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1.
Sensors (Basel) ; 24(11)2024 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-38894178

RESUMO

With the increasing demand for data exchange between nearby devices in proximity-based services, enhancing the security of wireless mutual broadcast (WMB) networks is crucial. However, WMB networks are inherently vulnerable to eavesdropping due to the open broadcast nature of their communication. This paper investigates the improvement of secrecy performance in random-access-based WMB (RA-WMB) networks by integrating physical layer security (PLS) techniques with hybrid duplex (HBD) operations under a stochastic geometry framework. The HBD method balances half-duplex (HD) receiving and full-duplex (FD) transceiving, utilizing self-interference cancellation (SIC) to enhance PLS performance. Key operational parameters, including transmission probability (TxPr), friendly jammer density, and conditions for FD operation, are designed to maximize secrecy performance. The analytical and numerical results demonstrate significant improvements in PLS performance, with SIC playing a critical role, particularly in scenarios with dense legitimate nodes, and with TxPr adjusted to balance HD receiving and FD transceiving based on SIC imperfections. The proposed design principles provide a comprehensive framework for enhancing the security of WMB networks, addressing the complex interplay of interference and SIC in various network configurations.

2.
BMC Oral Health ; 23(1): 21, 2023 01 14.
Artigo em Inglês | MEDLINE | ID: mdl-36641447

RESUMO

BACKGROUND: Delphinidin (DP), an anthocyanidin found in blueberries, has antioxidant and anti-inflammatory effects. This study aimed to investigate the efficacy of DP as a storage medium for avulsed teeth. METHODS: Human periodontal ligament cells were cultured and exposed to DP solution (10, 50, and 100 µM), Dulbecco's modified Eagle's medium, Hank's balanced salt solution and tap water. Cell counting kit-8 assays were performed after 0.5, 1, 6, and 24 h to measure the cell viability. Nitric oxide assays and gelatin zymography were performed to evaluate the anti-inflammatory effects of DP. Reverse transcription-polymerase chain reaction was used to determine the expression levels of inflammatory cytokines. RESULTS: The viability of periodontal ligament cells was greatest at 100 µM DP. At 1 h, 100 µM DP decreased nitric oxide synthesis (p < .0167). Matrix metallopeptidase-9 activity was inhibited by DP in a dose-dependent manner (p < .0167). Moreover, treatment with 100 µM DP decreased the expression levels of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-8 in periodontal ligament cells (p < .0167). CONCLUSIONS: Within the limits of this study, DP preserved the viability and suppressed the inflammatory response of periodontal ligament cells. These findings suggest that DP could be promising for preservation of avulsed teeth.


Assuntos
Soluções para Preservação de Órgãos , Avulsão Dentária , Humanos , Anti-Inflamatórios/farmacologia , Sobrevivência Celular , Óxido Nítrico , Soluções para Preservação de Órgãos/farmacologia , Ligamento Periodontal
3.
Mediators Inflamm ; 2017: 7682083, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28194047

RESUMO

We have examined the effect of progranulin (PGRN) on human T cell proliferation and its underlying mechanism. We show that PGRN inhibits the PHA-induced multiplication of T lymphocytes. It increases the number of iTregs when T lymphocytes are activated by PHA but does not do so in the absence of PHA. PGRN-mediated inhibition of T lymphocyte proliferation, as well as the induction of iTregs, was completely reversed by a TGF-ß inhibitor or a Treg inhibitor. PGRN induced TGF-ß secretion in the presence of PHA whereas it did not in the absence of PHA. Our findings indicate that PGRN suppresses T lymphocyte proliferation by enhancing the formation of iTregs from activated T lymphocytes in response to TGF-ß.


Assuntos
Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Linfócitos T/metabolismo , Linfócitos T Reguladores/metabolismo , Fator de Crescimento Transformador beta/metabolismo
4.
Bioelectromagnetics ; 36(4): 267-76, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25808160

RESUMO

This study aimed to explore effects of static magnetic fields (SMFs) of moderate intensity (3-50 mT) as biophysical stimulators of proliferation and osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells (MSCs). MSCs were exposed to SMFs of three intensities: 3, 15, and 50 mT. Proliferation was assessed by cell counting and bromodeoxyuridine incorporation, and differentiation by measuring alkaline phosphatase (ALP) activity, calcium content, mineralized nodule formation, and transcripts of osteogenic markers. Moderate intensity SMFs increased cell proliferation, ALP activity, calcium release, and mineralized nodule formation in a dose- and time-dependent manner, which peaked at 15 mT. In the same manner, they upregulated expression of osteogenic marker genes such as ALP, bone sialoprotein 2 (BSP2), collagen1a1 (COL1a1), osteocalcin (OCN), osteonectin (ON), osteopontin (OPN), osterix (OSX), and runt-related transcription factor 2 (RUNX2) with peak at 15 mT after 14 or 21 days of exposure. Results demonstrate that moderate intensity SMFs promote proliferation and osteoblastic differentiation of MSCs. This effect could help to improve MSC responses during osseointegration between a dental implant and surrounding bone.


Assuntos
Campos Magnéticos , Células-Tronco Mesenquimais/citologia , Fosfatase Alcalina/metabolismo , Calcificação Fisiológica , Cálcio/metabolismo , Proliferação de Células , Espaço Extracelular/metabolismo , Humanos , Células-Tronco Mesenquimais/metabolismo , Osteogênese
5.
Mediators Inflamm ; 2015: 794143, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26576076

RESUMO

Pathogen recognition receptors (PRRs) are a class of germ line-encoded receptors that recognize pathogen-associated molecular patterns (PAMPs). The activation of PRRs is crucial for the initiation of innate immunity, which plays a key role in first-line defense until more specific adaptive immunity is developed. PRRs differ in the signaling cascades and host responses activated by their engagement and in their tissue distribution. Currently identified PRR families are the Toll-like receptors (TLRs), the C-type lectin receptors (CLRs), the nucleotide-binding oligomerization domain-like receptors (NLRs), the retinoic acid-inducible gene-I-like receptors (RLRs), and the AIM2-like receptor (ALR). The environment of the dental pulp is substantially different from that of other tissues of the body. Dental pulp resides in a low compliance root canal system that limits the expansion of pulpal tissues during inflammatory processes. An understanding of the PRRs in dental pulp is important for immunomodulation and hence for developing therapeutic targets in the field of endodontics. Here we comprehensively review recent finding on the PRRs and the mechanisms by which innate immunity is activated. We focus on the PRRs expressed on dental pulp and periapical tissues and their role in dental pulp inflammation.


Assuntos
Polpa Dentária/imunologia , Imunidade Inata , Receptores de Reconhecimento de Padrão/fisiologia , Animais , Proteínas de Transporte/fisiologia , Polpa Dentária/microbiologia , Humanos , Lectinas Tipo C/fisiologia , Proteína 3 que Contém Domínio de Pirina da Família NLR , Proteína Adaptadora de Sinalização NOD1/fisiologia , Proteína Adaptadora de Sinalização NOD2/fisiologia , Receptores Toll-Like/fisiologia
6.
Appl Radiat Isot ; 197: 110835, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37146468

RESUMO

A simple spontaneous deposition kit for 210Po determination using alpha spectrometry was newly designed, and polonium deposition characteristics under various physicochemical conditions were evaluated using it. The high-purity silver disc (99.99%) showed high deposition efficiencies of over 85.1% in the HCl concentration range of 0.01-6 M. Optimal physicochemical factors were determined to be a temperature of 90 °C, deposition time of 90 min, and the use of ascorbic acid as a reducing agent in an amount similar to that of the interfering element (Fe).

7.
Cell Immunol ; 279(1): 1-11, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23023221

RESUMO

Mesenchymal stem cells (MSCs) have known to induce immunosuppressive properties by preventing T cell proliferation. However, it is remains unclear how MSCs inhibit T cell proliferation. To identify the factor that inhibits T cell proliferation, we conducted a cytokine array analysis of culture medium from a co-culture of MSCs and T cells and found that the chemokines, CXCL1, 2 and 3, were induced in T cells. MSCs also induced the expression of the CXCR2 receptor on T cell surface. Particularly, CXCL3 inhibited proliferation and increased apoptosis in T cells, which were reversed by CXCR2 inhibitor treatment. Moreover, CXCL3 decreased JAK2, STAT3, and AKT phosphorylation and these responses were also abolished by CXCR2 inhibitor treatment. MSCs suppressed the proliferation of T cells into tumor tissue. Collectively, these data demonstrate that MSCs directly regulate T cell proliferation by induction of CXCL3 chemokine and its receptor, CXCR2 on the surface in T cells.


Assuntos
Proliferação de Células , Células-Tronco Mesenquimais/imunologia , Receptores de Interleucina-8B/imunologia , Linfócitos T/imunologia , Animais , Animais Recém-Nascidos , Apoptose/efeitos dos fármacos , Apoptose/imunologia , Células Cultivadas , Quimiocina CXCL1/imunologia , Quimiocina CXCL1/metabolismo , Quimiocina CXCL2/imunologia , Quimiocina CXCL2/metabolismo , Quimiocinas CXC/imunologia , Quimiocinas CXC/metabolismo , Quimiocinas CXC/farmacologia , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Citometria de Fluxo , Células HeLa , Humanos , Janus Quinase 2/imunologia , Janus Quinase 2/metabolismo , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Microscopia de Fluorescência , Fosforilação , Receptores de Interleucina-8B/metabolismo , Fator de Transcrição STAT3/imunologia , Fator de Transcrição STAT3/metabolismo , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto/métodos
8.
Clin Oral Investig ; 16(4): 1153-9, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21938481

RESUMO

Bisphosphonates have been known to suppress osteoclast activity, survival, and recruitment. In this study, we tested effects of BPs on expression of two critical genes for osteoclastogenesis, M-CSF, and OPG in the process of osteoblast differentiation from hMSC. (1) The cells were cultured in osteogenic induction medium together with 0 (control group) and 10-8 M alendronate, pamidronate for up 2 and 3 weeks (for real-time PCR) and 3 and 4 weeks (for ELISA). (2) The real-time PCR protocol for M-CSF, OPG, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) consist of 40 cycles. (3) Enzyme-linked immunosorbent assay (ELISA): the amounts of M-CSF and OPG in the culture medium were determined using commercially available ELISA kits for M-CSF and OPG. Treatment of differentiating cells with alendronate or pamidronate, nitrogen-containing BPs increase the expression of OPG, which suppresses osteoclastogenesis, whereas it decreases the expression of M-CSF, which enhances preosteoclast formation. These results suggest a new mechanism by which BPs inhibit osteoclastogenesis. Results support hypothesis that progressive accumulation of bisphosphonate in jaws causes imbalance in osteogenesis and bone absorption and collateral osteoclast-osteoblast interaction. Bisphosphonate-related osteonecrosis of jaw (BPONJ) is one of the most serious complications of bisphosphonate (BP) therapy. However, the mechanism behind the this process of BPONJ is still unclear and there are so many hypotheses. Among many hypotheses, we focused on osteoclast-osteoblast interaction in this study. The findings of this study show new light on the present BPONJ occurrence theory based on the osteoclastic activity of BPs. Also, a more advanced and developed theory for BRONJ occurrence may be obtained by combining the osteoclast inhibition mechanism and the effects on osteoblastic differentiation by BPs.


Assuntos
Conservadores da Densidade Óssea/farmacologia , Difosfonatos/farmacologia , Fator Estimulador de Colônias de Macrófagos/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteoblastos/efeitos dos fármacos , Osteoprotegerina/efeitos dos fármacos , Alendronato/farmacologia , Osteonecrose da Arcada Osseodentária Associada a Difosfonatos/patologia , Técnicas de Cultura de Células , Diferenciação Celular/efeitos dos fármacos , Meios de Cultura , Ensaio de Imunoadsorção Enzimática , Gliceraldeído 3-Fosfato Desidrogenase (NADP+)/efeitos dos fármacos , Humanos , Masculino , Osteoclastos/efeitos dos fármacos , Osteogênese/fisiologia , Pamidronato , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Tempo , Adulto Jovem
9.
Talanta ; 239: 123141, 2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-34920262

RESUMO

Since daily drinking water is one of the major source for the ingestion of radiotoxic 222Rn and 226Ra, the demand for a simple method to determine these two radionuclides has significantly increased. In the present study, a rapid, simple sequential analysis method for determining 222Rn and 226Ra in drinking water using a liquid scintillation counter was developed. The method employs solvent extraction and correction equations for the effect of native 222Rn for 226Ra analysis. Validation and examination of applicability for drinking water analysis were conducted using 222Rn-injected water and 226Ra standard source. Minimum required counting times for examining drinking water on Quantulus 1220 and Hidex 300SL were estimated via minimum detectable activity depending on the counting time. In addition, the correction method, including an equation for reducing analysis time by more than 10 days, was suggested based on the analytical results for different elapsed times between sampling and measurement.


Assuntos
Água Potável , Monitoramento de Radiação , Rádio (Elemento) , Poluentes Radioativos da Água , Rádio (Elemento)/análise , Contagem de Cintilação , Poluentes Radioativos da Água/análise , Abastecimento de Água
10.
Front Cell Dev Biol ; 10: 857066, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35478967

RESUMO

Dental caries is a common disease that not only destroys the rigid structure of the teeth but also causes pulp necrosis in severe cases. Once pulp necrosis has occurred, the most common treatment is to remove the damaged pulp tissue, leading to a loss of tooth vitality and increased tooth fragility. Dental pulp stem cells (DPSCs) isolated from pulp tissue exhibit mesenchymal stem cell-like characteristics and are considered ideal candidates for regenerating damaged dental pulp tissue owing to their multipotency, high proliferation rate, and viability after cryopreservation. Importantly, DPSCs do not elicit an allogeneic immune response because they are non-immunogenic and exhibit potent immunosuppressive properties. Here, we provide an up-to-date review of the clinical applicability and potential of DPSCs, as well as emerging trends in the regeneration of damaged pulp tissue. In addition, we suggest the possibility of using DPSCs as a resource for allogeneic transplantation and provide a perspective for their clinical application in pulp regeneration.

11.
Nanoscale ; 14(13): 5138-5146, 2022 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-35302135

RESUMO

From mechanical syringes to electric field-assisted injection devices, precise control of liquid droplet generation has been sought after, and the present state-of-the-art technologies have provided droplets ranging from nanoliter to subpicoliter volume sizes. In this study, we present a new laser-driven method to generate liquid droplets with a zeptoliter volume, breaking the fundamental limits of previous studies. We guided an infrared laser beam through a hollow optical fiber (HOF) with a ring core whose end facet was coated with single-walled carbon nanotubes. The laser light was absorbed by this nanotube film and efficiently generated a highly localized microring heat source. This evaporated the liquid inside the HOF, which rapidly recondensed into zeptoliter droplets in the surrounding air at room temperature. We spectroscopically confirmed the chemical structures of the liquid precursor maintained in the droplets by atomizing dye-dissolved glycerol. Moreover, we explain the fundamental physical principles as well as functionalities of the optical atomizer and perform a detailed characterization of the droplets. Our approach has strong prospects for nanoscale delivery of biochemical substances in minuscule zeptoliter volumes.

12.
J Control Release ; 341: 646-660, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34921973

RESUMO

We report copper(II) arsenite (CuAS)-integrated polymer micelles (CuAS-PMs) as a new class of Fenton-like catalytic nanosystem that can display reactive oxygen species (ROS)-manipulating anticancer therapeutic activity. CuAS-PMs were fabricated through metal-catechol chelation-based formation of the CuAS complex on the core domain of poly (ethylene glycol)-b-poly(3,4-dihydroxy-L-phenylalanine) (PEG-PDOPA) copolymer micelles. CuAS-PMs maintained structural robustness under serum conditions. The insoluble state of the CuAS complex was effectively retained at physiological pH, whereas, at endosomal pH, the CuAS complex was ionized to release arsenite and cuprous Fenton catalysts (Cu+ ions). Upon endocytosis, CuAS-PMs simultaneously released hydrogen peroxide (H2O2)-generating arsenite and Fenton-like reaction-catalyzing Cu+ ions in cancer cells, which synergistically elevated the level of highly cytotoxic hydroxyl radicals (•OH), thereby preferentially killing cancer cells. Animal experiments demonstrated that CuAS-PMs could effectively suppress the growth of solid tumors without systemic in vivo toxicity. The design rationale of CuAS-PMs may provide a promising strategy to develop diverse oxidative stress-amplifying agents with great potential in cancer-specific therapy.


Assuntos
Antineoplásicos , Arsenitos , Nanopartículas , Animais , Antineoplásicos/química , Arsenitos/farmacologia , Cobre , Peróxido de Hidrogênio/química , Nanopartículas/química , Estresse Oxidativo
13.
Inflamm Res ; 60(5): 467-73, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21184130

RESUMO

OBJECTIVE: The aim of this study was to investigate effects of CD137 ligand (CD137L)-mediated reverse signaling on cellular responses in thioglycollate-elicited mouse peritoneal macrophages. METHODS: Five-week-old male C57B6 mice were injected intraperitoneally with thioglycollate for 3 days to isolate peritoneal macrophages. We counted total cell numbers with a Cell Lab Quanta SC. CD137L expression was examined with a flow cytometer. We also measured expression of inflammatory cytokines by flow cytometry and/or real time-PCR. RESULTS: Cross-linking of CD137L with recombinant CD137-Fc protein (rCD137-Fc) increased total numbers of thioglycollate-elicited mouse peritoneal macrophages (hIgG-Fc- vs. rCD137-Fc-treated group, p < 0.05). However, ligation reduced the increase in IL-1ß and IL-6 levels. Real-time PCR analysis showed that treatment of cells with rCD137-Fc also reduced transcript levels of IL-1ß, IL-6, iNOS and COX2 (hIgG-Fc- vs. rCD137-Fc-treated group, p < 0.05), as well as expression of CD137L. CONCLUSION: Reverse signals initiated by CD137L negatively modulate certain immune functions of thioglycollate-elicited peritoneal macrophages.


Assuntos
Ligante 4-1BB/química , Reagentes de Ligações Cruzadas/farmacologia , Macrófagos Peritoneais/metabolismo , Tioglicolatos/química , Ligante 4-1BB/metabolismo , Animais , Regulação para Baixo , Citometria de Fluxo/métodos , Sistema Imunitário , Interleucina-1beta/biossíntese , Interleucina-6/biossíntese , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Transdução de Sinais
14.
J Nanosci Nanotechnol ; 11(7): 6371-6, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22121718

RESUMO

Electrospun Nanofiber sheets have been shown to mimic the structure of extracellular matrix (ECM). Although these nanofibers have shown great potential for use as tissue engineering scaffolds, it is difficult for the electrospun nanofiber based sheets to be shaped into the desired three-dimensional structure. In this study, poly(L-lactic acid) (PLLA), a biodegradable and biocompatible polyester, was electrospun to produce nanofibers that were treated with an amino group containing base in order to fabricate polymeric nanocylinders. The aspect ratio of the PLLA nanocylinders was tunable by varying the aminolysis time and density of the amino group containing base. The effects of changes in nanofibrous morphology of the PLLA nanocylinders/macro-porous gelatin scaffolds on cell adhesion and proliferation were evaluated. The results revealed different cell morphology, adhesion, and proliferation in the nanocylinders composite gelatin scaffold versus gelatin scaffold alone. Confocal laser scanning microscopy observation showed more spreading and a more flattened cell morphology after NIH3T3 cells were cultured on PLLA nanocylinders/gelatin scaffolds for 10 hours and 4 days. These results indicate that the gelatin/PLLA nanocylinder composite is a promising way to fabricate 3D nanofibrous scaffolds that accelerates cell adhesion and proliferation for tissue engineering.


Assuntos
Gelatina/química , Ácido Láctico/química , Nanotubos/química , Polímeros/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ácido Láctico/farmacologia , Teste de Materiais , Camundongos , Microscopia Confocal , Microscopia Eletrônica de Varredura , Células NIH 3T3 , Nanofibras , Nanotecnologia , Poliésteres , Polímeros/farmacologia , Porosidade , Espectroscopia de Infravermelho com Transformada de Fourier
15.
Sci Rep ; 11(1): 12690, 2021 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-34135451

RESUMO

We report a new method to optically manipulate a single dielectric particle along closed-loop polygonal trajectories by crossing a suite of all-fiber Bessel-like beams within a single water droplet. Exploiting optical radiation pressure, this method demonstrates the circulation of a single polystyrene bead in both a triangular and a rectangle geometry enabling the trapped particle to undergo multiple circulations successfully. The crossing of the Bessel-like beams creates polygonal corners where the trapped particles successfully make abrupt turns with acute angles, which is a novel capability in microfluidics. This offers an optofluidic paradigm for particle transport overcoming turbulences in conventional microfluidic chips.

16.
Cell Mol Immunol ; 18(8): 1956-1968, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-32868911

RESUMO

The antitumor capabilities of agonistic anti-4-1BB mAbs have made them an attractive target for tumor immunotherapy. However, the adverse side effects associated with agonist antibodies have hindered their clinical development. Here, we aimed to study the immune-related adverse events of repeated doses and long-term use of agonistic anti-4-1BB mAbs. We show that chronic activation of 4-1BB signals induced the accumulation of IFN-γ-producing PD-1+CD8+ T cells in the secondary lymphoid organs of tumor-bearing mice by increasing the number of dividing CD8+ T cells, which was beneficial for suppressing tumor growth in the early phase of anti-4-1BB induction. However, repeated exposure to anti-4-1BB mAbs led to granuloma development in tumor-draining lymph nodes (TDLNs) of mice due to recruitment and accumulation of macrophages via the CD8+ T cell-IFN-γ axis. This was accompanied by excessive lymph node swelling, which impaired the sequential activation of CD8+ T cells. Our data provide insights into the immune-related adverse events of long-term agonist 4-1BB antibody dosing, which should be considered during the clinical development of immunomodulating therapy.


Assuntos
Linfócitos T CD8-Positivos , Neoplasias , Animais , Granuloma/patologia , Linfonodos , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias/patologia , Membro 9 da Superfamília de Receptores de Fatores de Necrose Tumoral
17.
Cell Syst ; 12(6): 670-687.e10, 2021 06 16.
Artigo em Inglês | MEDLINE | ID: mdl-34043964

RESUMO

Although some cell types may be defined anatomically or by physiological function, a rigorous definition of cell state remains elusive. Here, we develop a quantitative, imaging-based platform for the systematic and automated classification of subcellular organization in single cells. We use this platform to quantify subcellular organization and gene expression in >30,000 individual human induced pluripotent stem cell-derived cardiomyocytes, producing a publicly available dataset that describes the population distributions of local and global sarcomere organization, mRNA abundance, and correlations between these traits. While the mRNA abundance of some phenotypically important genes correlates with subcellular organization (e.g., the beta-myosin heavy chain, MYH7), these two cellular metrics are heterogeneous and often uncorrelated, which suggests that gene expression alone is not sufficient to classify cell states. Instead, we posit that cell state should be defined by observing full distributions of quantitative, multidimensional traits in single cells that also account for space, time, and function.


Assuntos
Células-Tronco Pluripotentes Induzidas , Diferenciação Celular/genética , Humanos , Miócitos Cardíacos/metabolismo , Transcriptoma/genética
18.
Eur J Immunol ; 39(9): 2617-28, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19676073

RESUMO

Cross-linking of CD137 ligand (CD137L), a member of the TNF family, with recombinant CD137-Fc (rCD137-Fc) protein enhanced adherence of bone marrow-derived macrophages, and increased the expression of ICAM-1, IL-1beta, IL-6, M-CSF and phosphotyrosine proteins. In RAW264.7 cells, a murine myeloid cell line, rCD137-Fc not only increased adherence but also cell multiplication, in a manner comparable to LPS or M-CSF. In addition, it up-regulated expression of IL-1beta, IL-1 receptor antagonist, IL-6, COX2, tenascin C, neuropeptide Y and M-CSF mRNA. Neutralization of M-CSF by incubating the RAW264.7 cells with anti-M-CSF mAb did not prevent the CD137L signal-induced viability. Viability was blocked by PP2, an Src tyrosine kinase inhibitor, rapamycin, an mTOR inhibitor and LY294002, a PI3K inhibitor, but not by Wortmannin, another PI3K inhibitor. Cross-linking of CD137L increased phosphorylation of Akt and p70S6 kinase. The latter was blocked by PP2, rapamycin or LY294002, but not by Wortmannin, whereas phosphorylation of Akt was blocked by LY294002 or Wortmannin. These findings demonstrate that reverse signals evoked by CD137L regulate immune functions in macrophages.


Assuntos
Ligante 4-1BB/metabolismo , Proteínas de Transporte/imunologia , Sobrevivência Celular/imunologia , Macrófagos/imunologia , Fosfotransferases (Aceptor do Grupo Álcool)/imunologia , Proteínas Proto-Oncogênicas c-akt/imunologia , Ligante 4-1BB/imunologia , Animais , Proteínas de Transporte/antagonistas & inibidores , Proteínas de Transporte/metabolismo , Adesão Celular/efeitos dos fármacos , Adesão Celular/imunologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cromonas/farmacologia , Citocinas/efeitos dos fármacos , Citocinas/imunologia , Citocinas/metabolismo , Inibidores Enzimáticos/farmacologia , Imunidade Inata , Imunossupressores/farmacologia , Molécula 1 de Adesão Intercelular/efeitos dos fármacos , Molécula 1 de Adesão Intercelular/imunologia , Molécula 1 de Adesão Intercelular/metabolismo , Interleucina-1beta/agonistas , Interleucina-1beta/imunologia , Interleucina-1beta/metabolismo , Interleucina-6/agonistas , Interleucina-6/imunologia , Interleucina-6/metabolismo , Lipopolissacarídeos/farmacologia , Fator Estimulador de Colônias de Macrófagos/agonistas , Fator Estimulador de Colônias de Macrófagos/imunologia , Fator Estimulador de Colônias de Macrófagos/metabolismo , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Morfolinas/farmacologia , Células Mieloides/efeitos dos fármacos , Células Mieloides/imunologia , Fosforilação/efeitos dos fármacos , Fosforilação/imunologia , Fosfotransferases (Aceptor do Grupo Álcool)/antagonistas & inibidores , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Pirimidinas/farmacologia , Proteínas Recombinantes/farmacologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Sirolimo/farmacologia , Serina-Treonina Quinases TOR , Membro 9 da Superfamília de Receptores de Fatores de Necrose Tumoral/farmacologia , Quinases da Família src/antagonistas & inibidores , Quinases da Família src/imunologia , Quinases da Família src/metabolismo
19.
Cryobiology ; 60(3): 326-30, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20214895

RESUMO

Gene expression was compared by cDNA microarray analysis in human periodontal ligament (PDL) cells cultured from teeth immediately after extraction and from teeth cryopreserved for 1 week. Twenty healthy collateral premolar teeth without caries and restorations were obtained from 10 young patients, one maxillary and one mandibular premolar from each subject. The teeth from five patients, from two patients, and from three patients out of total 10 patients were used for cDNA microarray assay, for RT-PCR, and for real-time PCR, respectively. One premolar was used immediately after extraction (control), and another premolar was stored in liquid nitrogen at -196 degrees C for 1 week (cryopreserved) from each patient. PDL cells from these teeth were cultured separately through three passages. Total RNA was isolated and gene expression was compared between the cells from control and cryopreserved group out of each subject. The microarray data were validated using the reverse transcription-polymerase chain reaction (RT-PCR) and confirmed by quantitative real-time PCR. The cultured PDL cells from the control and cryopreserved teeth were of similar appearance under an optical microscope. In all subjects the fibroblast growth factor receptor 2 (FGFR2) gene was downregulated in the cells from the cryopreserved tooth. This study shows that cryopreservation of teeth affects the expression of the FGFR2 gene in cultured PDL cells, which is related to cell growth, cell development, and cell-cell signaling.


Assuntos
Criopreservação , Perfilação da Expressão Gênica , Ligamento Periodontal/citologia , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/biossíntese , Dente/fisiologia , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa
20.
Front Immunol ; 11: 553715, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33362756

RESUMO

CD137, a member of the TNFR family, is a costimulatory receptor, and CD137L, a member of the TNF family, is its ligand. Studies using CD137- and CD137L-deficient mice and antibodies against CD137 and CD137L have revealed the diverse and paradoxical effects of these two proteins in various cancers, autoimmunity, infections, and inflammation. Both their cellular diversity and their spatiotemporal expression patterns indicate that they mediate complex immune responses. This intricacy is further enhanced by the bidirectional signal transduction events that occur when these two proteins interact in various types of immune cells. Here, we review the biology of murine CD137/CD137L, particularly, the complexity of their proximal signaling pathways, and speculate on their roles in immune responses.


Assuntos
Ligante 4-1BB/imunologia , Transdução de Sinais/imunologia , Membro 9 da Superfamília de Receptores de Fatores de Necrose Tumoral/imunologia , Animais , Camundongos
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