Jorma Toppari

University of Copenhagen, København, Capital Region, Denmark

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Publications (316)1220.25 Total impact

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    ABSTRACT: Transcription factor GATA4 is expressed in somatic cells of the mammalian testis. Gene targeting studies in mice have shown that GATA4 is essential for proper differentiation and function of Sertoli cells. The role of GATA4 in Leydig cell development, however, remains controversial because targeted mutagenesis experiments in mice have not shown a consistent phenotype, possibly due to context-dependent effects or compensatory responses. We therefore undertook a reductionist approach to study the function of GATA4 in Leydig cells. Using microarray analysis and quantitative RT-PCR, we identified a set of genes that are downregulated or upregulated following siRNA-mediated silencing of Gata4 in the murine Leydig tumor cell line mLTC-1. These same genes were dysregulated when primary cultures of Gata4(flox/flox) adult Leydig cells were subjected to adenovirus-mediated cre-lox recombination in vitro. Among the downregulated genes were enzymes of the androgen biosynthetic pathway (Cyp11a1, Hsd3b1, Cyp17a1, Srd5a). Silencing of Gata4 expression in mLTC-1 cells was accompanied by reduced production of sex steroid precursors, as documented by mass spectrometric analysis. Comprehensive metabolomic analysis of GATA4-deficient mLTC-1 cells showed alteration of other metabolic pathways, notably glycolysis. GATA4-depleted mLTC-1 cells had reduced expression of glycolytic genes (Hk1, Gpi1, Pfkp, Pgam1), lower intracellular levels of ATP, and increased extracellular levels of glucose. Our findings suggest that GATA4 plays a pivotal role in Leydig cell function and provide novel insights into metabolic regulation in this cell type.
    Endocrinology 02/2015; · 4.72 Impact Factor
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    ABSTRACT: Several recent publications reflect debate on the issue of "endocrine disrupting chemicals" (EDCs), indicating that two seemingly mutually exclusive perspectives are being articulated separately and independently. Considering this, a group of scientists with expertise in basic science, medicine and risk assessment reviewed the various aspects of the debate to identify the most significant areas of dispute and to propose a path forward. We identified four areas of debate. The first is about the definitions for terms such as "endocrine disrupting chemical", "adverse effects", and "endocrine system". The second is focused on elements of hormone action including "potency", "endpoints", "timing", "dose" and "thresholds". The third addresses the information needed to establish sufficient evidence of harm. Finally, the fourth focuses on the need to develop and the characteristics of transparent, systematic methods to review the EDC literature. Herein we identify areas of general consensus and propose resolutions for these four areas that would allow the field to move beyond the current and, in our opinion, ineffective debate.
    Environmental health : a global access science source. 12/2014; 13(1):118.
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    ABSTRACT: Spermatozoa are produced during spermatogenesis as a result of mitotic proliferation, meiosis and cellular differentiation. Postmeiotic spermatids are exceptional cells given their haploid genome and remarkable sperm-specific structural transformations to compact and reshape the nucleus and to construct the flagellum and acrosome. These processes require delicate coordination and active communication between distinct cellular compartments. In this study, we elucidated the interplay between the haploid RNA regulation and the vesicular transport system. We identified a novel interaction between VPS26A/VPS35-containing retromer vesicles and the chromatoid body (CB), which is a large ribonucleoprotein (RNP) granule unique to haploid male germ cells. VPS26A/VPS35-positive vesicles were shown to be involved in the endosomal pathway, as well as in acrosomal formation that is dependent on the Golgi complex-derived vesicular trafficking. While the exact role of the retromer vesicles in the CB function remains unclear, our results suggest a direct functional link between vesicle transport and CB-mediated RNA regulation. Copyright © 2014. Published by Elsevier Ireland Ltd.
    Molecular and Cellular Endocrinology 12/2014; · 4.24 Impact Factor
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    ABSTRACT: Radiotherapy is a mainstay for treatment of many human cancer types, including head and neck squamous cell carcinoma (HNSCC). Thereby, it is clinically very relevant to understand the mechanisms determining radioresistance. Here, we identify CIP2A as an Oct4 target gene and provide evidence that they co-operate in radioresistance. Oct4 positively regulates CIP2A expression both in testicular cancer cell lines as well as in embryonic stem cells. To expand the relevance of these findings we show that Oct4 and CIP2A are co-expressed in CD24 positive side-population of patient-derived HNSCC cell lines. Most importantly, all Oct4 positive HNSCC patient samples were CIP2A positive and this double positivity was linked to poor differentiation level, and predicted for decreased patient survival among radiotherapy treated HNSCC patients. Oct4 and CIP2A expression was also linked with increased aggressiveness and radioresistancy in HNSCC cell lines. Together we demonstrate that CIP2A is a novel Oct4 target gene in stem cells and in human cancer cell lines. Clinically these results suggest that diagnostic evaluation of HNSCC tumors for Oct4 or Oct4/CIP2A positivity might help to predict HNSCC tumor radioresistancy. These results also identify both Oct4 and CIP2A as potential targets for radiosensitation.
    Oncotarget 12/2014; · 6.63 Impact Factor
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    ABSTRACT: PCB 180 is a persistent non-dioxin-like polychlorinated biphenyl (NDL-PCB) abundantly present in food and the environment. Risk characterization of NDL-PCBs is confounded by the presence of highly potent dioxin-like impurities. We used ultrapure PCB 180 to characterize its toxicity profile in a 28-day repeat dose toxicity study in young adult rats extended to cover endocrine and behavioral effects. Using a loading dose/maintenance dose regimen, groups of 5 males and 5 females were given total doses of 0, 3, 10, 30, 100, 300, 1000 or 1700 mg PCB 180/kg body weight by gavage. Dose-responses were analyzed using benchmark dose modeling based on dose and adipose tissue PCB concentrations. Body weight gain was retarded at 1700 mg/kg during loading dosing, but recovered thereafter. The most sensitive endpoint of toxicity that was used for risk characterization was altered open field behavior in females; i.e. increased activity and distance moved in the inner zone of an open field suggesting altered emotional responses to unfamiliar environment and impaired behavioral inhibition. Other dose-dependent changes included decreased serum thyroid hormones with associated histopathological changes, altered tissue retinoid levels, decreased hematocrit and hemoglobin, decreased follicle stimulating hormone and luteinizing hormone levels in males and increased expression of DNA damage markers in liver of females. Dose-dependent hypertrophy of zona fasciculata cells was observed in adrenals suggesting activation of cortex. There were gender differences in sensitivity and toxicity profiles were partly different in males and females. PCB 180 adipose tissue concentrations were clearly above the general human population levels, but close to the levels in highly exposed populations. The results demonstrate a distinct toxicological profile of PCB 180 with lack of dioxin-like properties required for assignment of WHO toxic equivalency factor. However, PCB 180 shares several toxicological targets with dioxin-like compounds emphasizing the potential for interactions.
    PLoS ONE 08/2014; 2934495. · 3.53 Impact Factor
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    ABSTRACT: PCB 180 is a persistent non-dioxin-like polychlorinated biphenyl (NDL-PCB) abundantly present in food and the environment. Risk characterization of NDL-PCBs is confounded by the presence of highly potent dioxin-like impurities. We used ultrapure PCB 180 to characterize its toxicity profile in a 28-day repeat dose toxicity study in young adult rats extended to cover endocrine and behavioral effects. Using a loading dose/maintenance dose regimen, groups of 5 males and 5 females were given total doses of 0, 3, 10, 30, 100, 300, 1000 or 1700 mg PCB 180/kg body weight by gavage. Dose-responses were analyzed using benchmark dose modeling based on dose and adipose tissue PCB concentrations. Body weight gain was retarded at 1700 mg/kg during loading dosing, but recovered thereafter. The most sensitive endpoint of toxicity that was used for risk characterization was altered open field behavior in females; i.e. increased activity and distance moved in the inner zone of an open field suggesting altered emotional responses to unfamiliar environment and impaired behavioral inhibition. Other dose-dependent changes included decreased serum thyroid hormones with associated histopathological changes, altered tissue retinoid levels, decreased hematocrit and hemoglobin, decreased follicle stimulating hormone and luteinizing hormone levels in males and increased expression of DNA damage markers in liver of females. Dose-dependent hypertrophy of zona fasciculata cells was observed in adrenals suggesting activation of cortex. There were gender differences in sensitivity and toxicity profiles were partly different in males and females. PCB 180 adipose tissue concentrations were clearly above the general human population levels, but close to the levels in highly exposed populations. The results demonstrate a distinct toxicological profile of PCB 180 with lack of dioxin-like properties required for assignment of WHO toxic equivalency factor. However, PCB 180 shares several toxicological targets with dioxin-like compounds emphasizing the potential for interactions.
    PLoS ONE 08/2014; 9(8):e104639. · 3.53 Impact Factor
  • Jorma Toppari
    Nature Reviews Urology 07/2014; · 4.79 Impact Factor
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    ABSTRACT: Although common reproductive problems, such as male infertility and testicular cancer, present in adult life, strong evidence exists that these reproductive disorders might have a fetal origin. The evidence is derived not only from large epidemiological studies that show birth-cohort effects with regard to testicular cancer, levels of testosterone and semen quality, but also from histopathological observations. Many infertile men have histological signs of testicular dysgenesis, including Sertoli-cell-only tubules, immature undifferentiated Sertoli cells, microliths and Leydig cell nodules. The most severe gonadal symptoms occur in patients with disorders of sexual development (DSDs) who have genetic mutations, in whom even sex reversal of individuals with a 46,XY DSD can occur. However, patients with severe DSDs might represent only a small proportion of DSD cases, with milder forms of testicular dysgenesis potentially induced by exposure to environmental and lifestyle factors. Interestingly, maternal smoking during pregnancy has a stronger effect on spermatogenesis than a man's own smoking. Other lifestyle factors such as alcohol consumption and obesity might also have a role. However, increasing indirect evidence exists that exposure to ubiquitous endocrine disrupting chemicals, present at measurable concentrations in individuals, might affect development of human fetal testis. If confirmed, health policies to prevent male reproductive problems should not only target adult men, but also pregnant women and their children.
    Nature Reviews Endocrinology 06/2014; · 11.03 Impact Factor
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    ABSTRACT: Background Humans are exposed to tributyltin (TBT), previously used as an antifouling paint in ships, mainly through fish consumption. As TBT is a known obesogen, we studied the association of placenta TBT and other organotin compounds (OTCs) with ponderal index (PI) and growth during the first 18 months of life in boys. Methods In a prospective Finnish study, 110 placenta samples were collected from mothers of boys born in 1997-1999 with (n = 55) and without (n = 55) cryptorchidism. To account for the original study design, linear regression, weighted for sampling fractions of boys with (121/55) and without (5677/55) cryptorchidism from the total cohort, was used to study the association between placenta OTCs and children's weight, length, growth rates and PI up to 18 months of age. Results Placenta TBT concentrations were above the limit of quantification (LOQ) in 99% of the samples. However, monobutyltin (MBT), dibutyltin (DBT) and triphenyltin (TPhT) concentrations were below LOQ in 90%, 35% and 57% of samples, respectively. Placenta TBT was positively associated (p = 0.024) with weight gain during the first three months of life, but no other significant associations were observed for weight or length gain. Also, no significant associations between placenta OTC concentrations and child length, weight or PI at any time point were found. Conclusions We observed a trend towards higher weight gain from birth to 3 months of age with increasing placenta TBT concentration. These results should be interpreted with caution because obesogenic effects in animal experiments were seen after in-utero TBT exposures to doses that were orders of magnitude higher. Also the number of study subjects included in this study was limited.
    Environmental Health 06/2014; 13(45). · 2.71 Impact Factor
    This article is viewable in ResearchGate's enriched format
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    ABSTRACT: The retinoblastoma protein (RB) is essential for normal cell cycle control. RB function depends, at least in part, on interactions with the E2F family of DNA-binding transcription factors (E2Fs). To study the role of RB in the adult testis, a Sertoli cell (SC)-specific Rb knockout mouse line (SC-RbKO) was generated using the Cre/loxP recombination system. SC-RbKO mice exhibited an age-dependent testicular atrophy, impaired fertility, severe SC dysfunction, and spermatogenic defects. Removal of Rb in SC induced aberrant SC cycling, dedifferentiation, and apoptosis. Here we show that E2F3 is the only E2F expressed in mouse SCs and that RB interacts with E2F3 during mouse testicular development. In the absence of RB, the other retinoblastoma family members p107 and p130 began interacting with E2F3 in the adult testes. In vivo silencing of E2F3 partially restored the SC maturation and survival as well as spermatogenesis in the SC-RbKO mice. These results point to RB as a key regulator of SC function in adult mice and that the RB/E2F3 pathway directs SC maturation, cell cycle quiescence, and RB protects SC from apoptosis.
    Cell Death & Disease 06/2014; 5:e1274. · 5.18 Impact Factor
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    ABSTRACT: Is there an association between alcohol intake and semen quality and serum reproductive hormones among healthy men from the USA and Europe?
    Human Reproduction 06/2014; · 4.59 Impact Factor
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    ABSTRACT: Testes descend to the scrotum normally before birth. When they fail to do so, the boy is cryptorchid and has an increased risk for testicular germ cell cancer and subfertility later in life. Early correction of maldescent by orchiopexy operation improves the spermatogenetic capacity of the testis but does not return the testicular cancer risk to the control level. Testicular descent is regulated by testis-derived hormones testosterone and insulin-like peptide 3. Cryptorchidism can therefore be considered a symptom of impaired testicular function that may also be linked to other testicular diseases, such as germ cell cancer and subfertility. Early orchiopexy can alleviate the effects of cryptorchidism on spermatogenesis, but alertness for testicular cancer should be maintained. In searching the genetic and environmental reasons for these diseases, it is useful to consider their connection with each other.
    Annales d Endocrinologie 04/2014; · 0.66 Impact Factor
  • Endocrine Abstracts. 04/2014;
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    ABSTRACT: Research on spermatogonia is hampered by complex architecture of the seminiferous tubule, poor viability of testicular tissue ex vivo and lack of physiologically relevant long-term culture systems. Therefore there is a need for an in vitro model that would enable long term survival and propagation of spermatogonia. We aimed at the most simplified approach to enable all different cell types within the seminiferous tubules to contribute to the creation of a niche for spermatogonia. In the present study we describe the establishment of a co-culture of mouse testicular cells that is based on proliferative and migratory activity of seminiferous tubule cells and does not involve separation, purification or differential plating of individual cell populations. The co-culture is composed of the constituents of testicular stem cell niche: Sertoli cells [identified by expression of Wilm's tumour antigen 1 (WT1) and secretion of glial cell line-derived neurotrophic factor, GDNF], peritubular myoid cells (expressing alpha smooth muscle actin, αSMA) and spermatogonia [expressing MAGE-B4, PLZF (promyelocytic leukaemia zinc finger), LIN28, Gpr125 (G protein-coupled receptor 125), CD9, c-Kit and Nanog], and can be maintained for at least five weeks. GDNF was found in the medium at a sufficient concentration to support proliferating spermatogonial stem cells (SSCs) that were able to start spermatogenic differentiation after transplantation to an experimentally sterile recipient testis. Gdnf mRNA levels were elevated by follicle-stimulating hormone (FSH) which shows that the Sertoli cells in the co-culture respond to physiological stimuli. After approximately 2-4 weeks of culture a spontaneous formation of cord-like structures was monitored. These structures can be more than 10 mm in length and branch. They are formed by peritubular myoid cells, Sertoli cells, fibroblasts and spermatogonia as assessed by gene expression profiling. In conclusion, we have managed to establish in vitro conditions that allow spontaneous reconstruction of testicular cellular microenvironments.
    PLoS ONE 03/2014; 9(3):e90088. · 3.53 Impact Factor
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    ABSTRACT: The genome of male germ cells is actively transcribed during spermatogenesis to produce phase-specific protein-coding mRNAs and a considerable amount of different noncoding RNAs. Ribonucleoprotein (RNP) granule-mediated RNA regulation provides a powerful means to secure the quality and correct expression of the requisite transcripts. Haploid spermatids are characterized by a unique, unusually large cytoplasmic granule, the chromatoid body (CB), which emerges during the switch between the meiotic and post-meiotic phases of spermatogenesis. To better understand the role of the CB in male germ cell differentiation, we isolated CBs from mouse testes and revealed its full RNA and protein composition. We showed that the CB is mainly composed of RNA-binding proteins and other proteins involved RNA regulation. The CB was loaded with RNA, including pachytene piRNAs, a diverse set of mRNAs, and a number of uncharacterized long noncoding transcripts. The CB was demonstrated to accumulate nascent RNA during all the steps of round spermatid differentiation. Our results revealed the CB as a large germ cell-specific RNP platform that is involved in the control of the highly complex transcriptome of haploid male germ cells.
    RNA 02/2014; · 4.62 Impact Factor
  • Helena E Virtanen, Jorma Toppari
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    ABSTRACT: Sexual differentiation starts with the development of bipotential gonads that further differentiate into testes or ovaries. The fetal testis secretes hormones that guide the differentiation of internal and external sex organs, whereas the fetal ovary remains rather inactive hormonally. Defects in gonadal differentiation or hormone secretion and action result in disorders of sex development (DSD). Testicular descent is a continuum that has often been described to occur in two main phases: the transabdominal phase and the inguinoscrotal phase. The first phase is according to animal studies dependent on Leydig cell-derived insulin-like peptide 3 (INSL3) that induces male-like development of the gubernaculum. This phase is rarely disrupted in man. The inguinoscrotal phase is dependent on androgens, also secreted by Leydig cells.
    Pediatric endocrinology reviews: PER 02/2014; 11 Suppl 2:206-13.
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    ABSTRACT: Les testicules descendent normalement avant la naissance dans le scrotum. Lorsque cette migration ne s’effectue pas, le garçon est cryptorchide et présente un risque accru de cancer testiculaire et d’hypofertilité ultérieure. Une correction précoce de cette anomalie par orchidopexie améliore la spermatogenèse mais n’annule pas le risque de cancer. La migration testiculaire est régulée par les hormones testiculaires, singulièrement la testosterone et le peptide insulin-like 3. Ainsi, la cryptorchidie peut être considérée comme un symptôme de l’altération de la fonction testiculaire qui peut être également liée à d’autres maladies testiculaires comme le cancer des cellules germinales et l’hypofertilité. Une orchiopexie précoce peut corriger les effets de la cryptorchidie sur la spermatogenèse, mais le dépistage du cancer testiculaire doit être maintenu. En recherchant les causes génétiques et environnementales de ces maladies, il est utile de prendre en compte leurs interrelations.
    Annales d'Endocrinologie. 01/2014;
  • Helena Virtanen, Jorma Toppari
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    ABSTRACT: Testes descend to the scrotum normally before birth. Testis-derived testosterone and insulin-like peptide 3 (INSL3) regulate the descent. Cryptorchid testis remains too high or in an abnormal location, which prevents its normal function. The incidence of cryptorchidism varies greatly: 1 to 8% in full-term newborn boys, and 1 to 2% at three months of age. Spontaneous descent can occur during the first three months. Defective androgen action can cause cryptorchidism, but usually the etiology remains unknown. Several factors (environment, genes and lifestyle) contribute to the risk of cryptorchidism. Acquired cryptorchidism also occurs during childhood. The recommended treatment of cryptorchidism is surgical orchidopexy.
    01/2014; 130(11):1086-92.
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    ABSTRACT: Geographical differences in occurrence of diseases in male reproductive organs including malformation in reproductive tract have been reported between Denmark and Finland. The reason for these differences is unknown, but differences in exposure to chemicals with endocrine disrupting abilities have been suggested. Among these chemicals are perfluoro¬alkylated substances (PFASs) a group of water and grease repellent chemicals used in outdoor clothes, cookware, food packaging and textiles. We therefore investigated differences in PFASs exposure levels between Denmark and Finland and associated PFASs levels in cord blood with congenital cryptorchidism. Boys from a joint ongoing prospective birth cohort study were included. We analyzed PFASs levels in cord blood serum samples from 29 Danish boys with congenital cryptorchidism; 30 healthy Danish matched controls recruited from 1997-2001, 30 Finnish cases and 78 Finnish healthy matched controls recruited from 1997-1999. Additionally, 48 Finnish cases recruited from 2000-2002 were included. PFOA and PFOS were detected in all 215 Danish and Finnish cord blood samples with significantly higher levels in Danish (medians; PFOA 2.6 ng/mL, PFOS 9.1 ng/mL) compared to Finnish (medians; PFOA: 2.1 ng/mL, PFOS 5.2 ng/mL) samples. We found no associations between cord blood PFOA and PFOS levels and congenital cryptorchidism after adjustment for confounders. Our data indicate that women in Denmark and Finland are generally exposed to PFOA and PFOS but with country differences in exposure levels. We found no statistical significant association between PFOA and PFOS levels in cord blood and congenital cryptorchidism, however, our study was small and larger studies are warranted.
    Reproduction 11/2013; · 3.26 Impact Factor
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Publication Stats

10k Citations
1,220.25 Total Impact Points

Institutions

  • 2008–2014
    • University of Copenhagen
      • • Faculty of Health and Medical Sciences
      • • Section of Biostatistics
      København, Capital Region, Denmark
    • University of Santiago de Compostela
      • Departamento de Fisiología
      Santiago de Compostela, Galicia, Spain
    • University of Tampere
      • Medical School
      Tampere, Western Finland, Finland
    • University of Kuopio
      Kuopio, Eastern Finland Province, Finland
    • University of California, Davis
      • Department of Environmental Toxicology
      Davis, CA, United States
    • Turku University Hospital
      • Department of Pediatrics
      Turku, Province of Western Finland, Finland
  • 1970–2014
    • University of Turku
      • • Department of Physiology
      • • Department of Paediatrics
      • • Institute of Biomedicine
      Turku, Province of Western Finland, Finland
  • 2013
    • National Institute for Health and Welfare, Finland
      • Department of Environmental Health
      Helsinki, Province of Southern Finland, Finland
    • St. Marianna University School of Medicine
      • Department of Urology
      Kawasaki, Kanagawa-ken, Japan
  • 2012
    • The University of Edinburgh
      • MRC Centre for Reproductive Health
      Edinburgh, Scotland, United Kingdom
  • 2009–2012
    • Technical University of Denmark
      • Center for Biological Sequence Analysis
      Copenhagen, Capital Region, Denmark
    • MTT Agrifood Research
      • Biotechnology and Food Research Unit
      Jokioinen, Province of Southern Finland, Finland
  • 2006–2012
    • IT University of Copenhagen
      København, Capital Region, Denmark
  • 1996–2012
    • University of Helsinki
      • • Institute of Biomedicine
      • • Hospital for Children and Adolescents and Biomedicum Helsinki
      Helsinki, Province of Southern Finland, Finland
  • 2011
    • Imperial College London
      • Department of Surgery and Cancer
      London, ENG, United Kingdom
  • 2007–2011
    • Region Hovedstaden
      Hillerød, Capital Region, Denmark
    • University of Washington Seattle
      • Department of Pediatrics
      Seattle, WA, United States
  • 2003–2006
    • University of Cordoba (Spain)
      • • Departamento de Biología Celular, Fisiología e Inmunología
      • • Facultad de Medicina
      Córdoba, Andalusia, Spain
  • 2005
    • Kaunas University of Technology
      Caunas, Kauno Apskritis, Lithuania
  • 2004
    • University of Southern Denmark
      Odense, South Denmark, Denmark
  • 2002–2004
    • Rigshospitalet
      • Department of Growth and Reproduction
      København, Capital Region, Denmark
  • 1989–2003
    • Harbor-UCLA Medical Center
      • Department of Pediatrics
      Torrance, California, United States
  • 2000
    • Odense University Hospital
      Odense, South Denmark, Denmark
  • 1991
    • Karolinska University Hospital
      • Endocrinology Unit
      Stockholm, Stockholm, Sweden
  • 1990
    • Stockholm University
      • Department of Biochemistry and Biophysics
      Stockholm, Stockholm, Sweden
  • 1989–1990
    • University of Southern California
      • Department of Obstetrics and Gynecology
      Los Angeles, CA, United States