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1.
Nat Commun ; 13(1): 3933, 2022 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-35798722

RESUMO

Cassava mosaic disease (CMD) suppresses cassava yields across the tropics. The dominant CMD2 locus confers resistance to cassava mosaic geminiviruses. It has been reported that CMD2-type landraces lose resistance after regeneration through de novo morphogenesis. As full genome bisulfite sequencing failed to uncover an epigenetic mechanism for this loss of resistance, whole genome sequencing and genetic variant analysis was performed and the CMD2 locus was fine-mapped to a 190 kilobase interval. Collectively, these data indicate that CMD2-type resistance is caused by a nonsynonymous, single nucleotide polymorphism in DNA polymerase δ subunit 1 (MePOLD1) located within this region. Virus-induced gene silencing of MePOLD1 in a CMD-susceptible cassava variety produced a recovery phenotype typical of CMD2-type resistance. Analysis of other CMD2-type cassava varieties identified additional candidate resistance alleles within MePOLD1. Genetic variation of MePOLD1, therefore, could represent an important genetic resource for resistance breeding and/or genome editing, and elucidating mechanisms of resistance to geminiviruses.


Assuntos
Begomovirus , Geminiviridae , Manihot , DNA Polimerase III/genética , Resistência à Doença/genética , Geminiviridae/genética , Manihot/genética , Mutação , Melhoramento Vegetal , Doenças das Plantas/genética
2.
Multimed Tools Appl ; 81(19): 27631-27655, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35368858

RESUMO

COVID-19 is a viral disease that in the form of a pandemic has spread in the entire world, causing a severe impact on people's well being. In fighting against this deadly disease, a pivotal step can prove to be an effective screening and diagnosing step to treat infected patients. This can be made possible through the use of chest X-ray images. Early detection using the chest X-ray images can prove to be a key solution in fighting COVID-19. Many computer-aided diagnostic (CAD) techniques have sprung up to aid radiologists and provide them a secondary suggestion for the same. In this study, we have proposed the notion of Pearson Correlation Coefficient (PCC) along with variance thresholding to optimally reduce the feature space of extracted features from the conventional deep learning architectures, ResNet152 and GoogLeNet. Further, these features are classified using machine learning (ML) predictive classifiers for multi-class classification among COVID-19, Pneumonia and Normal. The proposed model is validated and tested on publicly available COVID-19 and Pneumonia and Normal dataset containing an extensive set of 768 images of COVID-19 with 5216 training images of Pneumonia and Normal patients. Experimental results reveal that the proposed model outperforms other previous related works. While the achieved results are encouraging, further analysis on the COVID-19 images can prove to be more reliable for effective classification.

3.
Phys Eng Sci Med ; 44(4): 1257-1271, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34609703

RESUMO

According to the World Health Organization (WHO), novel coronavirus (COVID-19) is an infectious disease and has a significant social and economic impact. The main challenge in fighting against this disease is its scale. Due to the outbreak, medical facilities are under pressure due to case numbers. A quick diagnosis system is required to address these challenges. To this end, a stochastic deep learning model is proposed. The main idea is to constrain the deep-representations over a Gaussian prior to reinforce the discriminability in feature space. The model can work on chest X-ray or CT-scan images. It provides a fast diagnosis of COVID-19 and can scale seamlessly. The work presents a comprehensive evaluation of previously proposed approaches for X-ray based disease diagnosis. The approach works by learning a latent space over X-ray image distribution from the ensemble of state-of-the-art convolutional-nets, and then linearly regressing the predictions from an ensemble of classifiers which take the latent vector as input. We experimented with publicly available datasets having three classes: COVID-19, normal and pneumonia yielding an overall accuracy and AUC of 0.91 and 0.97, respectively. Moreover, for robust evaluation, experiments were performed on a large chest X-ray dataset to classify among Atelectasis, Effusion, Infiltration, Nodule, and Pneumonia classes. The results demonstrate that the proposed model has better understanding of the X-ray images which make the network more generic to be later used with other domains of medical image analysis.


Assuntos
COVID-19 , Aprendizado Profundo , Algoritmos , Humanos , Redes Neurais de Computação , SARS-CoV-2 , Raios X
4.
Plant Biotechnol J ; 19(4): 844-854, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33190345

RESUMO

Delivering the benefits of agricultural biotechnology to smallholder farmers requires that resources be directed towards staple food crops. To achieve effect at scale, beneficial traits must be integrated into multiple, elite farmer-preferred varieties with relevance across geographical regions. The staple root crop cassava (Manihot esculenta) is consumed for dietary calories by more than 800 million people, but its tuberous roots provide insufficient iron and zinc to meet nutritional needs. In Africa, cassava yields are furthermore limited by the virus diseases, cassava mosaic disease (CMD) and cassava brown streak disease (CBSD). In this study, we strove to develop cassava displaying high-level resistance to CBSD and CMD to attain food and economic security for cassava farmers, along with biofortified levels of iron and zinc to enhance consumer health. RNAi-mediated technology was used to achieve resistance to CBSD in two East African and one Nigerian farmer-preferred cultivars that harboured resistance to CMD. The Nigerian cvs. TMS 95/0505 and TMS 91/02324 were modified with T-DNA imparting resistance to CBSD, along with AtIRT1 (major iron transporter) and AtFER1 (ferritin) transgenes to achieve nutritionally significant levels of iron and zinc in cassava storage roots (145 and 40 µg/g dry weight, respectively). The inherent resistance to CMD was maintained in all four disease resistant and mineral enhanced cassava cultivars described here, demonstrating that this technique could be deployed across multiple farmer-preferred varieties to benefit the food and nutritional security of consumers in Africa.


Assuntos
Manihot , África , Biofortificação , Resistência à Doença/genética , Humanos , Manihot/genética , Minerais , Doenças das Plantas
5.
Preprint em Inglês | medRxiv | ID: ppmedrxiv-20063461

RESUMO

According to the World Health Organization (WHO), the coronavirus (COVID-19) pandemic is putting even the best healthcare systems across the world under tremendous pressure. The early detection of this type of virus will help in relieving the pressure of the healthcare systems. Chest X-rays has been playing a crucial role in the diagnosis of diseases like Pneumonia. As COVID-19 is a type of influenza, it is possible to diagnose using this imaging technique. With rapid development in the area of Machine Learning (ML) and Deep learning, there had been intelligent systems to classify between Pneumonia and Normal patients. This paper proposes the machine learning-based classification of the extracted deep feature using ResNet152 with COVID-19 and Pneumonia patients on chest X-ray images. SMOTE is used for balancing the imbalanced data points of COVID-19 and Normal patients. This non-invasive and early prediction of novel coronavirus (COVID-19) by analyzing chest X-rays can further be used to predict the spread of the virus in asymptomatic patients. The model is achieving an accuracy of 0.973 on Random Forest and 0.977 using XGBoost predictive classifiers. The establishment of such an approach will be useful to predict the outbreak early, which in turn can aid to control it effectively.

6.
Phytother Res ; 33(5): 1457-1468, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30838706

RESUMO

A double-blind, placebo-controlled human trial was conducted to evaluate the safety and efficacy of a standardized oral supplementation of Boswellin®, a novel extract of Boswellia serrata extract (BSE) containing 3-acetyl-11-keto-ß-boswellic acid (AKBBA) with ß-boswellic acid (BBA). A total of 48 patients with osteoarthritis (OA) of the knee were randomized and allocated to the BSE and placebo groups for intervention. Patients were administered BSE or placebo for a period of 120 days. The trial results revealed that BSE treatment significantly improved the physical function of the patients by reducing pain and stiffness compared with placebo. Radiographic assessments showed improved knee joint gap and reduced osteophytes (spur) confirming the efficacy of BSE treatment. BSE also significantly reduced the serum levels of high-sensitive C-reactive protein, a potential inflammatory marker associated with OA of the knee. No serious adverse events were reported. This is the first study with BSE conducted for a period of 120 days, longer than any other previous clinical trial on patients with OA of the knee. The findings provide evidence that biologically active constituents of BSE, namely, AKBBA and BBA, act synergistically to exert anti-inflammatory/anti-arthritic activity showing improvement in physical and functional ability and reducing the pain and stiffness.


Assuntos
Boswellia/química , Osteoartrite do Joelho/tratamento farmacológico , Dor/tratamento farmacológico , Extratos Vegetais/uso terapêutico , Idoso , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Medição da Dor , Projetos Piloto , Extratos Vegetais/efeitos adversos , Triterpenos/administração & dosagem
7.
Nat Biotechnol ; 37(3): 323, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30787468

RESUMO

In the version of this article initially published, a relevant work was not cited. The following sentence has been inserted following the sentence ending "Aspergillus phytase" in the third paragraph of the article: "Overexpression of AtIRT1, AtNAS1 and bean FERRITIN in rice resulted in 3.8-fold higher iron and 1.8-fold higher zinc concentrations than in the wild-type control12." A corresponding reference has been added: 12. Boonyaves, K., Wu, T. Y., Gruissem, W. & Bhullar, N. K. Enhanced grain iron levels in rice expressing an IRON-REGULATED METAL TRANSPORTER, NICOTIANAMINE SYNTHASE, and FERRITIN gene cassette. Front. Plant Sci. 8, 130 (2017). The error has been corrected in the HTML and PDF versions of the article.

8.
Nat Biotechnol ; 37(2): 144-151, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30692693

RESUMO

Less than 10% of the estimated average requirement (EAR) for iron and zinc is provided by consumption of storage roots of the staple crop cassava (Manihot esculenta Crantz) in West African human populations. We used genetic engineering to improve mineral micronutrient concentrations in cassava. Overexpression of the Arabidopsis thaliana vacuolar iron transporter VIT1 in cassava accumulated three- to seven-times-higher levels of iron in transgenic storage roots than nontransgenic controls in confined field trials in Puerto Rico. Plants engineered to coexpress a mutated A. thaliana iron transporter (IRT1) and A. thaliana ferritin (FER1) accumulated iron levels 7-18 times higher and zinc levels 3-10 times higher than those in nontransgenic controls in the field. Growth parameters and storage-root yields were unaffected by transgenic fortification in our field data. Measures of retention and bioaccessibility of iron and zinc in processed transgenic cassava indicated that IRT1 + FER1 plants could provide 40-50% of the EAR for iron and 60-70% of the EAR for zinc in 1- to 6-year-old children and nonlactating, nonpregnant West African women.


Assuntos
Biofortificação , Ferritinas/química , Engenharia Genética/métodos , Ferro/química , Manihot/genética , África Ocidental , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Transporte de Cátions/genética , Ferritinas/genética , Mutação , Valor Nutritivo , Fenótipo , Raízes de Plantas , Plantas Geneticamente Modificadas , Zinco
9.
Plant Biotechnol J ; 16(6): 1186-1200, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29193665

RESUMO

Storage roots of cassava (Manihot esculenta Crantz), a major subsistence crop of sub-Saharan Africa, are calorie rich but deficient in essential micronutrients, including provitamin A ß-carotene. In this study, ß-carotene concentrations in cassava storage roots were enhanced by co-expression of transgenes for deoxy-d-xylulose-5-phosphate synthase (DXS) and bacterial phytoene synthase (crtB), mediated by the patatin-type 1 promoter. Storage roots harvested from field-grown plants accumulated carotenoids to ≤50 µg/g DW, 15- to 20-fold increases relative to roots from nontransgenic plants. Approximately 85%-90% of these carotenoids accumulated as all-trans-ß-carotene, the most nutritionally efficacious carotenoid. ß-Carotene-accumulating storage roots displayed delayed onset of postharvest physiological deterioration, a major constraint limiting utilization of cassava products. Large metabolite changes were detected in ß-carotene-enhanced storage roots. Most significantly, an inverse correlation was observed between ß-carotene and dry matter content, with reductions of 50%-60% of dry matter content in the highest carotenoid-accumulating storage roots of different cultivars. Further analysis confirmed a concomitant reduction in starch content and increased levels of total fatty acids, triacylglycerols, soluble sugars and abscisic acid. Potato engineered to co-express DXS and crtB displayed a similar correlation between ß-carotene accumulation, reduced dry matter and starch content and elevated oil and soluble sugars in tubers. Transcriptome analyses revealed a reduced expression of genes involved in starch biosynthesis including ADP-glucose pyrophosphorylase genes in transgenic, carotene-accumulating cassava roots relative to nontransgenic roots. These findings highlight unintended metabolic consequences of provitamin A biofortification of starch-rich organs and point to strategies for redirecting metabolic flux to restore starch production.


Assuntos
Biofortificação , Metabolismo dos Carboidratos , Carotenoides/metabolismo , Manihot/química , Raízes de Plantas/química , Ácido Abscísico/metabolismo , Armazenamento de Alimentos , Geranil-Geranildifosfato Geranil-Geraniltransferase/genética , Manihot/genética , Manihot/metabolismo , Plantas Geneticamente Modificadas , Solanum tuberosum/química , Amido/biossíntese , Transferases/genética
10.
Int J Oncol ; 50(1): 337, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27779652

RESUMO

Following the publication of this article, which was concerned with the expression of phosphorylated epidermal growth factor receptor (pEGFR) and cyclin D1 activation independently of the expression levels of cyclo-oxygenase-2, an interested reader drew to our attention apparent anomalies associated with the western blot data shown in Fig. 2C. Following an internal investigation at the New York University School of Medicine, we were requested to produce the original film, or the scan of the image of the film, for verification. Unfortunately, we were unable to provide the original film or scanned image to disprove the allegation, since the original pEGFR image could not be found. Therefore, the Investigation Committee recommended that this article be retracted, and we are withdrawing the article in line with the request. All the authors agree to the retraction of this paper. We sincerely regret any inconvenience this has caused. [the original article was published in the International Journal of Oncology 40: 13-20, 2012; DOI: 10.3892/ijo.2011.1211].

11.
ACS Appl Mater Interfaces ; 8(23): 14836-44, 2016 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-27219483

RESUMO

Three-dimensional, functional, and porous scaffolds can find applications in a variety of fields. Here we report the synthesis of hierarchical and interconnected porous sponges using a simple freeze-drying technique, employing collagen extracted from animal skin wastes and superparamagnetic iron oxide nanoparticles. The ultralightweight, high-surface-area sponges exhibit excellent mechanical stability and enhanced absorption of organic contaminants such as oils and dye molecules. Additionally, these biocomposite sponges display significant cellular biocompatibility, which opens new prospects in biomedical uses. The approach highlights innovative ways of transforming biowastes into advanced hybrid materials using simple and scalable synthesis techniques.


Assuntos
Colágeno/química , Recuperação e Remediação Ambiental/instrumentação , Animais , Liofilização , Nanopartículas , Porosidade
12.
Plant Sci ; 240: 170-81, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26475197

RESUMO

Iron is extremely abundant in the soil, but its uptake in plants is limited due to low solubility in neutral or alkaline soils. Plants can rely on rhizosphere acidification to increase iron solubility. AtVIT1 was previously found to be involved in mediating vacuolar sequestration of iron, which indicates a potential application for iron biofortification in crop plants. Here, we have overexpressed AtVIT1 in the starchy root crop cassava using a patatin promoter. Under greenhouse conditions, iron levels in mature cassava storage roots showed 3-4 times higher values when compared with wild-type plants. Significantly, the expression of AtVIT1 showed a positive correlation with the increase in iron concentration of storage roots. Conversely, young leaves of AtVIT1 transgenic plants exhibit characteristics of iron deficiency such as interveinal chlorosis of leaves (yellowing) and lower iron concentration when compared with the wild type plants. Interestingly, the AtVIT1 transgenic plants showed 4 and 16 times higher values of iron concentration in the young stem and stem base tissues, respectively. AtVIT1 transgenic plants also showed 2-4 times higher values of iron content when compared with wild-type plants, with altered partitioning of iron between source and sink tissues. These results demonstrate vacuolar iron sequestration as a viable transgenic strategy to biofortify crops and to help eliminate micronutrient malnutrition in at-risk human populations.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Proteínas de Transporte de Cátions/genética , Ferro/metabolismo , Manihot/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Transporte de Cátions/metabolismo , Manihot/genética , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Caules de Planta/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo
13.
Cancer Med ; 4(11): 1767-80, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26408414

RESUMO

Melinjo (Gnetum gnemon L.) seed extract (MSE) and its active ingredient gnetin C (GC), a resveratrol dimer, have been shown to possess a broad spectrum of pharmacological activities. In this study, we investigated the antitumor activity of MSE and GC using human and murine tumor cell culture models in vitro. The antitumor activity of GC was compared with trans-resveratrol (tRV), a stilbenoid polyphenol. Our results show that MSE and GC at clinically achievable concentrations significantly inhibited the proliferation of pancreatic, prostate, breast, and colon cancer cell types (P < 0.05), without affecting normal cells. Interestingly, GC exerts enhanced antitumor activity than that of tRV (P < 0.05). MSE and GC significantly induced apoptosis in all the cancer cells, indicating MSE and GC inhibit tumor cell growth by inducing apoptosis (P < 0.001). Our findings provide evidence that MSE might induce apoptosis in cancer cells via caspase-3/7-dependent and -independent mechanisms. However, GC might trigger both early and late stage apoptosis in cancer cells, at least in part by activating caspase 3/7-dependent mechanisms. Furthermore, the antitumor efficacy of MSE observed in vitro was also validated in a widely used colon-26 tumor-bearing mouse model. Oral administration of MSE at 50 and 100 mg/kg per day significantly inhibited tumor growth, intratumoral angiogenesis, and liver metastases in BALB/c mice bearing colon-26 tumors (P < 0.05). In conclusion, our findings provide evidence that MSE and GC have potent antitumor activity. Most importantly, we provide the first evidence that MSE inhibits tumor growth, intratumoral angiogenesis, and liver metastasis in a colon-26 tumor-bearing mice.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias do Colo/patologia , Gnetum/química , Extratos Vegetais/farmacologia , Sementes/química , Animais , Antineoplásicos Fitogênicos/química , Apoptose/efeitos dos fármacos , Caspases/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/secundário , Camundongos , Extratos Vegetais/química , Ensaios Antitumorais Modelo de Xenoenxerto
14.
Biochem Biophys Res Commun ; 463(3): 161-6, 2015 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-26003731

RESUMO

Dysregulation of PI3K-AKT-mTOR pathway has been reported in various pathologies, such as cancer and insulin resistance. The proline-rich AKT substrate of 40-kDa (PRAS40), also known as AKT substrate 1 (AKT1S1), lies at the crossroads of these cascades and inhibits the activity of the mTOR complex 1 (mTORC1) kinase. This review discusses the role of PRAS40 and possible feedback mechanisms, and alterations in AKT/PRAS40/mTOR signaling that have been implicated in the pathogenesis of tumor progression. Additionally, we probed new datasets extracted from Oncomine, a cancer microarray database containing datasets derived from patient samples, to further understand the role of PRAS40 (AKT1S1). These data strongly supports the hypothesis that PRAS40 may serve as a potential therapeutic target for various cancers.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Carcinogênese/metabolismo , Neoplasias/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Carcinogênese/efeitos dos fármacos , Carcinogênese/genética , Descoberta de Drogas , Regulação Neoplásica da Expressão Gênica , Humanos , Terapia de Alvo Molecular , Neoplasias/tratamento farmacológico , Neoplasias/genética , Fosfatidilinositol 3-Quinases/metabolismo
15.
Bioorg Med Chem ; 23(3): 602-11, 2015 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-25537531

RESUMO

Naturally occurring condensed quinolines have anticancer properties. In efforts to find active analogues, we designed and synthesized eight polycyclic heterocycles with a pyrimido[1″,2″:1,5]pyrazolo[3,4-b]quinoline framework (IND series). The compounds were evaluated for activity against colon (HCT-116 and S1-MI-80), prostate (PC3 and DU-145), breast (MCF-7 and MDAMB-231), ovarian (ov2008 and A2780), and hepatocellular (HepG2) cancer cells and against non-cancerous Madin Darby canine kidney (MDCK), mouse embryonic fibroblast (NIH/3T3), and human embryonic kidney cells (HEK293). IND-2, a 4-chloro-2-methyl pyrimido[1″,2″:1,5]pyrazolo[3,4-b]quinoline, exhibited more than ten-fold selectivity and potent cytotoxic activity against colon cancer cells relative to the other cancer and non-cancer cells. With five additional colon cancer cell lines (HT-29, HCT-15, LS-180, LS-174, and LoVo), IND-2 had similar cytotoxicity and selectivity, and sub-micromolar concentrations caused changes in the morphology of HCT-116 and HCT-15 cells. IND-2 did not activate the transactivating function of the pregnane X receptor (PXR), indicating that it does not induce PXR-regulated ABCB1 or ABCG2 transporters. Indeed, IND-2 was not a substrate of ABCB1 or ABCG2, and it induced cytotoxicity in HEK293 cells overexpressing ABCB1 or ABCG2 to the same extent as in normal HEK293 cells. IND-2 was cytotoxic to resistant colon carcinoma S1-MI-80 cells, approximately three- and five-fold more than SN-38 and topotecan, respectively. In HCT-116 colon cancer cells, IND-2 produced concentration-dependent changes in mitochondrial membrane potential, leading to apoptosis, and sub-micromolar concentrations caused chromosomal DNA fragmentation. These findings suggest that, by increasing apoptosis, IND-2 has potential therapeutic efficacy for colorectal cancer.


Assuntos
Apoptose/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Quinolinas/farmacologia , Animais , Antineoplásicos/farmacologia , Cães , Resistência a Múltiplos Medicamentos , Resistencia a Medicamentos Antineoplásicos , Ensaios de Seleção de Medicamentos Antitumorais , Células HCT116 , Células HEK293 , Células Hep G2 , Humanos , Células MCF-7 , Células Madin Darby de Rim Canino , Camundongos , Pirimidinas/farmacologia
16.
In Vivo ; 28(1): 67-73, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24425838

RESUMO

BACKGROUND: Emerging interest on three-dimensional (3D) cell culture models to replace two-dimensional cultures of cancer cells and their xenografts in immunocompromised animal hosts prompted us to investigate the use of new biodegradable gels to recapitulate the physiological conditions of the microenvironment of multiple myeloma (MM) cells. MATERIALS AND METHODS: In the present study, for the first time, we used a new 3D model of hyaluronic acid (HA)-based hydrogels with difference in their matrix composition and stiffness. RESULTS: We demonstrated that hyaluronic acid (HA)-based hydrogels perfectly accommodate MM cells; confirmed by cell survival, migration, colony forming units and expression of cell adhesion proteins of the Wnt signaling pathways over a period of time. CONCLUSION: This study provides the first 3D microenvironment data that HA-based hydrogels could provide with a suitable 3D substratum for MM cells to comprehensively analyze phenotypic changes and the influence of bone marrow stromal stem cells on Wnt/ß catenin signaling in response to targeted drug treatments.


Assuntos
Técnicas de Cultura de Células , Células Clonais , Mieloma Múltiplo/genética , Via de Sinalização Wnt/genética , Humanos , Ácido Hialurônico/metabolismo , Hidrogel de Polietilenoglicol-Dimetacrilato , Mieloma Múltiplo/patologia
17.
Anticancer Res ; 32(11): 4697-707, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23155232

RESUMO

BACKGROUND: Development and progression of multiple myeloma is dependent on the bone marrow (BM) microenvironment, and within the BM, a number of factors are secreted, including the Wnt ligands. Bone marrow stromal cells (BMSC) secrete Wnt ligands that activate Wnt signaling in multiple myeloma. The canonical Wnt pathway which is mediated through the transcriptional effector ß-catenin (ß-cat) is commonly de-regulated in many cancers. Cells with active ß-cat-regulated transcription (CRT) are protected against apoptosis; conversely, inhibition of CRT may prevent cell proliferation. MATERIALS AND METHODS: In this study, we tested the efficacy of recently described inhibitors of CRT (iCRTs; oxazole and thiazole) for their selective antagonistic effect on Wnt-ß-cat response in MM cells MM.1, U266, BMSC and primary BMMC obtained from patient samples (n=16). RESULTS: We demonstrated that iCRTs we used, block Wnt/ß-cat reporter activity, down regulate ß-cat expression and inhibit cell proliferation in a dose-dependent manner with an optimal dose closer to 15 µM. Our data further indicate that iCRTs do not influence the expression of the upstream components of the Wnt pathway DKK1 at the optimal dose, suggesting that iCRTs may specifically target ß-cat in MM cells. Additionally, iCRT-treatment of MM cells, co-cultured with BMSC, showed an inhibitory effect on VEGF and cell migration. CONCLUSION: This study provides the first in vitro data evaluation of newly-described iCRTs as potential Wnt-ß-cat/VEGF pathway antagonists in multiple myeloma.


Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Mieloma Múltiplo/metabolismo , Transdução de Sinais/efeitos dos fármacos , Proteínas Wnt/antagonistas & inibidores , beta Catenina/antagonistas & inibidores , Western Blotting , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Humanos , Mieloma Múltiplo/tratamento farmacológico , Mieloma Múltiplo/patologia , Oxazóis/farmacologia , Reação em Cadeia da Polimerase em Tempo Real , Tiazóis/farmacologia , Transfecção , Fator A de Crescimento do Endotélio Vascular/biossíntese
18.
Nano Lett ; 12(11): 5616-21, 2012 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-23030825

RESUMO

Here, we design and develop high-power electric double-layer capacitors (EDLCs) using carbon-based three dimensional (3-D) hybrid nanostructured electrodes. 3-D hybrid nanostructured electrodes consisting of vertically aligned carbon nanotubes (CNTs) on highly porous carbon nanocups (CNCs) were synthesized by a combination of anodization and chemical vapor deposition techniques. A 3-D electrode-based supercapacitor showed enhanced areal capacitance by accommodating more charges in a given footprint area than that of a conventional CNC-based device.

19.
Front Plant Sci ; 3: 171, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22993514

RESUMO

We have engineered the tropical root crop cassava (Manihot esculenta) to express the Chlamydomonas reinhardtii iron assimilatory gene, FEA1, in its storage roots with the objective of enhancing the root nutritional qualities. Iron levels in mature cassava storage roots were increased from 10 to 36 ppm in the highest iron accumulating transgenic lines. These iron levels are sufficient to meet the minimum daily requirement for iron in a 500 g meal. Significantly, the expression of the FEA1 gene in storage roots did not alter iron levels in leaves. Transgenic plants also had normal levels of zinc in leaves and roots consistent with the specific uptake of ferrous iron mediated by the FEA1 protein. Relative to wild-type plants, fibrous roots of FEA1 expressing plants had reduced Fe (III) chelate reductase activity consistent with the more efficient uptake of iron in the transgenic plants. We also show that multiple cassava genes involved in iron homeostasis have altered tissue-specific patterns of expression in leaves, stems, and roots of transgenic plants consistent with increased iron sink strength in transgenic roots. These results are discussed in terms of strategies for the iron biofortification of plants.

20.
Plant Sci ; 195: 151-6, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22921009

RESUMO

In diploid segregation, each alternative allele has a 50% chance of being passed on to the offspring. Mutations in genes involved in the process of meiotic division or early stages of reproductive cell development can affect allele frequency in the gametes. In addition, competition among gametes and differential survival rates of gametes can lead to segregation distortion. In a recent transformation study, a male-sterile, female-sterile (MSFS) mutant was identified in the soybean cultivar, Williams. The mutant in heterozygous condition segregated 3 fertile:1 sterile in the progeny confirming monogenic inheritance. To map the lesion, we generated an F(2) mapping population by crossing the mutant (in heterozygous condition) with Minsoy (PI 27890). The F(2) progeny showed strong segregation distortion against the MSFS phenotype. The objectives of our study were to molecularly map the gene responsible for sterility in the soybean genome, to determine if the MSFS gene is a result of T-DNA insertion during Agrobacterium-mediated transformation, and to map the region that showed distorted segregation. The fertility/sterility locus was mapped to molecular linkage group (MLG) D1a (chromosome Gm01) using bulked segregant analysis. The closest marker, Satt531, mapped 9.4cM from the gene. Cloning of insertion sites for T-DNA in the mutant plants revealed that there are two copies of T-DNA in the genome. Physical locations of these insertion sites do not correlate with the map location of the MSFS gene, suggesting that MSFS mutation may not be associated with T-DNA insertions. Segregation distortion was most extreme at or around the st_A06-2/6 locus suggesting that sterility and segregation distortion are tightly linked attributes. Our results cue that the distorted segregation may be due to a gamete elimination system.


Assuntos
Ligação Genética , Loci Gênicos , Células Germinativas Vegetais/fisiologia , Glycine max/genética , Meiose/genética , Mutação , Infertilidade das Plantas/genética , Agrobacterium , Alelos , Mapeamento Cromossômico , Cromossomos de Plantas , Cruzamentos Genéticos , DNA Bacteriano , Genoma de Planta , Heterozigoto , Padrões de Herança , Mutagênese Insercional , Fenótipo , Infertilidade das Plantas/fisiologia , Reprodução , Glycine max/fisiologia , Transformação Genética
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