Effect of treatment with growth hormone on body composition and metabolic profile of short children born small for gestational age / Efeito do tratamento com hormônio de crescimento na composição corporal e no perfil metabólico de crianças nascidas pequenas para a idade gestacional com baixa estatura

ABSTRACT Objective: To assess the effect of recombinant growth hormone (rGH) on body composition and metabolic profile of prepubertal short children born small for gestational age (SGA) before and after 18 months of treatment. Methods: It is a clinical, non-randomized, and paired study. Children born SGA, with birth weight and/or length <-2 standard deviations (SD) for gestational age and sex, prepubertal, born at full term, of both genders, with the indication for treatment with rGH were included. The intervention was performed with biosynthetic rGH at doses ranging from 0.03 to 0.05 mg/kg/day, administered subcutaneously, once a day at bedtime. Total lean mass (LM) and total fat mass (FM) were carried out using dual-energy X-ray absorptiometry (DXA), and the metabolic profile was assessed for insulin, glycemia, IGF-1 levels and lipid profile. Results: Twelve patients (nine girls, 8.17±2.39 y) were evaluated; three patients dropped out of the study. There was an increase of LM adjusted for length (LMI) (p=0.008), LMI standard deviation score (SDS) adjusted for age and sex (p=0.007), and total LM (p<0.001). The percentage of body fat (BF%) and abdominal fat (AF) remained unaltered in relation to the beginning of treatment. Among the metabolic variables, blood glucose remained within normal levels, and there was a reduction in the number of participants with altered cholesterol (p=0.023). Conclusions: The effect of rGH treatment was higher on LM than in FM, with increased LM adjusted for length and standardized for age and sex. Glycemia remained within the normal limits, and there was a decreased number of children with total cholesterol above the recommended levels.

RESUMO Objetivo: Avaliar o efeito do hormônio de crescimento recombinante (rHC) na composição corporal e no perfil metabólico de crianças pré-púberes com baixa estatura, nascidas pequenas para a idade gestacional (PIG) antes e depois de 18 meses de tratamento. Métodos: Estudo clínico, não randomizado e pareado. Foram incluídas crianças nascidas PIG, com peso e/ou altura ao nascer <-2 desvios padrão (DP) para idade gestacional e sexo, pré-púberes, nascidas a termo, de ambos os sexos, com indicação de tratamento com rGH. A intervenção foi realizada com rGH biossintético com doses variando de 0,03 a 0,05 mg/kg/dia, administrado por via subcutânea, uma vez ao dia ao deitar-se. A massa magra total (LM) e a massa gorda total (MG) foram determinadas por meio de absorciometria de raios X de dupla energia (DXA), e o perfil metabólico foi avaliado com dosagens de insulina, glicemia, IGF-1 e perfil lipídico. Resultados: Doze pacientes (nove meninas, 8,17±2,39 anos) foram avaliados; três pacientes abandonaram o estudo. Houve aumento da LM ajustada para estatura (LMI) (p=0,008), LMI standard deviation scores (SDS) ajustada para idade e sexo (p=0,007) e LM total (p<0,001). O percentual de gordura corporal (GC%) e gordura abdominal (AF) permaneceu inalterado em relação ao início do tratamento. Entre as variáveis metabólicas, a glicemia manteve-se na normalidade, e houve redução do número de participantes com colesterol alterado (p=0,023). Conclusões: O efeito do tratamento com HCr foi maior na MM do que na MG, com o aumento da MM ajustada para altura e padronizada para idade e sexo. A glicemia permaneceu normal e houve redução do número de crianças com colesterol total acima do recomendado.

Otimização da expressão e purificação de proteína recombinante imunossupressora

The discovery of immunosuppressive drugs in the 1960s enabled immense advances in medicine, being used, for example, in transplant patients, with the purpose of treating and/or preventing graft rejection. Studies in the area continue to grow and evolve, in search of updates to the more targeted and specific therapeutic arsenal. At the Immunochemistry Laboratory of the Butantan Institute, a protein in the venom of the Lachesis muta snake was isolated and identified with immunosuppressive properties, named LMVp. The use of recombinant DNA technology has allowed advances in the synthesis of immunobiologicals, being crucial for research into the structure, function and production of therapeutic medicines. With the aim of achieving more effective performance in obtaining LMVp, this work used competent bacteria from the BL21 DE3 STAR PLYSS strain in an attempt to achieve optimization of the expression, purification and concentration processes of the recombinant protein. However, this optimization path did not prove to be efficient, demonstrating that other strategies must be followed to obtain better LMVp performance.

A descoberta das drogas imunossupressoras na década de 60 possibilitou imenso avanço na medicina, sendo utilizado, por exemplo, em pacientes transplantados, com a finalidade de tratamento e/ou prevenção da rejeição de enxertos. Os estudos na área continuam crescendo e evoluindo, em busca de atualizações do arsenal terapêutico mais direcionado e específico. No Laboratório de Imunoquímica do Instituto Butantan, uma proteína no veneno da serpente Lachesis muta foi isolada e identificada com propriedades imunossupressoras, nomeada LMVp. O uso da tecnologia de DNA recombinante permitiu avanços na síntese de imunobiológicos, sendo crucial para pesquisas sobre estrutura, função e produção de medicamentos terapêuticos. Com o objetivo de alcançar um desempenho mais efetivo na obtenção da LMVp, este trabalho utilizou as bactérias competentes da linhagem BL21 DE3 STAR PLYSS na tentativa de alcançar uma otimização dos processos de expressão, purificação e concentração da proteína recombinante. Entretanto, este caminho de otimização não se mostrou eficiente, demonstrando que se deve seguir outras estratégias para se obter melhor rendimento da LMVp.

Expressão de uma acetilcolinesterase isolada do veneno da centopeia Cryptops iheringi

Centipedes are a group of venomous arthropods widely distributed around the world, and due to their well-adapted nature to urban areas, they often cause accidents in humans. Despite their limited medical relevance, centipede envenomation can lead to a range of undesirable symptoms such as burning pain, paresthesia, swelling, and superficial necrosis at the bite site, occasionally progressing to a severe condition in rare cases. A clinical study involving patients treated at the Vital Brazil Hospital of the Butantan Institute showed that most centipede accidents were caused by the Cryptops and Otostigmus genera, accounting for about 90% of cases. Literature reports indicate that centipede venom contains several bioactive compounds, some with potential therapeutic interest. However, despite its significant pharmacological importance, very little is known about the active components of these animals' venom. Therefore, centipede venom could be an excellent source of as-yet-unknown toxins with unexplored biotechnological potential. Cryptops is one of the genera most associated with accidents in humans, and so far, there is no literature study on the toxins of its venom. Through a transcriptomic analysis of the venom gland of Cryotops iheringi conducted by our group, a significant proportion of acetylcholinesterase sequences was observed. As cholinesterases are involved in cholinergic synapses present in the central and peripheral nervous systems, this enzyme becomes an attractive target for the development of new drugs. This project was already underway in our group, where the enzyme was cloned into a bacterial vector. Consequently, this work aims to carry out the expression tests and purification of this molecule.

As centopeias são um grupo de artrópodes venenosos amplamente distribuídos pelo mundo, e por serem animais bem adaptados a áreas urbanas, frequentemente provocam acidentes em humanos. Apesar da pouca relevância médica, o envenenamento por lacraias pode causar uma série de sintomas indesejáveis, como dor ardente, parestesia, edema e necrose superficial no local da picada, podendo, em casos raros, evoluir para um quadro grave. Um estudo clínico com pacientes atendidos no Hospital Vital Brazil do Instituto Butantan, mostrou que a maioria dos acidentes com lacraias foram causados pelos gêneros Cryptops e Otostigmus, representando cerca de 90% dos casos. Têm sido relatados na literatura que o veneno das lacraias contém vários compostos bioativos, alguns com potencial interesse terapêutico, no entanto, apesar da importância farmacológica significante, muito pouco se sabe sobre os componentes ativos do veneno destes animais. Portanto, o veneno de lacraias pode ser uma excelente fonte de toxinas ainda desconhecidas e com potencial biotecnológico inexplorado. Cryptops é um dos gêneros mais associados a acidentes em humanos e, que até o presente momento, não há na literatura nenhum estudo sobre as toxinas de seu veneno. Através de uma análise transcriptômica da glândula de veneno de Cryotops iheringi realizado pelo nosso grupo, foi observada uma grande proporção de sequências de acetilcolinesterase. Uma vez que as colinesterases estão envolvidas em sinapses colinérgicas presentes no sistema nervoso central e periférico, esta enzima torna-se um alvo atraente para o desenvolvimento de novas drogas. Este projeto já estava em desenvolvimento pelo nosso grupo, onde foi realizada a clonagem da enzima em vetor bacteriano. Por conseguinte, este trabalho tem como objetivo realizar os testes de expressão e purificação desta molécula.

The PHH-1V HIPRA vaccine: a new tool in the vaccination strategy against COVID-19 / La vacuna PHH-1V de HIPRA: una nueva herramienta en la estrategia contra la COVID-19

Objectives. Vaccination against SARS-CoV-2 is essential to mitigate the personal, social and global impact of the coro navirus disease (COVID-19) as we move from a pandemic to an endemic phase. Vaccines are now required that offer broad, long-lasting immunological protection from infection in addi tion to protection from severe illness and hospitalisation. Here we present a review of the evidence base for a new COVID-19 vaccine, PHH-1V (Bimervax®; HIPRA HUMAN HEALTH S.L.U), and the results of an expert consensus. Materials and methods. The expert committee consisted of Spanish experts in medicine, family medicine, paediatrics, immunology, microbiology, nursing, and veterinary medicine. Consensus was achieved using a 4-phase process consisting of a face-to-face meeting during which the scientific evidence base was reviewed, an online questionnaire to elicit opinions on the value of PHH-1V, a second face-to-face update meet ing to discuss the evolution of the epidemiological situation, vaccine programmes and the scientific evidence for PHH-1V and a final face-to-face meeting at which consensus was achieved. Results. The experts agreed that PHH-1V constitutes a valuable novel vaccine for the development of vaccination programmes aimed towards protecting the population from SARS-CoV-2 infection and disease. Consensus was based on evidence of broad-spectrum efficacy against established and emerging SARS-CoV-2 variants, a potent immunological re sponse, and a good safety profile. The physicochemical proper ties of the PHH-1V formulation facilitate handling and storage appropriate for global uptake. Conclusions- The physicochemical properties, formula tion, immunogenicity and low reactogenic profile of PHH-1V confirm the appropriateness of this new COVID-19 vaccine (AU)

Objetivos. La vacunación frente al SARS-CoV-2 es funda mental para mitigar el impacto personal, social y global de la enfermedad por coronavirus (COVID-19) a medida que pasa mos de una fase pandémica a una endémica. Actualmente se requieren vacunas que ofrezcan una protección inmunológi ca amplia y duradera contra la infección, además de proteger de la enfermedad grave y la hospitalización. En este artículo se presenta una revisión de la evidencia científica para una nueva vacuna COVID-19, PHH-1V (Bimervax®; HIPRA HUMAN HEALTH S.L.U) y los resultados de un consenso de expertos. Material y métodos. El comité de expertos incluyó ex pertos españoles en medicina, medicina de familia, pediatría, inmunología, microbiología, enfermería y veterinaria. El con senso se logró mediante un proceso de 4 fases que constó de una reunión presencial durante la cual se revisó la evidencia científica, un cuestionario en remoto para obtener opinions sobre el valor de PHH-1V, una segunda reunión presencial de actualización y discusión sobre la evolución de la situación epidemiológica, los programas de vacunas y la evidencia cien tífica para PHH-1V y una última reunión presencial en la que se obtuvo el consenso. Resultados. Los expertos coincidieron en que PHH-1V constituye una vacuna novedosa y valiosa para el desarrollo de programas de vacunación destinados a proteger a la población de la infección y enfermedad por SARS-CoV-2. El consenso se basó en la evidencia del amplio espectro de eficacia contra las variantes establecidas y emergentes del SARS-CoV-2, una res puesta inmunológica potente y un buen perfil de seguridad. Las propiedades fisicoquímicas de la formulación de PHH-1V facilitan la manipulación y el almacenamiento apropiados para la absorción global. Conclusiones. Las propiedades fisicoquímicas, formula ción, inmunogenicidad y bajo perfil reactogénico de PHH-1V confirman la idoneidad de esta nueva vacuna COVID-19 (AU)

Evaluación positiva de medicamentos: febrero, marzo y abril 2023 / Positive assessment of drugs: february, march and april 2023

Se reseñan los medicamentos evaluados y con dictamen positivo por comisión de expertos de la Agencia Española de Medicamentos y Productos Sanitarios o de la Agencia Europea del Medicamento hechos públicos en febrero, marzo y abril de 2023 considerados de mayor interés para los profesionales sanitarios. Se trata de opiniones técnicas positivas que son previas a la autorización y puesta en el mercado del medicamento. (AU)

The drugs assessed by the Spanish Agency for Medicines and Health Products or European Medicines Agency issued in February, March and April 2023, and considered of interest to healthcare professionals, are reviewed. These are positive technical reports prior to the authorization and placing on the market of the product. (AU)

Evaluación positiva de medicamentos: febrero, marzo y abril 2023 / Positive assessment of drugs: february, march and april 2023

Se reseñan los medicamentos evaluados y con dictamen positivo por comisión de expertos de la Agencia Española de Medicamentos y Productos Sanitarios o de la Agencia Europea del Medicamento hechos públicos en febrero, marzo y abril de 2023 considerados de mayor interés para los profesionales sanitarios. Se trata de opiniones técnicas positivas que son previas a la autorización y puesta en el mercado del medicamento. (AU)

The drugs assessed by the Spanish Agency for Medicines and Health Products or European Medicines Agency issued in February, March and April 2023, and considered of interest to healthcare professionals, are reviewed. These are positive technical reports prior to the authorization and placing on the market of the product. (AU)

Successful outcomes of surgery procedures in patient with Glanzmann´s Thrombasthenia

We report a 35-year-old female patient with Glanzmann's thrombasthenia (GT) and severe anemia due to abnormal uterine bleeding secondary to uterine myomatosis. She required several admissions of red blood cells and platelet transfusions. An elective subtotal hysterectomy with salpingo-oophorectomy was proposed and recombinant factor VII was required. Surgical and postoperative outcomes were successful, without surgical complications, bleeding, or hemogram alterations. 4 years later, she required tooth extraction because of periodontal disease and pulp necrosis. In Peru, reports of GT patients requiring major and minor surgical procedures are lacking, given the low disease prevalence and the difficulties related to surgery. The report of these successful cases becomes relevant to continue improving GT management.

Presentamos el caso de una paciente de 35 años con trombastenia de Glanzmann (GT) y anemia severa por sangrado uterino anormal secundario a miomatosis uterina. Requirió varias admisiones de transfusiones de glóbulos rojos y plaquetas. Se propuso histerectomía subtotal electiva con salpingo-ooforectomía y se requirió factor VII recombinante. Los resultados quirúrgicos y postoperatorios fueron exitosos, sin complicaciones quirúrgicas, sangrado ni alteraciones del hemograma. 4 años después, requirió extracción dental por enfermedad periodontal y necrosis pulpar. En Perú faltan reportes de pacientes con GT que requieran procedimientos quirúrgicos mayores y menores, dada la baja prevalencia de la enfermedad y las dificultades relacionadas con la cirugía. El reporte de estos casos de éxito cobra relevancia para seguir mejorando la gestión de GT

Ensayo controlado y aleatorizado del uso de Heberkinasa® en el derrame pleural paraneumónico en niños / Randomized Controlled Trial of the Use of Heberkinase® in Parapneumonic Pleural Effusion in Children

Introducción: El derrame pleural paraneumónico resulta la complicación más frecuente de la neumonía bacteriana, de manejo complejo y muchas veces quirúrgico. No existen publicaciones en Cuba provenientes de ensayos clínicos controlados y aleatorizados ni del uso de la estreptoquinasa recombinante (Heberkinasa®) en el derrame pleural. Objetivo: Evaluar la eficacia y la seguridad de la Heberkinasa® en el tratamiento del derrame pleural paraneumónico complicado complejo y el empiema en niños. Métodos: Ensayo clínico fase III, abierto, aleatorizado (2:1), en grupos paralelos y controlado. Se concluyó la inclusión prevista de 48 niños (1-18 años de edad), que cumplieron los criterios de selección. Los progenitores otorgaron el consentimiento informado. Los pacientes se distribuyeron en dos grupos: I- experimental: terapia estándar y administración intrapleural diaria de 200 000 UI de Heberkinasa® durante 3-5 días y II-control: tratamiento estándar. Las variables principales: necesidad de cirugía y la estadía hospitalaria. Se evaluaron los eventos adversos. Resultados: Ningún paciente del grupo I-experimental requirió cirugía, a diferencia del grupo II-control en el que 37,5 por ciento necesitó cirugía video-toracoscópica, con diferencia altamente significativa. Se redujo la estadía hospitalaria (en cuatro días), las complicaciones intratorácicas y las infecciones asociadas a la asistencia sanitaria en el grupo que recibió Heberkinasa®. No se presentaron eventos adversos graves atribuibles al producto. Conclusiones: La Heberkinasa® en el derrame pleural paraneumónico complicado complejo y empiema resultó eficaz y segura para la evacuación del foco séptico, con reducción de la necesidad de tratamiento quirúrgico, de la estadía hospitalaria y de las complicaciones, sin eventos adversos relacionados con su administración(AU)

Introduction: Paraneumonic pleural effusion is the most frequent complication of bacterial pneumonia, with complex and often surgical management. There are no publications in Cuba from randomized controlled clinical trials or the use of recombinant streptokinase (Heberkinase®) in pleural effusion. Objective: To evaluate the efficacy and safety of Heberkinase® in the treatment of complex complicated parapneumonic pleural effusion and empyema in children. Methods: Phase III, open-label, randomized (2:1), parallel-group, controlled clinical trial. The planned inclusion of 48 children (1-18 years of age), who met the selection criteria, was completed. Parents gave informed consent. The patients were divided into two groups: I-experimental: standard therapy and daily intrapleural administration of 200,000 IU of Heberkinase® for 3-5 days; and II-control: standard treatment. The main variables: need for surgery and hospital stay. Adverse events were evaluated. Results: No patient in group I-experimental required surgery, unlike group II-control in which 37.5 percent required video-assisted thoracoscopic surgery, with a highly significant difference. Hospital stay (to 4 days), intrathoracic complications and infections associated to healthcare in the group that received Heberkinase® was reduced. No serious adverse events attributable to the product occurred. Conclusions: Heberkinase® in complex complicated parapneumonic pleural effusion and empyema was effective and safe for the draining of the septic focus, with reduction of the need for surgical treatment, hospital stay and complications, with no adverse events related to its administration(AU)

First evidence of systemic infection and specific cytotoxic T lymphocyte immune response evoked by oral infection with recombinant Salmonella entérica serovar Albany-Ovalbumin in C57BL/6 mice / Primera evidencia de infección sistémica y respuesta inmune celular citotóxicaespecífica evocada por la infección oral en ratones C57BL/6 con Salmonella entericaserovar Albany-Ovoalbúmina recombinante

Abstract Our group isolated Salmonella enterica serovar Albany from food and feces of wildcaptive carnivores in a zoo from northwestern Mexico. This serovar was also associated with thedeath of an ocelot (Leopardus pardalis) in the same zoo. Another group associated S. Albanywith the death of a human patient. It is due to this zoonotic potential that the in vivo study of thehost-S. Albany relationship is critical. The recombinant S. Albany-Ovalbumin (rSAO) strain wasused to analyze a murine oral infection and its specific cytotoxic T lymphocyte (CTL) response.Our results have shown for the first time that rSAO establishes a systemic infection and evokesepitope-specific lysis with a Th1-like cytokine profile in vivo.

Resumen Salmonella entérica serovar Albany fue aislada por nuestro grupo de investigación de alimentos contaminados y de heces de animales carnívoros en cautiverio en un zoológico del noroeste de México; posteriormente, se logró asociar a este serovar con la muerte de un ocelote (Leopardus pardalis), dentro de este mismo zoológico. Otro grupo de investigación asoció a este serovar con la muerte de un paciente. Es debido a este potencial zoonótico que el estudio in vivo de la relación hospedero-S. Albany es crítico. La cepa recombinante S. Albany-Ovoalbúmina (rSAO) fue utilizada para analizar la infección múrida, al igual que la respuesta inmune celular citotóxica específica. Nuestros resultados demuestran, por primera vez, que rSAO establece una infección sistémica y evoca lisis epítopo-específica con un perfil de citocinas tipo Th1 in vivo.

Tiempo puerta-aguja entre neurólogo presencial y localizado en ictus isquémico tratado con alteplasa. Estudio PRISA / Door-to-needle times for patients with ischaemic stroke treated with alteplase by on-site and off-site on-duty neurologists. PRISA study

Introducción: La guardia de neurología es frecuentemente presencial; es decir, con el neurólogo presente en el centro hospitalario; pero en algunos centros, puede ser localizada o mixta. La fibrinólisis intravenosa (FIV) es uno de los principales tratamientos reperfusores en el ictus isquémico agudo (IIA). El objetivo del siguiente trabajo es comparar el tiempo puerta-aguja (TPA) durante la guardia presencial y la guardia localizada. Material y métodos: Estudio prospectivo, observacional, desde el año 2012 hasta el año 2017 en el que se incluyeron pacientes con IIA y FIV. Se recogieron datos como sexo, edad, hora del inicio de los síntomas, hora de llegada al hospital, hora de la tomografía axial computarizada (TAC), hora de inicio de la FIV. Se consideró guardia de «Presencia» desde las 8:00 hasta las 20:00 y «Localizada» desde las 20:00 hasta las 08:00, divido en 3 grupos: guardia «Presencial», guardia «Localizada» y guardia «Localizada con residente». Resultados: N.° = 138; edad media 69,7 años; mujeres 45,7%. Pacientes en guardia de «Presencia» 96, «Localizada» 42, de los cuales 17 con residente. Ambos grupos presentaban características basales similares. Los TPA de los grupos «Presencia» y «Localizada» fueron 59 y 72 min, respectivamente (p = 0,003). Los TPA de grupos «Presencia», «Localizada» y «Localizada con residente», respectivamente 59, 74 y 68 min (p = 0,001), con significación entre «Presencia» y «Localizada». No se observaron diferentes entre el TPA dependiendo de la franja horaria de mañana, tarde y noche ni entre días laborales y días de fin de semana. Conclusión: La presencia o ausencia del neurólogo en el centro hospitalario en el momento del código ictus, influye en la demora de administración del tratamiento fibrinolítico. La presencia del residente de neurología puede acelerar la realización del proceso. (AU)

Background: Hospital on-call neurology shifts are frequently on-site, but some on-call services may be off-site or mixed. Intravenous tissue plasminogen activator (tPA) is one of the main reperfusion treatments for acute ischaemic stroke (AIS). This study assesses door-to-needle times (DNT) when the neurologist is on-site or off-site. Methods: We performed a prospective, observational study from 2012 to 2017, including patients with AIS and treated with tPA. Data were collected on sex, age, door-to-scan time, scan-to-needle time, and DNT. The on-duty neurologist was on-site from 08:00 to 20:00, and on call but off-site from 20:00 to 8:00. Three groups were formed: on-site, off-site, and off-site with resident present. Results: Our sample included 138 patients. The mean age was 69.7 years, and 45.7% of patients were women. Ninety-six patients were admitted during the on-site shift, 25 during the off-site shift, and 17 during the off-site–resident present shift. Patients admitted during the on-site and off-site shifts presented DNTs of 59 and 72 minutes, respectively (P = .003). DNTs were 59, 74, and 68 minutes (P = .001), respectively, for the on-site, off-site, and off-site–resident present shifts; the difference between DNTs for on-site and off-site shifts was statistically significant. No differences were observed between DNTs according to time of day (morning, afternoon, or night), or between weekdays and weekends. Conclusion: DNT is influenced by whether the on-duty neurologist is on- or off-site at the time of code stroke activation. The presence of a neurology resident can reduce DNT. (AU)

First evidence of systemic infection and specific cytotoxic T lymphocyte immune response evoked by oral infection with recombinant Salmonella enterica serovar Albany-Ovalbumin in C57BL/6 mice.

Our group isolated Salmonella enterica serovar Albany from food and feces of wild captive carnivores in a zoo from northwestern Mexico. This serovar was also associated with the death of an ocelot (Leopardus pardalis) in the same zoo. Another group associated S. Albany with the death of a human patient. It is due to this zoonotic potential that the in vivo study of the host-S. Albany relationship is critical. The recombinant S. Albany-Ovalbumin (rSAO) strain was used to analyze a murine oral infection and its specific cytotoxic T lymphocyte (CTL) response. Our results have shown for the first time that rSAO establishes a systemic infection and evokes epitope-specific lysis with a Th1-like cytokine profile in vivo.

Ética de la edición genética en seres humanos

Propósito/Contexto. Este artículo analiza aspectos éticos de la edición genética en seres humanos. Metodología/Enfoque. Se describe el desarrollo de las principales aplicaciones de la tecnología genética en prevención, diagnóstico y terapéutica de enfermedades genéticas en las últimas décadas, culminando con la edición genética. Resultados/Hallazgos. Se definen los principales aspectos éticos que presenta la edición genética somática y germinal en seres humanos, incluyendo cuestiones de seguridad, especificidad, precisión y certeza. Se critica la edición genética germinal y el concepto de "mejoramiento" humano por vulnerar la autonomía individual, generar cambios genéticos heredables en la progenie y aceptar la falacia del reduccionismo genético de que los rasgos de las personas dependen exclusivamente de la constitución genética, independiente del ambiente. Discusión/Conclusiones/Contribuciones. La edición genética somática puede ser ética si se siguen las normas éticas de la investigación biomédica. Por el contrario, la edición genética germinal no es pertinente ni necesaria para el tratamiento de enfermedades genéticas y presenta graves conflictos éticos, por lo cual, previo a su aplicación es necesario un consenso social por discusiones democráticas, amplias y profundas entre todos los actores sociales involucrados, seguido de mecanismos de gobernanza con regulación robusta por parte del estado, que impidan la vulneración de derechos humanos fundamentales.

Purpose/Context. This article discusses ethical aspects of gene editing in humans. Methodology/Approach. The main applications of genetic technology in the prevention, diagnosis and therapeutics of genetic diseases in recent decades, are described, culminating with genetic editing. Results/Findings. The main ethical aspects of somatic and germline gene editing in humans are discussed, including issues of safety, specificity, precision and certainty. Germline genetic editing and human "enhancement" are criticized for violating individual autonomy, for generating heritable genetic changes in the progeny and for accepting the fallacy of genetic reductionism that people's traits depend exclusively on genetic makeup, independent of the environment. Discussion/Conclusions/Contributions. Somatic gene editing can be ethical if the ethical standards of biomedical research are followed. However, germline genetic editing is not relevant nor necessary for the treatment of genetic diseases and, furthermore, it presents serious ethical conflicts. Therefore, prior to its application, a social consensus is necessary, obtained by democratic, broad and profound discussions among all the social players involved, followed by governance mechanisms with robust regulation by the state, which prevent the violation of fundamental human rights.

Finalidade/Contexto. Este artigo discute aspectos éticos da edição de genes em humanos. Metodologia/Aproximação. Descreve o desenvolvimento das principais aplicações da tecnologia genética na prevenção, diagnóstico e terapia de doenças genéticas nas últimas décadas, culminando com a edição de genes. Resultados/Descobertas. São definidos os principais aspectos éticos da edição de genes somáticos e da linha germinal no ser humano, incluindo questões de segurança, especificidade, precisão e exactidão. A edição genética da Germline e o conceito de "melhoramento" humano são criticados por violarem a autonomia individual, gerando alterações genéticas hereditárias nos descendentes e aceitando a falácia do reducionismo genético de que as características das pessoas dependem exclusivamente da sua constituição genética, independente do ambiente. Discussão/Conclusões/Contribuições. A edição somática de genes pode ser ética se os padrões éticos da investigação biomédica forem seguidos. Pelo contrário, a edição genética na linha germinal não é relevante nem necessária para o tratamento de doenças genéticas e apresenta graves conflitos éticos. Por conseguinte, antes da sua aplicação, é necessário um consenso social através de discussões democráticas, amplas e profundas entre todos os actores sociais envolvidos, seguidas de mecanismos de governação com regulação robusta por parte do Estado, que impeçam a violação dos direitos humanos fundamentais.

Epidural analgesia in pregnant with deficit of factor VII.

The congenital deficit of FVII of coagulation it's an anomaly of genetic transmission autosomal recessive type, it can occur with clinical manifestations like hematomas and spontaneous bleeding or not. The normal levels of FVII it's found between 70%-130% of the laboratory reference value. For unknown reasons there is a poor correlation between levels of FVII and bleeding risk. During pregnancy coagulation can be significantly altered, there is a no clear consensus and a very few information about how to act during labor in a patient with a FVII deficit. The case of a 35-year-old patient with 35 weeks of gestation and congenital deficit of the coagulation FVII (36%) is presented, epidural analgesia is performed during labor previously administering activated recombinant FVII (rFVIIa) without complications (Spinal hematoma, postpartum bleeding, thrombosis).

Analgesia epidural en gestante con déficit del factor VII / Epidural analgesia in pregnant with deficit of factor VII

El déficit congénito del factor VII (FVII) de la coagulación es una anomalía de transmisión genética de tipo autosómico recesivo, puede presentarse con manifestaciones clínicas como hematomas y sangrados espontáneos o no. Los niveles normales del FVII se encuentran entre el 70-130% del valor de referencia del laboratorio. Por razones desconocidas existe una pobre correlación entre los niveles del FVII y el riesgo de sangrado. Durante el embarazo se puede alterar notablemente la coagulación, no existe un consenso claro y hay escasa información sobre cómo actuar durante el trabajo de parto en paciente con déficit del FVII. Se presenta el caso de una mujer de 35 años en la semana 35 de gestación y déficit congénito del FVII (36%), se realiza analgesia peridural durante el parto administrando previamente factor VII recombinante activado (rFVIIa) sin complicaciones (hematoma peridural, sangrado posparto, trombosis).(AU)

The congenital deficit of FVII of coagulation it's an anomaly of genetic transmission autosomal recessive type, it can occur with clinical manifestations like hematomas and spontaneous bleeding or not. The normal levels of FVII it's found between 70%-130% of the laboratory reference value. For unknown reasons there is a poor correlation between levels of FVII and bleeding risk. During pregnancy coagulation can be significantly altered, there is a no clear consensus and a very few information about how to act during labor in a patient with a FVII deficit. The case of a 35-year-old patient with 35 weeks of gestation and congenital deficit of the coagulation FVII (36%) is presented, epidural analgesia is performed during labor previously administering activated recombinant FVII (rFVIIa) without complications (Spinal hematoma, postpartum bleeding, thrombosis).(AU)

Door-to-needle times in patients treated by on-site and off-site on-call neurologists. PRISA study.

BACKGROUND: Hospital on-call neurology shifts are frequently on-site, but some on-call services may be off-site or mixed. Intravenous tissue plasminogen activator (tPA) is one of the main reperfusion treatments for acute ischaemic stroke (AIS). This study assesses door-to-needle times (DNT) when the neurologist is on-site or off-site. METHODS: We performed a prospective, observational study from 2012 to 2017, including patients with AIS and treated with tPA. Data were collected on sex, age, door-to-scan time, scan-to-needle time, and DNT. The on-duty neurologist was on-site from 08:00 to 20:00, and on call but off-site from 20:00 to 8:00. Three groups were formed: on-site, off-site, and off-site with resident present. RESULTS: Our sample included 138 patients. The mean age was 69.7 years, and 45.7% of patients were women. Ninety-six patients were admitted during the on-site shift, 25 during the off-site shift, and 17 during the off-site-resident present shift. Patients admitted during the on-site and off-site shifts presented DNTs of 59 and 72 minutes, respectively (P =  .003). DNTs were 59, 74, and 68 minutes (P =  .001), respectively, for the on-site, off-site, and off-site-resident present shifts; the difference between DNTs for on-site and off-site shifts was statistically significant. No differences were observed between DNTs according to time of day (morning, afternoon, or night), or between weekdays and weekends. CONCLUSION: DNT is influenced by whether the on-duty neurologist is on- or off-site at the time of code stroke activation. The presence of a neurology resident can reduce DNT.

Clonagem, expressão recombinante e validação por microarray de proteínas alimentares potencialmente alergênicas / Cloning, recombinant expression and validation by microarray of potentially allergenic food proteins

A maioria das respostas alérgicas a alimentos é mediada por IgE, que pode ser detectada para fins de diagnóstico da alergia alimentar. No entanto, para isso é necessário que alérgenos purificados estejam disponíveis para a elaboração dos diferentes formatos de ensaio, inclusive por microarray, que se constitui em uma ferramenta bastante útil para análise simultânea, e também para a identificação de reatividade cruzada. A esse respeito, é imprescindível ampliar a plataforma de alérgenos que possam ser empregados para a confecção de microarrays. Atualmente, alguns alimentos que constituem objeto de interesse na clínica em função do número de casos de alergia, e sobre os quais as informações a respeito dos alérgenos são escassas, são: abacaxi, mamão, mandioca e manga. Assim, o objetivo desse trabalho foi clonar, expressar e purificar proteínas potencialmente alergênicas de alimentos de importância regional. Após confirmadas por ensaios imunológicos, essas proteínas foram utilizadas na construção e validação de um microarray através de ensaios com os soros de pacientes alérgicos aos alimentos selecionados. Para atingir esse objetivo, foram selecionadas proteínas potencialmente alergênicas coincidentes, apontadas tanto pela similaridade com espécies taxonomicamente mais próximas, quanto pela técnica 2D Western Blotting acoplada à espectrometria de massas. Dezenove proteínas, sendo 4 de abacaxi, 5 de mamão, 6 de mandioca e 4 de manga, foram expressas em Pichia pastoris, purificadas e impressas em um microarray. Após incubar essas proteínas com os soros dos pacientes alérgicos aos alimentos estudados, 18 proteínas mostraram-se potencialmente alergênicas. Além disso, foi observada reatividade cruzada entre proteínas dos alimentos estudados e também em relação ao látex e outros frutos

The majority of allergic reactions to foods is IgE-mediated, which can be detected for the diagnosis of food allergy. However, purified allergens are necessary to produce different kinds of allergy tests, including microarray, which is a useful tool for simultaneous analysis, as well as for the identification of cross-reactivity. In this respect, it is essential to expand the platform of allergens to include them on microarrays. Nowadays, some foods that are object of interest in the clinical area in Brazil and it is necessary a further evaluation about their potential allergens, since there is a limited information about them, are: pineapple, papaya, cassava and mango. Therefore, the aim of this study was cloning, expressing and purifying potentially allergenic proteins of important Brazilian foods. After confirmed by immunological tests, these proteins were used in microarray production and validation by assays with sera from allergic patients to the selected foods. Achieving this goal, matching potentially allergenic proteins were selected, which were identified by comparison among taxonomically closer species (in silico) and 2D Western Blotting coupled with Mass Spectrometry. Nineteen proteins: 4 from pineapple, 5 from papaya, 6 from cassava and 4 from mango were expressed in Pichia pastoris, purified and printed on a microarray. After incubating those proteins with sera from allergic patients to the selected foods, 18 proteins were detected as potentially allergenic. In addition, cross-reactivity was observed among the proteins from the studied foods, and also regarding to the latex and other fruits

Regulação do processo de migração de células endoteliais (in vitro) por Jararagina-C, uma proteína tipo-disintegrina isolada do veneno de Bothrops jararaca

Jararhagin is a PIII class SVMP with hemorrhagic activity, found in Bothrops jararaca venom. By the process of autolysis, Jararhagin gives rise to Jararhagin-C (JarC), a protein with disintegrin-like and cysteine-rich domains, containing the ECD motif in the disintegrin domain, which selectively recognizes the α2β1 integrin present in platelets and endothelial cells. Several studies have shown the ability of JarC to increase leukocyte adhesion and migration and to promote angiogenesis in experimental models in vivo. However, obtaining JarC from the venom is a limiting factor for the study of this toxin, as it represents about 1% of the crude venom, and expressing this protein in the recombinant form has solved this problem. Escherichia coli was successfully used to express the protein of interest in fusion with the SUMOUlp1 protein. In this work, we used native and recombinant JarC to investigate and compare, in vitro, its action on the migration of endothelial cells CRL-1730 in a Boyden chamber model using type I collagen as substrate and Fetal Bovine Serum (FBS) as cell migration promoting agent. We also verified the gene expression of the IL-8 chemokine by cells incubated with the proteins at 3h, 6h and 24h. Our results showed the effect of both JarC molecules in inhibiting FBS-stimulated cell migration. In the presence of FBS, JarC down regulated the expression of IL-8 at 3h and 6h, however, at 24h there was a slight increase in the expression of this chemokine. Regarding rJarC there was down regulation with 3h, stability with 6h and up regulation with 24h of treatment. However, the behavior of JarC (both in the native and recombinant form) in a culture of cells grown in the absence of FBS was completely different, being possible to observe an up regulation of IL-8 with 3 and 24 hours of stimulation, and a down regulation with 6 hours of stimulation. Our results clearly demonstrate a role for JarC, an ECD-disintegrin, in inhibiting endothelial cell migration. This effect was reproduced by the recombinant protein, confirming that its biological activity is preserved. A protein with an inhibitory action on endothelial cell migration may be a promising tool to treat pathologies that have angiogenesis as a central mechanism, such as age-related macular degeneration or tumor growth.

A Jararagina é uma SVMP de classe PIII com ação hemorrágica encontrada no veneno de Bothrops jararaca. Pelo processo de autólise, a Jararagina dá origem a Jararagina-C (JarC), uma proteína com os domínios tipo-disintegrina e rico em cisteína, contendo o motivo ECD no domínio disintegrina, que reconhece seletivamente a integrina α2β1 presente em plaquetas e células endoteliais. Diversos estudos têm mostrado a capacidade da JarC em aumentar a adesão e migração de leucócitos e promover a angiogênese em modelos experimentais in vivo. Entretanto, a obtenção da JarC a partir do veneno é um fator limitante para o estudo desta toxina, pois ela representa cerca de 1% da massa do veneno seco, e expressar essa proteína de forma recombinante tem solucionado esse problema. A Escherichia coli foi utilizada com sucesso para expressar a proteína de interesse em fusão com a proteína SUMOUlp1. Neste trabalho, utilizamos a JarC nativa e recombinante a fim de investigar e comparar in vitro, sua ação na migração de células endoteliais da linhagem CRL-1730 em modelo de câmara de Boyden usando colágeno tipo I como substrato e Soro Fetal Bovino (SFB) como agente promotor da migração celular. Verificamos também a expressão gênica da quimiocina IL-8 pelas células incubadas com as proteínas em 3h, 6h e 24h. Nossos resultados evidenciaram o efeito de ambas as moléculas de JarC em inibir a migração celular estimulada pelo SFB, sendo a JarC nativa mais eficiente. Na presença de SFB, a JarC regulou negativamente a expressão da IL-8 com 3h e 6h, no entanto, com 24h houve um aumento discreto na expressão desta quimiocina. Com relação a rJarC houve regulação negativa de IL-8 com 3h, estabilidade com 6h e regulação positiva com 24h de tratamento. Porém, o comportamento da JarC (tanto na forma nativa, quanto recombinante) em uma cultura de células cultivadas na ausência de SFB foi completamente diferente, sendo possível observar uma regulação positiva de IL-8 com 3 e 24 horas de estimulo, e uma regulação negativa com 6 horas de estímulo. Nossos resultados demonstram claramente o papel da JarC, uma ECD-disintegrina, inibindo a migração de células endoteliais. Este efeito foi reproduzido pela proteína recombinante, confirmando a sua atividade biológica preservada. Uma proteína com ação inibitória da migração de células endoteliais pode ser uma ferramenta promissora para tratar patologias que tenham a angiogênese como mecanismo central, como por exemplo a degeneração macular relacionada à idade ou ainda o crescimento de tumores.

Administración intrapleural de estreptoquinasa recombinante en tres niños graves con derrame pleural paraneumónico complicado / Intrapleural administration of recombinant streptokinase in three seriously ill children with complicated parapneumonic pleural effusion

Introducción: El derrame pleural paraneumónico es la enfermedad pleural más frecuente de la infancia, el 40-60 por ciento de los casos se presenta como complicación de neumonía adquirida en la comunidad. Ante la aparición de líquido pleural viscoso, con fibrina o tabiques, la fibrinólisis intrapleural aporta beneficios en la resolución de esta grave afección. Objetivo: Describir la evolución clínica e imagenológica de tres niños graves con derrame pleural paraneumónico complicado complejo y empiema. Presentación de casos: Pacientes ingresados en la Unidad de Terapia Intensiva del Hospital Pediátrico Provincial "Dr. Eduardo Agramonte Piña" con el diagnóstico de neumonía adquirida en la comunidad complicada con derrame pleural paraneumónico complicado complejo, tratados con estreptoquinasa recombinante por vía intrapleural; la primera paciente presentó crecimiento de Pseudomona en el cultivo de líquido pleural, patógeno no habitual en las infecciones respiratorias agudas procedentes de la comunidad. El segundo caso, se recibió complicada con shock séptico y el tercer paciente con ecografía torácica que mostró derrame pleural multitabicado, con grandes bolsones y gruesos tabiques. Los tres casos evolucionaron satisfactoriamente, sin necesidad de tratamiento quirúrgico, ni aparición de reacciones adversas atribuibles al fibrinolítico. Conclusiones: La administración intrapleural de la estreptoquinasa recombinante en niños graves con derrame pleural paraneumónico complicado complejo y empiema, resulta un método eficaz y seguro en la evacuación del foco séptico pleural, lo que favorece el control de la infección, sin aparición de complicaciones. Los casos presentados tuvieron evolución satisfactoria y en ninguno se produjo evento adverso relacionado con la administración de la estreptoquinasa recombinante intrapleural(AU)

Introduction: Parapneumonic pleural effusion is the most common pleural disease in childhood; 40-60 percent of cases occur as a complication of community-acquired pneumonia. Given the onset of viscous pleural fluid, with fibrin or septums, intra-pleural fibrinolysis provides benefits to solve this serious condition. Objective: Describe the clinical and imaging evolution of three seriously ill children with complex complicated parapneumonic pleural effusion and empyema. Case presentation: Patients admitted to the Intensive Therapy Unit of "Dr. Eduardo Agramonte Piña" Provincial Pediatric Hospital with the diagnosis of pneumonia acquired in the community worsen with complex complicated parapneumonic pleural effusion, and treated in the intrapleural way with recombinant streptokinase. The first patient showed growth of Pseudomona in the culture of pleural fluid, a pathogen which is not common in acute respiratory infections from the community. The second case was complicated with septic shock; and the third patient had a chest ultrasound that showed multi-sited pleural effusion, with large bags and thick septums. All three cases evolved satisfactorily, without needing surgical treatment, or having adverse reactions attributable to fibrinolytic ones. Conclusions: Intrapleural administration of recombinant streptokinase in seriously ill children with complex complicated parapneumonic pleural effusion and empyema is an effective and safe method in the evacuation of pleural septic focus, which favors infection control, without complications. The cases presented had satisfactory evolution and none of them occurred adversely related to the administration of intrapleural recombinant streptokinase(AU)

Expression of recombinant PreS2-S protein from the hepatitis B virus surface antigen in Pichia pastoris

The aim of this work is the expression of the PreS2-S region of surface antigen of hepatitis B virus (HBV) in yeast Pichia pastoris. A cDNA fragment encoding the Pres2-S protein of HBV was cloned to yeast transfer vectors. Based on cloned new plasmids pPIC3.5-PreS2-S (8707 bp) and pPIC9-PreS2-S (8980 bp) the recombinant strains of P. pastoris producing the PreS2-S region of surface antigen of HBV were obtained. The PAGE electrophoresis and immunoblotting of obtained recombinant PreS2-S protein confirm the molecular weight (34 kDa) and high specificity to the HBV antibodies)AU)

El objetivo de este trabajo es la expresión de la región PreS2-S del antígeno de superficie del virus de la hepatitis B en la levadura Pichia pastoris. Se clonó un fragmento de ADNc que codifica la proteína PreS2-S del VHB en vectores de transferencia de levadura. A partir de los nuevos plásmidos clonados pPIC3.5-PreS2-S (8707 pb) y pPIC9-PreS2-S (8980 pb) se obtuvieron las cepas recombinantes de P. pastoris productoras de la región PreS2-S del antígeno de superficie del VHB. La electroforesis PAGE y la inmunotransferencia de la proteína PreS2-S recombinante obtenida confirman el peso molecular (34 kDa) y la alta especificidad a los anticuerpos contra el VHB(AU)