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Journal of Extracellular Biology Jan 2024Placental extracellular vesicles (EVs) can be found in the maternal circulation throughout gestation, and their concentration, content and bioactivity are associated...
Placental extracellular vesicles (EVs) can be found in the maternal circulation throughout gestation, and their concentration, content and bioactivity are associated with pregnancy outcomes, including gestational diabetes mellitus (GDM). However, the effect of changes in the maternal microenvironment on the mechanisms associated with the secretion of EVs from placental cells remains to be fully established. Here, we evaluated the effect of high glucose on proteins associated with the trafficking and release of different populations of EVs from placental cells. BeWo and HTR8/SVneo cells were used as placental models and cultured under 5-mM D-glucose (i.e. control) or 25-mM D-glucose (high glucose). Cell-conditioned media (CCM) and cell lysate were collected after 48 h. Different populations of EVs were isolated from CCM by ultracentrifugation (i.e. pellet 2K-g, pellet 10K-g, and pellet 100K-g) and characterised by Nanoparticle Tracking Analysis. Quantitative proteomic analysis (IDA/SWATH) and multiple reaction monitoring protocols at high resolution (MRM) were developed to quantify 37 proteins related to biogenesis, trafficking/release and recognition/uptake of EVs. High glucose increased the secretion of total EVs across the pellets from BeWo cells, an effect driven mainly by changes in the small EVs concentration in the CCM. Interestingly, no effect of high glucose on HTR8/SVneo cells EVs secretion was observed. High glucose induces changes in proteins associated with vesicle trafficking in BeWo cells, including Heat Shock Protein Family A (Hsp70) Member 9 (HSPA9) and Member 8 (HSPA8). For HTR8/SVneo, altered proteins including prostaglandin F2α receptor regulatory protein (FPRP), RAB5A, RAB35, RAB5B, and RB11B, STAM1 and TSG101. These proteins are associated with the secretion and trafficking of EVs, which could explain in part, changes in the levels of circulating EVs in diabetic pregnancies. Further, we identified that proteins RAB11B, PDCD6IP, STAM, HSPA9, HSPA8, SDCBP, RAB5B, RAB5A, RAB7A and ERAP1 regulate EV release in response to high and low glucose when overexpressed in cells. Interestingly, immunohistochemistry analysis of RAB7A revealed distinct changes in placental tissues obtained from women with normal glucose tolerance (NGT, = 6) and those with GDM ( = 6), influenced by diet or insulin treatment. High glucose regulation of proteins involved in intercellular dynamics and the trafficking of multivesicular bodies to the plasma membrane in placental cells is relevant in the context of GDM pregnancies.
PubMed: 38938672
DOI: 10.1002/jex2.135 -
PeerJ 2024Pyrophosphatases (PPases) are enzymes that catalyze the hydrolysis of pyrophosphate (PPi), a byproduct of the synthesis and degradation of diverse biomolecules. The... (Review)
Review
Pyrophosphatases (PPases) are enzymes that catalyze the hydrolysis of pyrophosphate (PPi), a byproduct of the synthesis and degradation of diverse biomolecules. The accumulation of PPi in the cell can result in cell death. Although the substrate is the same, there are variations in the catalysis and features of these enzymes. Two enzyme forms have been identified in bacteria: cytoplasmic or soluble pyrophosphatases and membrane-bound pyrophosphatases, which play major roles in cell bioenergetics. In eukaryotic cells, cytoplasmic enzymes are the predominant form of PPases (c-PPases), while membrane enzymes (m-PPases) are found only in protists and plants. The study of bacterial cytoplasmic and membrane-bound pyrophosphatases has slowed in recent years. These enzymes are central to cell metabolism and physiology since phospholipid and nucleic acid synthesis release important amounts of PPi that must be removed to allow biosynthesis to continue. In this review, two aims were pursued: first, to provide insight into the structural features of PPases known to date and that are well characterized, and to provide examples of enzymes with novel features. Second, the scientific community should continue studying these enzymes because they have many biotechnological applications. Additionally, in this review, we provide evidence that there are m-PPases present in fungi; to date, no examples have been characterized. Therefore, the diversity of PPase enzymes is still a fruitful field of research. Additionally, we focused on the roles of H/Na pumps and m-PPases in cell bioenergetics. Finally, we provide some examples of the applications of these enzymes in molecular biology and biotechnology, especially in plants. This review is valuable for professionals in the biochemistry field of protein structure-function relationships and experts in other fields, such as chemistry, nanotechnology, and plant sciences.
Topics: Inorganic Pyrophosphatase; Bacteria; Fungi; Diphosphates
PubMed: 38938619
DOI: 10.7717/peerj.17496 -
Cell Biochemistry and Function Jul 2024Multidrug resistance (MDR) during clinical chemotherapy for cancer has been considered a major obstacle to treatment efficacy. The involvement of adenosine...
Multidrug resistance (MDR) during clinical chemotherapy for cancer has been considered a major obstacle to treatment efficacy. The involvement of adenosine triphosphate-binding cassette (ABC) transporters in the MDR mechanism significantly reduces the efficacy of chemotherapeutics. This study investigates the potential of morin, a dietary bioflavonoid, to overcome colchicine resistance in KBChR-8-5 MDR cells. The P-gp inhibitory activity by morin was measured by calcein-AM drug efflux assay. Western blot analysis was employed to evaluate P-gp messenger RNA and protein expressions following morin treatment. Flow cytometry analysis and acridine orange/ethidium bromide fluorescence staining were utilised to investigate the induction of apoptosis and cell cycle arrest upon treatment with morin and paclitaxel in combination. Additionally, polymerase chain reaction (PCR) array analysis was conducted to study the gene expression profiles related to MDR, apoptosis and cell cycle arrest during treatment with morin, paclitaxel or their combination. Morin exhibited a strong binding interaction with human P-gp. This was corroborated by drug efflux assays, which showed a reduction in P-gp efflux function with increasing morin concentration. Furthermore, morin and paclitaxel combination potentiated the induction of apoptosis and G2/M phase cell cycle arrest. Morin treatment significantly downregulated the gene expression of ABCB1 and P-gp membrane expressions in MDR cells. Additionally, PCR array gene expression analysis revealed that the combination treatment with morin and paclitaxel upregulated proapoptotic and cell cycle arrest genes while downregulating ABCB1 gene and antiapoptotic genes. Thus, morin effectively reversed paclitaxel resistance in KBChR-8-5 drug-resistant cancer cells and concluded that morin resensitized the paclitaxel resistance in KBChR8-5 drug-resistant cancer cells.
Topics: Humans; Flavonoids; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Apoptosis; Paclitaxel; ATP Binding Cassette Transporter, Subfamily B, Member 1; Cell Line, Tumor; Cell Cycle Checkpoints; ATP Binding Cassette Transporter, Subfamily B; Antineoplastic Agents; Flavones
PubMed: 38938150
DOI: 10.1002/cbf.4083 -
Soft Matter Jun 2024Our ability to design artificial micro/nanomachines able to perform sophisticated tasks crucially depends on our understanding of their interaction with biosystems and...
Our ability to design artificial micro/nanomachines able to perform sophisticated tasks crucially depends on our understanding of their interaction with biosystems and their compatibility with the biological environment. Here, we design Janus colloids fuelled only by glucose and light, which can autonomously interact with cell-like compartments and trigger endocytosis. We evidence the crucial role played by the far-field hydrodynamic interaction arising from the puller/pusher swimming mode and adhesion. We show that a large contact time between the active particle and the lipid membrane is required to observe the engulfment of a particle inside a floppy giant lipid vesicle. Active Janus colloids showing relatively small velocities and a puller type swimming mode are able to target giant vesicles, deform their membranes and subsequently get stably engulfed. An instability arising from the unbound membrane segment is responsible for the transition between partial and complete stable engulfment. These experiments shed light on the physical criteria required for autonomous active particle engulfment in giant vesicles, which can serve as general principles in disciplines ranging from drug delivery and microbial infection to nanomedicine.
PubMed: 38938147
DOI: 10.1039/d4sm00337c -
ACS Nano Jun 2024Currently, specific cancer-responsive fluorogenic probes with activatable imaging and therapeutic functionalities are in great demand in the accurate diagnostics and...
Currently, specific cancer-responsive fluorogenic probes with activatable imaging and therapeutic functionalities are in great demand in the accurate diagnostics and efficient therapy of malignancies. Herein, an all-in-one strategy is presented to realize fluorescence (FL) imaging-guided and synergetic chemodynamic-photodynamic cancer therapy by using a multifunctional alkaline phosphatase (ALP)-response aggregation-induced emission (AIE) probe, TPE-APP. By responding to the abnormal expression levels of an ALP biomarker in cancer cells, the phosphate groups on the AIE probe are selectively hydrolyzed, accompanied by in situ formation of strong emissive AIE aggregates for discriminative cancer cell imaging over normal cells and highly active quinone methide species with robust chemodynamic-photodynamic activities. Consequently, the activated AIE probes can efficiently destroy cancer cell membranes and lead to the death of cancer cells within 30 min. A superior efficacy in cancer cell ablation is demonstrated in vitro and in vivo. The cancer-associated biomarker response-derived discriminative FL imaging and synergistic chemodynamic-photodynamic therapy are expected to provide a promising avenue for precise image-guided cancer therapy.
PubMed: 38938113
DOI: 10.1021/acsnano.4c03879 -
Animal Bioscience Jun 2024The Hu sheep is a renowned breed known for its high reproductive rate. It is in estrus all year round, and its breeding population is gradually expanding. However, the...
OBJECTIVE
The Hu sheep is a renowned breed known for its high reproductive rate. It is in estrus all year round, and its breeding population is gradually expanding. However, the current techniques for cryopreserving semen have limited effectiveness, which hinders the continuous development of this species. The purpose of this study is to explore the effects of different penetrating cryoprotectants (CPAs) and egg yolk (EY) concentrations on the cryopreservation of Hu ram semen to determine the most effective combination.
METHODS
In this study, the effects of glycerol (GLY), ethylene glycol (EG), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), different proportions of GLY and EG, EY on sperm quality after thawing were investigated by detecting sperm total motility (TM), progressive motility (PM), straight-line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), amplitude of lateral head displacement (ALH), wobble movement coefficient (WOB), average motion degree (MAD), functional integrity (plasma membrane integrity, acrosome integrity) and reactive oxygen species (ROS) level.
RESULTS
When GLY and EG were added together, compared to other concentration groups, 6% GLY significantly (p < 0.05) increased TM, PM, plasma membrane integrity, and acrosome integrity of thawed sperm. Additionally, it significantly (p < 0.05) decreased the ROS level of sperm. In this study, the TM, PM and membrane integrity of the 6% EG were significantly (p < 0.05) higher than those of the control, 1% GLY+5% EG and 6% GLY+6% EG groups. Compared to other concentration groups, 20% EY significantly (p < 0.05) improved the TM, PM, and plasma membrane integrity of thawed sperm. However, the integrity of the acrosome increased with the higher concentration of EY.
CONCLUSION
In conclusion, the post-thawed Hu ram semen diluted with a diluent containing 6% GLY and 20% EY exhibited higher quality compared to the other groups.
PubMed: 38938029
DOI: 10.5713/ab.24.0167 -
Advanced Science (Weinheim,... Jun 2024Recognizing the essential factor governing interfacial hydrogen/oxygen evolution reactions (HER/OER) is central to electrocatalytic water-splitting. Traditional...
Recognizing the essential factor governing interfacial hydrogen/oxygen evolution reactions (HER/OER) is central to electrocatalytic water-splitting. Traditional strategies aiming at enhancing electrocatalytic activities have mainly focused on manipulating active site valencies or coordination environments. Herein, the role of interfacial adsorption is probed and modulated by the topological construct of the electrocatalyst, a frequently underestimated non-Faradaic mechanism in the dynamics of electrocatalysis. The engineered CoFeP nanorods, anchored with FeO clusters, manifest a marked amplification of the surface electric field, thus delivering a substantially improved bifunctional electrocatalytic performance. In alkaline water splitting anion exchange membrane (AEM) electrolyzer, the current density of 1.0 A cm can be achieved at a cell voltage of only 1.73 V for the FeO@CoFeP|| FeO@CoFeP pairs for 120 h of continuous operation at 1.0 A cm. Detailed investigations of electronic structures, combined with valence state and coordination geometry assessments, reveal that the enhancement of catalytic behavior in FeO@CoFeP is chiefly attributed to the strengthened adsorptive interactions prompted by the intensified electric field at the surface. The congruent effects observed in FeO-cluster-decorated CoFeP nanosheets underscore the ubiquity of this effect. The results put forth a compelling proposition for leveraging interfacial charge densification via deliberate cluster supplementation.
PubMed: 38937998
DOI: 10.1002/advs.202403206 -
Virulence Dec 2024causes globally prevalent infections that are highly related to chronic gastritis and even development of gastric carcinomas. With the increase of antibiotic...
causes globally prevalent infections that are highly related to chronic gastritis and even development of gastric carcinomas. With the increase of antibiotic resistance, scientists have begun to search for better vaccine design strategies to eradicate colonization. However, while current strategies prefer to formulate vaccines with a single antigen, their potential has not yet been fully realized. Outer membrane vesicles (OMVs) are a potential platform since they could deliver multiple antigens. In this study, we engineered three crucial antigen proteins (UreB, CagA, and VacA) onto the surface of OMVs derived from serovar Typhimurium (. Typhimurium) mutant strains using the hemoglobin protease (Hbp) autotransporter system. In various knockout strategies, we found that OMVs isolated from the Δ Δ Δ Δ mutants could cause distinct increases in immunoglobulin G (IgG) and A (IgA) levels and effectively trigger T helper 1- and 17-biased cellular immune responses, which perform a vital role in protecting against . Next, OMVs derived from Δ Δ Δ Δ mutants were used as a vector to deliver different combinations of antigens. The antibody and cytokine levels and challenge experiments in mice model indicated that co-delivering UreB and CagA could protect against and antigen-specific T cell responses. In summary, OMVs derived from the . Typhimurium Δ Δ Δ Δ mutant strain as the vector while importing UreB and CagA as antigenic proteins using the Hbp autotransporter system would greatly benefit controlling infection.
Topics: Animals; Helicobacter Infections; Bacterial Proteins; Helicobacter pylori; Mice; Salmonella typhimurium; Antigens, Bacterial; Bacterial Vaccines; Female; Antibodies, Bacterial; Immunoglobulin G; Genetic Engineering; Urease; Disease Models, Animal
PubMed: 38937901
DOI: 10.1080/21505594.2024.2367783 -
Slit2-Robo4 signal pathway and tight junction in intestine mediate LPS-induced inflammation in mice.European Journal of Medical Research Jun 2024Sepsis is one of the most common clinical diseases, which is characterized by a serious and uncontrollable inflammatory response. LPS-induced inflammation is a critical...
BACKGROUND
Sepsis is one of the most common clinical diseases, which is characterized by a serious and uncontrollable inflammatory response. LPS-induced inflammation is a critical pathological event in sepsis, but the underlying mechanism has not yet been fully elucidated.
METHODS
The animal model was established for two batches. In the first batch of experiments, Adult C57BL/6J mice were randomly divided into control group and LPS (5 mg/kg, i.p.)group . In the second batch of experiments, mice were randomly divided into control group, LPS group, and LPS+VX765(10 mg/kg, i.p., an inhibitor of NLRP3 inflammasome) group. After 24 hours, mice were anesthetized with isoflurane, blood and intestinal tissue were collected for tissue immunohistochemistry, Western blot analysis and ELISA assays.
RESULTS
The C57BL/6J mice injected with LPS for twenty-four hours could exhibit severe inflammatory reaction including an increased IL-1β, IL-18 in serum and activation of NLRP3 inflammasome in intestine. The injection of VX765 could reverse these effects induced by LPS. These results indicated that the increased level of IL-1β and IL-18 in serum induced by LPS is related to the increased intestinal permeability and activation of NLRP3 inflammasome. In the second batch of experiments, results of western blot and immunohistochemistry showed that Slit2 and Robo4 were significant decreased in intestine of LPS group, while the expression of VEGF was significant increased. Meanwhile, the protein level of tight junction protein ZO-1, occludin, and claudin-5 were significantly lower than in control group, which could also be reversed by VX765 injection.
CONCLUSIONS
In this study, we revealed that Slit2-Robo4 signaling pathway and tight junction in intestine may be involved in LPS-induced inflammation in mice, which may account for the molecular mechanism of sepsis.
Topics: Animals; Lipopolysaccharides; Mice; Signal Transduction; Nerve Tissue Proteins; Inflammation; Intercellular Signaling Peptides and Proteins; Tight Junctions; Mice, Inbred C57BL; Male; Receptors, Cell Surface; Receptors, Immunologic; Intestinal Mucosa; NLR Family, Pyrin Domain-Containing 3 Protein; Intestines; Disease Models, Animal; Inflammasomes
PubMed: 38937814
DOI: 10.1186/s40001-024-01894-5 -
Cell & Bioscience Jun 2024Microvascular destabilization is the primary cause of the inner blood-retinal barrier (iBRB) breakdown and increased vascular leakage in diabetic retinopathy (DR).... (Review)
Review
Microvascular destabilization is the primary cause of the inner blood-retinal barrier (iBRB) breakdown and increased vascular leakage in diabetic retinopathy (DR). Microvascular destabilization results from the combinational effects of increased levels of growth factors and cytokines, involvement of inflammation, and the changed cell-to-cell interactions, especially the loss of endothelial cells and pericytes, due to hyperglycemia and hypoxia. As the manifestation of microvascular destabilization, the fluid transports via paracellular and transcellular routes increase due to the disruption of endothelial intercellular junctional complexes and/or the altered caveolar transcellular transport across the retinal vascular endothelium. With diabetes progression, the functional and the structural changes of the iBRB components, including the cellular and noncellular components, further facilitate and aggravate microvascular destabilization, resulting in macular edema, the neuroretinal damage and the dysfunction of retinal inner neurovascular unit (iNVU). Although there have been considerable recent advances towards a better understanding of the complex cellular and molecular network underlying the microvascular destabilization, some still remain to be fully elucidated. Recent data indicate that targeting the intricate signaling pathways may allow to against the microvascular destabilization. Therefore, efforts have been made to better clarify the cellular and molecular mechanisms that are involved in the microvascular destabilization in DR. In this review, we discuss: (1) the brief introduction of DR and microvascular destabilization; (2) the cellular and molecular components of iBRB and iNVU, and the breakdown of iBRB; (3) the matrix and cell-to-cell contacts to maintain microvascular stabilization, including the endothelial glycocalyx, basement membrane, and various cell-cell interactions; (4) the molecular mechanisms mediated cell-cell contacts and vascular cell death; (5) the altered cytokines and signaling pathways as well as the intricate network of the cytokines involved in microvascular destabilization. This comprehensive review aimed to provide the insights for microvascular destabilization by targeting the key molecules or specific iBRB cells, thus restoring the function and structure of iBRB and iNVU, to treat DR.
PubMed: 38937783
DOI: 10.1186/s13578-024-01269-7