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1.
J. oral res. (Impresa) ; 7(6): 232-235, ago. 1, 2018. tab, graf
Article de Anglais | LILACS | ID: biblio-1120975

RÉSUMÉ

Objective: to compare the gene expression levels of collagen type I alpha 2 (COL1A2) in children with and without dental fluorosis. methods: cross-sectional study involving 92 children between 5 and 12 years of age. socio-demographic characteristics, the presence of dental fluorosis by means of the Thylstrup-Fejerskov index, and gene expression analysis of COL1A2 in peripheral blood samples by reverse transcription polymerase chain reaction (RT-PCR) assays, were described. for the descriptive analysis, measures of central tendency, dispersion and proportions were used. differences between the groups (p<0.05) were established by the student t-test. results: mean age was 8.6 (SD=1.9) years, 51.1 percent were female; 54 children were diagnosed with fluorosis and 38 without fluorosis; prevalence of dental fluorosis was 58.7 percent (95 percent CI: 48.4 percent -68.9 percent). gene expression of COL1A2 was statistically significantly lower (p<0.05) in the participants with dental fluorosis. conclusion: there are differences in the expression levels of the COL1A2 gene among the population under study. therefore, COL1A2 may be potentially involved in the development of dental fluorosis.


Sujet(s)
Humains , Mâle , Femelle , Enfant d'âge préscolaire , Enfant , Collagène de type I/physiologie , Fluorose dentaire/étiologie , Expression des gènes , Régulation de l'expression des gènes/physiologie , Études transversales , Colombie/épidémiologie
2.
Biol. Res ; 50: 43, 2017. tab, graf
Article de Anglais | LILACS | ID: biblio-950890

RÉSUMÉ

BACKGROUND: Understanding the molecular basis underlying the formation of bone-forming osteocytes and lipid-storing adipocytes will help provide insights into the cause of disorders originating in stem/progenitor cells and develop therapeutic treatments for bone- or adipose-related diseases. In this study, the role of RGS2 and RGS4, two members of the regulators of G protein signaling (RGS) family, was investigated during adipogenenic and osteogenenic differentiation of human mesenchymal stem cells (hMSCs). RESULTS: Expression of RGS2 and RGS4 were found to be inversely regulated during adipogenesis induced by dexamethasone (DEX) and 3-isobutyl-methylxanthine, regardless if insulin was present, with RGS2 up-regulated and RGS4 down-regulated in response to adipogenic induction. RGS2 expression was also up-regulated during osteogenesis at a level similar to that induced by treatment of DEX alone, a shared component of adipogenic and osteogenic differentiation inducing media, but significantly lower than the level induced by adipogenic inducing media. RGS4 expression was down-regulated during the first 48 h of osteogenesis but up-regulated afterwards, in both cases at levels similar to that induced by DEX alone. Expression knock-down using small interfering RNA against RGS2 resulted in decreased differentiation efficiency during both adipogenesis and osteogenesis. On the other hand, expression knock-down of RGS4 also resulted in decreased adipogenic differentiation but increased osteogenic differentiation. CONCLUSIONS: RGS2 and RGS4 are differentially regulated during adipogenic and osteogenic differentiation of hMSCs. In addition, both RGS2 and RGS4 play positive roles during adipogenesis but opposing roles during osteogenesis, with RGS2 as a positive regulator and RGS4 as a negative regulator. These results imply that members of RGS proteins may play multifaceted roles during human adipogenesis and osteogenesis to balance or counterbalance each other's function during those processes.


Sujet(s)
Humains , Ostéocytes/cytologie , Ostéogenèse/physiologie , Régulation de l'expression des gènes/physiologie , Protéines RGS/métabolisme , Adipogenèse/physiologie , Cellules souches mésenchymateuses/cytologie , Ostéogenèse/génétique , Facteurs temps , Régulation de l'expression des gènes/génétique , Protéines RGS/génétique , Adipogenèse/génétique
3.
Mem. Inst. Oswaldo Cruz ; 111(11): 686-691, Nov. 2016. graf
Article de Anglais | LILACS | ID: biblio-829250

RÉSUMÉ

Trypanosomatids are parasites that cause disease in humans, animals, and plants. Most are non-pathogenic and some harbor a symbiotic bacterium. Endosymbiosis is part of the evolutionary process of vital cell functions such as respiration and photosynthesis. Angomonas deanei is an example of a symbiont-containing trypanosomatid. In this paper, we sought to investigate how symbionts influence host cells by characterising and comparing the transcriptomes of the symbiont-containing A. deanei (wild type) and the symbiont-free aposymbiotic strains. The comparison revealed that the presence of the symbiont modulates several differentially expressed genes. Empirical analysis of differential gene expression showed that 216 of the 7625 modulated genes were significantly changed. Finally, gene set enrichment analysis revealed that the largest categories of genes that downregulated in the absence of the symbiont were those involved in oxidation-reduction process, ATP hydrolysis coupled proton transport and glycolysis. In contrast, among the upregulated gene categories were those involved in proteolysis, microtubule-based movement, and cellular metabolic process. Our results provide valuable information for dissecting the mechanism of endosymbiosis in A. deanei.


Sujet(s)
Humains , Animaux , Régulation de l'expression des gènes/physiologie , Gene Ontology , ARN des protozoaires/génétique , Symbiose/génétique , Transcriptome/génétique , Trypanosomatina/génétique , Bactéries/croissance et développement , Analyse de profil d'expression de gènes , Gènes de protozoaire , Génome de protozoaire , Génomique , ARN des protozoaires/isolement et purification , Trypanosomatina/métabolisme
4.
Salud colect ; 11(1): 67-86, ene.-mar. 2015. ilus
Article de Espagnol | LILACS | ID: lil-746685

RÉSUMÉ

Este artículo explica las dificultades que tienen las farmacéuticas innovadoras para retribuir a sus accionistas con dividendos atractivos. El problema es el resultado de la caducidad de las patentes de los medicamentos estrella (blockbusters) y las dificultades que tienen en desarrollar nuevos medicamentos estrella. Una solución que las empresas han encontrado es acelerar la ejecución de los ensayos clínicos para obtener, en el menor tiempo posible, el permiso de comercialización y así incrementar el tiempo monopólico de ventas de los nuevos medicamentos. En este contexto, los autores describen la forma en que las farmacéuticas innovadoras acortan el tiempo de ejecución de los ensayos en América Latina y las consecuencias en la calidad de los datos que se obtienen, en la protección de los derechos humanos de los sujetos de experimentación, y en el cumplimiento de los principios éticos aprobados en las declaraciones universales.


This article explains the difficulties innovative pharmaceutical firms have in repaying shareholders with attractive dividends. The problem is the result of the expiration of the patents of blockbuster drugs and the difficulties that the firms have in bringing new blockbuster drugs to the market. One of the solutions companies have found has been to accelerate the implementation of clinical trials in order to expedite the commercialization of new drugs. Doing so increases the period in which they can sell drugs at monopoly prices. We therefore discuss how innovative pharmaceutical firms shorten the implementation time of clinical trials in Latin America and the consequences such actions have on the quality of the collected data, the protection of human rights of the subjects of experimentation, and compliance with the ethical principles approved in international declarations.


Sujet(s)
Animaux , Souris , Algorithmes , Mouvement cellulaire/physiologie , Techniques d'aide à la décision , Protéines de la matrice extracellulaire/métabolisme , Régulation de l'expression des gènes/physiologie , Modèles biologiques , Transduction du signal/physiologie , Simulation numérique , Modèles logistiques
6.
Korean J. Ophthalmol ; Korean J. Ophthalmol;: 424-432, 2015.
Article de Anglais | WPRIM | ID: wpr-219865

RÉSUMÉ

PURPOSE: To evaluate the effects of bevacizumab on expression of B-cell leukemia/lymphoma (Bcl)-2 and apoptosis in retinal pigment epithelial (RPE) cells under oxidative stress conditions. METHODS: RPE cells were treated with H2O2 (0, 100, 200, 300, and 400 microM) and bevacizumab at or above the doses normally used in clinical practice (0, 0.33, 0.67, 1.33, and 2.67 mg/mL). Cell apoptosis was measured using flow cytometry with annexin V-fluorescein isothiocyanate. The expression of Bcl-2 mRNA was determined using reverse transcription polymerase chain reaction. RESULTS: Under low oxidative stress conditions (H2O2 100 microM), cell apoptosis was not significantly different at any concentration of bevacizumab, but Bcl-2 mRNA expression decreased with increasing concentration of bevacizumab (0.33, 0.67, 1.33, and 2.67 mg/mL). Under moderate oxidative stress conditions (H2O2 200 microM), Bcl-2 mRNA expression decreased with increasing concentration of bevacizumab (0.33, 0.67, 1.33, and 2.67 mg/mL), but cell apoptosis increased only at 2.67 mg/mL of bevacizumab. Under high oxidative stress (300 microM) conditions, cell apoptosis increased at high concentrations of bevacizumab (1.33 and 2.67 mg/mL), but it did not correlate with Bcl-2 expression. CONCLUSIONS: Withdrawal of vascular endothelial growth factor can lead to RPE cell apoptosis and influences the expression of anti-apoptotic genes such as Bcl-2 under oxidative stress conditions. Since oxidative stress levels of each patient are unknown, repeated injections of intravitreal bevacizumab, as in eyes with age-related macular degeneration, might influence RPE cell survival.


Sujet(s)
Humains , Inhibiteurs de l'angiogenèse/pharmacologie , Apoptose/effets des médicaments et des substances chimiques , Bévacizumab/pharmacologie , Lignée cellulaire , Test ELISA , Cytométrie en flux , Régulation de l'expression des gènes/physiologie , Peroxyde d'hydrogène/toxicité , Stress oxydatif/effets des médicaments et des substances chimiques , Protéines proto-oncogènes c-bcl-2/génétique , ARN messager/génétique , Réaction de polymérisation en chaine en temps réel , Épithélium pigmentaire de la rétine/effets des médicaments et des substances chimiques , Facteur de croissance endothéliale vasculaire de type A/antagonistes et inhibiteurs
7.
J. vet. sci ; J. vet. sci;: 37-46, 2015.
Article de Anglais | WPRIM | ID: wpr-206914

RÉSUMÉ

Toll-like receptor 5 (TLR5) is responsible for the recognition of bacterial flagellin in vertebrates. In the present study, the first TLR5 gene in duck was cloned. The open reading frame (ORF) of duck TLR5 (dTLR5) cDNA is 2580 bp and encodes a polypeptide of 859 amino acids. We also cloned partial sequences of myeloid differentiation factor 88, 2'-5'-oligoadenylate synthetase (OAS), and myxovirus resistance (Mx) genes from duck. dTLR5 mRNA was highly expressed in the bursa of Fabricius, spleen, trachea, lung, jejunum, rectum, and skin; moderately expressed in the muscular and glandular tissues, duodenum, ileum, caecum, and pancreas; and minimally expressed in the heart, liver, kidney, and muscle. DF-1 or HeLa cells transfected with DNA constructs encoding dTLR5 can activate NF-kappaB leading to the activation of interleukin-6 (IL-6) promoter. When we challenged ducks with a Herts33 Newcastle disease virus (NDV), mRNA transcription of the antiviral molecules Mx, Double stranded RNA activated protein kinase (PKR), and OAS was up-regulated in the liver, lung, and spleen 1 and 2 days post-inoculation.


Sujet(s)
Animaux , Humains , 2',5'-Oligoadenylate synthetase/génétique , Lignée cellulaire , Clonage moléculaire , Canards , Régulation de l'expression des gènes/physiologie , Immunité innée , Facteur de différenciation myéloïde-88/génétique , Protéines de résistance aux myxovirus/génétique , Maladie de Newcastle/métabolisme , Virus de la maladie de Newcastle/classification , ARN messager/génétique , Spécificité d'espèce , Récepteur de type Toll-5/génétique
8.
Rev. Esc. Enferm. USP ; Rev. Esc. Enferm. USP;48(spe): 53-58, 08/2014.
Article de Anglais | LILACS, BDENF | ID: lil-731286

RÉSUMÉ

Objective To understand the experiences and expectations of nurses in the treatment of women with chronic venous ulcers. Method Phenomenological research was based on Alfred Schütz, whose statements were obtained in January, 2012, through semi-structured interviews with seven nurses. Results The nurse reveals the difficulties presented by the woman in performing self-care, the perceived limitations in the treatment anchored in motivation, and the values and beliefs of women. It showed professional frustration because venous leg ulcer recurrence, lack of inputs, interdisciplinary work and training of nursing staff. There was an expected adherence to the treatment of women, and it emphasized the need for ongoing care, supported self-care and standard practices in treatment. Conclusion That treatment of chronic venous leg ulcers constitutes a challenge that requires collective investment, involving women, professionals, managers and health institutions. .


Objetivo Comprender las experiencias y expectativas de enfermeras en el tratamiento de mujeres con úlcera venosa crónica. Método Investigación fenomenológica fundamentada en Alfred Schutz, que buscó Se realizó entrevista semiestructurada con siete enfermeras, en enero del 2012. Resultados La enfermera revela dificultades presentadas por la mujer para realizar el autocuidado, percibe limitaciones en el tratamiento relacionadas con la desmotivación, los valores y las creencias de las mujeres. Refiere frustración profesional debido a la recidiva de la lesión, a la falta de insumos, al deficiente trabajo interdisciplinar y a la limitada capacitación del equipo de enfermeras. Espera la adhesión de la mujer al tratamiento y resalta la necesidad del cuidado continuo, del autocuidado apoyado y de estandarizar conductas de tratamiento. Conclusión El tratamiento de la úlcera venosa crónica es un desafío que requiere contribución colectiva, involucrando a las mujeres, a los profesionales, a los gestores y a las instituciones de salud. .


Objetivo Compreender as experiências e expectativas de enfermeiras no tratamento de mulheres com úlcera venosa crônica na Atenção Primária à Saúde. Método Pesquisa fundamentada na fenomenologia social de Alfred Schütz, com depoimentos obtidos em janeiro de 2012, por meio de entrevista semiestruturada com sete enfermeiras. Resultados As enfermeiras revelam dificuldades apresentadas pelas mulheres com úlcera venosa crônica para realizar o autocuidado, percebem limitações na terapêutica ancoradas na desmotivação e nos valores e crenças das mulheres. Referem frustração profissional em razão da recidiva da lesão, falta de insumos e tecnologia, de trabalho interdisciplinar e da capacitação da equipe de enfermagem. Esperam a adesão das mulheres ao tratamento e ressaltam a necessidade do cuidado contínuo, do autocuidado apoiado e da padronização de condutas no tratamento. Conclusão O tratamento da úlcera venosa crônica constitui-se em um desafio que requer investimento coletivo, envolvendo a mulher, os profissionais, os gestores e as instituições de saúde. .


Sujet(s)
Animaux , Protéines de Caenorhabditis elegans/isolement et purification , Caenorhabditis elegans/métabolisme , Membrane cellulaire/métabolisme , Canaux ioniques/isolement et purification , Canaux ioniques/métabolisme , Protéines de tissu nerveux/isolement et purification , Protéines de tissu nerveux/métabolisme , Système nerveux/métabolisme , Neurones afférents/métabolisme , Sensation/génétique , Séquence d'acides aminés/génétique , Séquence nucléotidique/génétique , Comportement animal/effets des médicaments et des substances chimiques , Comportement animal/physiologie , Protéines de Caenorhabditis elegans/génétique , Protéines de Caenorhabditis elegans/métabolisme , Caenorhabditis elegans/cytologie , Capsaïcine/pharmacologie , Compartimentation cellulaire/génétique , Membrane cellulaire/effets des médicaments et des substances chimiques , Membrane cellulaire/ultrastructure , Régulation de l'expression des gènes/physiologie , Canaux ioniques/génétique , Canaux ioniques/ultrastructure , Données de séquences moléculaires , Mutation/génétique , Protéines de tissu nerveux/génétique , Protéines de tissu nerveux/ultrastructure , Système nerveux/cytologie , Système nerveux/effets des médicaments et des substances chimiques , Neurones afférents/cytologie , Neurones afférents/effets des médicaments et des substances chimiques , Douleur/génétique , Douleur/métabolisme , Douleur/physiopathologie , Phylogenèse , Récepteurs des médicaments/effets des médicaments et des substances chimiques , Récepteurs des médicaments/métabolisme , Récepteurs des médicaments/ultrastructure , Sensation/effets des médicaments et des substances chimiques , Transduction du signal/génétique , Canaux cationiques TRPV , Canaux cationiques TRP
9.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;47(4): 279-286, 8/4/2014. tab, graf
Article de Anglais | LILACS | ID: lil-705770

RÉSUMÉ

SRY-related high-mobility-group box 9 (Sox9) gene is a cartilage-specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. The aim of this study was to investigate the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUC-MSCs). After they were isolated from human umbilical cord blood within 24 h after delivery of neonates, hUC-MSCs were untreated or transfected with a human Sox9-expressing plasmid or an empty vector. The cells were assessed for morphology and chondrogenic differentiation. The isolated cells with a fibroblast-like morphology in monolayer culture were positive for the MSC markers CD44, CD105, CD73, and CD90, but negative for the differentiation markers CD34, CD45, CD19, CD14, or major histocompatibility complex class II. Sox9 overexpression induced accumulation of sulfated proteoglycans, without altering the cellular morphology. Immunocytochemistry demonstrated that genetic delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in hUC-MSCs compared with empty vector-transfected counterparts. Reverse transcription-polymerase chain reaction analysis further confirmed the elevation of aggrecan and type II collagen at the mRNA level in Sox9-transfected cells. Taken together, short-term Sox9 overexpression facilitates chondrogenesis of hUC-MSCs and may thus have potential implications in cartilage tissue engineering.


Sujet(s)
Humains , Différenciation cellulaire/génétique , Chondrogenèse/génétique , Sang foetal/cytologie , Cellules souches mésenchymateuses/cytologie , Facteur de transcription SOX-9/génétique , Agrécanes/biosynthèse , Technique de Western , Cartilage/métabolisme , Prolifération cellulaire/génétique , Chondrocytes/métabolisme , Collagène de type II/biosynthèse , Cytométrie en flux , Protéines à fluorescence verte , Régulation de l'expression des gènes/physiologie , Cellules endothéliales de la veine ombilicale humaine/cytologie , Immunohistochimie , Immunophénotypage , Culture de cellules primaires , RT-PCR , Ingénierie tissulaire , Transfection
10.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;47(2): 101-109, 2/2014. tab, graf
Article de Anglais | LILACS | ID: lil-699773

RÉSUMÉ

In the current literature, there is evidence that psychological factors can affect the incidence and progression of some cancers. Interleukin 6 (IL-6) is known to be elevated in individuals experiencing chronic stress and is also involved in oncogenesis and cancer progression. However, the precise mechanism of IL-6 induction by the stress-related hormone norepinephrine (NE) is not clear, and, furthermore, there are no reports about the effect of NE on IL-6 expression in gastric epithelial cells. In this study, we examined the effect of NE on IL-6 expression in immortalized human gastric epithelial cells (GES-1 cells). Using real-time PCR and enzyme-linked immunoassay, we demonstrated that NE can induce IL-6 mRNA and protein expression in GES-1 cells. The induction is through the β-adrenergic receptor-cAMP-protein kinase A pathway and mainly at the transcriptional level. Progressive 5′-deletions and site-directed mutagenesis of the parental construct show that, although activating-protein-1 (AP-1), cAMP-responsive element binding protein (CREB), CCAAT-enhancer binding protein-β (C/EBP-β), and nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB) binding sites are all required in the basal transcription of IL-6, only AP-1 and CREB binding sites in the IL-6 promoter are required in NE-induced IL-6 expression. The results suggest that chronic stress may increase IL-6 secretion of human gastric epithelial cells, at least in part, by the stress-associated hormone norepinephrine, and provides basic data on stress and gastric cancer progression.


Sujet(s)
Humains , Cellules épithéliales/effets des médicaments et des substances chimiques , /métabolisme , Norépinéphrine/pharmacologie , Transduction du signal/physiologie , Lignée cellulaire , Test ELISA , Cellules épithéliales/métabolisme , Muqueuse gastrique/effets des médicaments et des substances chimiques , Muqueuse gastrique/métabolisme , Régulation de l'expression des gènes/physiologie , /génétique , Facteur de transcription NF-kappa B/métabolisme , Norépinéphrine/métabolisme , Réaction de polymérisation en chaine en temps réel , ARN messager/métabolisme , Récepteurs bêta-adrénergiques/métabolisme , Facteurs de transcription/physiologie , Régulation positive
11.
Korean J. Ophthalmol ; Korean J. Ophthalmol;: 343-350, 2014.
Article de Anglais | WPRIM | ID: wpr-156974

RÉSUMÉ

PURPOSE: To investigate the relationship between plasma TDRD7 mRNA and lens transparency, and to evaluate plasma TDRD7 mRNA as a potential marker for cataracts and its sub-type by quantitatively analyzing human peripheral blood. METHODS: Plasma RNA was extracted from 40 patients with cataracts, and 30 normal controls of matched age and gender. Blood cholesterol and fasting glucose were measured, and the RNA extracted from the sample was synthesized into cDNA. After polymerase chain reaction, the results were compared after quantifying the TDRD7 mRNA using ABL1 mRNA for normalization. We analyzed the relative gene expression data via the DeltaDeltaCt method. RESULTS: The normalized 2(-DeltaDeltaCt) of plasma TDRD7 mRNA based on ABL1 mRNA was 1.52 ± 0.63 in the case of the control group and 1.05 ± 0.34 in the case of the cataract patients, and the TDRD7 expression level of the cataract patients was lower than that of the control group (p = 0.048). The comparison of the genetic values of different types of cataracts demonstrated that the TDRD7 expression level of the cortical type and mixed type were lower than those of the nuclear type and posterior subcapsular opacity type (p = 0.017). CONCLUSIONS: Human cataracts and the TDRD7 gene loss-of-function mutations are strongly causally related, as the expression level of plasma TDRD7 mRNA in patients with cataracts was statistically significantly lower than in the normal control group.


Sujet(s)
Adolescent , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Enfant , Femelle , Humains , Mâle , Adulte d'âge moyen , Cataracte/sang , Régulation de l'expression des gènes/physiologie , Protéines proto-oncogènes c-abl/génétique , ARN messager/sang , Réaction de polymérisation en chaine en temps réel , Ribonucléoprotéines/génétique
12.
Biol. Res ; 47: 1-7, 2014. ilus, graf, tab
Article de Anglais | LILACS | ID: biblio-950768

RÉSUMÉ

BACKGROUND: Acetylcholine (ACh) is known to be a key neurotransmitter in the central and peripheral nervous systems, which is also produced in a variety of non-neuronal tissues and cell. The existence of ACh in maxilla in vivo and potential regulation role for osteogenesis need further study. RESULTS: Components of the cholinergic system (ACh, esterase, choline acetyltransferase, high-affinity choline uptake, n- and mAChRs) were determined in maxilla of rat in vivo, by means of Real-Time PCR and immunohistochemistry. Results showed RNA for CarAT, carnitine/acylcarnitine translocase member 20 (Slc25a20), VAChT, OCTN2, OCT1, OCT3, organic cation transporter member 4 (Slc22a4), AChE, BChE, nAChR subunits α1, α2, α3, α5, α7, α10, ß1, ß2, ß4, γ and mAChR subunits M1, M2, M3, M4, M5 were detected in rat's maxilla. RNA of VAChT, AChE, nAChR subunits α2, ß1, ß4 and mAChR subunits M4 had abundant expression (2(-ΔCt) > 0.03). Immunohistochemical staining was conducted for ACh, VAChT, nAChRα7 and AChE. ACh was expressed in mesenchymal cells, chondroblast, bone and cartilage matrix and bone marrow cells, The VAChT expression was very extensively while ACh receptor α7 was strongly expressed in newly formed bone matrix of endochondral and bone marrow ossification, AchE was found only in mesenchymal stem cells, cartilage and bone marrow cells. CONCLUSIONS: ACh might exert its effect on the endochondral and bone marrow ossification, and bone matrix mineralization in maxilla.


Sujet(s)
Animaux , Mâle , Rats , Moelle osseuse/physiologie , Acétylcholine/métabolisme , Cartilage/physiologie , Agents cholinergiques/métabolisme , Maxillaire/métabolisme , Ostéogenèse/physiologie , Trame osseuse/métabolisme , Calcification physiologique/physiologie , Cellules de la moelle osseuse/métabolisme , Immunohistochimie , Carnitine acyltransferases/génétique , Carnitine acyltransferases/métabolisme , Régulation de l'expression des gènes/physiologie , Récepteurs nicotiniques/génétique , Rat Sprague-Dawley , Transporteurs de cations organiques/génétique , Transporteurs de cations organiques/métabolisme , Transporteurs vésiculaires de l'acétylcholine/génétique , Transporteurs vésiculaires de l'acétylcholine/métabolisme , Cellules souches mésenchymateuses/métabolisme , Réaction de polymérisation en chaine en temps réel , Maxillaire/cytologie
13.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;46(3): 287-292, 15/mar. 2013. tab, graf
Article de Anglais | LILACS | ID: lil-670902

RÉSUMÉ

This study investigated the consequences of intrauterine protein restriction on the gastrointestinal tract and particularly on the gene expression and activity of intestinal disaccharidases in the adult offspring. Wistar rat dams were fed isocaloric diets containing 6% protein (restricted, n = 8) or 17% protein (control, n = 8) throughout gestation. Male offspring (n = 5-8 in each group) were evaluated at 3 or 16 weeks of age. Maternal protein restriction during pregnancy produced offspring with growth restriction from birth (5.7 ± 0.1 vs 6.3 ± 0.1 g; mean ± SE) to weaning (42.4 ± 1.3 vs 49.1 ± 1.6 g), although at 16 weeks of age their body weight was similar to control (421.7 ± 8.9 and 428.5 ± 8.5 g). Maternal protein restriction also increased lactase activity in the proximal (0.23 ± 0.02 vs 0.15 ± 0.02), medial (0.30 ± 0.06 vs 0.14 ± 0.01) and distal (0.43 ± 0.07 vs 0.07 ± 0.02 U·g-1·min-1) small intestine, and mRNA lactase abundance in the proximal intestine (7.96 ± 1.11 vs 2.38 ± 0.47 relative units) of 3-week-old offspring rats. In addition, maternal protein restriction increased sucrase activity (1.20 ± 0.02 vs 0.91 ± 0.02 U·g-1·min-1) and sucrase mRNA abundance (4.48 ± 0.51 vs 1.95 ± 0.17 relative units) in the duodenum of 16-week-old rats. In conclusion, the present study shows for the first time that intrauterine protein restriction affects gene expression of intestinal enzymes in offspring.


Sujet(s)
Animaux , Femelle , Grossesse , Régime pauvre en protéines , Disaccharidases/métabolisme , Régulation de l'expression des gènes/physiologie , Intestin grêle/enzymologie , Adaptation physiologique , Animaux nouveau-nés , Disaccharidases/analyse , Rat Wistar , Réaction de polymérisation en chaine en temps réel
14.
J. vet. sci ; J. vet. sci;: 69-76, 2013.
Article de Anglais | WPRIM | ID: wpr-219416

RÉSUMÉ

The use of mesenchymal stem cells (MSCs) has emerged as a potential new treatment for myocardial infarction. However, the poor viability of MSCs after transplantation critically limits the efficacy of this new strategy. The expression of microRNA-210 (miR-210) is induced by hypoxia and is important for cell survival under hypoxic conditions. Hypoxia increases the levels of hypoxia inducible factor-1 (HIF-1) protein and miR-210 in human MSCs (hMSCs). miR-210 positively regulates HIF-1alpha activity. Furthermore, miR-210 expression is also induced by hypoxia through the regulation of HIF-1alpha. To investigate the effect of miR-210 on hMSC survival under hypoxic conditions, survival rates along with signaling related to cell survival were evaluated in hMSCs over-expressing miR-210 or ones that lacked HIF-1alpha expression. Elevated miR-210 expression increased survival rates along with Akt and ERK activity in hMSCs with hypoxia. These data demonstrated that a positive feedback loop involving miR-210 and HIF-1alpha was important for MSC survival under hypoxic conditions.


Sujet(s)
Humains , Survie cellulaire , Cobalt , Régulation de l'expression des gènes/physiologie , Sous-unité alpha du facteur-1 induit par l'hypoxie/génétique , Cellules souches mésenchymateuses/effets des médicaments et des substances chimiques , microARN/métabolisme , Oxygène/pharmacologie , Consommation d'oxygène , Petit ARN interférent/métabolisme
15.
Article de Anglais | WPRIM | ID: wpr-79739

RÉSUMÉ

Vaccination is considered a promising alternative for controlling tick infestations. Haemaphysalis longicornis midgut proteins separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membrane were screened for protective value against bites. The western blot demonstrated the immunogenicity of 92 kDa protein (P92). The analysis of the P92 amino acid sequence by LC-MS/MS indicated that it was a H. longicornis paramyosin (Hl-Pmy). The full lenghth cDNA of Hl-Pmy was obtained by rapid amplification of cDNA ends (RACE) which consisted of 2,783 bp with a 161 bp 3' untranslated region. Sequence alignment of tick paramyosin (Pmy) showed that Hl-Pmy shared a high level of conservation among ticks. Comparison with the protective epitope sequence of other invertebrate Pmy, it was calculated that the protective epitope of Hl-Pmy was a peptide (LEEAEGSSETVVEMNKKRDTE) named LEE, which was close to the N-terminal of Hl-Pmy protein. The secondary structure analysis suggested that LEE had non-helical segments within an alpha-helical structure. These results provide the basis for developing a vaccine against biting H. longicornis ticks.


Sujet(s)
Animaux , Séquence d'acides aminés , Antigènes/génétique , Séquence nucléotidique , Technique de Western , Chromatographie en phase liquide , Clonage moléculaire , ADN complémentaire/génétique , Épitopes , Régulation de l'expression des gènes/physiologie , Ixodidae/génétique , Données de séquences moléculaires , Spectrométrie de masse en tandem
16.
Arq. bras. cardiol ; Arq. bras. cardiol;99(3): 780-788, set. 2012. ilus, tab
Article de Portugais | LILACS | ID: lil-649261

RÉSUMÉ

FUNDAMENTO: O programa de biogênese mitocondrial no coração parece apresentar remodelação adaptativa após estresse biomecânico e oxidativo. Os mecanismos adaptativos que protegem o metabolismo do miocárdio durante a hipóxia são coordenados, em parte, pelo óxido nítrico (NO). OBJETIVO: Observar a biogênese mitocondrial e expressão do óxido nítrico sintase (NOS) em corações de cardiopatia congênita com cianose; discutir a resposta mitocondrial à hipóxia crônica do miocárdio. MÉTODOS: Foram investigados 20 pacientes com defeitos cardíacos cianóticos (n = 10) ou acianóticos (n = 10). Foram estudadas amostras do miocárdio na via de saída ventricular direita, tomadas durante a operação. A análise morfométrica de mitocôndrias foi realizada por microscopia eletrônica de transmissão. A relação mtDNA/nDNA foi determinada com PCR em tempo real. Os níveis de transcrição da subunidade I da citocromo c oxidase (COXI), coativador-1α do receptor γ ativado por proliferador de peroxissoma (PGC-1α), o fator respiratório nuclear 1 (NRF1), e fator de transcrição mitocondrial A (Tfam) foram detectados por reação em cadeia da polimerase via transcriptase reversa (RT-PCR) ativado por fluorescência em tempo real. Os níveis proteicos de COXI e nNOS, iNOS e eNOS foram medidos por técnica de Western Blot. RESULTADOS: A densidade volumétrica mitocondrial (Vv) e a densidade numérica (Nv) foram significativamente elevadas em pacientes com cianose, em comparação com a cardiopatia congênita acianótica. MtDNA elevada e suprarregulação dos níveis de COXI, PGC-1 α, NRF1 e Tfam mRNA foram observadas em pacientes cianóticos. Os níveis de proteína de COXI e eNOS foram significativamente maiores no miocárdio de pacientes cianóticos que nos de acianóticos. Os níveis de transcrição do PGC-1α se correlacionam com os níveis de eNOS. CONCLUSÃO: A biogênese mitocondrial é ativada no miocárdio da via de saída ventricular na cardiopatia congênita com cianose, que ...


BACKGROUND: Mitochondrial biogenesis program in heart appears to exhibit adaptive remodeling following biomechanical and oxidative stress. The adaptive mechanisms that protect myocardium metabolism during hypoxia are coordinated in part by nitric oxide (NO). OBJECTIVE: To observe mitochondrial biogenesis and nitric oxide synthase (NOS) expression in hearts of congenital heart disease with cyanosis, discuss mitochondrial response to chronic hypoxia in myocardium. METHODS: 20 patients with cyanotic (n=10) or acyanotic cardiac defects (n=10) were investigated. Samples from the right ventricular outflow tract myocardium taken during operation were studied. Morphometric analysis of mitochondria was performed with transmission electron microscope. Relative mtDNA/nDNA ratio was determined with real-time PCR. Cytochrome c oxidase subunit I (COXI), peroxisome-proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF1), and mitochondrial transcription factor A (Tfam) transcript levels were detected by real-time fluorescent RT-PCR. COXI and nNOS, iNOS and eNOS protein levels were measured with western blot. RESULTS: Mitochondrial volume density (Vv) and numerical density (Nv) were significantly elevated in patients with cyanotic compared to acyanotic congenital heart disease. Elevated mtDNA and up-regulated COXI, PGC-1α, NRF1 and Tfam mRNA levels were observed in cyanotic patients. Protein levels of COXI and eNOS were significantly higher in the myocardium of cyanotic than of acyanotic patients. PGC-1α transcript levels correlated with the levels of eNOS. Conclusion: Mitochondrial biogenesis is activated in right ventricular outflow tract myocardium in congenital heart disease with cyanosis, which could be the adaptive response to chronic hypoxia and possibly involves eNOS up-regulation. (Arq Bras Cardiol. 2012; [online].ahead print, PP.0-0).


Sujet(s)
Adolescent , Enfant , Enfant d'âge préscolaire , Femelle , Humains , Mâle , Jeune adulte , Cyanose/enzymologie , Cyanose/physiopathologie , Cardiopathies congénitales/enzymologie , Renouvellement des mitochondries/physiologie , Myocarde/métabolisme , Nitric oxide synthase type III/métabolisme , Variations de nombre de copies de segment d'ADN , ADN mitochondrial/composition chimique , Régulation de l'expression des gènes/physiologie , Cardiopathies congénitales/physiopathologie , Taille de la mitochondrie , Nitric oxide synthase/métabolisme , RT-PCR , Facteurs de transcription/métabolisme
17.
J. vet. sci ; J. vet. sci;: 179-185, 2012.
Article de Anglais | WPRIM | ID: wpr-174786

RÉSUMÉ

The aim of this study was to demonstrate and assess C-reactive protein (CRP) changes in dogs with induced bacterial cystitis with or without antibiotics. We also evaluated availability of CRP levels to serve as an indicator for monitoring or diagnosing bacterial cystitis. Serial CRP concentrations in dogs with induced bacterial cystitis were higher than those of controls (p < 0.001). CRP concentrations peaked on day 7 and gradually decreased thereafter. In the treatment group, CRP concentrations decreased after medication compared to the untreated group (p = 0.032). CRP levels had a linear correlation with urine white blood cell counts among all groups (r = 0.837, p < 0.001, n = 140). Compared to the negative urine culture group, dogs with positive urine culture results had higher CRP concentrations (median 43.8 mg/L vs. 5.9 mg/L; p < 0.001). Area under the receiver operating characteristic curve was 0.955; when cut-off value was 12.2 mg/L, CRP measurements were found to have a sensitivity of 92.3% and specificity of 86.4%. This result indicates that rapid increases of CRP occurred after inducing bacterial cystitis and CRP may be a useful indicator for monitoring or diagnosing canine bacterial cystitis together with sediment urinalysis and urine bacterial culture.


Sujet(s)
Animaux , Chiens , Mâle , Association amoxicilline-clavulanate de potassium/usage thérapeutique , Antibactériens/usage thérapeutique , Protéine C-réactive/génétique , Cystite/métabolisme , Régulation de l'expression des gènes/physiologie , Inflammation/métabolisme , Infections à Proteus/traitement médicamenteux , Proteus mirabilis
18.
J. vet. sci ; J. vet. sci;: 163-170, 2012.
Article de Anglais | WPRIM | ID: wpr-174788

RÉSUMÉ

The purpose of this study was to measure the thickness of canine epidermis at various anatomical sites according to localization of cornified envelopes (involucrin and filaggrin), keratins (keratin 10, 5), and their mRNA expression. This was done in the skin of five breeds of dogs including seven poodles, six golden retrievers, six Shih Tzus, four pugs, and four Labrador retrievers. Epidermal thickness of the stratum corneum and nucleated epidermal layer was significantly different. The greatest thickness was observed in the digital web area and the thinnest epidermis was in the axilla. Epidermal thickness was also significantly different between the breeds (p < 0.05). Immunohistochemical staining scores revealed significant decreases of involucrin, filaggrin, and keratin 10 in the ventral and weight-bearing sites, and a relative increase of keratin 5 (p < 0.05). q-PCR analysis showed that their the levels of mRNA were positively correlated with expression of the corresponding proteins in skin samples (p < 0.05). The present study is the first to report the relationship between epidermal gene expression and histologic morphology of the skin in normal dogs. Further studies will be essential to fully understand the pathogenesis of skin barrier dysfunctions in canines.


Sujet(s)
Animaux , ADN complémentaire/génétique , Chiens/anatomie et histologie , Régulation de l'expression des gènes/physiologie , Protéines de filaments intermédiaires/génétique , Kératine-10/génétique , Kératine-5/génétique , Réaction de polymérisation en chaîne/méthodes , Précurseurs de protéines/génétique , ARN/génétique , Peau/anatomie et histologie
19.
J. vet. sci ; J. vet. sci;: 379-385, 2011.
Article de Anglais | WPRIM | ID: wpr-186143

RÉSUMÉ

Ectonucleotide pyrophosphatase/phosphodiestrase 2 (Enpp2) isolated from the supernatant of human melanoma cells is a lysophospholipase D that transforms lysophosphatidylcholine into lysophospatidic acid. Although multiple analyses have investigated the function of Enpp2 in the hypothalamus, its role in the uterus during the estrous cycle is not well understood. In the present study, rat uterine Enpp2 was analyzed by RT-PCR, Western blotting, and immunohistochemistry. Quantitative PCR analysis demonstrated that uterine Enpp2 mRNA was decreased during estrus compared to proestrus and diestrus. To determine whether uterine Enpp2 expression is affected by sex steroid hormones, immature rats were treated with 17beta-estradiol (E2), progesterone, or both on postnatal days 14 to 16. Interestingly, the expression of Enpp2 mRNA and protein were down-regulated by E2 in the uterus during estrus but not during proestrus or diestrus, suggesting that Enpp2 may play a role in uterine function during estrus. Enpp2 is primarily localized in the stromal cells of the endometrium during proestrus and estrus. During diestrus, Enpp2 was highly expressed in the epithelial cells of the endometrium. Taken together, these results suggest that uterine Enpp2 may be regulated by E2 and plays a role in reproductive functions during female rat development.


Sujet(s)
Animaux , Femelle , Rats , Oestradiol/pharmacologie , Cycle oestral/physiologie , Régulation de l'expression des gènes/physiologie , Immunohistochimie , Mifépristone/pharmacologie , Phosphodiesterases/génétique , Progestérone/pharmacologie , ARN messager/génétique , Rat Sprague-Dawley , Utérus/métabolisme
20.
Biol. Res ; 44(1): 89-105, 2011. ilus, tab
Article de Anglais | LILACS | ID: lil-591869

RÉSUMÉ

Since the discovery of the low-density lipoprotein receptor (LDLR) and its association with familial hypercholesterolemia in the early 1980s, a family of structurally related proteins has been discovered that has apolipoprotein E as a common ligand, and the broad functions of its members have been described. LRP2, or megalin, is a member of the LDLR family and was initially called gp330. Megalin is an endocytic receptor expressed on the apical surface of several epithelial cells that internalizes a variety of ligands including nutrients, hormones and their carrier proteins, signaling molecules, morphogens, and extracellular matrix proteins. Once internalized, these ligands are directed to the lysosomal degradation pathway or transported by transcytosis from one side of the cell to the opposite membrane. The availability of megalin at the cell surface is controlled by several regulatory mechanisms, including the phosphorylation of its cytoplasmic domain by GSK3, the proteolysis of the extracellular domain at the cell surface (shedding), the subsequent intramembrane proteolysis of the transmembrane domain by the gamma-secretase complex, and exosome secretion. Based on the important roles of its ligands and its tissue expression pattern, megalin has been recognized as an important component of many pathological conditions, including diabetic nephropathy, Lowe syndrome, Dent disease, Alzheimer's disease (AD) and gallstone disease. In addition, the expression of megalin and some of its ligands in the central and peripheral nervous system suggests a role for this receptor in neural regeneration processes. Despite its obvious importance, the regulation of megalin expression is poorly understood. In this review, we describe the functions of megalin and its association with certain pathological conditions as well as the current understanding of the mechanisms that underlie the control of megalin expression.


Sujet(s)
Humains , Maladie d'Alzheimer/métabolisme , /physiologie , Maladie d'Alzheimer/physiopathologie , Transport biologique/physiologie , Cholestérol/physiologie , Calculs biliaires/métabolisme , Calculs biliaires/physiopathologie , Régulation de l'expression des gènes/physiologie , Homéostasie/physiologie , Maladies du rein/métabolisme , Maladies du rein/physiopathologie , /génétique , /métabolisme , Distribution tissulaire/physiologie
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