como coadyuvante en la prevención y tratamiento de la diabetes mellitus. Es un nutriente con bondades nutraceúticas y funcionales, con potente actividad antioxidante, que incide en un mejor control glucémico y puede ser útil en el manejo de las posibles complicaciones y comorbilidades que acompañan a la diabetes mellitus. Su uso conlleva la posibilidad de algunas reacciones adversas, sobre todo de tipo digestivas, aunque no son frecuentes si se emplean las dosis recomendadas; en general, es considerada un producto seguro(AU)

Introduction: Spirulina platensis is a plankton filamentous cyanobacteria that has a natural spectrum of carotene, xanthophyll and phycocyanin pigments´mix, with antioxidant activity and the possibility of inducing a better control of glycemia in patients with diabetes mellitus. Objective: Describe the effects of the use in patients with diabetes mellitus of a product made from Spirulina platensis bacteria. Method: There were used as scientific information searchers: PubMed, SciELO, Google and Google Scholar. The keywords used were: Spirulina, Spirulina platensis, Arthrospira platensis, diabetes mellitus and metabolic control. There were assessed review articles, research articles and web pages, that in general had less than 10 years of being published in Spanish, Portuguese or English language, and whose titles were related with the studied topic. 70 bibliographic references were collected, and 49 of them were quoted in this article. Conclusions: Spirulina has different beneficial effects that allow its use as coadjuvant agent in the prevention and treatment of diabetes mellitus. It is a nutrient with functional and nutraceutical mildness, with a powerful antioxidant activity which has incidence in a better glycemic control and can be useful in the management of possible complications and comorbidities that accompany diabetes mellitus. Its use entails the possible adverse reactions, mainly digestive ones; although they are not frequent if the recommended doses are used. In general terms, it is considered a safe product(AU)

Risk factors for losing hepatitis B virus surface antibody in patients with HBV surface antigen negative/surface antibody positive serostatus receiving biologic disease-modifying anti-rheumatic drugs: a nested case-control study

Abstract Background: Hepatitis B virus (HBV) reactivation consequent to immunosuppressive therapy is an increasingly prevalent problem with serious clinical implications. Treatment with biologic agents conduces to the loss of protective antibody to HBV surface antigen (anti-HBs), which significantly increases the risk of HBV reactivation. Hence, we investigated the risk factors for losing anti-HBs in patients with rheumatic diseases and HBV surface antigen negative/anti-HBs positive (HBsAg-/anti-HBs+) serostatus during treatment with biologic disease-modifying anti-rheumatic drugs (DMARDs). Methods: Using a nested case-control design, we prospectively enrolled patients with rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis/psoriasis, or juvenile idiopathic arthritis, who were treated with biologic DMARDs at Changhua Christian Hospital, Taiwan, from January 2013 to June 2019 and had HBsAg-/anti-HBs+ serostatus; the analytic sample excluded all patients with HBsAg+ or anti-HBs- serostatus. Anti-HBs titers were monitored 6-monthly and cases were defined as anti-HBs < 10 mIU/ml during follow-up. Cases were matched one- to-all with controls with anti-HBs ≥ 10 mIU/ml on the same ascertainment date and equivalent durations of biologic DMARDs treatment (control patients could be resampled and could also become cases during follow-up). Between-group characteristics were compared and risk factors for anti-HBs loss were investigated by conditional logistic regression analyses. Results: Among 294 eligible patients, 23 cases were matched with 311 controls. The incidence of anti-HBs loss was ∼ 2.7%/person-year during biologic DMARDs treatment. Besides lower baseline anti-HBs titer (risk ratio 0.93, 95% CI 0.89-0.97), cases were significantly more likely than controls to have diabetes mellitus (risk ratio 4.76, 95% CI 1.48-15.30) and chronic kidney disease (risk ratio 14.00, 95% CI 2.22-88.23) in univariate analysis. Risk factors remaining significantly associated with anti-HBs loss in multivariate analysis were lower baseline anti-HBs titer (adjusted risk ratio 0.93, 95% CI 0.88-0.97) and chronic kidney disease (adjusted risk ratio 45.68, 95% CI 2.39-871.5). Conclusions: Besides lower baseline anti-HBs titer, chronic kidney disease also strongly predicts future anti-HBs negativity in patients with HBsAg-/anti-HBs+ serostatus who receive biologic DMARDs to treat rheumatic diseases. Patients with low anti-HBs titer (≤ 100 mIU/ml) and/or chronic kidney disease should be monitored during biologic DMARDs therapy, to enable timely prophylaxis to preempt potential HBV reactivation.

Immunogenicity to biological drugs in psoriasis and psoriatic arthritis

Monoclonal antibodies or fusion proteins, defined as biological drugs, have modified the natural history of numerous immune-mediated disorders, allowing the development of therapies aimed at blocking the pathophysiological pathways of the disease, providing greater efficacy and safety than conventional treatment strategies. Virtually all therapeutic proteins elicit an immune response, producing anti-drug antibodies (ADAs) against hypervariable regions of immunoglobulins. Immunogenicity against biological drugs can alter their pharmacokinetic and pharmacodynamic properties, thereby reducing the efficacy of these drugs. In more severe cases, ADAs can neutralize the therapeutic effects of the drug or cause serious adverse effects, mainly hypersensitivity reactions. The prevalence of ADAs varies widely depending on the type of test used, occurrence of false-negative results, and non-specific binding to the drug, making it difficult to accurately assess their clinical impact. Concomitant use of immunosuppressors efficiently reduces the immunogenicity in a dose-dependent manner, either by decreasing the frequency of detectable ADAs or by delaying their appearance, thereby enhancing the effectiveness of biological therapies. Among the new therapeutic strategies for the management of psoriasis, biological agents have gained increasing importance in recent years as they interrupt key inflammation pathways involved in the physiopathology of the disease. Reports regarding ADA in new biologics are still scarce, but the most recent evidence tends to show little impact on the clinical response to the drug, even with prolonged treatment. It is therefore essential to standardize laboratory tests to determine the presence and titles of ADAs to establish their administration and management guidelines that allow the determination of the real clinical impact of these drugs.

Aplicación del Análisis de Componentes Principales en el proceso de purificación de un biofármaco / Application of Principal Component Analysis in the purification process of a biopharmaceutical

En este trabajo se presenta la aplicación del Análisis de Componentes Principales, mediante el programa THE UNSCRAMBLER versión 8.0, a los datos registrados en un período de 2 años en la etapa de purificación de una planta de producción de Eritropoyetina Humana Recombinante que está basada en varios pasos cromatográficos, de forma similar a los procesos de purificación de proteínas recombinantes que se utilizan como vacunas preventivas o terapéuticas. Se logró reducir dimensionalidad al obtenerse dos componentes principales que explican el 81 por ciento de la varianza de 18 variables originales relacionadas con cuatro pasos cromatográficos. Como resultado se llegó a definir cuáles son las variables que mayor aporte tienen a la variabilidad del proceso en la etapa de purificación, permitiendo extraer información útil para lograr un mayor entendimiento del proceso y enriquecer las estrategias de control en la planta. Dichos resultados corroboraron experiencias prácticas de especialistas de la planta y permitieron dar recomendaciones a considerar en el plan de verificación continuada del proceso como proponer cinco variables como controles de proceso y tener en cuenta que el rendimiento del segundo paso cromatográfico es el más influyente de los rendimientos considerados en la variabilidad(AU)

This paper presents the application of the Principal Component Analysis, using the program THE UNSCRAMBLER version 8.0, to the data recorded during two years in the purification stage of a Recombinant Human Erythropoietin plant that is based on several chromatographic steps, similar to the purification process of recombinant proteins that are used as preventive or therapeutic vaccines. Dimensionality was reduced by obtaining two main components that explain 81 percent of the variance of 18 original variables related to four chromatographic steps. As a result, it was possible to define which variables have the greatest contribution to the variability of the process in the purification stage, allowing to extract useful information to achieve a greater understanding of the process and enrich the control strategies in the plant. These results corroborated practical experiences of plant specialists and allowed for recommendations to be considered in the continuous verification plan of the process, such as proposing three variables as process controls and taking into account that the performance of the second step is the most influential of the performances considered in the variability(AU)

Research progress of natural non-alkaloids with analgesic activity / 中国中药杂志

Pain is a complex, unpleasant feeling and emotional experience associated with actual or potential tissue damage, and manifests itself in certain autonomous psychological and behavioral responses. The commonly used opioid and non-steroidal anti-inflammatory analgesics(NSAIDs) may cause adverse reactions to the kidney, liver, cardiovascular or gastrointestinal system and cause problems of drug abuse. Therefore, it is necessary to study new analgesic drugs with less side effects and significant analgesic effects. A variety of natural products derived from terrestrial plants, microorganisms, marine organisms and fungi have been an important source of clinical medicines and provide an inexhaustible resource for the development and innovation of modern medicines. Therefore, this paper mainly reviews the natural non-alkaloids with analgesic activity in order to provide reference for the research and development of analgesic drugs derived from natural products.

Neuroprotective natural products: promising candidates against neurodegenerative diseases / Productos naturales con propiedades neuroprotectivas: posibles candidatos para el tratamiento de enfermedades neurodegenerativas

Neurodegeneration is a progressive loss of neurons both structurally and functionally causing neuronal cell death ultimately leading to development of various neurodegenerative diseases. Due to poor pharmacokinetic profile of neurotrophins, there still remains a challenge in their neurotrophic therapy where plants, bacteria and fungi, as natural products, could act as promising candidates against various neurological disorders by modulating the neurotrophic activity. Therefore, these natural products that mimic neurotrophins, could develop novel therapeutic approaches to herbal drug that can ameliorate neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and other associated neurological disorders. Taking into account the failure of strategies involving single neurotrophins for the treatment of neurodegenerative diseases, we propose a combination of small molecules of natural products that may work synergistically to restore neuronal functions, minimize side effects and target multiple pathways for a more effective treatment.

La neurodegeneración es una pérdida progresiva de neuronas, tanto estructural como funcional, que causa la muerte neuronal, lo que conduce al desarrollo de diversas enfermedades neurodegenerativas. Debido al pobre perfil farmacocinético de las neurotrofinas, existe un desafío en su terapia neurotrófica donde plantas, bacterias y hongos, como productos naturales, podrían actuar como candidatos contra diversos trastornos neurológicos al modular la actividad neurotrófica. Estos productos naturales que asemejan a las neurotrofinas podrían desarrollar enfoques terapéuticos novedosos como medicamentos a base de hierbas que pueden mejorar enfermedades neurodegenerativas como: Parkinson, Alzheimer y otros trastornos neurológicos asociados. Teniendo en cuenta el fracaso de las estrategias terapéuticas de neurotrofinas para las enfermedades neurodegenerativas, proponemos una combinación de pequeñas moléculas de productos naturales que pueden funcionar sinérgicamente para restaurar las funciones neuronales, minimizar los efectos secundarios y apuntar a múltiples vías para un tratamiento más efectivo.

Incidence of tuberculosis infection in spondyloarthritis patients treated with biological and conventional disease-modifying anti-rheumatic drugs in an endemic area

Introduction: Registries of spondyloarthritis (SpA) patients' follow-up provided evidence that tumor necrosis factor inhibitors (TNFi) increase the incidence of active tuberculosis infection (TB). However, most of these registries are from low burden TB areas. Few studies evaluated the safety of biologic agents in TB endemic areas. This study compares the TB incidence rate (TB IR) in anti-TNF-naïve and anti-TNF-experienced subjects with SpA in a high TB incidence setting.Methods: In this retrospective cohort study, medical records from patients attending a SpA clinic during 13 years (2004 to 2016) in a university hospital were reviewed. The TB IR was calculated and expressed as number of events per 105 patients/year; the incidence rate ratio (IRR) associated with the use of TNFi was calculated.Results: A total of 277 patients, 173 anti-TNF-naïve and 104 anti-TNF-experienced subjects, were evaluated; 35.7% (N = 35) of patients who were prescribed an anti-TNF drug were diagnosed with latent tuberculosis infection (LTBI). Total follow-up time (person-years) was 1667.8 for anti-TNF-naïve and 394.9 for anti-TNF-experienced patients. TB IR (95% CI) was 299.8 (37.4-562.2) for anti-TNF naïve and 1012.9 (25.3-2000.5) for anti-TNF experienced subjects. The IRR associated with the use of TNFi was 10.4 (2.3- 47.9).Conclusions: In this high TB incidence setting, SpA patients exposed to anti-TNF therapy had a higher incidence of TB compared to anti-TNF-naïve subjects, although the TB incidence in the control group was significant.(AU)

Terapias biológicas en embarazo y lactancia / Biological Therapies in Pregnancy and lactation

Las terapias biológicas son una herramienta terapéutica fundamental en cierto grupo de pacientes, sin embargo, su seguridad en el embarazo y lactancia no está aún dilucidada. El objetivo de esta revisión es actualizar los conocimientos y recomendaciones respecto al uso de fármacos biológicos durante el embarazo y lactancia. En relación al embarazo, las recomendaciones actuales sugieren que los anti TNF se podrían continuar o mantener hasta el segundo trimestre del embarazo. La terapia anti CD20 (Rituximab) y otros biológicos se recomiendan suspender en el embarazo. Durante la lactancia, se puede iniciar o continuar cualquier terapia biológica. En la primera mitad del embarazo se pueden utilizar algunos biológicos con relativo buen perfil de seguridad y en la lactancia la evidencia sugiere que no producirían mayor riesgo en el niño. Sería ideal contar con estudios randomizados controlados, pero esto podría generar un dilema ético.

Biological therapies are a fundamental therapeutic tool in a certain group of patients. However, the safety of these drugs during pregnancy and lactation remains unclear. Current recommendations regarding anti TNF use during pregnancy suggest these could be maintained until the second trimester of pregnancy. Anti CD20 therapy (Rituximab) and other biologics should not be used in pregnancy. Patients can safely initiate or continue to use biological therapy during lactation. Some biologicals are relatively safe to use during pregnancy and recent studies show that they can be used during lactation without being a major threat to the child. It would be ideal to have randomized controlled studies, but this could create an ethical dilemma.

Immunological and Morphological substantiation of the use of dental stick with biopharmaceuticals in the complex therapy of inflammatory periodontal diseases

Aim of the study: clinical, immunological and morphological substantiation of the use of a dental stick with biopharmaceuticals in the complex therapy of inflammatory periodontal diseases. Research methods: leukocyte migration test, cytograms, the study of quantitative and qualitative content of spontaneously released mixed saliva, measurement of the total content of protein, the content of secretory immunoglobulin A (sIgA) and lysozyme in it. Clinical improvement of the periodontal tissues condition after treatment by 77% was registered. OHI-S (hygienic index), PI (periodontal disease index), PMA (papillary-marginal-alveolar index) were 7.5, 3.4, and 8.7 times lower, respectively, compared to the pre-treatment group. The depth of periodontal pockets decreased 2.6-3.4 times, the number of sessions per treatment course was reduced to 3-8 visits to the doctor. The amount of mixed saliva (in comparison with the initial data) increased 2.7 times, normal levels were restored and were 1.2 ml higher than control indicators. Protein levels, lysozyme and sIgA concentrations increased and exceeded the pre-treatment level 1.8 times and by 44.5%, respectively. Cytograms data revealed that the number of red blood cells (in one field of view) in the gingiva specimens in inflammatory periodontal diseases (IPD) patients was 2.3 times lower, count of leukocytes with signs of destruction was 13.4 lower and intact leukocyte count was 3.8 times lower. Lymphocyte count was 2 times lower, indicating that the inflammatory process in periodontal tissues was reduced as a re sult of the reduction of the microbial burden: staphylococci and actinomycetes were detected 3 times less often, diplococci ­ 3.5 times less often, filamentous bacteria and streptococci ­ 4 times less often, protists ­ 5 times less often, respectively. The number of cocci microcolonies was 2.7 times higher. The number of fibroblasts (in one field of view) increased threefold, the content of collagen filaments of the normally oriented structure increased 1.4 times, compared to the original data, which indicated the formation of favorable conditions for healing and reparative regeneration processes. The results of the study showed that the use of a dental stick during local IPD therapy is an effective way of correcting changes in clinical and laboratory indicators of local immunity and that dental stick is a promising new dosage form in practical periodontology.

Complexos de cobre com análogos de produtos naturais encontrados em organismos marinhos com atividade antitumoral / Copper complexes with analogues of natural products found in marine organisms with antitumor activity

Neste trabalho foram sintetizados complexos de cobre(II) com derivados imínicos da isatina, incluindo isatinas bromadas semelhantes a compostos encontrados em gastrópodes, a fim de compará-los com o composto já produzido e investigado [Cu(isaepy)], complexo de cobre(II) com base de Schiff feita a partir da isatina e 2-aminoetilpiridina. A isatina é um oxindol produzido em algumas plantas, também encontrado no tecido de mamíferos, com propriedades antitumorais naturais. Isatinas bromadas foram previamente constatadas como mais citotóxicas frente a células tumorais do que a isatina sem substituições. O objetivo principal foi verificar se a presença de bromo nos compostos análogos ao [Cu(isaepy)] levaria a um aumento da atividade antitumoral, assim como maior interação com DNA, alvo usual de metalofármacos. Depois de sintetizados, os compostos foram caracterizados por análise elementar (CHN), espectroscopia no infravermelho, espectroscopia UV/Vis e EPR. Foram feitos testes de citotoxicidade pelo método MTT com células de sarcoma uterino (MES-SA e MES-AS/Dx5, esta última resistente a doxorrubicina), adenocarcinoma cervical (HeLa) e células não cancerosas de fibroblasto humano P4. Adicionalmente, foram feitos testes de interação com DNA por UV/Vis e dicroísmo circular, além de testes de clivagem de DNA plasmidial. De modo geral, foi demonstrado que a simetria tetragonal em torno do cobre, determinada pelo EPR, é importante para a citotoxicidade dos complexos, que dessa forma podem se intercalar ao DNA e impedir sua replicação, por acabar distorcendo a hélice, e pela habilidade de realizarem clivagem oxidativa das fitas. [Cu(isaepy)] e seus análogos bromados demonstraram uma atividade citotóxica muito parecida, assim como grau de interação e clivagem com DNA. Conclui-se que, embora a presença de bromo nos análogos de [Cu(isaepy)] não levem a um aumento de atividade antitumoral, como observado em ligantes correlatos livres, nossos estudos apontam para diferentes fontes naturais (animal ou vegetal) para obtenção de precursores de novos compostos antitumorais

In the present work, copper(II) complexes were synthesized with isatin derived imine ligands, including brominated oxindoles similar to compounds found in gastropods, in order to compare their reactivity with that of [Cu(isaepy)], a Schiff base-copper(II) complex already investigated, obtained with the precursors isatin and 2-aminoethylpyridine. Isatin is a natural oxindole extracted from plants, and also found in mammal tissue, with antitumor properties. Brominated isatins were previously described as much more cytotoxic, towards tumor cells, than unsubstituted isatin. The aim of this work was to verify if the presence of brome in analogue [Cu(isaepy)] compounds would increase their antitumor activity, along with higher DNA interaction, an usual target for metallodrugs. The copper(II) complexes were synthesized and then characterized through elemental analyses (CHN), infrared, UV/Vis and EPR spectroscopies. Cytotoxicity tests were carried out using MTT assay with cells lines MES-SA e MES-SA/Dx5 (uterine sarcome, sensitive and resistent to doxorubicin), HeLa (cervical adenocarcinoma) and non-tumor cells, human fibroblast P4. Additionally, DNA interaction experiments were carried out through UV/Vis spectroscopy and circular dichroism, and at last, DNA cleavage experiments with the studied complexes. In general, it was shown that a tetragonal symmetry around copper, shown by EPR, is very important to the complexes toxicity, since in that way they are able to intercalate DNA, and prevent its replication, as a consequence of double helix distortion, and eventual oxidative cleavage. [Cu(isaepy)] and its brominated analogues demonstrated a very similar cytotoxicity towards cancer cells, as well as quite same level of DNA interaction and cleavage. Although the presence of brome did not increase significantly their antitumor activity, as verified with the free isatin derivatives, our studies pointed to different natural sources to obtain precursors for such new antitumor compounds

PEGylation strategy to the development of analytical and therapeutic proteins / Potencial da PEGuilação para o desenvolvimento de proteínas para fins terapêuticos e analíticos

Protein PEGylation is the covalent bonding of polyethylene glycol (PEG) polymers to amino acid residues of the protein and it is one of the most promising techniques for improving the therapeutic effect of biopharmaceuticals and long-term stability of protein-based biosensors. This chemical modification brings advantages to biopharmaceuticals, such as an increased half-life, enhanced stability, and reduced immunogenicity. Moreover, in the analytical field, PEGylation improves the multiple properties of protein-based biosensors including biocompatibility, thermal and long-term stability, and solubility in organic solvents. However, the use of PEGylated conjugates in the analytical and therapeutic fields has not been widely explored. The limited industrial application of PEGylated bioconjugates can be attributed to the fact that the reaction and separation steps are currently a challenge. The correct selection of the PEGylation reaction design and the purification process are important challenges in the field of bioconjugation. In this sense, the design and optimization of site-specific PEGylation reactions and application of aqueous biphasic systems (ABS) as purification platforms for PEGylated conjugates are the two main objectives of this thesis. Regarding the purification step, the efficient fractionation (i) of the PEGylated conjugates from the native protein and (ii) of the PEGylated conjugates based on their degree of PEGylation was studied. Centrifugal partition chromatography (CPC) was applied as a continuous regime platform based on ABS technology to efficiently purify the PEGylated proteins. The two proteins under study are L-asparaginase, an important biopharmaceutical applied in the treatment of acute lymphoblastic leukemia and cytochrome c, a promising biosensor. The current work developed in this thesis demonstrates the great potential of ABS in the fractionation of PEGylated proteins, under batch and continuous regime. In addition, in situ recovery of the PEGylated products through one-pot bioconjugation and ABS purification was successfully demonstrated for both enzymes studied. Although further research on scale-up is still required, the results presented show the relevance of ABS platforms for the development of separation processes of PEGylated proteins

A PEGuilação de proteínas é a ligação covalente de polímeros de polietilenoglicol (PEG) a resíduos de aminoácidos da proteína e é uma das técnicas mais promissoras para melhorar o efeito terapêutico dos biofármacos e a estabilidade a longo prazo de biossensores proteícos. Esta modificação química traz vantagens aos produtos biofarmacêuticos, como um aumento da meia-vida, maior estabilidade e imunogenicidade reduzida. Além disso, no campo analítico, a PEGuilação melhora as múltiplas propriedades dos biossensores baseados em proteínas, incluindo biocompatibilidade, estabilidade térmica e a longo prazo, e solubilidade em solventes orgânicos. No entanto, o uso de conjugados PEGuilados em campos analíticos e terapêuticos não tem sido amplamente explorado. A aplicação industrial limitada dos bioconjugados PEGuilados pode ser atribuída ao facto de as etapas de reacção e separação serem atualmente um desafio. A seleção correcta do design da reacção de PEGuilação e do processo de purificação são importantes desafios no campo da bioconjugação. Neste sentido, a concepção e otimização de reações de PEGuilação sítio-específicas e aplicação de sistemas aquosos bifásicos (ABS) como plataformas de purificação de conjugados PEGuilados são os dois principais objetivos desta tese. No que concerne à etapa de purificação foi estudado o eficiente fracionamento (i) dos conjugados PEGuilados, da proteína nativa e (ii) dos conjugados PEGuilados baseados no seu grau de PEGuilação. A cromatografia por partição centrífuga (CPC) foi aplicada como uma plataforma de regime contínuo baseada na tecnologia de ABS para purificar eficientemente as proteínas PEGuiladas. As duas proteínas em estudo são a L-asparaginase, importante biofármaco aplicado no tratamento da leucemia linfoblástica aguda e o citocromo c, um potencial biossensor. A partir dos trabalhos desenvolvidos, é possível confirmar o grande potencial dos ABS no fracionamento de proteínas PEGuiladas, em regime contínuo e descontínuo. Além disso, a recuperação in situ dos produtos PEGuilados através da integração em uma única etapa de bioconjugação e purificação por ABS foi comprovada com sucesso para ambas as enzimas estudadas. Embora ainda sejam necessários estudos adicionais sobre a viabilidade destes sistemas em larga escala, os resultados aqui apresentados demonstram a relevância dos ABS para o desenvolvimento de processos de separação de proteínas PEGuiladas

Estudos farmacológicos de novos compostos semissintéticos com potencial terapêutico na doença de chagas / Pharmacological studies of new semi-synthetic compounds with therapeutic potential in Chagas disease

A doença de Chagas afeta mais de 8 milhões de pessoas nos países em desenvolvimento, e dispõe de uma terapia ultrapassada e altamente tóxica. Considerando o Brasil uma das maiores biodiversidades do mundo, compostos de sua flora podem contribuir como novos protótipos farmacêuticos. Neolignanas, isoladas da planta Nectandra leucantha, apresentaram previamente atividade anti-Trypanosoma cruzi e neste trabalho, serviram como base para a semissíntese de uma série de 24 análogos. Estudos in vitro demonstratram que 19 compostos não apresentaram citotoxicidade em células de mamífero até 200 µM. Seis compostos apresentaram atividade contra as formas tripomastigotas (CE50 8 a 76 µM) e 8 compostos contra os amastigotas intracelulares (CE50 7 a 16 µM). Estudos da relação estrutura-atividade biológica (SAR), demonstraram que a presença de pelo menos uma cadeia lateral alílica era importante para a atividade anti-T. cruzi e que o grupo fenol não era essencial. O composto 8 foi eficaz contra as duas formas do parasita e eliminou 100% das formas amastigotas no interior dos macrófagos, sendo escolhido para estudos de mecanismo de ação. O composto 8 afetou rapidamente a mitocôndria do T. cruzi,...(AU)

Chagas disease affects more than 8 million people in developing countries and the available therapy is old and highly toxic. Considering Brazil one of the biggest biodiversity in the world, compounds from the flora can contribute as new pharmaceutical prototypes. Neolignans, isolated from the plant Nectandra leucantha, previously showed anti-Trypanosoma cruzi activity and served as the basis for the semi-synthesis of a series of 24 analogues. In vitro studies demonstrated that 19 compounds caused no cytotoxicity in mammalian cells up to 200 µM. Six compounds showed activity against trypomastigote forms (IC50 8 to 76 µM) and 8 compounds against intracellular amastigotes (IC50 7 to 16 µM). Studies of the structure activity relationships (SAR) showed that the presence of at least one allylic side chain was important for anti-T. cruzi activity and that the phenol group was not essential. Compound 8 was chosen for mechanism of action studies as it was effective against both forms of the parasite and eliminated 100% of the intracellular amastigotes. Compound 8 rapidly affected the mitochondria of T. cruzi, causing an intense depolarization of the mitochondrial membrane potential, with decreasing levels of reactive oxygen species...(AU)

Effects of associations of tannins from Anacardium occidentale and Anadenanthera colubrina with cephalosporin against bovine Staphylococcus aureus isolates / Efeitos da associação de taninos de Anacardium occidentale e Anadenanthera colubrina à cefalosporina sobre Staphylococcus aureus isolados de bovinos

The association of natural compounds isolated from medicinal plants with conventional antibiotics, both with similar mechanisms of action, have become a viable alternative strategy to overcome the problem of drug resistance. This study aimed to evaluate the in vitro antimicrobial activity of tannic substances present in the bark of Anacardium occidentale and Anadenanthera colubrina against samples of Staphylococcus aureus when in combination with cephalexin. These combinations were evaluated by determining the minimum inhibitory concentration (MIC). For this purpose, tannins and cephalexin were serially dissolved in distilled water at concentrations ranging from 0.976 mg/mL to 500 mg/mL and 2 mg/mL to 512 mg/mL, respectively. When combined, the compounds inhibited S. aureus growth forming halos ranging from 0.9 to 46 mm with an MIC of 7.8 mg/mL (tannins) and 4 µg/mL (cephalexin). The resulting effect of the combination of natural and synthetic substances with similar mechanisms of action presented better results than when tested alone. Thus, the conclusion is that both the tannins and cephalexin had their antimicrobial action enhanced when used in combination, enabling the use of lower concentrations while maintaining their antibacterial effect against strains of S. aureus.(AU)

A associação de compostos naturais, isolados de plantas medicinais, com antibióticos convencionais, com mecanismos de ação semelhantes, torna-se uma estratégia alternativa e viável para superar o problema da resistência. Assim, nosso objetivo foi avaliar a atividade antimicrobiana in vitro de substâncias tânicas presentes na casca de Anacardium occidentale e Anadenanthera colubrina associadas à cefalexina, sobre amostras de Staphylococcus aureus. Avaliamos essa associação por meio da determinação da concentração mínima inibitória. Dessa forma, taninos e a cefalexina foram dissolvidos de forma seriada em água destilada em concentrações variando de 0,976 mg/mL a 500 mg/mL e 2 µg/mL a 512 µg/mL, respectivamente. Quando associados, inibiram o crescimento de S. aureus formando halos que variaram de 0,9 a 46 mm com concentração mínima inibitória de 7,8 mg/mL (taninos)/ 4 µg/mL (cefalexina). O efeito resultante da associação de substâncias, natural e sintética, com mecanismos de ação semelhantes, apresentou resultados superiores aos observados quando testados isoladamente. Podemos concluir que os taninos e a cefalexina tiveram sua ação antimicrobiana potencializada quando utilizados em associação, permitindo o uso de uma menor concentração, mantendo seu efeito antibacteriano sobre cepas de S. aureus.(AU)

In vitro antimicrobial activity and alternative control of anthracnose in papaya / Atividade antimicrobiana in vitro e controle alternativo da antracnose em mamão

We evaluated the efficacy of natural products in the control of papaya anthracnose, in vitro and in vivo. The in vitro experiments for evaluation of mycelial growth used a completely randomized 10 × 4 factorial design (treatments × evaluation periods) with eight replicates, with sporulation evaluated at the end of the experiment. The treatments involved the use of aqueous extract at concentrations of 5 and 15% for Syzygium aromaticum (L.) Merr. & Perry (clove), Cinnamomum zeylanicum Breym (cinnamon), and Zingiber officinalis Rox. (ginger); 1 and 3% chitosan; the fungicide Prochloraz at 100 µg.mL-1; and a control (no treatment). For evaluating conidia germination, we used six treatments with five replicates. The treatments included 7.5% of each extract (clove, cinnamon, and ginger), 1.5% chitosan, and 50 µg.mL-1 of Prochloraz. For the in vivo experiment, "Formosa" papaya "Tainung 1" was used in a completely randomized design, with six treatments and four replicates to evaluate the severity of anthracnose caused by Colletotrichum gloeosporioides. The fruits were treated by immersion for 5 min with 15% clove, cinnamon, and ginger extracts, 8% chitosan, and control with distilled water, and immersion for 2 min in Prochloraz (33.75 g a.i./100 L). The treatments with 15% clove extract and 8% chitosan were effective in all evaluations, resulting in a viable alternative to the fungicide Prochloraz. The treatments with ginger extract were less effective and those with cinnamon offered intermediate control.(AU)

Este trabalho teve por objetivo avaliar, in vitro e in vivo, a eficácia de produtos naturais no controle da antracnose do mamão. Os experimentos in vitro foram instalados em delineamento inteiramente casualizado, sendo para a avaliação de crescimento micelial considerado o esquema fatorial 10 × 4 (tratamentos x períodos de avaliação) com oito repetições, com a esporulação avaliada ao final do experimento. Os tratamentos utilizados foram: extrato aquoso nas concentrações de 5 e 15% de Syzygium aromaticum (L.) Merr. & Perry (cravo-da-índia), Cinnamomum zeylanicum Breym (canela) e Zingiber officinalis Rox (gengibre), quitosana 1 e 3%, fungicida Procloraz a 100 µg.mL-1 e testemunha (sem tratamento). Para a avaliação da germinação de conídios, foram considerados seis tratamentos com cinco repetições. Foram utilizados os mesmos tratamentos, porém, nas concentrações de 7,5% de cada extrato (cravo-da-índia, canela e gengibre), 1,5% de quitosana, e 50 µg.mL-1 de Procloraz. Para o experimento in vivo, foram utilizados mamões "Formosa" "Tainung 1", em delineamento inteiramente casualizado, sendo seis tratamentos com quatro repetições para avaliação de severidade de antracnose causada por Colletotrichum gloeosporioides. Os frutos foram tratados por imersão durante 5 minutos com extratos de cravo-da-índia, canela e gengibre a 15%, quitosana a 8% e testemunha com água destilada, e imersão por 2 minutos em fungicida Procloraz (33,75 g i.a./100 L). Os tratamentos com extrato de cravo-da-índia a 15% e quitosana a 8% foram eficazes em todas as avaliações, sendo uma alternativa viável ao fungicida Procloraz. Os tratamentos com extrato de gengibre foram menos eficientes, com controle intermediário para o extrato de canela.(AU)

Determinación de la eficacia de los desinfectantes empleados en las áreas asépticas de un centro productor de biofarmacéuticos / Determination of the efficacy of the disinfectants employed in the aseptic areas at a biopharmaceuticals production center

En la industria biofarmacéutica, la desinfección se ha hecho parte fundamental de las actividades para controlar el nivel de microorganismos contaminantes que puedan llegar a comprometer la calidad del producto final. El objetivo de este trabajo fue evaluar la eficacia de los desinfectantes AniosSpecial DJP SF, Aniosurf Premium, Bacteranios SF, Aniospray 29, Aseptanios AD y Surfanios utilizados en la limpieza y desinfección del área aséptica de la Planta de Productos Parenterales 2 del Centro Nacional de Biopreparados (Mayabeque, Cuba). Se comprobó la eficacia con pruebas de desafío en superficies y en condiciones prácticas de uso. Se comparó estadísticamente el número de microorganismos aislados y la microbiota presente en el área antes y después del uso de los desinfectantes. Todos los desinfectantes cumplieron con los requisitos establecidos. Por lo tanto, se demostró su eficacia para realizar la limpieza de las áreas asépticas(AU)

In the biopharmaceutical industry, disinfection has become a fundamental part of the activities to control the level of contaminants microorganisms that could compromise the quality of the final product. The objective of this work was to evaluate the efficacy of the disinfectants Anios Special DJP SF, Aniosurf Premium, Bacteranios SF, Aniospray 29, Aseptanios AD and Surfanios for the cleaning and disinfection of the aseptic area at the Parenterals Products Plant 2 (PPP2) at the National Center for Bioproducts (Mayabeque, Cuba). The efficacy was checked with challenge tests in surfaces, in practical usage conditions. The number of isolated microorganisms and the microbiota present before and after use was compared statistically. All the disinfectants complied with the established requirements. Therefore, the efficacy of the disinfectants was demonstrated to perform the cleaning and disinfection of the aseptic areas(AU)

Reevaluation of the Strain Pressure Theory: Toward a Better Understanding the Biology of Tooth Movement / Reevaluación de la Teoría Presión - Tensión: Hacia una Mejor Comprensión de la Biología del Movimiento Dental

The aim of this report was to analyze the majority of publications referred to orthodontic movement Biological basis, principally strain- pressure theory taking account contemporaneous methodological study designs. The literature published in Pubmed from 1967 to 2014 was reviewed, in addition to well-known scientific reports that were not classified under this database like classical manuscripts since 1839. The comment items are: Limitations of animal models, Histological slices, what to watch for, Rats as an experimental model in orthodontics, Strain­pressure, the force used in these studies that fundament this theory, Reevaluation studies about strain-pressure theory in biology movement using contemporaneous models, Up to date concepts, Theory reevaluation the first step to understanding new acceleration concepts and Movement biology research, present and future. After analyzing those items we concluded: A 3D perspective is essential for a complete understanding of events in tooth movement involves two interrelated processes: the bending of alveolar bone and remodeling of the periodontal tissues. It's important to start, and acknowledge that the PDL cannot be divided into two different areas that react independently to an applied force. It´s a continuous periodontal compartment in OTM, rather than a pressure side and a tension side. When an orthodontic force is applied there are two simultaneous effects that correlate. Alveolar bone flexion and remodeling of the periodontal tissues. Alveolar Bone deformation compromising the main structures (PDL, bone and cement), compression and tension can coexist in different directions. The magnitudes of compression and tension are typically different in different directions. As the majority of research and studies indicate that fundamental movement biology are made on animal models, it is important to know that their results and conclusions, under the actual evidenced based practice guidelines do not allow to make clinical decisions even if they are included on systematic reviews.

El objetivo de esta revisión es analizar la mayoría de las publicaciones relacionadas a la fundamentación sobre el movimiento de ortodoncia, principalmente la teoría presión-tensión, teniendo en cuenta los diseños de estudios metodológicos contemporáneos. Se revisó la literatura publicada en la base de datos de MEDLINE, 1967-2013 , además de los informes conocidos que no fueron clasificados en esta base de datos como manuscritos clásicos desde 1839. Los temas principales en los que se divide el artículo son: Limitaciones de los modelos animales, cortes histológicos, las ratas como modelo experimental en ortodoncia, la fuerza utilizada en los estudios que fundamentan esta teoría, reevaluación sobre los estudios de la teoría presión ­tensión en modelos contemporáneos, conceptos actuales en biología del movimiento, reevaluación de la teoría: el primer paso para entender los nuevos métodos en aceleración del movimiento, conceptos e investigaciones en biología del movimiento: presente y futuro. Una perspectiva 3D es esencial para una comprensión completa de los acontecimientos. El movimiento dental implica dos procesos interrelacionados: la curvatura del hueso alveolar y la remodelación de los tejidos periodontales. Es importante empezar, y reconocer que el PDL no puede ser dividido en dos áreas diferentes que reaccionan de forma independiente a una fuerza aplicada. Está en un compartimiento continuo periodontal , en lugar de un lado de presión y un lado de tensión. Al aplicar una fuerza de ortodoncia hay dos efectos simultáneos: la flexión del hueso Alveolar y la remodelación de los tejidos periodontales. La deformación del hueso compromete las estructuras principales (PDL, hueso y cemento) y la compresión y la tensión pueden coexistir en diferentes direcciones.