RESUMO
Recent studies on marine organisms have made use of third-generation sequencing technologies such as Pacific Biosciences (PacBio) and Oxford Nanopore Technologies (ONT). While these specialized bioinformatics tools have different algorithmic designs and performance capabilities, they offer scalability and can be applied to various datasets. We investigated the effectiveness of PacBio and ONT RNA sequencing methods in identifying the venom of the jellyfish species Nemopilema nomurai. We conducted a detailed analysis of the sequencing data from both methods, focusing on key characteristics such as CD, alternative splicing, long-chain noncoding RNA, simple sequence repeat, transcription factor, and functional transcript annotation. Our findings indicate that ONT generally produced higher raw data quality in the transcriptome analysis, while PacBio generated longer read lengths. PacBio was found to be superior in identifying CDs and long-chain noncoding RNA, whereas ONT was more cost-effective for predicting alternative splicing events, simple sequence repeats, and transcription factors. Based on these results, we conclude that PacBio is the most specific and sensitive method for identifying venom components, while ONT is the most cost-effective method for studying venogenesis, cnidocyst (venom gland) development, and transcription of virulence genes in jellyfish. Our study has implications for future sequencing technologies in marine jellyfish, and highlights the power of full-length transcriptome analysis in discovering potential therapeutic targets for jellyfish dermatitis.
Assuntos
Venenos de Cnidários , Cifozoários , Animais , RNA , Análise de Sequência de RNA , RNA não Traduzido , Sequenciamento de Nucleotídeos em Larga Escala/métodosRESUMO
Regardless of the essential role of copper (Cu) in the physiological regulation process of mammalian reproduction, excessive exposure to Cu triggers the meiotic defects of porcine oocytes via compromising the mitochondrial functions. However, the connections between the excessive Cu exposure and meiotic defects of ovine oocytes have not been reported. In this study, the effect of copper sulfate (CuSO4) exposure on the meiotic potentials of ovine oocytes was analyzed. Subsequently, the ameliorative effect of glutathione (GSH) supplementation on the meiotic defects of CuSO4 exposed ovine oocytes was investigated. For these purposes, the in vitro maturation (IVM) of ovine cumulus oocyte complexes (COCs) was conducted in the presence of 5, 10, 20 and 40 µg/mL of CuSO4 supplementation. Subsequently, different concentrations of GSH (2, 4 and 8 mM) were added to the IVM medium containing CuSO4 solution. After IVM, the assay, including nuclear maturation, spindle organization, chromosome alignment, cytoskeleton assembly, cortical granule (CGs) dynamics, mitochondrial function, reactive oxygen species (ROS) generation, apoptosis, epigenetic modification and fertilization capacity of ovine oocytes were performed. The results showed that excessive Cu exposure triggered the meiotic defects of ovine oocytes via promoting the mitochondrial dysfunction related oxidative stress damage. Moreover, the GSH supplementation, not only ameliorated the decreased maturation potential and fertilization defect of CuSO4 exposed oocytes, but inhibited the mitochondrial dysfunction related oxidative stress damage, ROS generation, apoptosis and altered H3K27me3 expression in the CuSO4 exposed oocytes. Combined with the gene expression pattern, the finding in the present study provided fundamental bases for the ameliorative effect of GSH supplementation on the meiotic defects of CuSO4 exposed oocytes via inhibiting the mitochondrial dysfunctions, further benefiting these potential applications of GSH supplementation in the mammalian IVM system and livestock breeding suffering from the excessive Cu exposure.
Assuntos
Cobre , Oócitos , Ovinos , Animais , Suínos , Cobre/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Glutationa/metabolismo , Mitocôndrias , Mamíferos/metabolismoRESUMO
Accumulating evidences revealed the connections between arsenic exposure and mitochondrial dysfunctions induced reproductive toxicology. Meanwhile, production declines were found in livestock suffering from arsenic exposure. However, the connections between arsenic exposure and livestock meiotic defects remain unclear. In this study, the effects of sodium arsenite (NaAsO2) exposure during the in vitro maturation (IVM) on the meiotic potentials of ovine oocytes were analyzed. Furthermore, the effects of glutathione (GSH) supplementation on the meiotic defects of NaAsO2 exposed ovine oocytes were investigated by the assay of nuclear maturation, spindle organization, chromosome alignment, cytoskeleton assembly, cortical granule (CGs) dynamics, mitochondrial dysfunctions, reactive oxygen species (ROS) accumulation, oxidative DNA damages, cellular apoptosis, epigenetic modifications and fertilization capacities. The results showed that the meiotic defects of NaAsO2 exposed ovine oocytes were effectively ameliorated by the GSH supplementation via the inhibition of mitochondrial dysfunctions, which not only promoted the nuclear maturation, spindle organization, chromosome alignment, cytoskeleton assembly, CGs dynamic and fertilization capacities, but also inhibited the ROS accumulation, oxidative DNA damages and apoptosis of ovine MII oocytes. The abnormal expressions of 5mC, H3K4me3 and H3K9me3 in NaAsO2 exposed ovine oocytes, indicating the abnormal epimutations of DNA methylation and histone methylation, were also effectively ameliorated by the GSH supplementation. Taken together, this study confirmed the connections between arsenic exposure and meiotic defects of ovine oocytes. Meanwhile, the effects of GSH supplementation on the developmental competence of livestock oocytes, especially for these suffering from arsenic exposure were also founded, benefiting the extended researches for the GSH applications.
RESUMO
As a popular marine saccharide, chitooligosaccharides (COS) has been proven to have good antioxidant activity. Its antioxidant effect is closely related to its degree of polymerization, degree of acetylation and sequence. However, the specific structure-activity relationship remains unclear. In this study, three chitosan dimers with different sequences were obtained by the separation and enzymatic method, and the antioxidant activity of all four chitosan dimers were studied. The effect of COS sequence on its antioxidant activity was revealed for the first time. The amino group at the reducing end plays a vital role in scavenging superoxide radicals and in the reducing power of the chitosan dimer. At the same time, we found that the fully deacetylated chitosan dimer DD showed the strongest DPPH scavenging activity. When the amino groups of the chitosan dimer were acetylated, it showed better activity in scavenging hydroxyl radicals. Research on COS sequences opens up a new path for the study of COS, and is more conducive to the investigation of its mechanism.
Assuntos
Antioxidantes/química , Quitosana/química , Sequestradores de Radicais Livres/química , Organismos Aquáticos , Compostos de Bifenilo , Humanos , Radical Hidroxila , Estrutura Molecular , PicratosRESUMO
Breast cancer (BC) is a leading cause of cancer-related death in women with unsatisfactory survival rates. Advances in the understanding of the genetic basis of BC provide the opportunity to develop gene-based medicines capable of treating metastatic diseases. Here, we first demonstrated efficient tissue engineering approaches applied to normal breast and BC extracellular matrix (ECM) starting from decellularized human biopsies to generate a three-dimensional (3D) bioactive model with the sodium lauryl ether sulfate solution. The decellularized tissues maximized the genetic component removal from tissues and minimally injured ECM structures and native compositions by histology and ECM compositions analyses. Importantly, we proved that the 3D ECM retained tissues biological properties. We demonstrated that after 30 days of recellularization with MCF-7 cell (human breast adenocarcinoma cell line), the 3D cancer ECM induced an overexpression of epithelial-mesenchymal transition (EMT) and cancer proliferation. Meanwhile, normal ECM from the breast inhibited EMT and cell growth with the inducement of apoptosis. Given the biological activity preserved in the ECM after decellularization, we believe these approaches are powerful tools for future preclinical research for BC and breast development.
Assuntos
Neoplasias da Mama/metabolismo , Matriz Extracelular/metabolismo , Alicerces Teciduais , Microambiente Tumoral , Apoptose , Neoplasias da Mama/patologia , Técnicas de Cultura de Células , Proliferação de Células , Progressão da Doença , Transição Epitelial-Mesenquimal , Feminino , Humanos , Células MCF-7 , Transdução de Sinais , Fatores de TempoRESUMO
OBJECTIVE: To study the ability of bone marrow mesenchymal stem cells (BMSCs) to repair the internal environment of the testis in male azoospermia rats. METHODS: We established azoospermia models in 22 six-week-old male SD rats by intraperitoneal injection of busulfan at 20 mg per kg body weight. We transplanted allogeneic rat BMSCs (rBMSCs) into the testicular seminiferous tubules of the model rats and, 30 days after transplantation, observed the composition and structure of the seminiferous tubular cells by HE staining and detected the expressions of CD44, CD106, and c-kit in the rBMSCs by immunohistochemistry. RESULTS: The number of epididymal sperm was significantly reduced in the model rats as compared with the normal controls (P < 0.01). CD44 and CD106, but not c-kit, were expressed in the isolated rBMSCs. At 30 days after transplantation of rBMSCs, lots of new cells were observed in the seminiferous tubules, some expressing CD106 and some expressing the germ cell surface marker c-kit. CONCLUSION: BMSCs can transdifferentiate into germ cells and repair the damaged seminiferous tubules of sterile rats.
Assuntos
Azoospermia/terapia , Transplante de Células-Tronco Mesenquimais , Túbulos Seminíferos/anatomia & histologia , Animais , Azoospermia/induzido quimicamente , Biomarcadores/metabolismo , Células da Medula Óssea , Bussulfano , Membrana Celular/metabolismo , Epididimo , Receptores de Hialuronatos/metabolismo , Masculino , Células-Tronco Mesenquimais/metabolismo , Proteínas Proto-Oncogênicas c-kit/metabolismo , Ratos , Ratos Sprague-Dawley , Túbulos Seminíferos/metabolismo , Espermatozoides , Coloração e Rotulagem , Molécula 1 de Adesão de Célula Vascular/metabolismoRESUMO
Over the past 40 years of reform and opening-up, China has achieved rapid economic and technological growth at the cost of severe air pollution. The emerging Fintech, as the result of financial institutions' adapting to the latest digital technology, might be a solution to reduce air pollution. This paper investigates the impact of Fintech development on air pollution using a two-factor fixed effects model based on data for prefecture-level cities in China from 2011 to 2017. The findings show that Fintech development can effectively reduce air pollution emissions, and this conclusion is proved to be robust throughout a series of tests. The mechanism analysis shows that Fintech reduces air pollution by promoting digital finance and green innovation.
Assuntos
Poluentes Atmosféricos , Poluição do Ar , Poluentes Atmosféricos/análise , Material Particulado/análise , Monitoramento Ambiental , Poluição do Ar/análise , Cidades , China , Desenvolvimento EconômicoRESUMO
BACKGROUND: The human body model in the virtual surgery system is generally nested by multiple complex models and each model has quite complex tangent and curvature change. In actual rendering, if all details of the human body model are rendered with high performance, it may cause the stutter due to insufficient hardware performance. If the human body model is roughly rendered, the details of the model cannot be well represented. OBJECTIVE: In order to realize the real-time rendering of complex models in virtual surgical systems, this paper proposes an improved adaptive tessellation rendering algorithm, which includes offline and online parts. METHODS: The offline part mainly completes data reading and data structure constructing. The online part performs the surface subdivision operation in-real time for each frame, which includes the subdivision operation of the control points and surface evaluation. The offline part simplifies the subdivision step by recording the surface subdivision hierarchy using a quadtree and using control templates to record control point information. RESULTS: The online part reduces computation time by using a matrix to record topological relationships between vertices and vertex weights. The online part can compress the time complexity of traversing the quadtree of different subdivision levels to Oâ¢(nâ¢logâ¡n) by establishing an association with the quadtree of each subdivision level and using the greedy algorithm to complete the traversal of the quadtree. Finally, the adaptive tessellation rendering algorithm proposed in this paper is compared with other commonly used tessellation algorithms. CONCLUSION: The algorithm has advantages in computational efficiency and graphical display.
Assuntos
Algoritmos , Interface Usuário-Computador , HumanosRESUMO
Objective: The objective of this study was to investigate the effects and mechanisms of adipose-derived stem cell-derived exosome (ADSCs-Exo) in treating premature ovarian failure (POF). Methods: We constructed a POF mouse model through intraperitoneal injection of cyclophosphamide, followed by the administration of the autophagy inhibitor 3-methyladenine (3-MA). Pathological injury, follicle stimulating hormone (FSH), malondialdehyde (MDA), reactive oxygen species (ROS), estradiol (E2), superoxide dismutase (SOD), granulosa cell (GC) apoptosis, and autophagy were assessed. Exosomes isolated from ADSCs were used to treat POF in mice. The AMPK-mTOR pathway and its proteins (p-AMPK and p-mTOR) were evaluated. A POF cell model was established using cyclophosphamide-treated human ovarian granulosa-like tumor (KGN) cells. We administered ADSCs-Exo and rapamycin to validate the mechanism of ADSCs-Exo against POF. Results: In POF mice, 3-MA treatment attenuated pathological injuries, decreased FSH, MDA, and ROS levels, and increased E2 and SOD levels. 3-MA treatment also inhibited GC apoptosis and autophagy. ADSCs-Exo alleviated pathological injuries, improved ovarian morphology and function, and reduced oxidative stress in POF mice. ADSCs-Exo inhibited GC apoptosis and autophagy. ADSCs-Exo downregulated the expression of AMPK/mTOR pathway proteins (p-AMPK and p-mTOR). In the POF cell model, ADSCs-Exo and rapamycin inhibited AMPK/mTOR-mediated autophagy. Conclusion: ADSCs-Exo inhibits POF through the inhibition of autophagy and the AMPK/mTOR pathway. This study provides a potential target for the clinical treatment of POF.
Assuntos
Exossomos , Menopausa Precoce , Insuficiência Ovariana Primária , Animais , Feminino , Humanos , Camundongos , Proteínas Quinases Ativadas por AMP/metabolismo , Autofagia , Ciclofosfamida/efeitos adversos , Exossomos/metabolismo , Hormônio Foliculoestimulante/farmacologia , Insuficiência Ovariana Primária/terapia , Espécies Reativas de Oxigênio/metabolismo , Sirolimo/farmacologia , Células-Tronco , Superóxido Dismutase/metabolismo , Serina-Treonina Quinases TOR/metabolismoRESUMO
Introduction: In the present study, the synergistic protective effect of co-supplementation of glutathione (GSH) with selenium nanoparticles (SeNPs) on the cryopreservation efficiency of bull semen was analyzed. Methods: After collection, the ejaculates of Holstein bulls were subsequently diluted with a Tris extender buffer supplemented with different concentrations of SeNPs (0, 1, 2, and 4 µg/ml), followed by semen equilibration at 4°C and assessment of sperm viability and motility. Subsequently, the ejaculates of Holstein bulls were pooled, split into four equal groups, and diluted with a Tris extender buffer supplemented with basic extender (negative control group, NC group), 2 µg/ml SeNPs (SeNPs group), 4 mM GSH (GSH group), and 4 mM GSH plus 2 µg/ml SeNPs (GSH + SeNPs group). After cryopreservation, motility, viability, mitochondrial activity, plasma membrane integrity, acrosome integrity, concentration of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT), and ability of frozen-thawed sperm cells to support in vitro embryonic development were evaluated. Results and discussion: No side effect of SeNPs concentrations applied in the current study on the motility and viability of equilibrated bull spermatozoa was found. Meanwhile, supplementation of SeNPs significantly promoted the motility and viability of equilibrated bull spermatozoa. Furthermore, the co-supplementation of GSH with SeNPs effectively protected bull spermatozoa from cryoinjury as expressed by promoting semen motility, viability, mitochondrial activity, plasma membrane integrity, and acrosome integrity. Finally, the enhanced antioxidant capacity and embryonic development potential in the frozen-thawed bull spermatozoa cryopreserved by co-supplementation of GSH with SeNPs further confirmed the synergistic protective effect of co-supplementation of GSH with SeNPs on the cryopreservation of bull semen.
RESUMO
OBJECTIVE: Stem cell therapy is a promising approach for diabetes via promoting the differentiation of insulin-producing cells (IPCs). This study aimed to screen the differentially expressed miRNAs (DEmiRNAs) during the differentiation of muscle-derived stem cells (MDSCs) into IPCs, and uncover the underlying function and mechanism of a specific DEmiRNA, miR-708-5p. METHODS: MDSCs were successfully isolated from the leg muscle of rats, and were induced for IPCs differentiation through a five-stage protocol. miRNA microarray assay was performed for screening DEmiRNAs during differentiation. The features of MDSCs-derived IPCs were identified by qRT-PCR, flow cytometry, and immunofluorescence staining. The targeting of STK4 by miR-708-5p was examined by luciferase assay. The protein expression of STK4, YAP1, and p-YAP1 was determined by Western blot and immunofluorescence staining. RESULTS: MDSCs were successfully isolated and differentiated into IPCs. A total of 12 common DEmiRNAs were obtained during five-stage differentiation. Among them, miR-708-5p that highly expressed in MDSCs-derived IPCs was selected. Overexpression of miR-708-5p upregulated some key transcription factors (Pdx1, Ngn3, Nkx2.2, Nkx6.1, Gata4, Gata6, Pax4, and Pax6) involving in IPCs differentiation, and increased insulin positive cells. In addition, STK4 was identified as the target gene of miR-708-5p. miR-708-5p overexpression downregulated the expression of STK4 and the downstream phosphorylated YAP1. CONCLUSIONS: There were 12 DEmiRNAs involved in the differentiation of MDSCs into IPCs. miR-708-5p promoted MDSCs differentiation into IPCs probably by targeting STK4-mediated Hippo-YAP1 signaling pathway.
Assuntos
Diferenciação Celular , Células Secretoras de Insulina , MicroRNAs , Mioblastos , Proteínas Serina-Treonina Quinases , Células-Tronco , Animais , Diferenciação Celular/genética , Insulina , Células Secretoras de Insulina/citologia , MicroRNAs/genética , Músculos/metabolismo , Mioblastos/citologia , Proteínas Serina-Treonina Quinases/genética , Ratos , Células-Tronco/citologiaRESUMO
Background: Ovarian cancer (OC) is the leading cause of gynecological cancer-related mortality. Verbascoside (VB) is a phenylpropanoid glycoside from Chinese herbs, with anti-tumour activities. This study aimed to investigate the effects and mechanism of VB on OC. Methods: OC cell lines SKOV3 and A2780 were used in this study. Cell viability, proliferation, and migration were measured using CCK-8, clonogenic, and transwell assays, respectively. Apoptosis and M1/M2 macrophages were detected using flow cytometry. The interaction between VB and CCN1 was predicted by molecular docking. The mRNA expression of CCN1 was detected by RT-qPCR. The protein levels of CCN1, AKT, p-AKT, p65, and p-p65 were determined by western blotting. A xenograft mice model was established for in vivo validation. Results: VB inhibited OC cell proliferation and migration in a dose-dependent manner, and promoted apoptosis and M1 macrophage polarization. VB downregulated CCN1 and inhibited the AKT/NF-κB pathway. LY294002, an AKT inhibitor, potentiated the anti-tumour effects of VB. CCN1 overexpression weakened the anti-tumour effects of VB and VB + LY294002. In vivo experiments verified that VB inhibited tumour growth and promoted M1 polarization, which is regulated by the CCN1-mediated AKT/NF-κB pathway. Conclusion: VB triggers the CCN1-AKT/NF-κB pathway-mediated M1 macrophage polarization for protecting against OC.
RESUMO
High-resolution archives deepen the understanding of past climate variability. We report new sedimentological and paleoecological data from Chaona Lake in northern China. The record represents the annually laminated (i.e., varved) archive from the western Loess Plateau spanning the Late Holocene, allowing insights into critical time intervals of decadal to centennial-scale climate instability. After developing a robust, continuously chronology supported by radioisotope dating and varve chronology, we used high-resolution palynological and sedimentological data to decipher the specific climate and ecosystem evolution over the last 2800 years. Our results show a general forest decline and climate deterioration intercalated with a series of oscillations during the Late Holocene, which may have profoundly influenced the eco-social development of northern China. In addition, lake development changes that mainly reflect the transformation from deep to shallow lake conditions generally match the regional vegetation, which is probably driven by climate-related processes. However, fluctuations in well or poor and the absence of varved sediment indicate variations in the water circulation in the lake catchment. Periods of predominantly well-varved sediments are considered to reflect reduced lake circulation and more anoxic conditions, coinciding with warmer and more stable climate intervals, such as the Roman Warm Period and the Medieval Warm Period. Conversely, periods of poor or even non-varved preservation indicate strengthened lake circulation, which may be influenced by strong winds (e.g., 2800-2000 cal yr BP) and cold/drought conditions (e.g., the Little Ice Age). Integration of our data with those of published climatic reconstructions in northern and eastern monsoon China suggests that this variability in climate can be explained by shifts in solar insolation and large-scale ocean-atmospheric coupling dynamics that affect the Loess Plateau (e.g., the East Asia Summer Monsoon and El Niño-Southern Oscillation).
Assuntos
Ecossistema , Lagos , China , Mudança Climática , El Niño Oscilação Sul , FlorestasRESUMO
Understanding the molecular level changes of oocyte cryopreservation and the subsequent warming process is essential for improving the oocyte cryopreservation technologies. Here, we collected the mature metaphase II (MII) oocytes from mice and vitrified. After thawing, single-cell whole-genome bisulphite sequencing (scWGBS) and single-cell RNA sequencing (scRNA-seq) were used to investigate the molecular attributes of this process. Compared to the fresh oocytes, the vitrified oocytes had lower global methylation and gene expression levels, and 1426 genes up-regulated and 3321 genes down-regulated. The 1426 up-regulated differentially expressed genes (DEGs) in the vitrified oocytes were mainly associated with the histone ubiquitination, while the 3321 down-regulated genes were mainly enriched in the mitochondrion organisation and ATP metabolism processes. The differentially methylated regions (DMRs) were mainly located in promoter, intron and exon region of genes, and the length of DMRs in the vitrified oocytes were also significantly lower than that of the fresh oocytes. Notably, there were no significant difference in the expression levels of DNA demethylases (Tet1, Tet2 and Tet3) and methyltransferases (Dnmt3a and Dnmt3b) between two treatments of oocytes. However, Dnmt1 and kcnq1ot1, which are responsible for maintaining DNA methylation, were significantly down regulated in the vitrified oocytes. Gene regulatory network (GRN) analysis showed the Dnmt1 and kcnq1ot1 play a core role in regulating methylation and expression levels of downstream genes. Moreover, some genes associated with oocyte quality were significantly down-regulated in the vitrified oocytes. The present data provides a new perspective for understanding the impact of vitrification on oocytes.
Assuntos
Oócitos , Vitrificação , Animais , Criopreservação/veterinária , Metilação de DNA , Expressão Gênica , Camundongos , Oócitos/metabolismo , RNA-Seq/veterináriaRESUMO
Increasing evidence indicates that cancer stem cells (CSCs) are initiators of the occurrence, development, and recurrence of malignant tumors. Mitochondria are important organelles in eukaryotic cells, not only responsible for converting part of energy released during nutrients oxidation into the energy-yielding molecule adenosine triphosphate (ATP) to fuel the activities of cell, but also play essential roles in processes such as cell apoptosis and cellular proliferation. The mitochondrial-related abnormalities have also been considered to have an important role in the origin and development of tumors. This study aimed at testing the abnormalities in mitochondrial function and energy/metabolism-related phenotypes in thyroid cancer stem cells (TCSCs). TCSCs were isolated and identified from MDA-T32 thyroid carcinoma cell line. The mitochondrial mass and mitochondrial arrangement, amount of mitochondrial DNA (mtDNA), mitochondrial membrane potential (MMP), oxygen/glucose consumption, and intracellular concentrations of reactive oxygen species (ROS) and ATP levels were examined. Perinuclear mitochondrial distribution, low amount of mtDNA and oxygen/glucose consumption, high MMP, and low intracellular ROS and ATP concentrations were observed in TCSCs. Alterations in mitochondrial function and cellular energy metabolism may be used as novel indicators of thyroid cancer.
Assuntos
Neoplasias , Glândula Tireoide , Trifosfato de Adenosina/metabolismo , DNA Mitocondrial/genética , Metabolismo Energético , Potencial da Membrana Mitocondrial , Mitocôndrias/metabolismo , Neoplasias/metabolismo , Células-Tronco Neoplásicas/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
This study investigated the effect of recombinant human lactoferrin (rhLF) on the premature ovarian failure (POF) of rats. After cyclophosphamide treatments, the POF rats were divided into the following groups: normal control group (NC), low-dose group (LD), medium-dose group (MD) and high-dose group (HD) of rhLF. After drug administrations, the ovarian indexes and hormonal levels were detected. After follicle number count, the proliferation and apoptosis were analyzed with the expressions of genes related with oogenesis, reactive oxygen species (ROS) production and apoptosis detected, followed by the calculation of oxidative stress and protein expressions. After 4-hydroperoxy cyclophosphamide (4-HC) treatments, the effect of rhLF on the proliferation, ROS production and gene expressions of primary rat granulosa cells (GCs) cultured in vitro were detected. After mating, the fertilities of POF rats were recorded. The result showed that the rhLF administrations up-regulated the ovarian index with the number of developing follicles increased and the decreases of hormonal levels conferred. The Ki-67 intensities of the MD and HD groups were up-regulated with the Tunnel intensities decreased. The rhLF treatments significantly promoted the expression of oogenesis, antioxidant and anti-apoptosis related genes. The expression of Bax and Caspase 3 were decreased with the expression of Bcl-2 up-regulated after rhLF administrations. The in vitro treatments of rhLF effectively conferred the toxicity of 4-HC on primary rat GCs. The fertility assessment showed the rhLF treatments up-regulated the offspring's' folliculogenesis, which confirmed the ameliorative role of rhLF on the POF damages via the inhibition of ROS production in GCs.
Assuntos
Anti-Infecciosos/uso terapêutico , Antineoplásicos Alquilantes/uso terapêutico , Ciclofosfamida/efeitos adversos , Lactoferrina/uso terapêutico , Insuficiência Ovariana Primária/induzido quimicamente , Animais , Anti-Infecciosos/farmacologia , Antineoplásicos Alquilantes/farmacologia , Feminino , Humanos , Lactoferrina/farmacologia , Ratos , Espécies Reativas de OxigênioRESUMO
Damage to the cell membrane is an effective method to prevent drug resistance in plant fungal diseases. Here, we proposed a negative remodeling model of the cell membrane structure induced by the C-coordinated O-carboxymethyl chitosan Cu (II) complex (O-CSLn-Cu). FITC-labeled O-CSLn-Cu (FITC-O-CSLn-Cu) was first synthesized via a nucleophilic substitution reaction and confirmed by FT-IR. FITC-labeled O-CSLn-Cu could pass through the fungal cell membrane, as detected by confocal laser scanning microscopy (CLSM) coupled with fluorescein isothiocyanate (FITC)-fluorescence. O-CSLn-Cu treatment led to apparent morphological changes in the membranes of P. capsici Leonian and giant unilamellar vesicles (GUVs) by transmission electron microscopy (TEM). Then, we performed component analysis of the cell membrane from the P. capsici Leonian affected by O-CSLn-Cu with a particular interest in membrane physicochemical properties. Many unsaturated fatty acids (UFAs) and key enzymes promoting UFA synthesis of the cell membrane were downregulated. Similarly, a large number of membrane proteins responsible for substance transport and biochemical reactions were downregulated. Furthermore, O-CSLn-Cu treatments increased plasma membrane permeability with significant leakage of intercellular electrolytes, soluble proteins and sugars, and lipid peroxidation with decreasing membrane fluidity. Finally, aquaporin 10 was proven to be a potential molecular target sensitive to antimicrobial agents according to composition analysis of membrane structure and immunohistochemistry.
Assuntos
Antifúngicos/farmacologia , Permeabilidade da Membrana Celular/efeitos dos fármacos , Quitosana/análogos & derivados , Cobre/química , Phytophthora/efeitos dos fármacos , Animais , Antifúngicos/síntese química , Antifúngicos/química , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Quitosana/química , Quitosana/farmacologia , Complexos de Coordenação/síntese química , Complexos de Coordenação/química , Complexos de Coordenação/farmacologia , Cobre/farmacologia , Fungicidas Industriais/síntese química , Fungicidas Industriais/química , Fungicidas Industriais/farmacologia , Fluidez de Membrana/efeitos dos fármacos , Lipídeos de Membrana/fisiologia , Phytophthora/metabolismo , Phytophthora/ultraestrutura , Coelhos , Esporos/efeitos dos fármacos , Esporos/fisiologiaRESUMO
Glyphosate, the active ingredient of the most widely used commercial herbicide formulation, is extensively used and produced in China. Previous studies have reported sublethal effects of glyphosate on honeybees. However, the effects of commercially formulated glyphosate (CFG) at the recommended concentration (RC) on the chronic toxicity of honeybees, especially on their behaviours, remain unknown. In this study, a series of behavioural experiments were conducted to investigate the effects of CFG on honeybees. The results showed that there was a significant decline in water responsiveness at 1/2 × , 1 × and 2 × the RC after 3 h of exposure to CFG for 11 days. The CFG significantly reduced sucrose responsiveness at 1/2 × and 1 × the RC. In addition, CFG significantly affected olfactory learning ability at 1/2 × , 1 × , and 2 × the RC and negatively affected memory ability at 1/2 × and 1 × the RC. The climbing ability of honeybees also significantly decreased at 1/2 × , 1 × and 2 × the RC. Our findings indicated that, after they were chronically exposed to CFG at the RC, honeybees exhibited behavioural changes. These results provide a theoretical basis for regulating field applications of CFG, which is necessary for establishing an early warning and notification system and for protecting honeybees.
Assuntos
Abelhas/fisiologia , Glicina/análogos & derivados , Aprendizagem/efeitos dos fármacos , Memória/efeitos dos fármacos , Atividade Motora/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Glicina/farmacologia , Herbicidas/farmacologia , GlifosatoRESUMO
With wide application in agriculture, copper fungicides have undergone three stages of development: inorganic copper, synthetic organic copper, and natural organic copper. Using chitin/chitosan (CS) as a substrate, the natural organic copper fungicide C-coordinated O-carboxymethyl chitosan Cu(II) complex (O-CSLn-Cu) was developed in the laboratory. Taking Phytophthora capsici Leonian as an example, we explored the antifungal mechanism of O-CSLn-Cu by combining tandem mass tag (TMT)-based proteomics with non-targeted liquid chromatography-mass spectrometry (LC-MS)-based metabolomics. A total of 1172 differentially expressed proteins were identified by proteomics analysis. According to the metabolomics analysis, 93 differentially metabolites were identified. Acetyl-CoA-related and membrane localized proteins showed significant differences in the proteomics analysis. Most of the differential expressed metabolites were distributed in the cytoplasm, followed by mitochondria. The integrated analysis revealed that O-CSLn-Cu could induce the "Warburg effect", with increased glycolysis in the cytoplasm and decreased metabolism in the mitochondria. Therefore, P. capsici Leonian had to compensate for ATP loss in the TCA cycle by increasing the glycolysis rate. However, this metabolic shift could not prevent the death of P. capsici Leonian. To verify this hypothesis, a series of biological experiments, such as scanning electron microscopy (SEM), transmission electron microscopy (TEM), and enzyme activity measurements were carried out. The results suggest that O-CSLn-Cu causes mitochondrial injury, which consequently leads to excessive ROS levels and insufficient ATP levels, thereby killing P. capsici Leonian.
Assuntos
Quitosana/química , Complexos de Coordenação/química , Complexos de Coordenação/farmacologia , Cobre/química , Metabolômica , Proteômica , Trifosfato de Adenosina/metabolismo , Glicólise/efeitos dos fármacos , Phytophthora/efeitos dos fármacos , Phytophthora/metabolismoRESUMO
The concept of an active dosimetry system for pulsed radiation dose rate measurements is presented. Real-time distinction of pulsed and non-pulsed radiation contributions is based on the time structure of a single interaction. A fast tissue equivalent plastic scintillator is exploited to minimize the pile-up effect influence on absorbed energy measurements. Being connected to a fully digital signal processing board, the detector creates an active dosimetry system with adjustable parameters. With this system, absorbed dose rate measurements were carried out in a photon field with a time structure mimicking a radiotherapeutic beam, but also in the presence of a constant radiation field. Measurements show a linear dependence of a pulsed radiation contribution on the accelerator current in the investigated range of the total dose rate up to 8 µGy h-1. While increasing the accelerator current by 1 µA, the pulsed radiation dose rate grows by (26.2 ± 0.9) nGy h-1 when considering pile-up events.