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1.
Artigo em Inglês | MEDLINE | ID: mdl-35511505

RESUMO

Improving the performance of chemical transformations catalysed by microbial biocatalysts requires a deep understanding of cellular processes. While the cellular heterogeneity of cellular characteristics, such as the concentration of high abundant cellular content, is well studied, little is known about the reactivity of individual cells and its impact on the chemical identity, quantity, and purity of excreted products. Biocatalytic transformations were monitored chemically specific and quantifiable at the single-cell level by integrating droplet microfluidics, cell imaging, and mass spectrometry. Product formation rates for individual Saccharomyces cerevisiae cells were obtained by i) incubating nanolitre-sized droplets for product accumulation in microfluidic devices, ii) an imaging setup to determine the number of cells in the droplets, and iii) electrospray ionisation mass spectrometry for reading the chemical contents of individual droplets. These findings now enable the study of whole-cell biocatalysis at single-cell resolution.

2.
Front Mol Biosci ; 9: 863912, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35573729

RESUMO

In addition to the endocrine and paracrine systems, peripheral tissues such as gonads, skin, and adipose tissue are involved in the intracrine mechanisms responsible for the formation of sex steroids via the transformation of dehydroepiandrosterone and dehydroepiandrosterone sulfate (DHEA/DHEAS) into potent androgenic and estrogenic hormones. Numerous studies have examined the relationship between overweight, central obesity, and plasma levels of DHEA and DHEAS. The sodium-dependent organic anion transporter Soat (Slc10a6) is a plasma membrane uptake transporter for sulfated steroids. Significantly increased expression of Slc10a6 mRNA has been previously described in organs and tissues of lipopolysaccharide (LPS)-treated mice, including white adipose tissue. These findings suggest that Soat plays a role in the supply of steroids in peripheral target tissues. The present study aimed to investigate the expression of Soat in adipocytes and its role in adipogenesis. Soat expression was analyzed in mouse white intra-abdominal (WAT), subcutaneous (SAT), and brown (BAT) adipose tissue samples and in murine 3T3-L1 adipocytes. In addition, adipose tissue mass and size of the adipocytes were analyzed in wild-type and Slc10a6 -/- knockout mice. Soat expression was detected in mouse WAT, SAT, and BAT using immunofluorescence. The expression of Slc10a6 mRNA was significantly higher in 3T3-L1 adipocytes than that of preadipocytes and was significantly upregulated by exposure to lipopolysaccharide (LPS). Slc10a6 mRNA levels were also upregulated in the adipose tissue of LPS-treated mice. In Slc10a6 -/- knockout mice, adipocytes increased in size in the WAT and SAT of female mice and in the BAT of male mice, suggesting adipocyte hypertrophy. The serum levels of adiponectin, resistin, and leptin were comparable in wild-type and Slc10a6 -/- knockout mice. The treatment of 3T3-L1 adipocytes with DHEA significantly reduced lipid accumulation, while DHEAS did not have a significant effect. However, following LPS-induced Soat upregulation, DHEAS also significantly inhibited lipid accumulation in adipocytes. In conclusion, Soat-mediated import of DHEAS and other sulfated steroids could contribute to the complex pathways of sex steroid intracrinology in adipose tissues. Although in cell cultures the Soat-mediated uptake of DHEAS appears to reduce lipid accumulation, in Slc10a6 -/- knockout mice, the Soat deletion induced adipocyte hyperplasia through hitherto unknown mechanisms.

3.
Appl Environ Microbiol ; 88(8): e0006322, 2022 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-35369703

RESUMO

Cyanobacteria are considered promising hosts for product synthesis directly from CO2 via photosynthetic carbon assimilation. The introduction of heterologous carbon sinks in terms of product synthesis has been reported to induce the so-called "carbon sink effect," described as the release of unused photosynthetic capacity by the introduction of additional carbon. This effect is thought to arise from a limitation of carbon metabolism that represents a bottleneck in carbon and electron flow, thus enforcing a downregulation of photosynthetic efficiency. It is not known so far how the cellular source/sink balance under different growth conditions influences the extent of the carbon sink effect and in turn product formation from CO2, constituting a heterologous carbon sink. We compared the Synechocystis sp. strain PCC 6803 wild type (WT) with an engineered lactate-producing strain (SAA023) in defined metabolic states. Unexpectedly, high-light conditions combined with carbon limitation enabled additional carbon assimilation for lactate production without affecting biomass formation. Thus, a strong carbon sink effect only was observed under carbon and thus sink limitation, but not under high-sink conditions. We show that the carbon sink effect was accompanied by an increased rate of alternative electron flow (AEF). Thus, AEF plays a crucial role in the equilibration of source/sink imbalances, presumably via ATP/NADPH balancing. This study emphasizes that the evaluation of the biotechnological potential of cyanobacteria profits from cultivation approaches enabling the establishment of defined metabolic states and respective quantitative analytics. Factors stimulating photosynthesis and carbon fixation are discussed. IMPORTANCE Previous studies reported various and differing effects of the heterologous production of carbon-based molecules on photosynthetic and growth efficiency of cyanobacteria. The typically applied cultivation in batch mode, with continuously changing growth conditions, however, precludes a clear differentiation between the impact of cultivation conditions on cell physiology and effects related to the specific nature of the product and its synthesis pathway. In this study, we employed a continuous cultivation system to maintain defined source/sink conditions and thus metabolic states. This allowed a systematic and quantitative analysis of the effect of NADPH-consuming lactate production on photosynthetic and growth efficiency. This approach enables a realistic evaluation of the biotechnological potential of engineered cyanobacterial strains. For example, the quantum requirement for carbon production was found to constitute an excellent indicator of the source/sink balance and thus a key parameter for photobioprocess optimization. Such knowledge is fundamental for rational and efficient strain and process development.


Assuntos
Synechocystis , Carbono/metabolismo , Dióxido de Carbono/metabolismo , Sequestro de Carbono , Lactatos/metabolismo , NADP/metabolismo , Synechocystis/metabolismo
4.
J Clin Med ; 11(7)2022 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-35407364

RESUMO

Obesity and type 2 diabetes mellitus (T2D) represent important comorbidities of the metabolic syndrome, which are associated with non-alcoholic fatty liver disease (NAFLD)-related hepatic fibrosis. In total, 160 morbidly obese patients-81 following a low-calorie formula diet (LCD) program and 79 undergoing bariatric surgery (Roux-en-Y gastric bypass, RYGB)-were examined for anthropometric and metabolic parameters at base-line and during 12 months of weight loss, focusing on a putative co-regulation of T2D parameters and liver fibrosis risk. High NAFLD fibrosis scores (NFS) before intervention were associated with elevated HbA1c levels and T2D. Loss of weight and body fat percentage (BFL) were associated with improved glucose and lipid metabolism and reduced risk of NAFLD-related fibrosis, with particularly beneficial effects by RYGB. Both T2D improvement and NFS decrease were positively associated with high BFL. A highly significant correlation of NFS reduction with BFL was restricted to male patients while being absent in females, accompanied by generally higher BFL in men. Overall, the data display the relation of BFL, T2D improvement, and reduced NAFLD-related fibrosis risk during weight loss in morbidly obese individuals induced by diet or RYGB. Furthermore, our data suggest a considerable sexual dimorphism concerning the correlation of fat loss and improved risk of liver fibrosis.

5.
Biofilm ; 4: 100073, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35434604

RESUMO

Cyanobacteria are potent microorganisms for sustainable photo-biotechnological production processes, as they are depending mainly on water, light, and carbon dioxide. Persisting challenges preventing their application include low biomass, as well as insufficient process stability and productivity. Here, we evaluate different cyanobacteria to be applied in a novel capillary biofilm reactor. Cultivated as biofilms, the organisms self-immobilize to the reactor walls, reach high biomass and enable long and robust production processes. As 'best performer' Tolypothrix sp. PCC 7712 emerged from this study. It reached the highest biomass in the reactors with 62.6 ± 6.34 gBDWL-1, produced 0.14 µmole H2 mgChl a -1h-1 under N2-fixing conditions, showed optimal surface coverage of the available growth surface, and only minor detachment in contrast to other tested species, highlighting its potential for photobiotechnology in the near future.

6.
NPJ Breast Cancer ; 8(1): 41, 2022 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-35332139

RESUMO

The staging and local management of breast cancer involves the evaluation of the extent and completeness of excision of both the invasive carcinoma component and also the intraductal component or ductal carcinoma in situ. When both invasive ductal carcinoma and coincident ductal carcinoma in situ are present, assessment of the extent and localization of both components is required for optimal therapeutic planning. We have used a mouse model of breast cancer to evaluate the feasibility of applying molecular imaging to assess the local status of cancers in vivo. Multi-tracer positron emission tomography (PET) and magnetic resonance imaging (MRI) characterize the transition from premalignancy to invasive carcinoma. PET tracers for glucose consumption, membrane synthesis, and neoangiogenesis in combination with a Gaussian mixture model-based analysis reveal image-derived thresholds to separate the different stages within the whole-lesion. Autoradiography, histology, and quantitative image analysis of immunohistochemistry further corroborate our in vivo findings. Finally, clinical data further support our conclusions and demonstrate translational potential. In summary, this preclinical model provides a platform for characterizing multistep tumor progression and provides proof of concept that supports the utilization of advanced protocols for PET/MRI in clinical breast cancer imaging.

7.
Nat Commun ; 13(1): 1350, 2022 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-35292656

RESUMO

Magnetic skyrmions are topologically nontrivial spin textures with envisioned applications in energy-efficient magnetic information storage. Toggling the presence of magnetic skyrmions via writing/deleting processes is essential for spintronics applications, which usually require the application of a magnetic field, a gate voltage or an electric current. Here we demonstrate the reversible field-free writing/deleting of skyrmions at room temperature, via hydrogen chemisorption/desorption on the surface of Ni and Co films. Supported by Monte-Carlo simulations, the skyrmion creation/annihilation is attributed to the hydrogen-induced magnetic anisotropy change on ferromagnetic surfaces. We also demonstrate the role of hydrogen and oxygen on magnetic anisotropy and skyrmion deletion on other magnetic surfaces. Our results open up new possibilities for designing skyrmionic and magneto-ionic devices.

8.
Scand J Trauma Resusc Emerg Med ; 30(1): 23, 2022 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-35331304

RESUMO

BACKGROUND: Although prehospital point-of-care ultrasound (POCUS) is gaining in importance, its rapid interpretation remains challenging in prehospital emergency situations. The technical development of remote real-time supervision potentially offers the possibility to support emergency medicine providers during prehospital emergency ultrasound. The aim of this study was to assess the feasibility of live data transmission and supervision of prehospital POCUS in an urban environment and so to improve patients' safety. METHODS: Emergency doctors with moderate ultrasound experience performed prehospital POCUS in emergency cases (n = 24) such as trauma, acute dyspnea or cardiac shock using the portable ultrasound device Lumify™. The ultrasound examination was remotely transmitted to an emergency ultrasound expert in the clinic for real-time supervision via a secure video and audio connection. Technical feasibility as well as quality of communication and live stream were analysed. RESULTS: Prehospital POCUS with remote real-time supervision was successfully performed in 17 patients (71%). In 3 cases, the expert was not available on time and in 1 case remote data transmission was not possible due to connection problems. In 3 cases tele-supervision was restricted to video only and no verbal communication was possible via the device itself due to power saving mode of the tablet. CONCLUSION: Remote real-time supervision of prehospital POCUS in an urban environment is feasible most of the time with excellent image and communication quality. TRIAL REGISTRATION: ClinicalTrials Number NCT04612816.


Assuntos
Serviços Médicos de Emergência , Sistemas Automatizados de Assistência Junto ao Leito , Serviços Médicos de Emergência/métodos , Estudos de Viabilidade , Humanos , Testes Imediatos , Ultrassonografia/métodos
9.
Phys Med Biol ; 67(4)2022 02 16.
Artigo em Inglês | MEDLINE | ID: mdl-35171115

RESUMO

An avalanche photodiode (APD)-based small animal positron emission tomography (PET)-insert was fully evaluated for its PET performance, as well as potential influences on magnetic resonance imaging (MRI) performance. This PET-insert has an extended axial field of view (FOV) compared with the previous design to increase system sensitivity, as well as an updated cooling and temperature regulation to enable stable and reproducible PET acquisitions. The PET performance was evaluated according to the National Electrical Manufacturers Association NU4-2008 protocol. The energy and timing resolution's full width at half maximum were 16.1% and 4.7 ns, respectively. The reconstructed radial spatial resolution of the PET-insert was 1.8 mm full width at half maximum at the center FOV using filtered back projection for reconstruction and sensitivity was 3.68%. The peak noise equivalent count rates were 70 kcps for a rat-like and 350 kcps for a mouse-like phantom, respectively. Image quality phantom values and contrast recovery were comparable to state-of-the art PET-inserts and standalone systems. Regarding MR compatibility, changes in the mean signal-to-noise ratio for turbo spin echo and echo-planar imaging sequences were below 8.6%, for gradient echo sequences below 1%. Degradation of the mean homogeneity was below 2.3% for all tested sequences. The influence of the PET-insert on theB0maps was negligible and no influence on functional MRI sequences was detected. A mouse and rat imaging study demonstrated the feasibility ofin vivosimultaneous PET/MRI.


Assuntos
Avalanches , Animais , Imageamento por Ressonância Magnética/veterinária , Camundongos , Imagens de Fantasmas , Tomografia por Emissão de Pósitrons/veterinária , Ratos , Tomografia Computadorizada por Raios X
10.
Nat Mater ; 21(1): 95-102, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34663951

RESUMO

Liquids and solids are two fundamental states of matter. However, our understanding of their three-dimensional atomic structure is mostly based on physical models. Here we use atomic electron tomography to experimentally determine the three-dimensional atomic positions of monatomic amorphous solids, namely a Ta thin film and two Pd nanoparticles. We observe that pentagonal bipyramids are the most abundant atomic motifs in these amorphous materials. Instead of forming icosahedra, the majority of pentagonal bipyramids arrange into pentagonal bipyramid networks with medium-range order. Molecular dynamics simulations further reveal that pentagonal bipyramid networks are prevalent in monatomic metallic liquids, which rapidly grow in size and form more icosahedra during the quench from the liquid to the glass state. These results expand our understanding of the atomic structures of amorphous solids and will encourage future studies on amorphous-crystalline phase and glass transitions in non-crystalline materials with three-dimensional atomic resolution.

11.
Eur J Nutr ; 61(4): 1919-1929, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34951653

RESUMO

BACKGROUND: The postprandial regulation of angiopoietin-like proteins (Angptls) and their expression in adipocytes is poorly characterized. OBJECTIVE: Circulating Angptl3 and 4 were analyzed in healthy individuals undergoing either an oral lipid tolerance test (OLTT; n = 98) or an oral glucose tolerance test (OGTT; n = 99). Venous blood was drawn after 0, 2, 4, and 6 h during OLTT and after 0, 1, and 2 h during OGTT. Anthropometric and laboratory parameters were assessed and concentrations of Angptls were quantified by enzyme-linked immunosorbent assay. Angptl gene expression in 3T3-L1 adipocytes and in murine adipose tissues and cellular fractions was analyzed by quantitative real-time PCR. RESULTS: Angptl3 concentrations significantly decreased while Angptl4 levels continuously increased during OLTT. Both proteins remained unaffected during OGTT. Angptl3 and Angptl4 were expressed in murine subcutaneous and visceral AT with higher mRNA levels in mature adipocytes when compared to the stroma-vascular cell fraction. Both proteins were strongly induced during 3T3-L1 adipocyte differentiation and they were unresponsive to glucose in mature fat cells. Adipocyte Angptl3 (but not Angptl4) mRNA expression was inhibited by the polyunsaturated fatty acids arachidonic acid and docosahexaenoic acid, whereas nine types of dietary fatty acids remained without any effect. CONCLUSIONS: There is evidence of short-time regulation of Angptl3/4 levels upon metabolic stress. Angptl4 expression is high and Angptl3 expression is low in AT and restricted mainly to mature adipocytes without any differences concerning fat compartments. Whereas dietary fatty acids and glucose are without any effect, omega-3/-6-polyunsaturated fatty acids inhibited Anptl3 expression in adipocytes.

12.
Phys Rev Lett ; 127(12): 127203, 2021 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-34597075

RESUMO

It has been shown previously that the presence of a Dzyaloshinskii-Moriya interaction in perpendicularly magnetized thin films stabilizes Néel type domain walls. We demonstrate, using micromagnetic simulations and analytical modeling, that the presence of a uniaxial in plane magnetic anisotropy can also lead to the formation of Néel walls in the absence of a Dzyaloshinskii-Moriya interaction. It is possible to abruptly switch between Bloch and Néel walls via a small modulation of the in plane, but also the perpendicular, magnetic anisotropy. This opens up a route toward electric field control of the domain wall type with small applied voltages through electric field controlled anisotropies.

13.
J Clin Med ; 10(19)2021 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-34640356

RESUMO

Meteorin-like protein (Metrnl) is an adipo-myokine with pleiotropic effects in adipose tissue (AT). Its systemic regulation in obesity and under weight loss is unclear. Circulating Metrnl concentrations were analyzed by ELISA in severely obese patients undergoing bariatric surgery (BS) or low calorie diet (LCD). Metrnl mRNA expression was analyzed in human and murine tissues and cell lines by quantitative real-time PCR. About 312 morbidly obese individuals underwent BS (n = 181; BMI 53.4 + 6.8 kg/m2) or LCD (n = 131; BMI 43.5 + 6.7 kg/m2). Serum samples were obtained at baseline and 3, 6, and 12 months after intervention. AT specimen from subcutaneous and visceral adipose tissue were resected during BS. Serum Metrnl levels were lower in type 2 diabetic patients and negatively correlated with HbA1c. In BS and LCD patients, Metrnl concentrations significantly increased after 3 months and returned to baseline levels after 12 months. There was no gender-specific effect. Metrnl mRNA expression did not differ between visceral and subcutaneous AT in n = 130 patients. In contrast, Metrnl gene expression in mice was highest in intra-abdominal AT followed by subcutaneous, peri-renal, and brown AT. In the murine 3T3-L1 cell line, Metrnl expression was high in pre-adipocytes and mature adipocytes with a transient downregulation during adipocyte differentiation. Metrnl expression remained unaffected upon treatment with glucose, insulin, fatty acids, bile acids, and incretins. Polyunsaturated omega-3 and omega-6 fatty acids downregulated Metrnl expression. Systemic Metrnl is transiently upregulated during massive weight loss and gene expression in adipocytes is differentially regulated.

14.
Metab Eng ; 68: 199-209, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34673236

RESUMO

Molecular hydrogen (H2) is considered as an ideal energy carrier to replace fossil fuels in future. Biotechnological H2 production driven by oxygenic photosynthesis appears highly promising, as biocatalyst and H2 syntheses rely mainly on light, water, and CO2 and not on rare metals. This biological process requires coupling of the photosynthetic water oxidizing apparatus to a H2-producing hydrogenase. However, this strategy is impeded by the simultaneous release of oxygen (O2) which is a strong inhibitor of most hydrogenases. Here, we addressed this challenge, by the introduction of an O2-tolerant hydrogenase into phototrophic bacteria, namely the cyanobacterial model strain Synechocystis sp. PCC 6803. To this end, the gene cluster encoding the soluble, O2-tolerant, and NAD(H)-dependent hydrogenase from Ralstonia eutropha (ReSH) was functionally transferred to a Synechocystis strain featuring a knockout of the native O2 sensitive hydrogenase. Intriguingly, photosynthetically active cells produced the O2 tolerant ReSH, and activity was confirmed in vitro and in vivo. Further, ReSH enabled the constructed strain Syn_ReSH+ to utilize H2 as sole electron source to fix CO2. Syn_ReSH+ also was able to produce H2 under dark fermentative conditions as well as in presence of light, under conditions fostering intracellular NADH excess. These findings highlight a high level of interconnection between ReSH and cyanobacterial redox metabolism. This study lays a foundation for further engineering, e.g., of electron transfer to ReSH via NADPH or ferredoxin, to finally enable photosynthesis-driven H2 production.


Assuntos
Hidrogenase , Synechocystis , Hidrogênio , Hidrogenase/genética , Oxigênio , Fotossíntese , Synechocystis/genética , Synechocystis/metabolismo
15.
Int J Mol Sci ; 22(17)2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-34502345

RESUMO

Novel cultivation technologies demand the adaptation of existing analytical concepts. Metabolic flux analysis (MFA) requires stable-isotope labeling of biomass-bound protein as the primary information source. Obtaining the required protein in cultivation set-ups where biomass is inaccessible due to low cell densities and cell immobilization is difficult to date. We developed a non-disruptive analytical concept for 13C-based metabolic flux analysis based on secreted protein as an information carrier for isotope mapping in the protein-bound amino acids. This "metabolic flux probe" (MFP) concept was investigated in different cultivation set-ups with a recombinant, protein-secreting yeast strain. The obtained results grant insight into intracellular protein turnover dynamics. Experiments under metabolic but isotopically nonstationary conditions in continuous glucose-limited chemostats at high dilution rates demonstrated faster incorporation of isotope information from labeled glucose into the recombinant reporter protein than in biomass-bound protein. Our results suggest that the reporter protein was polymerized from intracellular amino acid pools with higher turnover rates than biomass-bound protein. The latter aspect might be vital for 13C-flux analyses under isotopically nonstationary conditions for analyzing fast metabolic dynamics.


Assuntos
6-Fitase/metabolismo , Isótopos de Carbono/análise , Proteínas Fúngicas/metabolismo , Glucose/metabolismo , Marcação por Isótopo/métodos , Análise do Fluxo Metabólico/métodos , Saccharomycetales/metabolismo , Isótopos de Carbono/metabolismo , Saccharomycetales/crescimento & desenvolvimento
16.
Cytokine ; 148: 155663, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34388476

RESUMO

BACKGROUND AND AIM: CAMP (Cathelicidin antimicrobial peptide) expression in adipocytes is regulated by Toll-like receptor (TLR) agonists. Secreted adipokines such as CTRP-3 have been suggested to participate in innate immune signaling in adipose tissue (AT). This study investigates whether TLR-induced CAMP expression in adipocytes is antagonized by CTRP-3. METHODS: 3T3-L1 adipocytes were co-stimulated with TLR agonists (LPS, MALP-2, Pam3CSK4, pI:C) and recombinant CTRP-3. In a SIRS model, C57BL/6 wild-type mice were intraperitoneally (ip) injected with recombinant CTRP-3 prior to LPS. CAMP expression was analyzed by real-time PCR in AT of wild-type mice and in AT and primary adipocytes from transgenic mice lacking adipocyte CTRP-3 expression. Comparative transcriptome analysis by RNA seq. was applied in CTRP-3 KO adipocytes. RESULTS: In vitro, CTRP-3 antagonized TLR4- and TLR1/2-induced CAMP expression in adipocytes whereas TLR3- and TLR2/6-mediated induction of CAMP was not affected. in vivo, application of exogenous CTRP-3 dose-dependently antagonized LPS-induced CAMP expression in intra-abdominal AT. CAMP expression in total AT and in primary adipocytes of subcutaneous and intra-abdominal AT did not differ between wild-type mice and transgenic mice lacking adipocyte CTRP-3 expression. CONCLUSIONS: The study suggests a hypothetical role of CAMP in host defense not only against Gram-positive bacteria sensed by TLR1/2 and TLR2/6 but also against Gram-negative bacteria sensed by TLR4 and potentially against viruses sensed by TLR3. The machinery of TLR-mediated pro-inflammatory activation of the CAMP gene in adipocytes seems to be partly modulated by secreted adipokines belonging to the growing family of C1q/TNF-related proteins such as CTRP-3.


Assuntos
Adipócitos/metabolismo , Adipocinas/metabolismo , Regulação da Expressão Gênica , Receptores Toll-Like/metabolismo , Células 3T3-L1 , Adipocinas/deficiência , Tecido Adiposo/metabolismo , Animais , Modelos Animais de Doenças , Lipopolissacarídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Modelos Biológicos , Síndrome de Resposta Inflamatória Sistêmica/genética , Transcriptoma/genética
17.
Cells ; 10(8)2021 08 20.
Artigo em Inglês | MEDLINE | ID: mdl-34440913

RESUMO

The C1q/TNF-related protein 3 (CTRP3) represents a pleiotropic adipokine reciprocally associated with obesity and type 2 diabetes mellitus and exhibits anti-inflammatory properties in relation to lipopolysaccharides (LPS)-mediated effects in adipocytes, as well as monocytes/macrophages. Here, we focused on the influence of CTRP3 on LPS-mediated effects in endothelial cells in order to expand the understanding of a possible anti-inflammatory function of CTRP3 in a setting of endotoxemia. An organ- and tissue-specific expression analysis by real-time PCR revealed a considerable Ctrp3 expression in various adipose tissue compartments; however, higher levels were detected in the aorta and in abundantly perfused tissues (bone marrow and the thyroid gland). We observed a robust Ctrp3 expression in primary endothelial cells and a transient upregulation in murine endothelial (MyEND) cells by LPS (50 ng/mL). In MyEND cells, CTRP3 inhibited the LPS-induced expression of interleukin (Il)-6 and the tumor necrosis factor (Tnf)-α, and suppressed the LPS-dependent expression of the major endothelial adhesion molecules Vcam-1 and Icam-1. The LPS-induced adhesion of monocytic cells to an endothelial monolayer was antagonized by CTRP3. In C57BL/6J mice with an LPS-induced systemic inflammation, exogenous CTRP3 did not affect circulating levels of TNF-α, ICAM-1, and VCAM-1. In conclusion, we characterized CTRP3 beyond its function as an adipokine in a setting of vascular inflammation. CTRP3 inhibited LPS-induced endothelial expression of adhesion molecules and monocyte cell adhesion, indicating an important vascular anti-inflammatory role for CTRP3 in endotoxemia.


Assuntos
Adipocinas/imunologia , Tecido Adiposo/imunologia , Células Endoteliais/imunologia , Perfilação da Expressão Gênica , Inflamação/imunologia , Adipocinas/genética , Adipocinas/metabolismo , Tecido Adiposo/metabolismo , Animais , Adesão Celular/efeitos dos fármacos , Adesão Celular/genética , Adesão Celular/imunologia , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/imunologia , Moléculas de Adesão Celular/metabolismo , Linhagem Celular , Citocinas/genética , Citocinas/imunologia , Citocinas/metabolismo , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Humanos , Inflamação/genética , Inflamação/metabolismo , Lipopolissacarídeos/farmacologia , Masculino , Camundongos Endogâmicos C57BL , Receptor 4 Toll-Like/genética , Receptor 4 Toll-Like/imunologia , Receptor 4 Toll-Like/metabolismo
18.
J Clin Med ; 10(15)2021 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-34362056

RESUMO

Data on the quantification of the potentially neurotrophic adipo-myokine METRNL (Meteorin-like protein) in human cerebrospinal fluid (CSF) are lacking and migration of this secreted protein across the blood-brain barrier (BBB) is uncertain. In the present pilot study, METRNL concentrations were quantified by ELISA in paired serum and CSF samples of 260 patients (107 males, 153 females) undergoing neurological evaluation. METRNL was abundant in serum (801.2 ± 378.3 pg/mL) and CSF (1007.2 ± 624.2 pg/mL) with a CSF/serum ratio of 1.4 ± 0.8. Serum METRNL levels were significantly correlated (rho = +0.521) to those in CSF. CSF METRNL concentrations were significantly correlated (rho = +0.480) with albumin CSF/serum ratios. The CSF/serum ratios of METRNL and albumin were positively correlated in Reibergram analysis (rho = 0.498), indicating that raising CSF concentrations of METRNL are mediated by increasing BBB dysfunction. The CSF concentrations of METRNL strongly increased in a stepwise manner along with increasing BBB dysfunction from grade 0 to grade 3 and with rising CSF cell count. CSF/serum ratio of METRNL also increased from grade 0 (1.2 ± 0.7) to grade 3 (3.0 ± 0.2). Furthermore, CSF levels were positively correlated with age. In conclusion, METRNL is a secreted and neurotrophic myokine that crosses over the BBB. CSF concentrations of METRNL increase with BBB dysfunction.

19.
Diagnostics (Basel) ; 11(5)2021 05 18.
Artigo em Inglês | MEDLINE | ID: mdl-34069794

RESUMO

Coronavirus disease 2019 (COVID-19) has shown the importance of postmortem investigation of deceased patients. For a correct interpretation of the pulmonary findings in this new era, it is, however, crucial to be familiar with pathologic pulmonary conditions observed in postmortem investigations in general. Adequate postmortem histopathological evaluation of the lungs may be affected by suboptimal gross work up, autolysis or poor fixation. Using a standardized preparation approach which consisted in instillation of 4% buffered formaldehyde through the large bronchi for proper fixation and preparing large frontal tissue sections of 1-2 cm thickness after at least 24 h fixation, we comprehensively analyzed postmortem pulmonary findings from consecutive adult autopsies of a two-year period before the occurrence of COVID-19 (2016-2017). In total, significant pathological findings were observed in 97/189 patients (51%), with 28 patients showing more than one pathologic condition. Acute pneumonia was diagnosed 33/128 times (26%), embolism 24 times (19%), primary pulmonary neoplasms 18 times (14%), organizing pneumonia and other fibrosing conditions 14 times (11%), pulmonary metastases 13 times (10%), diffuse alveolar damage 12 times (9%), severe emphysema 9 times (7%) and other pathologies, e.g., amyloidosis 5/128 times (4%). Pulmonary/cardiopulmonary disease was the cause of death in 60 patients (32%). Clinical and pathological diagnoses regarding lung findings correlated completely in 75 patients (40%). Autopsy led to confirmation of a clinically suspected pulmonary diagnosis in 57 patients (39%) and clarification of an unclear clinical lung finding in 16 patients (8%). Major discrepant findings regarding the lungs (N = 31; 16%) comprised cases with clinical suspicions that could not be confirmed or new findings not diagnosed intra vitam. A significant proportion of acute pneumonias (N = 8; 24% of all cases with this diagnosis; p = 0.011) was not diagnosed clinically. We confirmed the frequent occurrence of pulmonary pathologies in autopsies, including inflammatory and neoplastic lesions as the most frequent pathological findings. Acute pneumonia was an important cause for discrepancy between clinical and postmortem diagnostics.

20.
Inflammation ; 44(6): 2260-2269, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34165676

RESUMO

The anti-inflammatory adipokine CTRP-3 might affect innate immune reactions such as NOD1. The impact of CTRP-3 on NOD1-mediated inflammation in adipocytes and monocytic cells as well as on NOD1 expression was investigated. Murine 3T3-L1 pre-adipocytes and adipocytes as well as human THP-1 monocyte-like cells were co-stimulated with the synthetic NOD1 agonist Tri-DAP and recombinant CTRP-3. Gonadal adipose tissue and primary adipocytes were obtained from a murine model carrying a knockout (KO) of CTRP-3 in adipocytes but not in stroma-vascular cells. Wildtype mice with lipopolysaccharide (LPS)-induced elevated NOD1 expression were treated with CTRP-3. Secreted inflammatory cytokines in cell supernatants were measured by ELISA and mRNA levels were quantified by RT-PCR. Pro-inflammatory chemokine and cytokine secretion (MCP-1, RANTES, TNFα) was induced by NOD1 activation in adipocytes and monocyte-like cells, and MCP-1 and RANTES release was effectively inhibited by pre-incubation of cells with CTRP-3. CTRP-3 also antagonized LPS-triggered induction of NOD1 gene expression in murine adipose tissue, whereas adipocyte CTRP-3 deficiency upregulated NOD1 expression in adipose tissue. CTRP-3 is an effective antagonist of peptidoglycan-induced, NOD1-mediated inflammation and of LPS-induced NOD1 expression. Since basal NOD1 expression is increased by adipocyte CTRP-3 deficiency, there have to be also inflammation-independent mechanisms of NOD1 expression regulation by CTRP-3.


Assuntos
Adipócitos/metabolismo , Adipocinas/metabolismo , Gordura Intra-Abdominal/metabolismo , Proteína Adaptadora de Sinalização NOD1/metabolismo , Síndrome de Resposta Inflamatória Sistêmica/metabolismo , Células 3T3-L1 , Adipócitos/imunologia , Adipocinas/genética , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Humanos , Mediadores da Inflamação/metabolismo , Gordura Intra-Abdominal/imunologia , Lipopolissacarídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteína Adaptadora de Sinalização NOD1/genética , Transdução de Sinais , Gordura Subcutânea/imunologia , Gordura Subcutânea/metabolismo , Síndrome de Resposta Inflamatória Sistêmica/induzido quimicamente , Síndrome de Resposta Inflamatória Sistêmica/genética , Síndrome de Resposta Inflamatória Sistêmica/imunologia , Células THP-1
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