The timing of puberty is highly sensitive to environmental factors, including endocrine disruptors. Among them, bisphenol A (BPA) has been previously analyzed as potential modifier of puberty. Yet, ...disparate results have been reported, with BPA advancing, delaying, or being neutral in its effects on puberty onset. Likewise, mechanistic analyses addressing the central and peripheral actions/targets of BPA at puberty remain incomplete and conflictive.
We aimed to provide a comprehensive characterization of the impact of early BPA exposures, especially at low, real-life doses, on the postnatal development of hypothalamic Kiss1/NKB neurons, and its functional consequences on female pubertal maturation.
Pregnant CD1 female mice were orally administered BPA at 5, 10, or
body weight (BW)/d from gestational day 11 to postnatal day 8 (PND8). Vaginal opening, as an external marker of puberty onset, was monitored daily from PND19 to PND30 in the female offspring. Blood and brain samples were collected at PND12, 15, 18, 21, and 30 for measuring circulating levels of gonadotropins and analyzing the hypothalamic expression of Kiss1/kisspeptin and NKB.
Perinatal exposure to BPA, in a range of doses largely below the no observed adverse effect level (NOAEL;
BW/d, according to the FDA), was associated with pubertal differences in the female progeny compared with those exposed to vehicle alone, with an earlier age of vaginal opening but consistently lower levels of circulating luteinizing hormone. Mice treated with BPA exhibited a persistent, but divergent, impairment of Kiss1 neuronal maturation, with more kisspeptin cells in the rostral (RP3V) hypothalamus but consistently fewer kisspeptin neurons in the arcuate nucleus (ARC). Detailed quantitative analysis of the ARC population, essential for pubertal development, revealed that mice treated with BPA had persistently lower Kiss1 expression during (pre)pubertal maturation, which was associated with lower Tac2 (encoding NKB) levels, even at low doses (
BW/d), in the range of the tolerable daily intake (TDI), recently updated by the European Food Safety Authority.
Our data attest to the consistent, but divergent, effects of gestational exposures to low concentrations of BPA, via the oral route, on phenotypic and neuroendocrine markers of puberty in female mice, with an unambiguous impact on the developmental maturation not only of Kiss1, but also of the NKB system, both essential regulators of puberty onset. https://doi.org/10.1289/EHP5570.
Purpose To redesign the spare parts (MRO) inventory management at Company XYZ's warehouse, considering the conditions after the COVID-19 pandemic.Design/methodology/approach To address this research ...project, the authors integrated three methodologies: action research, Lean Six Sigma (DMAIC) and Cross Industry Standard Process for Data Mining. These methodologies integrated the Lean Six Sigma (LSS) 4.0 framework applied in this project.Findings The spare parts inventory value was reduced by 15%, and inventory turnover increased by 120% without negatively impacting the internal service level.Practical implications Practitioners leading or participating in continuous improvement projects (CIPs) should consider data quality (data available and data trustworthiness), problem-solving approach and target area involvement to achieve CIP goals. Otherwise, the LSS 4.0 could fail or extend its duration by several weeks or months.Originality/value This project shows the importance of controlling a target area before deciding to conduct a LSS 4.0 project. To address this problem, the LSS 4.0 team implemented 5S during the measure phase of the DMAIC. Also, this project offers significant practitioner and theoretical contributions to the body of knowledge about LSS 4.0.
The photoinduced excited‐state dynamics of the keto and enol forms of cytosine have been investigated by using ab initio surface‐hopping to gain an understanding of the outcome of molecular beam ...femtosecond pump‐probe photoionisation spectroscopy experiments. Both singlet and triplet states were included in the dynamics. The results show that triplet states play a significant role in the relaxation of the keto tautomer, whereas they are less important in the enol tautomer. In both forms, the T1 state minimum was found to be too low in energy to be detected in standard photoionisation spectroscopy experiments and therefore experimental decay times should arise from simultaneous relaxation to the ground state and additional intersystem crossing followed by internal conversion to the T1 state. In agreement with available experimental lifetimes, we observed three decay constants of 7, 270 and 1900 fs, the first two coming from the keto tautomer and the third from the enol form. Deactivation of the enol tautomer is due to internal conversion to the ground state through two ethylenic‐type S1/S0 conical intersections.
Alcohol makes you slow…︁ The keto makes you ultrafast! The results of ab initio molecular dynamics studies on cytosine are presented, including for the first time the enol tautomer, which shows a significantly slower relaxation mechanism than the keto form (see picture). The simulations include intersystem crossing to the triplet states, which play an important role in the keto form, but seem to be less accessible in the enol tautomer.
Antimalarial drug resistance compels the quest for new compounds that target alternative pathways to current drugs. The Plasmodium cyclic GMP-dependent protein kinase (PKG) has essential functions in ...all of the major life cycle developmental stages. An imidazopyridine PKG inhibitor scaffold was previously shown to clear P. falciparum infection in a rodent model in vivo and blocked transmission to mosquitoes providing proof of concept for this target. To find new classes of PKG inhibitors to serve as alternative chemical starting points, we performed a high-throughput screen of the GSK Full Diversity Collection using recombinant P. falciparum PKG. We developed a robust enzymatic assay in a 1536-well plate format. Promising compounds were then tested for activity against P. falciparum asexual blood stage growth, selectivity and cytotoxicity. By using a scoring system we selected the 66 most promising PKG inhibitors (comprising nine clusters and seven singletons). Among these, thiazoles were the most potent scaffold with mid-nanomolar activity on P. falciparum blood stage and gamete development. Using Kinobeads profiling we identified additional P. falciparum protein kinases targeted by the thiazoles that mediate a faster speed of the kill than PKG-selective compounds. This scaffold represents a promising starting point to develop a new antimalarial.
High-throughput sequencing of full-length transcripts using long reads has paved the way for the discovery of thousands of novel transcripts, even in well-annotated mammalian species. The advances in ...sequencing technology have created a need for studies and tools that can characterize these novel variants. Here, we present SQANTI, an automated pipeline for the classification of long-read transcripts that can assess the quality of data and the preprocessing pipeline using 47 unique descriptors. We apply SQANTI to a neuronal mouse transcriptome using Pacific Biosciences (PacBio) long reads and illustrate how the tool is effective in characterizing and describing the composition of the full-length transcriptome. We perform extensive evaluation of ToFU PacBio transcripts by PCR to reveal that an important number of the novel transcripts are technical artifacts of the sequencing approach and that SQANTI quality descriptors can be used to engineer a filtering strategy to remove them. Most novel transcripts in this curated transcriptome are novel combinations of existing splice sites, resulting more frequently in novel ORFs than novel UTRs, and are enriched in both general metabolic and neural-specific functions. We show that these new transcripts have a major impact in the correct quantification of transcript levels by state-of-the-art short-read-based quantification algorithms. By comparing our iso-transcriptome with public proteomics databases, we find that alternative isoforms are elusive to proteogenomics detection. SQANTI allows the user to maximize the analytical outcome of long-read technologies by providing the tools to deliver quality-evaluated and curated full-length transcriptomes.
The role of alternative splicing is one of the great unanswered questions in cellular biology. There is strong evidence for alternative splicing at the transcript level, and transcriptomics ...experiments show that many splice events are tissue specific. It has been suggested that alternative splicing evolved in order to remodel tissue-specific protein-protein networks. Here we investigated the evidence for tissue-specific splicing among splice isoforms detected in a large-scale proteomics analysis. Although the data supporting alternative splicing is limited at the protein level, clear patterns emerged among the small numbers of alternative splice events that we could detect in the proteomics data. More than a third of these splice events were tissue-specific and most were ancient: over 95% of splice events that were tissue-specific in both proteomics and RNAseq analyses evolved prior to the ancestors of lobe-finned fish, at least 400 million years ago. By way of contrast, three in four alternative exons in the human gene set arose in the primate lineage, so our results cannot be extrapolated to the whole genome. Tissue-specific alternative protein forms in the proteomics analysis were particularly abundant in nervous and muscle tissues and their genes had roles related to the cytoskeleton and either the structure of muscle fibres or cell-cell connections. Our results suggest that this conserved tissue-specific alternative splicing may have played a role in the development of the vertebrate brain and heart.
•An artificial vision algorithm is proposed to reach the MPP under partial shading conditions.•The artificial vision algorithm using a webcam provides the shadow irradiance.•This method also provides ...the voltage that supplies de MPP.•The MPP is tracked using a backstepping controller for the DC/DC converter.
Maximum power point tracking (MPPT) algorithms should track and extract the maximum power from photovoltaic (PV) systems under any environmental conditions. Most of conventional MPPT methods are able to reach the maximum point when there is only one peak in the P–V characteristic curve but they fail when the solar cells are affected by partial shading conditions due to the fact that multiple peaks appear in the P–V curve. Thus, a local maximum may be reached instead of the global peak. In this work, a new method to accomplish the maximum power point (MPP) under partial shading conditions using artificial vision is presented. The artificial vision uses a webcam to identify in real time the shadow irradiance and provide the reference voltage that supplies the maximum power, regardless of the number of peaks that the P–V curve presents. Then, the reference voltage is used by a robust and non-linear control, the backstepping controller, to regulate the DC/DC converter input voltage and to guarantee the PV modules maximum energy extraction. Experimental tests carried out outdoor validate the proposed method, obtaining a MPP tracking efficiency that ranges from 98.1% to 99.6%.
Lipid droplets (LDs) are the major lipid storage organelles of eukaryotic cells and a source of nutrients for intracellular pathogens. We demonstrate that mammalian LDs are endowed with a ...protein-mediated antimicrobial capacity, which is up-regulated by danger signals. In response to lipopolysaccharide (LPS), multiple host defense proteins, including interferon-inducible guanosine triphosphatases and the antimicrobial cathelicidin, assemble into complex clusters on LDs. LPS additionally promotes the physical and functional uncoupling of LDs from mitochondria, reducing fatty acid metabolism while increasing LD-bacterial contacts. Thus, LDs actively participate in mammalian innate immunity at two levels: They are both cell-autonomous organelles that organize and use immune proteins to kill intracellular pathogens as well as central players in the local and systemic metabolic adaptation to infection.
Fretting fatigue damage is a typical material damage arising in mechanical components. In such a damage condition, surface cracks can initiate at the surface contact zone, and under a fluctuating ...loading they can grow up to the complete failure of the component, resulting in a catastrophic failure.
In order to prevent/mitigate fretting fatigue damage, shot peening is widely used in mechanical components. The improvement due to shot peening is related to different factors, but the main one is the compressive residual stress field produced by such a treatment in a region near the contact surface. Unfortunately, both the uncertainty in measurements and stability (relaxation) of residual stress field, make shot peening effects difficult to be included in models aimed to the estimation of fatigue life. Due to this lack in the literature, especially relatively to the fretting fatigue field, in the present paper a new fretting fatigue model, that is able to consider shot peening residual stress and its relaxation, is presented. A 3D elastic-plastic finite element model, which incorporates the shot peening residual stress and its relaxations, is employed. The results obtained, in terms of stress field, are used as input data of a multiaxial stress-based criterion to estimate both fatigue lifetime and crack initiation direction.
•A new fretting fatigue model able to consider shot peening residual stress is presented.•The residual stability of the shot peening residual stress during fretting fatigue life is considered.•Results are compared with those coming from an experimental campaign performed on Al 7075-T651 specimens.•The model estimates well the fatigue life and crack initiation direction.
Abstract Objective Puberty is a key developmental phenomenon highly sensitive to metabolic modulation. Worrying trends of changes in the timing of puberty have been reported in humans. These might be ...linked to the escalating prevalence of childhood obesity and could have deleterious impacts on later (cardio-metabolic) health, but their underlying mechanisms remain unsolved. The neuropeptide α-MSH, made by POMC neurons, plays a key role in energy homeostasis by mediating the actions of leptin and likely participates in the control of reproduction. However, its role in the metabolic regulation of puberty and interplay with kisspeptin, an essential puberty-regulating neuropeptide encoded by Kiss1 , remain largely unknown. We aim here to unveil the potential contribution of central α-MSH signaling in the metabolic control of puberty by addressing its role in mediating the pubertal effects of leptin and its potential interaction with kisspeptin. Methods Using wild type and genetically modified rodent models, we implemented pharmacological studies, expression analyses, electrophysiological recordings, and virogenetic approaches involving DREADD technology to selectively inhibit Kiss1 neurons, in order to interrogate the physiological role of a putative leptin→α-MSH→kisspeptin pathway in the metabolic control of puberty. Results Stimulation of central α-MSH signaling robustly activated the reproductive axis in pubertal rats, whereas chronic inhibition of melanocortin receptors MC3/4R, delayed puberty, and prevented the permissive effect of leptin on puberty onset. Central blockade of MC3/4R or genetic elimination of kisspeptin receptors from POMC neurons did not affect kisspeptin effects. Conversely, congenital ablation of kisspeptin receptors or inducible, DREADD-mediated inhibition of arcuate nucleus (ARC) Kiss1 neurons resulted in markedly attenuated gonadotropic responses to MC3/4R activation. Furthermore, close appositions were observed between POMC fibers and ARC Kiss1 neurons while blockade of α-MSH signaling suppressed Kiss1 expression in the ARC of pubertal rats. Conclusions Our physiological, virogenetic, and functional genomic studies document a novel α-MSH→kisspeptin→GnRH neuronal signaling pathway involved in transmitting the permissive effects of leptin on pubertal maturation, which is relevant for the metabolic (and, eventually, pharmacological) regulation of puberty onset.
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