P-440 Impact of electrospun scaffold topology on the performance of in-vitro Folliculogenesis applied to preantral ovine follicles

Study question How to improve in-vitro Folliculogenesis (ivF) protocols to address the enlarged demand of fertility preservation? Summary answer Tissue engineering-based approach opens new frontiers for ivF improving 3D-technologies addressed to support immature-ovarian-follicle-growth to obtain an increased number of competent oocytes enrolled in Assisted-Reproductive-Technology. What is known already ivF is a promising Assisted-Reproductive-Technology (ART) for preserving and restoring fertility. This technology potentially reproduces the early stages of folliculogenesis and oogenesis in-vitro allowing to move a large amount of oocyte on individual basis towards the validated protocol of in-vitro maturation/in-vitro fertilization (IVM/IVF). The current availability of biocompatible-supporting materials offers the challenging opportunity to mimic the native organ stroma in order to better reproduce the 3D environmental conditions leading to synergic follicles-oocyte development in-vitro with the aim to improve the performance of ivF in translational large sized mammal models. Study design, size, duration The present research aimed to compare preantral (PA) follicles culture on two different typologies of scaffolds fabricated using PCL(poly(epsilon caprolactone)), respectively made with patterned and randomly aligned fibers (PCL-Patterned/PCL-Randomic) with a standardized-single-follicle scaffold-free-method (3D-oil), widely validated on ovine model (Cecconi et al., 2004). The culture outcomes are compared analyzing follicle/oocyte growth, percentage of antrum differentiation and the incidence of meiotic competence, by exposing ivF growing oocytes to IVM protocol. Participants/materials, setting, methods PA follicles (mean size diameter: 250±4μm), mechanically isolated from slaughterhoused lamb ovaries, were individually cultured on electrospun PCL scaffolds (patterned vs randomic) or using the 3D-oil method. ivF were cultured alphaMEM-Fetal Bovine Serum free medium (5% Knockout Serum Replacement) supplemented with 4 IU/mL of equine Chorionic Gonadotropin (Di Berardino et al., 2021). At the end of ivF (14-days) the fully-grown oocytes isolated from early-antral follicles were tested on IVM. Main results and the role of chance PCL-Patterned electrospun scaffolds were able to strongly support a synergic oocyte and follicular growth. The 3D culture on Patterned electrospun scaffold supported the highest viability of follicles (87.5% vs 63% under 3D-oil conditions). On the contrary, the highest incidence of degenerated follicles was observed in cultures performed using PCL-Randomic materials (55 vs 37% vs 12.5% for PCL-Randomic vs 3D-oil vs PCL-Patterned, respectively; p <0.0004). The greatest follicle diameter increment (74.7±1 vs 70±0.4 vs 60.9±2%, for PCL-Patterned vs 3D-oil vs PCL-Randomic, respectively p <0.0007) and rate of antrum differentiation (87.5% vs 45% and vs 63%, for PCL-Patterned vs 3D-oil vs PCL-Randomic, for both p <0.0001) were observed in PA ovine follicles cultured on PCL-Patterned scaffolds. Furthermore, PCL-Patterned electrospun scaffolds supported a complete functional development of the oocyte compartment. More in detail, the majority of fully grown oocytes isolated from early- antral follicles grown on PCL-Patterned materials reached the metaphase-II stage (MII 80%) at the end of IVM in comparison to the significant lower percentage in 3D-oil (MII 68%, p =0.04) and PCL-Randomic (MII 18%, p <0.0001) protocols, respectively. Limitations, reasons for caution - Wider implications of the findings Tissue engineering scaffold-based approach represents a valid strategy generating a multi-organ in-vitro system, where different compartments may cooperate generating the complexity of paracrine-mechanism controlling early-follicles outcomes. Scaffold topology is essential to control early-follicles development. Indeed, exclusively PCL-Patterned can preserve long-term follicle 3D-microarchitecture supporting in-vitro oogenesis up to a complete meiotic-competence-acquisition. Trial registration number not applicabl

Meiotic maturation of incompetent prepubertal sheep oocytes is induced by paracrine factor(s) released by gonadotropin-stimulated oocyte-cumulus cell complexes and involves mitogen-activated protein kinase activation.

In this study, sheep oocyte-cumulus cell complexes (OCC) derived from medium (M) antral follicles (M-OCC) were in vitro matured alone or in coculture with OCC derived from small (S) antral follicles (S-OCC) to investigate the contribution of cumulus cells (CC) and oocytes to the process of oocyte meiotic maturation and cumulus expansion (CE). Experiments were conducted with or without gonadotropins (FSH/LH). Regardless of culture conditions, about 12% of S-oocytes reached the metaphase II stage, and S-CC showed a low degree of CE. In contrast, both maturational processes were significantly stimulated by gonadotropins in M-OCC. However, about 48% of S-oocytes progressed to metaphase II, and S-CC expanded after coculture with gonadotropin-stimulated M-OCC and M-CC but not with mural granulosa cells. Both maturational processes were inhibited when S-OCC were cocultured with M-denuded oocytes, or when S-denuded oocytes were cocultured with M-CC. The capacity of these paracrine factor(s) to activate the MAPK pathway in somatic and germ cells of S-complexes was investigated. It was found that MAPK kinase/MAPK phosphorylation levels in M-OCC but not in S-OCC were significantly increased by gonadotropins, first inCCand later in the oocytes. Kinase phosphorylations were activated only in S-oocytes cocultured with M-OCC or M-CC. These results demonstrate that soluble factors specifically produced by M-CC are capable to induce meiotic maturation and CE in S-complexes by acting via CC. These factors can induce MAPK activation only in S-oocytes, whose meiotic arrest could be due to the inability of surrounding CC to respond to gonadotropin stimulation.[...

Esperança de vida ao nascer: impacto das variações na mortalidade por idade e causas de morte no Município de Campinas, São Paulo, Brasil Life expectancy at birth: impact of variation in mortality by age group and cause of death in Campinas, São Paulo State, Brazil

O objetivo do estudo foi examinar o impacto das mudanças na mortalidade por idades e causas de morte sobre o aumento da esperança de vida ao nascer no Município de Campinas, São Paulo, Brasil, entre 1991, 2000 e 2005. Foram construídas tábuas de vida. O método de Pollard foi aplicado para estimar as contribuições das idades e causas de morte na variação da longevidade. O grupo etário de 0-1 ano foi o que mais contribuiu com o aumento da vida média masculina (31,1%) e feminina (22,9%) em 1991/2000. Entre 2000 e 2005, as idades de 15-44 anos responderam por 79% do ganho masculino. A maior contribuição entre 1991 e 2000 foi gerada pelas doenças cardiovasculares (66,1% entre os homens e 43,5% entre as mulheres). As causas externas subtraíram 1,1 ano entre os homens. Entre 2000 e 2005, com a queda da mortalidade por estas causas, a esperança de vida masculina aumentou em 2,3 anos. As neoplasias provocaram redução de 0,11 ano para homens e 0,15 ano para mulheres. Estes resultados podem auxiliar na orientação de políticas públicas de saúde para redução da mortalidade e aumento da esperança de vida ao nascer.<br>This study investigated the impact of variation in mortality by age group and cause of death on gains in life expectancy at birth in the city of Campinas, São Paulo State, Brazil, in 1991, 2000, and 2005. Life tables were constructed. Pollard's method was used to estimate the contributions by age group and cause of death on gains in life expectancy. In 1991-2000, the age group that most contributed was 0-1 year (31.1% for males and 22.9% for females). In 2000-2005, 79% of the gain for males was the result of mortality improvements in the 15-44-year bracket. Cardiovascular diseases made the largest contribution in 1991-2000 (66.1% for males and 43.5% for females). A loss in longevity was seen in men (1.1 year) resulting from increased mortality from external causes. In 2000-2005, the substantial gain (2.3 year) in male life expectancy was due to a reduction in mortality from external causes. Neoplasms had a negative effect on the gain (0.11 year for males and 0.15 for females). These findings should help support public health policies to reduce mortality risks and increase life expectancy

Big Data Analytics for Earth Sciences: the EarthServer approach

Big Data Analytics is an emerging field since massive storage and computing capabilities have been made available by advanced e-infrastructures. Earth and Environmental sciences are likely to benefit from Big Data Analytics techniques supporting the processing of the large number of Earth Observation datasets currently acquired and generated through observations and simulations. However, Earth Science data and applications present specificities in terms of relevance of the geospatial information, wide heterogeneity of data models and formats, and complexity of processing. Therefore, Big Earth Data Analytics requires specifically tailored techniques and tools. The EarthServer Big Earth Data Analytics engine offers a solution for coverage-type datasets, built around a high performance array database technology, and the adoption and enhancement of standards for service interaction (OGC WCS and WCPS). The EarthServer solution, led by the collection of requirements from scientific communities and international initiatives, provides a holistic approach that ranges from query languages and scalability up to mobile access and visualization. The result is demonstrated and validated through the development of lighthouse applications in the Marine, Geology, Atmospheric, Planetary and Cryospheric science domains

Chemical differentiation, cold storage and remobilization of magma in the Earth's crust

The formation, storage and chemical differentiation of magma in the Earth’s crust is of fundamental importance in igneous geology and volcanology. Recent data are challenging the high-melt-fraction ‘magma chamber’ paradigm that has underpinned models of crustal magmatism for over a century, suggesting instead that magma is normally stored in low-melt-fraction ‘mush reservoirs’1,2,3,4,5,6,7,8,9. A mush reservoir comprises a porous and permeable framework of closely packed crystals with melt present in the pore space1,10. However, many common features of crustal magmatism have not yet been explained by either the ‘chamber’ or ‘mush reservoir’ concepts1,11. Here we show that reactive melt flow is a critical, but hitherto neglected, process in crustal mush reservoirs, caused by buoyant melt percolating upwards through, and reacting with, the crystals10. Reactive melt flow in mush reservoirs produces the low-crystallinity, chemically differentiated (silicic) magmas that ascend to form shallower intrusions or erupt to the surface11,12,13. These magmas can host much older crystals, stored at low and even sub-solidus temperatures, consistent with crystal chemistry data6,7,8,9. Changes in local bulk composition caused by reactive melt flow, rather than large increases in temperature, produce the rapid increase in melt fraction that remobilizes these cool- or cold-stored crystals. Reactive flow can also produce bimodality in magma compositions sourced from mid- to lower-crustal reservoirs14,15. Trace-element profiles generated by reactive flow are similar to those observed in a well studied reservoir now exposed at the surface16. We propose that magma storage and differentiation primarily occurs by reactive melt flow in long-lived mush reservoirs, rather than by the commonly invoked process of fractional crystallization in magma chambers[14]