Marco regulatorio de los medicamentos hemoderivados de uso humano / Regulatory framework for human medicinal products

Se hace una revisión integral de la regulación aplicable a los medicamentos hemoderivados, incluyendo la normativa europea así como su transposición a la normativa nacional. Esta regulación compleja y extensa se actualiza y comenta de forma que facilite a los profesionales sanitarios su consulta y aplicación. Los hemoderivados, medicamentos de administración crónica y susceptibles de transmitir enfermedades, tienen gran impacto social y mediático, y su elaboración, de suma importancia en la industria farmacéutica, se basa en la utilización de plasma humano. Se aborda también el debate de la donación de plasma de forma altruista o remunerada y sus consecuencias en la autosuficiencia de un país para la disponibilidad de hemoderivados

A complete review of the regulations applicable to blood derived medicines is made, including European legislation and its transposition into national legislation. This complex and comprehensive regulation is updated and commented to facilitate appropriate guidance to healthcare professionals for its consultation and use. Blood derived products, as chronic medication and susceptible of transmitting diseases, are very important in the pharmaceutical industry, they have a large social and mediatic impact and their manufacturing is based on the use of human plasma as raw material. The discussion on donating plasma altruistically or in remunerated manner and its effect on self-sufficiency for the availability of blood products is also addressed

Detección de alérgenos de cacahuete mediante un sensor de ADN / Detection of peanut allergens by a DNA sensor

En el presente trabajo se propone un genosensor electroquímico para la detección de un segmento de ADN que codifica parte de la proteína alergénica Ara h 2 del cacahuete. El genosensor se basa en un ensayo tipo sándwich, el analito hibrida con dos secuencias de bases, una de ellas inmovilizada sobre un electrodo de oro serigrafiado, formando una monocapa autoensamblada. La optimización del dispositivo se realizó utilizando la metodología de Superficies de Respuesta. La máxima respuesta se encontró para concentraciones de sonda de captura y agente bloqueante, 1 mM y 2,5 mM respectivamente

In the present work an electrochemical genosensor for detecting a DNA segment encoding part of the allergenic protein peanut Ara h 2 is proposed. Genosensors is based on a sandwich assay format, the analyte hybridized with two base sequences, one immobilized onto a screen printed gold electrode, forming a self-assembled monolayer. The optimization of the device was performed using Response Surface Methodology. The maximum response was found to be 1 µM of capture probe concentration and 2,5 mM of blocking agent concentration

Strongly structured DNA sequences as targets for genosensing: sensing phase design and coupling to PCR amplification for a highly specific 33-mer gliadin DNA fragment.

Electrochemical genosensors are becoming cost-effective miniaturizable alternatives to real-time PCR (RT-PCR) methods for the detection of sequence-specific DNA fragments. We report on the rapid detection of PCR amplicons without the need of purification or strand separation. A challenging target sequence for both PCR amplification and electrochemical detection allowed us to address some difficulties associated to hybridization on electrode surfaces. The target was a highly specific oligonucleotide sequence of wheat encoding the most immunogenic peptide of gliadin that triggers the immune response of celiac disease (CD), the 33-mer. With a sandwich assay format and a rational design of the capture and tagged-signaling probes the problems posed by the strong secondary structure of the target and complementary probes were alleviated. Using a binary self-assembled monolayer and enzymatic amplification, a limit of detection of 0.3 nM was obtained. The genosensor did not respond to other gluten-containing cereals such as rye and barley. Coupling to PCR to analyze wheat flour samples required tailoring both the capture and signaling probes. This is the first time that deleterious steric hindrance from long single-stranded regions adjacent to the electrode surface is reported for relatively short amplicons (less than 200 bp). The importance of the location of the recognition site within the DNA sequence is discussed. Since the selected gene fragment contains several repetitions of short sequences, a careful optimization of the PCR conditions had to be performed to circumvent the amplification of non-specific fragments from wheat flour.

[Polio vaccines, eradication and posterradication]. / Vacunas antipoliomieliticas, erradicación y posterradicación.

Vaccination against polio generates herd immunity (both with the attenuated (OPV) and inactivated (IPV) vaccines) and this will allow the eradication of the disease. The OPV vaccine produces 2-4 polio cases per cohort of one million children and therefore IPV is used in countries that can afford its cost (about 15 times more expensive than OPV). In 1988 the World Health Assembly established the polio eradication goal as "interruption of wild poliovirus transmission". If the elimination of wild poliovirus were achieved, the use of OPV will produce annually between 250 and 500 cases of polio in the world. From 1999, it was clear that eradication would require ending of immunization with OPV. On the 25th of January, 2013 it is approved the plan for the eradication and containment of all polioviruses, wild or not, so that no child suffers paralytic poliomyelitis. The most important landmarks include the lack of wild polio cases after 2014, the introduction of at least one dose of IPV in all immunization programs and to cease the type 2 OPV vaccination by the end of 2016 and to stop the use of the oral bivalent vaccine in 2019. To achieve all this, a complex scientific work and economic solidarity will be required.

Vacunas antipoliomieliticas, erradicación y posterradicación / Polio vaccines, eradication and posterradication

La vacunación antipoliomielítica genera inmunidad de grupo (con vacunas atenuadas (VPO) e inactivadas (VPI) y ello permitirá la erradicación de la enfermedad. La VPO produce de 2-4 casos de poliomielitis por cohorte de un millón de niños y por ello los países que pueden hacer frente al coste de la VPI (unas 15 veces más cara) la utilizan. En 1988 la Asamblea de la Organización Mundial de la Salud aprobó el objetivo de la erradicación como “la interrupción de la transmisión de poliovirus salvajes”. Si se conseguía su eliminación, el mantenimiento de la VPO produciría al año entre 250 y 500 casos de poliomielitis en el mundo. Desde 1999 era evidente que la erradicación requeriría la cesación de la vacunación con VPO. El 25 de enero del 2013 se aprobó el plan para la erradicación y la contención de todos los virus de la polio, salvajes o no, para que ningún niño sufra una poliomielitis paralítica. Los hitos más importantes incluyen, la no aparición de casos de polio salvaje tras el año 2014, la introducción de al menos una dosis de VPI en todos los programas de vacunación y que se suspenda la vacunación con VPO tipo 2 al final del 2016 y que en 2019 se pueda cesar de utilizar la vacuna bivalente oral. Para todo ello será preciso un trabajo científico complejo y solidaridad financiera (AU)

Vaccination against polio generates herd immunity (both with the attenuated (OPV) and inactivated (IPV) vaccines) and this will allow the eradication of the disease. The OPV vaccine produces 2-4 polio cases per cohort of one million children and therefore IPV is used in countries that can afford its cost (about 15 times more expensive than OPV). In 1988 the World Health Assembly established the polio eradication goal as “interruption of wild poliovirus transmission”. If the elimination of wild poliovirus were achieved, the use of OPV will produce annually between 250 and 500 cases of polio in the world. From 1999, it was clear that eradication would require ending of immunization with OPV. On the 25th of January, 2013 it is approved the plan for the eradication and containment of all polioviruses, wild or not, so that no child suffers paralytic poliomyelitis. The most important landmarks include the lack of wild polio cases after 2014, the introduction of at least one dose of IPV in all immunization programs and to cease the type 2 OPV vaccination by the end of 2016 and to stop the use of the oral bivalent vaccine in 2019. To achieve all this, a complex scientific work and economic solidarity will be required (AU)

Diseño de un genosensor electroquímico para la detección indirecta de gluten en alimentos / Electrochemical genosensor for indirect wheat gluten detection in food

Se propone un nuevo genosensor electroquímico para la detección de una secuencia específica de ADN que codifica un fragmento inmunogénico de la Alfa-2-gliadina, proteína del gluten de trigo responsable de la celiaquía. El diseño del genosensor se basa en la formación de una monocapa autoensamblada de sonda de captura y un agente bloqueante, mercaptohexanol, sobre electrodos de oro serigrafiados. Se eligió un ensayo tipo sándwich, utilizando una sonda indicadora marcada con biotina y el conjugado estreptavidina-fosfatasa alcalina como molécula de marcaje. La detección del analito se basó en la medida de la corriente de oxidación del 1-naftol, producto formado por la hidrólisis enzimática del 1-naftil-fosfato, mediante voltamperometría de pulso diferencial. Se investigaron y optimizaron los parámetros implicados en la composición de la fase sensora mediante voltametría cíclica, encontrándose como relación óptima sonda de captura: mercaptohexanol 2 microM:9 mM. Con el objetivo de minimizar las adsorciones inespecíficas, se optimizaron las concentraciones de sonda indicadora y conjugado enzima-estreptavidina, especies involucradas en la fase de medida, obteniéndose como valores óptimos 1 microM y 1,075x10-3 g/L respectivamente. El genosensor propuesto presentó una respuesta lineal entre 20 y 250 nM(AU)

A new electrochemical genosensor has been developed for the detection of a specific DNA sequence that encodes for an immunogenic fragment of Alpha-2-gliadin, protein of gluten wheat that plays an important role in celiac disease. The genosensor is based on a mixed self-assembled monolayer consisting on a capture probe and a diluent molecule, mercaptohexanol, both immobilized on screen-printed gold electrodes. A sandwich-type hybridization assay was selected, using a signaling-DNA probe labeled with biotin and streptavidin-alkaline phosphatase as a reporter molecule. Detection of DNA gluten is based on the measurement of the oxidization current of 1-naphthol, product formed by Alpha-naphthyl phosphate enzymatic hydrolysis, by differential pulse voltammetry. Parameters involved in the sensing phase were investigated and optimized by cyclic voltammetry. The optimal capture probe to mercaptohexanol ratio was found to be 2 micreM:9 mM. In order to minimize unspecific adsorptions, both signaling probe and enzyme-streptavidin conjugate concentrations (measurement phase parameters) were optimized (1 micreM and 1.075·10-3 g/L respectively). A linear response from 20 nM to 250 nM is obtained with the proposed genosensor(AU)