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1.
J Zoo Wildl Med ; 50(4): 927-936, 2020 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-31926525

RESUMO

Seabirds have been widely used for monitoring the health of the oceans in diverse marine regions. Among low-cost survey strategies, systematic surveys of seabirds beached on coasts have been developed since the 20th century. However, these studies do not always address blood aspects. The assessment of the health status of birds based on the analysis of hematological and plasma chemistry is crucial to evaluate the overall health status profile of live organisms. Here, the authors study the variability of blood parameters by sex, age class, and year of beached Magellanic Penguin during the nonreproductive period in northern Argentina. Of 44 penguins, 77% were categorized as younger juveniles and the rest as older juveniles, and were captured and studied in coastal areas of Buenos Aires Province during the summers of 2017 and 2018. The mean body weight of beached penguins was affected by the age class of the individuals; most of the younger juveniles showed poor condition in terms of body mass (1,761 ± 235 g). No significant differences were observed in body weight between years and sex. Still, there were significant differences between years for alkaline phosphatase (ALP) and creatine phosphokinase (CPK) values. Twelve of the 20 blood parameters analyzed differ significantly with the age class of the beached penguins; younger juveniles were in a state of inanition. Our results may serve as a necessary first step in improving the conservation status of the Magellanic Penguin in nonbreeding grounds of Argentina, and call for a better knowledge of the health status of the species along its annual cycle.


Assuntos
Contagem de Eritrócitos/veterinária , Hematócrito , Contagem de Leucócitos/veterinária , Estações do Ano , Spheniscidae/sangue , Envelhecimento , Fosfatase Alcalina/sangue , Animais , Animais Selvagens , Anticorpos Heterófilos , Argentina , Glicemia , Proteínas Sanguíneas , Colesterol/sangue , Creatina Quinase/sangue , Feminino , Hemoglobinas , Contagem de Linfócitos/veterinária , Masculino , Fósforo/sangue , Transaminases/sangue , Ureia/sangue , Ácido Úrico/sangue
2.
J Zoo Wildl Med ; 50(4): 1021-1025, 2020 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-31926541

RESUMO

Hematology and serum biochemistry profiles are used to evaluate the health status of animals ongoing rehabilitation. The aim of this project was to develop blood and biochemistry ranges for harbor seal pups (Phoca vitulina) after rehabilitation; thus, 22 different blood parameters in 60 animals were tested before release. The second goal was to test for differences due to sex, stranding location, body condition at admission, and presence or absence of umbilical cord. The alanine aminotransferase, ALT (or glutamate pyruvate transaminase, GPT), (ALT-GPT) differed significantly (P bq = 0.00851) between sexes. Lower leukocyte counts and higher liver enzyme values were the most remarkable findings when comparing the results of this study to other published data. This is the first study to report blood reference ranges for harbor seal pups in the Dutch Wadden Sea after rehabilitation.


Assuntos
Contagem de Eritrócitos/veterinária , Hematócrito/veterinária , Contagem de Leucócitos/veterinária , Phoca/sangue , Contagem de Plaquetas/veterinária , Alanina Transaminase/sangue , Fosfatase Alcalina/sangue , Animais , Glicemia , Nitrogênio da Ureia Sanguínea , Creatinina , Feminino , Hemoglobinas , Masculino
3.
Medicine (Baltimore) ; 99(4): e18788, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31977869

RESUMO

BACKGROUND: This systematic review and meta-analysis were performed to evaluate kidney function in patients with sepsis-associated acute kidney injury (SA-AKI) on alkaline phosphatase (AP) therapy. METHODS: PubMed, Embase, and the Cochrane Central Register of Controlled Trials were searched electronically from inception until May 4, 2019 and randomized controlled studies assessing AP treatment in patients with SA-AKI were included. Pool analyses with fixed effects or random effects models calculated pooled mean, standard deviation, and odds ratio (OR) with 95% confidence interval (CI). RESULTS: Four randomized controlled trials involving AP therapy for 392 patients with SA-AKI were included. AP had a positive effect on endogenous creatinine clearance (ECC) in patients with SA-AKI at day 14 (random effects: mean difference = 10.56, 95% CI = 2.27-18.84, P = .01) and day 28 (random effects: mean difference = 14.30, 95% CI = 6.27-22.33, P = .0005). All-cause mortality at day 28 (fixed effects: OR = 0.62, 95% CI = 0.40-0.97, P = .04) and day 90 (fixed effects: OR = 0.61, 95% CI = 0.39-0.96, P = .03) improved. Plasma creatinine level (fixed effects: mean difference = -76.83, 95% CI = -146.92 to -6.74, P = .03) and biomarkers level (random effects: mean difference = -6.57, 95% CI = -10.74 to -2.40, P < .00001) also improved in the therapy group compared with placebo. CONCLUSION: In patients with SA-AKI, AP showed a relatively late protective effect by improving ECC at days 7, 14, and 28. ECC level improved when patients received AP dose of 0.212 mg/kg. Mortality improved at days 28 and 90, respectively, when patients received AP dose of 1.6 mg/kg. Levels of overall AKI biomarkers were improved in short term.


Assuntos
Lesão Renal Aguda/tratamento farmacológico , Fosfatase Alcalina/administração & dosagem , Lesão Renal Aguda/etiologia , Lesão Renal Aguda/metabolismo , Fosfatase Alcalina/farmacologia , Creatinina/sangue , Relação Dose-Resposta a Droga , Humanos , Ensaios Clínicos Controlados Aleatórios como Assunto , Sepse/complicações
4.
Medicine (Baltimore) ; 99(2): e18545, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31914029

RESUMO

This study aimed to determine the impact of the neutrophil-to-lymphocyte ratio (NLR) and the platelet-to-lymphocyte ratio (PLR) on the prognosis of nasopharyngeal carcinoma (NPC) before and after intensity modulated radiotherapy (IMRT).Pre/post-treatment and changes in inflammatory biomarker levels of 207 patients who were diagnosed with NPC and received IMRT between January 2012 and December 2014 were analyzed, and the cellular biomarker analyses were from patient blood. ROC (receiver operating characteristic) analysis was used to decide the optimal cutoff values of NLR and changes in NLR (ΔNLR) and PLR (ΔPLR). The Kaplan-Meier and logarithmic rank methods were used to compare overall survival times between groups. Univariate analysis was used to investigate the effects of age, gender, histology, Karnofsky performance score (KPS), TNM stage, clinical stage, course of disease and lymphocyte, neutrophil and platelet counts as well as alkaline phosphatase (ALP) levels on the prognosis of NPC. The independent predictors of OS were determined by Cox multivariate regression analysis.The optimal cut-off values of NLR, PLR, ΔNLR and ΔPLR were 2.49, 155.82, 1.80, and 100.00, respectively. These were used to classify patients into high (NLR > 2.49) and low NLR groups (NLR < 2.49); high (PLR>155.82) and low (PLR < 155.82) PLR groups; high (ΔNLR>1.80) and low ΔNLR groups (ΔNLR < 1.80); high (ΔPLR > 100.00) and low ΔPLR groups (ΔPLR < 100.00). TNM stage, clinical stage and ALP levels were highly correlated with high NLR and PLR. Cox multivariate regression analysis suggested that the ΔNLR (HR = 2.89, 95% CI: 1.33∼2.78) was independent of the characteristics for NPC.As a novel inflammatory index, ΔNLR appears to have some predictive power for the prognosis of patients with NPC.


Assuntos
Linfócitos/efeitos da radiação , Carcinoma Nasofaríngeo/radioterapia , Neoplasias Nasofaríngeas/radioterapia , Neutrófilos/efeitos da radiação , Radioterapia de Intensidade Modulada/efeitos adversos , Adulto , Idoso , Idoso de 80 Anos ou mais , Fosfatase Alcalina/sangue , Contagem de Células Sanguíneas , Plaquetas/patologia , Plaquetas/efeitos da radiação , Feminino , Humanos , Linfócitos/patologia , Masculino , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo/mortalidade , Carcinoma Nasofaríngeo/patologia , Neoplasias Nasofaríngeas/mortalidade , Neoplasias Nasofaríngeas/patologia , Estadiamento de Neoplasias/métodos , Neutrófilos/patologia , Valor Preditivo dos Testes , Prognóstico , Radioterapia de Intensidade Modulada/métodos , Estudos Retrospectivos
5.
J Appl Oral Sci ; 28: e20190105, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31800873

RESUMO

Calcium aluminate cement (CAC) has been highlighted as a promising alternative for endodontic use aiming at periapical tissue repair. However, its effects on dental pulp cells have been poorly explored. OBJECTIVE: This study assessed the impact of calcium chloride (CaCl2) and bismuth oxide (Bi2O3) or zinc oxide (ZnO) additives on odontoblast cell response to CAC. METHODOLOGY: MDPC-23 cells were exposed for up to 14 d: 1) CAC with 2.8% CaCl2 and 25% ZnO (CACz); 2) CAC with 2.8% CaCl2 and 25% Bi2O3 (CACb); 3) CAC with 10% CaCl2 and 25% Bi2O3 (CACb+); or 4) mineral trioxide aggregate (MTA), placed on inserts. Non-exposed cultures served as control. Cell morphology, cell viability, gene expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), and dentin matrix protein 1 (DMP-1), ALP activity, and extracellular matrix mineralization were evaluated. Data were compared using ANOVA (α=5%). RESULTS: Lower cell density was detected only for MTA and CACb+ compared with Control, with areas showing reduced cell spreading. Cell viability was similar among groups at days one and three (p>0.05). CACb+ and MTA showed the lowest cell viability values at day seven (p>0.05). CACb and CACb+ promoted higher ALP and BSP expression compared with CACz (p<0.05); despite that, all cements supported ALP activity. Matrix mineralization were enhanced in CACb+ and MTA. CONCLUSION: In conclusion, CAC with Bi2O3, but not with ZnO, supported the expression of odontoblastic phenotype, but only the composition with 10% CaCl2 promoted mineralized matrix formation, rendering it suitable for dentin-pulp complex repair.


Assuntos
Compostos de Alumínio/química , Compostos de Alumínio/farmacologia , Compostos de Cálcio/química , Compostos de Cálcio/farmacologia , Cimentos Dentários/química , Cimentos Dentários/farmacologia , Polpa Dentária/citologia , Polpa Dentária/efeitos dos fármacos , Fosfatase Alcalina/análise , Fosfatase Alcalina/efeitos dos fármacos , Animais , Bismuto/química , Bismuto/farmacologia , Cloreto de Cálcio/química , Cloreto de Cálcio/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Combinação de Medicamentos , Expressão Gênica/efeitos dos fármacos , Teste de Materiais , Camundongos , Odontoblastos/efeitos dos fármacos , Óxidos/química , Óxidos/farmacologia , Reprodutibilidade dos Testes , Silicatos/química , Silicatos/farmacologia , Fatores de Tempo , Óxido de Zinco/química , Óxido de Zinco/farmacologia
6.
Toxicol Lett ; 319: 242-249, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31733320

RESUMO

Humans are typically exposed to mixtures of substances, whereby their bioactivity can be significantly altered by co-occurring compounds. Thus, over the last years, research on combinatory effects has gained increasing attention. In particular, several xenoestrogens have been recently reported to interact synergistically, among them alternariol (AOH) and zearalenone (ZEN), two toxins produced by molds which contaminate crops or food commodities. Bisphenol A (BPA) is a potential food contaminant arising from its use in plastics and represents a well-known xenoestrogen, acting as an endocrine disruptor. However, little research was yet conducted on its impact on the bioactivity of other xenoestrogens, and vice versa. Thus, in this study, we focused on combinatory estrogenic effects of BPA with AOH and ZEN in Ishikawa cells, which represent a well-established, estrogen-sensitive human cell model. Estrogenic stimuli of the single compounds and binary combinations in constant concentration ratios were measured by assessing the activity of alkaline phosphatase, a natural reporter gene for estrogen receptor activation. In parallel, cytotoxicity was monitored by neutral red assay. For statistical analysis of combinatory effects the "combination index" model was applied. In combination with ZEN, BPA was found to cause additive estrogenic effects. Mixtures of BPA with AOH expressed moderately antagonistic to nearly additive combinatory effects, depending on the concentration ratio. Although no synergistic effects were measured in the applied chemical mixtures, additive estrogenic stimuli were observed, underlining the importance to consider the cumulative impact of endocrine active factors out of different sources and structural classes.


Assuntos
Compostos Benzidrílicos/toxicidade , Endométrio/efeitos dos fármacos , Estrogênios/toxicidade , Lactonas/toxicidade , Micotoxinas/toxicidade , Fenóis/toxicidade , Zearalenona/toxicidade , Fosfatase Alcalina/análise , Fosfatase Alcalina/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Interações de Medicamentos , Disruptores Endócrinos/toxicidade , Endométrio/citologia , Feminino , Humanos
7.
Oral Dis ; 26(1): 131-144, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31541596

RESUMO

OBJECTIVE: Secondary alveolar bone grafting is an essential part in the treatment of alveolar cleft deformity. Autologous iliac bone is the most favorable grafting source. However, the factors regulating postoperative bone formation are unclear. Investigations are needed to found whether the alveolar bone niche and bone marrow mesenchymal stem cells (BMSCs) derived from the jaw bone (BMSCs-J) affected the osteogenesis of BMSCs from the ilium (BMSCs-I). MATERIALS AND METHODS: The effect of BMSCs-J on BMSCs-I was investigated using a co-culture model. The exosomes were purified by sequential centrifugation. The osteoblastic differentiation of BMSCs was analyzed in vitro and in vivo. RESULTS: Co-culture with BMSCs-J increased the alkaline phosphatase (ALP) activity, Alizarin Red S (ARS) staining, and osteogenic gene expression in BMSCs-I. Transmission electron microscopy and nanoparticle tracking analysis verified the presence of exosomes in the culture supernatants of BMSCs. Exosomes secreted by BMSCs-J enhanced the ALP activity, ARS staining, osteogenic gene expression of BMSCs-I in vitro, and new bone formation in vivo. Blocking the secretion of exosomes using siRNA for Rab27a inhibited the effect of BMSCs-J. CONCLUSION: Exosomes played a role in the interaction between BMSCs-J and BMSCs-I, thereby leading to the enhanced osteogenic capacity of BMSCs-I and bone formation.


Assuntos
Células da Medula Óssea/citologia , Exossomos/fisiologia , Ílio/citologia , Células-Tronco Mesenquimais/citologia , Osteogênese , Fosfatase Alcalina/metabolismo , Diferenciação Celular , Células Cultivadas , Técnicas de Cocultura , Humanos , Arcada Osseodentária/citologia
8.
Bioresour Technol ; 297: 122406, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31787513

RESUMO

The objective of this study was to evaluate changes in phosphorus fractions during vegetable waste and chicken manure composting. High throughput sequencing, quantitative PCR, and multiple analysis methods were applied to investigate interconnections among phosphorus fractions, enzyme activity, and phoD-harboring bacterial community composition. We found the highest composting temperature reached 61 °C and phosphorus fractions presented significant differences during a 60-day composting. The content of plant-absorbable phosphorus, including water soluble phosphorus, available phosphorus, and citric acid phosphorus increased by 121%, 87%, and 63%, respectively. Additionally, phoD gene abundance significantly correlated with the activities of nine enzymes. Our findings emphasize that microbial activity plays an important role in phosphorus transformation during composting, and the final composting product could be good biological phosphorus fertilizer. To our knowledge, this is the first report indicating that enzyme activity, community composition and abundance of phoD-harboring bacteria have direct and indirect effects on phosphorus transformation during composting.


Assuntos
Compostagem , Fosfatase Alcalina , Animais , Bactérias , Galinhas , Esterco , Fósforo , Solo , Verduras
9.
Gene ; 726: 144193, 2020 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-31669647

RESUMO

OBJECTIVE: miR-199a-5p was increased during osteoblast differentiation, which may target and regulate TET2, a gene attracted a lot of attention in the osteoblast differentiation in the past few years. However, the role of miR-199a-5p in osteoblast differentiation by targeting TET2 is not established. METHODS: The correlation between miR-199a-5p and TET2 was verified through dual luciferase reporter assay, and their expressions in human bone marrow stromal cells (hBMSCs) during the osteoblast differentiation were detected. hBMSCs were transfected with TET2 siRNA, miR-199a-5p mimic or/and TET2 CRISPR activation plasmid., and then prepared for the induction of osteoblast differentiation, followed by alkaline phosphatase (ALP) and alizarin red staining, qRT-PCR and Western blotting. In vivo, ovariectomized (OVX) mice were injected with agomir-miR-199a-5p, antagomiR-199a-5p or/and TET2 siRNA to calculate the BMD and BV/TV ratio of mice, as well as to measure the expressions of osteogenesis-related genes in bone tissues. RESULTS: A gradual increase of miR-199a-5p was observed in hBMSCs during the induction of osteoblast differentiation, while TET2 expression was decreased. Besides, miR-199a-5p was reduced in the bone tissue of OVX mice, while TET2 was up-regulated. In addition, overexpression of miR-199a-5p and inhibition of TET2 augmented ALP activity in hBMSCs, with the enhanced calcification and the up-regulated expressions of Runx2, OSX and OCN, which also increased the quality of bone in OVX mice accompanying the enhancement BV/TV ratio, BMD and osteogenesis-related genes. CONCLUSION: MiR-199a-5p may promote the osteoblast differentiation and prevent OVX-induced osteoporosis by targeting TET2.


Assuntos
Diferenciação Celular/genética , Proteínas de Ligação a DNA/genética , MicroRNAs/genética , Osteoblastos/metabolismo , Proteínas Proto-Oncogênicas/genética , Fosfatase Alcalina/genética , Animais , Células da Medula Óssea/metabolismo , Osso e Ossos/metabolismo , Calcificação Fisiológica/genética , Células Cultivadas , Feminino , Humanos , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Osteogênese/genética , Regulação para Cima/genética
10.
Arch Oral Biol ; 109: 104582, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31605918

RESUMO

OBJECTIVE: The aim of this study was to evaluate the proliferation and odontogenic differentiation of human dental pulp cells (hDPCs) and human umbilical cord mesenchymal stem cells (hUCMSCs) in three-dimensional co-culture system which was established with the help of bone morphogenetic protein-2 (BMP-2) and hydrogel. METHODS: hDPCs and hUCMSCs were cultured in different concentrations of hydrogel to explore the more suitable concentrations for subsequent experiments. hUCMSCs and hDPCs induced by BMP-2 were co-cultured in the hydrogel. MTT assay was used to measure the cell viability. The differentiation into odontoblast-like cells were measured by the mRNA expression of dentin salivary phosphoprotein (DSPP), dentin matrix protein-1 (DMP-1), alkaline phosphatase and osteocalcin. Alizarin red staining was performed for the formation of mineralized nodules. RESULTS: hUCMSCs and hDPCs could grow and proliferate in hydrogel scaffold. The growth rate of cells in lower concentrations hydrogels were higher than that of high concentrations hydrogels (P < 0.05). The study showed that 0.25% hydrogel scaffold was more suitable for subsequent experiments than other groups. Compared with hUCMSCs-monoculture and hDPCs-monoculture, the co-culture groups exhibited more proliferative potential, alkaline phosphatase activity and mineralization nodule formation (P < 0.05). The mRNA expression in co-culture groups were higher than that of hUCMSCs-monoculture, closed to or even higher than that of hDPCs-monoculture. CONCLUSION: 0.25% hydrogel was the suitable concentration in co-culture system for subsequent experiments. The co-culture groups had stronger abilities of odontoblastic differentiation and mineralization than cells-monoculture groups, indicated that the co-culture conditions could regulate cell proliferation and differentiation within a certain range.


Assuntos
Diferenciação Celular , Polpa Dentária/citologia , Hidrogéis , Células-Tronco Mesenquimais/citologia , Fosfatase Alcalina/genética , Proliferação de Células , Células Cultivadas , Técnicas de Cocultura , Proteínas da Matriz Extracelular/genética , Humanos , Odontoblastos/citologia , Osteocalcina/genética , Fosfoproteínas/genética , Sialoglicoproteínas/genética , Cordão Umbilical/citologia
11.
Arch Oral Biol ; 109: 104584, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31630006

RESUMO

OBJECTIVES: To investigate whether rutin could protect human periodontal ligament stem cells (hPDLSCs) from TNF-α induced damage to osteogenic differentiation in inflammatory environment and detect the underlying mechanism. MATERIALS AND METHODS: hPDLSCs were identified by flow cytometery. TNF-α was used to stimulate hPDLSCs to establish an inflammation model in vitro. Alkaline phosphatase (ALP) staining, ALP activity test, and Alizarin Red staining were used to detect the changes of osteogenic differentiation ability. The mRNA and protein levels of osteogenic genes were evaluated by RT-PCR and Western Blot. The expression of mTOR was also detected by Western Blot. RESULTS: hPDLSCs were positive to MSCs specific surface markers. The inflammatory environment in vitro could be established by stimulating hPDLSCs with TNF-α (20 ng/mL). TNF-α (20 ng/mL) could decrease the ALP activity and mineralization ability of hPDLSCs and down-regulate the expression of osteogenic genes in inflammatory environment. Moreover, rutin could affect TNF-α (20 ng/mL) induced damage to osteogenic differentiation of hPDLSCs in a dose-dependent manner, 10 µmol/L rutin could significantly reverse the damage caused by TNF-α. In addition, rutin inhibited TNF-α-activated mTOR signal transduction by inhibiting the phosphorylation of mTOR, similar to the effects of rapamycin(a specific mTOR inhibitor). CONCLUSIONS: Rutin could protect hPDLSCs from TNF-α induced damage to osteogenic differentiation in inflammatory environment, and rutin is expected to become a new candidate drug for the treatment of bone defect of periodontitis.


Assuntos
Osteogênese , Ligamento Periodontal/citologia , Rutina/farmacologia , Transdução de Sinais , Células-Tronco/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo , Fator de Necrose Tumoral alfa/efeitos adversos , Fosfatase Alcalina/metabolismo , Diferenciação Celular , Células Cultivadas , Humanos , Inflamação , Células-Tronco/citologia
12.
Chemosphere ; 239: 124668, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31494325

RESUMO

Nitrite is one of major environmental pollutants that can impact immunological parameters in aquatic organisms. In the present study, we investigated the effects of nitrite exposure on oxidative stress, DNA damage and apoptosis in mud crab (Scylla paramamosain). Mud crab were exposed to 0, 5, 10 and 15 mg L-1 nitrite for 72 h. These data showed that acid phosphatase (ACP) and alkaline phosphatase (ALP) activity significantly decreased in treatments with various concentrations of nitrite (5, 10 and 15 mg L-1) after 24 and 48 h, while the levels of nitric oxide (NO) significantly increased in these treatments. Nitrite exposure could suppress superoxide dismutase (SOD) and catalase (CAT) activity, and increase the formation of malondialdehyde (MDA) after 48 and 72 h of exposure. In addition, nitrite exposure decreased total haemocyte counts after 48 and 72 h of exposure. Cytological damage, DNA damage and apoptosis was observed obviously at 72 h after nitrite exposure. Moreover, nitrite exposure significantly induced the mRNA levels of phosphorylated Jun N-terminal kinases (JNK), and eventually activated p53 signaling and caspase-3. These results indicated that nitrite exposure could induce oxidative stress, which further caused DNA damage and apoptosis in mud crab. Our results will be helpful to understand the mechanism of nitrite toxicity on crustaceans.


Assuntos
Apoptose/efeitos dos fármacos , Braquiúros/efeitos dos fármacos , Dano ao DNA/genética , Nitritos/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Fosfatase Ácida/metabolismo , Fosfatase Alcalina/metabolismo , Animais , Braquiúros/genética , Catalase/metabolismo , Hemócitos , Malondialdeído/metabolismo , Óxido Nítrico/metabolismo , Transdução de Sinais , Superóxido Dismutase/metabolismo
13.
Int Endod J ; 53(1): 72-83, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31419325

RESUMO

AIM: To comparatively evaluate changes in the proliferation and mineralization abilities of dental pulp stem cells (DPSCs) from juvenile and adult rats in a lipopolysaccharide (LPS)-induced inflammatory microenvironment to provide a theoretical basis for the age-related differences observed in DPSCs during repair of inflammatory injuries. METHODOLOGY: DPSCs were isolated from juvenile (JDPSCs) and adult rats (ADPSCs), and senescence-associated ß-galactosidase staining was used to compare senescence between JDPSCs and ADPSCs. Effects of LPS on JDPSCs and ADPSCs proliferation were investigated by cell counting kit-8 assays and flow cytometry. Alizarin red staining, quantitative reverse transcription polymerase chain reaction and Western blot assay were used to examine the effects of LPS on mineralization-related genes and proteins in JDPSCs and ADPSCs. Immunohistochemistry was used to compare interleukin-1ß (IL-1ß) and osteocalcin (OCN) expression in the pulpitis model. Unpaired Student's t-tests and one-way anova were used for statistical analysis. RESULTS: DPSCs were isolated from juvenile and adult rat dental pulp tissues. At low concentrations (0.1-1 µg mL-1 ), LPS significantly promoted the proliferation of JDPSCs (P < 0.01) and ADPSCs (P < 0.01 or P < 0.05), with the effect being stronger in JDPSCs than in ADPSCs. In addition, mineralized nodules and the expression of mineralization-related genes (OCN, DSPP, ALP, BSP) increased significantly after stimulation with LPS (0.5 µg mL-1 ) in JDPSCs and ADPSCs (P < 0.01 or P < 0.05), and JDPSCs displayed a more obvious increase than ADPSCs. Western blots revealed OCN and ALP expression levels in JDPSCs treated with LPS were significantly upregulated (P < 0.05); meanwhile, ALP expression in ADPSCs increased slightly but significantly (P < 0.05), and OCN expression was not affected. Finally, IL-1ß expression was significantly higher (P < 0.05) and OCN expression was significantly lower (P < 0.05) in the inflamed dental pulp of adult rats than in juvenile rats. CONCLUSIONS: A certain degree of inflammatory stimulation promoted the proliferation and mineralization of DPSCs; however, this effect declined with age. The DPSCs of adult donors in an inflammatory microenvironment have a weaker repair ability than that of juvenile donors, who are better candidates for tissues damage repair.


Assuntos
Polpa Dentária , Células-Tronco , Afeto , Fosfatase Alcalina , Animais , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Humanos , Ratos
14.
Shanghai Kou Qiang Yi Xue ; 28(4): 356-361, 2019 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-31792473

RESUMO

PURPOSE: To explore the effect of rutin on osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment. METHODS: Periodontal ligament stem cells (PDLSCs) were obtained by limited dilution method in vitro. PDLSCs were identified by flow cytometery. Lipopolysaccharide(LPS) was used to stimulate human periodontal ligament stem cells to establish an inflammation model in vitro. The experiment was divided into 4 groups: in group 1, only α-MEM was used to culture PDLSCs; in group 2, α-MEM medium containing LPS was used to culture PDLSCs, in group 3, rutin was added to α-MEM medium containing LPS to PDLSCs; and in group 4, α-MEM medium containing rutin was used to culture PDLSCs. Cell counting kit-8 was used to detect cell proliferation activity. Alkaline phosphatase(ALP) staining, ALP activity test, alizarin red staining, RT-PCR, and Western blot were used to detect the changes of osteogenic differentiation ability. The data were analyzed by SPSS 17.0 software package. RESULTS: The results of CCK-8 and ALP activity analysis showed that rutin at 10 µmol/L could significantly promote the proliferation and differentiation of periodontal stem cells under inflammatory state. ALP staining and alizarin red staining proved that (10 µmol/L) rutin could improve osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment. RT-PCR and Western blot results showed that rutin could enhance the expression of osteogenic genes and proteins such as COL1, ALP, and RUNX2 under inflammatory state. CONCLUSIONS: Rutin can promote osteogenic differentiation of periodontal ligament stem cells under inflammatory microenvironment.


Assuntos
Osteogênese , Ligamento Periodontal , Fosfatase Alcalina , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Humanos , Rutina , Células-Tronco
15.
J Environ Sci (China) ; 86: 38-49, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31787189

RESUMO

Three fractions of alkaline phosphatase activity (APA), including phytoplankton APA (phyto-APA), bacterial APA (bact-APA), and free-APA, were examined in the sea surface microlayer (SML) and the subsurface water (SSW) from Daya Bay, Guishan Island, and Guanghai Bay of southern China. Relationships between APA and environmental parameters were analyzed. The growth of phytoplankton was significantly limited by dissolved inorganic phosphorus (DIP) in the three sea areas, especially in Daya Bay. Total-APA ranged between 1.41 and 35.26 nmol/L/hr, and the highest value was found in Daya Bay. The increased APA in Daya Bay was the result of the increase of phytoplankton biomass and the response of phytoplankton to P limitation. Phyto-APA was the main contributor in Daya Bay, while phyto- and free-APA co-dominated in Guishan Island and Guanghai Bay. Bact-, phyto-, and total-APA showed a significant inverse power function relationship with DIP, and 0.2 µmol/L was the threshold for DIP on particulates and total-APA. Pearson correlation analysis suggested that DIP limitation together with high N levels enhanced APA. High water temperature and freshwater input accelerated APA as well. Principal component analysis clearly separated samples from the three sea areas, as well as from the SML and the SSW, which indicated the differences in environmental parameters and APA levels. Our results highlight the influence of phosphorus limitation and environmental parameters on APA.


Assuntos
Fosfatase Alcalina/análise , Monitoramento Ambiental , Fósforo/análise , Fitoplâncton/crescimento & desenvolvimento , Biomassa , China
16.
Chem Commun (Camb) ; 55(98): 14852-14855, 2019 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-31769450

RESUMO

A self-delivery system consisting of lonidamine and a self-assembling peptide was designed for the selective killing of phosphatase-overexpressing cancer cells, which was mediated by both enhanced cellular uptake of LND-peptide and enzyme-triggered intracellular fiber formation, thereby providing a generalized strategy to develop cancer-targeting systems of drug conjugates.


Assuntos
Fosfatase Alcalina/metabolismo , Antineoplásicos/química , Indazóis/química , Peptídeos/química , Fosfatase Alcalina/genética , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Hidrogéis/química , Camundongos , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Transplante Heterólogo
17.
Zhonghua Er Ke Za Zhi ; 57(11): 899-903, 2019 Nov 02.
Artigo em Chinês | MEDLINE | ID: mdl-31665851
18.
Int J Nanomedicine ; 14: 8271-8284, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31686820

RESUMO

Background: Salvianolic acid B has been proven as an effective drug to promote osteogenesis and angiogenesis which could be beneficial for bone repair. Purpose: The objective of this study was to construct a salvianolic acid B-loaded chitosan/hydroxyapatite (Sal B-CS/HA) bone scaffold with controlled release and effective bioactivity. Methods: The characteristics, controlled release behavior and bioactivity of Sal B-CS/HA scaffold were evaluated in vitro. The bone repair effect was evaluated in the rabbit radius defect model. Results: The results showed that chemical and physical characteristics of salvianolic acid B and chitosan/hydroxyapatite (CS/HA) material did not obviously change after the drug loading procedure; the drug release of salvianolic acid B was stable and continuous from the Sal B-CS/HA scaffold for 8 weeks in vitro; the biocompatibility of the Sal B-CS/HA was favorable by evaluation of cell morphology and proliferation; the osteogenic and angiogenic bioactivities of the Sal B-CS/HA scaffold were proved to be effective by in vivo and in vitro tests. Conclusion: Our results suggest that this salvianolic acid B-loaded bone scaffold has potential to be used for bone defect repair with both osteogenic and angiogenic bioactivities.


Assuntos
Benzofuranos/farmacologia , Osso e Ossos/irrigação sanguínea , Osso e Ossos/patologia , Quitosana/química , Durapatita/química , Neovascularização Fisiológica/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Tecidos Suporte/química , Fosfatase Alcalina/metabolismo , Animais , Osso e Ossos/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Liberação Controlada de Fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Camundongos , Coelhos , Espectroscopia de Infravermelho com Transformada de Fourier , Tomografia Computadorizada por Raios X , Fator A de Crescimento do Endotélio Vascular/metabolismo
19.
Comput Biol Chem ; 83: 107142, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31698161

RESUMO

Phosphorus is one of the primary macronutrient of plants, which is present in soil. It is essential for normal growth and development of plants. Plants use inorganic form of phosphate but organic form can also be assimilated with the help of soil inhabiting bacteria. Alkaline phosphatase is an enzyme present in Rizobium bacteria. This enzyme is responsible for solubilization and mineralization of organic phosphate and makes it readily available for plants. In the present study, nine different strains of Rhizobium leguminosarum were selected for a detailed computational structural and functional characterization and phylogenetic studies of alkaline phosphatase. Amino acid sequences were retrieved from UniProt and saved in FASTA format for use in analysis. Phylogenetic analysis of these strains was done by using MEGA7. 3D structure prediction was performed by using online server I-Tasser. Galaxy Web and 3D Refine were used for structure refinement. The refined structures were evaluated using two validation servers, QMEAN and SAVES. Protein-protein interaction analysis was done by using STRING. For detailed functional characterization, Cofactor, Coach, RaptorX, PSORT and MEME were used. Overall quality of predicted protein models was above 80%. Refined and validated models were submitted into PMDB. Seven out of nine strains were closely related and other two were distantly related. Protein-Protein interaction showed no significant co-expression among the interaction partners.


Assuntos
Fosfatase Alcalina/química , Fosfatase Alcalina/metabolismo , Simulação por Computador , Filogenia , Rhizobium leguminosarum/enzimologia , Sequência de Aminoácidos , Modelos Moleculares , Ligação Proteica , Conformação Proteica
20.
Mol Cell Biochem ; 462(1-2): 173-183, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31620952

RESUMO

Osteoblasts and osteoclasts play essential and opposite roles in maintaining bone homeostasis. Osteoblasts fill cavities excavated by osteoclasts. The mevalonate pathway provides essential prenyl pyrophosphates for the activities of GTPases that promote differentiation of osteoclasts but suppress that of osteoblasts. Preclinical and clinical studies suggest that mevalonate suppressors such as statins increase bone mineral density and reduce risk of bone fracture. Tocotrienols down-regulate 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, the rate-limiting enzyme in the mevalonate pathway. In vivo studies have shown the bone-protective activity of tocotrienols. We hypothesize that d-δ-tocotrienol, a mevalonate suppressor, induces differentiation of murine MC3T3-E1 preosteoblasts. Alizarin staining showed that d-δ-tocotrienol (0-25 µmol/L) induced mineralized nodule formation in a concentration-dependent manner in MC3T3-E1 preosteoblasts. d-δ-Tocotrienol (0-25 µmol/L), but not D-α-tocopherol (25 µmol/L), significantly induced alkaline phosphatase activity, an indicator of preosteoblast differentiation. The expression of differentiation marker genes including BMP-2 and VEGFα was stimulated dose dependently by d-δ-tocotrienol (0-25 µmol/L). Concomitantly, Western blot analysis showed that d-δ-tocotrienol down-regulated HMG CoA reductase. d-δ-Tocotrienol (0-25 µmol/L) had no impact on the viability of MC3T3-E1 preosteoblasts following 48-h incubation, suggesting lack of cytotoxicity at these doses. Tocotrienols and other mevalonate suppressors have potential in maintaining bone health.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Osteoblastos/citologia , Vitamina E/análogos & derivados , Fosfatase Alcalina/metabolismo , Animais , Linhagem Celular , Regulação para Baixo/efeitos dos fármacos , Hidroximetilglutaril-CoA Redutases/metabolismo , Ácido Mevalônico/metabolismo , Camundongos , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteogênese/efeitos dos fármacos , Vitamina E/farmacologia , Proteínas ras/metabolismo
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