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1.
Compr Rev Food Sci Food Saf ; 20(3): 2699-2715, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33719191

RESUMO

Furan is generally produced during thermal processing of various foods including baked, fried, and roasted food items such as cereal products, coffee, canned, and jarred prepared foods as well as in baby foods. Furan is a toxic and carcinogenic compound to humans and may be a vital hazard to infants and babies. Furan could be formed in foods through thermal degradation of carbohydrates, dissociation of amino acids, and oxidation of polyunsaturated fatty acids. The detection of furan in food products is difficult due to its high volatility and low molecular weight. Headspace solid-phase microextraction coupled with gas chromatography/mass spectrometer (GC/MS) is generally used for analysis of furan in food samples. The risk assessment of furan can be characterized using margin of exposure approach (MOE). Conventional strategies including cooking in open vessels, reheating of commercially processed foods with stirring, and physical removal using vacuum treatment have remained unsuccessful for the removal of furan due to the complex production mechanisms and possible precursors of furan. The innovative food-processing technologies such as high-pressure processing (HPP), high-pressure thermal sterilization (HPTS), and Ohmic heating have been adapted for the reduction of furan levels in baby foods. But in recent years, only HPP has gained interest due to successful reduction of furan because of its nonthermal mechanism. HPP-treated baby food products are commercially available from different food companies. This review summarizes the mechanism involved in the formation of furan in foods, its toxicity, and identification in infant foods and presents a solution for limiting its formation, occurrence, and retention using novel strategies.


Assuntos
Contaminação de Alimentos , Alimentos Infantis , Contaminação de Alimentos/análise , Furanos/análise , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Lactente , Alimentos Infantis/análise , Microextração em Fase Sólida
2.
J Cell Sci ; 126(Pt 2): 445-53, 2013 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-23230143

RESUMO

p27(Kip1), a cyclin-dependent kinase (CDK) inhibitor, is a multi-functional protein that regulates various cellular activities. Trophoblast proliferation, migration and invasion are some of the key processes of placental development. We have recently reported that Nodal, a member of the transforming growth factor-ß (TGF-ß) superfamily, inhibits human trophoblast cell proliferation, migration and invasion. In the present study, we investigated the mechanism by which Nodal regulates trophoblast activities. We found that Nodal increased p27 mRNA and protein levels by enhancing their stability. Interestingly, Nodal signaling also induced nuclear export of p27 and CDK2. Cytoplasmic translocation of p27 induced by Nodal requires p27 phosphorylation at S10. In addition, Nodal enhanced the association of p27 with CDK2, CDK5 and a microtubule-destabilizing protein, stathmin, and induced stathmin phosphorylation at S25 and S38. Furthermore, Nodal increased tubulin stability as revealed by immunofluorescent staining of acetylated tubulin. Finally, silencing of p27 reversed the inhibitory effect of Nodal on trophoblast cell proliferation, migration and invasion. Taken together, our findings revealed a novel function of simultaneous p27 and CDK2 cytoplasmic mislocalization in mediating growth-factor-regulated cell proliferation, migration and invasion.


Assuntos
Quinase 2 Dependente de Ciclina/metabolismo , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Citoplasma/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas de Membrana/metabolismo , Trofoblastos/metabolismo , Processos de Crescimento Celular/fisiologia , Linhagem Celular , Movimento Celular/fisiologia , Quinase 2 Dependente de Ciclina/genética , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Proteínas de Membrana/genética , Fosforilação , Transdução de Sinais , Transfecção , Trofoblastos/citologia , Trofoblastos/enzimologia , Regulação para Cima
3.
Food Chem ; 451: 139465, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-38677132

RESUMO

This work aimed to synthesize oregano essential oil/ß-cyclodextrin microcapsules (OEO/ß-CDs) and then prepare gelatin-based controlled-release antibacterial films with different OEO/ß-CDs contents (0%-2%) for chilling preservation of grass carp fillets. The results of FTIR, XRD, DSC and accelerated release ratio showed that OEO was successfully encapsulated in OEO/ß-CDs and its thermal stability was effectively improved. Moreover, at 2% of addition amount of OEO/ß-CDs, the tensile strength of the films increased from 14.43 MPa to 18.72 MPa. In addition, the films showed significant antibacterial activity against Pseudomonas (61.52%), Aeromonas (62.87%), and Shewanella putrefaciens (66.67%). Preservation experiments showed that the films effectively prevented the increase of TVB-N, and TBA value of the refrigerated fillets and significantly suppressed the growth of spoilage organisms, thus extending the shelf life by 2-3 days. Therefore, the synthesized film has promising potential as an active packaging material for the preservation of grass carp.


Assuntos
Antibacterianos , Cápsulas , Carpas , Preparações de Ação Retardada , Conservação de Alimentos , Gelatina , Óleos Voláteis , Origanum , beta-Ciclodextrinas , Animais , Carpas/microbiologia , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Gelatina/química , Conservação de Alimentos/métodos , Antibacterianos/farmacologia , Antibacterianos/química , Origanum/química , Cápsulas/química , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacologia , beta-Ciclodextrinas/química , Embalagem de Alimentos/instrumentação , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Temperatura Baixa
4.
Mol Biol Rep ; 40(10): 5685-9, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24065524

RESUMO

Prolactin (PRL) is a polypeptide hormone, secreted mainly by the anterior pituitary gland. It is involved in many endocrine activities. The key functions of PRL are related to reproduction and lactation in mammals. To ascertain the presence of polymorphisms in the bovine PRL gene (bPRL), the bPRL gene was sequenced. Five mutations were identified in exonic region and eleven in associated intronic regions in 100 cattle from four Pakistani cattle breeds. Haplotype of predicted amino acid changes represent a common alteration at codon 222 from R-Arginine into K-Lysine in all four breeds. Significant statistical variations were observed in the distribution of single nucleotide polymorphism (SNP) in various cattle populations. However, on basis of present study, an association of these SNPs with milk performance traits in four Pakistani cow breeds cannot be truly replicated but at least can be effective DNA markers for some of the breeds studied. Linkage analysis between these SNPs on larger populations can be useful for the association with milk production traits. Furthermore, present study may be used for marker-assisted selection and management in cattle breeding program in local cattle breeds.


Assuntos
Bovinos/genética , Prolactina/genética , Alelos , Animais , Cruzamento , Éxons/genética , Íntrons/genética , Paquistão , Polimorfismo de Nucleotídeo Único/genética
5.
Chronic Illn ; 19(1): 221-232, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-35040356

RESUMO

OBJECTIVE: This study aimed to translate to Arabic language and culturally adapt the Self-Efficacy for Managing Chronic Diseases Scale (SEMCD) and The Perceived Medical Condition Self-Management Scale (PMCSMS). DESIGN: The translation of the two scales was conducted according to the framework by Beaton et al. Then, these scales were tested in a cross-sectional correlational study with 85 Arabic-speaking participants with multimorbidities in Qatar. Psychometric tests to assess the reliability and validity of the translated scales were done. RESULTS: The internal consistency reliability (α) for the Arabic version of both PMCSMS and SEMCD scales was 0.953 and 0.949, respectively. Correlation coefficient (r) used to assess convergent validity between the PMCSMS and the SEMCD was 0.76 (p < 0.5). Number of morbidities negatively correlated with scores of self-management self- efficacy scores of the PMCSMS and the SEMCD (r = -0.50 and -0.51, respectively). DISCUSSION: The translated PMCSMS and SEMCD are valid and reliable scales that can be used by researchers and healthcare providers to assess levels of self-efficacy among individuals with multimorbidities. The translated scales can be employed in research or interventional programmes that aim to improve self-management or self-efficacy among individuals with multimorbidities.


Assuntos
Comparação Transcultural , Autogestão , Humanos , Autoeficácia , Reprodutibilidade dos Testes , Estudos Transversais , Multimorbidade , Inquéritos e Questionários , Idioma , Psicometria
6.
J Food Sci ; 88(8): 3341-3356, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37421346

RESUMO

During the processing of maize, Stigma maydis, also known as corn silk, is normally discarded as waste. Phytochemical research was carried out on the S. maydis to use it as a valuable source of bioactive components. This research aimed to maximize the recovery of free and bound phenolic compounds from corn silk under optimal experimental conditions. Response surface design was operated to optimize the alkaline hydrolysis extraction of bound phytochemicals from corn silk based on total phenolic content and DPPH radical scavenging activity. The optimum conditions (i.e., NaOH concentration 2 M, digestion time 135 min, digestion temperature of 37.5°C, the solid-to-solvent ratio of 1:17.5, and acetone) were obtained. The optimum parameters were used to extract the corn silk. The structures of two compounds isolated from ethyl acetate extracts were then identified as friedelin (1) and (E)-4-(4-hydroxy-3-methoxyphenyl) but-3-en-2-one (2). The DPPH, H2 O2 , and ABTS % inhibition of the compounds is as follows: compound (1) 74.81%, 76.8%, 70.33% and compound (2) 70.37%, 56.70% and 57.46%, respectively. The current study has opened previously unexplored perspectives of the composition of bound compounds in corn silk and established the foundations for more effective processing and utilization of corn waste. PRACTICAL APPLICATION: Bound phenolic compounds from corn silk under optimal experimental conditions were obtained. Corn silk can be utilized as a type of medicinal herb as well as a source of inexpensive natural antioxidants.


Assuntos
Antioxidantes , Plantas Medicinais , Antioxidantes/química , Extratos Vegetais/química , Zea mays/química , Fenóis/química , Seda
7.
Polymers (Basel) ; 15(24)2023 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-38139899

RESUMO

In the current era, the treatment of collagen hydrogels with natural phenolics for the improvement in physicochemical properties has been the subject of considerable attention. The present research aimed to fabricate collagen hydrogels cross-linked with gallic acid (GA) and ellagic acid (EA) at different concentrations depending on the collagen dry weight. The structural, enzymatic, thermal, morphological, and physical properties of the native collagen hydrogels were compared with those of the GA/EA cross-linked hydrogels. XRD and FTIR spectroscopic analyses confirmed the structural stability and reliability of the collagen after treatment with either GA or EA. The cross-linking also significantly contributed to the improvement in the storage modulus, of 435 Pa for 100% GA cross-linked hydrogels. The thermal stability was improved, as the highest residual weight of 43.8% was obtained for the hydrogels cross-linked with 50% GA in comparison with all the other hydrogels. The hydrogels immersed in 30%, 50%, and 100% concentrations of GA also showed improved swelling behavior and porosity, and the highest resistance to type 1 collagenase (76.56%), was obtained for 50% GA cross-linked collagen hydrogels. Moreover, GA 100% and EA 100% obtained the highest denaturation temperatures (Td) of 74.96 °C and 75.78 °C, respectively. In addition, SEM analysis was also carried out to check the surface morphology of the pristine collagen hydrogels and the cross-linked collagen hydrogels. The result showed that the hydrogels cross-linked with GA/EA were denser and more compact. However, the improved physicochemical properties were probably due to the formation of hydrogen bonds between the phenolic hydroxyl groups of GA and EA and the nitrogen atoms of the collagen backbone. The presence of inter- and intramolecular cross-links between collagen and GA or EA components and an increased density of intermolecular bonds suggest potential hydrogen bonding or hydrophobic interactions. Overall, the present study paves the way for further investigations in the field by providing valuable insights into the GA/EA interaction with collagen molecules.

8.
Am J Pathol ; 178(3): 1177-89, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21356369

RESUMO

Trophoblast cell invasion into the uterus is an essential process for successful pregnancy, and shallow invasion of trophoblasts into the maternal decidua is linked to preeclampsia. We have reported that Nodal, a member of the transforming growth factor-ß superfamily, acts through activin receptor-like kinase 7 (ALK7) to inhibit trophoblast proliferation and to induce apoptosis. In this study, we examined the spatial and temporal expression patterns of Nodal and ALK7 in human placenta from normal and preeclamptic pregnancies and investigated whether Nodal regulated trophoblast migration and invasion. Nodal and ALK7 were detected in villous and extravillous trophoblast cell populations in early gestation, and their levels were strongly up-regulated in preeclamptic placenta. Overexpression of Nodal or constitutively active ALK7 decreased cell migration and invasion, whereas knockdown of Nodal and ALK7 had the opposite effects. In placental explant culture, treatment with Nodal inhibited trophoblast outgrowth, whereas Nodal small-interfering RNA strongly induced the expansion of explants and the migration of extravillous trophoblast cells. Nodal stimulated the secretion of tissue inhibitor of metalloproteinase-1 and inhibited matrix metalloproteinase (MMP)-2 and MMP-9 activity. These findings suggest that the Nodal/ALK7 pathway plays important roles in human placentation and that its abnormal signaling may contribute to the development of preeclampsia.


Assuntos
Receptores de Ativinas Tipo I/metabolismo , Movimento Celular , Proteína Nodal/metabolismo , Pré-Eclâmpsia/etiologia , Transdução de Sinais , Trofoblastos/enzimologia , Trofoblastos/patologia , Feminino , Humanos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Placenta/enzimologia , Placenta/patologia , Placentação , Pré-Eclâmpsia/enzimologia , Pré-Eclâmpsia/metabolismo , Gravidez , RNA Interferente Pequeno/metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismo
9.
Food Chem ; 360: 130068, 2021 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-34029925

RESUMO

Collagen Type I derived from fish is mainly limited by its poor physicochemical properties for further applications. In this study, we developed epigallocatechin gallate (EGCG) cross-linked collagen hydrogels (EC hydrogels) to realize physicochemical improvements, basing on the interaction mechanism between collagen and EGCG. The integrity of collagen framework with slight secondary structure change in the presence of EGCG was confirmed. The stronger stability of collagen fibrils was proved by slower swelling ratio, declined enzymatic degradation, improved thermal analysis and mechanical test due to EGCG modification. To illustrate the potential mechanism between collagen and EGCG, molecular docking was used to identify both covalent (CN bond, between lysine of collagen and C2-ring B of EGCG) and non-covalent bonds (hydrogen bond and hydrophobic interaction) within in EC hydrogel. Taken together, this work would offer some insights into the further study about the interaction between EGCG and collagen.


Assuntos
Catequina/análogos & derivados , Colágeno/química , Hidrogéis/química , Animais , Catequina/química , Catequina/metabolismo , Colágeno/metabolismo , Hidrogéis/metabolismo , Ligação de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Reologia , Eletricidade Estática , Temperatura de Transição
10.
Food Chem ; 345: 128655, 2021 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-33302098

RESUMO

The aim of this study was to determine the influence and mechanism of combining EGCG with TGase on properties of myofibrillar protein (MP) gel. A double-crosslinked effect was observed when EGCG and TGase were added into MP gel. Breaking force, deformation, water holding capacity and hardness of double-crosslinked MP gel increased by 25.3 ± 3.0 g, 0.5 ± 0.3 mm, 1.76 ± 0.4% and 34.11 ± 2.56 g, compared with those of TGase induced gel. Light microscopy and low-field nuclear magnetic resonance results indicated with EGCG content increasing, pores and structure of double-crosslinked gels became smaller and denser, T22 decreased from 266.162 ms to 252.845 ms and its proportion increased from 94.103% to 96.956%. Molecular docking illustrated covalent and non-covalent interactions between EGCG and myosin heavy chain Ⅱ A, and confirmed TGase catalytic mechanism with myosin heavy chain Ⅱ A as substrate. Therefore the mixture of EGCG and TGase could be used as novel cross-linker in surimi.


Assuntos
Biocatálise , Catequina/análogos & derivados , Fenômenos Químicos , Simulação de Acoplamento Molecular , Proteínas Musculares/química , Transglutaminases/metabolismo , Animais , Catequina/química , Géis , Proteínas Musculares/metabolismo
11.
Polymers (Basel) ; 12(10)2020 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-33027939

RESUMO

The effects of green tea extract (GTE) at acidic and alkaline pH (pH 3 and 11, respectively) on the physico-mechanical, thermal, and water transmission properties of silver carp surimi-based edible films were investigated. Incorporation of GTE significantly (p < 0.05) reduced elongation at break (EAB) but improved tensile strength (TS). Significant (p < 0.05) decreases in the solubility of films and water vapor permeability (WVP) were also perceived in GTE-containing films. Transparency and color were also affected, depending upon the concentration and pH. Films prepared at an acidic pH demonstrated significantly (p < 0.05) improved WVP, transparency, solubility, and thermal stability compared to those prepared at an alkaline pH. The protein pattern of films revealed a strong interaction between phenolic compounds of GTE and surimi proteins as evidenced by the presence of a myosin heavy chain (MCH) at the top of the gel. Generally, the addition of GTE at an acidic pH had significantly (p < 0.05) greater impact on film properties than at an alkaline pH and could offer great potential for surimi protein-based edible films with improved properties.

12.
J Public Health Res ; 9(4): 1716, 2020 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-33117756

RESUMO

Background: The current nursing workforce needs to be skilled, confident, and competent to address the rapid change in Primary Health Care services, to align with the National Vision of Qatar. This is emphasized in the Primary Health Care Strategy 2018-2023. This strategy outlines the need of having a skilled nursing workforce to administer and educate the public about the importance of immunizations. Primary Health Care nurses emanate from several nationalities and hence possess various level of knowledge and background related to immunization administration. Design and methods: To assess Primary Health Care nurses' knowledge before and after the delivered immunization education program in Qatar. The study was part of a Sequential mixed method research study that aimed to assess the Primary Health Care nurses' knowledge, attitude, and practice of nurses before and after the conducted training program. A self-administered survey, specifically developed for the research project was loaded in Qualtrics and sent to participants of the immunization education program before and after training. Participants were trained in two cohorts from October to December, 2018 in Qatar. For the purposes of this paper, data about the immunization knowledge was statistically analyzed using the SPSS Software version 25 and Microsoft Excel. Results: Specific areas were identified as knowledge gaps among the participants of the two cohorts. Moreover, the developed education program showed overall improvement in the participants' knowledge. Conclusions: The study results have demonstrated that the delivered immunization education program significantly increases the participants' knowledge about immunization in certain areas of the primary healthcare clinics they work, including vaccine safety, efficiency, and contraindications in the delivery of vaccines to the public.

13.
Front Microbiol ; 11: 1854, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32849446

RESUMO

The coronavirus disease 2019 (COVID-19) is a clinical syndrome caused by severe acute respiratory syndrome corona virus-2 (SARS-CoV-2). COVID-19 was declared a pandemic by the World Health Organization (WHO) on March 11, 2020 due to its rapid and extensive spread among many countries through its very contagious nature and its high mortality among the elderly and infirm. Recently, data on the survival of SARS-CoV-2 on contact surfaces has been reported, but there is none on the survival of COVID-19 on food surfaces and packages. The potential survival and transmission of SARS-CoV-2 on/via food and packages are discussed based on data available for other respiratory viruses such as SARS-CoV and MERS-CoV. However, studies are needed to explore its transmission via food and survival on food packaging materials. The implementation of food safety management systems such as Hazard Analysis and Critical Control Points (HACCP), and Good Manufacturing Practices (GMP) are important to reduce the risk of COVID-19 infection. Cleaning, sanitation, good hygienic practices, and active packaging are also needed from farm to fork.

14.
Food Funct ; 11(11): 9560-9572, 2020 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-33146213

RESUMO

The rheological behavior, assembly measurements, thermal stability, molecular conformation, and molecular interactions of myofibrillar proteins (MP) modified by transglutaminase (TGase) and epigallocatechin-3-gallate (EGCG) were investigated. Fluorescence and ultraviolet spectra showed that TGase and ECCG quenched the endogenous fluorescence of MP and improved the exposure of chromophoric and hydrophobic groups of MP caused by unraveling the tertiary structure. TGase and EGCG treatment increased the α-helix and ß-turn contents and the structure became more ordered. It also increased the thermal stability of MP and the storage modulus of MP gels as reflected in the rheological and thermal properties tests. Meanwhile, changes in SDS-PAGE, FT-IR, and sulfhydryl contents showed that TGase increased the disulfide bond contents, whereas it decreased after EGCG was added, suggesting that EGCG could react with MP via non-covalent and covalent interactions. Assembly measurements illustrated that TGase increased the turbidity and particle size of MP, while TGase and EGCG treatment accelerated MP aggregation and it was associated with the increase in the EGCG concentration, which was further confirmed by SEM and AFM. This work could contribute new insights into the synergistic effect of TGase and EGCG on modifying MP and guide for its application in surimi processing.


Assuntos
Catequina/análogos & derivados , Alimento Funcional , Miofibrilas/química , Transglutaminases/química , Animais , Catequina/química , Interações Medicamentosas , Peixes , Humanos , Reologia
15.
Plant Physiol Biochem ; 102: 167-79, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26949025

RESUMO

Calcium (Ca(2+)) has emerged as a significant secondary messenger that regulates the activities of hormonal and environmental signals that are associated with biotic and abiotic stresses. Ca(2+) binding proteins typically contain a Ca(2+) binding EF-hand (a helix-loop-helix structure) motif. In this study, tomato genes encoding calmodulin-like (CML) proteins that possess EF-hand motifs and no other identifiable functional domains were analyzed. Using genome analysis and BLAST searches in database, 52 CML genes were identified in tomato. Comprehensive analyses, including evolutionary relationships, gene structures, chromosomal locations, functional annotations, and gene duplications, were performed. Distribution mapping exhibited that 52 SlCML proteins containing different intron/exon patterns were unevenly distributed among ten chromosomes. In addition, 24 SlCML proteins were predicted as segmentally duplicated. Conserved motifs, promoter cis-regulatory elements, organ-based expression patterns and expression analyses indicated the potential responsiveness of SlCML proteins to abiotic stresses and phytohormones. These results illustrate the complexity of the CML gene family and indicate a potential vital role for these molecules in tomato growth and development as Ca(2+) signal transducers.


Assuntos
Calmodulina , Cromossomos de Plantas/genética , Regulação da Expressão Gênica/fisiologia , Família Multigênica , Proteínas de Plantas , Solanum lycopersicum , Calmodulina/biossíntese , Calmodulina/genética , Bases de Dados de Ácidos Nucleicos , Duplicação Gênica , Estudo de Associação Genômica Ampla , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética
16.
J Clin Endocrinol Metab ; 89(11): 5523-34, 2004 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-15531507

RESUMO

Human epithelial ovarian cancer is the most lethal female cancer. Hormones and growth factors, including the TGF-beta superfamily, have been suggested to play a role in ovarian tumorigenesis. The biological effects of TGF-beta superfamily are mediated by type I and type II serine/threonine kinase receptors and by intracellular Smad proteins. Recently, we have cloned four transcripts of human activin receptor-like kinase 7 (ALK7), a type I receptor for Nodal. In this study, we have investigated the role of Nodal and ALK7 in four ovarian cancer cell lines, OV2008, C13*, A2780-s, and A2780-cp. Overexpression of Nodal resulted in a significant decrease in the number of metabolically active cells. This effect was mimicked by a constitutively active ALK7 (ALK7-ca) but blocked by dominant negative mutants of ALK7, Smad2, or Smad3. Transient transfection of Nodal and ALK7-ca significantly decreased X-linked inhibitor of apoptosis protein (Xiap) expression, activated both caspase-3 and caspase-9, and increased apoptosis as determined by Hoechst nuclear staining and flow cytometry. In addition, Nodal and ALK7-ca also inhibited cell proliferation as measured by 5-bromo-2'-deoxyuridine (BrdU) assays. Interestingly, the effects of Nodal and ALK7-ca were more potent in chemosensitive A2780-s cells than in its chemoresistant counterpart, A2780-cp cells. These findings demonstrate that Nodal induces apoptosis and inhibits proliferation via ALK7 and Smad2/3 and that the effect of Nodal-ALK7 on apoptosis may be mediated in part by the down-regulation of Xiap and activation of caspase-9 and caspase-3.


Assuntos
Receptores de Ativinas Tipo I/fisiologia , Apoptose , Neoplasias Ovarianas/patologia , Fator de Crescimento Transformador beta/fisiologia , Caspases/fisiologia , Divisão Celular , Linhagem Celular Tumoral , Proteínas de Ligação a DNA/fisiologia , Feminino , Humanos , Proteína Nodal , Proteínas/fisiologia , Transdução de Sinais , Proteína Smad2 , Proteína Smad3 , Transativadores/fisiologia , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo X
17.
J Biol Chem ; 279(30): 31277-86, 2004 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-15150278

RESUMO

Nodal, a member of the transforming growth factor-beta superfamily, is known to play critical roles in early vertebrate development, but its functions in extraembryonic tissues are unclear. ALK7 is a type I receptor for Nodal. Recently, we demonstrated that Nodal mRNA and several ALK7 transcripts are expressed in human placenta throughout pregnancy (Roberts, H. J., Hu, S., Qiu, Q., Leung, P. C. K., Cannigia, I., Gruslin, A., Tsang, B., and Peng, C. (2003) Biol. Reprod. 68, 1719-1726). In this study, we determined the role of Nodal and ALK7 in trophoblast cell proliferation and apoptosis. Overexpression of Nodal in normal trophoblast cells (HTR8/SVneo) and several choriocarcinoma cell lines resulted in a significant decrease in the number of metabolically active cells. The effect of Nodal could be mimicked by constitutively active ALK7 (ALK7-ca), but was blocked by kinase-deficient ALK7. The growth inhibitory effect of Nodal was also blocked by dominant-negative Smad2/3. Overexpression of Nodal and ALK7-ca induced apoptosis in trophoblast cells as determined by Hoechst staining, flow cytometry, and caspase-3 Western blotting. In addition, Nodal and ALK7-ca decreased the number of proliferating cells as measured by bromodeoxyuridine assays. Furthermore, overexpression of Nodal or ALK7-ca increased p27 expression, but reduced the levels of Cdk2 and cyclin D(1). Taken together, this study demonstrates for the first time that Nodal, acting through ALK7 and Smad2/3, inhibits proliferation and induces apoptosis in human trophoblast cells. Our findings also suggest that the Nodal-ALK7 pathway inhibits cell proliferation by inducing G(1) cell cycle arrest and that this effect is mediated in part by the p27-cyclin E/Cdk2 pathway.


Assuntos
Receptores de Ativinas Tipo I/fisiologia , Fator de Crescimento Transformador beta/fisiologia , Trofoblastos/citologia , Receptores de Ativinas Tipo I/genética , Apoptose/fisiologia , Sequência de Bases , Divisão Celular/fisiologia , Linhagem Celular , Primers do DNA/genética , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/fisiologia , Feminino , Fase G1 , Expressão Gênica , Humanos , Proteína Nodal , Gravidez , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais , Proteína Smad2 , Proteína Smad3 , Transativadores/genética , Transativadores/fisiologia , Transfecção , Fator de Crescimento Transformador beta/genética , Trofoblastos/fisiologia
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