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Abiotic D-proteins that selectively bind to natural L-proteins have gained significant biotechnological interest. However, the underlying structural principles governing such heterochiral protein-protein interactions remain largely unknown. In this study, we present the de novo design of D-proteins consisting of 50-65 residues, aiming to target specific surface regions of L-proteins or L-peptides. Our designer D-protein binders exhibit nanomolar affinity toward an artificial L-peptide, as well as two naturally occurring proteins of therapeutic significance: the D5 domain of human tropomyosin receptor kinase A (TrkA) and human interleukin-6 (IL-6). Notably, these D-protein binders demonstrate high enantiomeric specificity and target specificity. In cell-based experiments, designer D-protein binders effectively inhibited the downstream signaling of TrkA and IL-6 with high potency. Moreover, these binders exhibited remarkable thermal stability and resistance to protease degradation. Crystal structure of the designed heterochiral D-protein-L-peptide complex, obtained at a resolution of 2.0 Å, closely resembled the design model, indicating that the computational method employed is highly accurate. Furthermore, the crystal structure provides valuable information regarding the interactions between helical L-peptides and D-proteins, particularly elucidating a novel mode of heterochiral helix-helix interactions. Leveraging the design of D-proteins specifically targeting L-peptides or L-proteins opens up avenues for systematic exploration of the mirror-image protein universe, paving the way for a diverse range of applications.
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PM2.5 exposure has been found to cause gut dysbiosis and impair glucose homeostasis in human and animals, yet their underlying biological connection remain unclear. In the present study, we aim to investigate the biological significance of gut microbiota in PM2.5-induced glucose metabolic abnormalities. Our results showed that microbiota depletion by antibiotics treatment significantly alleviated PM2.5-induced glucose intolerance and insulin resistance, as indicated by the intraperitoneal glucose tolerance test, glucose-induced insulin secretion, insulin tolerance test, insulin-induced phosphorylation levels of Akt and GSK-3ß in insulin sensitive tissues. In addition, faecal microbiota transplantation (FMT) from PM2.5-exposed donor mice successfully remodeled the glucose metabolism abnormalities in recipient mice, while the transplantation of autoclaved faecal materials did not. Faecal microbiota analysis demonstrated that the composition and alpha diversity of the gut bacterial community were altered by PM2.5 exposure and in FMT recipient mice. Furthermore, short-chain fatty acids levels analysis showed that the circulating acetate was significantly decreased in PM2.5-exposed donor and FMT recipient mice, and supplementation of sodium acetate for 3 months successfully improved the glucose metabolism abnormalities induced by PM2.5 exposure. These results indicate that manipulating gut microbiota or its metabolites could be a potential strategy for preventing the adverse health effects of ambient PM2.5.
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Ácidos Graxos Voláteis , Microbioma Gastrointestinal , Resistência à Insulina , Material Particulado , Animais , Microbioma Gastrointestinal/efeitos dos fármacos , Ácidos Graxos Voláteis/metabolismo , Material Particulado/toxicidade , Masculino , Camundongos Endogâmicos C57BL , Transplante de Microbiota Fecal , Glucose/metabolismo , Intolerância à Glucose/metabolismo , Poluentes Atmosféricos/toxicidade , Camundongos , Antibacterianos/farmacologia , Disbiose/induzido quimicamente , Disbiose/metabolismo , Fezes/microbiologia , Acetato de Sódio/farmacologia , Teste de Tolerância a Glucose , Insulina/metabolismo , Insulina/sangueRESUMO
Stress is known to disrupt the intestinal barrier and induce intestinal dysfunction. A critical role for gonadotropin inhibitory hormone (GnIH) in stress has emerged. However, whether GnIH mediates stress-induced intestinal dysfunction remains unknown. The present study explored this question through in vivo and in vitro experiments in hens. Our in vivo experiments showed that continuous intraperitoneal injection of GnIH not only significantly increased the concentration of stress hormones in serum, but also significantly elevated the mRNA expression of glucocorticoid receptor (GR) in the duodenum and jejunum. Moreover, morphological and molecular analyses revealed that GnIH disrupted the physical and chemical barriers of the intestine and dramatically increased inflammatory factor levels in the intestine and serum of hens. Interestingly, the microbiomics results showed that GnIH altered the structure and composition of the gut flora in the cecum, revealing an increased abundance of harmful intestinal bacteria such as Desulfovibrionaceae. Similar results were found in in vitro studies in which the GnIH-induced intestinal mucosal barrier was disrupted, and inflammation increased in jejunal explants, although no significant difference was found in the expression of GR between the control and GnIH groups. Our results demonstrated that GnIH not only directly damaged intestinal barriers and elevated intestinal inflammation but also mediated stress and microflora imbalance-induced intestinal function disorder, suggesting that GnIH is a potential therapeutic target for gut dysfunction, stress-induced intestinal function disorder, and inflammatory bowel disease in animals and humans.
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Galinhas , Microbioma Gastrointestinal , Estresse Fisiológico , Animais , Galinhas/fisiologia , Feminino , Microbioma Gastrointestinal/fisiologia , Hormônios Hipotalâmicos/metabolismo , Hormônios Hipotalâmicos/genética , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/fisiopatologia , Proteínas Aviárias/metabolismo , Proteínas Aviárias/genética , Enteropatias/veterinária , Enteropatias/microbiologiaRESUMO
INTRODUCTION: Since the discovery of gonadotropin-inhibitory hormone (GnIH), it has been found to play a critical role in reproduction in vertebrates. Recently, a regulatory role of GnIH in appetite and energy metabolism has emerged, although its precise physiological mechanisms remain unknown. METHODS: Thus, the present study evaluated the effects of a single or long-term intraperitoneal GnIH treatment on the food intake, weight, and glucolipid metabolism of chickens, as well as investigating the possible neuroendocrinology factors and mechanisms involved in GnIH-induced obesity and glucolipid metabolism disorder. RESULTS: Our results show that the intraperitoneal administration of GnIH to chickens resulted in a marked body mass increase, hyperlipidemia, hyperglycemia, and glucose intolerance. Subsequently, the results of metabolomics studies and the pharmacological inhibition of the 5-HT2C receptor revealed that blocking the 5-HT2C receptor reinforced the effects of GnIH on food intake, body weight, and blood glucose and lipid levels, resulting in even worse cases of GnIH-induced hyperglycemia, hyperlipidemia, and hepatic lipid deposition. This suggests that, via the 5-HT2C receptor, peripheral 5-HT may act as a negative feedback regulator to interplay with GnIH and jointly control energy balance homeostasis in chickens. DISCUSSION: Our present study provides evidence of cross-talk between GnIH and 5-HT in food intake and energy metabolism at the in vivo pharmacological level, and it proposes a molecular basis for these interactions, suggesting that functional interactions between GnIH and 5-HT may open new avenues for understanding the mechanism of the neuroendocrine network involved in appetite and energy metabolism, as well as providing a new therapeutic strategy to prevent obesity, diabetes, and metabolic disorders.
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Galinhas , Metabolismo Energético , Comportamento Alimentar , Receptor 5-HT2C de Serotonina , Serotonina , Animais , Metabolismo Energético/efeitos dos fármacos , Receptor 5-HT2C de Serotonina/metabolismo , Serotonina/metabolismo , Comportamento Alimentar/efeitos dos fármacos , Comportamento Alimentar/fisiologia , Hormônios Hipotalâmicos/metabolismo , Masculino , Glicemia/metabolismo , Glicemia/efeitos dos fármacos , Hiperlipidemias/metabolismo , Hiperlipidemias/induzido quimicamenteRESUMO
Mirror-image proteins (D-proteins) are useful in biomedical research for purposes such as mirror-image screening for D-peptide drug discovery, but the chemical synthesis of many D-proteins is often low yielding due to the poor solubility or aggregation of their constituent peptide segments. Here, we report a Lys-C protease-cleavable solubilizing tag and its use to synthesize difficult-to-obtain D-proteins. Our tag is easily installed onto multiple amino acids such as DLys, DSer, DThr, and/or the N-terminal amino acid of hydrophobic D-peptides, is impervious to various reaction conditions, such as peptide synthesis, ligation, desulfurization, and transition metal-mediated deprotection, and yet can be completely removed by Lys-C protease under denaturing conditions to give the desired D-protein. The efficacy and practicality of the new method were exemplified in the synthesis of two challenging D-proteins: D-enantiomers of programmed cell death protein 1 IgV domain and SARS-CoV-2 envelope protein, in high yield. This work demonstrates that the enzymatic cleavage of solubilizing tags under denaturing conditions is feasible, thus paving the way for the production of more D-proteins.
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Peptídeos , Proteínas , Proteínas/química , Peptídeos/química , Aminoácidos/química , Técnicas de Química Sintética/métodos , Peptídeo Hidrolases , EndopeptidasesRESUMO
D-peptide ligands can be screened for therapeutic potency and enzymatic stability using synthetic mirror-image proteins (D-proteins), but efficient acquisition of these D-proteins can be hampered by the need to accomplish their in vitro folding, which often requires the formation of correctly linked disulfide bonds. Here, we report the finding that temporary installation of natural O-linked-ß-N-acetyl-D-glucosamine (O-GlcNAc) groups onto selected D-serine or D-threonine residues of the synthetic disulfide-bonded D-proteins can facilitate their folding in vitro, and that the natural glycosyl groups can be completely removed from the folded D-proteins to afford the desired chirally inverted D-protein targets using naturally occurring O-GlcNAcase. This approach enabled the efficient chemical syntheses of several important but difficult-to-fold D-proteins incorporating disulfide bonds including the mirror-image tumor necrosis factor alpha (D-TNFα) homotrimer and the mirror-image receptor-binding domain of the Omicron spike protein (D-RBD). Our work establishes the use of O-GlcNAc to facilitate D-protein synthesis and folding and proves that D-proteins bearing O-GlcNAc can be good substrates for naturally occurring O-GlcNAcase.
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Acetilglucosaminidase , Proteínas , Peptídeos , Polissacarídeos , GlucosaminaRESUMO
Ambient particles severely threaten human health worldwide. Compared to larger particles, ultrafine particles (UFPs) are highly concentrated in ambient environments, have a larger specific surface area, and are retained for a longer time in the lung. Recent studies have found that they can be transported into various extra-pulmonary organs by crossing the air-blood barrier (ABB). Therefore, to understand the adverse effects of UFPs, it is crucial to thoroughly investigate their bio-distribution and clearance pathways in vivo after inhalation, as well as their toxicological mechanisms. This review highlights emerging evidence on the bio-distribution of UFPs in pulmonary and extra-pulmonary organs. It explores how UFPs penetrate the ABB, the blood-brain barrier (BBB), and the placental barrier (PB) and subsequently undergo clearance by the liver, kidney, or intestine. In addition, the potential underlying toxicological mechanisms of UFPs are summarized, providing fundamental insights into how UFPs induce adverse health effects.
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Lily (Lilium spp.) is a popular ornamental plant. Traditional genetic transformation methods have low efficiency in lily, thus development of a high-efficiency genetic transformation system is important. In this study, a novel transient transformation method involving pollen magnetofection was established and optimized pollen viability, and exogenous gene expression in magnetofected pollen and that of different germplasm were assessed. The highest germination percentage of Lilium regale pollen was 85.73% in medium containing 100 g/L sucrose, 61.5 mg/L H3BO3, and 91.5 mg/L CaCl2. A 1:4 ratio of nanomagnetic beads to DNA plasmid and transformation time of 0.5 h realized the highest transformation efficiency (88.32%). The GFP activity in transformed pollen averaged 69.66%, while that of the control pollen was 0.00%. In contrast to the control, transgenic seedlings obtained by pollination with magnetofected pollen showed strong positive GUS activity with 56.34% transformation efficiency. Among the lily germplasm tested, 'Sweet Surrender' and L. leucanthum had the highest transformation efficiency (85.80% and 54.47%), whereas L. davidii var. willmottiae was not successfully transformed. Transformation efficiency was positively correlated with pollen equatorial diameter and negatively correlated with polar axis/equatorial diameter ratio. The results suggest that pollen magnetofection-mediated transformation can be applied in Lilium but might have species or cultivar specificity.
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Lilium , Lilium/genética , Lilium/metabolismo , Pólen/genética , Pólen/metabolismo , Proteínas de Plantas/genéticaRESUMO
Aryl sulfides are common and ubiquitous motifs in natural products and pharmaceuticals. Presented herein is the first example of the synthesis of diaryl sulfide derivatives via dehydroaromatization under simple basic conditions. Dehydroaromatization reactions between indolines or cyclohexanones with aryl thiols are performed in an environmentally benign manner by the use of air (molecular oxygen) as the oxidant, with producing water as the only byproduct. The methodology provides a simple and practical route to diaryl sulfides with wide functional groups in good to excellent yields. Preliminary mechanistic studies suggest that a radical process is involved in the transformation.
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Previous research has indicated that the cholinergic anti-inflammatory pathway (CAP) can regulate the duration and intensity of inflammatory responses. A wide range of research has demonstrated that PM2.5 exposure may induce various negative health effects via pulmonary and systemic inflammations. To study the potential role of the CAP in mediating PM2.5-induced effects, mice were treated with vagus nerve electrical stimulation (VNS) to activate the CAP before diesel exhaust PM2.5 (DEP) instillation. Analysis of pulmonary and systemic inflammations in mice demonstrated that VNS significantly reduced the inflammatory responses triggered by DEP. Meanwhile, inhibition of the CAP by vagotomy aggravated DEP-induced pulmonary inflammation. The flow cytometry results showed that DEP influenced the CAP by altering the Th cell balance and macrophage polarization in spleen, and in vitro cell co-culture experiments indicated that this DEP-induced change on macrophage polarization may act via the splenic CD4+ T cells. To further confirm the effect of alpha7 nicotinic acetylcholine receptor (α7nAChR) in this pathway, mice were then treated with α7nAChR inhibitor (α-BGT) or agonist (PNU282987). Our results demonstrated that specific activation of α7nAChR with PNU282987 effectively alleviated DEP-induced pulmonary inflammation, while specific inhibition of α7nAChR with α-BGT exacerbated the inflammatory markers. The present study suggests that PM2.5 have an impact on the CAP, and CAP may play a critical function in mediating PM2.5 exposure-induced inflammatory response. AVAILABILITY OF DATA AND MATERIALS: The datasets used and/or analyzed during the present study are available from the corresponding author on reasonable request.
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Pneumonia , Emissões de Veículos , Camundongos , Animais , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Neuroimunomodulação , Inflamação/metabolismo , Material ParticuladoRESUMO
Adsorption-diffusion behaviors of gases (i.e., O2, CO2, and N2) in coal are directly related to the coal spontaneous combustion (CSC), in which the temperature is the key factor affecting the gas migration process in coal. In this work, isothermal adsorption experiments of O2, CO2, and N2 under different temperatures were carried out on bituminous coal and anthracite coal samples at 0.5 MPa, respectively. Based on the free gas density gradient diffusion (FDGD) model, the microchannel diffusion coefficients of different gases at different temperatures were calculated, and the effects from temperature were quantitatively evaluated. The results acquired from the experiment and simulation show that (i) the adsorption capacity of these three gases decreases as the temperature increases, and the adsorption capacity at the same temperature satisfies CO2 > O2 > N2; (ii) the FDGD model is verified to be still applicable at different temperatures, indicating that the adsorption-diffusion behavior of O2, CO2, and N2 in coal particles at different temperatures is still consistent with the FDGD diffusion; (iii) the microchannel diffusion coefficient Km of the three gases gradually increases when the temperature goes up. The present work contributes to the understanding of the gases migration process in the development of CSC.
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Dióxido de Carbono , Carvão Mineral , Adsorção , Temperatura , Gases/análiseRESUMO
Highly bio-compatible organic semiconductors are widely used as biosensors, but their long-term stability can be compromised due to photo-degradation and structural instability. To address this issue, scientists have developed organic semiconductor nanoparticles (OSNs) by incorporating organic semiconductors into a stable framework or self-assembled structure. OSNs have shown excellent performance and can be used as high-resolution biosensors in modern medical and biological research. They have been used for a wide range of applications, such as detecting small biological molecules, nucleic acids, and enzyme levels, as well as vascular imaging, tumor localization, and more. In particular, OSNs can simulate fine particulate matters (PM2.5, indicating particulate matter with an aerodynamic diameter less than or equal to 2.5 µm) and can be used to study the biodistribution, clearance pathways, and health effects of such particles. However, there are still some problems that need to be solved, such as toxicity, metabolic mechanism, and fluorescence intensity. In this review, based on the structure and design strategies of OSNs, we introduce various types of OSNs-based biosensors with functional groups used as biosensors and discuss their applications in both in vitro and in vivo tracking. Finally, we also discuss the design strategies and potential future trends of OSNs-based biosensors. This review provides a theoretical scaffold for the design of high-performance OSNs-based biosensors and highlights important trends and future directions for their development and application.
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Técnicas Biossensoriais , Nanopartículas , Pontos Quânticos , Distribuição Tecidual , Técnicas Biossensoriais/métodos , SemicondutoresRESUMO
Aquatic community dynamics are closely dominated by flow regime and water quality conditions, which are increasingly threatened by dam regulation, water diversion, and nutrition pollution. However, further understanding of the ecological impacts of flow regime and water quality conditions on aquatic multi-population dynamics has rarely been integrated into existing ecological models. To address this issue, a new niche-based metacommunity dynamics model (MDM) is proposed. The MDM aims to simulate the coevolution processes of multiple populations under changing abiotic environments, pioneeringly applied to the mid-lower Han River, China. The quantile regression method was used for the first time to derive ecological niches and competition coefficients of the MDM, which are demonstrated to be reasonable by comparing them with the empirical evidence. Simulation results show that the Nash efficiency coefficients for fish, zooplankton, zoobenthos, and macrophytes are more than 0.64, while the Pearson correlation coefficients for them are no less than 0.71. Overall, the MDM performs effectively in simulating metacommunity dynamics. For all river stations, the average contributions of biological interaction, flow regime effects, and water quality effects to multi-population dynamics are 64%, 21%, and 15%, respectively, suggesting that the population dynamics are dominated by biological interaction. For upstream stations, the fish population is 8%-22% more responsive to flow regime alteration than other populations, while other populations are 9%-26% more responsive to changes in water quality conditions than fish. For downstream stations, flow regime effects on each population account for less than 1% due to more stable hydrological conditions. The innovative contribution of this study lies in proposing a multi-population model to quantify the effects of flow regime and water quality on aquatic community dynamics by incorporating multiple indicators of water quantity, water quality, and biomass. This work has potential for the ecological restoration of rivers at the ecosystem level. This study also highlights the importance of considering threshold and tipping point issues when analyzing the "water quantity-water quality-aquatic ecology" nexus in future works.
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Ecossistema , Qualidade da Água , Animais , Biomassa , Hidrologia , RiosRESUMO
The arylcyclopropane motif as the combination of aryl and cyclopropyl ring systems can be found in an increasing amount of approved and investigational drugs. Herein, we have developed a mild, efficient nickel-catalyzed reductive cross-coupling protocol, featuring a simple Ni(II) precatalyst and a novel picolinamide NN2 pincer ligand. A variety of (hetero)aryl bromides could successfully couple with cyclopropyl bromide to furnish the valued arylcyclopropanes in good to excellent yields. This method is applicable to other alkyl bromides as well. Notably, the reaction is tolerant of a broad range of functionalities including free amines. Furthermore, the synthesis of several significant intermediates of bioactive molecules was achieved in grams, proving the practicability of this method.
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Brometos , Níquel , Ligantes , Estrutura Molecular , CatáliseRESUMO
Epidemiological and animal studies have shown that maternal fine particulate matters (PM2.5) exposure correlates with various adverse pregnancy outcomes such as low birth weight (LBW) of offspring. However, the underlying biological mechanisms have not been fully understood. In this study, female C57Bl/6 J mice were exposed to filtered air (FA) or concentrated ambient PM2.5 (CAP) during pregestational and gestational periods, and metabolomics was performed to analyze the metabolic features in maternal serum and placenta by liquid chromatography-mass spectrometry (LC-MS). The partial least squares discriminate analysis (PLS-DA) displayed evident clustering of FA- and CAP-exposed samples for both maternal serum and placenta. In addition, pathway analysis identified that vitamin digestion and absorption was perturbed in maternal serum, while metabolic pathways including arachidonic acid metabolism, serotonergic synapse, 2-oxocarboxylic acid metabolism and cAMP signaling pathway were perturbed in placenta. Further analysis indicated that CAP exposure influenced the nutrient transportation capacity of placenta, by not only changing the ratios of some critical metabolites in placenta to maternal serum but also significantly altering the expressions of nutrition transporters in placenta. These findings reaffirm the importance of protecting women from PM2.5 exposure, and also advance our understanding of the toxic actions of ambient PM2.5.
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Poluentes Atmosféricos , Exposição Materna , Gravidez , Humanos , Feminino , Camundongos , Animais , Exposição Materna/efeitos adversos , Poluentes Atmosféricos/análise , Material Particulado/análise , Placenta/química , Camundongos Endogâmicos C57BL , HomeostaseRESUMO
Apart from the well-established role of the gonadotropin-inhibitory hormone (GnIH) in the regulation of the reproductive functions, much less is known about the peripheral role of the GnIH and its receptor in the metabolic processes. On account of pig being an excellent model for studies of food intake and obesity in humans, we investigated the peripheral effects of the GnIH on food intake and energy homeostasis and revealed the underlying mechanism(s) in female piglets in vivo. Compared to the vehicle-treated group, intraperitoneally injected GnIH significantly increased the food intake and altered the meal microstructure both in the fasting and ad libitum female piglet. GnIH-triggered hyperphagia induced female piglet obesity and altered islet hormone secretion in the pancreas, accompanied with dyslipidemia and hyperglycemia. Interestingly, GnIH decreased the glucose transport capacity and glycogen synthesis, whereas it increased the gluconeogenesis in the liver, while it also induced an insulin resistance in white adipose tissue (WAT) via inhibiting the activity of AKT-GSK3-ß signaling. In terms of the lipid metabolism, GnIH reduced the oxidation of fatty acids, whereas the elevated fat synthesis ability in the liver and WAT was developed though the inhibited AMPK phosphorylation. Our findings demonstrate that peripheral GnIH could trigger hyperphagia-induced obesity and an associated glycolipid metabolism disorder in female piglets, suggesting that GnIH may act as a potential therapeutic agent for metabolic syndrome, obesity and diabetes.
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Hormônios Hipotalâmicos , Humanos , Animais , Feminino , Suínos , Hormônios Hipotalâmicos/fisiologia , Quinase 3 da Glicogênio Sintase , Gonadotropinas , Hiperfagia , Obesidade/etiologiaRESUMO
Gonadotropin-inhibitory hormone (GnIH) is a reproductive inhibitor and an endogenous orexigenic neuropeptide that may be involved in energy homeostasis and reproduction. However, whether GnIH is a molecular signal link of metabolism and the reproductive system, and thus, regulates reproductive activity as a function of the energy state, is still unknown. In the present study, we investigated the involvement of GnIH in glycolipid metabolism and reproduction in vivo, and in the coupling between these two processes in the testis level. Our results showed that chronic intraperitoneal injection of GnIH into male mice not only increased food intake and altered meal microstructure but also significantly elevated body mass due to the increased mass of liver and epididymal white adipose tissue (eWAT), despite the loss of testicular weight. Furthermore, chronic intraperitoneal administration of GnIH to male mice resulted in obesity-related glycolipid metabolic derangements, showing hyperlipidemia, hyperglycemia, glucose intolerance, and insulin resistance through changes in the expression of glucose and lipid metabolism-related genes in the pancreas and eWAT, respectively. Interestingly, the expression of GnIH and GPR147 was markedly increased in the testis of mice under conditions of energy imbalance, such as fasting, acute hypoglycemia, and hyperglycemia. In addition, chronic GnIH injection markedly inhibited glucose and lipid metabolism of mice testis while significantly decreasing testosterone synthesis and sperm quality, inducing hypogonadism. These observations indicated that orexigenic GnIH triggers hyperphagia-induced obesity-related metabolic derangements and hypogonadism in male mice, suggesting that GnIH is an emerging candidate for coupling metabolism and fertility by involvement in obesity and metabolic disorder-induced reproductive dysfunction of the testes.
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Hiperglicemia , Hipogonadismo , Hormônios Hipotalâmicos , Animais , Glucose , Glicolipídeos , Gonadotropinas , Hiperfagia/complicações , Hipogonadismo/etiologia , Hormônios Hipotalâmicos/genética , Masculino , Camundongos , Obesidade/complicações , Sêmen/metabolismoRESUMO
RFamide-related peptide-3 (RFRP-3) has been proposed as a key inhibitory regulator of mammalian reproduction. Our previous studies demonstrated that RFRP-3 mediated apoptosis and autophagy of the epididymis in rats and inhibited porcine granulosa cell (GC) proliferation. However, the molecular mechanisms of the RFRP-3 effect on porcine GC apoptosis and autophagy have not been studied before. Herein, we first investigated the role of RFRP-3 in apoptosis and autophagy in cultured porcine GCs in vitro. Our results showed that different doses of RFRP-3 dose-dependently elevated the expression of autophagy markers at both the mRNA and protein levels, whereas the expression of apoptosis markers exhibited a bidirectional, dose-dependent effect. Because the p38MAPK signaling pathway plays essential roles in apoptosis and autophagy, we subsequently evaluated the effect of RFRP-3 on p38MAPK activation. The results showed that 10-6 M RFRP-3 treatment not only significantly decreased p38MAPK phosphorylation but also inhibited the p38MAPK activator U-46619 to promote p38MAPK activation in porcine GCs. Finally, we applied U-46619 to investigate the role of the p38MAPK signaling pathway in apoptosis and autophagy in RFRP-3-treated porcine GCs. The results showed that all doses of RFRP-3 significantly inhibited the U-46619-induced increase in apoptosis in a dose-dependent manner. However, except for the U-46619-induced Beclin-1 expression increase, which was significantly suppressed in high-dose RFRP-3-treated porcine GCs, other doses of RFRP-3 treatment strengthened the U-46619-induced increase in other autophagy markers. In summary, our data demonstrate a critical role for the p38MAPK signaling pathway in the porcine GC cellular response to RFRP-3 by controlling the balance between apoptosis and autophagy.
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Células da Granulosa , Neuropeptídeos , Proteínas Quinases p38 Ativadas por Mitógeno , Animais , Apoptose , Autofagia , Feminino , SuínosRESUMO
A highly efficient strategy for the direct thiolation of phenols under transition metal-free and solvent-free conditions has been developed. These reactions are operationally simple with employing air (molecular oxygen) as an ideal oxidant and can selectively provide mono- and dithiolation products in good to excellent yields under basic conditions. The reaction tolerates a broad range of aryl thiols and arenes and is especially applicable for large-scale synthesis.
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Understanding the drivers of forest aboveground biomass (AGB) is essential to further understanding the forest carbon cycle. In the upper Yangtze River region, where ecosystems are incredibly fragile, the driving factors that make AGB changes differ from other regions. This study aims to investigate AGB's spatial and temporal variation of Pinus densata in Shangri-La and decompose the direct and indirect effects of spatial attribute, climate, stand structure, and agricultural activity on AGB in Shangri-La to evaluate the degree of influence of each factor on AGB change. The continuous sample plots from National Forest Inventory (NFI) and Landsat time series were used to estimate the AGB in 1987, 1992, 1997, 2002, 2007, 2012, and 2017. The structural equation model (SEM) was used to analyze the different effects of the four factors on AGB based on five scales: entire, 1987-2002, 2007-2017, low population density, and high population density. The results are as follows: (1) The AGB of Pinus densata in Shangri-La decreased from 1987 to 2017, with the total amount falling from 9.52 million tons to 7.41 million tons, and the average AGB falling from 55.49 t/ha to 40.10 t/ha. (2) At different scales, stand structure and climate were the drivers that directly affect the AGB change. In contrast, the agricultural activity had a negative direct effect on the AGB change, and spatial attribute had a relatively small indirect effect on the AGB change. (3) Analyzing the SEM results at different scales, the change of the contribution of the agricultural activity indicates that human activity is the main negative driver of AGB change in Shangri-La, especially at the high population density region. In contrast, the change of the contribution of the stand structure and climate indicates that the loss of old trees has an important influence on the AGB change. Forest resources here and other ecologically fragile areas should be gradually restored by adhering to policies, such as strengthening forest protection, improving forest stand quality, and limiting agricultural production activities.