Desenvolupament de mètodes experimentals de cultius d’amastigots de Leishmania spp. per millorar la quantificació dels resultats experimentals

Leishmaniases are a group of diseases that cause thousands of deaths every year in the Southcountries. These diseases are caused by a protozoa of the genus Leishmania spp., which develops its life cycle between a vertebrate host and an the invertebrate vector through which they are transmitted (sand flies). Leishmania spp. belongs to the Trypanosomatidae family, which also includes species of the genus Trypanosoma. It causes the different forms of tripanosomiasis, known as Chagas disease in America and sleeping illness in Africa. Both of them are considered by the World Health Organization (WHO) as one of the 17 Neglected Tropical Diseases. The purpose of this work is to go further and deeper in the quantification methods of the experimental results from in vitro cultures of Leishmania spp. amastigotes (i.e., the intracellular form of the vertebrate stage). The experimental work has been done in the Parasitology Section of the Department of Health Microbiology and Parasitology of the Faculty of Pharmacy (University of Barcelona). Several assays were carried out, in which macrophages collected from murine animals and from the cell line RAW264.7 were infected with promastigotes (extracellular form of the vector stage) of different species of Leishmania (L. infantum, L. major and L. braziliensis). Different observations on the procedures of collecting murine cells, maintaining and monitoring the cultures were pointed. A common time pattern was established for both types of macrophages. The identification of intermediate forms (promastigotes changing into amastigotes) and the time when amastigotes number reached its maximum peak (72 h) were set up. The digitalization of the slides from the cultures was done at Barcelona School of Agricultural Engineering (ESAB). More than 700 images were collected. In order to allow for a better understanding the images were captured with 20 and 50 magnification objectives lens. The use of several experimental procedures allowed to analyse the differences between them and to improve the methodology. An experimental protocol for the cultures and their digitalization was proposed to have a better processing of the obtained information. The need for improving the experimental protocols and developing an image analysis technique to process the information obtained from them were shown, in order to have a better understanding of the parasite. At the same time, it is necessary to work on an Individual based Model (IbM) in order to introduce the system complexity at an individual level.La leishmaniosi engloba un conjunt de malalties que causen milers de morts anualment als països del Sud. Aquesta està causada per protozous del gènere Leishmania spp., que desenvolupen el seu cicle de vida entre un hoste vertebrat i un vector invertebrat mitjançant el qual es transmeten (flebòtoms, família Phlebotominae). Leishmania spp. pertany a la família Trypanosomatidae, en la que també s'inclouen les espècies del gènere Trypanosoma responsables de les diferents formes de tripanosomiasis, coneguda com la malaltia de Chagas al continent Americà i com la malaltia de la son al continent Africà. Totes elles formen part de les 17 malalties tropicals que la Organització Mundial de la Salut (OMS) identifica com a desateses. L'objectiu principal d'aquest Treball Final de Grau és avançar en les metodologies de quantificació dels resultats experimentals procedents dels cultius in vitro d'amastigots de Leishmania spp. (forma intracel·lular, pròpia de l'hoste vertebrat). El treball experimental s'ha realitzat sota la direcció de la Secció de Parasitologia del Departament de Biologia, Sanitat i Medi Ambient a la Facultat de Farmàcia i Ciències de l'Alimentació (Universitat de Barcelona). S'han dut a terme diferents assajos en els que s'han infectat macròfags d'origen peritoneal murí i de línia cel·lular RAW 264.7 amb promastigots (forma extracel·lular, pròpia de l'hoste invertebrat) de diferents espècies del gènere Leishmania (L. infantum, L. major i L. braziliensis). Al llarg dels assajos s'han realitzat observacions sobre els procediments utilitzats per l'obtenció de les cèl·lules d'origen peritoneal murí, la infecció i el manteniment dels cultius. A partir del seu seguiment s'ha pogut establir un patró temporal comú independentment del tipus de macròfags infectats, en el que s'han identificat formes intermèdies extracel·lulars(promastigots que varien la seva morfologia cap a amastigots) i s'ha pogut determinar que el nombre d'amastigots és màxim a les 72 hores. La digitalització de les preparacions procedents dels cultius s'ha realitzat a l'Escola Superior d'Agricultura de Barcelona (ESAB). S'ha elaborat una col·lecció de més de 700 imatges. Aquestes s'han capturat a 20 i 50 augments per facilitar la comprensió del sistema a nivell dels macròfags i del cultiu. L'ús de diferents procediments experimentals ha permès avaluar els seus efectes i millorar la metodologia utilitzada. En aquesta línia, hem definit un protocol experimental, tan dels cultius com de la seva digitalització, amb l'objectiu de facilitar el posterior processat de la informació obtinguda. Per seguir avançant en la comprensió del paràsit, cal seguir treballant per millorar els procediments experimentals i desenvolupar una tècnica d'anàlisi d'imatges que ens permeti processar la informació que se n'obté. De manera paral·lela, cal elaborar un model basat en l'individu (Individual based Model; IbM) que ens permeti introduir la complexitat del sistema de manera gradual a nivell individual.La leishmaniosis engloba un conjunto de enfermedades que causan miles de muertes anualmente en los países del Sud. Está causada por protozoos del género Leishmania spp., que desarrollan su ciclo de vida entre un hospedador vertebrado y un vector invertebrado mediante el que se transmiten. Leishmania spp. pertenece a la familia Trypanosomatidae, en la que también se incluyen las especies del género Trypanosoma responsables de las diferentes formas de tripanosomiasis, conocida como la enfermedad de Chagas en el continente Americano y cómo la enfermedad del sueño en el continente Africano. Todas ellas forman parte de las 17 enfermedades tropicales que la Organización Mundial de la Salud (OMS) identifica como desatendidas. El objetivo principal de este Trabajo de Fin de Grado es avanzar en las metodologías de cuantificación de los resultados experimentales procedentes de los cultivos in vitro de amastigotes de Leishmania spp. (forma intracelular, propia del hospedador vertebrado). El trabajo experimental se ha realizado bajo la dirección de la Sección de Parasitologia del Departamento de Biología, Sanidad y Medio Ambiente de la Facultad de Farmacia y Ciencias de la Alimentación (Universidad de Barcelona). Se han llevado a cabo diferentes ensayos en los que se han infectado macrófagos de origen peritoneal murino y de línea celular RAW 264.7 con promastigotes (forma extracelular, propia del hospedador invertebrado) de diferentes especies de Leishmania spp. (L. infantum, L. major y L.braziliensis). En el transcurso de los ensayos se han realizado observaciones sobre los procedimientos utilizados para la obtención de células de origen peritoneal murino, la infección y el mantenimiento de los cultivos. A partir de su observación se ha podido establecer un patrón temporal común independientemente del tipo de macrófago utilizado, en el que se han identificado formas intermedias extracelulares (promastigotes que varían su morfología hacia amastigotes) y se ha podido determinar que el número de amastigotes es máximo a las 72 horas. La digitalización de las preparaciones de los cultivos se ha realizado en la Escuela Superior de Agricultura de Barcelona (ESAB). Se ha elaborado una colección de más de 700 imágenes. Éstas se han capturado a 20 y 50 aumentos para facilitar la comprensión del sistema a nivel de los macrófagos y el cultivo. El uso de diferentes procedimientos experimentales ha permitido avaluar sus efectos y mejorar la metodología utilizada. En esta línea, hemos definido un protocolo experimental, no sólo de los cultivos sino que también de su digitalización, con el objetivo de facilitar el posterior procesado de la información obtenida. Para seguir avanzando en la comprensión del parásito, es necesario seguir trabajando para mejorar los procedimientos experimentales y desarrollar una técnica de análisis de imágenes que nos permita procesar la información que se obtiene de ellos. Paralelamente, es necesario elaborar un modelo basado en el individuo (Individual based Model; IbM) que nos permita introducir de manera gradual la complejidad del sistema a nivel individual

Trends of use and characterisation of anti-dementia drugs users:a large multinational-network population-based study

Background: An updated time-trend analysis of anti-dementia drugs (ADDs) is lacking. The aim of this study is to assess the incident rate (IR) of ADD in individuals with dementia using real-world data. Setting: Primary care data (country/database) from the UK/CPRD-GOLD (2007–20), Spain/SIDIAP (2010–20) and the Netherlands/IPCI (2008–20), standardised to a common data model. Methods: Cohort study. Participants: dementia patients ≥40 years old with ≥1 year of previous data. Follow-up: until the end of the study period, transfer out of the catchment area, death or incident prescription of rivastigmine, galantamine, donepezil or memantine. Other variables: age/sex, type of dementia, comorbidities. Statistics: overall and yearly age/sex IR, with 95% confidence interval, per 100,000 person-years (IR per 105 PY (95%CI)). Results: We identified a total of (incident anti-dementia users/dementia patients) 41,024/110,642 in UK/CPRD-GOLD, 51,667/134,927 in Spain/SIDIAP and 2,088/17,559 in the Netherlands/IPCI. In the UK, IR (per 105 PY (95%CI)) of ADD decreased from 2007 (30,829 (28,891–32,862)) to 2010 (17,793 (17,083–18,524)), then increased up to 2019 (31,601 (30,483 to 32,749)) and decrease in 2020 (24,067 (23,021–25,148)). In Spain, IR (per 105 PY (95%CI)) of ADD decreased by 72% from 2010 (51,003 (49,199–52,855)) to 2020 (14,571 (14,109–15,043)). In the Netherlands, IR (per 105 PY (95%CI)) of ADD decreased by 77% from 2009 (21,151 (14,967–29,031)) to 2020 (4763 (4176–5409)). Subjects aged ≥65–79 years and men (in the UK and the Netherlands) initiated more frequently an ADD. Conclusions: Treatment of dementia remains highly heterogeneous. Further consensus in the pharmacological management of patients living with dementia is urgently needed.</p

Trends of use and characterisation of anti-dementia drugs users:a large multinational-network population-based study

Background: An updated time-trend analysis of anti-dementia drugs (ADDs) is lacking. The aim of this study is to assess the incident rate (IR) of ADD in individuals with dementia using real-world data. Setting: Primary care data (country/database) from the UK/CPRD-GOLD (2007–20), Spain/SIDIAP (2010–20) and the Netherlands/IPCI (2008–20), standardised to a common data model. Methods: Cohort study. Participants: dementia patients ≥40 years old with ≥1 year of previous data. Follow-up: until the end of the study period, transfer out of the catchment area, death or incident prescription of rivastigmine, galantamine, donepezil or memantine. Other variables: age/sex, type of dementia, comorbidities. Statistics: overall and yearly age/sex IR, with 95% confidence interval, per 100,000 person-years (IR per 105 PY (95%CI)). Results: We identified a total of (incident anti-dementia users/dementia patients) 41,024/110,642 in UK/CPRD-GOLD, 51,667/134,927 in Spain/SIDIAP and 2,088/17,559 in the Netherlands/IPCI. In the UK, IR (per 105 PY (95%CI)) of ADD decreased from 2007 (30,829 (28,891–32,862)) to 2010 (17,793 (17,083–18,524)), then increased up to 2019 (31,601 (30,483 to 32,749)) and decrease in 2020 (24,067 (23,021–25,148)). In Spain, IR (per 105 PY (95%CI)) of ADD decreased by 72% from 2010 (51,003 (49,199–52,855)) to 2020 (14,571 (14,109–15,043)). In the Netherlands, IR (per 105 PY (95%CI)) of ADD decreased by 77% from 2009 (21,151 (14,967–29,031)) to 2020 (4763 (4176–5409)). Subjects aged ≥65–79 years and men (in the UK and the Netherlands) initiated more frequently an ADD. Conclusions: Treatment of dementia remains highly heterogeneous. Further consensus in the pharmacological management of patients living with dementia is urgently needed.</p

Effectiveness of COVID-19 vaccines against severe COVID-19 among patients with cancer in Catalonia, Spain

Patients with cancer were excluded from pivotal randomized clinical trials of COVID-19 vaccine products, and available observational evidence on vaccine effectiveness (VE) focused mostly on mild, and not severe COVID-19, which is the ultimate goal of vaccination for high-risk groups. Here, using primary care electronic health records from Catalonia, Spain (SIDIAP), we built two large cohorts of vaccinated and matched control cancer patients with a primary vaccination scheme (n = 184,744) and a booster (n = 108,534). Most patients received a mRNA-based product in primary (76.2%) and booster vaccination (99.9%). Patients had 51.8% (95% CI 40.3%−61.1%) and 58.4% (95% CI 29.3%−75.5%) protection against COVID-19 hospitalization and COVID-19 death respectively after full vaccination (two-doses) and 77.9% (95% CI 69.2%−84.2%) and 80.2% (95% CI 63.0%−89.4%) after booster. Compared to primary vaccination, the booster dose provided higher peak protection during follow-up. Calibration of VE estimates with negative outcomes, and sensitivity analyses with slight different population and COVID-19 outcomes definitions provided similar results. Our results confirm the role of primary and booster COVID-19 vaccination in preventing COVID-19 severe events in patients with cancer and highlight the need for the additional dose in this population.</p

Impact of the COVID-19 pandemic on diagnoses of common mental health disorders in adults in Catalonia, Spain : a population-based cohort study

To investigate how trends in incidence of anxiety and depressive disorders have been affected by the COVID-19 pandemic. Population-based cohort study. Retrospective cohort study from 2018 to 2021 using the Information System for Research in Primary Care (SIDIAP) database in Catalonia, Spain. 3 640 204 individuals aged 18 or older in SIDIAP on 1 March 2018 with no history of anxiety and depressive disorders. The incidence of anxiety and depressive disorders during the prelockdown period (March 2018-February 2020), lockdown period (March-June 2020) and postlockdown period (July 2020-March 2021) was calculated. Forecasted rates over the COVID-19 periods were estimated using negative binomial regression models based on prelockdown data. The percentage of reduction was estimated by comparing forecasted versus observed events, overall and by sex, age and socioeconomic status. The incidence rates per 100 000 person-months of anxiety and depressive disorders were 151.1 (95% CI 150.3 to 152.0) and 32.3 (31.9 to 32.6), respectively, during the prelockdown period. We observed an increase of 37.1% (95% prediction interval 25.5 to 50.2) in incident anxiety diagnoses compared with the expected in March 2020, followed by a reduction of 15.8% (7.3 to 23.5) during the postlockdown period. A reduction in incident depressive disorders occurred during the lockdown and postlockdown periods (45.6% (39.2 to 51.0) and 22.0% (12.6 to 30.1), respectively). Reductions were higher among women during the lockdown period, adults aged 18-34 years and individuals living in the most deprived areas. The COVID-19 pandemic in Catalonia was associated with an initial increase in anxiety disorders diagnosed in primary care but a reduction in cases as the pandemic continued. Diagnoses of depressive disorders were lower than expected throughout the pandemic

The impact of the UK COVID-19 lockdown on the screening, diagnostics and incidence of breast, colorectal, lung and prostate cancer in the UK: a population-based cohort study

Introduction: The COVID-19 pandemic had collateral effects on many health systems. Cancer screening and diagnostic tests were postponed, resulting in delays in diagnosis and treatment. This study assessed the impact of the pandemic on screening, diagnostics and incidence of breast, colorectal, lung, and prostate cancer; and whether rates returned to pre-pandemic levels by December, 2021. Methods: This is a cohort study of electronic health records from the United Kingdom (UK) primary care Clinical Practice Research Datalink (CPRD) GOLD database. The study included individuals registered with CPRD GOLD between January, 2017 and December, 2021, with at least 365 days of clinical history. The study focused on screening, diagnostic tests, referrals and diagnoses of first-ever breast, colorectal, lung, and prostate cancer. Incidence rates (IR) were stratified by age, sex, and region, and incidence rate ratios (IRR) were calculated to compare rates during and after lockdown with rates before lockdown. Forecasted rates were estimated using negative binomial regression models. Results: Among 5,191,650 eligible participants, the first lockdown resulted in reduced screening and diagnostic tests for all cancers, which remained dramatically reduced across the whole observation period for almost all tests investigated. There were significant IRR reductions in breast (0.69 [95% CI: 0.63-0.74]), colorectal (0.74 [95% CI: 0.67-0.81]), and prostate (0.71 [95% CI: 0.66-0.78]) cancer diagnoses. IRR reductions for lung cancer were non-significant (0.92 [95% CI: 0.84-1.01]). Extrapolating to the entire UK population, an estimated 18,000 breast, 13,000 colorectal, 10,000 lung, and 21,000 prostate cancer diagnoses were missed from March, 2020 to December, 2021. Discussion: The UK COVID-19 lockdown had a substantial impact on cancer screening, diagnostic tests, referrals, and diagnoses. Incidence rates remained significantly lower than pre-pandemic levels for breast and prostate cancers and associated tests by December, 2021. Delays in diagnosis are likely to have adverse consequences on cancer stage, treatment initiation, mortality rates, and years of life lost. Urgent strategies are needed to identify undiagnosed cases and address the long-term implications of delayed diagnoses

IncidencePrevalence:An R package to calculate population-level incidence rates and prevalence using the OMOP common data model

Purpose: Real-world data (RWD) offers a valuable resource for generating population-level disease epidemiology metrics. We aimed to develop a well-tested and user-friendly R package to compute incidence rates and prevalence in data mapped to the observational medical outcomes partnership (OMOP) common data model (CDM). Materials and Methods: We created IncidencePrevalence, an R package to support the analysis of population-level incidence rates and point- and period-prevalence in OMOP-formatted data. On top of unit testing, we assessed the face validity of the package. To do so, we calculated incidence rates of COVID-19 using RWD from Spain (SIDIAP) and the United Kingdom (CPRD Aurum), and replicated two previously published studies using data from the Netherlands (IPCI) and the United Kingdom (CPRD Gold). We compared the obtained results to those previously published, and measured execution times by running a benchmark analysis across databases. Results: IncidencePrevalence achieved high agreement to previously published data in CPRD Gold and IPCI, and showed good performance across databases. For COVID-19, incidence calculated by the package was similar to public data after the first-wave of the pandemic. Conclusion: For data mapped to the OMOP CDM, the IncidencePrevalence R package can support descriptive epidemiological research. It enables reliable estimation of incidence and prevalence from large real-world data sets. It represents a simple, but extendable, analytical framework to generate estimates in a reproducible and timely manner.</p

Famílies botàniques de plantes medicinals

Facultat de Farmàcia, Universitat de Barcelona. Ensenyament: Grau de Farmàcia, Assignatura: Botànica Farmacèutica, Curs: 2013-2014, Coordinadors: Joan Simon, Cèsar Blanché i Maria Bosch.Els materials que aquí es presenten són els recull de 175 treballs d’una família botànica d’interès medicinal realitzats de manera individual. Els treballs han estat realitzat per la totalitat dels estudiants dels grups M-2 i M-3 de l’assignatura Botànica Farmacèutica durant els mesos d’abril i maig del curs 2013-14. Tots els treballs s’han dut a terme a través de la plataforma de GoogleDocs i han estat tutoritzats pel professor de l’assignatura i revisats i finalment co-avaluats entre els propis estudiants. L’objectiu principal de l’activitat ha estat fomentar l’aprenentatge autònom i col·laboratiu en Botànica farmacèutica

Desenvolupament de mètodes experimentals de cultius d’amastigots de Leishmania spp. per millorar la quantificació dels resultats experimentals

Leishmaniases are a group of diseases that cause thousands of deaths every year in the Southcountries. These diseases are caused by a protozoa of the genus Leishmania spp., which develops its life cycle between a vertebrate host and an the invertebrate vector through which they are transmitted (sand flies). Leishmania spp. belongs to the Trypanosomatidae family, which also includes species of the genus Trypanosoma. It causes the different forms of tripanosomiasis, known as Chagas disease in America and sleeping illness in Africa. Both of them are considered by the World Health Organization (WHO) as one of the 17 Neglected Tropical Diseases. The purpose of this work is to go further and deeper in the quantification methods of the experimental results from in vitro cultures of Leishmania spp. amastigotes (i.e., the intracellular form of the vertebrate stage). The experimental work has been done in the Parasitology Section of the Department of Health Microbiology and Parasitology of the Faculty of Pharmacy (University of Barcelona). Several assays were carried out, in which macrophages collected from murine animals and from the cell line RAW264.7 were infected with promastigotes (extracellular form of the vector stage) of different species of Leishmania (L. infantum, L. major and L. braziliensis). Different observations on the procedures of collecting murine cells, maintaining and monitoring the cultures were pointed. A common time pattern was established for both types of macrophages. The identification of intermediate forms (promastigotes changing into amastigotes) and the time when amastigotes number reached its maximum peak (72 h) were set up. The digitalization of the slides from the cultures was done at Barcelona School of Agricultural Engineering (ESAB). More than 700 images were collected. In order to allow for a better understanding the images were captured with 20 and 50 magnification objectives lens. The use of several experimental procedures allowed to analyse the differences between them and to improve the methodology. An experimental protocol for the cultures and their digitalization was proposed to have a better processing of the obtained information. The need for improving the experimental protocols and developing an image analysis technique to process the information obtained from them were shown, in order to have a better understanding of the parasite. At the same time, it is necessary to work on an Individual based Model (IbM) in order to introduce the system complexity at an individual level.La leishmaniosi engloba un conjunt de malalties que causen milers de morts anualment als països del Sud. Aquesta està causada per protozous del gènere Leishmania spp., que desenvolupen el seu cicle de vida entre un hoste vertebrat i un vector invertebrat mitjançant el qual es transmeten (flebòtoms, família Phlebotominae). Leishmania spp. pertany a la família Trypanosomatidae, en la que també s'inclouen les espècies del gènere Trypanosoma responsables de les diferents formes de tripanosomiasis, coneguda com la malaltia de Chagas al continent Americà i com la malaltia de la son al continent Africà. Totes elles formen part de les 17 malalties tropicals que la Organització Mundial de la Salut (OMS) identifica com a desateses. L'objectiu principal d'aquest Treball Final de Grau és avançar en les metodologies de quantificació dels resultats experimentals procedents dels cultius in vitro d'amastigots de Leishmania spp. (forma intracel·lular, pròpia de l'hoste vertebrat). El treball experimental s'ha realitzat sota la direcció de la Secció de Parasitologia del Departament de Biologia, Sanitat i Medi Ambient a la Facultat de Farmàcia i Ciències de l'Alimentació (Universitat de Barcelona). S'han dut a terme diferents assajos en els que s'han infectat macròfags d'origen peritoneal murí i de línia cel·lular RAW 264.7 amb promastigots (forma extracel·lular, pròpia de l'hoste invertebrat) de diferents espècies del gènere Leishmania (L. infantum, L. major i L. braziliensis). Al llarg dels assajos s'han realitzat observacions sobre els procediments utilitzats per l'obtenció de les cèl·lules d'origen peritoneal murí, la infecció i el manteniment dels cultius. A partir del seu seguiment s'ha pogut establir un patró temporal comú independentment del tipus de macròfags infectats, en el que s'han identificat formes intermèdies extracel·lulars(promastigots que varien la seva morfologia cap a amastigots) i s'ha pogut determinar que el nombre d'amastigots és màxim a les 72 hores. La digitalització de les preparacions procedents dels cultius s'ha realitzat a l'Escola Superior d'Agricultura de Barcelona (ESAB). S'ha elaborat una col·lecció de més de 700 imatges. Aquestes s'han capturat a 20 i 50 augments per facilitar la comprensió del sistema a nivell dels macròfags i del cultiu. L'ús de diferents procediments experimentals ha permès avaluar els seus efectes i millorar la metodologia utilitzada. En aquesta línia, hem definit un protocol experimental, tan dels cultius com de la seva digitalització, amb l'objectiu de facilitar el posterior processat de la informació obtinguda. Per seguir avançant en la comprensió del paràsit, cal seguir treballant per millorar els procediments experimentals i desenvolupar una tècnica d'anàlisi d'imatges que ens permeti processar la informació que se n'obté. De manera paral·lela, cal elaborar un model basat en l'individu (Individual based Model; IbM) que ens permeti introduir la complexitat del sistema de manera gradual a nivell individual.La leishmaniosis engloba un conjunto de enfermedades que causan miles de muertes anualmente en los países del Sud. Está causada por protozoos del género Leishmania spp., que desarrollan su ciclo de vida entre un hospedador vertebrado y un vector invertebrado mediante el que se transmiten. Leishmania spp. pertenece a la familia Trypanosomatidae, en la que también se incluyen las especies del género Trypanosoma responsables de las diferentes formas de tripanosomiasis, conocida como la enfermedad de Chagas en el continente Americano y cómo la enfermedad del sueño en el continente Africano. Todas ellas forman parte de las 17 enfermedades tropicales que la Organización Mundial de la Salud (OMS) identifica como desatendidas. El objetivo principal de este Trabajo de Fin de Grado es avanzar en las metodologías de cuantificación de los resultados experimentales procedentes de los cultivos in vitro de amastigotes de Leishmania spp. (forma intracelular, propia del hospedador vertebrado). El trabajo experimental se ha realizado bajo la dirección de la Sección de Parasitologia del Departamento de Biología, Sanidad y Medio Ambiente de la Facultad de Farmacia y Ciencias de la Alimentación (Universidad de Barcelona). Se han llevado a cabo diferentes ensayos en los que se han infectado macrófagos de origen peritoneal murino y de línea celular RAW 264.7 con promastigotes (forma extracelular, propia del hospedador invertebrado) de diferentes especies de Leishmania spp. (L. infantum, L. major y L.braziliensis). En el transcurso de los ensayos se han realizado observaciones sobre los procedimientos utilizados para la obtención de células de origen peritoneal murino, la infección y el mantenimiento de los cultivos. A partir de su observación se ha podido establecer un patrón temporal común independientemente del tipo de macrófago utilizado, en el que se han identificado formas intermedias extracelulares (promastigotes que varían su morfología hacia amastigotes) y se ha podido determinar que el número de amastigotes es máximo a las 72 horas. La digitalización de las preparaciones de los cultivos se ha realizado en la Escuela Superior de Agricultura de Barcelona (ESAB). Se ha elaborado una colección de más de 700 imágenes. Éstas se han capturado a 20 y 50 aumentos para facilitar la comprensión del sistema a nivel de los macrófagos y el cultivo. El uso de diferentes procedimientos experimentales ha permitido avaluar sus efectos y mejorar la metodología utilizada. En esta línea, hemos definido un protocolo experimental, no sólo de los cultivos sino que también de su digitalización, con el objetivo de facilitar el posterior procesado de la información obtenida. Para seguir avanzando en la comprensión del parásito, es necesario seguir trabajando para mejorar los procedimientos experimentales y desarrollar una técnica de análisis de imágenes que nos permita procesar la información que se obtiene de ellos. Paralelamente, es necesario elaborar un modelo basado en el individuo (Individual based Model; IbM) que nos permita introducir de manera gradual la complejidad del sistema a nivel individual