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1.
Int Dent J ; 73(3): 435-442, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36402577

RESUMO

OBJECTIVES: The COVID-19 vaccine is currently being administered worldwide to address the ongoing pandemic. Although these vaccines have proven effective in preventing severe disease, the level of immunity required to prevent respiratory mucosal infection remains less well understood. Therefore, it is desirable to develop a noninvasive screening strategy such as oral fluid to monitor secreted antibodies longitudinally as potential surrogates of mucosal immunity. METHODS: We evaluated the anti-spike protein antibodies in gingival crevicular fluid (GCF) and saliva and compared them to immune responses in the blood of 50 healthy health care workers following 2 doses of intramuscular Pfizer/BioNTech-BNT162b2 vaccine. RESULTS: The antibodies to SARS-CoV-2 spike and subdomain proteins (RBD, S1, S2, and NTD) were significantly higher in serum than oral fluids but showed a greater detection rate and higher median titres in GCF than saliva. For all tested SARS-CoV-2 antigens, IgG in GCF (as opposed to saliva) showed a more significant and stronger correlation with IgG in serum. Serum-neutralising antibodies (Nab) titres also displayed a significant and stronger correlation with anti-spike protein and their subdomains in GCF than saliva. Interestingly, the time post-second dose of vaccine and sex had a similar influence on IgG in serum and GCF. However, interferon (IFN)-γ-producing T-cell responses showed no association with SARS-Cov-2 IgG antibodies in serum, GCF, or saliva and neutralisation antibodies in serum. The correlation matrix of all measured parameters grouped serum and GCF IgG parameters separately from salivary IgG parameters indicating that GCF better represents the humoural response in serum than saliva. CONCLUSIONS: Within limitations, we propose that GCF could be a less invasive alternative to serum and more appropriate than saliva to detect antibody responses by current COVID-19 vaccines if the GCF collection procedure could be standardised. Further research is needed to investigate the suitability of GCF for community immune surveillance for vaccines.


Assuntos
Vacina BNT162 , COVID-19 , Humanos , Vacinas contra COVID-19 , COVID-19/prevenção & controle , SARS-CoV-2 , Imunoglobulina G , Imunidade , Anticorpos Antivirais
2.
Int J Mol Sci ; 22(23)2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34884821

RESUMO

Reconstruction of bone due to surgical removal or disease-related bony defects is a clinical challenge. It is known that the immune system exerts positive immunomodulatory effects on tissue repair and regeneration. In this study, we evaluated the in vivo efficacy of autologous neutrophils on bone regeneration using a rabbit calvarial defect model. Methods: Twelve rabbits, each with two surgically created calvarial bone defects (10 mm diameter), were randomly divided into two groups; (i) single application of neutrophils (SA-NP) vs. SA-NP control, and (ii) repetitive application of neutrophils (RA-NP) vs. RA-NP control. The animals were euthanized at 4 and 8 weeks post-operatively and the treatment outcomes were evaluated by micro-computed tomography, histology, and histomorphometric analyses. Results: The micro-CT analysis showed a significantly higher bone volume fraction (bone volume/total volume) in the neutrophil-treated groups, i.e., median interquartile range (IQR) SA-NP (18) and RA-NP (24), compared with the untreated controls, i.e., SA-NP (7) and RA-NP (14) at 4 weeks (p < 0.05). Similarly, new bone area fraction (bone area/total area) was significantly higher in neutrophil-treated groups at 4 weeks (p < 0.05). Both SA-NP and RA-NP had a considerably higher bone volume and bone area at 8 weeks, although the difference was not statistically significant. In addition, immunohistochemical analysis at 8 weeks revealed a higher expression of osteocalcin in both SA-NP and RA-NP groups. Conclusions: The present study provides first hand evidence that autologous neutrophils may have a positive effect on promoting new bone formation. Future studies should be performed with a larger sample size in non-human primate models. If proven feasible, this new promising strategy could bring clinical benefits for bone defects to the field of oral and maxillofacial surgery.


Assuntos
Regeneração Óssea , Neutrófilos/metabolismo , Crânio/fisiologia , Animais , Doenças Ósseas/terapia , Modelos Animais de Doenças , Masculino , Neutrófilos/transplante , Osteocalcina/metabolismo , Coelhos , Crânio/diagnóstico por imagem , Crânio/patologia , Microtomografia por Raio-X
4.
Heliyon ; 5(12): e02900, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31890935

RESUMO

BACKGROUND: Awareness of feeding among mothers plays an important role in the growth and development of children with cleft lip and palate. Mothers' increased awareness provides the right care and nutrition to these children in order to grow and achieve their developmental milestones. OBJECTIVES: This study aimed at (1) assessing mothers' awareness on breastfeeding and formula-milk feeding, weaning, growth, and development of infants with cleft lip and palate and (2) the relationship of mothers' awareness with their level of education and monthly household income. METHODS: This cross-sectional pilot study included all the mothers of infants aged up to one year who had cleft lip and/or palate attended Dental Teaching Hospital, Peradeniya between the years 2015 and 2016. Mothers of infants with oral clefts associated with other syndromic features and chronic illnesses were excluded. A pre-tested interviewer-administered questionnaire was designed to collect data. To assess the clarity of the questions, prior to the study, the questionnaire was pre-tested in 10 mothers of infants with an oral cleft but not selected for the study. Frequency distributions and descriptive statistics were used to describe study variables. Spearman Rank test was used to determine the association of mothers' awareness with their level of education and monthly household income with a 95% confidence interval. RESULTS: The sample consisted of 101 mothers who had infants born with a cleft and the response rate was 100%. Except one, all mothers reported that they have received overall feeding instructions either from a doctor, nurse, cleft center staff or a midwife. Over 65.3% of mothers were aware of feeding-related factors including; breastfeeding, formula-milk feeding, and weaning of infants with a cleft. Overall more than 80% of mothers were aware of factors related to growth and development of the child including urination, sleeping pattern, weight and growth charts. The education level of mothers was not significantly associated with mothers' awareness of any factors related to breastfeeding, formula-milk feeding, weaning and growth, and development. Further, mothers' awareness of breastmilk as the best food for babies under 6 months (p = 0.028), weaning at 4-6 months (p = 0.024), replacing milk feeds by weaning foods (p = 0.02) and not providing junk foods to infants in between main meals (p = 0.01) were significantly associated with monthly household income. CONCLUSIONS: The findings of this study suggest that mothers were aware of factors related to feeding infants with a cleft lip and palate and their growth and development. The awareness of mothers about certain factors related to feeding and weaning was significantly associated with monthly household income.

5.
Nutrients ; 10(9)2018 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-30181524

RESUMO

Human milk is nutrient rich, complex in its composition, and is key to a baby's health through its role in nutrition, gastrointestinal tract and immune development. Seventy-eight mothers (19⁻42 years of age) of Asian, Maori, Pacific Island, or of European ethnicity living in Manawatu-Wanganui, New Zealand (NZ) completed the study. The women provided three breast milk samples over a one-week period (6⁻8 weeks postpartum), completed a three-day food diary and provided information regarding their pregnancy and lactation experiences. The breast milk samples were analyzed for protein, fat, fatty acid profile, ash, selected minerals (calcium, magnesium, selenium, zinc), and carbohydrates. Breast milk nutrient profiles showed no significant differences between the mothers of different ethnicities in their macronutrient (protein, fat, carbohydrate, and moisture) content. The breast milk of Asian mothers contained significantly higher levels of polyunsaturated fatty acids (PUFAs), omega-3 (n-3) and omega-6 (n-6) fatty acids, docosahexaenoic acid (DHA), and linoleic acids. Arachidonic acid was significantly lower in the breast milk of Maori and Pacific Island women. Dietary intakes of protein, total energy, saturated and polyunsaturated fat, calcium, phosphorus, zinc, iodine, vitamin A equivalents, and folate differed between the ethnic groups, as well as the number of serves of dairy foods, chicken, and legumes. No strong correlations between dietary nutrients and breast milk components were found.


Assuntos
Dieta , Etnicidade , Leite Humano/química , Adulto , Ácido Araquidônico/análise , Aleitamento Materno , Registros de Dieta , Gorduras na Dieta/análise , Fibras na Dieta/análise , Proteínas Alimentares/análise , Ácidos Docosa-Hexaenoicos/análise , Ácido Eicosapentaenoico/análise , Ácidos Graxos Ômega-6/análise , Feminino , Humanos , Lactação , Micronutrientes/análise , Nova Zelândia , Avaliação Nutricional , Ilhas do Pacífico , Período Pós-Parto , Inquéritos e Questionários , Adulto Jovem
6.
J Tissue Eng Regen Med ; 12(2): e1221-e1236, 2018 02.
Artigo em Inglês | MEDLINE | ID: mdl-28715156

RESUMO

Repair and regeneration of critical-sized bone defects remain a major challenge in orthopaedic and craniomaxillofacial surgery. Until now, attempts to bioengineer bone tissue have been hindered by the inability to establish proper angiogenesis and osteogenesis in the tissue construct. In the present study, we established a novel triple cell co-culture model consisting of osteoblasts, endothelial cells, and neutrophils and conducted a systematic investigation of the effects of neutrophils on angiogenesis and osteogenesis. Neutrophils significantly increased angiogenesis in the tissue construct, evidenced by the formation of microvessel-like structures with an extensive lattice-like, stable tubular network in the co-culture model. Moreover, neutrophils significantly induced the expression of pro-angiogenic markers, such as VEGF-A, CD34, EGF, and FGF-2 in a dose- and time-dependent manner. Subsequently, PCR arrays corroborated that neutrophils upregulate the important angiogenic markers and MMPs. Moreover, neutrophils also enhanced osteogenic markers, such as ALP, OCN, OPN, and COL-1 compared with the controls. As shown by the osteogenic gene arrays, neutrophils significantly regulated major osteogenic markers such as BMP2, BMP3, BMP4, BMP5, TGF-ß2, RUNX2, and ECM proteins. Significantly higher mineralization was observed in triple cell co-culture compared with controls. Foregoing data indicate that the triple cell co-culture model can be used to stimulate the growth of microvasculature within a bone bioengineering construct to improve cell viability. Neutrophil-mediated enhancement of angiogenesis and osteogenesis could be a viable, clinically relevant tissue engineering strategy to obtain optimal bone growth in defect sites, in the field of oral and maxillofacial surgery.


Assuntos
Técnicas de Cocultura/métodos , Células Endoteliais da Veia Umbilical Humana/citologia , Neovascularização Fisiológica , Neutrófilos/citologia , Osteoblastos/citologia , Osteogênese , Adulto , Biomarcadores/metabolismo , Matriz Óssea/metabolismo , Calcificação Fisiológica , Forma Celular , Feminino , Regulação da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Neovascularização Fisiológica/genética , Neutrófilos/metabolismo , Osteogênese/genética , Fatores de Tempo , Migração Transendotelial e Transepitelial , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto Jovem
7.
Sci Rep ; 6: 29829, 2016 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-27538450

RESUMO

Periodontal (gum) disease is a highly prevalent infection and inflammation accounting for the majority of tooth loss in adult population worldwide. Porphyromonas gingivalis is a keystone periodontal pathogen and its lipopolysaccharide (PgLPS) acts as a major virulence attribute to the disease. Herein, we deciphered the overall host response of human gingival fibroblasts (HGFs) to two featured isoforms of tetra-acylated PgLPS1435/1449 and penta-acylated PgLPS1690 with reference to E. coli LPS through quantitative proteomics. This study unraveled differentially expressed novel biomarkers of immuno-inflammatory response, antioxidant defense and cytoskeletal dynamics in HGFs. PgLPS1690 greatly upregulated inflammatory proteins (e.g. cyclophilin, inducible nitric oxide synthase, annexins, galectin, cathepsins and heat shock proteins), whereas the anti-inflammatory proteins (e.g. Annexin A2 and Annexin A6) were significantly upregulated by PgLPS1435/1449. Interestingly, the antioxidants proteins such as mitochondrial manganese-containing superoxide dismutase and peroxiredoxin 5 were only upregulated by PgLPS1690. The cytoskeletal rearrangement-related proteins like myosin were differentially regulated by these PgLPS isoforms. The present study gives new insight into the biological properties of P. gingivalis LPS lipid A moiety that could critically modulate immuno-inflammatory response, antioxidant defense and cytoskeletal dynamics in HGFs, and thereby enhances our understanding of periodontal pathogenesis.


Assuntos
Antioxidantes/metabolismo , Fibroblastos/metabolismo , Gengiva/metabolismo , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos/toxicidade , Porphyromonas gingivalis/química , Células Cultivadas , Fibroblastos/patologia , Gengiva/patologia , Humanos , Inflamação/metabolismo , Inflamação/patologia , Lipopolissacarídeos/química
8.
Food Funct ; 7(4): 1825-32, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26778667

RESUMO

The fate of stable-isotope (13)C labelled and non-labelled inulin catabolism by the gut microbiota was assessed in a healthy rat model. Sprague-Dawley male rats were randomly assigned to diets containing either cellulose or inulin, and were fed these diets for 3 days. On day (d) 4, rats allocated to the inulin diet received (13)C-labelled inulin. The rats were then fed the respective non-labelled diets (cellulose or inulin) until sampling (d4, d5, d6, d7, d10 and d11). Post feeding of (13)C-labelled substrate, breath analysis showed that (13)C-inulin cleared from the host within a period of 36 hours. Faecal (13)C demonstrated the clearance of inulin from gut with a (13)C excess reaching maximum at 24 hours (d5) and then declining gradually. There were greater variations in caecal organic acid concentrations from d4 to d6, with higher concentrations of acetic, butyric and propionic acids observed in the rats fed inulin compared to those fed cellulose. Inulin influenced caecal microbial glycosidase activity, increased colon crypt depth, and decreased the faecal output and polysaccharide content compared to the cellulose diet. In summary, the presence of inulin in the diet positively influenced large bowel microbial fermentation.


Assuntos
Bactérias/metabolismo , Ceco/metabolismo , Intestino Grosso/metabolismo , Inulina/metabolismo , Animais , Isótopos de Carbono/química , Isótopos de Carbono/metabolismo , Ceco/microbiologia , Carboidratos da Dieta/análise , Carboidratos da Dieta/metabolismo , Fezes/química , Fermentação , Microbioma Gastrointestinal , Intestino Grosso/microbiologia , Inulina/química , Marcação por Isótopo , Masculino , Ratos , Ratos Sprague-Dawley
9.
Nutr Res ; 35(1): 23-34, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25530011

RESUMO

This study examined the effect of a Boysenberry beverage (750 mg polyphenols), an apple fiber beverage (7.5 g dietary fiber), and a Boysenberry plus apple fiber beverage (750 mg polyphenols plus 7.5 g dietary fiber) on gut health. Twenty-five individuals completed the study. The study was a placebo-controlled crossover study, where every individual consumed 1 of the 4 treatments in turn. Each treatment phase was 4-week long and was followed by a 2-week washout period. The trial beverages were 350 g taken in 2 doses every day (ie, 175 mL taken twice daily). The hypothesis for the study was that the combination of polyphenols and fiber would have a greater benefit on gut health than the placebo product or the fiber or polyphenols on their own. There were no differences in fecal levels of total bacteria, Bacteroides-Prevotella-Porphyromonas group, Bifidobacteriumspecies, Clostridium perfringens, or Lactobacillus species among any of the treatment groups. Fecal short chain fatty acid concentrations did not vary among treatment groups, although prostaglandin E2 concentrations were higher after consumption of the Boysenberry juice beverage. No significant differences were found in quantitative measures of gut health between the Boysenberry juice beverage, the apple fiber beverage, the Boysenberry juice plus apple fiber beverage, and the placebo beverage.


Assuntos
Bebidas/análise , Fibras na Dieta/administração & dosagem , Ácidos Graxos Voláteis/análise , Fezes/microbiologia , Frutas/química , Polifenóis/administração & dosagem , Adulto , Bacteroides/isolamento & purificação , Bifidobacterium/isolamento & purificação , Índice de Massa Corporal , Peso Corporal , Clostridium perfringens/isolamento & purificação , Estudos Cross-Over , DNA Bacteriano/isolamento & purificação , Dinoprostona/análise , Fezes/química , Feminino , Humanos , Imunoglobulina A/análise , Lactobacillus/isolamento & purificação , Masculino , Malus/química , Pessoa de Meia-Idade , Cooperação do Paciente , Porphyromonas/isolamento & purificação , Prevotella/isolamento & purificação
10.
Mol Nutr Food Res ; 58(12): 2350-7, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25176606

RESUMO

SCOPE: Selenium (Se) is a micronutrient essential for human health, including immune function. Previous research indicates that Se supplementation may cause a shift from T helper (Th)1- to Th2-type immune responses. We aim to test the potential health promoting effects of Se-enriched broccoli. METHODS AND RESULTS: In a human trial, 18 participants consumed control broccoli daily for 3 days. After a 3-day wash-out period, the participants were provided with Se-enriched broccoli containing 200 µg of Se per serving for 3 days. Plasma and peripheral blood mononuclear cell (PBMC) samples were collected at the start and end of each broccoli feeding period for analysis of total Se and measurement of cytokine production from PBMC stimulated with antigens ex vivo. Plasma Se content remained consistent throughout the control broccoli feeding period and the baseline of the Se-enriched broccoli period (1.22 µmol/L) and then significantly increased following 3 days of Se-enriched broccoli feeding. Interleukin (IL-2, IL-4, IL-5, IL-13, and IL-22) production from PBMC significantly increased after 3 days of Se-enriched broccoli feeding compared with baseline. CONCLUSION: This study indicates that consumption of Se-enriched broccoli may increase immune responses toward a range of immune challenges.


Assuntos
Brassica/química , Leucócitos Mononucleares/efeitos dos fármacos , Selênio/administração & dosagem , Adulto , Idoso , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Glucosinolatos/urina , Humanos , Interleucina-13/metabolismo , Interleucina-2/metabolismo , Interleucina-4/metabolismo , Interleucina-5/metabolismo , Interleucinas/metabolismo , Masculino , Pessoa de Meia-Idade , Selênio/sangue , Selenoproteína P/sangue , Adulto Jovem , Interleucina 22
11.
PLoS One ; 8(3): e58496, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23554896

RESUMO

BACKGROUND: Porphyromonas gingivalis is a major pathogen of periodontal disease that affects a majority of adults worldwide. Increasing evidence shows that periodontal disease is linked to various systemic diseases like diabetes and cardiovascular disease, by contributing to increased systemic levels of inflammation. Lipopolysaccharides (LPS), as a key virulent attribute of P. gingivalis, possesses significant amount of lipid A heterogeneity containing tetra- (LPS1435/1449) and penta-acylated (LPS1690) structures. Hitherto, the exact molecular mechanism of P. gingivalis LPS involved in periodontal pathogenesis remains unclear, due to limited understanding of the specific receptors and signaling pathways involved in LPS-host cell interactions. METHODOLOGY/PRINCIPAL FINDINGS: This study systematically investigated the effects of P. gingivalis LPS1435/1449 and LPS1690 on the expression of TLR2 and TLR4 signal transduction and the activation of pro-inflammatory cytokines IL-6 and IL-8 in human gingival fibroblasts (HGFs). We found that LPS1435/1449 and LPS1690 differentially modulated TLR2 and TLR4 expression. NF-κB pathway was significantly activated by LPS1690 but not by LPS1435/1449. In addition, LPS1690 induced significant expression of NF-κB and p38 MPAK pathways-related genes, such as NFKBIA, NFKB1, IKBKB, MAP2K4 and MAPK8. Notably, the pro-inflammatory genes including GM-CSF, CXCL10, G-CSF, IL-6, IL-8 and CCL2 were significantly upregulated by LPS1690 while down-regulated by LPS1435/1449. Blocking assays confirmed that TLR4-mediated NF-κB signaling was vital in LPS1690-induced expression of IL-6 and IL-8 in HGFs. CONCLUSIONS/SIGNIFICANCE: The present study suggests that the tetra- and penta-acylated lipid A structures of P. gingivalis LPS differentially activate TLR4-mediated NF-κB signaling pathway, and significantly modulate the expression of IL-6 and IL-8 in HGFs. The ability to alter the lipid A structure of LPS could be one of the strategies carried-out by P. gingivalis to evade innate host defense in gingival tissues, thereby contributing to periodontal pathogenesis.


Assuntos
MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Gengiva/metabolismo , Lipídeo A/farmacologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , NF-kappa B/metabolismo , Porphyromonas gingivalis/química , Receptor 4 Toll-Like/metabolismo , Acilação , Citocinas/biossíntese , Citocinas/imunologia , MAP Quinases Reguladas por Sinal Extracelular/genética , Fibroblastos , Gengiva/imunologia , Gengiva/microbiologia , Gengiva/patologia , Humanos , Lipídeo A/química , Lipídeo A/imunologia , Sistema de Sinalização das MAP Quinases/imunologia , NF-kappa B/imunologia , Porphyromonas gingivalis/imunologia , Receptor 4 Toll-Like/imunologia
12.
BMC Microbiol ; 13: 73, 2013 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-23548063

RESUMO

BACKGROUND: Porphyromonas gingivalis lipopolysaccharide (LPS) is a crucial virulence factor strongly associated with chronic periodontitis which is the primary cause of tooth loss in adults. It exhibits remarkable heterogeneity containing tetra-(LPS(1435/1449)) and penta-(LPS(1690)) acylated lipid A structures. Human gingival fibroblasts (HGFs) as the main resident cells of human gingiva play a key role in regulating matrix metalloproteinases (MMPs) and contribute to periodontal homeostasis. This study investigated the expression and regulation of MMPs1-3 and tissue inhibitors of MMP-1 (TIMP-1) in HGFs in response to P. gingivalis LPS(1435/1449) and LPS(1690) and hexa-acylated E. coli LPS as a reference. The expression of MMPs 1-3 and TIMP-1 was evaluated by real-time PCR and ELISA. RESULTS: The MMP-3 mRNA and protein were highly upregulated in P. gingivalis LPS(1690)- and E. coli LPS-treated cells, whereas no induction was observed in P. gingivalis LPS(1435/1449)-treated cells. On the contrary, the expression of MMP-1 and -2 was not significantly affected by P. gingivalis LPS lipid A heterogeneity. The TIMP-1 mRNA was upregulated in P. gingivalis LPS(1435/1449)- and E. coli LPS-treated cells. Next, signal transduction pathways involved in P. gingivalis LPS-induced expression of MMP-3 were examined by blocking assays. Blockage of p38 MAPK and ERK significantly inhibited P. gingivalis LPS(1690)-induced MMP-3 expression in HGFs. CONCLUSION: The present findings suggest that the heterogeneous lipid A structures of P. gingivalis LPS differentially modulate the expression of MMP-3 in HGFs, which may play a role in periodontal pathogenesis.


Assuntos
Fibroblastos/metabolismo , Fibroblastos/microbiologia , Regulação da Expressão Gênica , Lipopolissacarídeos/toxicidade , Metaloproteinase 3 da Matriz/biossíntese , Porphyromonas gingivalis/patogenicidade , Adulto , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Perfilação da Expressão Gênica , Humanos , Lipopolissacarídeos/química , Metaloproteinase 1 da Matriz/biossíntese , Metaloproteinase 2 da Matriz/biossíntese , Reação em Cadeia da Polimerase em Tempo Real , Inibidor Tecidual de Metaloproteinase-1/biossíntese , Fatores de Virulência/química , Fatores de Virulência/toxicidade
13.
J Food Sci ; 77(10): H216-23, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22950602

RESUMO

UNLABELLED: The effects of red meat consumption with and without fermentable carbohydrates on indices of large bowel health in rats were examined. Sprague-Dawley rats were fed cellulose, potato fiber, or potato-resistant starch diets containing 12% casein for 2 wk, then similar diets containing 25% cooked beef for 6 wk. After week 8, cecal and colonic microbiota composition, fermentation end-products, colon structure, and colonocyte DNA damage were analyzed. Rats fed potato fiber had lower Bacteroides-Prevotella-Porphyromonas group compared to other diet groups. Colonic Bifidobacterium spp. and/or Lactobacillus spp. were higher in potato fiber and potato-resistant starch diets than in the cellulose diet. Beneficial changes were observed in short-chain fatty acid concentrations (acetic, butyric, and propionic acids) in rats fed potato fiber compared with rats fed cellulose. Phenol and p-cresol concentrations were lower in the cecum and colon of rats fed potato fiber. An increase in goblet cells per crypt and longer crypts were found in the colon of rats fed potato fiber and potato-resistant starch diets. Fermentable carbohydrates had no effect on colonic DNA damage. Dietary combinations of red meat with potato fiber or potato-resistant starch have distinctive effects in the large bowel. Future studies are essential to examine the efficacy of different types of nondigestible carbohydrates in maintaining colonic health during long-term consumption of high-protein diets. PRACTICAL APPLICATION: Improved understanding of interactions between the food consumed and gut microbiota provides knowledge needed to make healthier food choices for large bowel health. The impact of red meat on large bowel health may be ameliorated by consuming with fermentable dietary fiber, a colonic energy source that produces less harmful by-products than the microbial breakdown of colonic protein for energy. Developing functional red meat products with fermentable dietary fiber could be one way to promote a healthy and balanced macronutrient diet.


Assuntos
Biomarcadores/análise , Colo/metabolismo , Fibras na Dieta/análise , Solanum tuberosum/química , Amido/química , Ração Animal , Animais , Bacteroides fragilis/crescimento & desenvolvimento , Bacteroides fragilis/isolamento & purificação , Bifidobacterium/crescimento & desenvolvimento , Bifidobacterium/isolamento & purificação , Biomarcadores/metabolismo , Caseínas/administração & dosagem , Bovinos , Ceco/metabolismo , Celulose/administração & dosagem , Clostridium/crescimento & desenvolvimento , Clostridium/isolamento & purificação , Colo/microbiologia , Ensaio Cometa , Cresóis/análise , Dano ao DNA , DNA Bacteriano/genética , Dieta , Carboidratos da Dieta/análise , Enterococcus faecalis/crescimento & desenvolvimento , Enterococcus faecalis/isolamento & purificação , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/isolamento & purificação , Ácidos Graxos Voláteis/análise , Fezes/química , Fermentação , Intestino Grosso/metabolismo , Limosilactobacillus reuteri/crescimento & desenvolvimento , Limosilactobacillus reuteri/isolamento & purificação , Masculino , Carne , Metagenoma , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real
14.
Plant Foods Hum Nutr ; 67(3): 200-7, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22576129

RESUMO

We examined the effects of whole kiwifruit on gut microbiota using an in vitro batch model of gastric-ileal digestion and colonic fermentation. Faecal fermentations of gold and green kiwifruit, inulin and water (control) digests were performed for up to 48 h. As compared to the control, gold and green kiwifruit increased Bifidobacterium spp. by 0.9 and 0.8 log(10) cfu/ml, respectively (P < 0.001), and the Bacteroides-Prevotella-Porphyromonas group by 0.4 and 0.5 log(10) cfu/ml, respectively. Inulin only had a bifidogenic effect (+0.4 log(10) cfu/ml). This was accompanied with increases in microbial glycosidases, especially those with substrate specificities relating to the breakdown of kiwifruit oligosaccharides, and with increased generation of short chain fatty acids. The microbial metabolic activity was sustained for up to 48 h, which we attribute to the complexity of the carbohydrate substrate provided by whole kiwifruit. Kiwifruit fermenta supernatant was also separately shown to affect the in vitro proliferation of Bifidobacterium longum, and its adhesion to Caco-2 intestinal epithelial cells. Collectively, these data suggest that whole kiwifruit may modulate human gut microbial composition and metabolism to produce metabolites conducive to increased bifidobacteria-host association.


Assuntos
Actinidia/química , Bactérias/efeitos dos fármacos , Bifidobacterium/efeitos dos fármacos , Colo/efeitos dos fármacos , Frutas/química , Oligossacarídeos/farmacologia , Prebióticos , Adulto , Bactérias/crescimento & desenvolvimento , Bactérias/metabolismo , Aderência Bacteriana/efeitos dos fármacos , Bifidobacterium/crescimento & desenvolvimento , Bifidobacterium/metabolismo , Células CACO-2 , Colo/metabolismo , Colo/microbiologia , Ácidos Graxos Voláteis/metabolismo , Fezes/microbiologia , Feminino , Fermentação , Mucosa Gástrica/metabolismo , Glicosídeo Hidrolases/metabolismo , Humanos , Íleo/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Inulina/farmacologia , Masculino , Metagenoma/efeitos dos fármacos , Pessoa de Meia-Idade , Oligossacarídeos/metabolismo , Especificidade por Substrato
15.
Proteomics ; 12(4-5): 651-65, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22246638

RESUMO

Biofilms are surface-attached, matrix-encased, structured microbial communities which display phenotypic features that are dramatically different from those of their free-floating, or planktonic, counterparts. Biofilms seem to be the preferred mode of growth of microorganisms in nature, and at least 65% of all human infections are associated with biofilms. The most notable and clinically relevant property of biofilms is their greater resistance to antimicrobials compared with their planktonic counterparts. Although both bacterial and fungal biofilms display this phenotypic feature, the exact mechanisms underlying their increased drug resistance are yet to be determined. Advances in proteomics techniques during the past decade have facilitated in-depth analysis of the possible mechanisms underpinning increased drug resistance in biofilms. These studies have demonstrated the ability of proteomics techniques to unravel new targets for combating microbial biofilms. In this review, we discuss the putative drug resistance mechanisms of microbial biofilms that have been uncovered by proteomics and critically evaluate the possible contribution of the new knowledge to future development in the field. We also summarize strategic uses of novel proteomics technologies in studies related to drug resistance mechanisms of microbial biofilms.


Assuntos
Bactérias/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , Farmacorresistência Fúngica , Farmacorresistência Bacteriana Múltipla , Fungos/efeitos dos fármacos , Proteômica/métodos , Infecções Bacterianas/tratamento farmacológico , Infecções Bacterianas/metabolismo , Infecções Bacterianas/microbiologia , Fenômenos Fisiológicos Bacterianos , Biofilmes/crescimento & desenvolvimento , Fungos/fisiologia , Humanos , Estresse Oxidativo , Percepção de Quorum
16.
J Clin Periodontol ; 38(8): 694-701, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21752043

RESUMO

AIM: Porphyromonas gingivalis lipopolysaccharide (LPS) displays a significant amount of structural heterogeneity, containing both tetra- (LPS(1435/1449) ) and penta-acylated (LPS(1690) ) lipid A structures. This study investigated the effects of the two isoforms of P. gingivalis LPS on the expression of IL-6, IL-8 and TNF-α in human gingival fibroblasts (HGFs). MATERIALS AND METHODS: HGFs were stimulated with P. gingivalis LPS(1435/1449) and LPS(1690) in both dose- (1 ng-10 µg/ml) and time-dependent (2-48 h) experiments. Total RNA and protein were extracted and used for analysis of the IL-6, IL-8 and TNF-α transcripts as well as IL-6 and IL-8 proteins, by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. RESULTS: P. gingivalis LPS(1690) significantly up-regulated the mRNA and protein expression of IL-6 and IL-8, whereas P. gingivalis LPS(1435/1449) did not induce significant host response. The expression levels of IL-6 and IL-8 up-regulated by P. gingivalis LPS(1690) continuously increased with time course. In contrast, TNF-α transcript expression was up-regulated promptly by P. gingivalis LPS(1690) after 2 h of stimulation and gradually declined afterwards. CONCLUSIONS: This study suggests that P. gingivalis LPS heterogeneity may differentially modulate the pro-inflammatory cytokine expression in HGFs, which may contribute to periodontal pathogenesis.


Assuntos
Fibroblastos/imunologia , Gengiva/imunologia , Fatores Imunológicos/imunologia , Interleucina-6/imunologia , Interleucina-8/imunologia , Lipídeo A/imunologia , Porphyromonas gingivalis/imunologia , Linhagem Celular , Relação Dose-Resposta a Droga , Escherichia coli/imunologia , Gengiva/citologia , Humanos , Imunidade Inata/imunologia , Mediadores da Inflamação/imunologia , Lipídeo A/química , Proteínas/análise , RNA Mensageiro/análise , Fatores de Tempo , Fator de Necrose Tumoral alfa/imunologia , Regulação para Cima
17.
Int J Paediatr Dent ; 21(4): 299-305, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21401748

RESUMO

OBJECTIVES: Prunus mume is a common fruit in Asia, which has been used in traditional Chinese medicine. In this study, we focused on the antimicrobial properties of Prunus mume extract against oral pathogens related to dental caries and periodontal diseases. STUDY DESIGN: A total of 15 oral pathogens including Streptococcus mutans, S. sobrinus, S. mitis, S. sanguinis, Lactobacillus acidophilus, P. gingivalis, Aggregatibacter actinomycetemcomitans, and Candida species were included in the study. Initially, agar diffusion assay was performed to screen the antimicrobial activities of Prunus mume extract. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were then determined for sensitive species. Effect of Prunus mume extract on human oral keratinocytes (HOK) viability was also tested. RESULT: In the agar diffusion assay, drug suspension of 2 g/mL was able to inhibit all the bacterial species tested, but not the fungal species. MIC and MBC range of Prunus mume extract against the oral bacteria was 0.15625-0.0003 g/mL and P. gingivalis being the most susceptible species. Prune extract did not cause any detrimental effect on HOK. CONCLUSION: Prunus mume extract may be a potential candidate for developing an oral antimicrobial agent to control or prevent dental diseases associated with oral pathogenic bacteria.


Assuntos
Anti-Infecciosos/farmacologia , Cárie Dentária/microbiologia , Doenças Periodontais/microbiologia , Extratos Vegetais/farmacologia , Prunus , Aggregatibacter actinomycetemcomitans/efeitos dos fármacos , Candida/classificação , Candida/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Candida glabrata/efeitos dos fármacos , Candida tropicalis/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Ácido Cítrico/análise , Ácido Cítrico/farmacologia , Humanos , Queratinócitos/efeitos dos fármacos , Lactobacillus acidophilus/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Ácido Oxálico/análise , Ácido Oxálico/farmacologia , Extratos Vegetais/análise , Porphyromonas gingivalis/efeitos dos fármacos , Prunus/química , Saccharomyces cerevisiae/efeitos dos fármacos , Streptococcus/classificação , Streptococcus/efeitos dos fármacos , Streptococcus mitis/efeitos dos fármacos , Streptococcus mutans/efeitos dos fármacos , Streptococcus sobrinus/efeitos dos fármacos , Tartaratos/análise , Tartaratos/farmacologia
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