A detailed protocol for a rapid analysis of testicular cell populations using flow cytometry

Accurate analysis and quantification of different testicular cell populations are of central importance in studies of male reproductive biology. The traditional histomorphometric and immunohistochemical methods remain the gold standard in studying the complex dynamics of the testicular tissue. Through past years advances have been made in the application of flow cytometry for the rapid analysis of testicular cell populations. Detection of DNA content and of surface antigens and fluorescent reporters have been widely used to analyze and sort cells. Detection of intracellular antigens can broaden the possibilities of applying flow cytometry in studies of male reproduction. Here, we report a detailed protocol for the preparation of rat testicular tissue for detection of intracellular antigens by flow cytometry, and a pipeline for subsequent data analysis and troubleshooting. Rat testicular ontogenesis was chosen as the experimental model to validate the performance of the assay using vimentin and gamma H2AX as intracellular markers for the somatic and spermatogenic cells, respectively. The results show that the assay is reproducible and recapitulates the rat testis ontogenesis.Peer reviewe

Decreased spermatogonial quantity in prepubertal boys with leukaemia treated with alkylating agents

Non peer reviewe

Serum 25-Hydroxyvitamin D Concentrations at Birth in Children Screened for HLA-DQB1 Conferred Risk for Type 1 Diabetes

Vitamin D has several effects on the immune system that might be of relevance for the pathogenesis of type 1 diabetes (T1D).To evaluate whether umbilical cord serum concentrations of 25-hydroxy-vitamin D (25[OH]D) differ in children developing either islet autoimmunity (IA) or overt T1D during childhood and adolescence.Umbilical cord serum samples from 764 children born from 1994 to 2004 with HLA-DQB1 conferred risk for T1D participating in the Type 1 Diabetes Prediction and Prevention Study were analyzed for 25(OH)D using an enzyme immunoassay.DIPP clinics in Turku, Oulu, and Tampere University Hospitals, Finland.Two hundred fifty children who developed T1D diabetes at a median age of 6.7 years (interquartile range [IQR] 4.0 to 10.1 years) and 132 additional case children who developed IA, i.e., positivity for multiple islet autoantibodies. Cases were matched for date of birth, gender, and area of birth with 382 control children who remained autoantibody negative. The median duration of follow up was 9.8 years (IQR 5.7 to 13.1 years).The median 25(OH)D concentrations.The median 25(OH)D concentration in cord serum was low [31.1 nmol/L (IQR 24.0 to 41.8); 88% <50 nmol/L], but not statistically different between children who developed T1D or IA and their control groups (P = 0.70). The levels were associated mainly with geographical location, year and month of birth, age of the mother, and maternal intake of vitamin D during pregnancy.The 25(OH)D concentrations at birth are not associated with the development of T1D during childhood.Peer reviewe

Serum 25-Hydroxyvitamin D Concentrations at Birth in Children Screened for HLA-DQB1 Conferred Risk for Type 1 Diabetes

Context: Vitamin D has several effects on the immune system that might be of relevance for the pathogenesis of type 1 diabetes (T1D).Objective: To evaluate whether umbilical cord serum concentrations of 25-hydroxy-vitamin D (25[OH]D) differ in children developing either islet autoimmunity (IA) or overt T1D during childhood and adolescence.Design: Umbilical cord serum samples from 764 children born from 1994 to 2004 with HLA-DQB1 conferred risk for T1 D participating in the Type 1 Diabetes Prediction and Prevention Study were analyzed for 25(OH)D using an enzyme immunoassay.Setting: DIPP clinics in Turku, Oulu, and Tampere University Hospitals, Finland.Participants: Two hundred fifty children who developed T1D diabetes at a median age of 6.7 years (interquartile range [IQR] 4.0 to 10.1 years) and 132 additional case children who developed IA, i.e., positivity for multiple islet autoantibodies. Cases were matched for date of birth, gender, and area of birth with 382 control children who remained autoantibody negative. The median duration of follow up was 9.8 years (IQR 5.7 to 13.1 years).Main Outcome Measure: The median 25(OH)D concentrations.Results: The median 25(OH)D concentration in cord serum was low [31.1 nmol/L (IQR 24.0 to 41.8); 88% Conclusions: The 25(OH)D concentrations at birth are not associated with the development of T1D during childhood.</div

Executing circular economy strategies in practice in Finland. Results and experiences from Circwaste project

A Europe-wide circular economy policy was launched in 2014 when the European Commission published the first strategic policy programme for circular economy. It was compiled to provide very comprehensive impacts and dimensions of sustainable development: sustainable growth and a climate neutral, resource efficient and competitive economy. The targets of a circular economy are that the value of products, materials and resources is maintained in the economy for as long as possible, economic growth is decoupled from resource use, generation of waste and environmental loads are minimised, and pressure on the Earth’s resources and biodiversity is minimised. The European Union is supporting the sustainability transition with research and development funding. In Finland, Circwaste – Towards Circular Economy is one of the biggest development projects accelerating the transition to a circular economy. During the period 2016–2020, the project has produced monitoring data on the development of circular economy and the sustainability of waste management, highlighted the circular economy concept, promoted stakeholder collaboration, supported strategic national processes, strengthened know-how and mainstreamed and concretised circular economy thinking. This interim report presents all the relevant results so far. It is crucial that data is produced from different angles on implementing the circular economy. More information is needed both to support decision making and on connections between and reflections on different factors. The key figures for Finland show quite clear coupling of the use of natural resources, waste amounts and economic growth. The circular material use rate is ca. 7%, which can be considered quite modest. Quantitative national targets for decreasing the use of natural resources are needed. Instead of country comparisons, the focus should be on trends in order to learn from the past and to identify the policy instruments needed to achieve the level aspired to. One of the key findings is the need for regional indicators and data for decisionmaking. The work done within Circwaste is the first effort towards a systematic monitoring scheme for monitoring circular economy regionally. The study showed that the production of regional waste data is challenging, that the estimated recycling rates have not increased adequately to reach the EU targets and that there could therefore be a need for municipallevel recycling targets. The transition to a circular economy also causes fundamental social changes in society. In the project, new indicators were developed for measuring social impacts: circular economy employment, education and employment for vulnerable groups, publicly shared resources, accessibility of recycling services and sustainable vehicle fuels. The first baseline data show advances towards the circular economy: the accessibility of waste management services has improved, the Finnish educational system has been able to respond quickly to the need for circular economy education, circular economy activities have potential for the employment of vulnerable groups and economic activities related to recycling, repair and reuse have grown. The regions and municipalities emerge as key actors in facilitating a socially just transition towards a circular economy. The study on innovative material processing technologies gathered data on technologies for elemental recycling, especially for plastic waste but also for making new fibres from textiles waste. Financial issues are key to the survival of these technologies and there is a need for governmental financial support. Public procurers can be considered key players in the circular economy, creating demand for more sustainable products and services. Implementing circular economy in municipalities requires commitment, financial planning, interaction with regional actors and inclusion of circular economy in financial rules. The construction sector is a major consumer of natural resources, but the municipalities can make construction more sustainable through public procurements and planning. As buyers, they can require the use of recycled raw materials and soils in construction projects. Obligations for ecological compensation and goals of no net loss of biodiversity would decrease the pressure on natural resources. To support municipalities in their work, a national organisation for providing municipal auditing, development, education and business support services could be established. Employing circular economy experts in each municipality to work as crossadministrative coordinators could enhance the transition. The project has created a lot of political, theoretical and practical content on the concept and field of circular economy. The next steps are to further develop and widen, as well as deepen, the results and to provide national support in searching for answers and solutions for decreasing the use of natural resources, achieving the MSW recycling targets and creating a more sustainable society

Age- and cell-related gene expression of aromatase and estrogen receptors in the rat testis

International audienceSpermatogenesis is a complex and coordinated process leading to the formation of spermatozoa. This event, which is under the control of numerous endocrine and paracrine factors, seems to also be controlled by estrogens which exert their effects via nuclear estrogen receptors (ESRs) ESR1 and ESR2. Estrogens are synthesized by aromatase which is biologically expressed in the rat testis. The objective of our study was to clarify the gene expression patterns of aromatase and ESRs according to age and in the two compartments of the adult rat testis. In the adult, transcripts of aromatase vary according to the germ cell type and to the stages of seminiferous epithelium, a maximum being observed at stage I. The ESR1 gene is highly expressed in the adult testis and in stages from VIIc-d to XIV. Moreover, both ESR mRNA levels are higher in purified round spermatids than in pachytene spermatocytes, suggesting a putative role of estrogens in the haploid steps of spermatogenesis. The variability of the results in the expression of both ESRs led us to explore the putative presence of variants in the rat testis. Concerning ESR1, we have shown the presence of the full-length form and of one isoform with exon 4 deleted. For ESR2, besides the wild type, three isoforms were observed: one with exon 3 deleted, another with an insertion of 54 nucleotides, and the last one with both modifications. Therefore, the stage-regulated expression of aromatase and ESR1 genes in the rat testis suggests a likely role of estrogens in spermatogenesis