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1.
J Extracell Biol ; 3(4): e149, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38938848

RESUMO

Isolation of extracellular vesicles (EV) has been developing rapidly in parallel with the interest in EVs. However, commonly utilized protocols may not suit more challenging sample matrixes and could potentially yield suboptimal results. Knowing and assessing the pitfalls of isolation procedure to be used, should be involved to some extent for EV analytics. EVs in cow milk are of great interest due to their abundancy and large-scale availability as well as their cross-species bioavailability and possible use as drug carriers. However, the characteristics of milk EVs overlap with those of other milk components. This makes it difficult to isolate and study EVs individually. There exists also a lack of consensus for isolation methods. In this study, we demonstrated the differences between various differential centrifugation-based approaches for isolation of large quantities of EVs from cow milk. Samples were further purified with gradient centrifugation and size exclusion chromatography (SEC) and differences were analyzed. Quality measurements were conducted on multiple independent platforms. Particle analysis, electron microscopy and RNA analysis were used, to comprehensively characterize the isolated samples and to identify the limitations and possible sources of contamination in the EV isolation protocols. Vesicle concentration to protein ratio and RNA to protein ratios were observed to increase as samples were purified, suggesting co-isolation with major milk proteins in direct differential centrifugation protocols. We demonstrated a novel size assessment of vesicles using a particle mobility analyzer that matched the sizing using electron microscopy in contrast to commonly utilized nanoparticle tracking analysis. Based on the standards of the International Society for Extracellular Vesicles and the quick checklist of EV-Track.org for EV isolation, we emphasize the need for complete characterization and validation of the isolation protocol with all EV-related work to ensure the accuracy of results and allow further analytics and experiments.

2.
Matrix Biol ; 131: 30-45, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38788809

RESUMO

Renal development is a complex process in which two major processes, tubular branching and nephron development, regulate each other reciprocally. Our previous findings have indicated that collagen XVIII (ColXVIII), an extracellular matrix protein, affects the renal branching morphogenesis. We investigate here the role of ColXVIII in nephron formation and the behavior of nephron progenitor cells (NPCs) using isoform-specific ColXVIII knockout mice. The results show that the short ColXVIII isoform predominates in the early epithelialized nephron structures whereas the two longer isoforms are expressed only in the later phases of glomerular formation. Meanwhile, electron microscopy showed that the ColXVIII mutant embryonic kidneys have ultrastructural defects at least from embryonic day 16.5 onwards. Similar structural defects had previously been observed in adult ColXVIII-deficient mice, indicating a congenital origin. The lack of ColXVIII led to a reduced NPC population in which changes in NPC proliferation and maintenance and in macrophage influx were perceived to play a role. The changes in NPC behavior in turn led to notably reduced overall nephron formation. In conclusion, the results show that ColXVIII has multiple roles in renal development, both in ureteric branching and in NPC behavior.


Assuntos
Matriz Extracelular , Camundongos Knockout , Néfrons , Células-Tronco , Animais , Néfrons/metabolismo , Néfrons/citologia , Néfrons/crescimento & desenvolvimento , Camundongos , Matriz Extracelular/metabolismo , Células-Tronco/metabolismo , Células-Tronco/citologia , Proliferação de Células , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Colágeno/metabolismo , Colágeno/genética
3.
Plant Physiol ; 2024 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-38771246

RESUMO

Lignin is a phenolic polymer in plants that rigidifies the cell walls of water-conducting tracheary elements and support-providing fibers and stone cells. Different mechanisms have been suggested for the transport of lignin precursors to the site of lignification in the cell wall. Extracellular vesicle (EV)-enriched samples isolated from a lignin-forming cell suspension culture of Norway spruce (Picea abies L. Karst.) contained both phenolic metabolites and enzymes related to lignin biosynthesis. Metabolomic analysis revealed mono-, di- and oligolignols in the EV isolates, as well as carbohydrates and amino acids. In addition, salicylic acid (SA) and some proteins involved in SA signaling were detected in the EV-enriched samples. A proteomic analysis detected several laccases, peroxidases, ß-glucosidases, putative dirigent proteins, and cell wall-modifying enzymes such as glycosyl hydrolases, transglucosylase/hydrolases, and expansins in EVs. Our findings suggest that EVs are involved in transporting enzymes required for lignin polymerization in Norway spruce, and that radical coupling of monolignols can occur in these vesicles.

4.
Biochim Biophys Acta Mol Basis Dis ; 1870(5): 167180, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38653356

RESUMO

The renal tubular epithelial cells (TEC) have a strong capacity for repair after acute injury, but when this mechanism becomes uncontrollable, it leads to chronic kidney diseases (CKD). Indeed, in progress toward CKDs, the TECs may dedifferentiate, undergo epithelial-to-mesenchyme transition (EMT), and promote inflammation and fibrosis. Given the critical role of Wnt4 signaling in kidney ontogenesis, we addressed whether changes in this signaling are connected to renal inflammation and fibrosis by taking advantage of a knock-in Wnt4mCh/mCh mouse. While the Wnt4mCh/mCh embryos appeared normal, the corresponding mice, within one month, developed CKD-related phenotypes, such as pro-inflammatory responses including T-cell/macrophage influx, expression of fibrotic markers, and epithelial cell damage with a partial EMT. The Wnt signal transduction component ß-catenin remained unchanged, while calcium signaling is induced in the injured TECs involving Nfat and Tfeb transcription factors. We propose that the Wnt4 signaling pathway is involved in repairing the renal injury, and when the signal is overdriven, CKD is established.


Assuntos
Sinalização do Cálcio , Modelos Animais de Doenças , Transição Epitelial-Mesenquimal , Fibrose , Técnicas de Introdução de Genes , Proteína Wnt4 , Animais , Camundongos , Transição Epitelial-Mesenquimal/genética , Proteína Wnt4/metabolismo , Proteína Wnt4/genética , Sinalização do Cálcio/genética , Insuficiência Renal Crônica/patologia , Insuficiência Renal Crônica/genética , Insuficiência Renal Crônica/metabolismo , Via de Sinalização Wnt , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Rim/patologia , Rim/metabolismo , Túbulos Renais/patologia , Túbulos Renais/metabolismo , beta Catenina/metabolismo , beta Catenina/genética
5.
Behav Res Methods ; 2024 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-38389029

RESUMO

LASTU is a tool for searching for Finnish language stimulus words for psycholinguistic studies. The tool allows the user to query a number of properties, including forms, lemmas, frequencies, and morphological features. It also includes two new measures for quantifying lemma and form ambiguity. The tool is written in Python and is available for Windows and macOS platforms. It is available at https://osf.io/j8v6b/ . Included with the tool is a database based on a massive corpus of dependency-parsed Finnish language data crawled from the Internet (over 5 billion tokens). While LASTU has been developed for researchers working on the Finnish language, the openly available implementation can also be applied to other languages.

6.
Cardiovasc Res ; 120(8): 869-882, 2024 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-38289873

RESUMO

AIMS: Vascular smooth muscle cells (SMCs) and their derivatives are key contributors to the development of atherosclerosis. However, studying changes in SMC gene expression in heterogeneous vascular tissues is challenging due to the technical limitations and high cost associated with current approaches. In this paper, we apply translating ribosome affinity purification sequencing to profile SMC-specific gene expression directly from tissue. METHODS AND RESULTS: To facilitate SMC-specific translatome analysis, we generated SMCTRAP mice, a transgenic mouse line expressing enhanced green fluorescent protein (EGFP)-tagged ribosomal protein L10a (EGFP-L10a) under the control of the SMC-specific αSMA promoter. These mice were further crossed with the atherosclerosis model Ldlr-/-, ApoB100/100 to generate SMCTRAP-AS mice and used to profile atherosclerosis-associated SMCs in thoracic aorta samples of 15-month-old SMCTRAP and SMCTRAP-AS mice. Our analysis of SMCTRAP-AS mice showed that EGFP-L10a expression was localized to SMCs in various tissues, including the aortic wall and plaque. The TRAP fraction demonstrated high enrichment of known SMC-specific genes, confirming the specificity of our approach. We identified several genes, including Cemip, Lum, Mfge8, Spp1, and Serpina3, which are known to be involved in atherosclerosis-induced gene expression. Moreover, we identified several novel genes not previously linked to SMCs in atherosclerosis, such as Anxa4, Cd276, inter-alpha-trypsin inhibitor-4 (Itih4), Myof, Pcdh11x, Rab31, Serpinb6b, Slc35e4, Slc8a3, and Spink5. Among them, we confirmed the SMC-specific expression of Itih4 in atherosclerotic lesions using immunofluorescence staining of mouse aortic roots and spatial transcriptomics of human carotid arteries. Furthermore, our more detailed analysis of Itih4 showed its link to coronary artery disease through the colocalization of genome-wide association studies, splice quantitative trait loci (QTL), and protein QTL signals. CONCLUSION: We generated a SMC-specific TRAP mouse line to study atherosclerosis and identified Itih4 as a novel SMC-expressed gene in atherosclerotic plaques, warranting further investigation of its putative function in extracellular matrix stability and genetic evidence of causality.


Assuntos
Doenças da Aorta , Aterosclerose , Modelos Animais de Doenças , Músculo Liso Vascular , Miócitos de Músculo Liso , Placa Aterosclerótica , Proteínas Ribossômicas , Animais , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Aterosclerose/genética , Aterosclerose/metabolismo , Aterosclerose/patologia , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Doenças da Aorta/genética , Doenças da Aorta/patologia , Doenças da Aorta/metabolismo , Proteínas Ribossômicas/genética , Proteínas Ribossômicas/metabolismo , Camundongos Endogâmicos C57BL , Masculino , Regulação da Expressão Gênica , Perfilação da Expressão Gênica , Fenótipo , Transcriptoma , Receptores de LDL/genética , Receptores de LDL/metabolismo , Apolipoproteína B-100/genética , Apolipoproteína B-100/metabolismo , Camundongos Knockout , Humanos , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Camundongos , Camundongos Transgênicos , Aorta/metabolismo , Aorta/patologia , Feminino
7.
Nanotheranostics ; 8(1): 48-63, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38164498

RESUMO

Sweat contains biomarkers for real-time non-invasive health monitoring, but only a few relevant analytes are currently used in clinical practice. In the present study, we investigated whether sweat-derived extracellular vesicles (EVs) can be used as a source of potential protein biomarkers of human and bacterial origin. Methods: By using ExoView platform, electron microscopy, nanoparticle tracking analysis and Western blotting we characterized EVs in the sweat of eight volunteers performing rigorous exercise. We compared the presence of EV markers as well as general protein composition of total sweat, EV-enriched sweat and sweat samples collected in alginate skin patches. Results: We identified 1209 unique human proteins in EV-enriched sweat, of which approximately 20% were present in every individual sample investigated. Sweat derived EVs shared 846 human proteins (70%) with total sweat, while 368 proteins (30%) were captured by medical grade alginate skin patch and such EVs contained the typical exosome marker CD63. The majority of identified proteins are known to be carried by EVs found in other biofluids, mostly urine. Besides human proteins, EV-enriched sweat samples contained 1594 proteins of bacterial origin. Bacterial protein profiles in EV-enriched sweat were characterized by high interindividual variability, that reflected differences in total sweat composition. Alginate-based sweat patch accumulated only 5% proteins of bacterial origin. Conclusion: We showed that sweat-derived EVs provide a rich source of potential biomarkers of human and bacterial origin. Use of commercially available alginate skin patches selectively enrich for human derived material with very little microbial material collected.


Assuntos
Exossomos , Vesículas Extracelulares , Humanos , Suor/metabolismo , Vesículas Extracelulares/metabolismo , Exossomos/metabolismo , Biomarcadores/metabolismo , Alginatos/metabolismo
8.
Cell Commun Signal ; 21(1): 358, 2023 12 18.
Artigo em Inglês | MEDLINE | ID: mdl-38110951

RESUMO

BACKGROUND: During kidney organogenesis, metanephric mesenchyme (MM) and ureteric bud (UB) interact reciprocally to form nephrons. Signaling stimuli involved in these interactions include Wnts, growth factors and nano/micro particles. How UB and MM are interacting is not completely understood. Our study investigated the signaling and communication via extracellular vesicles (EVs) during nephrogenesis. Embryonic day (E) 11.5 mouse kidney UB and MM produce very low number of primary cells that have limited ability for proliferation in culture. Such limitations obstruct studying the role of EVs in induction of nephrogenesis. These issues necessitate to generate a nephrogenesis model allowing to study the comprehensive role of EVs during nephrogenesis. RESULTS: Our study generated a UB derived cell line-based in vitro flexible model of nephrogenesis allowing expandable cell culturing, in addition to performing characterization, tracking and blocking of EVs. UB cell line aggregation with E11.5 MM cells induced the formation of segmented nephrons. Most efficient nephrogenesis was obtained by the co-culturing of 30,000 cells of UB cell line with 50,000 MM cells. Results revealed that both the UB and the MM secrete EVs during nephrogenesis. UB cell line derived EVs were characterized by their size, morphology and expression of markers (CD63, TSG101, CD9 and CD81). Furthermore, proteomics data of UB cell line-derived EVs revealed large number of proteins involved in nephrogenesis-related signaling pathways. Palmitoylated GFP-tagged EVs from UB cell line were found in the nephron formation zone in the developing kidney organoid. UB cell line derived EVs did not induce nephrogenesis in MM cells but significantly contributed to the survival and nephrogenesis-competency of MM cells. The secretion of EVs was continuously inhibited during the ongoing nephrogenesis by the knockdown of RalA and RalB gene expression using short hairpin RNAs. This inhibition partially impaired the ability of UB cell line to induce nephrogenesis. Moreover, impaired nephrogenesis was partially rescued by the addition of EVs. CONCLUSION: Our study established a novel in vitro flexible model of nephrogenesis that solved the limitations of primary embryonic kidney cells and mouse embryonic stem cell kidney organoids for the EV research. EVs were found to be an integral part of nephrogenesis process. Video Abstract.


Assuntos
Vesículas Extracelulares , Rim , Animais , Camundongos , Organoides , Organogênese
9.
Front Physiol ; 14: 1295852, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38143912

RESUMO

Background: Metabolite-based sensors are attractive and highly valued for monitoring physiological parameters during rest and/or during physical activities. Owing to their molecular composition consisting of nucleic acids, proteins, and metabolites, extracellular vesicles (EVs) have become acknowledged as a novel tool for disease diagnosis. However, the evidence for sweat related EVs delivering information of physical and recovery states remains to be addressed. Methods: Taking advantage of our recently published methodology allowing the enrichment and isolation of sweat EVs from clinical patches, we investigated the metabolic load of sweat EVs in healthy participants exposed to exercise test or recovery condition. -Ten healthy volunteers (-three females and -seven males) were recruited to participate in this study. During exercise test and recovery condition, clinical patches were attached to participants' skin, on their back. Following exercise test or recovery condition, the patches were carefully removed and proceed for sweat EVs isolation. To explore the metabolic composition of sweat EVs, a targeted global metabolomics profiling of 41 metabolites was performed. Results: Our results identified seventeen metabolites in sweat EVs. These are associated with amino acids, glutamate, glutathione, fatty acids, creatine, and glycolysis pathways. Furthermore, when comparing the metabolites' levels in sweat EVs isolated during exercise to the metabolite levels in sweat EVs collected after recovery, our findings revealed a distinct metabolic profiling of sweat EVs. Furthermore, the level of these metabolites, mainly myristate, may reflect an inverse correlation with blood glucose, heart rate, and respiratory rate levels. Conclusion: Our data demonstrated that sweat EVs can be purified using routinely used clinical patches during physical activity, setting the foundations for larger-scale clinical cohort work. Furthermore, the metabolites identified in sweat EVs also offer a realistic means to identify relevant sport performance biomarkers. This study thus provides proof-of-concept towards a novel methodology that will focus on the use of sweat EVs and their metabolic composition as a non-invasive approach for developing the next-generation of sport wearable sensors.

10.
Microbiome ; 11(1): 249, 2023 11 13.
Artigo em Inglês | MEDLINE | ID: mdl-37953319

RESUMO

BACKGROUND: Reports regarding the presence of bacteria in the fetal environment remain limited and controversial. Recently, extracellular vesicles secreted by the human gut microbiota have emerged as a novel mechanism for host-microbiota interaction. We aimed to investigate the presence of bacterial extracellular vesicles in the fetal environment during healthy pregnancies and determine whether extracellular vesicles derived from the gut microbiota can cross biological barriers to reach the fetus. RESULTS: Bacterial extracellular vesicles were detectable in the amniotic fluid of healthy pregnant women, exhibiting similarities to extracellular vesicles found in the maternal gut microbiota. In pregnant mice, extracellular vesicles derived from human maternal gut microbiota were found to reach the intra-amniotic space. CONCLUSIONS: Our findings reveal maternal microbiota-derived extracellular vesicles as an interaction mechanism between the maternal microbiota and fetus, potentially playing a pivotal role in priming the prenatal immune system for gut colonization after birth. Video Abstract.


Assuntos
Vesículas Extracelulares , Microbioma Gastrointestinal , Microbiota , Gravidez , Feminino , Humanos , Camundongos , Animais , Feto/microbiologia , Líquido Amniótico/microbiologia , Bactérias
11.
Ecol Evol ; 13(10): e10612, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37841221

RESUMO

The development of DNA-based methods in recent decades has opened the door to numerous new lines of research in the biological sciences. While the speed and accuracy of DNA methodologies are clearly beneficial, the sensitivity of these methods has the adverse effect of increased susceptibility to false positives resulting from contamination in field or lab. Here, we present findings from a metabarcoding study on the diet of and food availability for five insectivorous birds, in which multiple lepidopteran species not known to occur locally were discovered. After describing the pattern of occurrences of these non-local species in the samples, we discuss various potential origins of these sequences. First, we assessed that the taxonomic assignments appeared reliable, and local occurrences of many of the species could be plausibly ruled out. Then, we looked into the possibilities of natural environmental contamination, judging it to be unlikely, albeit impossible to fully falsify. Finally, while dissimilar combinations of non-local species' occurrences across the samples did not initially suggest lab contamination, we found overlap with taxa and sequences handled in the same lab, which was undoubtedly not coincidental. Even so, not all exact sequences were accounted for in these locally conducted studies, nor was it clear if these and other sequences could remain detectable years later. Although the full explanation for the observations of non-local species remains inconclusive, these findings highlight the importance of critical examination of metabarcoding results, and showcase how species-level taxonomic assignments utilizing comprehensive reference libraries may be a tool in detecting potential contamination events, and false positives in general.

12.
Int J Mol Sci ; 24(8)2023 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-37108669

RESUMO

Cell-secreted extracellular vesicles (EVs), carrying components such as RNA, DNA, proteins, and metabolites, serve as candidates for developing non-invasive solutions for monitoring health and disease, owing to their capacity to cross various biological barriers and to become integrated into human sweat. However, the evidence for sweat-associated EVs providing clinically relevant information to use in disease diagnostics has not been reported. Developing cost-effective, easy, and reliable methodologies to investigate EVs' molecular load and composition in the sweat may help to validate their relevance in clinical diagnosis. We used clinical-grade dressing patches, with the aim being to accumulate, purify and characterize sweat EVs from healthy participants exposed to transient heat. The skin patch-based protocol described in this paper enables the enrichment of sweat EVs that express EV markers, such as CD63. A targeted metabolomics study of the sweat EVs identified 24 components. These are associated with amino acids, glutamate, glutathione, fatty acids, TCA, and glycolysis pathways. Furthermore, as a proof-of-concept, when comparing the metabolites' levels in sweat EVs isolated from healthy individuals with those of participants with Type 2 diabetes following heat exposure, our findings revealed that the metabolic patterns of sweat EVs may be linked with metabolic changes. Moreover, the concentration of these metabolites may reflect correlations with blood glucose and BMI. Together our data revealed that sweat EVs can be purified using routinely used clinical patches, setting the foundations for larger-scale clinical cohort work. Furthermore, the metabolites identified in sweat EVs also offer a realistic means to identify relevant disease biomarkers. This study thus provides a proof-of-concept towards a novel methodology that will focus on the use of the sweat EVs and their metabolites as a non-invasive approach, in order to monitor wellbeing and changes in diseases.


Assuntos
Diabetes Mellitus Tipo 2 , Vesículas Extracelulares , Humanos , Suor , Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Mellitus Tipo 2/metabolismo , Vesículas Extracelulares/metabolismo , Metabolômica , Transporte Biológico
13.
Genes (Basel) ; 14(4)2023 03 31.
Artigo em Inglês | MEDLINE | ID: mdl-37107614

RESUMO

Astrocytes are central nervous system (CNS)-restricted glial cells involved in synaptic function and CNS blood flow regulation. Astrocyte extracellular vesicles (EVs) participate in neuronal regulation. EVs carry RNAs, either surface-bound or luminal, which can be transferred to recipient cells. We characterized the secreted EVs and RNA cargo of human astrocytes derived from an adult brain. EVs were isolated by serial centrifugation and characterized with nanoparticle tracking analysis (NTA), Exoview, and immuno-transmission electron microscopy (TEM). RNA from cells, EVs, and proteinase K/RNase-treated EVs was analyzed by miRNA-seq. Human adult astrocyte EVs ranged in sizes from 50 to 200 nm, with CD81 as the main tetraspanin marker and larger EVs positive for integrin ß1. Comparison of the RNA between the cells and EVs identified RNA preferentially secreted in the EVs. In the case of miRNAs, enrichment analysis of their mRNA targets indicates that they are good candidates for mediating EV effects on recipient cells. The most abundant cellular miRNAs were also abundant in EVs, and the majority of their mRNA targets were found to be downregulated in mRNA-seq data, but the enrichment analysis lacked neuronal specificity. Proteinase K/RNase treatment of EV-enriched preparations identified RNAs secreted independently of EVs. Comparing the distribution of cellular and secreted RNA identifies the RNAs involved in intercellular communication via EVs.


Assuntos
Vesículas Extracelulares , MicroRNAs , Humanos , Adulto , Astrócitos , Endopeptidase K/genética , Transcriptoma/genética , MicroRNAs/genética , Vesículas Extracelulares/genética , RNA Mensageiro , Comunicação Celular/genética
14.
Matrix Biol ; 115: 139-159, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36623578

RESUMO

Collagen XVIII (ColXVIII) is a component of the extracellular matrix implicated in embryogenesis and control of tissue homoeostasis. We now provide evidence that ColXVIII has a specific role in renal branching morphogenesis as observed in analyses of total and isoform-specific knockout embryos and mice. The expression of the short and the two longer isoforms differ temporally and spatially during renal development. The lack of ColXVIII or its specific isoforms lead to congenital defects in the 3D patterning of the ureteric tree where the short isoform plays a prominent role. Moreover, the ex vivo data suggests that ColXVIII is involved in the kidney epithelial tree patterning via its N-terminal domains, and especially the Thrombospondin-1-like domain common to all isoforms. This morphogenetic function likely involves integrins expressed in the ureteric epithelium. Altogether, the results point to an important role for ColXVIII in the matrix-integrin-mediated functions regulating renal development.


Assuntos
Colágeno Tipo XVIII , Rim , Isoformas de Proteínas , Animais , Camundongos , Colágeno Tipo XVIII/genética , Colágeno Tipo XVIII/metabolismo , Integrinas , Rim/embriologia , Rim/metabolismo , Morfogênese , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Ureter/embriologia , Ureter/metabolismo
15.
Nat Commun ; 13(1): 6255, 2022 10 21.
Artigo em Inglês | MEDLINE | ID: mdl-36271049

RESUMO

Diabetes is a multifactorial disorder characterized by loss or dysfunction of pancreatic ß-cells. ß-cells are heterogeneous, exhibiting different glucose sensing, insulin secretion and gene expression. They communicate with other endocrine cell types via paracrine signals and between ß-cells via gap junctions. Here, we identify the importance of signaling between ß-cells via the extracellular signal WNT4. We show heterogeneity in Wnt4 expression, most strikingly in the postnatal maturation period, Wnt4-positive cells, being more mature while Wnt4-negative cells are more proliferative. Knock-out in adult ß-cells shows that WNT4 controls the activation of calcium signaling in response to a glucose challenge, as well as metabolic pathways converging to lower ATP/ADP ratios, thereby reducing insulin secretion. These results reveal that paracrine signaling between ß-cells is important in addition to gap junctions in controling insulin secretion. Together with previous reports of WNT4 up-regulation in obesity our observations suggest an adaptive insulin response coordinating ß-cells.


Assuntos
Sinalização do Cálcio , Insulinas , Glucose/metabolismo , Trifosfato de Adenosina/metabolismo , Insulinas/metabolismo , Difosfato de Adenosina/metabolismo
16.
Carbohydr Polym ; 297: 120069, 2022 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-36184157

RESUMO

The isolation of extracellular vesicles (EVs) from milk, a complex mixture of colloidal structures having a comparable size to EVs, is challenging. Although ultracentrifugation (UC) has been widely used for EV isolation, this has significant limitations, including a long processing time at high g-force conditions and large sample volume requirements. We introduced a new approach based on nature nanoentities cellulose nanofibers (CNFs) and short time and low g-force centrifugation to isolate EVs from various milk fractions. The flexible and entangled network of CNFs forms nanoporous, which entraps the EVs. Further, positively charged CNFs interact with anionic EVs through an electrostatic attraction, promoting their isolation with efficiency comparable with UC. The functionality and toxicity of isolated milk EVs were tested in Caco2 cells. Overall, the newly developed approach provides straightforward isolation and biocompatibility and preserves the natural properties of the isolated EVs, enabling further applications.


Assuntos
Vesículas Extracelulares , Nanofibras , Animais , Células CACO-2 , Celulose/farmacologia , Misturas Complexas , Humanos , Leite
17.
Development ; 149(21)2022 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-36205075

RESUMO

Kidneys develop via iterative branching of the ureteric epithelial tree and subsequent nephrogenesis at the branch points. Nephrons form in the cap mesenchyme as the metanephric mesenchyme (MM) condenses around the epithelial ureteric buds (UBs). Previous work has demonstrated that FGF8 is important for the survival of nephron progenitor cells (NPCs), and early deletion of Fgf8 leads to the cessation of nephron formation, which results in post-natal lethality. We now reveal a previously unreported function of FGF8. By combining transgenic mouse models, quantitative imaging assays and data-driven computational modelling, we show that FGF8 has a strong chemokinetic effect and that this chemokinetic effect is important for the condensation of NPCs to the UB. The computational model shows that the motility must be lower close to the UB to achieve NPC attachment. We conclude that the FGF8 signalling pathway is crucial for the coordination of NPC condensation at the UB. Chemokinetic effects have also been described for other FGFs and may be generally important for the formation of mesenchymal condensates.


Assuntos
Rim , Néfrons , Camundongos , Animais , Néfrons/metabolismo , Rim/metabolismo , Organogênese , Fatores de Crescimento de Fibroblastos/metabolismo , Células-Tronco/metabolismo , Camundongos Transgênicos , Fator 8 de Crescimento de Fibroblasto/genética , Fator 8 de Crescimento de Fibroblasto/metabolismo
18.
FASEB J ; 36(10): e22544, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36098469

RESUMO

Wnt11 regulates early cardiac development and left ventricular compaction in the heart, but it is not known how Wnt11 regulates postnatal cardiac maturation and response to cardiac stress in the adult heart. We studied cell proliferation/maturation in postnatal and adolescent Wnt11 deficient (Wnt11-/-) heart and subjected adult mice with partial (Wnt11+/-) and complete Wnt11 (Wnt11-/-) deficiency to cardiac pressure overload. In addition, we subjected primary cardiomyocytes to recombinant Wnt proteins to study their effect on cardiomyocyte growth. Wnt11 deficiency did not affect cardiomyocyte proliferation or maturation in the postnatal or adolescent heart. However, Wnt11 deficiency led to enlarged heart phenotype that was not accompanied by significant hypertrophy of individual cardiomyocytes. Analysis of stressed adult hearts from wild-type mice showed a progressive decrease in Wnt11 expression in response to pressure overload. When studied in experimental cardiac pressure overload, Wnt11 deficiency did not exacerbate cardiac hypertrophy or remodeling and cardiac function remained identical between the genotypes. When subjecting cardiomyocytes to hypertrophic stimulus, the presence of recombinant Wnt11 together with Wnt5a reduced protein synthesis. In conclusion, Wnt11 deficiency does not affect postnatal cardiomyocyte proliferation but leads to cardiac growth. Interestingly, Wnt11 deficiency alone does not substantially modulate hypertrophic response to pressure overload in vivo. Wnt11 may require cooperation with other noncanonical Wnt proteins to regulate hypertrophic response under stress.


Assuntos
Coração/crescimento & desenvolvimento , Miócitos Cardíacos/metabolismo , Proteínas Wnt/metabolismo , Animais , Cardiomegalia/metabolismo , Proliferação de Células , Camundongos , Miocárdio , Proteínas Wnt/genética
19.
Artigo em Inglês | MEDLINE | ID: mdl-35329064

RESUMO

A joint collaboration between the Arctic Centre of the University of Lapland, Finland and the Federal Institute of Industrial Research, Oshodi, Lagos, Nigeria was organised as a hybrid conference on several topics that are related to climate, food, health and entrepreneurship. The utilisation of natural resources in both regions is an important theme in meeting the sustainable development goals agenda. The topics discussed were multidisciplinary, they include Nigerian indigenous foods, bioeconomy, circular economy, nutrition, health, innovation and entrepreneurship under four themes (Climate, Food, Health and Entrepreneurship). There were dignitaries from Finland and Nigeria. The presenters are researchers from Nigerian universities (University of Ibadan, University of Abuja and Eko university, Lagos), Nigerian Federal Institute of Industrial research centre and from the Finnish side we have the university of Lapland, Rovaniemi, University of Oulu, Oulu and the Centria University of Applied Sciences, Kokkola. The topics discussed will serve as training materials for students and learners, the discussion focussed on research opportunities for institutions in both countries. The experts from both countries will continue to dialogue on the possibility of promoting common topics as research agenda in these important areas with the possibilities of creating more jobs.


Assuntos
Empreendedorismo , Desenvolvimento Sustentável , Clima , Finlândia , Humanos , Nigéria
20.
Trials ; 23(1): 118, 2022 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-35123569

RESUMO

BACKGROUND: Mindfulness-based interventions (MBIs) have been used in oncology contexts as a promising tool with numerous benefits for various health-related and psychosocial outcomes. Despite the increasing popularity of MBIs, few randomized controlled trials (RCTs) have examined their effects upon biological parameters. Specifically, no previous study has examined the effects of MBIs on extracellular vesicles (EVs), which are potentially important markers of health, disease, and stress. Moreover, the lack of RCTs is even more limited within the context of technology-mediated MBIs and long-term effects. METHODS: The current study protocol presents a two-arm, parallel, randomized controlled study investigating the effects of internet-supported mindfulness-based cognitive therapy (MBCT) compared with treatment as usual (TAU). Primary outcomes are psychological distress and EV cargo of distressed participants with previous breast, colorectal, or prostate cancer diagnoses. Secondary outcomes are self-reported psychosocial and health-related measures, and additional biological markers. Outcomes will be assessed at baseline, 4 weeks after baseline (mid-point of the intervention), 8 weeks after baseline (immediately post-intervention), 24 weeks after baseline (after booster sessions), and 52 weeks after baseline. Our goal is to recruit at least 111 participants who have been diagnosed with breast, prostate, or colorectal cancer (cancer stage I to III), are between 18 and 65 years old, and have had primary cancer treatments completed between 3 months and 5 years ago. Half of the participants will be randomized to the TAU group, and the other half will participate in an 8-week online MBCT intervention with weekly group sessions via videoconference. The intervention also includes asynchronous homework, an online retreat after the fifth week, and 4 monthly booster sessions after completion of the 8-week programme. DISCUSSION: This study will allow characterizing the effects of internet-based MBCT on psychosocial and biological indicators in the context of cancer. The effects on circulating EVs will also be investigated, as a possible neurobiological pathway underlying mind-body intervention effects. TRIAL REGISTRATION: ClinicalTrials.gov NCT04727593 (date of registration: 27 January 2021; date of record verification: 6 October 2021).


Assuntos
Terapia Cognitivo-Comportamental , Vesículas Extracelulares , Intervenção Baseada em Internet , Atenção Plena , Neoplasias , Angústia Psicológica , Adolescente , Adulto , Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias/diagnóstico , Neoplasias/terapia , Ensaios Clínicos Controlados Aleatórios como Assunto , Resultado do Tratamento , Adulto Jovem
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