30 research outputs found
Cine y Biología: Una experiencia docente realizada por los estudiantes
El artículo narra una experiencia de un ciclo de cine organizada por los propios estudiantes susceptible de ser reconocida para obtener créditos de libre elección. La actividad fue realizada durante el curso 2009-10 en los estudios de Biología de la Universidad Pompeu Fabra (UPF) de Barcelona.La organización de la actividad y la evaluación de los alumnos matriculados fueron llevadas a cabo por los estudiantes organizadores bajo la supervisión del decano de la facultad.La experiencia fue evaluada positivamente por el alto número de matriculados, por la participación de los mismos y por el resultado de la evaluación de las memorias que tenían que realizar los matriculados. La actividad podría considerarse como un buen ejemplo de implicación de los propios estudiantes en su formación, tanto técnica como humana.</p
Analysis of factors affecting the variability of a quantitative suspension bead array assay measuring IgG to multiple Plasmodium antigens
Reducing variability of quantitative suspension array assays is
key for multi-center and large sero-epidemiological studies. To
maximize precision and robustness of an in-house IgG multiplex
assay, we analyzed the effect of several conditions on
variability to find the best combination. The following assay
conditions were studied through a fractional factorial design:
antigen-bead coupling (stock vs. several), sample predilution
(stock vs. daily), temperature of incubation of sample with
antigen-bead (22°C vs. 37°C), plate washing (manual vs.
automatic) and operator expertise (expert vs. apprentice). IgG
levels against seven P. falciparum antigens with heterogeneous
immunogenicities were measured in test samples, in a positive
control and in blanks. We assessed the variability and MFI
quantification range associated to each combination of
conditions, and their interactions, and evaluated the minimum
number of samples and blank replicates to achieve good
replicability. Results showed that antigen immunogenicity and
sample seroreactivity defined the optimal dilution to assess the
effect of assay conditions on variability. We found that a
unique antigen-bead coupling, samples prediluted daily,
incubation at 22°C, and automatic washing, had lower
variability. However, variability increased when performing
several couplings and incubating at 22°C vs. 37°C. In addition,
no effect of temperature was seen with a unique coupling. The
expertise of the operator had no effect on assay variability but
reduced the MFI quantification range. Finally, differences
between sample replicates were minimal, and two blanks were
sufficient to capture assay variability, as suggested by the
constant Intraclass Correlation Coefficient of three and two
blanks. To conclude, a single coupling was the variable that
most consistently reduced assay variability, being clearly
advisable. In addition, we suggest having more sample dilutions
instead of replicates to increase the likelihood of sample MFIs
falling in the linear part of the antigen-specific curve, thus
increasing precision
Host age and expression of genes involved in red blood cell invasion in Plasmodium falciparum field isolates
Plasmodium falciparum proteins involved in erythrocyte invasion
are main targets of acquired immunity and important vaccine
candidates. We hypothesized that anti-parasite immunity acquired
upon exposure would limit invasion-related gene (IRG) expression
and affect the clinical impact of the infection. 11 IRG
transcript levels were measured in P. falciparum isolates by
RT-PCR, and IgG/IgM against invasion ligands by Luminex(R), in
50 Mozambican adults, 25 children with severe malaria (SM) and
25 with uncomplicated malaria (UM). IRG expression differences
among groups and associations between IRG expression and
clinical/immunologic parameters were assessed. IRG expression
diversity was higher in parasites infecting children than adults
(p = 0.022). eba140 and ptramp expression decreased with age (p
= 0.003 and 0.007, respectively) whereas p41 expression
increased (p = 0.022). pfrh5 reduction in expression was abrupt
early in life. Parasite density decreased with increasing pfrh5
expression (p < 0.001) and, only in children, parasite
density increased with p41 expression (p = 0.007), and decreased
with eba175 (p = 0.013). Antibody responses and IRG expression
were not associated. In conclusion, IRG expression is associated
with age and parasite density, but not with specific antibody
responses in the acute phase of infection. Our results confirm
the importance of multi-antigen vaccines development to avoid
parasite immune escape when tested in malaria-exposed
individuals
Cytoadhesion to gC1qR through Plasmodium falciparum Erythrocyte Membrane Protein 1 in Severe Malaria
Cytoadhesion of Plasmodium falciparum infected erythrocytes to
gC1qR has been associated with severe malaria, but the parasite
ligand involved is currently unknown. To assess if binding to
gC1qR is mediated through the P. falciparum erythrocyte membrane
protein 1 (PfEMP1) family, we analyzed by static binding assays
and qPCR the cytoadhesion and var gene transcriptional profile
of 86 P. falciparum isolates from Mozambican children with
severe and uncomplicated malaria, as well as of a P. falciparum
3D7 line selected for binding to gC1qR (Pf3D7gC1qR). Transcript
levels of DC8 correlated positively with cytoadhesion to gC1qR
(rho = 0.287, P = 0.007), were higher in isolates from children
with severe anemia than with uncomplicated malaria, as well as
in isolates from Europeans presenting a first episode of malaria
(n = 21) than Mozambican adults (n = 25), and were associated
with an increased IgG recognition of infected erythrocytes by
flow cytometry. Pf3D7gC1qR overexpressed the DC8 type PFD0020c
(5.3-fold transcript levels relative to Seryl-tRNA-synthetase
gene) compared to the unselected line (0.001-fold). DBLbeta12
from PFD0020c bound to gC1qR in ELISA-based binding assays and
polyclonal antibodies against this domain were able to inhibit
binding to gC1qR of Pf3D7gC1qR and four Mozambican P. falciparum
isolates by 50%. Our results show that DC8-type PfEMP1s mediate
binding to gC1qR through conserved surface epitopes in DBLbeta12
domain which can be inhibited by strain-transcending functional
antibodies. This study supports a key role for gC1qR in
malaria-associated endovascular pathogenesis and suggests the
feasibility of designing interventions against severe malaria
targeting this specific interaction
Identifying Immune Correlates of Protection Against Plasmodium falciparum Through a Novel Approach to Account for Heterogeneity in Malaria Exposure
Background: A main criterion to identify malaria vaccine
candidates is the proof that acquired immunity against them is
associated with protection from disease. The age of the studied
individuals, heterogeneous malaria exposure, and assumption of
the maintenance of a baseline immune response can confound these
associations. Methods: Immunoglobulin G/immunoglobulin M (IgG/
IgM) levels were measured by Luminex(R) in Mozambican children
monitored for clinical malaria from birth until 3 years of age,
together with functional antibodies. Studied candidates were
pre-erythrocytic and erythrocytic antigens, including
EBAs/PfRhs, MSPs, DBLs, and novel antigens merely or not
previously studied in malaria-exposed populations. Cox
regression models were estimated at 9 and 24 months of age,
accounting for heterogeneous malaria exposure or limiting
follow-up according to the antibody's decay. Results:
Associations of antibody responses with higher clinical malaria
risk were avoided when accounting for heterogeneous malaria
exposure or when limiting the follow-up time in the analyses.
Associations with reduced risk of clinical malaria were found
only at 24 months old, but not younger children, for IgG breadth
and levels of IgG targeting EBA140III-V, CyRPA, DBL5epsilon and
DBL3x, together with C1q-fixation activity by antibodies
targeting MSP119. Conclusions: Malaria protection correlates
were identified, only in children aged 24 months old when
accounting for heterogeneous malaria exposure. These results
highlight the relevance of considering age and malaria exposure,
as well as the importance of not assuming the maintenance of a
baseline immune response throughout the follow-up. Results may
be misleading if these factors are not considered
Duración del proceso de financiación en España de los fármacos innovadores aprobados por la Agencia Europea del Medicamento: 2008-2013
Duración del proceso de financiación en España de los fármacos innovadores aprobados por la Agencia Europea del Medicamento: 2008-2013
Background: In Spain, the decision of Price and Reimbursement (P&R) of a new drug must be taken between 180-270 days. The objective of this study was to assess the reimbursement timing in Spain for innovative drugs approved by the European Medicines Agency (EMA) between January 2008 and December 2013 and to explore the potential impact of drug's price on this time. Methods: Drugs approved were extracted from EMA's website, authorization dates in Spain from the Spanish Agency (AEMPS) and, P&R dates and prices from Nomenclátor and BotPlus. Depending on days from approval to reimbursement, drugs were quick (270). Depending on posology: chronic or acute. Depending on dispensing conditions: retail or hospital drugs. It was calculated: median, maximum, minimum, first, and third quartiles of time until reimbursement. Results: 431 drugs were approved by EMA; 285 were innovative, from them 147 were approved by the AEMPS and reimbursed: 103 chronic and 44 acute. Median price/day was €2.44 for chronic and €21 for acute. From 2008-2011, 80% of drugs were reimbursed, in 2012 21% and in 2013 17%. Time from approval to reimbursement move from 230 days in 2009 to 431 days in 2013. From the 139 drugs with reimbursement date 33 were quick, 44 on time and 62 delayed. Conclusions: The median time from approval by the EMA of innovative drugs since the reimbursement in Spain in 2013 is double that of 2008. The main driver of delays in the process of P&R seems to be the budget impact of the drug instead of its unit price.Fundamentos: En España la decisión de precio y financiación (PyF) de un nuevo medicamento debe tomarse entre 180 y 270 días. El objetivo de este estudio fue valorar el tiempo hasta la financiación en España de los medicamentos innovadores aprobados por la Agencia Europea de Medicamentos (EMA) entre enero 2008 y diciembre 2013 y explorar el impacto potencial del precio del fármaco sobre este tiempo. Métodos: Los medicamentos aprobados se obtuvieron de la web de la EMA, las fechas de autorización de la Agencia Española del Medicamento (AEMPS) y las fechas del PyF y los precios del Nomenclátor y BotPlus. Según los días desde la aprobación hasta la financiación se clasificó a los medicamentos en rápidos (270). Según la duración del tratamiento en crónicos o agudos. Y según las condiciones de dispensación: de farmacia u hospital. Del tiempo transcurrido hasta la financiación, se calculó la mediana, el máximo, el mínimo así como el primer y tercer cuartiles. Resultados: Durante el período de estudio fueron aprobados por la EMA 431 medicamentos, de los cuales 285 eran innovadores. De estos 147 fueron aprobados por la AEMPS y financiados: 103 para tratamientos crónicos y 44 para agudos. La mediana del precio/día fue de 2,44 € para crónicos y 21 € para agudos. De 2008-2011 fueron financiados el 80% , en 2012 el 21% y en 2013 el 17%. El tiempo hasta la financiación pasó de 230 días en 2009 a 431 en 2013. Los 139 medicamentos con fecha de financiación fueron: 33 rápidos, 44 en plazo y 62 con demora. Conclusiones: La mediana del tiempo desde la aprobación por la EMA de los medicamentos innovadores hasta su financiación en España en 2013 es el doble que en 2008. El motor principal de los retrasos en el proceso de PyF parece ser el impacto presupuestario del fármaco más que su precio unitario
Cinema and Biology: A teaching experience carried out by students
[ES] El artículo narra una experiencia de un ciclo de cine organizada por los propios estudiantes susceptible
de ser reconocida para obtener créditos de libre elección. La actividad fue realizada durante el curso
2009-10 en los estudios de Biología de la Universidad Pompeu Fabra (UPF) de Barcelona.
La organización de la actividad y la evaluación de los alumnos matriculados fueron llevadas a cabo por
los estudiantes organizadores bajo la supervisión del decano de la facultad.
La experiencia fue evaluada positivamente por el alto número de matriculados, por la participación de
los mismos y por el resultado de la evaluación de las memorias que tenían que realizar los matriculados.
La actividad podría considerarse como un buen ejemplo de implicación de los propios estudiantes en su
formación, tanto técnica como humana. [EN] The paper recounts the experience of a cycle of film showings organized by the students themselves,
which was also recognized as being an activity whereby elective credits could be obtained. The activity
was carried out during the 2009-10 academic year in the Biology degree at the Pompeu Fabra University
(UPF) in Barcelona.
The organization of the activity and the assessment of the students enrolled were undertaken by the
Student Organizers under the supervision of the Dean of the Faculty of Health and Life Sciences.
The experience was deemed to be highly successful for the following reasons: the number of students
enrolled on it, the notable participation and the result of the assessment of the reports handed in by the
students. The activity could be considered a good example of students’ involvement in their own training,
both technical and human
Protein Kinase C epsilon-Calcineurin Cosignaling Downstream of Toll-Like Receptor 4 Downregulates Fibrosis and Induces Wound Healing Gene Expression in Cardiac Myofibroblasts
The pathways which regulate resolution of inflammation and contribute to positive remodeling of the myocardium following injury are poorly understood. Here we show that protein kinase C epsilon (PKCε) cooperates with the phosphatase calcineurin (CN) to potentiate induction of cardioprotective gene expression while suppressing expression of fibrosis markers. This was achieved by detailed analysis of the regulation of cyclooxygenase 2 (COX-2) expression as a marker gene and by using gene expression profiling to identify genes regulated by coexpression of CN-Aα/PKCε in adult rat cardiac myofibroblasts (ARVFs) on a larger scale. GeneChip analysis of CN-Aα/PKCε-coexpressing ARVFs showed that COX-2 provides a signature for wound healing and is associated with downregulation of fibrosis markers, including connective tissue growth factor (CTGF), fibronectin, and collagens Col1a1, Col3a1, Col6a3, Col11a1, Col12a1, and Col14a1, with concomitant upregulation of cardioprotection markers, including COX-2 itself, lipocalin 2 (LCN2), tissue inhibitor of metalloproteinase 1 (TIMP-1), interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS). In primary rat cardiomyocyte cultures Toll-like receptor 4 (TLR4) agonist- or PKCε/CN-dependent COX-2 induction occurred in coresident fibroblasts and was blocked by selective inhibition of CN or PKC α/ε or elimination of fibroblasts. Furthermore, ectopic expression of PKCε and CN in ARVFs showed that the effects on COX-2 expression are mediated by specific NFAT sites within the COX-2 promoter as confirmed by site-directed mutagenesis and chromatin immunoprecipitation (ChIP). Therefore, PKCε may negatively regulate adverse myocardial remodeling by cooperating with CN to downregulate fibrosis and induce transcription of cardioprotective wound healing genes, including COX-2
Host age and expression of genes involved in red blood cell invasion in Plasmodium falciparum field isolates
Plasmodium falciparum proteins involved in erythrocyte invasion
are main targets of acquired immunity and important vaccine
candidates. We hypothesized that anti-parasite immunity acquired
upon exposure would limit invasion-related gene (IRG) expression
and affect the clinical impact of the infection. 11 IRG
transcript levels were measured in P. falciparum isolates by
RT-PCR, and IgG/IgM against invasion ligands by Luminex(R), in
50 Mozambican adults, 25 children with severe malaria (SM) and
25 with uncomplicated malaria (UM). IRG expression differences
among groups and associations between IRG expression and
clinical/immunologic parameters were assessed. IRG expression
diversity was higher in parasites infecting children than adults
(p = 0.022). eba140 and ptramp expression decreased with age (p
= 0.003 and 0.007, respectively) whereas p41 expression
increased (p = 0.022). pfrh5 reduction in expression was abrupt
early in life. Parasite density decreased with increasing pfrh5
expression (p < 0.001) and, only in children, parasite
density increased with p41 expression (p = 0.007), and decreased
with eba175 (p = 0.013). Antibody responses and IRG expression
were not associated. In conclusion, IRG expression is associated
with age and parasite density, but not with specific antibody
responses in the acute phase of infection. Our results confirm
the importance of multi-antigen vaccines development to avoid
parasite immune escape when tested in malaria-exposed
individuals