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Abstract Background Recently, an effective testis culture method using a gas-liquid interphase, capable of differentiate male germ cells from neonatal spermatogonia to spermatozoa has been developed. Nevertheless, this methodology needs deep analyses that allow future experimental approaches in basic, pathologic and/or reprotoxicologic studies. Because of this, we characterized at cellular and molecular levels the entire in vitro spermatogenic progression, in order to understand and evaluate the characteristics that define the spermatogenic process in ex vivo cultured testes compared to the in vivo development. Methods Testicular explants of CD1 mice aged 6 and 10 days post-partum were respectively cultured during 55 and 89 days. Cytological and molecular approaches were performed, analyzing germ cell proportion at different time culture points, meiotic markers immunodetecting synaptonemal complex protein SYCP3 by immunocytochemistry and the relative expression of different marker genes along the differentiation process by Reverse Transcription - quantitative Polymerase Chain Reaction. In addition, microRNA and piwi-interactingRNA profiles were also evaluated by Next Generation Sequencing and bioinformatic approaches. Results The method promoted and maintained the spermatogenic process during 89 days. At a cytological level we detected spermatogenic development delays of cultured explants compared to the natural in vivo process. The expression of different spermatogenic stages gene markers correlated with the proportion of different cell types detected in the cytological preparations. Conclusions In vitro progression analysis of the different spermatogenic cell types, from both 6.5 dpp and 10.5 dpp testes explants, has revealed a relative delay in relation to in vivo process. The expression of the genes studied as biomarkers correlates with the cytologically and functional detected progression and differential expression identified in vivo. After a first analysis of deep sequencing data it has been observed that as long as cultures progress, the proportion of microRNAs declined respect to piwi-interactingRNAs levels that increased, showing a similar propensity than which happens in in vivo spermatogenesis. Our study allows to improve and potentially to control the ex vivo spermatogenesis development, opening new perspectives in the reproductive biology fields including male fertility.

Abstract A new mobility particle analyzer, which has been termed Inverted Drift Tube, has been modeled analytically as well as numerically and proven to be a very capable instrument. The basis for the new design have been the shortcomings of the previous ion mobility spectrometers, in particular (a) diffusional broadening which leads to degradation of instrument resolution and (b) inadequate low and fixed resolution (not mobility dependent) for large sizes. To overcome the diffusional broadening and have a mobility based resolution, the IDT uses two varying controllable opposite forces, a flow of gas with velocity v gas , and a linearly increasing electric field that opposes the movement. A new parameter, the separation ratio Λ = v drift /v gas , is employed to determine the best possible separation for a given set of nanoparticles. Due to the system’s need to operate at room pressure, two methods of capturing the ions at the end of the drift tube have been developed, Intermittent Push Flow for a large range of mobilities, and Nearly-Stopping Potential Separation, with very high separation but limited only to a narrow mobility range. A chromatography existing concept of resolving power is used to differentiate between peak resolution in the IDT and acceptable separation between similar mobility sizes.

Abstract Humans are environmentally exposed not only to single endocrine-disrupting chemicals (EDCs) but to mixtures that affect their reproductive health. In reproductive tissues, microRNAs (miRNAs) are emerging as key targets of EDCs. Here, we analysed changes in the testis “miRNome” (and their biogenesis mechanism) in chronically exposed adult mice to a cocktail of five EDCs containing 0.3 mg/kg-body weight (BW)/day of each phthalate (DEHP, DBP, BBP) and 0.05 mg/kg-BW/day of each alkylphenol (NP, OP), from conception to adulthood. The testis “miRNome” was characterised using next-generation sequencing (NGS). Expression levels of genes involved in miRNA biogenesis were measured by RT-qPCR, as well as several physiological and cytological parameters. We found two up-regulated, and eight down-regulated miRNAs and thirty-six differentially expressed isomiRs along with an over-expression of Drosha, Adar and Zcchc11. A significant decrease of intratesticular estradiol but not testosterone was detected. Functional analysis showed altered spermatogenesis, germ cell apoptosis and negative correlation of miR-18a-5p with Nr1h2 involved in the deregulation of the steroidogenesis pathway. Here, we present the first association between miRNA/isomiRs deregulation, their mechanisms of biogenesis and histopathological and hormonal alterations in testes of adult mice exposed to a mixture of low-dose EDCs, which can play a role in male infertility.

Oscillatory systems arise in the different biological and medical fields. Mathematical and statistical approaches are fundamental to deal with these processes. The Frequency Modulated Mobiüs approach (FMM), reviewed in this paper, is one of these approaches. Little known as it has been recently developed, it solves a variety of exciting questions with real data; some of them, such as the decomposition of the signal into components and their multiple uses, are of general application, others are specific. Among the exciting specific applications is the automatic interpretation of the electrocardiogram signal. In this paper, a summary of the theoretical, statistical and computational properties of the FMM approach are revised. Additionally, as a novelty, the FMM approach’s usefulness for the analysis of blood pressure signals is shown. For the latter, a new robust estimation algorithm is proposed using FMM models with restrictions. The paper ends with a view about challenges for the future.

The amount of sludge produced in wastewater treatment plants (WWTPs) has increased over the years, and the methods used to reduce this waste, such as incineration, agricultural use, or disposal in landfills, cause problems of secondary pollution. For this reason, it is necessary to find sustainable and low-cost solutions to manage this waste. Additionally, emerging and priority pollutants are attracting attention from the scientific community as they can generate health problems due to inadequate removal in conventional WWTPs. In this work, a pharmaceutical industry sludge was used as a precursor in the synthesis of four activated carbons (ACs) using different activating agents (ZnCl2, FeCl3∙6H2O, Fe(NO3)3∙9H2O, and Fe(SO4)3∙H2O), to be used for the removal by adsorption of three neonicotinoid pesticides included in latest EU Watch List (Decision 2018/840): acetamiprid (ACT), thiamethoxam (THM), and imidacloprid (IMD). The prepared ACs showed micro–mesoporous properties, obtaining relatively slow adsorption kinetics to reach equilibrium, but despite this, high values of adsorption capacity (qe) were obtained. For example, for AC-ZnCl2 (SBET = 558 m2/g), high adsorption capacities of qe = 128.9, 126.8, and 166.1 mg/g for ACT, THM, and IMD, respectively, were found. In most cases, the adsorption isotherms showed a multilayer profile, indicating an important contribution of the mesoporosity of the activated carbons in the adsorption process.