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1.
BMC Oral Health ; 24(1): 390, 2024 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-38539170

RESUMO

BACKGROUND: Periodontal ligament stem cells (PDLSCs) have been proposed as therapeutic candidates in periodontal diseases and periodontium defects. Paracrine factors of PDLSCs, namely, secretome, can contribute to tissue regeneration comparable to direct stem cell application. This study explored restoration effects of PDLSC-derived secretome/conditioned medium (PDLSC-CM) on PDLSCs themselves in an inflammatory microenvironment and identified its action mechanisms using proteomics and transcriptomic profiling. METHODS: PDLSC-CM was prepared from cells under healthy culture conditions. Mass spectrometry and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were then performed to analyze the PDLSC-CM proteome. Osteogenic differentiation of PDLSCs under inflammatory conditions or in the presence of PDLSC-CM was then characterized in assays of alkaline phosphatase activity, intracellular calcium levels, protein expression of osteogenic markers, and matrix mineralization. Furthermore, the transcriptomic profile was assessed to identify significantly enriched signaling pathways and associated molecular networks by RNA sequencing. RESULTS: LC-MS/MS proteomics identified a total of 203 proteins and distinguished 187 significant protein changes in PDLSC-CM compared to control-CM. LPS-treated PDLSCs significantly attenuated osteogenic differentiation. When PDLSCs were treated with PDLSC-CM alone, their osteogenic activity was significantly upregulated compared to the control group. Moreover, the LPS-impaired osteogenesis of PDLSCs was reconstituted by PDLSC-CM treatment. RNA sequencing revealed 252, 1,326, and 776 differentially expressed genes in the control vs. LPS, control vs. PDLSC-CM, and LPS vs. LPS + PDLSC-CM groups, respectively. CONCLUSION: This study suggest that PDLSC-CM restores the osteogenic potential of PDLSCs in an inflammatory environment through secretory functions representing potential repair and regenerative mechanisms.


Assuntos
Ligamento Periodontal , Periodontite , Humanos , Osteogênese/genética , Meios de Cultivo Condicionados/farmacologia , Proteoma/farmacologia , Transcriptoma , Lipopolissacarídeos/farmacologia , Cromatografia Líquida , Secretoma , Espectrometria de Massas em Tandem , Células-Tronco , Diferenciação Celular , Células Cultivadas
2.
J Couns Psychol ; 70(5): 510-521, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37261794

RESUMO

We examined Asian American college students' adherence to traditional values that are salient in Asian cultures, the students' perceptions of their mother's and father's adherence to the same values, and the discrepancies between the students and their mothers and fathers on the levels of adherence to these values. Based on the data from 301 participants who self-identified as Asian Americans, paired-samples t tests revealed that the child-parent cultural value discrepancies were present across all generational statuses of the participants with the children adhering less strongly to most of the value dimensions than their parents. The results based on correlational analyses showed that many types of value discrepancies were positively associated with the likelihood and seriousness of conflict. Several types of value discrepancies also were inversely associated with the participants' life satisfaction and self-esteem. In addition, the results from the PROCESS Macro for mediation analysis revealed significant mediation role of family conflict on the relationships between various types of value discrepancies and life satisfaction. The significant mediators were the likelihood and seriousness of family conflict and the family conflict about education and career decisions, and the value discrepancies centered on the values of conformity to norms, family recognition through achievement, and humility. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Assuntos
Asiático , Cultura , Relações Familiares , Satisfação Pessoal , Autoimagem , Valores Sociais , Feminino , Humanos , Asiático/psicologia , Conflito Familiar/etnologia , Conflito Familiar/psicologia , Mães/psicologia , Pais/psicologia , Relações Pais-Filho/etnologia , Valores Sociais/etnologia , Estudantes/psicologia , Universidades , Pai/psicologia , Relações Familiares/etnologia , Relações Familiares/psicologia , Identificação Social
3.
J Couns Psychol ; 70(2): 203-211, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36521120

RESUMO

A body of research has been dedicated to demonstrating the relationship of perfectionism with a range of mental health indicators. Self-critical perfectionism, a component of perfectionism, has been framed primarily in a negative light within the mental health context. Given that research informs educational and clinical practices, it is important to explore the degree to which such findings generalize across cultures and subcultures. The current meta-analytic research systemically collated studies conducted with Asian college students with a particular attention to exploring whether orientation to Asian culture and cultural values correspondingly moderates the relationship between self-critical perfectionism and depressive symptoms. The degree of upholding Asian cultural values was represented by group identity (i.e., Asian American and Asian international). Eleven studies (N = 3,239) were identified through the literature search. Findings from the random-effects meta-analysis indicated a significant relationship between self-critical perfectionism and depressive symptoms in the overall sample. The group identity significantly moderated the relationship; among Asian international college students, self-critical perfectionism appeared to have a less harmful effect on mental distress compared to Asian American college students. Self-cultivation-one of the salient and virtuous Asian cultural values that aligns with self-critical perfectionism-may have motivated continuous striving for self-improvement to fulfill the honorable duty for their family for Asian international students. Additional findings and implications of the study are further discussed. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Assuntos
Asiático , Perfeccionismo , Humanos , Depressão/psicologia , Estudantes/psicologia , Escolaridade
4.
Int J Mol Sci ; 23(4)2022 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-35216377

RESUMO

We investigated the immune response mechanisms under systemic and local inflammation using mouse models whereby lipopolysaccharide (LPS) was administered intraperitoneally to induce systemic inflammation, and epicutaneous sensitization with ovalbumin was used to induce local inflammation. LPS increased the immune cell infiltration in the cardiac muscle near the aorta, alveoli, hepatic sinusoid, renal interstitium, and the submucosal layer of the duodenum. Similarly, ovalbumin increased the abundance of macrophages in the skin. Both LPS and ovalbumin induced NF-κB p65 and IκBα phosphorylation, as well as the expression of NF-κB target genes (TLR4, IL6, and TNFα). Additionally, both LPS and ovalbumin led to an increase in the absolute IL-1ß, IL-6, and TNFα serum levels and cytokine-related janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) phosphorylation. Moreover, the activated JAK2/STAT3 signaling increased the number of Ki67-positive cells (proliferating cells) and development pathway target gene expression (regeneration) in the inflammation models. In conclusion, LPS and ovalbumin increase immune cell infiltration in tissues, NF-κB activation, cytokine levels in serum, cytokine-stimulated JAK2/STAT3 signaling, and tissue regeneration.


Assuntos
Citocinas/metabolismo , Inflamação/metabolismo , Janus Quinase 2/metabolismo , Fator de Transcrição STAT3/metabolismo , Animais , Lipopolissacarídeos/metabolismo , Macrófagos/metabolismo , Camundongos , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais/fisiologia , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
5.
J Relig Health ; 59(2): 700-713, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29705827

RESUMO

This study presents the development and initial psychometric evaluation of the Religious Discrimination Scale (RDS). This 11-item instrument identified three dimensions based on perceived discrimination experiences of members of The Church of Jesus Christ of Latter-day Saints (LDS): Perceived Prejudice, Closet Symptoms, and Negative Labels. The psychometric evaluations of the RDS indicated a strong and clear factor structure as well as good internal consistency reliability. The test of measurement and structural invariance across gender also suggested that the RDS scale is equally appropriate to be used with both men and women. Implications for practice and research as well as future directions are discussed.


Assuntos
Discriminação Psicológica , Preconceito , Psicometria/estatística & dados numéricos , Inquéritos e Questionários/normas , Igreja de Jesus Cristo dos Santos dos Últimos Dias , Feminino , Humanos , Masculino , Reprodutibilidade dos Testes
6.
J Couns Psychol ; 66(2): 170-183, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30589282

RESUMO

The demand for high quality engineers is of particular importance as engineering jobs are projected to grow in the next 10 years (United States Bureau of Labor Statistics, 2018). More work is needed to understand factors related to academic engagement, satisfaction, and persistence intentions of Latino/as and women in engineering: 2 underrepresented groups in the engineering pipeline. We present findings that explored the role of social-cognitive, environmental, and personality variables in engineering persistence intentions, engagement and satisfaction of a diverse sample of 1,335 engineering students using an extension of the integrative social cognitive career theory model (SCCT; Lent et al., 2013). Results indicated that (a) the hypothesized model fit the data well for the full sample and across 8 subsamples based on gender-ethnicity (i.e., Latinas, Latinos, White women, and White men) and ethnicity-school type (i.e., Latina/os at Hispanic-serving institutions [HSIs], Latina/os at predominantly White institutions [PWIs], Whites at HSIs, and Whites at PWIs), (b) all but 5 model parameters were significant and positive for the full sample, (c) a subset of model parameters differed by the interactions of race/ethnicity-gender and race/ethnicity-school type groups, and (d) the relations within the model explained a significant amount of variance in engineering academic engagement, satisfaction, and persistence intentions for the full sample and 8 subsamples. Implications of the findings for educational and career interventions aimed at retaining Latina/os and women in engineering are discussed in relation to building on social cognitions in engineering academic engagement, satisfaction, and persistence intentions. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Assuntos
Sucesso Acadêmico , Cognição/fisiologia , Engenharia/educação , Intenção , Satisfação Pessoal , Estudantes/psicologia , Adulto , Feminino , Previsões , Humanos , Masculino , Autoeficácia , Comportamento Social , Estados Unidos/etnologia , Adulto Jovem
7.
J Relig Health ; 57(6): 2207-2223, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28470447

RESUMO

In this study, the Perceived Perfectionism from God Scale (PPGS) was developed with Latter-day Saints (Mormons) across two samples. Sample 1 (N = 421) was used for EFA to select items for the Perceived Standards from God (5 items) and the Perceived Discrepancy from God (5 items) subscales. Sample 2 (N = 420) was used for CFA and cross-validated the 2-factor oblique model as well as a bifactor model. Perceived Standards from God scores had Cronbach alphas ranging from .73 to .78, and Perceived Discrepancy from God scores had Cronbach alphas ranging from .82 to .84. Standards from God scores were positively correlated with positive affect, whereas Discrepancy from God scores was positively correlated with negative affect, shame and guilt. Moreover, these two PPGS subscale scores added significant incremental variances in predicting associated variables over and above corresponding personal perfectionism scores.


Assuntos
Perfeccionismo , Psicometria/instrumentação , Vergonha , Espiritualidade , Humanos , Religião
8.
J Cell Physiol ; 229(10): 1557-68, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24591095

RESUMO

The aim of this study is to determine whether GlcN could recover the endoplasmic reticulum (ER) stress-induced dysfunction of Na(+) /glucose cotransporter (SGLT) in renal proximal tubule cells (PTCs) under hypoxia. With the rabbit model, the renal ischemia induced tubulointerstitial abnormalities and decreased SGLTs expression in tubular brush-border, which were recovered by GlcN. Thus, the protective mechanism of GlcN against renal ischemia was being examined by using PTCs. Hypoxia decreased the level of protein O-GlcNAc and the expression of O-GlcNAc transferase (OGT) while increased O-GlcNAcase (OGA) and these were reversed by GlcN. Hypoxia also decreased the expression of SGLTs (SGLT1 and 2) and [(14) C]-α-methyl-D-glucopyranoside (α-MG) uptake which were recovered by GlcN and PUGNAc (OGA inhibitor). Hypoxia enhanced reactive oxygen species (ROS) and then ER stress proteins, glucose-regulated protein 78 (GRP78), and C/EBP-homologous protein (CHOP). However, the expression of GRP78 increased till 6 h and then decreased whereas CHOP increased gradually. Moreover, decreased GRP78 and increased CHOP were reversed by NAC (antioxidant) and GlcN. GlcN ameliorated hypoxia-induced decrease of O-GlcNAc modification of Sp1 but OGT or Sp1 siRNAs blocked the recovery effect of GlcN on SGLT expression and α-MG uptake. In addition, hypoxia-decreased GRP78 and HIF-1α expression was reversed by GlcN but OGT siRNA or Sp1 siRNA ameliorated the effect of GlcN. When PTCs were transfected with GRP78 siRNA or HIF-1α siRNA, SGLT expression and α-MG uptake was decreased. Taken together, these data suggest that GlcN-induced O-GlcNAc modified Sp1 with stimulating GRP78 and HIF-1α activity ameliorate hypoxia-induced SGLT dysfunction in renal PTCs. J. Cell. Physiol. 229: 1557-1568, 2014. © 2014 Wiley Periodicals, Inc.


Assuntos
Injúria Renal Aguda/tratamento farmacológico , Glucosamina/farmacologia , Isquemia/tratamento farmacológico , Túbulos Renais Proximais/efeitos dos fármacos , Transportador 1 de Glucose-Sódio/metabolismo , Transportador 2 de Glucose-Sódio/metabolismo , Fator de Transcrição Sp1/metabolismo , Injúria Renal Aguda/genética , Injúria Renal Aguda/metabolismo , Animais , Hipóxia Celular , Células Cultivadas , Modelos Animais de Doenças , Chaperona BiP do Retículo Endoplasmático , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Glicosilação , Proteínas de Choque Térmico/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Isquemia/genética , Isquemia/metabolismo , Túbulos Renais Proximais/metabolismo , Masculino , Metilglucosídeos/metabolismo , N-Acetilglucosaminiltransferases/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Cultura Primária de Células , Interferência de RNA , Coelhos , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição Sp1/genética , Fatores de Tempo , Fator de Transcrição CHOP/metabolismo , Transfecção , beta-N-Acetil-Hexosaminidases/metabolismo
9.
Cell Transplant ; 33: 9636897231217382, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38229498

RESUMO

Because there is a shortage of donor kidneys, researchers are exploring the possibility of using genetically modified pig kidneys for transplantation. Approaches involving knockout of carbohydrate genes or knockin of protective proteins have been attempted to determine the best gene modifications. In this study, we utilized GalT-/-;hCD39;hCD55 and GalT-/-;hCD39;hCD46;hCD55;thrombomodulin (TBM) pigs for transplantation in nonhuman primates (NHPs). The NHPs survived for 4 weeks after kidney transplantation (4 WAT) from the GalT-/-;hCD39;hCD55 pig and for 6 WAT from the GalT-/-;hCD39;hCD46;hCD55;TBM pig. However, messenger RNA (mRNA) sequencing and immunohistochemistry analysis revealed that the 6 WAT kidney exhibited more severe apoptosis, inflammation, loss of renal function, and renal fibrosis than the 4 WAT kidney. These results indicate that additional knockin of complement regulator (hCD46) and coagulation regulator (TBM) is not enough to prevent renal damage, suggesting that improved immune suppression is needed for more prolonged survival.


Assuntos
Transplantes , Animais , Suínos , Animais Geneticamente Modificados , Transplante Heterólogo/métodos , Primatas , Rim , Rejeição de Enxerto
10.
ACS Appl Mater Interfaces ; 16(24): 30611-30621, 2024 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-38857116

RESUMO

Polypyrrole (Ppy) is a biologically compatible polymer that is used as a matrix, in which drugs and enzymes can be incorporated by doping. Here, we suggest an inventive application of Ppy as a biorecognition film encapsulated with an antibody (Ab) as an alternative strategy for the on-site multistep functionalization of thiol-based self-assembled monolayers. The fabrication steps of the recognition films were followed by dropping pyrrole and Ab mixed solutions onto the electrode and obtaining a thin film by direct current electropolymerization. The efficiency of Ab immobilization was studied by using fluorescence microscopy and electrochemical (EC) methods. Finally, the Ab density was increased and immobilized in 1 min, and the sensing performance as an EC immunosensor was demonstrated using α-fetoprotein with a limit of detection of 3.13 pg/mL and sensing range from 1 pg/mL to 100 ng/mL. This study demonstrates the potential for electrochemical functionalization of biomolecules with high affinity and rapidity.


Assuntos
Anticorpos Imobilizados , Técnicas Eletroquímicas , Polímeros , Pirróis , Pirróis/química , Imunoensaio/métodos , Polímeros/química , Técnicas Eletroquímicas/métodos , Anticorpos Imobilizados/imunologia , Anticorpos Imobilizados/química , Técnicas Biossensoriais/métodos , Polimerização , alfa-Fetoproteínas/análise , alfa-Fetoproteínas/imunologia , Eletrodos , Limite de Detecção , Humanos
11.
Behav Sci (Basel) ; 13(7)2023 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-37504023

RESUMO

The purpose of this study was to test if perceived social support and self-compassion will interact to reduce the magnitude of the bivariate relationship (buffering effect) between cumulative trauma and trauma symptoms after controlling for gender and age among college students. As part of a broader research project conducted between 2018 and 2019, we collected data via online surveys from a sample of 551 undergraduate students at a public university in the southern region of the US. After data cleaning, the study included 538 participants (representing 97.6% of the original dataset), ensuring a diverse representation across various ethnicities and genders. The three-way interaction model accounted for 38.61% of the variance in PTSD symptoms. In detail, with high levels of perceived social support, there was a significant difference in the buffering effects of perceived social support on the trauma-PTSD association between high and low self-compassion. Conversely, at high levels of self-compassion, perceived social support did not significantly influence the buffering effect of self-compassion. This study underscores the critical role of self-compassion in enhancing the protective effect of high-level perceived social support against PTSD symptoms following cumulative trauma.

12.
ACS Omega ; 8(1): 1389-1400, 2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36643538

RESUMO

Gold is of considerable interest for electrochemical active surfaces because thiol-modified chemicals and biomolecules can be easily immobilized with a simple procedure. However, most gold surfaces are damaged with repetitive measurements, so they are difficult to reuse. Here we demonstrate a novel electrochemical cleaning method of gold surfaces to reuse electrodes with a simple protocol that is easy and nontoxic. This electrochemical cleaning consists of two steps by using different solutions. The 1st step is a cyclic voltammetry sweep using a very low concentration of sulfuric acid, and the 2nd step is a cyclic voltammetry sweep using potassium ferricyanide. Different cleaning methods were also considered for comparison. Consequently, after assembling and desorption of the cell and antigen, the changes in gold electrode performance, as immunosensor and cytosensor, were investigated by electrochemical impedance and cyclic voltammetry. It was found that repetitive measurement is possible until five times while maintaining the reproducibility. It is believed that this method is capable of enabling reuse of gold electrodes and can be used for long-term and accurate monitoring of biological effects, especially at a low cost.

13.
Toxicol Res ; 39(4): 601-609, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37779585

RESUMO

We investigated the cytotoxic effect of Pelargonium sidoides extract on Madin-Darby canine kidney (MDCK) cells. P. sidoides extract decreased the cell viability in a dose dependent manner (> 0.2%). The extract of P. sidoides decreased the mitochondrial action potential, increased the number of reactive oxygen species (ROS) inside the cell, and caused nicotinamide adenine dinucleotide hydride (NADH) to be released, all of which are signs of mitochondrial dysfunction. The results of unbiased mRNA sequencing showed that 0.3% P. sidoides extract upregulates the apoptosis-related gene (BBC3). This finding was supported by immunoblot analysis of apoptosis signal pathways, which included Bcl-2, Bax, cytochrome C (CytC), cleaved caspase 3 (CC3), cleaved caspase 7 (CC7), cleaved caspase 9 (CC9) and cleaved PARP (CP). It is interesting to note that the elevated levels of Bax, CytC, CC3, CC7, and CC9, as well as CP, were suppressed by N-acetyl-L-cysteine (NAC) pretreatment, which points to ROS-mediated apoptosis. The small GTPases, RhoA, and Rac1/cdc42-GTP-bound active form were all lowered when P. sidoides extract was used. Also, RhoA-related cytoskeleton signals (ROCK, p-LIMK1/2, p-cofilin) and Rac1/cdc42-related signals (N-WASP, WAVE-2) were inhibited by P. sidoides extract. NAC or RhoA/Rac1/cdc42 activator pretreatment reduced P. sidoides extract-induced actin destabilization. In this work, P. sidoides extract promotes apoptosis by causing mitochondrial dysfunction and cytoskeleton disassembly.

14.
J Cell Physiol ; 227(4): 1731-9, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21688265

RESUMO

Stem cell functions are dramatically altered by oxygen in tissue culture, which means the antioxidant/oxidant balance is critical for protection as well as toxicity. This study examined the effect of the heparin-binding growth factor midkine (MK) on hypoxia-induced apoptosis and related signal pathways in mouse embryonic stem cells (mESCs). Hypoxia (60 h) increased lactate dehydrogenase release and apoptosis, and reduced cell viability and proliferation. These effects were reversed by MK (100 ng/ml). MK also reversed hypoxia-induced increases of intracellular reactive oxygen species, c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK) phosphorylation. Blockage of JNK and p38 MAPK using small interference (si)RNAs produced a decrease in apoptosis. A loss of mitochondrial membrane potential, increases of cytochrome c release from mitochondria to cytosol, and cleaved caspase-3 expression, as well as decreases in cIAP-2 and Bcl-2 were also reversed by MK. Hypoxia alone and hypoxia with MK increased low-density lipoprotein receptor-related protein-1 (LRP-1) mRNA and protein expression. Hypoxia with MK rapidly increased serine/threonine protein kinase (Akt) phosphorylation which reversed by LRP-1 Ab (0.1 µg/ml) and prolonged heme oxygenase-1 (HO-1) expression. In addition, hypoxia with MK increased the expression of hypoxia-inducible factor-1α (HIF-1α). Moreover, inhibition of Akt, HO-1, and HIF-1α signaling pathways abolished the MK-induced blockage of apoptosis. In conclusion, MK partially prevented hypoxic injury of mESCs through activation of Akt, HO-1, and HIF-1α via LRP-1.


Assuntos
Hipóxia Celular/efeitos dos fármacos , Citocinas/farmacologia , Células-Tronco Embrionárias/efeitos dos fármacos , Células-Tronco Embrionárias/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores de LDL/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Animais , Apoptose/efeitos dos fármacos , Células Cultivadas , Citoproteção/efeitos dos fármacos , Células-Tronco Embrionárias/citologia , Heme Oxigenase-1/metabolismo , Proteína-1 Relacionada a Receptor de Lipoproteína de Baixa Densidade , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Camundongos , Midkina , Modelos Biológicos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos
15.
Vet Med Sci ; 8(6): 2422-2433, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36040758

RESUMO

BACKGROUND: Bentonite, a montmorillonite clay, has been used as a classical remedy strategy for a long time. Recently, bentonite has been used as a raw material in cosmetic products because of its antibacterial and antioxidant properties. However, the therapeutic effect of bentonite on burn injuries has not yet been identified. OBJECTIVES: The aim of this study was to explore the therapeutic effect of a novel bentonite complex, which was developed for medical use, on burn wounds and the anti-inflammatory function of bentonite clay in the complex in vitro. METHODS: A novel bentonite complex and bentonite clay were prepared for each in vivo and in vitro assay (C&L Biotech, Seoul, Korea). Burn wounds were induced on the dorsal skin of two Yucatan minipigs, and effects of tissue regeneration and anti-inflammation were assessed by histological and gene expression analysis. RESULTS: The bentonite complex improved skin regeneration in burn wounds by inducing collagen synthesis, cell proliferation and angiogenesis in vivo. Furthermore, expression of inflammatory cytokines was significantly inhibited by treatment of the bentonite complex. In vitro study for identification of anti-inflammatory effect showed that bentonite clay significantly regulated COX-2 signalling in both HacaT keratinocytes and 3D4/2 macrophage cell lines. CONCLUSIONS: This study identified the therapeutic effect of a novel bentonite complex in burn wounds by inducing skin regeneration and bentonite-mediated anti-inflammatory responses.


Assuntos
Antibacterianos , Bentonita , Animais , Suínos , Bentonita/farmacologia , Bentonita/uso terapêutico , Argila , Porco Miniatura , Antioxidantes
16.
Biosensors (Basel) ; 12(2)2022 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-35200367

RESUMO

Nicotinamide adenine dinucleotide (NADH) is an important cofactor involved in metabolic redox reactions in living cells. The detection of NADH in living animal cells is a challenge. We developed a one-step monitoring method for NADH via an electrocatalytic reaction that uses a surface-modified, screen-printed electrode (SPE) having a redox active monolayer 4'-mercapto-N-phenlyquinone diamine (NPQD) formed by a self-assembled monolayer (SAM) of an aromatic thiol, 4-aminothiophenol (4-ATP). This electrode has a limit of detection (LOD) of 0.49 µM and a sensitivity of 0.0076 ± 0.0006 µM/µA in cell culture media, which indicates that it retains its selectivity. The applicability of this NADH sensor was demonstrated for the first time by cell viability monitoring via NADH-sensing in cell culture supernatants.


Assuntos
Técnicas Biossensoriais , NAD , Animais , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas , Eletrodos , Camundongos , Oxirredução
17.
Sci Rep ; 12(1): 16716, 2022 10 06.
Artigo em Inglês | MEDLINE | ID: mdl-36202932

RESUMO

Monitoring nicotinamide adenine dinucleotide (NADH) is important because NADH is involved in cellular redox reactions and cellular energy production. Currently, few biosensors quantify NADH in whole blood. However, they still have limitations due to several defects, including poor repeatability, long analysis time, and their requirement of extra sample pretreatment. In this study, we developed electrocatalytic sensors using screen-printed electrodes with a redox-active monolayer 4'-mercapto-N-phenylquinone diamine formed by a self-assembled monolayer of a 4-aminothiophenol (4-ATP). We exhibited their behavior as electrocatalysts toward the oxidation of NADH in whole blood. Finally, the electrocatalytic sensors maintained stability and exhibited 3.5 µM limit of detection, with 0.0076 ± 0.0006 µM/µA sensitivity in a mouse's whole blood. As proof of concept, a polyhexamethylene guanidine phosphate-treated mouse model was used to induce inflammatory and fibrotic responses, and NADH level was measured for 45 days. This work demonstrates the potential of electrocatalytic sensors to analyze NADH in whole blood and to be developed for extensive applications.


Assuntos
Técnicas Biossensoriais , NAD , Trifosfato de Adenosina , Animais , Diaminas , Eletroquímica , Eletrodos , Camundongos , NAD/análise , Oxirredução
18.
Lab Anim Res ; 38(1): 32, 2022 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-36266672

RESUMO

BACKGROUND: Given its similar structure and immune response to the human skin, porcine is a good model for dermal studies. Here, we sensitized ovalbumin (Ova) on minipig back skin for 2-4 weeks to induce chronic atopic dermatitis (AD). RESULTS: Gross observation, serum cytokine level, epidermal thickness, and epidermal integrity did not change after 4 weeks of Ova induction compared with the control, indicating AD modeling failure. Only the neutrophils in the blood and macrophages in bronchoalveolar lavage fluid changed slightly until 3 or 2 weeks after Ova sensitization, respectively. The successful and failed Ova-induced AD minipig models only differ in age and body weight of the minipigs. The minipigs, 12 months old with a 30-kg median weight, had a two-fold thicker dermis than minipigs 8-10 months old, with an 18.97-kg median weight, resulting in impaired Ova permeability and immune response. CONCLUSION: Age and body weight are key factors that should be considered when developing an Ova-induced AD minipig model.

19.
J Cell Physiol ; 226(12): 3422-32, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21344393

RESUMO

Adhesion of cells to extracellular matrix (ECM) influences vital aspects of anchorage-dependent cell behavior including survival, proliferation, and differentiation. However, the role of collagen I in mouse embryonic stem cells (mESCs) is not well-known. Therefore, in the present study we examined the effect of collagen I on mESC self-renewal and related signal pathways. Collagen I (10 µg/ml) maintained mESCs in an undifferentiated state (Nanog, OCT4, and SSEA-1) and did not affect differentiation (GATA4, Tbx5, Fgf5, and Cdx2) in the presence of leukemia inhibitory factor (LIF). Treatment with collagen I bound both α2ß1 integrin and discoidin domain receptor 1 (DDR1), and stimulated intracellular signaling pathways. Collagen I-bound α2ß1 integrin increased integrin-linked kinase (ILK) phosphorylation, cleaved Notch protein expression in the nuclear fraction, and Gli-1 mRNA expression. In addition, collagen I-bound DDR1 increased GTP-bound Ras, phosphoinositide 3-kinase (PI3K) p85α catalytic subunit protein expression, and Akt and ERK phosphorylation. Importantly, collagen I increased Bmi-1 protein expression in the nucleus which was blocked by small interfering RNA (siRNA) specific for Gli-1 and ERK, showing that parallel pathways of integrins and DDR1 merge at Bmi-1. Furthermore, collagen I-induced p16 decrease and p-Rb increase were reversed by Bmi-1-specific siRNA. Moreover, Bmi-1 silencing abolished the collagen I-induced increase of proliferation indices and undifferentiation markers. These results indicate that collagen I stimulates the self-renewal of mESCs mediated by Bmi-1 through α2ß1 integrin-dependent ILK, Notch, Gli-1, and DDR1-dependent Ras, PI3K/Akt, and ERK.


Assuntos
Proliferação de Células , Colágeno Tipo I/metabolismo , Células-Tronco Embrionárias/enzimologia , Integrina alfa2beta1/metabolismo , Proteínas Nucleares/metabolismo , Células-Tronco Pluripotentes/enzimologia , Proteínas Proto-Oncogênicas/metabolismo , Receptores Proteína Tirosina Quinases/metabolismo , Proteínas Repressoras/metabolismo , Transdução de Sinais , Animais , Adesão Celular , Proteínas de Ciclo Celular/metabolismo , Linhagem Celular , Receptor com Domínio Discoidina 1 , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Fatores de Transcrição Kruppel-Like/metabolismo , Camundongos , Proteínas Nucleares/genética , Fosfatidilinositol 3-Quinase/metabolismo , Fosforilação , Complexo Repressor Polycomb 1 , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Interferência de RNA , RNA Mensageiro/metabolismo , Receptores Notch/metabolismo , Proteínas Repressoras/genética , Transdução de Sinais/genética , Fatores de Tempo , Proteína GLI1 em Dedos de Zinco , Proteínas ras/metabolismo
20.
PLoS One ; 16(6): e0252947, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34086835

RESUMO

Minipigs have remarkably similar physiology to humans, therefore, they it can be a good animal model for inflammation study. Thus, the conventional (serum chemistry, histopathology) and novel analytic tools [immune cell identification in tissue, cytokine level in peripheral blood mononuclear cells (PBMC) and serum, NF-kB target gene analysis in tissue] were applied to determine inflammation in Chicago Miniature Swine (CMS) minipig. Lipopolysaccharide (LPS)-induced acute systemic inflammation caused liver and kidney damage in serum chemistry and histopathology. Immunohistochemistry (IHC) also showed an increase of immune cell distribution in spleen and lung during inflammation. Moreover, NF-kB-target gene expression was upregulated in lung and kidney in acute inflammation and in heart, liver, and intestine in chronic inflammation. Cytokine mRNA was elevated in PBMC under acute inflammation along with elevated absolute cytokine levels in serum. Overall, LPS-mediated systemic inflammation affects the various organs, and can be detected by IHC of immune cells, gene analysis in PBMC, and measuring the absolute cytokine in serum along with conventional inflammation analytic tools.


Assuntos
Citocinas/sangue , Regulação da Expressão Gênica , Inflamação/imunologia , Leucócitos Mononucleares/imunologia , NF-kappa B/metabolismo , Animais , Modelos Animais de Doenças , Perfilação da Expressão Gênica , Inflamação/sangue , Inflamação/induzido quimicamente , Inflamação/genética , Leucócitos Mononucleares/metabolismo , Lipopolissacarídeos/toxicidade , NF-kappa B/genética , Suínos , Porco Miniatura
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