WorldWideScience

Sample records for bone protein extract

  1. Calcium Sulfate with Stearic Acid as an Encouraging Carrier for Reindeer Bone Protein Extract

    Directory of Open Access Journals (Sweden)

    Pekka Jalovaara

    2011-07-01

    Full Text Available Various bone proteins and growth factors in specific concentrations are required for bone formation. If the body cannot produce sufficient quantities of these factors, bone trauma can be healed with an implant that includes the required factors in a carrier. This study was designed to evaluate various calcium salt candidates that can be used as carrier with reindeer bone protein extract to induce ectopic bone formation in the muscle pouch model of mouse. The bone protein extract was either impregnated into the disc form of carrier or mixed with carrier powder before implantation. The radiographic analysis indicated increased bone formation in all of the active groups containing the bone protein extract compared to the controls within 21 days follow-up. The highest bone formation was seen in the group with calcium sulfate with stearic acid where new bone and calcified cartilage were clearly visible. The greatest bone formation occurred in the groups that had bone protein extract readily available. This indicates that the bone forming factors in sufficient concentrations are required at the early stage of bone formation. The calcium sulfate with stearic acid was the most suitable and effective carrier for reindeer bone protein extract.

  2. Osteoinductivity assay of the variability of repeated extractions of bone morphogenetic proteins from bovine bone at different times

    Institute of Scientific and Technical Information of China (English)

    HU Zhen-ming 胡侦明; Sean AF Peel; Cameron ML Clokie

    2004-01-01

    Objective:To observe the activity of repeated extracts of bone matrix and the production of purified bone morphogenetic proteins (BMPs).Methods: BMPs were extracted 1- 4 times from fresh bovine cortical bone by the modified Urist's method, with each collected precipitate separated and lyophilized as partially purified BMPs. Another fresh bovine bone was extracted three times and the precipitates were mixed and lyophilized. Meanwhile, the alkaline phosphatase (ALP)activity was measured by an in vitro assay employing cultured C2C12 mouse myoblast cells through the osteoinductivity of bovine BMPs extracted four times at days 1, 4, 7, and 14, and the correlation between BMPs quantities and costing during extraction processes was analyzed.Results:The purified and the cost showed a positive correlation(r=0.969).To separate and lyophilize each collected precipitate as partially purified BMPs raised the cost,and mixed precipitates also cost much.ALPactivities of 1st and mixed extractions of BMPs were shown to be highly osteoinductive and keep a significantly high level(P<0.05-0.01)4 days after culturing compared with the 2nd,3rd and 4th extractions,especially the control group.However,the more times the extraction ws done,the less activity of BMPs was shown and more costing was.The x-ray and histological analysis also showed that the 1st extraction of BMPs induced more ossicles and new bone formation.Conclusions:The results indicated that BMPs enhanced the abilities of osteoinductiviyt in C2C12 culture in vitro.The first extraction of BMPsfrom bone is fitfull,4th extractions are unnecessary for they cost more and waste more time,say nothing of mixed extractions.

  3. EFFECT OF TEMPERATURE AND PARTICLE SIZE ON THE ALKALINE EXTRACTION OF PROTEIN FROM CHICKEN BONE WASTE

    Directory of Open Access Journals (Sweden)

    Andri Cahyo Kumoro

    2012-02-01

    Full Text Available Chicken bone is a waste of chicken meat processing industry and restaurants that has not been used widely, even though it contains valuable organic compounds that are functionals, such as collagenous and non collagenous protein. This research was conducted to investigate the effect of temperature and particle size on the protein extraction from chicken bones using dilute sodium hydroxide solution. Controlled parameters in this study were the solvent in the form of sodium hydroxide solution, extraction time for 1 hour, pH 10.5, the ratio of chicken bone powder: solvent (1:4 w/v, and stirring speed 200 rpm. While the operating variables included the extraction temperature of 30oC, 55oC, and 80oC, and particle size of 150 and 250 μm. Experiments were carried out by heating of 300 mL of sodium hydroxide solution with pH 10.5 in a three-necked flask equipped with Leibig condenser, thermometer, mechanical agitator and sampling device to reach the desired temperature (30oC, 55oC, and 80oC. Then, a total of 75 g of chicken bone powders with desired particle size (150μm and 250μm was introduced into the sodium hydroxide solution and the stirrer was operated at speed of 200rpm. At every 10 minutes interval, as much as 10 mL samples were withdrawn for total protein analysis using Lowry-Folin method. The experiment was terminated after 1 hour. The results show that both increase in temperature and particle size caused an increase in the amount of extracted protein. Highest concentration of protein extracted was achieved at 630.99 mg/L, when the extraction was carried out using 250mm bone particles and temperature 80oC.

  4. On one method of fat and protein extraction from bone mass

    International Nuclear Information System (INIS)

    This article describes the actual technological task of the food industry. The problem of the extraction of fat and protein from the bone mass can be solved by different methods. The work offers one of the more effective modes. Results are presented as diagrams. (author)

  5. Effect of sterilization and delivery systems on the osteoinductivity of reindeer bone morphogenetic protein extract

    OpenAIRE

    Pekkarinen, T. (Tarmo)

    2005-01-01

    Abstract Bone morphogenetic proteins (BMPs) constitute a large family of osteoinductive proteins. Different BMPs are widely used in animal experiments and increasingly in the field of bone surgery. However, the sterilization of BMPs and the choice of a suitable mode of delivery, which binds and slowly releases BMP molecules, are still under intensive investigation. The aims of this study were to evaluate the effects of ethylene oxide and gamma sterilizations and different delivery syst...

  6. Determination of protein content in chicken bone extract%鸡骨素中蛋白质含量的测定

    Institute of Scientific and Technical Information of China (English)

    魏永义; 胡凯

    2015-01-01

    对鸡骨素中的蛋白质含量进行测定,结果表明所测样品中蛋白质含量为24.8g/100g,该方法简单、准确度高,适用于鸡骨素中的蛋白质含量的测定。%The protein content of chicken bone extracts was determined. The result showed that the protein content of the sample was 24. 8g/100g. This method was simple, high accuracy and suitable for the determination of protein content in chicken bone extract.

  7. Recombinant human bone morphogenetic protein induces bone formation

    International Nuclear Information System (INIS)

    The authors have purified and characterized active recombinant human bone morphogenetic protein (BMP) 2A. Implantation of the recombinant protein in rats showed that a single BMP can induce bone formation in vivo. A dose-response and time-course study using the rat ectopic bone formation assay revealed that implantation of 0.5-115 μg of partially purified recombinant human BMP-2A resulted in cartilage by day 7 and bone formation by day 14. The time at which bone formation occurred was dependent on the amount of BMP-2A implanted; at high doses bone formation could be observed at 5 days. The cartilage- and bone-inductive activity of the recombinant BMP-2A is histologically indistinguishable from that of bone extracts. Thus, recombinant BMP-2A has therapeutic potential to promote de novo bone formation in humans

  8. Bone formation following implantation of bone biomaterials into extraction sites

    OpenAIRE

    Molly, Liene; Vandromme, Heleen; Quirynen, Marc; Schepers, Evert; Adams, Jessica L; van Steenberghe, Daniel

    2008-01-01

    Background: Adequate bone volume is imperative for the osseointegration of endosseous implants, but post-extraction resorption and remodeling may challenge implant placement. The use of bone biomaterials has been advocated to fill extraction sites and to enhance primary implant stability during osseointegration. The objective of the case series was to evaluate bone formation histologically and biomechanically in extraction sites following implantation of three commercially available bone biom...

  9. Peptide-induced de novo bone formation after tooth extraction prevents alveolar bone loss in a murine tooth extraction model.

    Science.gov (United States)

    Arai, Yuki; Aoki, Kazuhiro; Shimizu, Yasuhiro; Tabata, Yasuhiko; Ono, Takashi; Murali, Ramachandran; Mise-Omata, Setsuko; Wakabayashi, Noriyuki

    2016-07-01

    Tooth extraction causes bone resorption of the alveolar bone volume. Although recombinant human bone morphogenetic protein 2 (rhBMP-2) markedly promotes de novo bone formation after tooth extraction, the application of high-dose rhBMP-2 may induce side effects, such as swelling, seroma, and an increased cancer risk. Therefore, reduction of the necessary dose of rhBMP-2 which can still obtain sufficient bone mass is necessary by developing a new osteogenic reagent. Recently, we showed that the systemic administration of OP3-4 peptide, which was originally designed as a bone resorption inhibitor, had osteogenic ability both in vitro and in vivo. This study evaluated the ability of the local application of OP3-4 peptide to promote bone formation in a murine tooth extraction model with a very low-dose of BMP. The mandibular incisor was extracted from 10-week-old C57BL6/J male mice and a gelatin hydrogel containing rhBMP-2 with or without OP3-4 peptide (BMP/OP3-4) was applied to the socket of the incisor. Bone formation inside the socket was examined radiologically and histologically at 21 days after the extraction. The BMP/OP3-4-group showed significant bone formation inside the mandibular extraction socket compared to the gelatin-hydrogel-carrier-control group or rhBMP-2-applied group. The BMP/OP3-4-applied mice showed a lower reduction of alveolar bone and fewer osteoclast numbers, suggesting that the newly formed bone inside the socket may prevent resorption of the cortical bone around the extraction socket. Our data revealed that OP3-4 peptide promotes BMP-mediated bone formation inside the extraction socket of mandibular bone, resulting in preservation from the loss of alveolar bone. PMID:27118173

  10. Bone morphogenetic proteins: Periodontal regeneration

    Directory of Open Access Journals (Sweden)

    Subramaniam M Rao

    2013-01-01

    Full Text Available Periodontitis is an infectious inflammatory disease that results in attachment loss and bone loss. Regeneration of the periodontal tissues entails de novo formation of cementum, periodontal ligament, and alveolar bone. Several different approaches are currently being explored to achieve complete, reliable, and reproducible regeneration of periodontal tissues. The therapeutic management of new bone formation is one of the key issues in successful periodontal regeneration. Bone morphogenetic proteins form a unique group of proteins within the transforming growth factor superfamily of genes and have a vital role in the regulation in the bone induction and maintenance. The activity of bone morphogenetic proteins was first identified in the 1960s, but the proteins responsible for bone induction were unknown until the purification and cloning of human bone morphogenetic proteins in the 1980s, because of their osteoinductive potential. Bone morphogenetic proteins have gained a lot of interest as therapeutic agents for treating periodontal defects. A systematic search for data related to the use of bone morphogenetic proteins for the regeneration of periodontal defects was performed to recognize studies on animals and human (PUBMED, MEDLINE, COCHRANE, and Google search. All the studies included showed noticeable regeneration of periodontal tissues with the use of BMP.

  11. Time-sequential changes of differentially expressed miRNAs during the process of anterior lumbar interbody fusion using equine bone protein extract, rhBMP-2 and autograft

    Science.gov (United States)

    Chen, Da-Fu; Zhou, Zhi-Yu; Dai, Xue-Jun; Gao, Man-Man; Huang, Bao-Ding; Liang, Tang-Zhao; Shi, Rui; Zou, Li-Jin; Li, Hai-Sheng; Bünger, Cody; Tian, Wei; Zou, Xue-Nong

    2014-03-01

    The precise mechanism of bone regeneration in different bone graft substitutes has been well studied in recent researches. However, miRNAs regulation of the bone formation has been always mysterious. We developed the anterior lumbar interbody fusion (ALIF) model in pigs using equine bone protein extract (BPE), recombinant human bone morphogenetic protein-2 (rhBMP-2) on an absorbable collagen sponge (ACS), and autograft as bone graft substitute, respectively. The miRNA and gene expression profiles of different bone graft materials were examined using microarray technology and data analysis, including self-organizing maps, KEGG pathway and Biological process GO analyses. We then jointly analyzed miRNA and mRNA profiles of the bone fusion tissue at different time points respectively. Results showed that miRNAs, including let-7, miR-129, miR-21, miR-133, miR-140, miR-146, miR-184, and miR-224, were involved in the regulation of the immune and inflammation response, which provided suitable inflammatory microenvironment for bone formation. At late stage, several miRNAs directly regulate SMAD4, Estrogen receptor 1 and 5-hydroxytryptamine (serotonin) receptor 2C for bone formation. It can be concluded that miRNAs play important roles in balancing the inflammation and bone formation.

  12. The classic: Bone morphogenetic protein.

    Science.gov (United States)

    Urist, Marshall R; Strates, Basil S

    2009-12-01

    This Classic Article is a reprint of the original work by Marshall R. Urist and Basil S. Strates, Bone Morphogenetic Protein. An accompanying biographical sketch of Marshall R. Urist, MD is available at DOI 10.1007/s11999-009-1067-4; a second Classic Article is available at DOI 10.1007/s11999-009-1069-2; and a third Classic Article is available at DOI 10.1007/s11999-009-1070-9. The Classic Article is copyright 1971 by Sage Publications Inc. Journals and is reprinted with permission from Urist MR, Strates BS. Bone morphogenetic protein. J Dent Res. 1971;50:1392-1406. PMID:19727989

  13. A novel method for beef bone protein extraction by lipase-pretreatment and its application in the Maillard reaction.

    Science.gov (United States)

    Song, Shiqing; Li, Sisi; Fan, Li; Hayat, Khizar; Xiao, Zuobing; Chen, Lihua; Tang, Qi

    2016-10-01

    Five beef bone hydrolysates were obtained by different enzyme treatment schemes, including papain (M), combination of porcine pancreatic lipase and papain (Z+M, combination of lipase and papain (Y+M), Protamex (F), combination of porcine pancreatic lipase and Protamex (Z+F). The degree of hydrolysis (DH), free amino acids and molecular weight distribution of these hydrolysates were evaluated. To further explore the differences between these five hydrolysates, Maillard reaction products (MRPs) were prepared using a xylose/cysteine/hydrolysate model. It was found that the DH, content of low molecular weight peptides and amino acids of hydrolysates increased significantly after lipase pre-treatment. GC-MS showed that the total content of furans, pyrroles and thioethers in MRPs Y+M increased by 78.0% compared with MRPs M, while in MRPs Z+F, pyrazines increased by 44.1% compared with MRPs F. Examining the sensory characteristics of the MRPs, the MRP from the hydrolysate of Y+M had the best mouthful, umami and meaty characteristics. The correlation analysis further confirmed that an appropriate lipase pre-treatment could improve the flavour of MRPs. PMID:27132826

  14. Bone Regeneration Using Bone Morphogenetic Proteins and Various Biomaterial Carriers

    Directory of Open Access Journals (Sweden)

    Zeeshan Sheikh

    2015-04-01

    Full Text Available Trauma and disease frequently result in fractures or critical sized bone defects and their management at times necessitates bone grafting. The process of bone healing or regeneration involves intricate network of molecules including bone morphogenetic proteins (BMPs. BMPs belong to a larger superfamily of proteins and are very promising and intensively studied for in the enhancement of bone healing. More than 20 types of BMPs have been identified but only a subset of BMPs can induce de novo bone formation. Many research groups have shown that BMPs can induce differentiation of mesenchymal stem cells and stem cells into osteogenic cells which are capable of producing bone. This review introduces BMPs and discusses current advances in preclinical and clinical application of utilizing various biomaterial carriers for local delivery of BMPs to enhance bone regeneration.

  15. The Sclerostin-Bone Protein Interactome

    OpenAIRE

    Devarajan-Ketha, Hemamalini; Craig, Theodore A.; Madden, Benjamin J.; Bergen, H. Robert; Kumar, Rajiv

    2011-01-01

    The secreted glycoprotein, sclerostin alters bone formation. To gain insights into the mechanism of action of sclerostin, we examined the interactions of sclerostin with bone proteins using a sclerostin affinity capture technique. Proteins from decalcified rat bone were captured on sclerostin-maltose binding protein (MBP) amylose column, or on a MBP amylose column. The columns were extensively washed with low ionic strength buffer, and bound proteins were eluted with buffer containing 1M sodi...

  16. Positive modulator of bone morphogenic protein-2

    Science.gov (United States)

    Zamora, Paul O.; Pena, Louis A.; Lin, Xinhua; Takahashi, Kazuyuki

    2009-01-27

    Compounds of the present invention of formula I and formula II are disclosed in the specification and wherein the compounds are modulators of Bone Morphogenic Protein activity. Compounds are synthetic peptides having a non-growth factor heparin binding region, a linker, and sequences that bind specifically to a receptor for Bone Morphogenic Protein. Uses of compounds of the present invention in the treatment of bone lesions, degenerative joint disease and to enhance bone formation are disclosed.

  17. STUDIES ON THE TECHNOLOGCAL PARAMETERS OF ALKALI EXTRACTION PROCESS FOR RECOVERING PROTEIN FROM SWINE BONES%碱提法回收猪骨蛋白质的探讨

    Institute of Scientific and Technical Information of China (English)

    庞杰; 林娇芬; 陆则坚

    2001-01-01

    对碱提法回收猪骨蛋白质工艺参数的研究表明,4种工艺参数对蛋白质得率均有不同程度的影响。其顺序为浸提液pH>浸提时间>沉降pH>浸提温度。碱提法回收猪骨蛋白质的最高得率为25.7%,最佳工艺参数为:浸提时间105 min,浸提液pH12.0,浸提温度30 ℃和沉降pH5.0。%In this study,the recovery rate of protein from swine bones was found to be influenced by all the four parameters investigated.The significance of their influence,however,was different,being in the order of solvent pH>extraction time> precipitation pH>extraction temperature.The highest recovery rate of protein obtained in this experiment was 25.7 % of the original protein present in te raw bone.The following is recommended as the optimum combination of the four parameters:extraction time 105 min,solvent pH12.0,extraction temperature 30 ℃ and precipitation pH5.0.

  18. Comparative experiment of four different materials as carriers of Bone morphogenetic protein to repair long bone defect

    Institute of Scientific and Technical Information of China (English)

    WEI Kuan-hai; PEI Guo-xian; YANG Run-gong

    2001-01-01

    @@ OBJECTIVE To investigate the effects of four different materials as carriers of bone morphogenetic protein (BMP) to repair long bone defect. METHODS 12 mm radius bone defects were made. They were divided into 4 groups in random and repaired respectively with the vascular muscle flap combined with FS/BMP (group A), vascular muscle flap/BMP (group B), bloodless muscle flap/BMP (group C) and autolyzed antigen-extracted allogeneic bone (AAA)/BMP (group D).Their abilities of bone forming to repair bone defects were observed.

  19. 高压脉冲电场协同酶法提取牛骨蛋白的研究%Research on extracting protein from cattle bone by enzymolysis coupled with PEF

    Institute of Scientific and Technical Information of China (English)

    韦汉昌; 韦善清; 何建华

    2011-01-01

    The technology of extracting protein from cattle bone by flavor protease coupled with pulsed electric field(PEF)was researched.For extracting protein from cattle bone powder pre-treated by PEF,the results of experiments showed that it not only could make the enzymolysis time short but also increase the dissolution ratio of protein.Under the conditions of electric field intensity 30 kV·cm-1,pulse number 25 times,material to liquid ratio 1∶ 8(g/mL),enzymes usage 4000U/g,enzymolysis temperature 50℃,pH=6.5 and enzymolysis time 30min,the dissolution ratio of bone protein could reach 65.2%.%以新鲜牛骨为原料,研究高压脉冲电场协同风味酶提取牛骨蛋白。实验结果表明,牛骨粉经高压脉冲电场处理再进行酶解,不仅能够提高骨蛋白溶出率,而且能缩短酶解时间。在电场强度30kV.cm-1、脉冲数25、料液比1∶8、酶用量4000U/g(骨粉质量)、酶解温度50℃、pH6.5等条件下,酶解30min,牛骨蛋白溶出率可达65.2%。

  20. High-Frequency Acceleration: Therapeutic Tool to Preserve Bone following Tooth Extractions.

    Science.gov (United States)

    Alikhani, M; Lopez, J A; Alabdullah, H; Vongthongleur, T; Sangsuwon, C; Alikhani, M; Alansari, S; Oliveira, S M; Nervina, J M; Teixeira, C C

    2016-03-01

    A common problem in clinical dentistry is the significant and rapid bone loss that occurs after tooth extraction. Currently there is no solution for the long-term preservation of alveolar bone. Previously, we showed that high-frequency acceleration (HFA) has an osteogenic effect on healthy alveolar bone. However, it is not known if HFA can preserve alveolar bone after extraction without negatively affecting wound healing. The purpose of this study was to evaluate the effect of HFA on alveolar bone loss and the rate of bone formation after tooth extraction. Eighty-five adult Sprague-Dawley rats were divided into 3 groups: control, static (static load), and HFA. In all groups, the maxillary right third molar was extracted. The HFA group received HFA for 5 min/d, applied through the second molar. The static group received the same magnitude of static load. The control group did not receive any stimulation. Some animals received fluorescent dyes at 26 and 54 d. Samples were collected on days 0, 7, 14, 28, and 56 for fluorescence microscopy, micro-computed tomography, histology, RNA, and protein analyses. We found that HFA increased bone volume in the extraction site and surrounding alveolar bone by 44% when compared with static, while fully preserving alveolar bone height and width long-term. These effects were accompanied by increased expression of osteogenic markers and intramembranous bone formation and by decreased expression of osteoclastic markers and bone resorption activity, as well as decreased expression of many inflammatory markers. HFA is a noninvasive safe treatment that can be used to prevent alveolar bone loss and/or accelerate bone healing after tooth extraction. PMID:26672126

  1. Enhanced release of bone morphogenetic proteins from demineralized bone matrix by gamma irradiation

    International Nuclear Information System (INIS)

    Gamma irradiation is a useful method for sterilizing demineralized bone matrix (DBM), but its effect on the osteoinductivity of DBM is still controversial. In this study, the osteoinductive activity of gamma-irradiated DBM was examined using a mouse myoblastic cell line (C2C12). DBM was extracted from adult bovine bone and was irradiated at a dose of 25 kGy using a 60cobalt gamma-irradiator. Cell proliferation with DBM was not affected by gamma-irradiation, but alkaline phosphatase and osteocalcin productions were significantly increased in C2C12 cell groups treated with gamma-irradiated DBM. It was reasoned that bone morphogenetic proteins were more efficiently released from gamma-irradiated DBM than from the non-irradiated control. This result suggests the effectiveness of radiation sterilization of bone implants - Highlights: • Demineralized bone matrix (DBM) was gamma-irradiated for sterilization. • Irradiated DBM had higher alkaline phosphatase and osteocalcin production. • It was reasoned the more released bone morphogenetic proteins by irradiation. • This result supports the application of radiation sterilization for bone implants

  2. Study on extract and enzymolysis technology of protein from discarded fish bone%废弃鱼骨中蛋白质的提取分离及酶解工艺研究

    Institute of Scientific and Technical Information of China (English)

    张振兴; 刘永峰; 刘毅; 裴栋; 魏鉴腾; 邸多隆

    2014-01-01

    Different extraction methods and different protease digestion of discarded fish bone proteins were studied using extraction rate of proteins and molecular weight of peptides as evaluation indexes. The results showed that the method of high-pressure cooking had a significantly higher protein content of 86.15% and protein extraction rate of 16.76% than the methods of constant temperature water bath and hot reflux extraction. The neutral protease had a higher activity than other three kinds of proteases. The protein content could reach 88.46% and the peptide ratio could reach 95% in the protein hydrolysates using neutral protease. The enzyme dosage could be reduced when the protein was extracted with high-pressure cooking method and the polypeptide was prepared with neutral protease. This method could provide reference and new idea for polypeptide industrial production.%以蛋白提取率和多肽分子质量分布为评价指标,采用不同提取方法和不同蛋白酶对废弃鱼骨蛋白质的提取和酶解工艺进行了研究。结果表明,高压蒸煮法提取的鱼骨蛋白质中蛋白含量为86.15%,蛋白提取率为16.76%,明显高于恒温水浴和热回流提取法。中性蛋白酶酶解的多肽中蛋白含量为88.46%,多肽比例高达95%以上,明显高于其他三种酶。先利用高压提取法提取蛋白质,再利用中性蛋白酶酶解蛋白质制备多肽,可降低酶用量,该方法为多肽工业化生产提供参考及新思路。

  3. Preradiation dental extractions and the incidence of bone necrosis

    International Nuclear Information System (INIS)

    Studies were done with 120 patients submitting to preradiation dental extraction within the radiation treatment volume. Bone necroses developed at the extraction sites in 17 patients (14.1%). The risk of bone necrosis in these patients is primarily dependent upon the size of the radiation treatment volume, radiation dose to mandibular bone, and healing time for the extraction wounds. Of the 13 mandibular bone necroses occurring at preradiation extraction sites, only two have not responded favorably to conservative management. Our data indicates that a policy of selected tooth removal, before radiation treatment, will minimize the risk of osteoradionecrosis. Mandibular molars with advanced chronic periodontal bone loss, residing within the proposed radiation field should be considered for removal before commencement of radiation treatment

  4. Bone Morphogenic Proteins: Applications in Spinal Surgery

    OpenAIRE

    Jeong, Gerard K.; Sandhu, Harvinder S.; Farmer, James

    2005-01-01

    The prospect of predictable and reliable oseteogenesis without the need for secondary bone grafting to treat a wide spectrum of spinal disorders is tremendously appealing. Recombinant human bone morphogenic proteins (rhBMP) have been the subject of extensive basic science, animal, and clinical research as a potential therapeutic modality to promote bony fusion. Animal studies and prospective, randomized clinical trials have demonstrated the efficacy of rhBMPs as an adjunct or substitute to au...

  5. Extraction of high value added gelatin biopolymer from black tilapia (Oreochromis mossambicus) head bones

    Energy Technology Data Exchange (ETDEWEB)

    Sockalingam, K., E-mail: gd130106@siswa.uthm.edu.my; Abdullah, H. Z., E-mail: hasan@uthm.edu.my [Faculty of Mechanical and Manufacturing Engineering, Universiti Tun Hussein Onn Malaysia, 86400 Parit Raja, Batu Pahat, Johor (Malaysia)

    2015-07-22

    Black tilapia (Oreochromis mossambicus) fish head bones were evaluated for its possibilities in extracting gelatin. Head bones were subjected to pre-treatment with 3% of hydrochloric acid (HCl) for demineralization before undergoes thermal extraction process. The raw head bones were characterized via Scanning Electron Microscopy (SEM) in order to investigate the external and internal surface morphology. SEM images also reveal the presence of collagen fiber with 1 µm diameter in the head bone. The black tilapia fish head bones yields 5.75 % of gelatin in wet weight basis, indicating the possibility of this fish species as sources of gelatin. Further characterizations were done on both raw head bones and extracted gelatin through Fourier Transform Infrared Spectroscopy (FTIR) and proximate analysis. The head bones gelatin shows high protein (10.55%) and ash (3.11 %) content with low moisture. This further proves the effectiveness of demineralization and extraction method used. The black tilapia fish head bones are found to be a prospective source of gelatin with good chemical and functional properties.

  6. Extraction of high value added gelatin biopolymer from black tilapia (Oreochromis mossambicus) head bones

    International Nuclear Information System (INIS)

    Black tilapia (Oreochromis mossambicus) fish head bones were evaluated for its possibilities in extracting gelatin. Head bones were subjected to pre-treatment with 3% of hydrochloric acid (HCl) for demineralization before undergoes thermal extraction process. The raw head bones were characterized via Scanning Electron Microscopy (SEM) in order to investigate the external and internal surface morphology. SEM images also reveal the presence of collagen fiber with 1 µm diameter in the head bone. The black tilapia fish head bones yields 5.75 % of gelatin in wet weight basis, indicating the possibility of this fish species as sources of gelatin. Further characterizations were done on both raw head bones and extracted gelatin through Fourier Transform Infrared Spectroscopy (FTIR) and proximate analysis. The head bones gelatin shows high protein (10.55%) and ash (3.11 %) content with low moisture. This further proves the effectiveness of demineralization and extraction method used. The black tilapia fish head bones are found to be a prospective source of gelatin with good chemical and functional properties

  7. Extraction of high value added gelatin biopolymer from black tilapia (Oreochromis mossambicus) head bones

    Science.gov (United States)

    Sockalingam, K.; Abdullah, H. Z.

    2015-07-01

    Black tilapia (Oreochromis mossambicus) fish head bones were evaluated for its possibilities in extracting gelatin. Head bones were subjected to pre-treatment with 3% of hydrochloric acid (HCl) for demineralization before undergoes thermal extraction process. The raw head bones were characterized via Scanning Electron Microscopy (SEM) in order to investigate the external and internal surface morphology. SEM images also reveal the presence of collagen fiber with 1 µm diameter in the head bone. The black tilapia fish head bones yields 5.75 % of gelatin in wet weight basis, indicating the possibility of this fish species as sources of gelatin. Further characterizations were done on both raw head bones and extracted gelatin through Fourier Transform Infrared Spectroscopy (FTIR) and proximate analysis. The head bones gelatin shows high protein (10.55%) and ash (3.11 %) content with low moisture. This further proves the effectiveness of demineralization and extraction method used. The black tilapia fish head bones are found to be a prospective source of gelatin with good chemical and functional properties.

  8. Effects of Eucommia ulmoides extract on longitudinal bone growth rate in adolescent female rats.

    Science.gov (United States)

    Kim, Ji Young; Lee, Jeong-Il; Song, MiKyung; Lee, Donghun; Song, Jungbin; Kim, Soo Young; Park, Juyeon; Choi, Ho-Young; Kim, Hocheol

    2015-01-01

    Eucommia ulmoides is one of the popular tonic herbs for the treatment of low back pain and bone fracture and is used in Korean medicine to reinforce muscles and bones. This study was performed to investigate the effects of E. ulmoides extract on longitudinal bone growth rate, growth plate height, and the expressions of bone morphogenetic protein 2 (BMP-2) and insulin-like growth factor 1 (IGF-1) in adolescent female rats. In two groups, we administered a twice-daily dosage of E. ulmoides extract (at 30 and 100 mg/kg, respectively) per os over 4 days, and in a control group, we administered vehicle only under the same conditions. Longitudinal bone growth rate in newly synthesized bone was observed using tetracycline labeling. Chondrocyte proliferation in the growth plate was observed using cresyl violet dye. In addition, we analyzed the expressions of BMP-2 and IGF-1 using immunohistochemistry. Eucommia ulmoides extract significantly increased longitudinal bone growth rate and growth plate height in adolescent female rats. In the immunohistochemical study, E. ulmoides markedly increased BMP-2 and IGF-1 expressions in the proliferative and hypertrophic zones. In conclusion, E. ulmoides increased longitudinal bone growth rate by promoting chondrogenesis in the growth plate and the levels of BMP-2 and IGF-1. Eucommia ulmoides could be helpful for increasing bone growth in children who have growth retardation. PMID:25087723

  9. Does crystallinity of extracted bone mineral increase over storage time?

    Directory of Open Access Journals (Sweden)

    William Querido

    2013-01-01

    Full Text Available It was recently shown that the crystallinity of extracted bone mineral samples from the fin bones of zebrafish could increase over storage time. This would have implications in many studies in which the samples need to be stored until analysis. The aim of this study was to further evaluate if the crystallinity of extracted bone mineral increases over storage time. The extracted mineral was a biological bone-like apatite produced in osteoblast cell cultures. The overall characterization of the mineral was done by energy dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and high-resolution transmission electron microscopy. In order to evaluate possible changes in crystallinity over storage time, the same sample was analyzed by X-ray diffraction immediately after mineral extraction and after 18 months of storage. In conclusion, no statistically relevant changes were observed over storage time, although the occurrence of a slight increase in crystallinity could be discussed in the stored mineral sample.

  10. Automatic Extraction of Protein Interaction in Literature

    OpenAIRE

    Liu, Peilei; Wang, Ting

    2014-01-01

    Protein-protein interaction extraction is the key precondition of the construction of protein knowledge network, and it is very important for the research in the biomedicine. This paper extracted directional protein-protein interaction from the biological text, using the SVM-based method. Experiments were evaluated on the LLL05 corpus with good results. The results show that dependency features are import for the protein-protein interaction extraction and features related to the interaction w...

  11. Effect of biomaterial properties on bone healing in a rabbit tooth extraction socket model.

    Science.gov (United States)

    Fisher, John P; Lalani, Zahid; Bossano, Carla M; Brey, Eric M; Demian, Nagi; Johnston, Carol M; Dean, David; Jansen, John A; Wong, Mark E K; Mikos, Antonios G

    2004-03-01

    In this work we sought to understand the effect of biomaterial properties upon healing bone tissue. We hypothesized that a hydrophilic polymer gel implanted into a bone tissue defect would impede the healing process owing to the biomaterial's prevention of protein adsorption and thus cell adhesion. To test this hypothesis, healing bone was investigated within a rabbit incisor extraction socket, a subcritical size bone defect that resists significant soft tissue invasion by virtue of its conformity. After removal of the incisor teeth, one tooth socket was left as an empty control, one was filled with crosslinked polymer networks formed from the hydrophobic polymer poly(propylene fumarate) (PPF), and one was filled with a hydrogel formed from the hydrophilic oligomer oligo(poly(ethylene glycol) fumarate) (OPF). At five different times (4 days as well as 1, 2, 4, and 8 weeks), jaw bone specimens containing the tooth sockets were removed. We analyzed bone healing by histomorphometrical analysis of hematoxylin and eosin stained sections as well as immunohistochemically stained sections. The proposed hypothesis, that a hydrophilic material would hinder bone healing, was supported by the histomorphometrical results. In addition, the immunohistochemical results reflect molecular signaling indicative of the early invasion of platelets, the vascularization of wound-healing tissue, the differentiation of migrating progenitor cells, and the formation and remodeling of bone tissue. Finally, the results emphasize the need to consider biomaterial properties and their differing effects upon endogenous growth factors, and thus bone healing, during the development of tissue engineering devices. PMID:14762922

  12. Bone graft substitutes and bone morphogenetic proteins for osteoporotic fractures: What is the evidence?

    NARCIS (Netherlands)

    E.M.M. van Lieshout (Esther); V. Alt (Volker)

    2016-01-01

    textabstractDespite improvements in implants and surgical techniques, osteoporotic fractures remain challenging to treat. Among other major risk factors, decreased expression of morphogenetic proteins has been identified for impaired fracture healing in osteoporosis. Bone grafts or bone graft substi

  13. Chemical techniques to extract organic fractions from fossil bones for accurate 14C dating

    International Nuclear Information System (INIS)

    We examined different concentrations of HCl, such as 0.4, 0.6, 0.8, 1.0 and 1.2 M, for decalcification of fossil bones and different times of 0.1 M NaOH treatment on collagens to determine the best conditions for purifying collagen through extraction of humic contaminants, and compared the alkali treatment method with the XAD-2 treatment method for several types of fossils. The yield of acid-insoluble bone fractions did not change over the range from 0.4 to 1.0 M HCl and decreased suddenly with 1.2 M HCl on decalcification, and the 14C ages of the extracted gelatins from the five decalcified fractions were unchanged, suggesting that 14C ages as those of the XAD-purified hydrolysates. The NaOH-treatment time should be less than several hours to avoid a loss of collagen. The fossil bones used are relatively well-preserved, but the alkali treatment could bring about a lot of loss of organic bone proteins for poorly-preserved bones. The XAD-2 treatment method is effective for accurate radiocarbon dating of fossil bones, if the XAD-2 resin is completely pre-cleaned

  14. Rhus javanica Gall Extract Inhibits the Differentiation of Bone Marrow-Derived Osteoclasts and Ovariectomy-Induced Bone Loss

    Directory of Open Access Journals (Sweden)

    Tae-Ho Kim

    2016-01-01

    Full Text Available Inhibition of osteoclast differentiation and bone resorption is a therapeutic strategy for the management of postmenopausal bone loss. This study investigated the effects of Rhus javanica (R. javanica extracts on bone marrow cultures to develop agents from natural sources that may prevent osteoclastogenesis. Extracts of R. javanica (eGr cocoons spun by Rhus javanica (Bell. Baker inhibited the osteoclast differentiation and bone resorption. The effects of aqueous extract (aeGr or 100% ethanolic extract (eeGr on ovariectomy- (OVX- induced bone loss were investigated by various biochemical assays. Furthermore, microcomputed tomography (µCT was performed to study bone remodeling. Oral administration of eGr (30 mg or 100 mg/kg/day for 6 weeks augmented the inhibition of femoral bone mineral density (BMD, bone mineral content (BMC, and other factors involved in bone remodeling when compared to OVX controls. Additionally, eGr slightly decreased bone turnover markers that were increased by OVX. Therefore, it may be suggested that the protective effects of eGr could have originated from the suppression of OVX-induced increase in bone turnover. Collectively, the findings of this study indicate that eGr has potential to activate bone remodeling by inhibiting osteoclast differentiation and bone loss.

  15. Multifunctional Bone Morphogenetic Protein System in Endocrinology

    Directory of Open Access Journals (Sweden)

    Otsuka,Fumio

    2013-04-01

    Full Text Available New biological activities of bone morphogenetic proteins (BMPs in the endocrine system have recently been revealed. The BMP system is composed of approximately 30 ligands and preferential combinations of type I and type II receptors. The BMP system not only induces bone formation but also plays unique tissue-specific roles in various organs. For instance, the ovarian BMP system is a physiological inhibitor of luteinization in growing ovarian follicles. In the ovary, the expression of oocyte-derived BMP-15 is critical for female reproduction. In the pituitary, BMP-4 is a key player for initial development of the anterior pituitary, while it is also functionally involved in some differentiated pituitary tumors, including prolactinoma and Cushingʼs disease. In the adrenal glands, BMP-6 and BMP-4 modulate aldosterone and catecholamine production, respectively, which contributes to a functional interaction between the cortex and medulla. In the present review, recent advances in BMP biology in the field of endocrinology are described and the possibility for clinical application of BMP activity is discussed.

  16. Sinonasal Schwannoma with New Bone Formation Expressing Bone Morphogenic Protein

    OpenAIRE

    Satoru Kodama; Tomoyo Okamoto; Masashi Suzuki

    2010-01-01

    Schwannoma is a benign tumor that arises from the sheath of myelinated nerve fibers and may occur in any part of the body. Osteogenesis in schwannoma is extremely rare and, to date, new bone formation in sinonasal schwannoma has not yet been reported. Here, we describe the first reported case of sinonasal schwannoma with new bone formation. The tumor was successfully treated by endoscopic sinus surgery, and the patient showed no evidence of recurrence 24 months postoperatively. Immunohistoche...

  17. Preparation of denatured protein bone sterilized with gamma radiation

    International Nuclear Information System (INIS)

    The bone is one of the tissues more transplanted in the entire world by that the bone necessity for transplant every day becomes bigger. In the Bank of tissues Radio sterilized of the ININ the amnion and the pig skin are routinely processed. The tissue with which will be continued is with bone. Due to that in our country it doesn't have enough bone of human origin for the necessities required in the bone transplant, an option is the bone of bovine. Of this bone one can obtain denatured protein bone, with the same characteristics of the denatured protein human bone, the one which has been proven that it has good acceptance and incorporation in the human body when is transplanted. The method for the obtaining of the denatured protein bone of bovine, with the confirmation of the final product by means of X-ray diffraction is described. The radiosterilization of this bone with gamma rays and the determination of the lead content. (Author)

  18. A strategy to quantitate global phosphorylation of bone matrix proteins.

    Science.gov (United States)

    Sroga, Grażyna E; Vashishth, Deepak

    2016-04-15

    Current studies of protein phosphorylation focus primarily on the importance of specific phosphoproteins and their landscapes of phosphorylation in the regulation of different cellular functions. However, global changes in phosphorylation of extracellular matrix phosphoproteins measured "in bulk" are equally important. For example, correct global phosphorylation of different bone matrix proteins is critical to healthy tissue biomineralization. To study changes of bone matrix global phosphorylation, we developed a strategy that combines a procedure for in vitro phosphorylation/dephosphorylation of fully mineralized bone in addition to quantitation of the global phosphorylation levels of bone matrix proteins. For the first time, we show that it is possible to enzymatically phosphorylate/dephosphorylate fully mineralized bone originating from either cadaveric human donors or laboratory animals (mice). Using our strategy, we detected the difference in the global phosphorylation levels of matrix proteins isolated from wild-type and osteopontin knockout mice. We also observed that the global phosphorylation levels of matrix proteins isolated from human cortical bone were lower than those isolated from trabecular bone. The developed strategy has the potential to open new avenues for studies on the global phosphorylation of bone matrix proteins and their role in biomineralization as well for other tissues/cells and protein-based materials. PMID:26851341

  19. FEATURE EXTRACTION OF BONES AND SKIN BASED ON ULTRASONIC SCANNING

    Institute of Scientific and Technical Information of China (English)

    Zheng Shuxian; Zhao Wanhua; Lu Bingheng; Zhao Zhao

    2005-01-01

    In the prosthetic socket design, aimed at the high cost and radiation deficiency caused by CT scanning which is a routine technique to obtain the cross-sectional image of the residual limb, a new ultrasonic scanning method is developed to acquire the bones and skin contours of the residual limb. Using a pig fore-leg as the scanning object, an overlapping algorithm is designed to reconstruct the 2D cross-sectional image, the contours of the bone and skin are extracted using edge detection algorithm and the 3D model of the pig fore-leg is reconstructed by using reverse engineering technology. The results of checking the accuracy of the image by scanning a cylinder work pieces show that the extracted contours of the cylinder are quite close to the standard circumference. So it is feasible to get the contours of bones and skin by ultrasonic scanning. The ultrasonic scanning system featuring no radiation and low cost is a kind of new means of cross section scanning for medical images.

  20. Effects of ionizing radiation on proteins in demineralized, lyophilized or frozen human bone

    International Nuclear Information System (INIS)

    The aim is the study of the application of ionizing radiation (gamma and electron) as sterilizing agents at doses of 15 kGy, 25 kGy and 50 kGy, the demineralized bone tissue frozen and freeze-dried for use in transplants. Five human femoral diaphysis of different donors demineralized bone tissues were preserved as lyophilized or frozen at - 80 deg C. The samples were divided into non-irradiated groups (control) and irradiated by gamma rays or electron beam. The bone proteins were extracted and used to determine the concentrations of total protein, BMP 2 and 7. It was observed a decrease in total protein concentrations, and BMP 2 and 7. The decrease in total protein concentrations, as compared to respective control groups was significant in the lyophilized and frozen samples irradiated at a dose of 50 kGy gamma radiation and beam electrons with greater than 30% reduction. The significant decrease in the levels of BMP 2 and 7 were also observed in higher doses and especially by electron beam. The reductions in the concentrations of total protein and osteoinductive proteins (BMP 2 and 7), were related to the radiation dose, i.e., increase with higher doses of ionizing radiation type and the type of preservation of the bones. The largest reductions in concentrations were observed in bone irradiated by electron beam and at a dose of 50 kGy. But this type of radiation and this high dose are not usual practice for the sterilization of bone tissue. Keywords: demineralized bone tissue, ionizing radiation, Tissue Bank, BMP 2, BMP 7, bone proteins. (author)

  1. Effects of ionizing radiation on proteins in demineralized, lyophilized or frozen human bone

    Energy Technology Data Exchange (ETDEWEB)

    Antebi, Uri; Mathor, Monica B., E-mail: uri@usp.br, E-mail: mathor@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Guimaraes, Rodrigo P., E-mail: clinicaguimaraes@gmail.com [Santa Casa de Sao Paulo (FCM/SCSP), Sao Paulo, SP (Brazil). Faculdade de Ciencias Medicas

    2015-07-01

    The aim is the study of the application of ionizing radiation (gamma and electron) as sterilizing agents at doses of 15 kGy, 25 kGy and 50 kGy, the demineralized bone tissue frozen and freeze-dried for use in transplants. Five human femoral diaphysis of different donors demineralized bone tissues were preserved as lyophilized or frozen at - 80 deg C. The samples were divided into non-irradiated groups (control) and irradiated by gamma rays or electron beam. The bone proteins were extracted and used to determine the concentrations of total protein, BMP 2 and 7. It was observed a decrease in total protein concentrations, and BMP 2 and 7. The decrease in total protein concentrations, as compared to respective control groups was significant in the lyophilized and frozen samples irradiated at a dose of 50 kGy gamma radiation and beam electrons with greater than 30% reduction. The significant decrease in the levels of BMP 2 and 7 were also observed in higher doses and especially by electron beam. The reductions in the concentrations of total protein and osteoinductive proteins (BMP 2 and 7), were related to the radiation dose, i.e., increase with higher doses of ionizing radiation type and the type of preservation of the bones. The largest reductions in concentrations were observed in bone irradiated by electron beam and at a dose of 50 kGy. But this type of radiation and this high dose are not usual practice for the sterilization of bone tissue. Keywords: demineralized bone tissue, ionizing radiation, Tissue Bank, BMP 2, BMP 7, bone proteins. (author)

  2. Hepatoregenerative role of bone morphogenetic protein-9

    Science.gov (United States)

    Sosa, Ivan; Cvijanovic, Olga; Celic, Tanja; Cuculic, Drazen; Crncevic-Orlic, Zeljka; Vukelic, Lucian; Cvek, Sanja Zoricic; Dudaric, Luka; Bosnar, Alan; Bobinac, Dragica

    2011-01-01

    Summary Bone morphogenetic protein-9 (BMP-9) is a member of the transforming growth factor beta (TGF-β) superfamily of cytokines, which regulate cell growth and differentiation during embryogenesis. Apart of that, the hypoglycemic potential of BMP-9 is of great interest. It has been confirmed that BMP-9, like insulin, improves glycemia in diabetic mice and regulates directional glucose metabolism in hepatocytes; therefore it is proposed to be a candidate hepatic insulin-sensitizing substance (HISS). In liver fibrosis, due to the portocaval shunt, insulin bypasses the organ and the liver undergoes atrophy. Parenteral administration of insulin reverses atrophy by stimulating mitogenic activity of the hepatocytes. Because BMP-9 has a signaling pathway similar to other BMPs and insulin, it is to be expected that BMP-9 has a certain regenerative role in the liver, supporting the above-mentioned is evidence of BMP-9 expression in Dissè’s spaces and BMP-7’s mitogenic activity in mucosal cells. However, further studies are needed to confirm the possible regenerative role of BMP-9. PMID:22129908

  3. The history and histology of bone morphogenetic protein.

    Science.gov (United States)

    Murray, Samuel S; Brochmann Murray, Elsa J; Wang, Jeffrey C; Duarte, Maria Eugenia Leite

    2016-07-01

    Bone morphogenetic proteins are a group of structurally related proteins within the TGF-β superfamily of proteins with a diverse repertoire of functions in embryonic and adult organisms. As is apparent from the name, the members first characterized participate in bone growth, development, and remodeling. The "morphogenic" activity per se is defined as the induction of a recapitulation of endochondral bone formation by appropriate stem cells. The regenerative capacity of bone has been recognized since ancient times. The mechanism, applications, and conceptual basis of bone transplantation, bone implantation, ectopic bone formation, and exogenously induced bone formation have been studied by many investigators for more than a century. This review examines the efforts to characterize this activity in the European and American literature over approximately the last century. Because of the inherently complex nature of the process induced by these molecules (inflammation, stem cell proliferation, cartilage differentiation, replacement of cartilage with bone) it is important to evaluate previous investigations through a histological perspective. The cellular basis of the contemporary bioassay for BMP activity is illustrated and discussed from the histological point of view. PMID:26907674

  4. The Structure and Function of Non-Collagenous Bone Proteins

    Science.gov (United States)

    Hook, Magnus; McQuillan, David J.

    1997-01-01

    The research done under the cooperative research agreement for the project titled 'The structure and function of non-collagenous bone proteins' represented the first phase of an ongoing program to define the structural and functional relationships of the principal noncollagenous proteins in bone. An ultimate goal of this research is to enable design and execution of useful pharmacological compounds that will have a beneficial effect in treatment of osteoporosis, both land-based and induced by long-duration space travel. The goals of the now complete first phase were as follows: 1. Establish and/or develop powerful recombinant protein expression systems; 2. Develop and refine isolation and purification of recombinant proteins; 3. Express wild-type non-collagenous bone proteins; 4. Express site-specific mutant proteins and domains of wild-type proteins to enhance likelihood of crystal formation for subsequent solution of structure.

  5. Graphene oxide scaffold accelerates cellular proliferative response and alveolar bone healing of tooth extraction socket

    Science.gov (United States)

    Nishida, Erika; Miyaji, Hirofumi; Kato, Akihito; Takita, Hiroko; Iwanaga, Toshihiko; Momose, Takehito; Ogawa, Kosuke; Murakami, Shusuke; Sugaya, Tsutomu; Kawanami, Masamitsu

    2016-01-01

    Graphene oxide (GO) consisting of a carbon monolayer has been widely investigated for tissue engineering platforms because of its unique properties. For this study, we fabricated a GO-applied scaffold and assessed the cellular and tissue behaviors in the scaffold. A preclinical test was conducted to ascertain whether the GO scaffold promoted bone induction in dog tooth extraction sockets. For this study, GO scaffolds were prepared by coating the surface of a collagen sponge scaffold with 0.1 and 1 µg/mL GO dispersion. Scaffolds were characterized using scanning electron microscopy (SEM), physical testing, cell seeding, and rat subcutaneous implant testing. Then a GO scaffold was implanted into a dog tooth extraction socket. Histological observations were made at 2 weeks postsurgery. SEM observations show that GO attached to the surface of collagen scaffold struts. The GO scaffold exhibited an interconnected structure resembling that of control subjects. GO application improved the physical strength, enzyme resistance, and adsorption of calcium and proteins. Cytocompatibility tests showed that GO application significantly increased osteoblastic MC3T3-E1 cell proliferation. In addition, an assessment of rat subcutaneous tissue response revealed that implantation of 1 µg/mL GO scaffold stimulated cellular ingrowth behavior, suggesting that the GO scaffold exhibited good biocompatibility. The tissue ingrowth area and DNA contents of 1 µg/mL GO scaffold were, respectively, approximately 2.5-fold and 1.4-fold greater than those of the control. Particularly, the infiltration of ED2-positive (M2) macrophages and blood vessels were prominent in the GO scaffold. Dog bone-formation tests showed that 1 µg/mL GO scaffold implantation enhanced bone formation. New bone formation following GO scaffold implantation was enhanced fivefold compared to that in control subjects. These results suggest that GO was biocompatible and had high bone-formation capability for the scaffold

  6. Sentence Simplification Aids Protein-Protein Interaction Extraction

    OpenAIRE

    Jonnalagadda, Siddhartha; Gonzalez, Graciela

    2010-01-01

    Accurate systems for extracting Protein-Protein Interactions (PPIs) automatically from biomedical articles can help accelerate biomedical research. Biomedical Informatics researchers are collaborating to provide metaservices and advance the state-of-art in PPI extraction. One problem often neglected by current Natural Language Processing systems is the characteristic complexity of the sentences in biomedical literature. In this paper, we report on the impact that automatic simplification of s...

  7. Bone morphogenetic protein-2: a potential regulator in scleral remodeling

    OpenAIRE

    Hu, Jianmin; Cui, Dongmei; Yang, Xiao; Wang, Shaowei; Hu, Shoulong; Li, Chuanxu; Zeng, Junwen

    2008-01-01

    Purpose Bone morphogenetic protein 2 (BMP-2) is a member of the main subgroup of bone morphogenetic proteins within the transforming growth factor-β superfamily. BMP-2 is involved in numerous cellular functions including development, cell proliferation, apoptosis, and extracellular matrix synthesis. We examined BMP-2 expression in human scleral fibroblasts (HSF) and assessed the effects of recombinant human BMP-2 (rhBMP-2) on HSF proliferation, matrix metalloproteinase-2 (MMP-2), and tissue i...

  8. Sentence Simplification Aids Protein-Protein Interaction Extraction

    CERN Document Server

    Jonnalagadda, Siddhartha

    2010-01-01

    Accurate systems for extracting Protein-Protein Interactions (PPIs) automatically from biomedical articles can help accelerate biomedical research. Biomedical Informatics researchers are collaborating to provide metaservices and advance the state-of-art in PPI extraction. One problem often neglected by current Natural Language Processing systems is the characteristic complexity of the sentences in biomedical literature. In this paper, we report on the impact that automatic simplification of sentences has on the performance of a state-of-art PPI extraction system, showing a substantial improvement in recall (8%) when the sentence simplification method is applied, without significant impact to precision.

  9. Bone Flap Technique for Impacted Teeth Extraction and Bone Cysts Removal.

    Science.gov (United States)

    Saponaro, Gianmarco; Pelo, Sandro; De Angelis, Paolo; Forcione, Mario; D'Amato, Giuseppe; Moro, Alessandro

    2016-06-01

    The treatment of cystic lesions and the extraction of impacted third molars are 2 of the most common procedures in oral and maxillofacial surgery. The surgical treatment of cysts of the jaws can consist of a cystectomy, a cystotomy, or a staged combination of the 2 procedures. The surgical techniques developed for the extraction of impacted third molars are: coronectomy, orthodontic extraction, and surgery using intraoral or extraoral methods. There are various complications related to both surgical treatments. With regards to these complications, authors' department has developed a new surgical technique based on a previously described technique, which provides better support to the mucoperiosteal flap and improves bone regeneration after healing. Additionally, authors' goal was to reduce the risk of nerve injury, which has been achieved thanks to a direct visualization of the inferior alveolar nerve as well as cystic lesion or the dental element. The surgical procedure described produces major advantages over the traditional alternatives, despite needing a longer operation. This technique is particularly useful in the treatment of cystic lesions that have caused considerable bone loss. It can also be utilized for cysts or impacted dental elements strictly linked to the inferior alveolar nerve. PMID:27171951

  10. Modification of bone marrow radiosensitivity by medicinal plant extracts

    International Nuclear Information System (INIS)

    Withaferin A (WA), a steroidal lactone, and Plumbagin (Pi), a naphthoquinone, from the roots of Withania somnifera and Plumbage rosea, respectively, have been shows to possess growth inhibitory and radiosensitizing effects on experimental mouse tumours. An aqueous extract of the leaves of Ocimum sanctum (OE) was found to protect mice against radiation lethality. Therefore, the radiomodifying effects of the above plant products on the bone marrow of the adult Swiss mouse was studied. Single doses of WA (30 mg kg-1) or P1 (5 mg kg-1) were injected intraperitoneally tip) and OE (10 mg kg-1) was injected ip once daily for five consecutive days. Administration of extracts was followed by 2 Gy whole body gamma irradiation. Bone marrow stem cell survival was studied by an exogenous spleen colony unit (CFU-S) assay. The effects of WA and P1 were compared with that of cyclophosphamide (CP) and radioprotection by OE was compared with that of WR-2721 (WR). Radiation reduced the CFU-S to less than 50% of normal. WA, CP and P1 significantly enhanced this effect and reduced the CFU-S to almost the same extent (to <20% of normal), although individually WA and P1 were less cytotoxic than CP. These results indicate that radiosensitization by WE and P1 is not tumour specific. OE significantly increased CFU-S compared with radiotherapy (RT) alone. OE + RT gave a higher stem cell survival (p < 0.05) than that produced by WR + RT. While WR alone had a toxic effect, OE treatment showed no such effect, suggesting that the latter may have an advantage over WR in clinical application. (author)

  11. Expression of human bone-related proteins in the hematopoietic microenvironment.

    OpenAIRE

    Long, M W; Williams, J.L.; Mann, K G

    1990-01-01

    Given the intimate relationship between bone and bone marrow, we hypothesized that the human bone marrow may function as a source (or reservoir) of bone-forming progenitor cells. We observed a population of cells within the bone marrow which produce bone-specific or bone-related proteins. The production of these proteins was developmentally regulated in human long-term bone marrow cell cultures; the bone protein-producing cells (BPPC) are observed under serum-free, short-term culture conditio...

  12. TRANSCRIPTIONAL REGULATION OF BONE MARROW THROMBOPOIETIN BY PLATELET PROTEINS

    OpenAIRE

    McIntosh, Bryan; Kaushansky, Kenneth

    2008-01-01

    Platelet production is regulated primarily by the cytokine thrombopoietin (TPO). Although TPO is expressed in several different tissues, only in the bone marrow has the level of expression been reported to increase in response to reduced numbers of platelets. In these studies we demonstrate that platelet granule proteins are able to transcriptionally repress TPO mRNA expression in a marrow stromal cell line as well as in primary bone marrow stromal cell cultures. Like TPO mRNA, secretion of T...

  13. Inhibitory Effects of Morinda officinalis Extract on Bone Loss in Ovariectomized Rats

    OpenAIRE

    Qiao-Yan Zhang; Hong Zhang; Yan-Bin Wu; Ting Han; Lu-Ping Qin; Nan Li

    2009-01-01

    The present study was undertaken to investigate the protective effects of ethanol extract from the root of Morinda Officinalis (RMO) on ovariectomy-induced bone loss. Administration of RMO extract increased trabecular bone mineral content and bone mineral density of tibia, improved the levels of phosphorus (P), calcium (Ca) and OPG, decreased the levels of DPD/Cr, TRAP, ACTH and corticosterone, but did not reverse the levels of ALP, TNF-α and IL-6 in serum of ovariectomized rats. These findin...

  14. Graphene oxide scaffold accelerates cellular proliferative response and alveolar bone healing of tooth extraction socket

    Directory of Open Access Journals (Sweden)

    Nishida E

    2016-05-01

    Full Text Available Erika Nishida,1 Hirofumi Miyaji,1 Akihito Kato,1 Hiroko Takita,2 Toshihiko Iwanaga,3 Takehito Momose,1 Kosuke Ogawa,1 Shusuke Murakami,1 Tsutomu Sugaya,1 Masamitsu Kawanami11Department of Periodontology and Endodontology, Hokkaido University Graduate School of Dental Medicine, Sapporo, Japan; 2Support Section for Education and Research, Hokkaido University Graduate School of Dental Medicine, Sapporo, Japan; 3Laboratory of Histology and Cytology, Hokkaido University Graduate School of Medicine, Sapporo, JapanAbstract: Graphene oxide (GO consisting of a carbon monolayer has been widely investigated for tissue engineering platforms because of its unique properties. For this study, we fabricated a GO-applied scaffold and assessed the cellular and tissue behaviors in the scaffold. A preclinical test was conducted to ascertain whether the GO scaffold promoted bone induction in dog tooth extraction sockets. For this study, GO scaffolds were prepared by coating the surface of a collagen sponge scaffold with 0.1 and 1 µg/mL GO dispersion. Scaffolds were characterized using scanning electron microscopy (SEM, physical testing, cell seeding, and rat subcutaneous implant testing. Then a GO scaffold was implanted into a dog tooth extraction socket. Histological observations were made at 2 weeks postsurgery. SEM observations show that GO attached to the surface of collagen scaffold struts. The GO scaffold exhibited an interconnected structure resembling that of control subjects. GO application improved the physical strength, enzyme resistance, and adsorption of calcium and proteins. Cytocompatibility tests showed that GO application significantly increased osteoblastic MC3T3-E1 cell proliferation. In addition, an assessment of rat subcutaneous tissue response revealed that implantation of 1 µg/mL GO scaffold stimulated cellular ingrowth behavior, suggesting that the GO scaffold exhibited good biocompatibility. The tissue ingrowth area and DNA contents of 1

  15. Optimization of the Phenol -Chloroform Silica DNA Extraction Method in Ancient Bones DNA Extraction

    Directory of Open Access Journals (Sweden)

    Morteza Sadeghi

    2014-04-01

    Full Text Available Introduction: DNA extraction from the ancient bones tissues is currently very difficult. Phenol chloroform silica method is one of the methods currently used for this aim. The purpose of this study was to optimize the assessment method. Methods: DNA of 62 bone tissues (average 3-11 years was first extracted with phenol chloroform silica methods and then with changing of some parameters of the methods the extracted DNA was amplified in eight polymorphisms area including FES, F13, D13S317, D16, D5S818, vWA and CD4. Results from samples gained by two methods were compared in acrylamide gel. Results: The average of PCR yield for new method and common method in eight polymorphism regions was 75%, 78%, 81%, 76%, 85%, 71%, 89%, 86% and 64%, 39%, 70%, 49%, 68%, 76%, 71% and 28% respectively. The average of DNA in optimized (in 35l silica density and common method were 267.5 µg/ml with 1.12 purity and 192.76 g/ml with 0.84 purity respectively. Conclusions: According to the findings of this study, it is estimated that longer EDTA attendance is an efficient agent in removing calcium and also adequate density of silica particles can be efficient in removal of PCR inhibitors.

  16. /sup 99m/Tc-diphosphonate bone imaging and uptake in healing rat extraction sockets

    International Nuclear Information System (INIS)

    Clinically positive bone scans of the jaws may result from a variety of benign dental conditions. An experimental system for studying radionuclide imaging and uptake in the jaws of rats was developed. Sequential /sup 99m/Tc-diphosphonate bone scans and radionuclide uptake determinations were performed on rats after standardized extractions of their mandibular left first molars. Positive bone scans were seen 4 to 16 days after molar extraction, and increased radionuclide uptake was found in the healing extraction wounds 4 to 42 days after the extraction. Conventional radiography and histology fail to show unusual bony architecture in extraction sockets at such times. These results correlate with clinical findings in patients and suggest that human beings may have positive bone scans for several months after dental extraction

  17. Inhibitory effects of morinda officinalis extract on bone loss in ovariectomized rats.

    Science.gov (United States)

    Li, Nan; Qin, Lu-Ping; Han, Ting; Wu, Yan-Bin; Zhang, Qiao-Yan; Zhang, Hong

    2009-01-01

    The present study was undertaken to investigate the protective effects of ethanol extract from the root of Morinda Officinalis (RMO) on ovariectomy-induced bone loss. Administration of RMO extract increased trabecular bone mineral content and bone mineral density of tibia, improved the levels of phosphorus (P), calcium (Ca) and OPG, decreased the levels of DPD/Cr, TRAP, ACTH and corticosterone, but did not reverse the levels of ALP, TNF-alpha and IL-6 in serum of ovariectomized rats. These findings demonstrated that RMO extract reduced bone loss in ovariectomized rats, probably via the inhibition of bone resorption, but was not involved with bone formation. Anthraquinones and polysaccharides from Morinda officinals could be responsible for their antiosteoporotic activity, and the action mechanism of these constituents needs to be further studied. Therefore, RMO has the potential to develop a clinically useful antiosteoporotic agent. PMID:19513005

  18. Inhibitory Effects of Morinda officinalis Extract on Bone Loss in Ovariectomized Rats

    Directory of Open Access Journals (Sweden)

    Qiao-Yan Zhang

    2009-06-01

    Full Text Available The present study was undertaken to investigate the protective effects of ethanol extract from the root of Morinda Officinalis (RMO on ovariectomy-induced bone loss. Administration of RMO extract increased trabecular bone mineral content and bone mineral density of tibia, improved the levels of phosphorus (P, calcium (Ca and OPG, decreased the levels of DPD/Cr, TRAP, ACTH and corticosterone, but did not reverse the levels of ALP, TNF-α and IL-6 in serum of ovariectomized rats. These findings demonstrated that RMO extract reduced bone loss in ovariectomized rats, probably via the inhibition of bone resorption, but was not involved with bone formation. Anthraquinones and polysaccharides from Morinda officinals could be responsible for their antiosteoporotic activity, and the action mechanism of these constituents needs to be further studied. Therefore, RMO has the potential to develop a clinically useful antiosteoporotic agent.

  19. Proteomic analysis of a pleistocene mammoth femur reveals more than one hundred ancient bone proteins

    DEFF Research Database (Denmark)

    Cappellini, E.; Jensen, L.J.; Szklarczyk, D.; Ginolhac, A.; Da Fonseca, R.A.R.; Stafford, Thomas; Holen, S.R.; Collins, M.J.; Orlando, L.; Willerslev, E.; Gilbert, M Thomas P; Olsen, J.V.

    2012-01-01

    described beyond subpolar environments. Mass spectrometry-based ancient protein sequencing offers new perspectives for future molecular phylogenetic inference and physiological studies on samples not amenable to ancient DNA investigation. This approach therefore represents a further step into the ongoing......We used high-sensitivity, high-resolution tandem mass spectrometry to shotgun sequence ancient protein remains extracted from a 43 000 year old woolly mammoth (Mammuthus primigenius) bone preserved in the Siberian permafrost. For the first time, 126 unique protein accessions, mostly low...... evidence was observed of amino acid modifications due to post-mortem hydrolytic and oxidative damage. A consistent subset of this permafrost bone proteome was also identified in more recent Columbian mammoth (Mammuthus columbi) samples from temperate latitudes, extending the potential of the approach...

  20. Induction of Bone Matrix Protein Expression by Native Bone Matrix Proteins in C2C12 Culture

    Institute of Scientific and Technical Information of China (English)

    ZHEN-MING HU; SEAN A. F. PEEL; STEPHEN K. C. HO; GEORGE K. B. SANDOR; CAMERON M. L. CLOKIE

    2009-01-01

    Objective To study the expression of bone matrix protein (BMP) induced by bovine bone morphogenetic proteins (BMPs) in vitro. Methods Type I collagen, osteopontin (OPN), osteonectin (ON), osteocalcin (OC), and bone sialoprotein (BSP) were detected by immunohistochemistry in C2C12 cultured from day 1 to day 28. Results The signaling of bone matrix protein expression became weaker except for type I collagen, OC and BSP after 5 days. Fourteen days after culture, the positive signaling of type I collagen, OPN, ON, OC, and BSP was gradually declined, and could be detected significantly as compared with that of the negative control on day 28. BMP assay showed that the Ikaline phosphatase (ALP) activity was higher in C2C12 culture than in the control during the 14-day culture. Also, total protein and DNA significantly increased during the 14-day culture. High levels of ALP were seen in preosteoblasts and osteoblsts in vivo and in differentiating ostcoblasts in vitro. ALP was well recognized as a marker reflecting osteoblastic activity. Conclusion Native bovine BMP induces conversion of myoblasts into osteoblasts, produces type 1 collagen, and plays significantly role in osteoinduction and bone matrix mineralization of C2C12 in vitro.

  1. Effects of ionizing radiation on proteins in lyophilized or frozen demineralized human bone

    Science.gov (United States)

    Antebi, Uri; Mathor, Monica Beatriz; da Silva, André Ferreira; Guimarães, Rodrigo Pereira; Honda, Emerson Kiyoshi

    2016-01-01

    Objective The aim was to study the effects of application of ionizing radiation (gamma and electrons) as sterilizing agents at doses of 15 kGy, 25 kGy and 50 kGy, on lyophilized or frozen demineralized bone tissue for use in transplants. Methods Five human femoral diaphyses from different donors of musculoskeletal tissue were demineralized and preserved as lyophilized or frozen at −80 °C. The samples were divided into two groups: non-irradiated (control) and irradiated by means of gamma rays or an electron beam. The bone proteins were extracted and used to determine the concentrations of total protein and BMP 2 and 7. Results Decreases in total protein and BMP 2 and 7 concentrations were observed. The decreases in total protein concentrations, in comparison with the respective control groups, were significant in the lyophilized and frozen samples that were irradiated at a dose of 50 kGy of gamma radiation and electron beam, with reductions of more than 30%. Significant decreases in the levels of BMP 2 and 7 were also observed at higher doses and especially through use of the electron beam. Conclusion The reductions in the concentrations of total proteins and osteoinductive proteins (BMP 2 and 7) were related to the radiation dose, i.e. they increased with higher doses of ionizing radiation type and the type of bone preservation. The largest reductions in concentrations were observed in the bones irradiated by means of an electron beam and at a dose of 50 kGy. However, this type of radiation and this high dose are not usual practices for sterilization of bone tissue. PMID:27069893

  2. Yellow Mealworm Protein for Food Purposes - Extraction and Functional Properties

    OpenAIRE

    Zhao, Xue; Vázquez-Gutiérrez, José Luis; Johansson, Daniel P.; Landberg, Rikard; Langton, Maud

    2016-01-01

    A protocol for extraction of yellow mealworm larvae proteins was established, conditions were evaluated and the resulting protein extract was characterised. The freeze-dried yellow mealworm larvae contained around 33% fat, 51% crude protein and 43% true protein on a dry matter basis. The true protein content of the protein extract was about 75%, with an extraction rate of 70% under optimised extraction conditions using 0.25 M NaOH, a NaOH solution:ethanol defatted worm ratio of 15:1 mL/g, 40°...

  3. Use of bone morphogenetic proteins in mesenchymal stem cell stimulation of cartilage and bone repair

    OpenAIRE

    Scarfì, Sonia

    2016-01-01

    The extracellular matrix-associated bone morphogenetic proteins (BMPs) govern a plethora of biological processes. The BMPs are members of the transforming growth factor-β protein superfamily, and they actively participate to kidney development, digit and limb formation, angiogenesis, tissue fibrosis and tumor development. Since their discovery, they have attracted attention for their fascinating perspectives in the regenerative medicine and tissue engineering fields. BMPs have been employed i...

  4. Tracheal cartilage regeneration and new bone formation by slow release of bone morphogenetic protein (BMP)-2.

    Science.gov (United States)

    Igai, Hitoshi; Chang, Sung Soo; Gotoh, Masashi; Yamamoto, Yasumichi; Yamamoto, Masaya; Tabata, Yasuhiko; Yokomise, Hiroyasu

    2008-01-01

    We investigated the efficiency of bone morphogenetic protein (BMP)-2 released slowly from gelatin sponge for tracheal cartilage regeneration. A 1-cm gap was made in the mid-ventral portion of each of 10 consecutive tracheal cartilages. In the control group (n = 4), the resulting gap was left untreated. In the gelatin group (n = 4), plain gelatin was implanted in the gap. In the BMP-2 group (n = 4), gelatin containing 100 microg BMP-2 was implanted. We euthanatized all dogs in each group at 1, 3, 6, and 12 months after the implantation, respectively, and then examined the implant site macro- and microscopically. In the BMP-2 group, regenerated fibrous cartilage and newly formed bone were observed at 1 and 12 months. Regenerated cartilage was observed at the ends of the host cartilage stumps, with newly formed bone in the middle portion. The gaps were filled with regenerated cartilage and newly formed bone. At 3 and 6 months, regenerated cartilage, but not newly formed bone, was evident. The regenerated cartilage was covered with perichondrium and showed continuity with the host cartilage. We succeeded in inducing cartilage regeneration and new bone formation in canine trachea by slow release of 100 microg BMP-2 from gelatin. PMID:18204324

  5. Effect of the "protein diet" and bone tissue

    Directory of Open Access Journals (Sweden)

    Zoraide Nascimento da Silva

    2014-01-01

    Full Text Available The aim of this study is to evaluate the effect of the hyperproteic diet consumption on bone tissue. Methods: The study was conducted during sixty days. Twenty eight Wistar albinus rats, adults, originated from Laboratory of Experimental Nutrition were divided in four groups: (n = 7; Control 1 (C1, Control 2 (C2, Hyperproteic 1 (HP1 e Hyperproteic 2 (HP2. The C2 and HP2 groups were submitted to 30% of food restriction. The hyperproteic diet was based on the Atkins diet and prepared to simulate the protein diet. At the end of the study the animals were anesthetized to performer bone densitometry analyses by DEXA and blood and tissue collection. Serum and bone minerals analyses were conducted by colorimetric methods in automated equipment. Results: The total bone mineral density (BMD of the pelvis and the spine of the food restriction groups (HP2 e C2 were lower (p < 0.05 than C1 e HP1 groups. While the femur BMD of the HP2 was lower (p < 0.05 related to others groups. It had been observed reduction (p < 0.05 in the medium point of the width of femur diaphysis and in bone calcium level in the hyperproteic groups (HP1 e HP2. It was observed similar effect on the osteocalcin level, that presented lower (p < 0.05 in the hyperproteic groups. The insulin level was lower only in HP2 and serum calcium of the HP1 and HP2 groups was lower than C1. Conclusion: The protein diet promotes significant bone change on femur and in the hormones levels related to bone synthesis and maintenance of this tissue.

  6. Bone morphogenetic proteins: from structure to clinical use

    Directory of Open Access Journals (Sweden)

    Granjeiro J.M.

    2005-01-01

    Full Text Available Bone morphogenetic proteins (BMPs are multi-functional growth factors belonging to the transforming growth factor ß superfamily. Family members are expressed during limb development, endochondral ossification, early fracture, and cartilage repair. The activity of BMPs was first identified in the 1960s but the proteins responsible for bone induction were unknown until the purification and cloning of human BMPs in the 1980s. To date, about 15 BMP family members have been identified and characterized. The signal triggered by BMPs is transduced through serine/threonine kinase receptors, type I and II subtypes. Three type I receptors have been shown to bind BMP ligands, namely: type IA and IB BMP receptors and type IA activin receptors. BMPs seem to be involved in the regulation of cell proliferation, survival, differentiation and apoptosis, but their hallmark is their ability to induce bone, cartilage, ligament, and tendon formation at both heterotopic and orthotopic sites. This suggests that, in the future, they may play a major role in the treatment of bone diseases. Several animal studies have illustrated the potential of BMPs to enhance spinal fusion, repair critical-size defects, accelerate union, and heal articular cartilage lesions. Difficulties in producing and purifying BMPs from bone tissue have prompted the attempts made by several laboratories, including ours, to express these proteins in the recombinant form in heterologous systems. This review focuses on BMP structure, molecular mechanisms of action and significance and potential applications in medical, dental and veterinary practice for the treatment of cartilage and bone-related diseases.

  7. Up-regulation of bone marrow stromal protein 2 (BST2) in breast cancer with bone metastasis

    OpenAIRE

    Zheng Xin; Li Zhen; Cao Jie; Cai Dongqing; Yao Yao; Li Wanglin; Yuan Ziqiang

    2009-01-01

    Abstract Background Bone metastases are frequent complications of breast cancer. Recent literature implicates multiple chemokines in the formation of bone metastases in breast cancer. However, the molecular mechanism of metastatic bone disease in breast cancer remains unknown. We have recently made the novel observation of the BST2 protein expression in human breast cancer cell lines. The purpose of our present study is to investigate the expression and the role of BST2 in bone metastatic bre...

  8. PROTEIN OF MEAT AND BONE MEAL FOR PIGS

    Directory of Open Access Journals (Sweden)

    Patieva S. V.

    2015-09-01

    Full Text Available The modern requirements of intergovernmental standards to the quality and safety of livestock produce provide for the use of highly productive animals capable under small expenses to produce more the high quality produce. In particular, at the formation of meat productivity at pigs the great significance has an achievement of optimal digestion and assimilability of consumed fodder means. In the connection, the study of digestion of meat and bone meal from slaughterhouse wastes of cattle (MCM and poultry (MCBM presents the scientific interest. In the fodder experience on the growing pigs with the fistula of iliac intestines there was investigated the digestion of two types of meat and bone meal from slaughterhouse wastes of cattle (MCM and poultry (MKBM. The iliac accessibility of amino acids of meat and bone meal found itself too low: 49,3 % - 69,3 %. The accessibility of general protein reliably did not differ from the average accessibility on main amino acids - 61,5 %. To count the real iliac accessibility of raw protein and amino acids of meat and bone meal there was determined an endogenous emission of these substances on the casein diet. The real iliac accessibility of protein and individual amino acids did not leave the limits in 73% on МCM and 69% - on МCBМ. The accessibility of lysine, leucine and isoleucine MCBM is reliably higher than the same in MCM (P

  9. Comparison Between Gelatines Extracted From Mackerel and Blue Whiting Bones after Different Pre-treatments

    OpenAIRE

    Barry-Ryan, Catherine; Khiari, Zied; Rico, Daniel; Martin-Diana, Ana Belen

    2013-01-01

    Gelatines were extracted from mackerel and blue whiting bones after chemical or enzymatic pre-treatments and their functional properties (solubility, foaming and emulsifying properties) were analysed. The pre-treatment significantly (p

  10. Whey Protein Concentrate Hydrolysate Prevents Bone Loss in Ovariectomized Rats.

    Science.gov (United States)

    Kim, Jonggun; Kim, Hyung Kwan; Kim, Saehun; Imm, Ji-Young; Whang, Kwang-Youn

    2015-12-01

    Milk is known as a safe food and contains easily absorbable minerals and proteins, including whey protein, which has demonstrated antiosteoporotic effects on ovariectomized rats. This study evaluated the antiosteoporotic effect of whey protein concentrate hydrolysate (WPCH) digested with fungal protease and whey protein concentrate (WPC). Two experiments were conducted to determine (1) efficacy of WPCH and WPC and (2) dose-dependent impact of WPCH in ovariectomized rats (10 weeks old). In Experiment I, ovariectomized rats (n=45) were allotted into three dietary treatments of 10 g/kg diet of WPC, 10 g/kg diet of WPCH, and a control diet. In Experiment II, ovariectomized rats (n=60) were fed four different diets (0, 10, 20, and 40 g/kg of WPCH). In both experiments, sham-operated rats (n=15) were also fed a control diet containing the same amount of amino acids and minerals as dietary treatments. After 6 weeks, dietary WPCH prevented loss of bone, physical properties, mineral density, and mineral content, and improved breaking strength of femurs, with similar effect to WPC. The bone resorption enzyme activity (tartrate resistance acid phosphatase) in tibia epiphysis decreased in response to WPCH supplementation, while bone formation enzyme activity (alkaline phosphatase) was unaffected by ovariectomy and dietary treatment. Bone properties and strength increased as the dietary WPCH level increased (10 and 20 g/kg), but there was no difference between the 20 and 40 g/kg treatment. WPCH and WPC supplementation ameliorated bone loss induced by ovariectomy in rats. PMID:26367331

  11. A novel method of isolation of a bone-morphogenetic-protein-like protein from ossein.

    Science.gov (United States)

    Mythili, J; Padmavathy, S; Chandrakasan, G

    2001-08-01

    A new bone-morphogenetic-protein (BMP)-like protein has been isolated through a new protocol from a novel source, ossein. The BMP-like protein was hydrophilic and characterized through Fourier-transform IR studies, SDS/PAGE and coupled with a neutral binder, hydroxypropylmethylcellulose (HPMC) for control release. The IR spectrum of the protein showed peaks in tandem with BMP from bone matrix, and its molecular mass was in the range 18-21 kDa. Sustained release from the surface of HPMC was achieved for a period of 3 days. PMID:11483152

  12. Comparison of three methods of DNA extraction from human bones with different degrees of degradation

    OpenAIRE

    Jakubowska, Joanna; Maciejewska, Agnieszka; Pawłowski, Ryszard

    2011-01-01

    There is a necessity for deceased identification as a result of many accidents and sometimes bones are the only accessible source of DNA. So far, a universal method that allows for extraction of DNA from materials at different stages of degradation does not exist. The aims of this study were: the comparison of three methods of DNA extraction from bones with different degree of degradation and an evaluation of the usefulness of these methods in forensic genetics. The efficiency of DNA extracti...

  13. AMP-activated protein kinase (AMPK) activation regulates in vitro bone formation and bone mass.

    Science.gov (United States)

    Shah, M; Kola, B; Bataveljic, A; Arnett, T R; Viollet, B; Saxon, L; Korbonits, M; Chenu, C

    2010-08-01

    Adenosine 5'-monophosphate-activated protein kinase (AMPK), a regulator of energy homeostasis, has a central role in mediating the appetite-modulating and metabolic effects of many hormones and antidiabetic drugs metformin and glitazones. The objective of this study was to determine if AMPK can be activated in osteoblasts by known AMPK modulators and if AMPK activity is involved in osteoblast function in vitro and regulation of bone mass in vivo. ROS 17/2.8 rat osteoblast-like cells were cultured in the presence of AMPK activators (AICAR and metformin), AMPK inhibitor (compound C), the gastric peptide hormone ghrelin and the beta-adrenergic blocker propranolol. AMPK activity was measured in cell lysates by a functional kinase assay and AMPK protein phosphorylation was studied by Western Blotting using an antibody recognizing AMPK Thr-172 residue. We demonstrated that treatment of ROS 17/2.8 cells with AICAR and metformin stimulates Thr-172 phosphorylation of AMPK and dose-dependently increases its activity. In contrast, treatment of ROS 17/2.8 cells with compound C inhibited AMPK phosphorylation. Ghrelin and propranolol dose-dependently increased AMPK phosphorylation and activity. Cell proliferation and alkaline phosphatase activity were not affected by metformin treatment while AICAR significantly inhibited ROS 17/2.8 cell proliferation and alkaline phosphatase activity at high concentrations. To study the effect of AMPK activation on bone formation in vitro, primary osteoblasts obtained from rat calvaria were cultured for 14-17days in the presence of AICAR, metformin and compound C. Formation of 'trabecular-shaped' bone nodules was evaluated following alizarin red staining. We demonstrated that both AICAR and metformin dose-dependently increase trabecular bone nodule formation, while compound C inhibits bone formation. When primary osteoblasts were co-treated with AICAR and compound C, compound C suppressed the stimulatory effect of AICAR on bone nodule formation

  14. Human Cementum Protein 1 induces expression of bone and cementum proteins by human gingival fibroblasts

    International Nuclear Information System (INIS)

    We recently presented evidence showing that a human cementoblastoma-derived protein, named Cementum Protein 1 (CEMP1) may play a role as a local regulator of cementoblast differentiation and cementum-matrix mineralization. This protein was shown to be expressed by cementoblasts and progenitor cells localized in the periodontal ligament. In this study we demonstrate that transfection of CEMP1 into human gingival fibroblasts (HGF) induces mineralization and expression of bone and cementum-matrix proteins. The transfected HGF cells had higher alkaline phosphatase activity and proliferation rate and they expressed genes for alkaline phosphatase, bone sialoprotein, osteocalcin, osteopontin, the transcription factor Runx2/Cbfa1, and cementum attachment protein (CAP). They also produced biological-type hydroxyapatite. These findings indicate that the CEMP1 might participate in differentiation and mineralization of nonosteogenic cells, and that it might have a potential function in cementum and bone formation

  15. Study on osteopotential activity of Terminalia arjuna bark extract incorporated bone substitute

    Indian Academy of Sciences (India)

    G Krithiga; T Hemalatha; R Deepachitra; Kausik Ghosh; T P Sastry

    2014-10-01

    Bark extract of Terminalia arjuna (TA) possesses potent medical properties and therefore, holds a reputed position in both Ayurvedic and Unani systems of medicine. Bone substitutes play an inevitable role in traumatic bone damages. Growth factors induce osteoinductivity, but suffer from limitations such as high cost and side effects. This study aims to evaluate the osteoinductive potential of bark extract of TA in bone substitutes. Bone substitutes prepared with TA bark extract were characterized for their physicochemical properties. In vitro biomineralization study was carried out using simulated body fluid. Cytotoxicity, alkaline phosphatase activity and mineralization potential were assessed using MG-63 cell lines. Scanning electron microscope revealed apatite formation on the surface after biomineralization. Thermogravimetric analysis showed 15% increase in residual weight by deposition of calcium and phosphate and their presence was identified by energy dispersive analysis. Increased alkaline phosphatase and calcium release was observed in bone substitutes prepared with TA extract compared with control. The functional groups of TA bark extract help in in vitro biomineralization. In MG-63 cells, it showed potential influence in cell differentiation. TA extract may be used as low-cost alternative for growth factors for treatment of fractured bones.

  16. Ectopic bone induction in porous apatite-wollastonite-containing glass ceramic combined with bone morphogenetic protein.

    Science.gov (United States)

    Ijiri, S; Nakamura, T; Fujisawa, Y; Hazama, M; Komatsudani, S

    1997-06-15

    To accelerate the integration of ceramic implants with the surrounding bone and to search for a suitable carrier for bone morphogenetic protein (BMP), we studied ectopic bone induction in porous apatite-wollastonite-containing glass ceramic (A-W GC) combined with partially purified bovine BMP (bBMP) and recombinant Xenopus BMP-4/7 (rxBMP-4/7). Porous A-W GC rods [4 mm in diameter, 5 mm in height, 70% porosity, 200 microns mean pore size, 17.54 +/- 3.82 MPa (mean +/- SD) compressive strength] were used. An apatite coating formed on the surface of porous A-W GC that had been immersed in simulated body fluid at 36.5 degrees C for 7 days. In experiment 1, porous A-W GC rods were combined with either bBMP, collagen, or with both bBMP and collagen. The rods were implanted into subcutaneous pouches in rats and were harvested 4 weeks after implantation. Low-energy radiographic, scanning electron microscopic (SEM), and histological examinations showed ectopic bone formation and within the rods only in the porous A-W GC combined with the bBMP and collagen group. Quantitative analysis also revealed that this group alone showed a significant increase in bone formation. In experiment 2, porous A-W GC rods were combined with rxBMP and collagen, implanted into rats, and harvested as described above. SEM and histological examination showed ectopic bone formation around and within the rods. Because of its relatively high mechanical strength, ease of handling, and good osteoinductivity, porous A-W GC combined with BMP and collagen may be clinically useful in patients with large cancellous bone defects or craniomaxillofacial lesions. PMID:9189820

  17. Multi-protein delivery by nanodiamonds promotes bone formation.

    Science.gov (United States)

    Moore, L; Gatica, M; Kim, H; Osawa, E; Ho, D

    2013-11-01

    Bone morphogenetic proteins (BMPs) are well-studied regulators of cartilage and bone development that have been Food and Drug Administration (FDA)-approved for the promotion of bone formation in certain procedures. BMPs are seeing more use in oral and maxillofacial surgeries because of recent FDA approval of InFUSE(®) for sinus augmentation and localized alveolar ridge augmentation. However, the utility of BMPs in medical and dental applications is limited by the delivery method. Currently, BMPs are delivered to the surgical site by the implantation of bulky collagen sponges. Here we evaluate the potential of detonation nanodiamonds (NDs) as a delivery vehicle for BMP-2 and basic fibroblast growth factor (bFGF). Nanodiamonds are biocompatible, 4- to 5-nm carbon nanoparticles that have previously been used to deliver a wide variety of molecules, including proteins and peptides. We find that both BMP-2 and bFGF are readily loaded onto NDs by physisorption, forming a stable colloidal solution, and are triggered to release in slightly acidic conditions. Simultaneous delivery of BMP-2 and bFGF by ND induces differentiation and proliferation in osteoblast progenitor cells. Overall, we find that NDs provide an effective injectable alternative for the delivery of BMP-2 and bFGF to promote bone formation. PMID:24045646

  18. Harmine promotes osteoblast differentiation through bone morphogenetic protein signaling

    International Nuclear Information System (INIS)

    Highlights: → Harmine promotes the activity and mRNA expression of ALP. → Harmine enhances the expressions of osteocalcin mRNA and protein. → Harmine induces osteoblastic mineralization. → Harmine upregulates the mRNA expressions of BMPs, Runx2 and Osterix. → BMP signaling pathways are involved in the actions of harmine. -- Abstract: Bone mass is regulated by osteoblast-mediated bone formation and osteoclast-mediated bone resorption. We previously reported that harmine, a β-carboline alkaloid, inhibits osteoclast differentiation and bone resorption in vitro and in vivo. In this study, we investigated the effects of harmine on osteoblast proliferation, differentiation and mineralization. Harmine promoted alkaline phosphatase (ALP) activity in MC3T3-E1 cells without affecting their proliferation. Harmine also increased the mRNA expressions of the osteoblast marker genes ALP and Osteocalcin. Furthermore, the mineralization of MC3T3-E1 cells was enhanced by treatment with harmine. Harmine also induced osteoblast differentiation in primary calvarial osteoblasts and mesenchymal stem cell line C3H10T1/2 cells. Structure-activity relationship studies using harmine-related β-carboline alkaloids revealed that the C3-C4 double bond and 7-hydroxy or 7-methoxy group of harmine were important for its osteogenic activity. The bone morphogenetic protein (BMP) antagonist noggin and its receptor kinase inhibitors dorsomorphin and LDN-193189 attenuated harmine-promoted ALP activity. In addition, harmine increased the mRNA expressions of Bmp-2, Bmp-4, Bmp-6, Bmp-7 and its target gene Id1. Harmine also enhanced the mRNA expressions of Runx2 and Osterix, which are key transcription factors in osteoblast differentiation. Furthermore, BMP-responsive and Runx2-responsive reporters were activated by harmine treatment. Taken together, these results indicate that harmine enhances osteoblast differentiation probably by inducing the expressions of BMPs and activating BMP and Runx2

  19. Optimizing bone health in older adults: the importance of dietary protein

    OpenAIRE

    Surdykowski, Anna K; Kenny, Anne M.; Insogna, Karl L.; Kerstetter, Jane E

    2010-01-01

    Age-related bone loss is progressive and can lead to osteoporosis. While it is accepted that both dietary calcium and vitamin D are important and beneficial for skeletal health, the impact of dietary protein on calcium metabolism and bone balance remains controversial. Contrary to the hypothesis that increasing dietary protein contributes to bone loss, research supports the notion that protein may play a pivotal role in maintenance of bone health by several mechanisms; for example, increasing...

  20. Bone Morphogenetic Protein 4 Mediates Human Embryonic Germ Cell Derivation

    OpenAIRE

    Hiller, Marc; Liu, Cyndi; Blumenthal, Paul D; John D Gearhart; Kerr, Candace L.

    2010-01-01

    Human primordial germ cells (PGCs) have proven to be a source of pluripotent stem cells called embryonic germ cells (EGCs). Unlike embryonic stem cells, virtually little is known regarding the factors that regulate EGC survival and maintenance. In mice, the growth factor bone morphogenetic protein 4 (BMP4) has been shown to be required for maintaining mouse embryonic stem cells, and disruptions in this gene lead to defects in mouse PGC specification. Here, we sought to determine whether recom...

  1. Automatic bone segmentation and bone-cartilage interface extraction for the shoulder joint from magnetic resonance images

    International Nuclear Information System (INIS)

    We present a statistical shape model approach for automated segmentation of the proximal humerus and scapula with subsequent bone-cartilage interface (BCI) extraction from 3D magnetic resonance (MR) images of the shoulder region. Manual and automated bone segmentations from shoulder MR examinations from 25 healthy subjects acquired using steady-state free precession sequences were compared with the Dice similarity coefficient (DSC). The mean DSC scores between the manual and automated segmentations of the humerus and scapula bone volumes surrounding the BCI region were 0.926  ±  0.050 and 0.837  ±  0.059, respectively. The mean DSC values obtained for BCI extraction were 0.806  ±  0.133 for the humerus and 0.795  ±  0.117 for the scapula. The current model-based approach successfully provided automated bone segmentation and BCI extraction from MR images of the shoulder. In future work, this framework appears to provide a promising avenue for automated segmentation and quantitative analysis of cartilage in the glenohumeral joint. (paper)

  2. Extractable proteins from irradiated field natural rubber latex

    Energy Technology Data Exchange (ETDEWEB)

    Rogero, S.O.; Lugao, A.B.; Yoshii, Fumio; Makuuchi, Keizo

    2003-06-01

    In this study field natural rubber latex was irradiated with different doses near a {sup 60}Co gamma source to reduce the water-soluble protein content in the final product. The protein content of the films obtained by casting method was extracted with phosphate buffer solution, pH 7 and was measured using Micro BCA Protein Assay kit. Also was measured protein in the serum samples of field NRL. The concentration of extractable proteins increased with increasing radiation dose.

  3. Extractable proteins from irradiated field natural rubber latex

    International Nuclear Information System (INIS)

    In this study field natural rubber latex was irradiated with different doses near a 60Co gamma source to reduce the water-soluble protein content in the final product. The protein content of the films obtained by casting method was extracted with phosphate buffer solution, pH 7 and was measured using Micro BCA Protein Assay kit. Also was measured protein in the serum samples of field NRL. The concentration of extractable proteins increased with increasing radiation dose

  4. Extractable proteins from irradiated field natural rubber latex

    Science.gov (United States)

    Rogero, Sizue O.; Lugão, Ademar B.; Yoshii, Fumio; Makuuchi, Keizo

    2003-06-01

    In this study field natural rubber latex was irradiated with different doses near a 60Co gamma source to reduce the water-soluble protein content in the final product. The protein content of the films obtained by casting method was extracted with phosphate buffer solution, pH 7 and was measured using Micro BCA Protein Assay kit. Also was measured protein in the serum samples of field NRL. The concentration of extractable proteins increased with increasing radiation dose.

  5. The Structure and Function of Non-Collagenous Bone Proteins

    Science.gov (United States)

    Hook, Magnus

    1997-01-01

    The long-term goal for this program is to determine the structural and functional relationships of bone proteins and proteins that interact with bone. This information will used to design useful pharmacological compounds that will have a beneficial effect in osteoporotic patients and in the osteoporotic-like effects experienced on long duration space missions. The first phase of this program, funded under a cooperative research agreement with NASA through the Texas Medical Center, aimed to develop powerful recombinant expression systems and purification methods for production of large amounts of target proteins. Proteins expressed in sufficient'amount and purity would be characterized by a variety of structural methods, and made available for crystallization studies. In order to increase the likelihood of crystallization and subsequent high resolution solution of structures, we undertook to develop expression of normal and mutant forms of proteins by bacterial and mammalian cells. In addition to the main goals of this program, we would also be able to provide reagents for other related studies, including development of anti-fibrotic and anti-metastatic therapeutics.

  6. Effect of biomaterial properties on bone healing in a rabbit tooth extraction socket model

    NARCIS (Netherlands)

    Fisher, J.P.; Lalani, Z.; Bossano, C.M.; Brey, E.M.; Demian, N.; Johnston, C.M.; Dean, D.; Jansen, J.A.; Wong, M.E.; Mikos, A.G.

    2004-01-01

    In this work we sought to understand the effect of biomaterial properties upon healing bone tissue. We hypothesized that a hydrophilic polymer gel implanted into a bone tissue defect would impede the healing process owing to the biomaterial's prevention of protein adsorption and thus cell adhesion.

  7. Production of surgical gloves from low extractable protein RVNRL

    Energy Technology Data Exchange (ETDEWEB)

    Marga, Utama; Yanti, S.; Made, Sumarti; Marsongko; Tita, Puspitasari; Dian, Iramani [Center for Research and Development of Isotopes and Radiation Technology, National Nuclear Energy Agency, Jakarta (Indonesia); Makuuchi, K. [EB System Cooperation, Takasaki, Gunma (Japan); Yoshii, F. [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment; Siswanto [Research Unit for Biotechnology of Estate Crop (Indonesia)

    2001-03-01

    Study on the production of surgical gloves from low extractable protein PVNRL (Radiation Vulcanization of Natural Rubber Latex) in home industry scale with normal butyl acrylate as sensitizer has been carried out. The variation of dipping speed, concentration of coagulant agent and selection of antioxidant for producing good quality of surgical gloves were evaluated. The water-extractable protein and PBS (Phosphate Buffer Saline) - extractable protein content, the physical and mechanical properties of gloves were measured. The results show that for producing a good quality of surgical gloves from low extractable protein RVNRL, the concentration of latex is 50% with calcium nitrate as coagulant agent between 15-20%. By using this condition the physical and mechanical properties of surgical gloves is required to ASTM standard such as tensile strength more than 24 MPa, PBS-extractable protein is around 41-68 ug/g and water-extractable protein contents is around 23-35 ug/g. (author)

  8. Production of surgical gloves from low extractable protein RVNRL

    International Nuclear Information System (INIS)

    Study on the production of surgical gloves from low extractable protein PVNRL (Radiation Vulcanization of Natural Rubber Latex) in home industry scale with normal butyl acrylate as sensitizer has been carried out. The variation of dipping speed, concentration of coagulant agent and selection of antioxidant for producing good quality of surgical gloves were evaluated. The water-extractable protein and PBS (Phosphate Buffer Saline) - extractable protein content, the physical and mechanical properties of gloves were measured. The results show that for producing a good quality of surgical gloves from low extractable protein RVNRL, the concentration of latex is 50% with calcium nitrate as coagulant agent between 15-20%. By using this condition the physical and mechanical properties of surgical gloves is required to ASTM standard such as tensile strength more than 24 MPa, PBS-extractable protein is around 41-68 ug/g and water-extractable protein contents is around 23-35 ug/g. (author)

  9. A fucoidan from Nemacystus decipiens disrupts angiogenesis through targeting bone morphogenetic protein 4.

    Science.gov (United States)

    Wang, Wucheng; Chen, Huanjun; Zhang, Lei; Qin, Yi; Cong, Qifei; Wang, Peipei; Ding, Kan

    2016-06-25

    A sulfated and acetylated fucoidan, named NDH01, was extracted from seaweed Nemacystus decipiens. NDH01 was composed of mannose, glucuronic acid, fucose, sulfate group and acetyl group in the molar ratio of 3.0: 14.4: 82.6: 34.3: 13.9. The backbone of NDH01 was fucose-free core, composed of α-d-1,2-Manp and β-d-1,4-GlcpA disaccharide repeat unit. The branches were attached at the C3, C4 and C6 of α-d-1,2-Manp. The sidechain was composed of α-l-1,3,4-Fucp, α-l-1,4-Fucp, α-l-1,3-Fucp and α-l-1,4-GlcpA. The sulfate group was linked to C4 of α-l-1,3,4-Fucp, whereas, acetyl group was branched on C2 of α-l-1,2,3-Fucp. NDH01 could disrupt tube formation and inhibit the migration as well as cell growth of human microvascular endothelial cells. Besides, phosphorylation of Smad/1/5/8, Erk and FAK was significantly inhibited by NDH01. Further studies uncovered that NDH01 blocked Smad1/5/8 signaling via interacting with bone morphogenetic protein 4 and downregulating bone morphogenetic protein 4 expression. The results suggested that NDH01 might be an angiogenesis inhibitor through targeting bone morphogenetic protein 4. PMID:27083822

  10. Imaging symptomatic bone morphogenetic protein-2-induced heterotopic bone formation within the spinal canal: case report.

    Science.gov (United States)

    Chryssikos, Timothy; Crandall, Kenneth M; Sansur, Charles A

    2016-05-01

    Heterotopic bone formation within the spinal canal is a known complication of bone morphogenetic protein-2 (BMP-2) and presents a clinical and surgical challenge. Imaging modalities are routinely used for operative planning in this setting. Here, the authors present the case of a 59-year-old woman with cauda equina syndrome following intraoperative BMP-2 administration. Plain film myelographic studies showed a region of severe stenosis that was underappreciated on CT myelography due to a heterotopic bony lesion mimicking the dorsal aspect of a circumferentially patent thecal sac. When evaluating spinal stenosis under these circumstances, it is important to carefully consider plain myelographic images in addition to postmyelography CT images as the latter may underestimate the true degree of stenosis due to the potentially similar radiographic appearances of evolving BMP-2-induced heterotopic bone and intrathecal contrast. Alternatively, comparison of sequentially acquired noncontrast CT scans with CT myelographic images may also assist in distinguishing BMP-2-induced heterotopic bony lesions from the thecal sac. Further studies are needed to elucidate the roles of the available imaging techniques in this setting and to characterize the connection between the radiographic and histological appearances of BMP-2-induced heterotopic bone. PMID:26824586

  11. Quick and effective method of bone marrow mesenchymal stem cell extraction

    Directory of Open Access Journals (Sweden)

    Gudleviciene Zivile

    2015-01-01

    Full Text Available Background. Mesenchymal stem cells (MSCs are currently exploited in numerous clinical trials to investigate their potential in immune regulation, hematopoesis or tissue regeneration. The most common source of MSCs for clinical use is human bone marrow. To generate sufficient numbers of cells relevant to clinical use in most cases the high volumes (20-50 ml of bone marrow aspirates are taken. Methods. In this pilot study, 8 healthy bone marrow donors were included. Two different MSC extraction methods were evaluated: MSCs extraction from 60 ml of bone marrow using density gradient and MSCs extraction from 6 ml using red blood cell (RBC lysis. Results. Our results showed that after RBC lysis the efficient amount of human MSCs can be isolated from 10 times less bone marrow volume (6 ml. Moreover, using small volume of bone marrow the adequate therapeutical dose of MSCs could be achieved during similar period of time (3-4 weeks. In conclusion, we have shown that MSCs isolation using RBC lysis is an effective and more advantageous method in comparison to standard MSCs isolation using density-gradient. Using RBC lysis from small volume of bone marrow the same amount of MSCs were obtained as usually using large volume and density-gradient.

  12. Anthelmintic activity of Leucaena leucocephala protein extracts on Haemonchus contortus.

    Science.gov (United States)

    Soares, Alexandra Martins dos Santos; de Araújo, Sandra Alves; Lopes, Suzana Gomes; Costa Junior, Livio Martins

    2015-01-01

    The objective of this study was to evaluate the effects of protein extracts obtained from the plant Leucaena leucocephala on the nematode parasite Haemonchus contortus. The seeds, shell and cotyledon of L. leucocephala were separated and their proteins extracted using a sodium phosphate buffer, and named as TE (total seed extract), SE (shell extract) and CE (cotyledon extract). Soluble protein content, protease, protease inhibitory and chitinase activity assays were performed. Exsheathment inhibition of H. contortus larvae were performed at concentrations of 0.6 mg mL-1, and egg hatch assays were conducted at protein concentrations of 0.8, 0.4, 0.2, 0.1 and 0.05 mg mL-1. The effective concentration for 50% hatching inhibition (EC50) was estimated by probit. Different proportions of soluble proteins, protease and chitinase were found in TE and CE. Protease inhibitory activity was detected in all extracts. The EC50 of the CE and TE extracts were 0.48 and 0.33 mg mL-1, respectively. No ovicidal effects on H. contortus were detected in SE extracts, and none of the protein extracts demonstrated larvicidal effects on H. contortus. We therefore conclude that protein extracts of L. leucocephala had a detrimental effect on nematode eggs, which can be correlated with the high protease and chitinase activity of these extracts. PMID:26689178

  13. Methanol Extract of the Fruits of Morinda citrifolia Linn., Restores Bone Loss in Ovariectomized Rats

    Directory of Open Access Journals (Sweden)

    V. Parmar

    2011-01-01

    Full Text Available The objective of this study was to evaluate the effect of methanol extract of the fruits of Morinda citrifolia Linn., on osteoporosis induced by ovariectomy in female albino rats at two different dose levels of 500 and 750 mg/kg/day. Healthy female albino rats in the age group of 90 days were selected and randomized into five groups of six animals each. Group 1 was sham operated and served as control while all the remaining groups were ovariectomized. Group 2 was fed with an equivolume of saline and served as ovariectomized control. Group 3 was orally treated with standard Raloxifene (5.4 mg kg-1 whereas the methanol extract of Morinda citrifolia (500 and 750 mg kg-1 was administered to the groups 4 and 5. The findings assessed on the basis of biomechanical, biochemical and histopathological parameters, showed that the methanol extract significantly reduced bone loss, as evidenced by a reduction in Tartrate Resistant Acid Phosphatase (TRAP and urine Hydroxyproline (Hp levels while simultaneously increasing bone formation [high serum Alkaline Phosphatase (ALP levels], thereby restoring bone mineralization. The restoration of bone strength was confirmed by biomechanical parameters viz., the three point bending of tibia, load testing of femoral head and compression of IV lumbar vertebra and it was further endorsed by histopathological findings i.e., bone microarchitecture. The extract significantly increased the osteoblastic activity on one hand while on the other it retarded the osteoclastic function thereby contributing to a positive bone balance and hence enhanced mineralization.

  14. Intramembranous Bone Healing Process Subsequent to Tooth Extraction in Mice: Micro-Computed Tomography, Histomorphometric and Molecular Characterization

    OpenAIRE

    Vieira, Andreia Espindola; Repeke, Carlos Eduardo; Ferreira Junior, Samuel de Barros; Colavite, Priscila Maria; Biguetti, Claudia Cristina; de Oliveira, Rodrigo Cardoso; ASSIS, Gerson Francisco; Taga, Rumio; Trombone, Ana Paula Favaro; Garlet, Gustavo Pompermaier

    2015-01-01

    Bone tissue has a significant potential for healing, which involves a significant the interplay between bone and immune cells. While fracture healing represents a useful model to investigate endochondral bone healing, intramembranous bone healing models are yet to be developed and characterized. In this study, a micro-computed tomography, histomorphometric and molecular (RealTimePCRarray) characterization of post tooth-extraction alveolar bone healing was performed on C57Bl/6 WT mice. After t...

  15. Successful treatment of a humeral capitulum osteonecrosis with bone morphogenetic protein-7 combined with autologous bone grafting

    OpenAIRE

    Marsell, Richard; Hailer, Nils P

    2014-01-01

    We present the case of a 27-year-old female with subcortical osteonecrosis of the humeral capitulum. Percutaneous retrograde drilling of the lesion and application of recombinant human bone morphogenetic protein (BMP)-7 were combined with autologous bone grafting. At follow-up the patient was almost pain-free, had normalized her range of motion, and radiography showed consolidation of the lesion without any heterotopic bone formation. By timing surgery prior to subchondral collapse, biomechan...

  16. Calcium Phosphate Scaffolds Combined with Bone Morphogenetic Proteins or Mesenchymal Stem Cells in Bone Tissue Engineering

    Directory of Open Access Journals (Sweden)

    Han Sun

    2015-01-01

    Full Text Available Objective: The purpose of this study was to review the current status of calcium phosphate (CaP scaffolds combined with bone morphogenetic proteins (BMPs or mesenchymal stem cells (MSCs in the field of bone tissue engineering (BTE. Date Sources: Data cited in this review were obtained primarily from PubMed and Medline in publications from 1979 to 2014, with highly regarded older publications also included. The terms BTE, CaP, BMPs, and MSC were used for the literature search. Study Selection: Reviews focused on relevant aspects and original articles reporting in vitro and/or in vivo results concerning the efficiency of CaP/BMPs or CaP/MSCs composites were retrieved, reviewed, analyzed, and summarized. Results: An ideal BTE product contains three elements: Scaffold, growth factors, and stem cells. CaP-based scaffolds are popular because of their outstanding biocompatibility, bioactivity, and osteoconductivity. However, they lack stiffness and osteoinductivity. To solve this problem, composite scaffolds of CaP with BMPs have been developed. New bone formation by CaP/BMP composites can reach levels similar to those of autografts. CaP scaffolds are compatible with MSCs and CaP/MSC composites exhibit excellent osteogenesis and stiffness. In addition, a CaP/MSC/BMP scaffold can repair bone defects more effectively than an autograft. Conclusions: Novel BTE products possess remarkable osteoconduction and osteoinduction capacities, and exhibit balanced degradation with osteogenesis. Further work should yield safe, viable, and efficient materials for the repair of bone lesions.

  17. Deer Bone Oil Extract Suppresses Lipopolysaccharide-Induced Inflammatory Responses in RAW264.7 Cells.

    Science.gov (United States)

    Choi, Hyeon-Son; Im, Suji; Park, Yooheon; Hong, Ki-Bae; Suh, Hyung Joo

    2016-01-01

    The aim of this study was to investigate the effect of deer bone oil extract (DBOE) on lipopolysaccharide (LPS)-induced inflammatory responses in RAW264.7 cells. DBOE was fractionated by liquid-liquid extraction to obtain two fractions: methanol fraction (DBO-M) and hexane fraction (DBO-H). TLC showed that DBO-M had relatively more hydrophilic lipid complexes, including unsaturated fatty acids, than DBOE and DBO-H. The relative compositions of tetradecenoyl carnitine, α-linoleic acid, and palmitoleic acid increased in the DBO-M fraction by 61, 38, and 32%, respectively, compared with DBOE. The concentration of sugar moieties was 3-fold higher in the DBO-M fraction than DBOE and DBO-H. DBO-M significantly decreased LPS-induced nitric oxide (NO) production in RAW264.7 cells in a dose-dependent manner. This DBO-M-mediated decrease in NO production was due to downregulation of mRNA and protein levels of inducible nitric oxide synthase (iNOS). In addition, mRNA expression of pro-inflammatory mediators, such as cyclooxygenase (COX-2), interleukin (IL)-1β, and IL-12β, was suppressed by DBO-M. Our data showed that DBO-M, which has relatively higher sugar content than DBOE and DBO-H, could play an important role in suppressing inflammatory responses by controlling pro-inflammatory cytokines and mediators. PMID:27040632

  18. Yellow Mealworm Protein for Food Purposes - Extraction and Functional Properties.

    Directory of Open Access Journals (Sweden)

    Xue Zhao

    Full Text Available A protocol for extraction of yellow mealworm larvae proteins was established, conditions were evaluated and the resulting protein extract was characterised. The freeze-dried yellow mealworm larvae contained around 33% fat, 51% crude protein and 43% true protein on a dry matter basis. The true protein content of the protein extract was about 75%, with an extraction rate of 70% under optimised extraction conditions using 0.25 M NaOH, a NaOH solution:ethanol defatted worm ratio of 15:1 mL/g, 40°C for 1 h and extraction twice. The protein extract was a good source of essential amino acids. The lowest protein solubility in distilled water solution was found between pH 4 and 5, and increased with either increasing or decreasing pH. Lower solubility was observed in 0.5 M NaCl solution compared with distilled water. The rheological tests indicated that temperature, sample concentration, addition of salt and enzyme, incubation time and pH alterations influenced the elastic modulus of yellow mealworm protein extract (YMPE. These results demonstrate that the functional properties of YMPE can be modified for different food applications.

  19. Yellow Mealworm Protein for Food Purposes - Extraction and Functional Properties.

    Science.gov (United States)

    Zhao, Xue; Vázquez-Gutiérrez, José Luis; Johansson, Daniel P; Landberg, Rikard; Langton, Maud

    2016-01-01

    A protocol for extraction of yellow mealworm larvae proteins was established, conditions were evaluated and the resulting protein extract was characterised. The freeze-dried yellow mealworm larvae contained around 33% fat, 51% crude protein and 43% true protein on a dry matter basis. The true protein content of the protein extract was about 75%, with an extraction rate of 70% under optimised extraction conditions using 0.25 M NaOH, a NaOH solution:ethanol defatted worm ratio of 15:1 mL/g, 40°C for 1 h and extraction twice. The protein extract was a good source of essential amino acids. The lowest protein solubility in distilled water solution was found between pH 4 and 5, and increased with either increasing or decreasing pH. Lower solubility was observed in 0.5 M NaCl solution compared with distilled water. The rheological tests indicated that temperature, sample concentration, addition of salt and enzyme, incubation time and pH alterations influenced the elastic modulus of yellow mealworm protein extract (YMPE). These results demonstrate that the functional properties of YMPE can be modified for different food applications. PMID:26840533

  20. Automatic segmentation of the bone and extraction of the bone-cartilage interface from magnetic resonance images of the knee

    International Nuclear Information System (INIS)

    The accurate segmentation of the articular cartilages from magnetic resonance (MR) images of the knee is important for clinical studies and drug trials into conditions like osteoarthritis. Currently, segmentations are obtained using time-consuming manual or semi-automatic algorithms which have high inter- and intra-observer variabilities. This paper presents an important step towards obtaining automatic and accurate segmentations of the cartilages, namely an approach to automatically segment the bones and extract the bone-cartilage interfaces (BCI) in the knee. The segmentation is performed using three-dimensional active shape models, which are initialized using an affine registration to an atlas. The BCI are then extracted using image information and prior knowledge about the likelihood of each point belonging to the interface. The accuracy and robustness of the approach was experimentally validated using an MR database of fat suppressed spoiled gradient recall images. The (femur, tibia, patella) bone segmentation had a median Dice similarity coefficient of (0.96, 0.96, 0.89) and an average point-to-surface error of 0.16 mm on the BCI. The extracted BCI had a median surface overlap of 0.94 with the real interface, demonstrating its usefulness for subsequent cartilage segmentation or quantitative analysis

  1. Aqueous extract of pomegranate seed attenuates glucocorticoid-induced bone loss and hypercalciuria in mice: A comparative study with alendronate.

    Science.gov (United States)

    Zhang, Yan; Shao, Jin; Wang, Zhi; Yang, Tieyi; Liu, Shuyi; Liu, Yue; Fan, Xinbing; Ye, Weiguang

    2016-08-01

    The present study was performed in order to examine bone loss and calcium homeostasis in mice with glucocorticoid (GC)-induced osteoporosis (GIOP) following treatment with the aqueous extract of pomegranate seed (AE-PS). In addition, a comparative study with alendronate was performed. Biomarkers in the serum and the urine were measured. The tibias, kidney and duodenum were removed in order to measure the levels of bone calcium, protein expression as well as to perform histomorphological analysis of the bone. GC treatment facilitated the induction of hypercalciuria in the mice, and the AE-PS‑treated mice exhibited a greater increase in serum calcium and a decrease in urine calcium. The AE-PS reversed the deleterious effects on the trabecular bone induced by DXM and stimulated bone remodeling, including an increase in bone calcium and alkaline phosphatase‑b (ALP-b) and a decrease in a the critical bone resorption markers C-terminal telopeptide of type I collagen (CTX) and tartrate‑resistant acid phosphatase-5b (TRAP-5b). Hematoxylin and eosin (H&E) staining revealed the increased disconnections and separation between the growth plate and the trabecular bone network as well as the reduction in the trabecular bone mass of the primary and secondary spongiosa throughout the proximal metaphysis of the tibia in the DXM group. Moreover, the decreased protein expression of transient receptor potential vanilloid (TRPV)5, TRPV6 and calbindin‑D9k (CaBP‑9k) was reversed by the AE-PS or alendronate supplementation in the kidneys and the duodenum as well as plasma membrane Ca2+‑ATPase1 (PMCA1) expression in the kidneys of mice with GIOP. There was no marked difference in pharmacological effectiveness between alendronate and the AE-PS. Taken together, these findings suggest that the AE-PS may be an alternative therapy suitable for use in the management of secondary osteoporosis. PMID:27278225

  2. AMP-activated protein kinase (AMPK) activation regulates in vitro bone formation and bone mass

    OpenAIRE

    Shah, M; Kola, B; Bataveljic, A.; Arnett, T. R.; Viollet, B.; Saxon, L.; Korbonits, M.; C. Chenu

    2010-01-01

    Adenosine 5′-monophosphate-activated protein kinase (AMPK), a regulator of energy homeostasis, has a central role in mediating the appetite-modulating and metabolic effects of many hormones and antidiabetic drugs metformin and glitazones. The objective of this study was to determine if AMPK can be activated in osteoblasts by known AMPK modulators and if AMPK activity is involved in osteoblast function in vitro and regulation of bone mass in vivo. ROS 17/2.8 rat osteoblast-like cells were cult...

  3. Gene gun transferring-bone morphogenetic protein 2 (BMP-2) gene enhanced bone fracture healing in rabbits

    OpenAIRE

    Li, Wenju; Wei, Haifeng; Xia, Chunmei; Zhu, Xiaomeng; Hou, Guozhu; Xu, Feng; Xinghua SONG; Zhan, Yulin

    2015-01-01

    Purpose: Transferring the bone morphogenetic protein 2 (BMP-2) genes into the tissues or cells can improve the bone healing of the fracture has been widely accepted. We evaluated the efficiency of using gene gun to transfer the BMP-2 gene thereby affected the healing of a fractured bone. Methods: The vector coding for BMP-2 was constructed by a non-replicating encephalo-myocarditis virus (ECMV)-based vector. The segmental bone defect (1.5 cm) model was created by a wire-saw at the middle part...

  4. Comparison of Methods for Protein Extraction from Pine Needles

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Extraction of proteins from pine needles for proteomic analysis has long been a challenge for scientists. We compared three different protein extraction methods including sucrose, Tris-HCl and trichloroacetic acid (TCA)/acetone (TCA method) to determine their efficiency in separating pine needle proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional PAGE (2D-PAGE). Proteins were then separated by SDS-PAGE. Among three methods the method using sucrose extraction buffer showed the highest efficiency and highest quality in separating proteins. In addition, clearer and more stable strips were detected by SDS-PAGE using sucrose extraction buffer. When the proteins extracted using sucrose extraction buffer were separated by 2D-PAGE, more than 300 protein spots, with isoelectric points (PI) ranging from 4.0 to 7.0 and molecular weights (MW) from 6.5 to 97.4 kD, were observed. This confirmed that the method with sucrose extraction buffer was an efficient and reliable method for extracting proteins from pine needles.

  5. Biochemical Characterization of Major Bone-Matrix Proteins Using Nanoscale-Size Bone Samples and Proteomics Methodology*

    OpenAIRE

    Grażyna E Sroga; Karim, Lamya; Colón, Wilfredo; Vashishth, Deepak

    2011-01-01

    There is growing evidence supporting the need for a broad scale investigation of the proteins and protein modifications in the organic matrix of bone and the use of these measures to predict fragility fractures. However, limitations in sample availability and high heterogeneity of bone tissue cause unique experimental and/or diagnostic problems. We addressed these by an innovative combination of laser capture microscopy with our newly developed liquid chromatography separation methods, follow...

  6. Preparation of protein samples for gel electrophoresis by sequential extraction

    Institute of Scientific and Technical Information of China (English)

    钟伯雄; 翁宏飚; 等

    2002-01-01

    Since preparation and solubilization of protein samples are crucial factors in proteome research,the authors established a sequential extraction technique to prepare protein samples from the body wall of the 5th instar larvae of silkworm.Bombyx mori.Two kinds of protein samples were obtained from the body wall using the method.Between the two types of samples only about 15% proteins were identical;the majority were different,indicating that more species of proteins could be obtained with the sequential extraction method;which will be useful for preparation of protein samples for proteome study.

  7. Extractable protein content of radiation vulcanized natural rubber latex films

    International Nuclear Information System (INIS)

    The effects of processing conditions on extractable protein content of coagulant dipped radiation vulcanized natural rubber latex films have been investigated. Drying of wet-gel of radiation vulcanized latex films even at a relatively low temperature of 70 degree C resulted in increases of extractable protein content of the films. The extractable protein content is dependent upon both the temperature and time of drying of wet-gel deposit. Wet-gel leaching of film alone is not adequate to reduce the extractable protein content of films to low levels. Combination of wet-gel leaching, post-leaching, a dip in corn starch slurry, followed by drying at a low temperature of 70 degree C reduces the extractable protein content of films to very low levels

  8. Bone graft substitutes and bone morphogenetic proteins for osteoporotic fractures: what is the evidence?

    Science.gov (United States)

    Van Lieshout, Esther M M; Alt, Volker

    2016-01-01

    Despite improvements in implants and surgical techniques, osteoporotic fractures remain challenging to treat. Among other major risk factors, decreased expression of morphogenetic proteins has been identified for impaired fracture healing in osteoporosis. Bone grafts or bone graft substitutes are often used for stabilizing the implant and for providing a scaffold for ingrowth of new bone. Both synthetic and naturally occurring biomaterials are available. Products generally contain hydroxyapatite, tricalcium phosphate, dicalcium phosphate, calcium phosphate cement, calcium sulfate (plaster of Paris), or combinations of the above. Products have been used for the treatment of osteoporotic fractures of the proximal humerus, distal radius, vertebra, hip, and tibia plateau. Although there is generally consensus that screw augmentation increased the biomechanical properties and implant stability, the results of using these products for void filling are not unequivocal. In osteoporotic patients, Bone Morphogenetic Proteins (BMPs) have the potential impact to improve fracture healing by augmenting the impaired molecular and cellular mechanisms. However, the clinical evidence on the use of BMPs in patients with osteoporotic fractures is poor as there are no published clinical trials, case series or case studies. Even pre-clinical literature on in vitro and in vivo data is weak as most articles focus on the beneficial role for BMPs for restoration of the underlying pathophysiological factors of osteoporosis but do not look at the specific effects on osteoporotic fracture healing. Limited data on animal experiments suggest stimulation of fracture healing in ovariectomized rats by the use of BMPs. In conclusion, there is only limited data on the clinical relevance and optimal indications for the use of bone graft substitute materials and BMPs on the treatment of osteoporotic fractures despite the clinical benefits of these materials in other clinical indications. Given the

  9. Up-regulation of bone marrow stromal protein 2 (BST2 in breast cancer with bone metastasis

    Directory of Open Access Journals (Sweden)

    Zheng Xin

    2009-04-01

    Full Text Available Abstract Background Bone metastases are frequent complications of breast cancer. Recent literature implicates multiple chemokines in the formation of bone metastases in breast cancer. However, the molecular mechanism of metastatic bone disease in breast cancer remains unknown. We have recently made the novel observation of the BST2 protein expression in human breast cancer cell lines. The purpose of our present study is to investigate the expression and the role of BST2 in bone metastatic breast cancer. Methods cDNA microarray analysis was used to compare the BST2 gene expression between a metastatic to bone human breast cancer cell line (MDA-231BO and a primary human breast cancer cell line (MDA-231. The BST2 expression in one bone metastatic breast cancer and seven non-bone metastatic breast cancer cell lines were also determined using real-time RT-PCR and Western blot assays. We then employed tissue array to further study the BST2 expression in human breast cancer using array slides containing 20 independent breast cancer tumors that formed metastatic bone lesions, 30 non-metastasis-forming breast cancer tumors, and 8 normal breast tissues. In order to test the feasibility of utilizing BST2 as a serum marker for the presence of bone metastasis in breast cancer, we had measured the BST2 expression levels in human serums by using ELISA on 43 breast cancer patients with bone metastasis, 43 breast cancer patients without bone metastasis, and 14 normal healthy controls. The relationship between cell migration and proliferation and BST2 expression was also studied in a human breast recombinant model system using migration and FACS analysis. Results The microarray demonstrated over expression of the BST2 gene in the bone metastatic breast cancer cell line (MDA-231BO compared to the primary human breast cancer cell line (MDA-231. The expression of the BST2 gene was significantly increased in the bone metastatic breast cancer cell lines and tumor

  10. Up-regulation of bone marrow stromal protein 2 (BST2) in breast cancer with bone metastasis

    International Nuclear Information System (INIS)

    Bone metastases are frequent complications of breast cancer. Recent literature implicates multiple chemokines in the formation of bone metastases in breast cancer. However, the molecular mechanism of metastatic bone disease in breast cancer remains unknown. We have recently made the novel observation of the BST2 protein expression in human breast cancer cell lines. The purpose of our present study is to investigate the expression and the role of BST2 in bone metastatic breast cancer. cDNA microarray analysis was used to compare the BST2 gene expression between a metastatic to bone human breast cancer cell line (MDA-231BO) and a primary human breast cancer cell line (MDA-231). The BST2 expression in one bone metastatic breast cancer and seven non-bone metastatic breast cancer cell lines were also determined using real-time RT-PCR and Western blot assays. We then employed tissue array to further study the BST2 expression in human breast cancer using array slides containing 20 independent breast cancer tumors that formed metastatic bone lesions, 30 non-metastasis-forming breast cancer tumors, and 8 normal breast tissues. In order to test the feasibility of utilizing BST2 as a serum marker for the presence of bone metastasis in breast cancer, we had measured the BST2 expression levels in human serums by using ELISA on 43 breast cancer patients with bone metastasis, 43 breast cancer patients without bone metastasis, and 14 normal healthy controls. The relationship between cell migration and proliferation and BST2 expression was also studied in a human breast recombinant model system using migration and FACS analysis. The microarray demonstrated over expression of the BST2 gene in the bone metastatic breast cancer cell line (MDA-231BO) compared to the primary human breast cancer cell line (MDA-231). The expression of the BST2 gene was significantly increased in the bone metastatic breast cancer cell lines and tumor tissues compared to non-bone metastatic breast cancer

  11. Extractability and characteristics of proteins deriving from wheat DDGS.

    Science.gov (United States)

    Chatzifragkou, Afroditi; Prabhakumari, Parvathy Chandran; Kosik, Ondrej; Lovegrove, Alison; Shewry, Peter R; Charalampopoulos, Dimitrios

    2016-05-01

    Wheat Distillers' Dried Grains with Solubles (DDGS) and in-process samples were used for protein extraction. Prolamins were the predominant protein components in the samples. The absence of extractable α- and γ-gliadins in DDGS indicated protein aggregation during the drum drying processing stage. Prolamin extraction was performed using 70% (v/v) ethanol or alkaline-ethanol solution in the presence of reducing agent. DDGS extracts had relatively low protein contents (14-44.9%, w/w), regardless of the condition applied. The wet solids were the most suitable raw material for protein extraction, with recovery yields of ∼ 55% (w/w) and protein content of ∼ 58% (w/w) in 70% (v/v) ethanol. Protein extracts from wet solids were significantly rich in glutamic acid and proline. Mass balance calculations demonstrated the high carbohydrate content (∼ 50%, w/w) of solid residues. Overall, the feasibility of utilising in-process samples of DDGS for protein extraction with commercial potential was demonstrated. PMID:26769499

  12. Effect of Green Tea Extract in Reducing Genotoxic Injuries of Cell Phone Microwaves on Bone Marrow

    OpenAIRE

    Zahra Zahedifar; Javad Baharara

    2013-01-01

    Background: Green tea (Camellia sinensis) extract is rich source of natural antioxidants specially catechin that is quickly absorbed into the body and it has cancer protective, anti microbial and anti inflammation effects. In this study has been studied role of green tea extract against genotoxic damage induced by cell phone microwaves on bone marrow polychromatic erythrocytes of adult male Balb/C mouse.Materials and Methods: In this experimental study 40 mouse were divided into five groups, ...

  13. Radioprotective effects of hawthorn fruit extract against gamma irradiation in mouse bone marrow cells

    International Nuclear Information System (INIS)

    The radioprotective effect of hawthorn (Crataegus microphylla) fruit extract against genotoxicity induced by gamma irradiation has been investigated in mouse bone marrow cells. A single intraperitoneal (ip) administration of hawthorn extract at doses of 25, 50, 100 and 200 mg/kg 1 h prior to gamma irradiation (2 Gy) reduced the frequencies of micronucleated polychromatic erythrocytes (MnPCEs). All four doses of hawthorn extract significantly reduced the frequencies of MnPCEs and increased the PCE/PCE+NCE ratio (polychromatic erythrocyte/polychromatic erythrocyte+normochromatic erythrocyte) in mice bone marrow compared with the non drug-treated irradiated control (p<0.02-0.00001). The maximum reduction in MnPCEs was observed in mice treated with extract at a dose of 200 mg/kg. Administration of amifostine at dose 100 mg/kg and hawthorn at dose 200 mg/kg reduced the frequency of MnPCE almost 4.8 and 5.7 fold; respectively, after being exposed to 2 Gy of gamma rays, compare with the irradiated control group. Crataegus extract exhibited concentration-dependent activity on 1, 1-diphenyl 2-picrylhydrazyl free radical showing that Crataegus contained high amounts of phenolic compounds and the high performance liquid chromatography (HPLC) analysis determined that it contained chlorogenic acid, epicatechin and hyperoside. It appeared that hawthorn extract with antioxidant activity reduced the genotoxicity induced by gamma irradiation in bone marrow cells. (author)

  14. A novel, truncated human bone morphogenetic protein-2:construction, expression ,functions and clinical potential

    Institute of Scientific and Technical Information of China (English)

    XU Fang

    2001-01-01

    @@ Introduction As a member of the bone morphogenetic protein (BMP) family, BMP-2 plays important roles not only in bone regeneration and bone repair but also in cell proliferation, apoptosis, differentiation and morphogenesis. The BMP-2 remarkable ability to stimulate new bone growth results in the development of a novel therapy strategy for bone mass defect due to accidents or diseases. Because the BMP-2 itself, in conjunction with a suitable matrix, is sufficient to stimulate genesis of new bone, the genetically engineered BMP-2 has good applied prospects.

  15. Mesenchymal stromal cells from bone marrow treated with bovine tendon extract acquire the phenotype of mature tenocytes

    OpenAIRE

    Lívia Maria Mendonça Augusto; Diego Pinheiro Aguiar; Danielle Cabral Bonfim; Amanda dos Santos Cavalcanti; Priscila Ladeira Casado; Maria Eugênia Leite Duarte

    2016-01-01

    ABSTRACT OBJECTIVE: This study evaluated in vitro differentiation of mesenchymal stromal cells isolated from bone marrow, in tenocytes after treatment with bovine tendon extract. METHODS: Bovine tendons were used for preparation of the extract and were stored at -80 °C. Mesenchymal stromal cells from the bone marrow of three donors were used for cytotoxicity tests by means of MTT and cell differentiation by means of qPCR. RESULTS: The data showed that mesenchymal stromal cells from bone...

  16. Extraction of bone structure with a single-scan skeletonization driven by distance

    OpenAIRE

    Arlicot, Aurore; NORMAND, Nicolas; Amouriq, Yves; Guédon, Jeanpierre

    2011-01-01

    Shape description is an important step in image analysis. Skeletonization methods are widely used in image analysis since they are a powerful tool to describe a shape. This paper presents a new single-scan skeletonization using different diskrete distances. The application of this method is the extraction of caracteristics from µCT images in order to estimate the bone state.

  17. Bone morphogenetic protein-2: a potential regulator in scleral remodeling

    Science.gov (United States)

    Hu, Jianmin; Cui, Dongmei; Yang, Xiao; Wang, Shaowei; Hu, Shoulong; Li, Chuanxu

    2008-01-01

    Purpose Bone morphogenetic protein 2 (BMP-2) is a member of the main subgroup of bone morphogenetic proteins within the transforming growth factor-β superfamily. BMP-2 is involved in numerous cellular functions including development, cell proliferation, apoptosis, and extracellular matrix synthesis. We examined BMP-2 expression in human scleral fibroblasts (HSF) and assessed the effects of recombinant human BMP-2 (rhBMP-2) on HSF proliferation, matrix metalloproteinase-2 (MMP-2), and tissue inhibitor of metalloproteinase-2 (TIMP-2). Methods We used confocal fluorescence microscopy (CFM) to study BMP-2 distribution in HSF cells and frozen human scleral sections. The influence of rhBMP-2 on cell proliferation at different concentrations (0 ng/ml, 1 ng/ml, 10 ng/ml, and 100 ng/ml) was evaluated by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. The effects of rhBMP-2 on the cell cycle were investigated with flow cytometric analysis. Reverse transcription polymerase chain reaction (RT–PCR) and enzyme-linked immunosorbent assay (ELISA) were used to examine MMP-2 and TIMP-2 mRNAs and secreted proteins in HSF that were incubated with rhBMP-2. Results BMP-2 protein expression from human sclera was confirmed by CFM. Cell proliferation was significantly increased with 100 ng/ml rhBMP-2 in a time-dependent manner (p<0.05). The HSF cell cycle moved to the S and S+G2M phases after rhBMP-2 stimulation at 100 ng/ml compared to normal cells (p<0.05). TIMP-2 mRNA levels were significantly increased in HSF incubated for 24 h with 100 ng/ml rhBMP-2 (p<0.01). A 48 h incubation with 10 ng/ml or 100 ng/ml rhBMP-2 resulted in significantly increased TIMP-2 mRNA and protein expression and significantly decreased MMP-2 mRNA expression (p<0.01) while MMP-2 protein expression significantly decreased at 100 ng/ml rhBMP-2 (p<0.01). Conclusions Human sclera fibroblasts expressed BMP-2, which promoted cell proliferation, and elicited changes in MMP-2 and TIMP-2

  18. A randomized controlled evaluation of alveolar ridge preservation following tooth extraction using deproteinized bovine bone mineral and demineralized freeze-dried bone allograft

    Directory of Open Access Journals (Sweden)

    Rokhsareh Sadeghi

    2016-01-01

    Conclusion: Based on the findings of this study, both materials have positive effect on alveolar ridge preservation after tooth extraction, but there was more new bone formation and less residual graft particles in DFDBA group than in DBBM group.

  19. Fish Muscle Proteins: Extraction, Quantitation, and Electrophoresis

    Science.gov (United States)

    Smith, Denise

    Electrophoresis can be used to separate and visualize proteins. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), proteins are separated based on size. When protein samples are applied to such gels, it is usually necessary to know the protein content of the sample. This makes it possible to apply a volume of sample to the gel such that samples have a comparable amount of total protein. While it is possible to use an official method of protein analysis (e.g., Kjeldahl, N combustion) for such an application, it often is convenient to use a rapid spectroscopic protein analysis that requires only a small amount of sample. The bicinchoninic acid (BCA) assay method will be used for this purpose.

  20. Effects of casein, whey and soy proteins on volumetric bone density and bone strength in immunocompromised piglets

    DEFF Research Database (Denmark)

    Budek, Alicja Zofia; Bjørnvad, Charlotte; Mølgaard, Christian;

    2007-01-01

    Summary:Background and aims: Bone-promoting effect of different proteins in early life, under immunocompromised conditions, is unknown. We investigated effects of milk- and plantderived proteins on bone development in immunocompromised piglets. Methods: Newborn, colostrum-deprived piglets were...... assigned to a formula based on either casein (n=11), whey (n=11) or soy (n=10) as the protein source (each 55 g/L), and equal amounts of fat, carbohydrates, calcium and phosphorus. Results & Conclusion: Despite efforts to sustain immuno-protection (sow serum and antibiotic injections), some piglets became...

  1. Extractable proteins from field radiation vulcanized natural rubber latex

    International Nuclear Information System (INIS)

    The type I allergy associated with the use of natural rubber latex (NRL) products is caused by the NRL proteins leached by the sweat or other body fluids. Makuuchi's group proposed for the first time the proteins removal by the addition of water-soluble polymers (WSP) on radiation vulcanization of natural rubber latex (RVNRL) that is a promising process under development in many countries. In this study, Brazilian field natural rubber was irradiated with a 60Co gamma source to reduce the content of WSP in the final product. WSP was used as additive to improve the extraction of protein. After irradiation the RVNRL was centrifuged to extract the WSP and proteins. The analytical methodology for protein content was based on the modified Lowry method according to ASTM D5712. Protein determination was carried out in serum of latex and in the extracts of the gloves. The concentration of extractable water-soluble proteins in serum of irradiated field NRL (NRL1), not irradiated one (NRL2); of twice centrifuged sample with polymer additive NRL (NRL3) and of the glove manufactured (NRLG) are compared with commercial glove (CG). The irradiation process increases the extractable water-soluble proteins, EP, as reported in the literature. In this study the use of polymeric additive on the bi-centrifugation process to remove protein was successful and the EP of the glove obtained in NRL3 was at around 40% of the commercial glove

  2. Microdialysis Sampling from Wound Fluids Enables Quantitative Assessment of Cytokines, Proteins, and Metabolites Reveals Bone Defect-Specific Molecular Profiles

    Science.gov (United States)

    Wissenbach, Dirk K.; Pfeiffer, Susanne E. M.; Baumann, Sven; Hofbauer, Lorenz C.; von Bergen, Martin; Kalkhof, Stefan; Rammelt, Stefan

    2016-01-01

    Bone healing involves a variety of different cell types and biological processes. Although certain key molecules have been identified, the molecular interactions of the healing progress are not completely understood. Moreover, a clinical routine for predicting the quality of bone healing after a fracture in an early phase is missing. This is mainly due to a lack of techniques to comprehensively screen for cytokines, growth factors and metabolites at their local site of action. Since all soluble molecules of interest are present in the fracture hematoma, its in-depth assessment could reveal potential markers for the monitoring of bone healing. Here, we describe an approach for sampling and quantification of cytokines and metabolites by using microdialysis, combined with solid phase extractions of proteins from wound fluids. By using a control group with an isolated soft tissue wound, we could reveal several bone defect-specific molecular features. In bone defect dialysates the neutrophil chemoattractants CXCL1, CXCL2 and CXCL3 were quantified with either a higher or earlier response compared to dialysate from soft tissue wound. Moreover, by analyzing downstream adaptions of the cells on protein level and focusing on early immune response, several proteins involved in the immune cell migration and activity could be identified to be specific for the bone defect group, e.g. immune modulators, proteases and their corresponding inhibitors. Additionally, the metabolite screening revealed different profiles between the bone defect group and the control group. In summary, we identified potential biomarkers to indicate imbalanced healing progress on all levels of analysis. PMID:27441377

  3. Regulation of Bone Morphogenetic Protein Signaling by ADP-ribosylation.

    Science.gov (United States)

    Watanabe, Yukihide; Papoutsoglou, Panagiotis; Maturi, Varun; Tsubakihara, Yutaro; Hottiger, Michael O; Heldin, Carl-Henrik; Moustakas, Aristidis

    2016-06-10

    We previously established a mechanism of negative regulation of transforming growth factor β signaling mediated by the nuclear ADP-ribosylating enzyme poly-(ADP-ribose) polymerase 1 (PARP1) and the deribosylating enzyme poly-(ADP-ribose) glycohydrolase (PARG), which dynamically regulate ADP-ribosylation of Smad3 and Smad4, two central signaling proteins of the pathway. Here we demonstrate that the bone morphogenetic protein (BMP) pathway can also be regulated by the opposing actions of PARP1 and PARG. PARG positively contributes to BMP signaling and forms physical complexes with Smad5 and Smad4. The positive role PARG plays during BMP signaling can be neutralized by PARP1, as demonstrated by experiments where PARG and PARP1 are simultaneously silenced. In contrast to PARG, ectopic expression of PARP1 suppresses BMP signaling, whereas silencing of endogenous PARP1 enhances signaling and BMP-induced differentiation. The two major Smad proteins of the BMP pathway, Smad1 and Smad5, interact with PARP1 and can be ADP-ribosylated in vitro, whereas PARG causes deribosylation. The overall outcome of this mode of regulation of BMP signal transduction provides a fine-tuning mechanism based on the two major enzymes that control cellular ADP-ribosylation. PMID:27129221

  4. Impact of Black seed (Nigella sativa extract on bone turnover markers in postmenopausal women with osteoporosis

    Directory of Open Access Journals (Sweden)

    N Valizadeh

    2010-01-01

    Full Text Available "n "n  "n  "nBackground and the purpose of the study: "nExperimental studies have shown that Ns (Nigella sativa seeds oil can increase bone formation and may have anabolic effects on bone loss. This study was conducted to investigate the beneficial impacts of the oil of Black seeds on bone turnover in osteoporotic postmenopausal women. "nMaterials and methods: A placebo controlled pilot study was carried out on 15 postmenopausal osteoporotic women of 48-74 years old. In addition to Calcium-D supplements (2 tablets per day all participants were randomly received Ns extract (3ml, 0.05 ml/kg/day p .o. or placebo for 3 months. In all subjects hematological tests were performed and hepatic enzymes, BUN, Cr, Ca, P and plasma bone formation and resorption markers including osteocalcin, bone alkaline phosphatase (Bone-ALP and carboxy terminal cross linked telopeptide (CTX was determined before and after 12 weeks of treatment. "nResults: Twelve participants completed the entire 12 weeks study course of which 5 and 7 women were belonged to Ns and placebo groups respectively. Women in placebo group were significantly older than women in Ns group. There were not significant differences between BMIs, BMD results and plasma levels of bone marker in two groups at the baseline and plasma levels of bone markers between Ns and placebo group at the end of 12 weeks. Alterations from baseline in bone markers levels did not differ significantly between two groups. We did not observe any side effects due to Ns therapy. "nConclusion: In this pilot study similar to the previous trial, we failed to show beneficial impact of Ns extract administration for a short time on bone turnover so we don’t suggest it for medicinal application in the osteoporosis condition. Long time duration studies with larger sample size and usage of a more tolerable dosage forms of Black seeds oil should be emphasized for further clarification of its useful anabolic effects on bone metabolism.

  5. Research report Organofinery protein extraction AAU

    OpenAIRE

    Molinuevo-Salces, Beatriz; Fernandez, Maria Santamaria; Lübeck,Mette

    2015-01-01

    The supply of organic protein feed for monogastric animals (i.e. poultry and pigs) with the right amino acid profile and a competitive price is one of the major challenges for organic agriculture nowadays. In this project, the development of an organic protein concentrate able to substitute the soybean protein while competing with the current market prices is assessed. The product is developed from red clover biomass, using a sustainable technology without the use of inorganic acids or organi...

  6. Preimplant preparation of the extraction alveolus with the deproteinized bovine bone and calcium-sulphate

    Directory of Open Access Journals (Sweden)

    Brković Božidar

    2006-01-01

    Full Text Available Background. Different materials are used to prevent the resorption of alveolar bone. The aim of this report was to show the radiographical and histological results prior to implant insertion, when a deproteinized bovine bone mineral (BioOss and calcium-sulphate were placed into the extraction socket immediately after the tooth removal. Case report. A 22-year-old woman was scheduled for the removal of the second lower molar when the extraction socket was filled with BioOss covered with calcium-sulphate as a resorbable membrane. Primary closure of the surgical site was performed. Radiography was done 4 and 12 months later. One year after the surgery, when the implant was inserted, a biopsy of the new regenerated bone was obtained for the histological evaluation. The lamellar bone was evident using both materials. The resorption of BioOss was slow and the connective tissue was observed. Conclusion. Both materials had biocompatible and oseoconductive properties. One year after the grafting procedure, we observed the lamellar bone and partial resorption of BioOss, while calciumsulphate showed no significant effect as a resorbable membrane.

  7. Optimal extraction and hydrolysis of Chlorella pyrenoidosa proteins.

    Science.gov (United States)

    Wang, Xiaoqin; Zhang, Xuewu

    2012-12-01

    In this study, for the first time, the applications of two new methods, ionic liquid and low-temperature high-pressure cell breakage methods, to the extraction of whole proteins in Chlorella pyrenoidosa cells were explored. Meanwhile, the comparison with three traditional methods was also made. The results indicated that the extraction rate for ionic liquid is only at moderate level, but the new low-temperature high-pressure cell breakage method can obviously increase the protein extraction rate up to 2- to 15-fold. Subsequently, the hydrolysis of the extracted proteins was conducted with three enzymes (papain, trypsin and alcalase). The data presented that the degree of hydrolysis for each enzyme under the optimal conditions is in the order of: alcalase (18.31%)>papain (14.33%)>trypsin (8.47%), demonstrating the potential of C. pyrenoidosa protein hydrolysates obtained here in nutritional supplement and medical foods. PMID:23117187

  8. Possible Role of Garlic Oil and Parsley Extract in Ameliorating Radiation-Induced Bone Loss in Female Rats

    International Nuclear Information System (INIS)

    To Investigate the possible protective effect of garlic oil and parsley extract against bone loss resulted in female virgin rats exposed to fractionated doses of gamma-radiation (1 Gy 3 times weekly for 5 weeks). Urinary calcium (U Ca), calcium to creatinine ratio (Ca/Cr), hydroxyproline and serum phosphorus were measured as bone resorption bio markers, while serum osteocalcine (OST) and serum alkaline phosphatase (ALP) were measured as bone formation bio markers. Furthermore, nitric oxide (NO) which represents the balance in bone remodeling was measured. Malondiadehyde level (MDA) as well as superoxide dismutase activity (SOD) was measured as oxidative stress bio markers. Female irradiated rats in the present study had significant increases in both bone resorption and bone formation bio markers after 6 weeks from the last exposure to gamma-radiation. Irradiated rats also had significant decreases in plasma NO indicating imbalance in bone remodeling as well as significant increase in oxidative stress bio markers. Daily treatment with garlic oil extracted in olive oil improved all measured parameters except OST level, while the vehicle used for garlic oil (extra virgin olive oil) significantly decreased bone resorption bio markers. Parsley extract induced normalization to all bone resorption and formation parameters measured in irradiated rats. Daily administration of garlic oil and parsley extract protected the bone from degeneration induced by exposure to fractionated doses of gamma radiation.

  9. Techno-economical evaluation of protein extraction for microalgae biorefinery

    Science.gov (United States)

    Sari, Y. W.; Sanders, J. P. M.; Bruins, M. E.

    2016-01-01

    Due to scarcity of fossil feedstocks, there is an increasing demand for biobased fuels. Microalgae are considered as promising biobased feedstocks. However, microalgae based fuels are not yet produced at large scale at present. Applying biorefinery, not only for oil, but also for other components, such as carbohydrates and protein, may lead to the sustainable and economical microalgae-based fuels. This paper discusses two relatively mild conditions for microalgal protein extraction, based on alkali and enzymes. Green microalgae (Chlorella fusca) with and without prior lipid removal were used as feedstocks. Under mild conditions, more protein could be extracted using proteases, with the highest yields for microalgae meal (without lipids). The data on protein extraction yields were used to calculate the costs for producing 1 ton of microalgal protein. The processing cost for the alkaline method was € 2448 /ton protein. Enzymatic method performed better from an economic point of view with € 1367 /ton protein on processing costs. However, this is still far from industrially feasible. For both extraction methods, biomass cost per ton of produced product were high. A higher protein extraction yield can partially solve this problem, lowering processing cost to €620 and 1180 /ton protein product, using alkali and enzyme, respectively. Although alkaline method has lower processing cost, optimization appears to be better achievable using enzymes. If the enzymatic method can be optimized by lowering the amount of alkali added, leading to processing cost of € 633/ton protein product. Higher revenue can be generated when the residue after protein extraction can be sold as fuel, or better as a highly digestible feed for cattle.

  10. Extracting knowledge from protein structure geometry

    DEFF Research Database (Denmark)

    Røgen, Peter; Koehl, Patrice

    2013-01-01

    Protein structure prediction techniques proceed in two steps, namely the generation of many structural models for the protein of interest, followed by an evaluation of all these models to identify those that are native-like. In theory, the second step is easy, as native structures correspond to m...

  11. Effects of injured spinal cord extracts on brain-derived neurotropic factor and myelin proteolipid protein in bone marrow mesenchymal stem cells%损伤脊髓匀浆上清对骨髓间充质干细胞分泌髓鞘前脂蛋白、脑源性神经营养因子的影响

    Institute of Scientific and Technical Information of China (English)

    刘然; 范东艳; 金鹏; 范洪学; 王苹

    2011-01-01

    背景:损伤脊髓匀浆上清成分复杂,其中不仅存在多种化学物质,而且也存在着多种细胞因子,这些物质能否影响骨髓间充质干细胞的增殖分化和分泌功能还不清楚.目的:探讨损伤脊髓匀浆上清成分对骨髓间充质干细胞分泌的脑源性神经营养因子和髓鞘前脂蛋白的影响.方法:贴壁法分离纯化Wistar大鼠骨髓间充质干细胞,稳定传到第3代后,分别用正常和损伤的Wistar大鼠脊髓匀浆上清诱导培养20 d.免疫荧光染色检测神经元特异性烯醇化酶阳性细胞,ELISA法检测培养液内髓鞘前脂蛋白、脑源性神经营养因子的含量,即时定量-PCR检测髓鞘前脂蛋白mRNA、脑源性神经营养因子mRNA水平.结果与结论:损伤脊髓匀浆上清液诱导培养骨髓间充质干细胞后,神经元特异性烯醇化酶阳性细胞率和培养液内脑源性神经营养因子、髓鞘前脂蛋白的含量在各个时间点均较正常脊髓匀浆上清液对骨髓间充质干细胞培养组高.提示,损伤的脊髓匀浆上清液能够诱导骨髓间充质干细胞分泌脑源性神经营养因子、髓鞘前脂蛋白,有利于向神经细胞方向分化.%BACKGROUND: The component of injured spinal cord extracts is complex and includes some chemical substances or cytokines.It remains unclear if these factors may influence cellular proliferation and secretion of bone marrow mesenchymal stem cells (BMSCs) or not.OBJECTIVE: To investigate the effect of injured spinal cord extracts on secretion of brain-derived neurotropic factor (BDNF)or myelin proteolipid protein (PLP) in BMSCs.METHODS: BMSCs were obtained from Wistar rats. After 3 generation of cell proliferation, BMSCs were induced with normal or injured spinal cord extracts for 20 days. Neuron specific enolase (NSE) positive cells were detected by immunofluorescence staining. Levels of BDNF and PLP in BMSCs cultured supernatant were evaluated using enzyme linked immunosorbent assay

  12. Deer Bone Extract Prevents Against Scopolamine-Induced Memory Impairment in Mice

    OpenAIRE

    Du, Chun Nan; Min, A Young; Kim, Hyun Jeong; Shin, Suk Kyung; Yu, Ha Ni; Sohn, Eun Jeong; Ahn, Chang-Won; Jung, Sung Ug; Park, Soo-Hyun; Kim, Mee Ree

    2015-01-01

    Deer bone has been used as a health-enhancing food as well as an antiaging agent in traditional Oriental medicine. Recently, the water extract of deer bone (DBE) showed a neuroprotective action against glutamate or Aβ1–42-induced cell death of mouse hippocampal cells by exerting antioxidant activity through the suppression of MAP kinases. The present study is to examine whether DBE improves memory impairment induced by scopolamine. DBE (50, 100 or 200 mg/kg) was administered orally to mice fo...

  13. Extracting accurate strain measurements in bone mechanics: A critical review of current methods.

    Science.gov (United States)

    Grassi, Lorenzo; Isaksson, Hanna

    2015-10-01

    Osteoporosis related fractures are a social burden that advocates for more accurate fracture prediction methods. Mechanistic methods, e.g. finite element models, have been proposed as a tool to better predict bone mechanical behaviour and strength. However, there is little consensus about the optimal constitutive law to describe bone as a material. Extracting reliable and relevant strain data from experimental tests is of fundamental importance to better understand bone mechanical properties, and to validate numerical models. Several techniques have been used to measure strain in experimental mechanics, with substantial differences in terms of accuracy, precision, time- and length-scale. Each technique presents upsides and downsides that must be carefully evaluated when designing the experiment. Moreover, additional complexities are often encountered when applying such strain measurement techniques to bone, due to its complex composite structure. This review of literature examined the four most commonly adopted methods for strain measurements (strain gauges, fibre Bragg grating sensors, digital image correlation, and digital volume correlation), with a focus on studies with bone as a substrate material, at the organ and tissue level. For each of them the working principles, a summary of the main applications to bone mechanics at the organ- and tissue-level, and a list of pros and cons are provided. PMID:26099201

  14. Honey protein extraction and determination by mass spectrometry.

    Science.gov (United States)

    Chua, Lee Suan; Lee, Jun You; Chan, Giek Far

    2013-04-01

    There are relatively limited studies on the protein of honey samples mainly because of the low amount of protein in honey (0.1-0.5 %), the difficulty in extracting honey protein from the sugar-rich environment, and the hindrance of protein characterization by conventional approaches. Several protein extraction methods such as mechanical (ultrafiltration and ultracentrifugation) and chemical (precipitation) techniques have been applied to different types of honey samples. Most of these studies reported the quantity and molecular size of honey protein from gel electrophoresis, but were unable to identify and characterize the protein. This limitation might be due to the low capacity of analytical equipment in those days. Although different precipitants have also been used, not all them are compatible with mass spectrometric methods during downstream analysis. As a result, the sample preparation step is essential in order to confidently characterize the low and varied amount of honey protein. Nowadays, honey protein is getting attention from researchers because of its potential activity in pharmacological applications. Therefore, honey protein extraction and determination by mass spectrometry are critically reviewed in order to stimulate further honey protein research. PMID:23292042

  15. Discontinuous release of bone morphogenetic protein-2 loaded within interconnected pores of honeycomb-like polycaprolactone scaffold promotes bone healing in a large bone defect of rabbit ulna.

    Science.gov (United States)

    Bae, Ji-Hoon; Song, Hae-Ryong; Kim, Hak-Jun; Lim, Hong-Chul; Park, Jung-Ho; Liu, Yuchun; Teoh, Swee-Hin

    2011-10-01

    The choice of an appropriate carrier and its microarchitectural design is integral in directing bone ingrowth into the defect site and determining its subsequent rate of bone formation and remodeling. We have selected a three-dimensional polycaprolactone (PCL) scaffold with an interconnected honeycomb-like porous structure to provide a conduit for vasculature ingrowth as well as an osteoconductive pathway to guide recruited cells responding to a unique triphasic release of osteoinductive bone morphogenetic proteins (BMP) from these PCL scaffolds. We hypothesize that the use of recombinant human bone morphogenetic protein 2 (rhBMP2)-PCL constructs promotes rapid union and bone regeneration of a large defect. Results of our pilot study on a unilateral 15 mm mid-diaphyseal segmental rabbit ulna defect demonstrated enhanced bone healing with greater amount of bone formation and bridging under plain radiography and microcomputed tomography imaging when compared with an empty PCL and untreated group after 8 weeks postimplantation. Quantitative measurements showed significantly higher bone volume fraction and trabecular thickness, with lower trabecular separation in the rhBMP2-treated groups. Histology evaluation also revealed greater mature bone formation spanning across the entire scaffold region compared with other groups, which showed no bone regeneration within the central defect zone. We highlight that it is the uniqueness of the scaffold having a highly porous network of channels that promoted vascular integration and allowed for cellular infiltration, leading to a discontinuous triphasic BMP2 release profile that mimicked the release profile during natural repair mechanisms in vivo. This study serves as preclinical evidence demonstrating the potential of combining osteoinductive rhBMP2 with our PCL constructs for the repair of large defects in a large animal model. PMID:21682591

  16. Radioprotective effects of citrus extract against γ-irradiation in mouse bone marrow cells

    International Nuclear Information System (INIS)

    The radioprotective effects of citrus extract were investigated by using the micronucleus test for anti-clastogenic and cell proliferation activity. A single intraperitoneal (i.p.) injection of citrus extract (Citrus aurantium var. amara) at 250, 500, 1,000 mg/kg body weight 1 h prior to γ-ray irradiation (1.5 Gy) reduced the frequencies of micronucleated polychromatic erythrocytes (MnPCEs) and normochromatic erythrocytes (MnNCEs). All three doses of citrus extract significantly reduced the frequencies of MnPCEs and MnNCEs in mice bone marrow compared to non-drug-treated irradiated control (p<0.005-0.05). The optimum dose for protection in mouse was 250 mg/kg to protect mice bone marrow 2.2-fold against the side effects of γ-irradiation with respect to the non-drug-treated irradiated control. The flavonoids were contained in citrus extract, probably to show protective activity, and reduced the clastogenic effect of radiation on mice bone marrow. Therefore fruits and vegetables contain flavonoids to be useful as protective effects under such stress conditions as irradiation. (author)

  17. Effect of Green Tea Extract in Reducing Genotoxic Injuries of Cell Phone Microwaves on Bone Marrow

    Directory of Open Access Journals (Sweden)

    Zahra Zahedifar

    2013-11-01

    Full Text Available Background: Green tea (Camellia sinensis extract is rich source of natural antioxidants specially catechin that is quickly absorbed into the body and it has cancer protective, anti microbial and anti inflammation effects. In this study has been studied role of green tea extract against genotoxic damage induced by cell phone microwaves on bone marrow polychromatic erythrocytes of adult male Balb/C mouse.Materials and Methods: In this experimental study 40 mouse were divided into five groups, control animals were located under natural condition, sham -exposed animals were prepared by experimental condition without cell phone waves radiation. Experimental 1 group that irradiated at cell phones for 4 days (3 hours/day and experimental 2 groups were injected intraperitoneal 100 mg/kg green tea extract for 5 days and experimental 3 group that irradiated at active mobile phones for 4 days (3 hours/day and were injected intraperitoneal 100 mg/kg green tea extract for 5 days. After treatment period micronucleus test was evaluated in polychromatic erythrocytes on bone marrow. The quantitative data was analyzed by ANOVA and Tukey test with using of SPSS-13 software at the level of p<0.05.Results: Based on this study, treatment with extracts of green tea decreased micronucleus frequency in bone marrow polychromatic erythrocytes of Balb/C mouse that irradiated at cell phone microwave (0.92±0.129, (p<0.001.Conclusion: Cell phone microwaves (940 MHz increased micronucleus on bone marrow polychromatic erythrocytes of male Balb/C mouse, but green tea had inhibitory effect and it decreased the average number of micronucleus.

  18. Ectopic bone formation of human bone morphogenetic protein-2 gene transfected goat bone marrow-derived mesenchymal stem cells in nude mice

    Institute of Scientific and Technical Information of China (English)

    汤亭亭; 徐小良; 戴尅戎; 郁朝锋; 岳冰; 楼觉人

    2005-01-01

    Objective: To evaluate the osteogenic potential of bone morphogenetic protein (BMP)-2 gene transfected goat bone marrow-derived mesenchymal stem cells (MSCs). Methods: Goat bone marrow- derived MSCs were transfected by Adv-human bone morphogenetic protein (hBMP)-2 gene(Group 1), Adv-beta gal transfected MSCs (Group 2)and uninfected MSCs(Group 3). Western blot analysis, alkaline phosphatase staining, Von Kossa staining and transmission electron microscopy were adopted to determine the phenotype of MSCs. Then the cells were injected into thigh muscles of the nude mice. Radiographical and histological evaluations were performed at different intervals. Results: Only Adv-hBMP-2 transfected MSCs produced hBMP-2. These cells were positive for alkaline phosphatase staining at the 12th day and were positive for Von Kossa staining at the 16th day after gene transfer. Electron microscopic observation showed that there were more rough endoplasmic reticulum, mitochondria and lysosomes in Adv-hBMP-2 transfected MSCs compared to MSCs of other two groups. At the 3rd and 6th weeks after cell injection, ectopic bones were observed in muscles of nude mice of Group 1. Only fibrous tissue or a little bone was found in other two groups. Conclusions: BMP-2 gene transfected MSCs can differentiate into osteoblasts in vitro and induce bone formation in vivo.

  19. Preventive effects of bee pollen Cistus ladaniferus extract on bone loss in ovariectomized rats in vivo

    International Nuclear Information System (INIS)

    The effect of bee pollen Cistus ladaniferus extract on ovariectomy (OVX)-induced bone loss in vivo was investigated. The water-solubilized extracts were obtained from the bee pollen of Cistus ladaniferus. Cistus extract (5.0 or 10.0 mg/100 g body weight) was orally administered once daily for 30 days to OVX rats. The analysis using a peripheral quantitative computed tomography (pQCT) showed that OVX-induced a significant decrease in mineral content, mineral density, and polar strength strain index in the femoral-metaphyseal tissues. These decreases were significantly prevented after the administration of Cistus extract (10.0 mg/100 g). Moreover, OVX-induced a significant decrease in calcium content in the femoral-diaphyseal and -metaphyseal tissues. This decrease was significantly prevented after the administration of cistus extract (5.0 or 10.0 mg/100 g). This study demonstrates that cistus extract has a preventive effect on OVX-induced bone loss in vivo. (author)

  20. Simvastatin enhances bone morphogenetic protein receptor type II expression

    International Nuclear Information System (INIS)

    Statins confer therapeutic benefits in systemic and pulmonary vascular diseases. Bone morphogenetic protein (BMP) receptors serve essential signaling functions in cardiovascular development and skeletal morphogenesis. Mutations in BMP receptor type II (BMPR2) are associated with human familial and idiopathic pulmonary arterial hypertension, and pathologic neointimal proliferation of vascular endothelial and smooth muscle cells within small pulmonary arteries. In severe experimental pulmonary hypertension, simvastatin reversed disease and conferred a 100% survival advantage. Here, modulation of BMPR2 gene expression by simvastatin is characterized in human embryonic kidney (HEK) 293T, pulmonary artery smooth muscle, and lung microvascular endothelial cells (HLMVECs). A 1.4 kb BMPR2 promoter containing Egr-1 binding sites confers reporter gene activation in 293T cells which is partially inhibited by simvastatin. Simvastatin enhances steady-state BMPR2 mRNA and protein expression in HLMVEC, through posttranscriptional mRNA stabilization. Simvastatin induction of BMPR2 expression may improve BMP-BMPR2 signaling thereby enhancing endothelial differentiation and function

  1. Evaluation of guided bone generation around implants placed into fresh extraction sockets: an experimental study in dogs

    DEFF Research Database (Denmark)

    Gotfredsen, K; Nimb, L; Buser, D; Hjørting-Hansen, E

    1993-01-01

    an implant is placed immediately into an extraction socket, it may not engage the walls of the socket near the crest of the alveolar ridge. With the presence of a bone defect around an implant, ingrowth of soft tissue could compromise the achievement of osseointegration in the crestal bone area. The......Immediate placement of implants into fresh extraction sockets would have the principal advantage of decreasing the recommended period of healing. It also would result in a guided placement of the implant, and it could reduce the resorption of the alveolar bone in the extraction area. However, when...... objective of this study was to evaluate the crestal bone healing response adjacent to implants placed immediately into fresh extraction sockets with and without covering membranes. Eight adult mongrel dogs had the third and fourth mandibular premolars extracted bilaterally. Thirty-two submerged titanium...

  2. Enzyme assisted protein extraction from rapeseed, soybean, and microalgae meals

    NARCIS (Netherlands)

    Sari, Y.W.; Bruins, M.E.; Sanders, J.P.M.

    2013-01-01

    Oilseed meals that are by-products from oil production are potential resources for protein. The aim of this work is to investigate the use of enzymes in assisting in the extraction of protein from different oilseed meals, namely rapeseed, soybean, and microalgae meals. In addition, microalgae withou

  3. Extractable proteins from irradiated field natural rubber latex

    International Nuclear Information System (INIS)

    Complete text of publication follows. Natural rubber latex (NRL) is extracted from Hevea brasiliensis tree and is used to produce different kinds of rubber goods like gloves, condoms, balloons and some part of medical and dental equipments, but it is predominantly used in the production of surgical and examination gloves. The allergy caused by latex products has become a serious problem; the sweat can remove proteins and allow contact with skin causing sensitization or allergic reactions. Some methods have been suggested to reduce the extractable protein content in latex gloves, as using low protein latex; proper leaching during the process of production etc. In this study field natural rubber latex was irradiated in different doses in a 60Co to reduce the water-soluble protein content in the final product. The protein content of the films obtained by casting method was extracted with phosphate buffer solution, pH 7 and was measured using Micro BCA Protein Assay kit. Also was measured protein in the serum samples of field NRL. The concentration of extractable proteins increased with increasing radiation dose

  4. A Biphasic Calcium Sulphate/Hydroxyapatite Carrier Containing Bone Morphogenic Protein-2 and Zoledronic Acid Generates Bone

    Science.gov (United States)

    Raina, Deepak Bushan; Isaksson, Hanna; Hettwer, Werner; Kumar, Ashok; Lidgren, Lars; Tägil, Magnus

    2016-01-01

    In orthopedic surgery, large amount of diseased or injured bone routinely needs to be replaced. Autografts are mainly used but their availability is limited. Commercially available bone substitutes allow bone ingrowth but lack the capacity to induce bone formation. Thus, off-the-shelf osteoinductive bone substitutes that can replace bone grafts are required. We tested the carrier properties of a biphasic, calcium sulphate and hydroxyapatite ceramic material, containing a combination of recombinant human bone morphogenic protein-2 (rhBMP-2) to induce bone, and zoledronic acid (ZA) to delay early resorption. In-vitro, the biphasic material released 90% of rhBMP-2 and 10% of ZA in the first week. No major changes were found in the surface structure using scanning electron microscopy (SEM) or in the mechanical properties after adding rhBMP-2 or ZA. In-vivo bone formation was studied in an abdominal muscle pouch model in rats (n = 6/group). The mineralized volume was significantly higher when the biphasic material was combined with both rhBMP-2 and ZA (21.4 ± 5.5 mm3) as compared to rhBMP-2 alone (10.9 ± 2.1 mm3) when analyzed using micro computed tomography (μ-CT) (p < 0.01). In the clinical setting, the biphasic material combined with both rhBMP-2 and ZA can potentially regenerate large volumes of bone. PMID:27189411

  5. Bone Tissue Engineering Using High Permeability Poly-epsilon-caprolactone Scaffolds Conjugated with Bone Morphogenetic Protein-2

    Science.gov (United States)

    Mitsak, Anna Guyer

    Bone is the second most commonly transplanted tissue in the United States. Limitations of current bone defect treatment options include morbidity at the autograft harvest site, mechanical failure, and poorly controlled growth factor delivery. Combining synthetic scaffolds with biologics may address these issues and reduce dependency on autografts. The ideal scaffolding system should promote tissue in-growth and nutrient diffusion, control delivery of biologics and maintain mechanical integrity during bone formation. This dissertation evaluates how scaffold permeability, conjugated bone morphogenetic protein-2 (BMP-2) and differentiation medium affect osteogenesis in vitro and bone growth in vivo.. "High" and "low" permeability polycaprolactone (PCL) scaffolds with regular architectures were manufactured using solid free form fabrication. Bone growth in vivo was evaluated in an ectopic mouse model. High permeability scaffolds promoted better 8 week bone growth, supported tissue penetration into the scaffold core, and demonstrated increased mechanical properties due to newly formed bone. Next, the effects of differentiation medium and conjugated BMP-2 on osteogenesis were compared. Conjugation may improve BMP-2 loading efficiency, help localize bone growth and control release. High permeability scaffolds were conjugated with BMP-2 using the crosslinker, sulfo-SMCC. When adipose-derived and bone marrow stromal cells were seeded onto constructs (with or without BMP-2), BMSC expressed more differentiation markers, and differentiation medium affected differentiation more than BMP-2. In vivo, scaffolds with ADSC pre-differentiated in osteogenic medium (with and without BMP-2) and scaffolds with only BMP-2 grew the most bone. Bone volume did not differ among these groups, but constructs with ADSC had evenly distributed, scaffold-guided bone growth. Analysis of two additional BMP-2 attachment methods (heparin and adsorption) showed highest conjugation efficiency for the

  6. Effects of bone morphogenetic protein-2 on bone cells in primary culture: immunohistochemical and electronmicroscopical studies

    Energy Technology Data Exchange (ETDEWEB)

    Schmitz, I.; Prochnow, N.; Mueller, K.M. [Berufsgenossenschaftliche Kliniken Bergmannsheil, Bochum (Germany). Inst. fuer Pathologie; Wiemann, M.; Schirrmacher, K.; Bingmann, D. [Essen Univ. (Germany). Inst. fuer Physiologie; Sebald, W. [Wuerzburg Univ. (Germany). Inst. fuer Physiologische Chemie II

    2001-02-01

    Bone morphogenetic protein 2 (BMP-2), among other morphogenetic effects on non osseous tissues, promotes bone formation in vivo. Therefore, BMP-2 may accelerate the integration of osseous implants. Although the effects of BMPs on cell proliferation have been studied extensively in vivo or in cell lines, little is published about effects on bone cells in primary cultures, especially on cell differentiation. As such information is a prerequisite to understand and to control effects of BMPs on cells at the surface of implant materials, the present experiments aimed to describe effects of BMP-2 on primary cultures derived from calvarial fragments of neonatal rats. The cells were stimulated with 50 nM BMP-2 added to the nutrient medium for 3 or 6 days. Light- and electronmicroscopical studies showed that cells in the sprouting zones were larger and more often spindle shaped. Stimulated cells had more nucleoli than control cells and the endoplasmic reticulum was widened. They retained properties of typical bone cells: An immunhistochemical analysis showed that stimulated cells increased the activity of alkaline phosphatase, they secreted collagen type I and to a minor extent collagen type III. In BMP-2 treated cells the pattern of cells stained for actin, desmin and vimentin hardly changed whereas extracellular fibronectin appeared to be less cross-linked in BMP-2 treated cultures. The distribution and labeling strength of osteocalcin, a specific marker protein of bone cells did not change markedly. After exposure to BMP-2 cells tended to detach from the cover slips. Electron microscopy showed a reduced number of cell processes possibly facilitating the detachment and/or mobility. Stimulated cells contained an increased number of lamellar bodies which may reflect an increased synthesis and/or membrane turnover. Staining of non-osseous cells with anti-CD68-or anti-myeloid antibodies revealed that the small percentage of these cells regularly occurring in primary cultures

  7. Neuronal-like differentiation of bone marrow-derived mesenchymal stem cells induced by striatal extracts from a rat model of Parkinson's disease

    Institute of Scientific and Technical Information of China (English)

    Xiaoling Qin; Wang Han; Zhigang Yu

    2012-01-01

    A rat model of Parkinson's disease was established by 6-hydroxydopamine injection into the medial forebrain bundle. Bone marrow-derived mesenchymal stem cells (BMSCs) were isolated from the femur and tibia, and were co-cultured with 10% and 60% lesioned or intact striatal extracts. The results showed that when exposed to lesioned striatal extracts, BMSCs developed bipolar or multi-polar morphologies, and there was an increase in the percentage of cells that expressed glial fibrillary acidic protein (GFAP), nestin and neuron-specific enolase (NSE). Moreover, the percentage of NSE-positive cells increased with increasing concentrations of lesioned striatal extracts. However, intact striatal extracts only increased the percentage of GFAP-positive cells. The findings suggest that striatal extracts from Parkinson's disease rats induce BMSCs to differentiate into neuronal-like cells in vitro.

  8. Bone marrow transplantation restores epidermal basement membrane protein expression and rescues epidermolysis bullosa model mice

    OpenAIRE

    Fujita, Yasuyuki; Abe, Riichiro; Inokuma, Daisuke; Sasaki, Mikako; Hoshina, Daichi; Natsuga, Ken; Nishie, Wataru; McMillan, James R.; Nakamura, Hideki; Shimizu, Tadamichi; Akiyama, Masashi; Sawamura, Daisuke; Shimizu, Hiroshi

    2010-01-01

    Attempts to treat congenital protein deficiencies using bone marrow-derived cells have been reported. These efforts have been based on the concepts of stem cell plasticity. However, it is considered more difficult to restore structural proteins than to restore secretory enzymes. This study aims to clarify whether bone marrow transplantation (BMT) treatment can rescue epidermolysis bullosa (EB) caused by defects in keratinocyte structural proteins. BMT treatment of adult collagen XVII (Col17) ...

  9. Pinto Bean Hull Extract Supplementation Favorably Affects Markers of Bone Metabolism and Bone Structure in Mice

    Science.gov (United States)

    Dry edible beans (Phaseolus vulgaris) have many health benefits attributed to their high content of protein, non-digestible starches, fiber, and other bioactive components. Hulls from dry beans are rich in phenolics known to possess antioxidant activity that is beneficial to human health. The object...

  10. A new heterologous fibrin sealant as scaffold to recombinant human bone morphogenetic protein-2 (rhBMP-2) and natural latex proteins for the repair of tibial bone defects.

    Science.gov (United States)

    Machado, Eduardo Gomes; Issa, João Paulo Mardegan; Figueiredo, Fellipe Augusto Tocchini de; Santos, Geovane Ribeiro Dos; Galdeano, Ewerton Alexandre; Alves, Mariana Carla; Chacon, Erivelto Luis; Ferreira Junior, Rui Seabra; Barraviera, Benedito; Cunha, Marcelo Rodrigues da

    2015-04-01

    Tissue engineering has special interest in bone tissue aiming at future medical applications Studies have focused on recombinant human bone morphogenetic protein-2 (rhBMP-2) and natural latex proteins due to the osteogenic properties of rhBMP-2 and the angiogenic characteristic of fraction 1 protein (P-1) extracted from the rubber tree Hevea brasiliensis. Furthermore, heterologous fibrin sealant (FS) has been shown as a promising alternative in regenerative therapies. The aim of this study was to evaluate these substances for the repair of bone defects in rats. A bone defect measuring 3mm in diameter was created in the proximal metaphysis of the left tibia of 60 rats and was implanted with rhBMP-2 or P-1 in combination with a new heterologous FS derived from snake venom. The animals were divided into six groups: control (unfilled bone defect), rhBMP-2 (defect filled with 5μg rhBMP-2), P-1 (defect filled with 5μg P-1), FS (defect filled with 8μg FS), FS/rhBMP-2 (defect filled with 8μg FS and 5μg rhBMP-2), FS/P-1 (defect filled with 8μg FS and 5μg P-1). The animals were sacrificed 2 and 6 weeks after surgery. The newly formed bone projected from the margins of the original bone and exhibited trabecular morphology and a disorganized arrangement of osteocyte lacunae. Immunohistochemical analysis showed intense expression of osteocalcin in all groups. Histometric analysis revealed a significant difference in all groups after 2 weeks (p0.05). A statistically significant difference (p<0.05) was observed in all groups after 6 weeks in relation to the volume of newly formed bone in the surgical area. In conclusion, the new heterologous fibrin sealant was found to be biocompatible and the combination with rhBMP-2 showed the highest osteogenic and osteoconductive capacity for bone healing. These findings suggest a promising application of this combination in the regeneration surgery. PMID:25825118

  11. An improved protein extraction method for detecting ruminant material in feed using lateral flow device dipsticks

    Directory of Open Access Journals (Sweden)

    Karanam, M.

    2011-01-01

    Full Text Available The currently used immunological lateral flow based dipstick technology is not sensitive to detect the presence of ruminant animal material below 1% level of contamination in feed. The possible reason is that the processed animal proteins are subjected to chemicals and to high temperatures during the rendering process, thereby leading to either their denaturation or degradation. Also some feed samples producing viscous extracts or containing high contents of mineral or vitamins delay the migration flow in the test device and may thus give rise to either inconclusive or false positive results. Veterinary Laboratory Agency focused its efforts on developing a new protein extraction method through the EU Safeed-PAP project with the aim to increase the sensitivity of the dipstick method. The method developed is based on a bone sedimentation procedure coupled with an EDTA digestion step in order to liberate the protected proteins which are covalently bound to mineral structures within the bone mineral cage. All the results using this method showed that the feed contaminated with ruminant material is detected at 0.1 to 0.2% levels.

  12. Extractable protein of radiation vulcanized natural rubber latex

    Energy Technology Data Exchange (ETDEWEB)

    Soebianto, Y.S. [Center for Research and Development of Isotopes and Radiation Technology, BATAN, Jakarta (Indonesia); Upul, R.M. [Rubber Research Institute of Sri Lanka, Ratmalana (Sri Lanka); Makuuchi, K.; Yoshii, F.; Kume, T. [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-03-01

    A new method to reduce the protein level in the latex products by irradiation is reported. Water soluble protein (WSP) solution (10%) was added into radiation vulcanized NR latex (RVNRL) as much as 3 phr in three different processes: added to RVNRL, added to re-centrifuged RVNRL, and added to RVNRL followed by centrifugation. The protein content was determined by enhanced BCA method, and identified by SDS-PAGE analysis. Addition of WSP followed by centrifugation reduces EP up to the minimum protein detection, and shortens the leaching time to 20-30 min. SDS-PAGE analysis confirms the reduction of soluble protein in the serum phase, and disappearance of protein bands in the rubber extract. Protein-WSP interaction produces water soluble complex, and removed by centrifugation. The molecular weight of WSP dictates the efficiency of protein removal. (author)

  13. Extractable protein of radiation vulcanized natural rubber latex

    International Nuclear Information System (INIS)

    A new method to reduce the protein level in the latex products by irradiation is reported. Water soluble protein (WSP) solution (10%) was added into radiation vulcanized NR latex (RVNRL) as much as 3 phr in three different processes: added to RVNRL, added to re-centrifuged RVNRL, and added to RVNRL followed by centrifugation. The protein content was determined by enhanced BCA method, and identified by SDS-PAGE analysis. Addition of WSP followed by centrifugation reduces EP up to the minimum protein detection, and shortens the leaching time to 20-30 min. SDS-PAGE analysis confirms the reduction of soluble protein in the serum phase, and disappearance of protein bands in the rubber extract. Protein-WSP interaction produces water soluble complex, and removed by centrifugation. The molecular weight of WSP dictates the efficiency of protein removal. (author)

  14. Physiochemical characterizations of hydroxyapatite extracted from bovine bones by three different methods: Extraction of biologically desirable HAp

    International Nuclear Information System (INIS)

    In the present study, subcritical water and alkaline hydrolysis methods are proposed methodologies for extraction of natural hydroxyapatite bioceramic from bovine bone. In these processes, the bovine bones powder were treated by high pressure water at 250 deg. C for 1 h and 25% (wt) sodium hydroxide at 250 deg. C for 5 h, respectively. Also the conventional calcination methodology has been utilized as well (T = 850 deg. C for 1 h). The obtained apatites from the three treatment processes have been characterized by powder X-ray diffraction analysis (XRD), Fourier transform infrared spectroscopy (FT IR), transmission electron microscopy (TEM), thermal gravimetric analysis (TGA), electron scanning microscopy (SEM), energy dispersive X-ray analysis (EDX) and field emission scanning electron microscopy (FE SEM). FT IR and XRD results affirmed that both the proposed methods and the traditional one can eliminate the collagen and other organic materials present in the bovine bones. The physiochemical characterizations for the obtained apatites have proved that the subcritical water and the alkaline hydrolysis relatively preserve the carbonate content present in the biological apatite, so they yield carbonated hydroxyapatite which is medically preferable. While, the thermal process produces almost hydroxyapatite carbonate-free

  15. Effect of protein malnutrition on the metabolism of bone collagen in albino rats

    International Nuclear Information System (INIS)

    The effect of protein malnutrition on the metabolism of collagen in bone was studied in young female albino rats after a single injection of 3H-proline. Both specific and total radioactivities of hydroxyproline in the total collagen of the bone were found to decrease in the protein-deficient animals, indicating decreased rate of collagen synthesis. In the urine the amount of hydroxyproline excreted and total radioactivity of 3H-hydroxyproline were greatly decreased. The results of the present investigation therefore clearly indicate decreased synthesis and catabolism of collagen in bones of protein deficient animals compared to controls. (auth.)

  16. Protein PocketViewer: A Web-Service Based Interface for Protein Pocket Extraction and Visualization

    OpenAIRE

    Xiaoyu Zhang; Martin Gordon

    2013-01-01

    One important problem in bioinformatics is to study pockets or tunnels within the protein structure. These pocket or tunnel regions are significant because they indicate areas of ligand binding or enzymatic reactions, and tunnels are often solvent ion conductance areas. The Protein Pocket Viewer (PPV) is a web interface that allows the user to extract and visualize the protein pockets in a browser, based on the algorithm in [1]. The PPV packaged the pocket extraction executable as a web servi...

  17. Short term effects on bone quality associated with consumption of soy protein isolate and other dietary protein sources in rapidly growing female rats

    Science.gov (United States)

    Beneficial effects of soy protein consumption on bone quality have been reported. The effects of other dietary protein sources such as whey protein hydrolysate (WPH) and rice protein isolate (RPI) on bone growth has been less well examined. The current study compared effects of feeding soy protein i...

  18. Mesenchymal stromal cells from bone marrow treated with bovine tendon extract acquire the phenotype of mature tenocytes☆

    Science.gov (United States)

    Augusto, Lívia Maria Mendonça; Aguiar, Diego Pinheiro; Bonfim, Danielle Cabral; dos Santos Cavalcanti, Amanda; Casado, Priscila Ladeira; Duarte, Maria Eugênia Leite

    2016-01-01

    Objective This study evaluated in vitro differentiation of mesenchymal stromal cells isolated from bone marrow, in tenocytes after treatment with bovine tendon extract. Methods Bovine tendons were used for preparation of the extract and were stored at −80 °C. Mesenchymal stromal cells from the bone marrow of three donors were used for cytotoxicity tests by means of MTT and cell differentiation by means of qPCR. Results The data showed that mesenchymal stromal cells from bone marrow treated for up to 21 days in the presence of bovine tendon extract diluted at diminishing concentrations (1:10, 1:50 and 1:250) promoted activation of biglycan, collagen type I and fibromodulin expression. Conclusion Our results show that bovine tendon extract is capable of promoting differentiation of bone marrow stromal cells in tenocytes. PMID:26962503

  19. Bone morphogenetic protein 4 mediates human embryonic germ cell derivation.

    Science.gov (United States)

    Hiller, Marc; Liu, Cyndi; Blumenthal, Paul D; Gearhart, John D; Kerr, Candace L

    2011-02-01

    Human primordial germ cells (PGCs) have proven to be a source of pluripotent stem cells called embryonic germ cells (EGCs). Unlike embryonic stem cells, virtually little is known regarding the factors that regulate EGC survival and maintenance. In mice, the growth factor bone morphogenetic protein 4 (BMP4) has been shown to be required for maintaining mouse embryonic stem cells, and disruptions in this gene lead to defects in mouse PGC specification. Here, we sought to determine whether recombinant human BMP4 could influence EGC derivation and/or human PGC survival. We found that the addition of recombinant BMP4 increased the number of human PGCs after 1 week of culture in a dose-responsive manner. The efficiency of EGC derivation and maintenance in culture was also enhanced by the presence of recombinant BMP4 based on alkaline phosphatase and OCT4 staining. In addition, an antagonist of the BMP4 pathway, Noggin, decreased PGC proliferation and led to an increase in cystic embryoid body formation. Quantitative real-time (qRT)-polymerase chain reaction analyses and immunostaining confirmed that the constituents of the BMP4 pathway were upregulated in EGCs versus PGCs. Downstream activators of the BMP4 pathway such as ID1 and phosphorylated SMADs 1 and 5 were also expressed, suggesting a role of this growth factor in EGC pluripotency. PMID:20486775

  20. The influence of dietary crude protein intake on bone and mineral metabolism in sheep

    Directory of Open Access Journals (Sweden)

    T.S. Brand

    1999-07-01

    Full Text Available Increased dietary protein consumption is thought to cause calciuresis, a negative calcium balance and increased bone loss that may result in skeletal deformities and fracture. To explore this hypothesis, 40 approximately 100-day-old meat-type Merino ram lambs were fed, for 6 months, diets with an increasing crude protein (CP content (114, 142, 171 and 190 g/kg DM but approximately on an iso-nutrient basis with regard to metabolisable energy, calcium and phosphorus. Increased protein consumption modestly (NS enhanced calciuresis and resulted in significant (P < 0.01 limb skewness. This could not, however, be ascribed to osteopaenic bones, and compared with animals consuming lower protein rations, the bone mineral density (BMD and vertebral trabecular bone volume of animals fed high protein diets were significantly increased: theBMDof thoracic vertebrae was positively related to the CP intake (r=0.62; P < 0.001. In animals consuming higher protein diets, skeletal radiology and quantitative bone histology revealed no evidence of increased bone turnover as would be expected in animals that are in negative calcium balance. No relationship existed between limb skewness and the growth rate of lambs. However, the ratio of Ca:P in the forelimb (r = -0.98, vertebrae (r = -0.72 and rib (r = -0.42 was found to be inversely correlated with increased protein intake and resulted from an increase in the phosphorus content of bone, while the amount of bone calcium was unaffected. We conclude that qualitative micro-architectural abnormalities, and not mere bone loss, may underlie the skeletal deformities induced by increased protein consumption in sheep.

  1. Extractable protein of radiation vulcanized natural rubber latex

    International Nuclear Information System (INIS)

    Protein remained in the latex products are reported to cause serious allergy. A new method to reduce the protein level in the latex products by irradiation is reported. Water soluble protein (WSP) solution (10%) was added into radiation vulcanized NR latex (RVNRL) in three different processes. The amount of WSP was 3 phr. It was only added to RVNRL (standard), added to re-centrifuged RVNRL (pre-centrifugation), and added to RVNRL followed by centrifugation (post-centrifugation). The protein content was determined by enhanced BCA method, and identified by SDS-PAGE. Extractable protein (EP) from the rubber has been reduced up to the minimum protein detection by combining WSP addition and centrifugation. Short leaching time (20-30 min.) can be achieved after the combine treatment, and SDS-PAGE confirms the reduction of soluble protein in the serum phase, and disappearance of protein bands in the rubber extract. Protein-WSP interaction produces water soluble complex, and removed by centrifugation. The efficiency of protein removal by WSP depends on its molecular weight of WSP which relates to its water solubility. (author)

  2. Experimental study on bone mineral determination in the wound healing process after tooth extraction by 125I-photon absorptiometry

    International Nuclear Information System (INIS)

    A study was carried out by 125I-photon absorptiometry in tooth extraction wounds of dogs to determine the bone mineral content of the mandibular premolar region. The bone mineral content of excised mandibles was measured on the 3rd, 7th, 14th, 21st, 28th, 56th, and the 84th day, respectively, after tooth extraction. These were then compared with the values obtained by photodensitometry and by histological findings. The bone mineral content obtained by 125I-photon absorptiometry varied within a range of 0.270 and 0.720 g/cm2, over a period of 84-days. The highest mineral content value was noted in the apical site. Lower values were noted in the central site and with still lower values in the marginal site. In each case the bone mineral content which deacreased on the 3rd and 7th day after extraction tended to increase gradually thereafter. On the 56th and 84th day after tooth extraction, an increase in bone mineral in the margical site and a deacrease in the apical site was observed. This observation indicated that homogenization of the bone mineral occurred in the tooth extraction wound. Bone changes in the histological findings related to the healing process and the changes of values obtained by photodensitometry coincided with the changes shown in 125I-photon absorptiometry. (author)

  3. DNA in ancient bone - where is it located and how should we extract it?

    Science.gov (United States)

    Campos, Paula F; Craig, Oliver E; Turner-Walker, Gordon; Peacock, Elizabeth; Willerslev, Eske; Gilbert, M Thomas P

    2012-01-20

    Despite the widespread use of bones in ancient DNA (aDNA) studies, relatively little concrete information exists in regard to how the DNA in mineralised collagen degrades, or where it survives in the material's architecture. While, at the macrostructural level, physical exclusion of microbes and other external contaminants may be an important feature, and, at the ultrastructural level, the adsorption of DNA to hydroxyapatite and/or binding of DNA to Type I collagen may stabilise the DNA, the relative contribution of each, and what other factors may be relevant, are unclear. There is considerable variation in the quality of DNA retrieved from bones and teeth. This is in part due to various environmental factors such as temperature, proximity to free water or oxygen, pH, salt content, and exposure to radiation, all of which increase the rate of DNA decay. For example, bone specimens from sites at high latitudes usually yield better quality DNA than samples from temperate regions, which in turn yield better results than samples from tropical regions. However, this is not always the case, and rates of success of DNA recovery from apparently similar sites are often strikingly different. The question arises as to whether this may be due to post-collection preservation or just an artefact of the extraction methods used in these different studies? In an attempt to resolve these questions, we examine the efficacy of DNA extraction methods, and the quality and quantity of DNA recovered from both artificially degraded, and genuinely ancient, but well preserved, bones. In doing so we offer hypotheses relevant to the DNA degradation process itself, and to where and how the DNA is actually preserved in ancient bone. PMID:21855309

  4. The osteogenic differentiation stimulating activity of Sea cucumber methanolic crude extraction on rat bone marrow mesenchymal stem cells

    OpenAIRE

    Javad Baharara; Elaheh Amini; Mohammad Amin Kerachian; Mozhgan Soltani

    2014-01-01

    Objective(s): Sea cucumber derived bioactive compound is considered efficient in treatment of bone disorders. This study was performed to evaluate the effect of this extract on differentiation of rat bone marrow mesenchymal stem cells (rBMMSc) into osteogenic lineage. Materials and Methods: Isolated rBMMSc were grown in DMEM supplemented with 10% FBS. The cells were exposed to different concentration of extract. After 21 days, Alizarin red staining, alkaline phosphatase assay and RT-PCR were ...

  5. The role of bone marrow-derived cells in bone fracture repair in a green fluorescent protein chimeric mouse model

    International Nuclear Information System (INIS)

    We investigated the role of bone marrow cells in bone fracture repair using green fluorescent protein (GFP) chimeric model mice. First, the chimeric model mice were created: bone marrow cells from GFP-transgenic C57BL/6 mice were injected into the tail veins of recipient wild-type C57BL/6 mice that had been irradiated with a lethal dose of 10 Gy from a cesium source. Next, bone fracture models were created from these mice: closed transverse fractures of the left femur were produced using a specially designed device. One, three, and five weeks later, fracture lesions were extirpated for histological and immunohistochemical analyses. In the specimens collected 3 and 5 weeks after operation, we confirmed calluses showing intramembranous ossification peripheral to the fracture site. The calluses consisted of GFP- and osteocalcin-positive cells at the same site, although the femur consisted of only osteocalcin-positive cells. We suggest that bone marrow cells migrated outside of the bone marrow and differentiated into osteoblasts to make up the calluses

  6. Recombinant human bone morphogenetic protein-2 in debridement and impacted bone graft for the treatment of femoral head osteonecrosis.

    Directory of Open Access Journals (Sweden)

    Wei Sun

    Full Text Available The purpose of this study was to compare the clinical outcomes of impacted bone graft with or without recombinant human bone morphogenetic protein-2 (rhBMP-2 for osteonecrosis of the femoral head (ONFH. We examined the effect of bone-grafting through a window at the femoral head-neck junction, known as the "light bulb" approach, for the treatment of ONFH with a combination of artificial bone (Novobone mixed with or without rhBMP-2. A total of 42 patients (72 hips were followed-up from 5 to 7.67 years (average of 6.1 years. The patients with and without BMP were the first group (IBG+rhBMP-2 and the second group (IBG, respectively. The clinical effectiveness was evaluated by Harris hip score (HHS. The radiographic follow-up was evaluated by pre-and postoperative X-ray and CT scan. Excellent, good, and fair functions were obtained in 36, 12, and 7 hips, respectively. The survival rate was 81.8% and 71.8% in the first and second group, respectively. However, the survival rate was 90.3% in ARCO stage IIb, c, and only 34.6% in ARCO stage IIIa (P<0.05. It was concluded that good and excellent mid-term follow-up could be achieved in selected patients with ONFH treated with impacted bone graft operation. The rhBMP-2 might improve the clinical efficacy and quality of bone repair.

  7. Relationship of Circulating Total Homocysteine and C-Reactive Protein to Trabecular Bone in Postmenopausal Women

    Science.gov (United States)

    Homocysteine (Hcy) and C-reactive protein (CRP) are novel risk factors for osteoporosis. The purpose of this analysis was to determine the relationship of Hcy and CRP to volumetric trabecular bone, but also to assess their relationship to areal composite bone in healthy postmenopausal women (N=184)....

  8. Extracting protein regulatory networks with graphical models.

    Science.gov (United States)

    Grzegorczyk, Marco

    2007-09-01

    During the last decade the development of high-throughput biotechnologies has resulted in the production of exponentially expanding quantities of biological data, such as genomic and proteomic expression data. One fundamental problem in systems biology is to learn the architecture of biochemical pathways and regulatory networks in an inferential way from such postgenomic data. Along with the increasing amount of available data, a lot of novel statistical methods have been developed and proposed in the literature. This article gives a non-mathematical overview of three widely used reverse engineering methods, namely relevance networks, graphical Gaussian models, and Bayesian networks, whereby the focus is on their relative merits and shortcomings. In addition the reverse engineering results of these graphical methods on cytometric protein data from the RAF-signalling network are cross-compared via AUROC scatter plots. PMID:17893851

  9. Extraction and characterisation of apatite- and tricalcium phosphate-based materials from cod fish bones

    International Nuclear Information System (INIS)

    Apatite- and tricalcium phosphate-based materials were produced from codfish bones, thus converting a waste by-product from the food industry into high added-valued compounds. The bones were annealed at temperatures between 900 and 1200 °C, giving a biphasic material of hydroxyapatite and tricalcium phosphate (Ca10(PO4)6(OH)2 and β-Ca(PO4)3) with a molar proportion of 75:25, a material widely used in biomedical implants. The treatment of the bones in solution prior to their annealing changed the composition of the material. Single phase hydroxyapatite, chlorapatite (Ca10(PO4)6Cl2) and fluorapatite (Ca10(PO4)6F2) were obtained using CaCl2 and NaF solutions, respectively. The samples were analysed by several techniques (X-ray diffraction, infrared spectroscopy, scanning electron microscopy and differential thermal/thermogravimetric analysis) and by elemental analyses, to have a more complete understanding of the conversion process. Such compositional modifications have never been performed before for these materials of natural origin to tailor the relative concentrations of elements. This paper shows the great potential for the conversion of this by-product into highly valuable compounds for biomedical applications, using a simple and effective valorisation process. - Highlights: ► Apatite and calcium phosphate compounds extraction from cod fish bonesBone calcination: biphasic material hydroxyapatite-calcium phosphate production ► Bone pre-treatments in solution change the material composition. ► Single phase materials (hydroxy-, chloro- or fluoroapatite) are obtained. ► Concentration of other elements (Na, F, Cl) suitable for biomedical applications

  10. Extraction and characterisation of apatite- and tricalcium phosphate-based materials from cod fish bones

    Energy Technology Data Exchange (ETDEWEB)

    Piccirillo, C.; Silva, M.F. [CBQF/Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, Porto (Portugal); Pullar, R.C. [Dept. Engenharia de Materiais e Ceramica/CICECO, Universidade de Aveiro, Aveiro (Portugal); Braga da Cruz, I. [CBQF/Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, Porto (Portugal); WeDoTech, CiDEB/Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, Porto (Portugal); Jorge, R. [WeDoTech, CiDEB/Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, Porto (Portugal); Pintado, M.M.E. [CBQF/Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, Porto (Portugal); Castro, P.M.L., E-mail: plcastro@porto.ucp.pt [CBQF/Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, Porto (Portugal)

    2013-01-01

    Apatite- and tricalcium phosphate-based materials were produced from codfish bones, thus converting a waste by-product from the food industry into high added-valued compounds. The bones were annealed at temperatures between 900 and 1200 Degree-Sign C, giving a biphasic material of hydroxyapatite and tricalcium phosphate (Ca{sub 10}(PO{sub 4}){sub 6}(OH){sub 2} and {beta}-Ca(PO{sub 4}){sub 3}) with a molar proportion of 75:25, a material widely used in biomedical implants. The treatment of the bones in solution prior to their annealing changed the composition of the material. Single phase hydroxyapatite, chlorapatite (Ca{sub 10}(PO{sub 4}){sub 6}Cl{sub 2}) and fluorapatite (Ca{sub 10}(PO{sub 4}){sub 6}F{sub 2}) were obtained using CaCl{sub 2} and NaF solutions, respectively. The samples were analysed by several techniques (X-ray diffraction, infrared spectroscopy, scanning electron microscopy and differential thermal/thermogravimetric analysis) and by elemental analyses, to have a more complete understanding of the conversion process. Such compositional modifications have never been performed before for these materials of natural origin to tailor the relative concentrations of elements. This paper shows the great potential for the conversion of this by-product into highly valuable compounds for biomedical applications, using a simple and effective valorisation process. - Highlights: Black-Right-Pointing-Pointer Apatite and calcium phosphate compounds extraction from cod fish bones Black-Right-Pointing-Pointer Bone calcination: biphasic material hydroxyapatite-calcium phosphate production Black-Right-Pointing-Pointer Bone pre-treatments in solution change the material composition. Black-Right-Pointing-Pointer Single phase materials (hydroxy-, chloro- or fluoroapatite) are obtained. Black-Right-Pointing-Pointer Concentration of other elements (Na, F, Cl) suitable for biomedical applications.

  11. The Inhibiting Effect of Bone Protein Hydrolysates on Lipid Oxidation in Pork Patties

    Institute of Scientific and Technical Information of China (English)

    DIAO Jingjing; DIAO Xinping; KONG Baohua; CHEN Hongsheng

    2009-01-01

    Bone protein hydrolysates were prepared by limited alcalase hydrolysis (5 h). The hydrolysates were formulated (0-3%,w/w) into pork patties to determine the antioxidant efficacy. 0.02% BHA (butylated hydroxyanisole) was used as a positive control.Lipid oxidation in patties during storage was analyzed by measuring the TBARS and protein carbonyl content. The results showed that bone protein hydrolysates possessed significant antioxidant activity, and antioxidant activity increased with the increasing hydrolysates concentration. Sensory evaluation indicated that bone protein hydrolysates improved the color and decreased lipid oxidation flavor of pork patties. The 2% bone hydrolysates possessed the highest antioxidant activity and better sensory quality, and its effect was closed to 0.02% BHA.

  12. Skin, bone and muscle collagen extraction from the trash fish, leather jacket (Odonus niger) and their characterization

    OpenAIRE

    Muralidharan, Nagarajan; Jeya Shakila, Robinson; Sukumar, Durairaj; Jeyasekaran, G.

    2011-01-01

    Acid soluble (ASC) and pepsin soluble (PSC) collagens were extracted from the skin, bone and muscle of a trash fish, leather jacket (Odonus niger) by three different extraction methods. Method I gave 46–50% yield for ASC, Method II gave 49–58% yield for both ASC and PSC and Method III gave 64–71% yield for PSC. The addition of pepsin had increased the yield by 30–45%. The yields of collagen from skin and bone were higher than muscle. SDS-PAGE pattern revealed that skin and bone collagen as Ty...

  13. Ethanol Extract of Atractylodes macrocephala Protects Bone Loss by Inhibiting Osteoclast Differentiation

    Directory of Open Access Journals (Sweden)

    Youn-Hwan Hwang

    2013-06-01

    Full Text Available The rhizome of Atractylodes macrocephala has been used mainly in Traditional Chinese Medicine for invigorating the functions of the stomach and spleen. In the present study, we investigated the inhibitory effect of the 70% ethanol extract of the rhizome of Atractylodes macrocephala (AMEE on osteoclast differentiation. We found that AMEE inhibits osteoclast differentiation from its precursors induced by receptor activator of nuclear factor-κB ligand (RANKL, an essential cytokine required for osteoclast differentiation. AMEE attenuated RANKL-induced activation of NF-κB signaling pathway, subsequently inhibiting the induction of osteoclastogenic transcription factors, c-Fos and nuclear factor of activated T cells cytoplasmic 1. Consistent with the in vitro results, administration of AMEE protected RANKL-induced bone loss in mice. We also identified atractylenolide I and II as active constituents contributing to the anti-osteoclastogenic effect of AMEE. Taken together, our results demonstrate that AMEE has a protective effect on bone loss via inhibiting osteoclast differentiation and suggest that AMEE may be useful in preventing and treating various bone diseases associated with excessive bone resorption.

  14. Multiplex Assay for Protein Profiling and Potency Measurement of German Cockroach Allergen Extracts.

    Directory of Open Access Journals (Sweden)

    Taruna Khurana

    Full Text Available German cockroach (GCr allergens induce IgE responses and may cause asthma. Commercial GCr allergen extracts are variable and existing assays may not be appropriate for determining extract composition and potency.Our aim was to develop a multiplex antibody/bead-based assay for assessment of GCr allergen extracts.Single chain fragment variable (scFv antibodies against GCr were obtained by screening libraries derived from naïve human lymphocytes and hyperimmunized chicken splenocytes and bone marrow. Selected clones were sequenced and characterized by immunoblotting. Eighteen scFv antibodies (17 chicken, 1 human coupled to polystyrene beads were used in this suspension assay; binding of targeted GCr allergens to antibody-coated beads was detected using rabbit antisera against GCr, and against specific allergens rBla g 1, rBla g 2, and rBla g 4. The assay was tested for specificity, accuracy, and precision. Extracts were also compared by IgE competition ELISA.Chicken scFv's generated eight different binding patterns to GCr proteins from 14 to 150 kDa molecular weight. Human scFv's recognized a 100 kDa GCr protein. The multiplex assay was found to be specific and reproducible with intra-assay coefficient of variation (CV of 2.64% and inter-assay CV of 10.0%. Overall potencies of various GCr extracts were calculated using mean logEC50s for eight selected scFvs. Overall potency measures were also analyzed by assessing the contributions to potency of each target.An scFv antibody-based multiplex assay has been developed capable of simultaneously measuring different proteins in a complex mixture, and to determine the potencies and compositions of allergen extracts.

  15. Antigen-free bovine cancellous bone loaded with recombinant human bone morphogenetic protein-2 for the repair of tibial bone defects in goat model.

    Science.gov (United States)

    Li, Donghai; Deng, Liqing; Yang, Zhouyuan; Xie, Xiaowei; Kang, Pengde; Tan, Zhen

    2016-04-01

    Antigen-free bovine cancellous bone has good performances of porous network structures and mechanics with antigen extracted. To develop a bioactive scaffold for enhancing bone repair and evaluate its biological property, rhBMP-2 loaded with antigen-free bovine cancellous bone was used to treat tibial bone defect. Twenty-four healthy adult goats were chosen to establish goat defects model and randomly divided into four groups. The goats were treated with rhBMP-2/antigen-free bovine cancellous bone scaffolds (group A), autogenous cancellous bone graft (group B), porous tricalciumphosphate scaffolds (group C) and nothing (group D). Animals were evaluated with radiological and histological methods at 4, 8 and 12 weeks after surgery. The gray value of radiographs was used to evaluate the healing of the defects, which revealed that the group A had a better outcome of defect healing compared with group C at 4, 8 and 12 weeks, respectively (p difference between groups A and B was without significance at each time (p > 0.05). The newly formed bone area was calculated from histological sections, and the results indicated that the amount of new bone in group A increased significantly compared with that in group C (p  0.05) at 4, 8 and 12 weeks, respectively. In addition, the expression of collagen I and vascular endothelial growth factor by real-time polymerase chain reaction at 12 weeks in group A was significantly higher than that in group C (p = 0.034, p = 0.032, respectively), but no significant differences were found when compared with that in group B (p = 0.36, p = 0.54, respectively). At the same time, group C presented better results than group D on bone defects healing. Therefore, the composites of antigen-free bovine cancellous bone loaded with rhBMP-2 have a good osteoinductive activity and capacity to promote the repair of bone defects. PMID:26801475

  16. Promotion of bone growth by dietary soy protein isolate: Comparision with dietary casein, whey hydrolysate and rice protein isolate in growing female rats

    Science.gov (United States)

    The effects of different dietary protein sources(casein (CAS), soy protein isolate (SPI), whey protein hydrolysate (WPH) and rice protein isolate (RPI)) on bone were studied in intact growing female rats and in ovarectomized (OVX) rats showing sex steroid deficiency-induced bone loss. In addition, S...

  17. Parathyroid Hormone-Related Protein, Its Regulation of Cartilage and Bone Development, and Role in Treating Bone Diseases.

    Science.gov (United States)

    Martin, T John

    2016-07-01

    Although parathyroid hormone-related protein (PTHrP) was discovered as a cancer-derived hormone, it has been revealed as an important paracrine/autocrine regulator in many tissues, where its effects are context dependent. Thus its location and action in the vasculature explained decades-long observations that injection of PTH into animals rapidly lowered blood pressure by producing vasodilatation. Its roles have been specified in development and maturity in cartilage and bone as a crucial regulator of endochondral bone formation and bone remodeling, respectively. Although it shares actions with parathyroid hormone (PTH) through the use of their common receptor, PTHR1, PTHrP has other actions mediated by regions within the molecule beyond the amino-terminal sequence that resembles PTH, including the ability to promote placental transfer of calcium from mother to fetus. A striking feature of the physiology of PTHrP is that it possesses structural features that equip it to be transported in and out of the nucleus, and makes use of a specific nuclear import mechanism to do so. Evidence from mouse genetic experiments shows that PTHrP generated locally in bone is essential for normal bone remodeling. Whereas the main physiological function of PTH is the hormonal regulation of calcium metabolism, locally generated PTHrP is the important physiological mediator of bone remodeling postnatally. Thus the use of intermittent injection of PTH as an anabolic therapy for bone appears to be a pharmacological application of the physiological function of PTHrP. There is much current interest in the possibility of developing PTHrP analogs that might enhance the therapeutic anabolic effects. PMID:27142453

  18. Recombinant human bone morphogenetic protein-2 suspended in fibrin glue enhances bone formation during distraction osteogenesis in rabbits

    Science.gov (United States)

    Li, Yunfeng; Li, Rui; Hu, Jing; Song, Donghui; Jiang, Xiaowen

    2016-01-01

    Introduction Bone morphogenetic protein-2 (BMP-2) has high potential for bone formation, but its in vivo effects are unpredictable due to the short life time. This study was designed to evaluate the effects of recombinant human (rh) BMP-2 suspended in fibrin on bone formation during distraction osteogenesis (DO) in rabbits. Material and methods The in vitro release kinetics of rhBMP-2 suspended in fibrin was tested using an enzyme-linked immunosorbent assay. Unilateral tibial lengthening for 10 mm was achieved in 48 rabbits. At the completion of osteodistraction, vehicle, fibrin, rhBMP-2 or rhBMP-2 suspended in fibrin (rhBMP-2 + fibrin) was injected into the center of the lengthened gap, with 12 animals in each group. Eight weeks later, the distracted callus was examined by histology, micro-CT and biomechanical testing. Radiographs of the distracted tibiae were taken at both 4 and 8 weeks after drug treatment. Results It was found that fibrin prolonged the life span of rhBMP-2 in vitro with sustained release during 17 days. The rhBMP-2 + fibrin treated animals showed the best results in bone mineral density, bone volume fraction, cortical bone thickness by micro-CT evaluation and mechanical properties by the three-point bending test when compared to the other groups (p < 0.05). In histological images, rhBMP-2 + fibrin treatment showed increased callus formation and better gap bridging compared to the other groups. Conclusions The results of this study suggest that fibrin holds promise to be a good carrier of rhBMP-2, and rhBMP-2 suspended in fibrin showed a stronger promoting effect on bone formation during DO in rabbits. PMID:27279839

  19. Urinary deoxypyridinoline (DPD), serum bone glia protein (BGP) and bone metabolism change in hyperthyroidism

    International Nuclear Information System (INIS)

    Objective: To study the effect of thyroid function on bone metabolism. Methods: Urinary DPD, Serum FT3, FT4 and BGP levels were determined with chemiluminescence assay and RIA in 41 patients with hyperthyroidism and 47 healthy controls. Results: Urinary DPD and serum FT3, FT4, BGP levels were significantly higher in patients with hyperthyroidism than those in healthy controls (p < 0.01). Conclusion: The data showed that hyperthyroidism was correlated with bone metabolism

  20. Palatability of water-soluble extracts of protein sources and replacement of fishmeal by a selected mixture of protein sources for juvenile turbot ( Scophthalmus maximus)

    Science.gov (United States)

    Dong, Chun; He, Gen; Mai, Kangsen; Zhou, Huihui; Xu, Wei

    2016-06-01

    Poor palatability is a limiting factor for replacing fishmeal with other protein sources in aquaculture. The water-soluble molecules with low molecular weights are the major determinants of the palatability of diets. The present study was conducted to investigate the palatability of water-soluble extracts from single protein source (single extract pellets) and the mixture of these extracts with different proportions (blended extract pellets) in juvenile turbot ( Scophthalmus maximus). Then according to the palatability of blended extract pellets, an optimal mixture proportion was selected, and a new protein source made from raw protein materials with the selected proportion was formulated to replace fishmeal. Summarily, the palatability of single extract pellets for turbot was descendent from fishmeal to pet-food grade poultry by-product meal, wheat gluten meal, soybean meal, peanut meal, meat and bone meal, and corn gluten meal. Subsequently, according to the palatability of single extract pellets, 52 kinds of blended extract pellets were designed to test their palatability. The results showed that the pellets presented remarkably different palatability, and the optimal one was diet 52 (wheat gluten meal: pet-food grade poultry by-product meal: meat and bone meal: corn gluten meal = 1:6:1:2). The highest ingestion ratio (the number of pellets ingested/the number of pellets fed) was 0.73 ± 0.03, which was observed in Diet 52. Then five isonitrogenous (52% crude protein) and isocaloric (20 kJ g-1 gross energy) diets were formulated by replacing 0 (control), 35%, 50%, 65% and 80% of fishmeal with No.52 blending proportion. After a 10-weeks feeding trial, a consistent feed intake was found among all replacement treatments. Replacement level of fishmeal up to 35% did not significantly influence final body weight, specific growth rate, feed efficiency ratio, and protein efficiency ratio of turbot. Therefore, the water-soluble extracts of protein sources play an

  1. Recent Advances in Protein Extraction and Chiral Separation of Biomolecules

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Reverse micelles create unique environment in organic media. They are capable of solubilizing hydrophilic biomolecules (e.g., proteins, peptides, amino acids, and DNAs) in their aqueous interior. This feature brings about the practical use of biomaterials in organic media because reverse micelles solubilize them with the intrinsic activity. In this paper, we focus on recent two topics concerning protein extraction and chiral separation of biomolecules using liquid membranes. In the first topic, we present recent attempts to extract proteins from an aqueous solution into isooctane using reverse micelles, and some important operational parameters to achieve an efficient protein transfer are discussed. Furthermore, novel function of reverse micelles as a protein activation medium is introduced. In the reverse micellar phase, denatured proreins were completely reactivated in the reverse micellar solution. The reverse micellar technique is found to be a useful tool not only for protein separation but also for protein refolding. Furthermore, we found that a cyclic ligand carixarene has an extraction ability to set up optimum conditions for protein transfer. In the second topic, we have found that a supported liquid membrane (SLM) encapsulating enzymes shows high enantioselectivity (enantioselective excess value is over 96%) in the transport of racemic pharmaceutical compound ibuprofen. A different experiment also suggests that the α-chymotrypsin-catalyzed reactions droved the enantioselective transport of L-phenylalanine based on the enantioselectivity of the enzyme. The SLM encapsulating the surfactant-enzyme complex enabled the highly enantioselective separation of racemic mixtures. It can be envisioned that arrangement of appropriate enzymes in the SLM system will allow enantioselective separation of various useful organic compounds.

  2. High Protein Intake Improves Insulin Sensitivity but Exacerbates Bone Resorption in Immobility (WISE Study)

    Science.gov (United States)

    Heer, Martina; Smith, Scott M.; Frings-Meuthen, Petra; Zwart, Sara R.; Baecker, Natalie

    2012-01-01

    Inactivity, like bed rest (BR), causes insulin resistance (IR) and bone loss even in healthy subjects. High protein intake seems to mitigate this IR but might exacerbate bone loss. We hypothesized that high protein intake (animal:vegetable protein ratio: 60:40), isocaloric, compared to the control group plus high potassium intake would prevent IR without affecting bone turnover. After a 20-day ambulatory adaptation to controlled confinement and diet, 16 women participated in a 60-day, 6 deg head-down-tilt BR and were assigned randomly to one of the two groups. Control subjects (CON, n=8) received 1g/kg body mass/d dietary protein. Nutrition subjects (NUT, n=8) received 1.45g/kg body mass/d dietary protein plus 7.2g branched chain amino acids per day during BR. All subjects received 1670 kcal/d. Bed rest decreased glucose disposal by 35% (pprotein intake prevented insulin resistance, but exacerbated bed rest induced increase in bone resorption markers C-telopeptide (> 30%) and Ntelopeptide (>20%) (both: pprotein intake. We conclude from these results that high protein intake might positively affect glucose tolerance, but might also foster bone loss. Further long-duration studies are mandatory before high protein intake for diabetic patients, who have an increased fracture risk, might be recommended.

  3. PRELIMINARY STUDY OF EXTRACTABLE PROTEIN BINDING USING MALEIC ANHYDRIDE COPOLYMER

    Institute of Scientific and Technical Information of China (English)

    Thirawan Nipithakul; Ladawan Watthanachote; Nanticha Kalapat

    2012-01-01

    A preliminary study of using maleic anhydride copolymer for protein binding has been carried out.The polymeric films were prepared by compression of the purified resin and annealing the film to induce efficient back formation of the anhydride groups.The properties of the film surface were analyzed by attenuated total reflection Fourier transforms infrared spectroscopy and water contact angle measurements.The protein content was determined by Bradford assay.To obtain optimum conditions,immersion time for protein binding was examined.Results revealed that proteins can be successfully immobilized onto the film surface via covalent linkage.The efficiency of the covalent binding of the extractable protein to maleic anhydride-polyethylene film was estimated at 69.87 μtg/cm2,although the film had low anhydride content (3%) on the surface.

  4. Effects of resistance training and protein supplementation on bone turnover in young adult women

    Directory of Open Access Journals (Sweden)

    Sinning Wayne E

    2005-08-01

    Full Text Available Abstract Background The strength of aging bone depends on the balance between the resorption and formation phases of the remodeling process. The purpose of this study was to examine the interaction of two factors with the potential to exert opposing influences on bone turnover, resistance exercise training and high dietary protein intake. It was hypothesized that resistance training by young, healthy, untrained women with protein intakes near recommended levels (0.8 g·kg-1·d-1 would promote bone formation and/or inhibit bone resorption, and that subsequent supplementation to provide 2.4 g protein·kg-1·d-1 would reverse these effects. Methods Bone formation was assessed with serum bone-specific alkaline phosphatase (BAP and osteocalcin (OC, and bone resorption with urinary calcium and deoxypyridinoline (DPD. Biochemical, strength, anthropometric, dietary, and physical activity data were obtained from 24 healthy, untrained, eumenorrheic women (18–29y at baseline, after eight weeks of resistance training (3 d·wk-1, ~1 hr·d-1; 3 sets, 6–10 repetitions, 13 exercises, 75–85% maximum voluntary contraction, and after 12 weeks of resistance training and 10 days of protein/placebo supplementation. Subjects were randomized (double-blind to either a high protein (HP or training control (TC group and, during the final 10 days, consumed either enough purified whey protein to bring daily protein intake to 2.4 g·kg-1·d-1, or an equivalent dose of isoenergetic, carbohydrate placebo. Results Strength, lean tissue mass, and DPD increased significantly in both groups over time, while percent body fat and BAP decreased (repeated measures ANOVA, p ≤ 0.05, Bonferroni correction. No significant changes were observed for serum OC or urinary calcium, and no significant group (TC, HP × time (baseline, week 8, week 12 interactions emerged for any of the biochemical measures. Conclusion (1 Twelve weeks of high-intensity resistance training did not appear to

  5. Bone Samples Extracted from Embalmed Subjects Are Not Appropriate for the Assessment of Bone Quality at the Molecular Level Using Raman Spectroscopy.

    Science.gov (United States)

    Pascart, Tristan; Cortet, Bernard; Olejnik, Cecile; Paccou, Julien; Migaud, Henri; Cotten, Anne; Delannoy, Yann; During, Alexandrine; Hardouin, Pierre; Penel, Guillaume; Falgayrac, Guillaume

    2016-03-01

    Bone samples extracted from embalmed cadavers are commonly used as controls in the study of bone. The effects of embalmment on the molecular composition of bone are unknown. The objective of this study was to determine the effect of embalmment on the molecular composition and structure of bone, as evaluated by Raman spectroscopy. Bone samples of femoral heads from five embalmed donors and five fresh-frozen donors were compared using Raman microspectroscopy with DuoScan technology. Physicochemical parameters simultaneously describing the organic and mineral phases of bone were compared using the Mann-Whitney U test. Partial least squares discriminant analysis (PLS-DA) was used to determine specific Raman spectral features of each group. Study of the mineral phase showed a 15% reduction of the mineral-to-matrix ratio (p < 0.001), an 8% decrease of type B carbonate substitution (p < 0.001), and a 2% increase in crystallinity (p < 0.001) in the embalmed donors group compared to those of the fresh donors group. Regarding the organic phase of bone, the hydroxyproline-to-proline ratio was increased by 18% in the embalmed group (p < 0.001), with no variation in both the relative proteoglycan content (GAG/CH3) (p = 0.08) and collagen maturity (p = 0.57). PLS-DA showed that the embalmed group was characterized mainly by peaks assigned to hydroxyproline, lipids, and collagen. Embalmment induces significant modifications of the molecular composition of bone. Bone samples from embalmed subjects should be avoided as controls for Raman spectroscopy studies. Preservation procedures performed prior to bone sampling should be reported in studies using human cadaver samples. PMID:26824493

  6. DNA in ancient bone - Where is it located and how should we extract it?

    DEFF Research Database (Denmark)

    Campos, Paula; Craig, Oliver E.; Turner-Walker, Gordon;

    2011-01-01

    . The question arises as to whether this may be due to post-collection preservation or just an artefact of the extraction methods used in these different studies? In an attempt to resolve these questions, we examine the efficacy of DNA extraction methods, and the quality and quantity of DNA recovered......Despite the widespread use of bones in ancient DNA (aDNA) studies, relatively little concrete information exists in regard to how the DNA in mineralised collagen degrades, or where it survives in the material's architecture. While, at the macrostructural level, physical exclusion of microbes and...... other external contaminants may be an important feature, and, at the ultrastructural level, the adsorption of DNA to hydroxyapatite and/or binding of DNA to Type I collagen may stabilise the DNA, the relative contribution of each, and what other factors may be relevant, are unclear. There is...

  7. Extraction site preservation using new graft material that combines mineralized and demineralized allograft bone: a case series report with histology.

    Science.gov (United States)

    Holtzclaw, Dan

    2014-02-01

    This retrospective case series reports on the use of a new allograft bone product composed of a 70 to 30 ratio of mineralized to demineralized cortical bone particles to preserve the alveolar ridge dimensions of patients requiring tooth extraction with plans for future dental implant placement. Ten patients received atraumatic tooth extractions with subsequent placement of the blended bone allograft. All sites were covered with a single layer of amnion-chorion, which was intentionally left exposed. After an average healing time of 14 weeks, the surgical sites were accessed for placement of dental implants. Prior to implant placement, trephine core samples were taken for histologic analysis. Dental implants were successfully placed in all previously grafted surgical sites; no sites required any additional bone augmentation. Histologic analysis of trephined bone core samples revealed bone compositions similar to those described in previously published site preservation studies. The results of this case series suggest that blended bone allograft containing a 70 to 30 ratio of mineralized to demineralized cortical bone particles can be successfully used to facilitate future placement of dental implants with as little as 14 weeks of healing. PMID:24571560

  8. Validation of a Non-Targeted LC-MS Approach for Identifying Ancient Proteins: Method Development on Bone to Improve Artifact Residue Analysis

    Directory of Open Access Journals (Sweden)

    Andrew Barker

    2015-09-01

    Full Text Available Identification of protein residues from prehistoric cooking pottery using mass spectrometry is challenging because proteins are removed from original tissues, are degraded from cooking, may be poorly preserved due to diagenesis, and occur in a palimpsest of exogenous soil proteins. In contrast, bone proteins are abundant and well preserved. This research is part of a larger method-development project for innovation and improvement of liquid chromatography – mass spectrometry analysis of protein residues from cooking pottery; here we validate the potential of our extraction and characterization approach via application to ancient bone proteins. Because of its preservation potential for proteins and given that our approach is destructive, ancient bone identified via skeletal morphology represents an appropriate verification target. Proteins were identified from zooarchaeological turkey (Meleagris gallopavo Linnaeus Phasianidae, rabbit (Lagomorpha, and squirrel (Sciuridae remains excavated from ancient pueblo archaeological sites in southwestern Colorado using a non-targeted LC-MS/MS approach. The data have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD002440. Improvement of highly sensitive targeted LC-MS/MS approaches is an avenue for future method development related to the study of protein residues from artifacts such as stone tools and pottery.

  9. Biorefinery process for protein extraction from oriental mustard (Brassica juncea (L.) Czern.) using ethanol stillage

    OpenAIRE

    Ratanapariyanuch, Kornsulee; Tyler, Robert T.; Shim, Youn Young; Reaney, Martin JT

    2012-01-01

    Large volumes of treated process water are required for protein extraction. Evaporation of this water contributes greatly to the energy consumed in enriching protein products. Thin stillage remaining from ethanol production is available in large volumes and may be suitable for extracting protein rich materials. In this work protein was extracted from ground defatted oriental mustard (Brassica juncea (L.) Czern.) meal using thin stillage. Protein extraction efficiency was studied at pHs betwee...

  10. Bone

    International Nuclear Information System (INIS)

    Bone scanning provides information on the extent of primary bone tumors, on possible metastatic disease, on the presence of osteomyelitis prior to observation of roentgenographic changes so that earlier therapy is possible, on the presence of collagen diseases, on the presence of fractures not disclosed by x-ray films, and on the evaluation of aseptic necrosis. However, the total effect and contribution of bone scanning to the diagnosis, treatment, and ultimate prognosis of pediatric skeletal diseases is, as yet, unknown. (auth)

  11. Osteogenic protein-1 increases the fixation of implants grafted with morcellised bone allograft and ProOsteon bone substitute: an experimental study in dogs

    DEFF Research Database (Denmark)

    Baad-Hansen, Thomas Einer; Overgaard, S; Lind, M;

    2007-01-01

    weeks osteogenic protein-1 increased bone formation and the energy absorption of implants grafted with allograft and ProOsteon. A composite of allograft, ProOsteon and osteogenic protein-1 was comparable, but not superior to, allograft used on its own. ProOsteon alone cannot be recommended as a......Impacted bone allograft is often used in revision joint replacement. Hydroxyapatite granules have been suggested as a substitute or to enhance morcellised bone allograft. We hypothesised that adding osteogenic protein-1 to a composite of bone allograft and non-resorbable hydroxyapatite granules...... surrounded by a concentric 3 mm gap. These gaps were randomly allocated to four different procedures in each dog: 1) bone allograft used on its own; 2) ProOsteon used on its own; 3) allograft and ProOsteon used together; or 4) allograft and ProOsteon with the addition of osteogenic protein-1. After three...

  12. DNA extraction: an anthropologic aspect of bone remains from sixth- to seventh-century ad bone remains.

    Science.gov (United States)

    Di Nunno, Nunzio; Saponetti, Sandro Sublimi; Scattarella, Vito; Emanuel, Patrizia; Baldassarra, Stefania Lonero; Volpe, Giuliano; Di Nunno, Cosimo

    2007-12-01

    In the archeological site of the early Christian Episcopal complex of Saint Peter, in Canosa di Puglia (Bari, Italy), during the operations of archaeological excavations, tombs were discovered. They were dated between the sixth and seventh centuries ad with carbon 14 methodology. Five skeletons were found in the 5 tombs: 28A: male individual, 43 years old. The height was 170 cm; the biomass was 65.7 kg. The analysis of the bones indicated several noteworthy pathologies, such as a number of hypoplasia lines of the enamel, the presence of Schmorl hernias on the first 2 lumbar vertebrae, and the outcome of subacromial impingement syndrome. 28E was a male individual, with a biologic age of death of between 44 and 60 years. The height was 177 cm. He had a posttraumatic fracture callus of the medial third of the clavicle, with an oblique fracture rima. 29B was a female individual, 44-49 years old. The height was 158.8 cm; the biomass was 64.8 kg. There was Wells bursitis on the ischial tuberosity on both sides. 29E was a male individual, 45-50 years old. The height was 169.47 cm; the biomass was 70.8 kg. The third and the fourth vertebrae showed Baastrup syndrome (compression of the vertebral spine). There were radiologic signs of deformity on the higher edge of the acetabula and results of frequent sprains of the ankles. 31A was a male individual, 47-54 years old. The height was 178.65 cm; the biomass was 81 kg. The vertebral index showed a heavy overloading in the thoracic lumbar region. There were bony formations under the periosteum on both on the higher and medium facets of the first metatarsus and on the higher and lateral facets of the fifth metatarsus on both sides. As the topography indicates, these small ossifications coincided with the contact points between the back of the foot and parts of the upper shoe. From the osseous remains, in particular from the teeth (central incisors), the DNA was extracted and typed to identify potential family ties among all the

  13. GPCR kinase 2 interacting protein 1 (GIT1) regulates osteoclast function and bone mass

    OpenAIRE

    Menon, Prashanthi; Yin, Guoyong; Smolock, Elaine M.; Zuscik, Michael J.; Yan, Chen; Berk, Bradford C.

    2010-01-01

    G-protein coupled receptor (GPCR) kinase 2 interacting protein-1 (GIT1) is a scaffold protein expressed in various cell types including neurons, endothelial and vascular smooth muscle cells. The GIT1 knockout (KO) mouse has a pulmonary phenotype due to impaired endothelial function. Because GIT1 is tyrosine phosphorylated by Src kinase, we anticipated that GIT1 KO should have a bone phenotype similar to Src KO. Microcomputed tomography of the long bones revealed that GIT1 KO mice have a 2.3-f...

  14. Osteogenic protein 1 device increases bone formation and bone graft resorption around cementless implants

    DEFF Research Database (Denmark)

    Jensen, Thomas B; Overgaard, Søren; Lind, Martin;

    2002-01-01

    In each femoral condyle of 8 Labrador dogs, a non weight-bearing hydroxyapatite-coated implant was inserted surrounded by a 3 mm gap. Each gap was filled with bone allograft or ProOsteon with or without OP-1 delivered in a bovine collagen type I carrier (OP-1 device). 300 microg OP-1 was used in ...

  15. Enzymatic aqueous technology for simultaneous coconut protein and oil extraction

    OpenAIRE

    Coelho, Maria A. Z.; Suely P. Freitas; Sant’Anna, Beatriz P.M.

    2003-01-01

    In this study, the following commercial enzymes were evaluated in the enzymatic extraction of oil and protein from coconut: Celluclast, Termamyl, Viscozyme, Neutrase y Protease. Preliminary experiments were carried out for selection of enzymes, enzyme concentration and incubation time. In order to verify the effects of pH and water/substrate ratio, a response surface methodology was applied based in a 32 full factorial experimental design. The pH is the most meaningful parameter on oil and pr...

  16. Automatically extracting functionally equivalent proteins from SwissProt

    Directory of Open Access Journals (Sweden)

    Martin Andrew CR

    2008-10-01

    Full Text Available Abstract Background There is a frequent need to obtain sets of functionally equivalent homologous proteins (FEPs from different species. While it is usually the case that orthology implies functional equivalence, this is not always true; therefore datasets of orthologous proteins are not appropriate. The information relevant to extracting FEPs is contained in databanks such as UniProtKB/Swiss-Prot and a manual analysis of these data allow FEPs to be extracted on a one-off basis. However there has been no resource allowing the easy, automatic extraction of groups of FEPs – for example, all instances of protein C. We have developed FOSTA, an automatically generated database of FEPs annotated as having the same function in UniProtKB/Swiss-Prot which can be used for large-scale analysis. The method builds a candidate list of homologues and filters out functionally diverged proteins on the basis of functional annotations using a simple text mining approach. Results Large scale evaluation of our FEP extraction method is difficult as there is no gold-standard dataset against which the method can be benchmarked. However, a manual analysis of five protein families confirmed a high level of performance. A more extensive comparison with two manually verified functional equivalence datasets also demonstrated very good performance. Conclusion In summary, FOSTA provides an automated analysis of annotations in UniProtKB/Swiss-Prot to enable groups of proteins already annotated as functionally equivalent, to be extracted. Our results demonstrate that the vast majority of UniProtKB/Swiss-Prot functional annotations are of high quality, and that FOSTA can interpret annotations successfully. Where FOSTA is not successful, we are able to highlight inconsistencies in UniProtKB/Swiss-Prot annotation. Most of these would have presented equal difficulties for manual interpretation of annotations. We discuss limitations and possible future extensions to FOSTA, and

  17. The retinoblastoma protein tumor suppressor is important for appropriate osteoblast differentiation and bone development

    OpenAIRE

    Berman, Seth D.; Yuan, Tina L.; Miller, Emily S.; Lee, Eunice; Caron, Alicia; Lees, Jacqueline A

    2008-01-01

    Mutation of the Rb tumor suppressor gene is strongly linked to osteosarcoma formation. This observation, and the documented interaction between the retinoblastoma protein (pRb) and Runx2, suggests that pRb is important in bone development. To assess this hypothesis, we used a conditional knockout strategy to generate pRb-deficient embryos that survive to birth. Analysis of these embryos shows that Rb-inactivation causes the abnormal development and impaired ossification of several bones, corr...

  18. Effects of dietary protein and glycaemic index on biomarkers of bone turnover in children

    DEFF Research Database (Denmark)

    Dalskov, Stine-Mathilde; Müller, Martha; Ritz, Christian;

    2014-01-01

    For decades, it has been debated whether high protein intake compromises bone mineralisation, but no long-term randomised trial has investigated this in children. In the family-based, randomised controlled trial DiOGenes (Diet, Obesity and Genes), we examined the effects of dietary protein and...... glycaemic index (GI) on biomarkers of bone turnover and height in children aged 5-18 years. In two study centres, families with overweight parents were randomly assigned to one of five ad libitum-energy, low-fat (25-30 % energy (E%)) diets for 6 months: low protein/low GI; low protein/high GI; high protein....../low GI; high protein/high GI; control. They received dietary instructions and were provided all foods for free. Children, who were eligible and willing to participate, were included in the study. In the present analyses, we included children with data on plasma osteocalcin or urinary N...

  19. Demineralized dentin matrix combined with recombinant human bone morphogenetic protein-2 in rabbit calvarial defects

    Science.gov (United States)

    2016-01-01

    Objectives The aim of this study was to compare the osteogenic effects of demineralized dentin matrix (DDM) combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) in rabbit calvarial defects with DDM and anorganic bovine bone (ABB) combined with rhBMP-2. Materials and Methods Four round defects with 8-mm diameters were created in each rabbit calvaria. Each defect was treated with one of the following: 1) DDM, 2) ABB/rhBMP-2, or 3) DDM/rhBMP-2. The rhBMP-2 was combined with DDM and ABB according to a stepwise dry and dip lyophilizing protocol. Histological and microcomputed tomography (µCT) analyses were performed to measure the amount of bone formation and bone volume after 2- and 8-week healing intervals. Results Upon histological observation at two weeks, the DDM and ABB/rhBMP-2 groups showed osteoconductive bone formation, while the DDM/rhBMP-2 group showed osteoconductive and osteoinductive bone formation. New bone formation was higher in DDM/rhBMP-2, DDM and ABB decreasing order. The amounts of bone formation were very similar at two weeks; however, at eight weeks, the DDM/rhBMP-2 group showed a two-fold greater amount of bone formation compared to the DDM and ABB/rhBMP-2 groups. The µCT analysis showed markedly increased bone volume in the DDM/rhBMP-2 group at eight weeks compared with that of the DDM group. Notably, there was a slight decrease in bone volume in the ABB/rhBMP-2 group at eight weeks. There were no significant differences among the DDM, ABB/rhBMP-2, and DDM/rhBMP-2 groups at two or eight weeks. Conclusion Within the limitations of this study, DDM appears to be a suitable carrier for rhBMP-2 in orthotopic sites. PMID:27162749

  20. Extraction and downstream processing of plant-derived recombinant proteins.

    Science.gov (United States)

    Buyel, J F; Twyman, R M; Fischer, R

    2015-11-01

    Plants offer the tantalizing prospect of low-cost automated manufacturing processes for biopharmaceutical proteins, but several challenges must be addressed before such goals are realized and the most significant hurdles are found during downstream processing (DSP). In contrast to the standardized microbial and mammalian cell platforms embraced by the biopharmaceutical industry, there are many different plant-based expression systems vying for attention, and those with the greatest potential to provide inexpensive biopharmaceuticals are also the ones with the most significant drawbacks in terms of DSP. This is because the most scalable plant systems are based on the expression of intracellular proteins in whole plants. The plant tissue must therefore be disrupted to extract the product, challenging the initial DSP steps with an unusually high load of both particulate and soluble contaminants. DSP platform technologies can accelerate and simplify process development, including centrifugation, filtration, flocculation, and integrated methods that combine solid-liquid separation, purification and concentration, such as aqueous two-phase separation systems. Protein tags can also facilitate these DSP steps, but they are difficult to transfer to a commercial environment and more generic, flexible and scalable strategies to separate target and host cell proteins are preferable, such as membrane technologies and heat/pH precipitation. In this context, clarified plant extracts behave similarly to the feed stream from microbes or mammalian cells and the corresponding purification methods can be applied, as long as they are adapted for plant-specific soluble contaminants such as the superabundant protein RuBisCO. Plant-derived pharmaceutical proteins cannot yet compete directly with established platforms but they are beginning to penetrate niche markets that allow the beneficial properties of plants to be exploited, such as the ability to produce 'biobetters' with tailored

  1. CCAAT/enhancer binding protein β-deficiency enhances type 1 diabetic bone phenotype by increasing marrow adiposity and bone resorption

    OpenAIRE

    Motyl, Katherine J.; Raetz, Michelle; Tekalur, Srinivasan Arjun; Schwartz, Richard C.; McCabe, Laura R.

    2011-01-01

    Bone loss in type 1 diabetes is accompanied by increased marrow fat, which could directly reduce osteoblast activity or result from altered bone marrow mesenchymal cell lineage selection (adipocyte vs. osteoblast). CCAAT/enhancer binding protein beta (C/EBPβ) is an important regulator of both adipocyte and osteoblast differentiation. C/EBPβ-null mice have delayed bone formation and defective lipid accumulation in brown adipose tissue. To examine the balance of C/EBPβ functions in the diabetic...

  2. A Novel HA/β-TCP-Collagen Composite Enhanced New Bone Formation for Dental Extraction Socket Preservation in Beagle Dogs

    Directory of Open Access Journals (Sweden)

    Ko-Ning Ho

    2016-03-01

    Full Text Available Past studies in humans have demonstrated horizontal and vertical bone loss after six months following tooth extraction. Many biomaterials have been developed to preserve bone volume after tooth extraction. Type I collagen serves as an excellent delivery system for growth factors and promotes angiogenesis. Calcium phosphate ceramics have also been investigated because their mineral chemistry resembles human bone. The aim of this study was to compare the performance of a novel bioresorbable purified fibrillar collagen and hydroxyapatite/β-tricalcium phosphate (HA/β-TCP ceramic composite versus collagen alone and a bovine xenograft-collagen composite in beagles. Collagen plugs, bovine graft-collagen composite and HA/β-TCP-collagen composite were implanted into the left and right first, second and third mandibular premolars, and the fourth molar was left empty for natural healing. In total, 20 male beagle dogs were used, and quantitative and histological analyses of the extraction ridge was done. The smallest width reduction was 19.09% ± 8.81% with the HA/β-TCP-collagen composite at Week 8, accompanied by new bone formation at Weeks 4 and 8. The HA/β-TCP-collagen composite performed well, as a new osteoconductive and biomimetic composite biomaterial, for socket bone preservation after tooth extraction.

  3. Depot injectable biodegradable nanoparticles loaded with recombinant human bone morphogenetic protein-2: preparation, characterization, and in vivo evaluation

    Directory of Open Access Journals (Sweden)

    Hassan AH

    2015-07-01

    Full Text Available Ali Habiballah Hassan,1 Khaled Mohamed Hosny,2,3 Zuahir A Murshid,1 Adel Alhadlaq,4 Ahmed Alyamani,5 Ghada Naguib6 1Department of Orthodontics, Faculty of Dentistry, 2Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Saudi Arabia; 3Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmacy, Beni Suef University, Beni Suef, Egypt; 4Department of Pediatric Dentistry and Orthodontics, College of Dentistry, King Saud University, Riyadh, 5Department of Oral Surgery, 6Department of Restorative Dentistry, Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia Objective: The aim of this study is to utilize the biocompatibility characteristics of biodegradable polymers, viz, poly lactide-co-glycolide (PLGA and polycaprolactone (PCL, to prepare sustained-release injectable nanoparticles (NPs of bone morphogenetic protein-2 (BMP-2 for the repair of alveolar bone defects in rabbits. The influence of formulation parameters on the functional characteristics of the prepared NPs was studied to develop a new noninvasive injectable recombinant human BMP-2 (rhBMP-2 containing grafting material for the repair of alveolar bone clefts.Materials and methods: BMP-2 NPs were prepared using a water-in-oil-in-water double-emulsion solvent evaporation/extraction method. The influence of molar ratio of PLGA to PCL on a suitable particle size, encapsulation efficiency, and sustained drug release was studied. Critical size alveolar defects were created in the maxilla of 24 New Zealand rabbits divided into three groups, one of them treated with 5 µg/kg of rhBMP-2 NP formulations.Results: The results found that NPs formula prepared using blend of PLGA and PCL in 4:2 (w/w ratio showed the best sustained-release pattern with lower initial burst, and showed up to 62.7% yield, 64.5% encapsulation efficiency, 127 nm size, and more than 90% in vitro release. So, this formula was selected for

  4. Intramembranous bone healing process subsequent to tooth extraction in mice: micro-computed tomography, histomorphometric and molecular characterization.

    Science.gov (United States)

    Vieira, Andreia Espindola; Repeke, Carlos Eduardo; Ferreira Junior, Samuel de Barros; Colavite, Priscila Maria; Biguetti, Claudia Cristina; Oliveira, Rodrigo Cardoso; Assis, Gerson Francisco; Taga, Rumio; Trombone, Ana Paula Favaro; Garlet, Gustavo Pompermaier

    2015-01-01

    Bone tissue has a significant potential for healing, which involves a significant the interplay between bone and immune cells. While fracture healing represents a useful model to investigate endochondral bone healing, intramembranous bone healing models are yet to be developed and characterized. In this study, a micro-computed tomography, histomorphometric and molecular (RealTimePCRarray) characterization of post tooth-extraction alveolar bone healing was performed on C57Bl/6 WT mice. After the initial clot dominance (0 h), the development of a provisional immature granulation tissue is evident (7 d), characterized by marked cell proliferation, angiogenesis and inflammatory cells infiltration; associated with peaks of growth factors (BMP-2-4-7,TGFβ1,VEGFa), cytokines (TNFα, IL-10), chemokines & receptors (CXCL12, CCL25, CCR5, CXCR4), matrix (Col1a1-2, ITGA4, VTN, MMP1a) and MSCs (CD105, CD106, OCT4, NANOG, CD34, CD146) markers expression. Granulation tissue is sequentially replaced by more mature connective tissue (14 d), characterized by inflammatory infiltrate reduction along the increased bone formation, marked expression of matrix remodeling enzymes (MMP-2-9), bone formation/maturation (RUNX2, ALP, DMP1, PHEX, SOST) markers, and chemokines & receptors associated with healing (CCL2, CCL17, CCR2). No evidences of cartilage cells or tissue were observed, strengthening the intramembranous nature of bone healing. Bone microarchitecture analysis supports the evolving healing, with total tissue and bone volumes as trabecular number and thickness showing a progressive increase over time. The extraction socket healing process is considered complete (21 d) when the dental socket is filled by trabeculae bone with well-defined medullary canals; it being the expression of mature bone markers prevalent at this period. Our data confirms the intramembranous bone healing nature of the model used, revealing parallels between the gene expression profile and the

  5. An Improvement on the Protein Extraction Procedure from Eggshell Membrane

    Institute of Scientific and Technical Information of China (English)

    Peng Ruo-ting; Guo Yulong

    2010-01-01

    @@ 1 Introduction Collagen is a frequently occurring protein found in poultry,livestock and human body,esp.in the hair,skin,bone cartilage,tendon and so on.It forms the strongly insoluble fibers and serves as connective tissue supporting materials.Nowadays it became one of the most important raw materials of bio-engineering materials which are widely used in various fields including cosmetics,biomedical and foods industries.With the outburst of multiple diseases from cows and pigs such as bovine spongiform encephalopathy(BSE),related authorities in different countries have estabhshed many restrictions on trading collagen from mammalians.Due to this ground scientists began to identify new resources of collagens.

  6. Recombinant human bone morphogenetic protein-7 expressed from CHO cells possessing the activity of bone-induced in vitro

    Institute of Scientific and Technical Information of China (English)

    LI Xiaoyan; WANG Hao; YANG Yang; TAN Min; XUE Jingya; NI Haidong; GUO Yajun

    2006-01-01

    Objective To express the recombinant human bone morphogenetic protein-7 (rhBMP-7) in Chinese hamster ovary (CHO) cells and to establish the in vitro biological activity assay of rhBMP-7. Methods Human BMP-7 cDNA was subcloned into pcDNA3.1 mammalian expression vector and transfected to CHO cells by using the lipofectin transfection method. BMP-7 expression cell culture supernatants were harvested and purified for target protein. To analyze the bioactivity of the secreted rhBMP-7, a novel in vitro assay was established by measuring its alkaline phosphatase (ALP) stimulating of osteoblast cell line, W-20-17. Results BMP-7 stably expressing cell clone was selected, which secreted mature disulfide-linked homodimer form of hBMP-7 and had an apparent molecular weight of 36kDa. rhBMP-7 with >95% purity was obtained using 3 step chromatography method. Bioactivity assay showed that the purified protein specifically stimulated W-20-17 cell producing ALP, with a 4-fold increase of ALP activity at 100ng/ml or more, and the EC50 of 15.6ng/ml. Conclusion Purified rhBMP-7 from this CHO expression system has significant biological activity in induction of osteoblast phenotype, which demonstrates potential bone regeneration activity.

  7. The Inhibitory Effects of Forsythia Koreana Extracts on the Metastatic Ability of Breast Cancer Cells and Bone Resorption by Osteoclasts

    Science.gov (United States)

    Kim, Yu Li; Lee, Sun Kyoung; Park, Kwang-Kyun; Chung, Won-Yoon

    2016-01-01

    Background: Breast cancer is the most common malignant disease in women. The patients with advanced breast cancer develop metastasis to bone. Bone metastasis and skeletal-related events by breast cancer are frequently associated with the invasiveness of breast cancer cells and osteoclasts-mediated bone resorption. Forsythia koreana is used in oriental traditional medicine to treat asthma, atopy, and allergic diseases. The aim of this study was to evaluate the inhibitory effects of F. koreana extracts on the invasion of breast cancer cells and bone resorption by osteoclasts. Methods: Cell viability was measured by an MTT assay and the migration and invasion of MDA-MB-231 cells were detected by a Boyden chamber assay. The formation of osteoclasts and pit was detected using tartrate-resistant acid phosphatase staining and calcium phosphate-coated plates, respectively. The activities of matrix metalloproteinases (MMPs) and cathepsin K were evaluated by gelatin zymography and a cathepsin K detection kit. Results: The fruit and leaf extracts of F. koreana significantly inhibited the invasion of MDA-MB-231 cells at noncytotoxic concentrations. The fruit extract of F. koreana reduced the transforming growth factor β1-induced migration, invasion and MMPs activities of MDA-MB-231 cells. In addition, the fruit, branch, and leaf extracts of F. koreana also inhibited the receptor activator of nuclear factor kappa-B ligand-induced osteoclast formation and osteoclast-mediated bone-resorbing activity by reducing the activities of MMPs and cathepsin K. Conclusions: The extracts of F. koreana may possess the potential to inhibit the breast cancer-induced bone destruction through blocking invasion of breast cancer cells, osteoclastogenesis, and the activity of mature osteoclasts. PMID:27390737

  8. Reverse micellar extraction for downstream processing of proteins/enzymes.

    Science.gov (United States)

    Krishna, S Hari; Srinivas, N D; Raghavarao, K S M S; Karanth, N G

    2002-01-01

    New developments in the area of downstream processing are, hopefully, to fulfill the promises of modern biotechnology. The traditional separation processes such as chromatography or electrophoresis can become prohibitively expensive unless the product is of high value. Hence, there is a need to develop efficient and cost-effective downstream processing methods. Reverse micellar extraction is one such potential and a promising liquid-liquid extraction technique, which has received immense attention for isolation and purification of proteins/enzymes in the recent times. This technique is easy to scale-up and offers continuous operation. This review, besides briefly considering important physico-chemical and biological aspects, highlights the engineering aspects including mass transfer, mathematical modeling, and technology development. It also discusses recent developments in reverse micellar extraction such as affinity based separations, enzymatic reactions in reverse micelles coupled with membrane processes, reverse micellar extraction in hollow fibers, etc. Special emphasis has been given to some recent applications of this technique. PMID:11787493

  9. Methodological approach for the evaluation of homologous bone graft use in post-extractive atrophic alveolar ridges

    Directory of Open Access Journals (Sweden)

    B. Musante

    2013-01-01

    Full Text Available The aim of this study is to investigate the in vivo efficacy of Puros® cancellous particulate allograft bone in the regeneration of post-extractive sites. Twelve molar or premolar sites were selected. Seven days after minimally invasive tooth extraction, Puros® cancellous particulate allografts were inserted into the elected sites. TC Cone-Beam investigation and at 4 months from surgery; bone height and width were evaluated. Five months after surgery, biopsy samples of the regenerated sites were taken and histologically analyzed to qualitatively evaluate bone regeneration. TC analysis showed, a mean bone gain of 4.1 mm in height and 2.02 mm in width. The histological analysis of the samples showed intense bone metabolic activity with active osteoblasts on the implant surface, at the level of the native bone-graft interface and in the grafted area. The radiographic and histological analyses demonstrate an optimal bonen regeneration, both in terms of quality and quantity using Puros®.

  10. Effect of the "protein diet" and bone tissue

    OpenAIRE

    Zoraide Nascimento da Silva; Vanessa Azevedo de Jesuz; Eduardo de Salvo Castro; Carlos Alberto Soares da Costa; Gilson Teles Boaventura; Vilma Blondet de Azeredo

    2014-01-01

    The aim of this study is to evaluate the effect of the hyperproteic diet consumption on bone tissue. Methods: The study was conducted during sixty days. Twenty eight Wistar albinus rats, adults, originated from Laboratory of Experimental Nutrition were divided in four groups: (n = 7); Control 1 (C1), Control 2 (C2), Hyperproteic 1 (HP1) e Hyperproteic 2 (HP2). The C2 and HP2 groups were submitted to 30% of food restriction. The hyperproteic diet was based on the Atkins diet and prepared to si...

  11. Comparison of a quasi-dynamic and a static extraction method for the cytotoxic evaluation of acrylic bone cements.

    Science.gov (United States)

    Hoess, A; López, A; Engqvist, H; Ott, M Karlsson; Persson, C

    2016-05-01

    In this study, two different extraction approaches were compared in order to evaluate the cytotoxicity of 7 different acrylic bone cements, mainly developed for spinal applications, to osteoblastic cells. Firstly, a static extraction was carried out continuously over 24h, a method widely used in literature. Secondly, a quasi-dynamic extraction method that allowed the investigation of time-dependent cytotoxic effects of curing acrylic bone cements to cells was introduced. In both cases the extraction of the cements was started at a very early stage of the polymerization process to simulate the conditions during clinical application. Data obtained by the quasi-dynamic extraction method suggest that the cytotoxicity of the setting materials mainly originates from the release of toxic components during the first hour of the polymerization reaction. It was also shown that a static extraction over 24h generally represents this initial stage of the curing process. Furthermore, compared to the static extraction, time-dependent cytotoxicity profiles could be detected using the quasi-dynamic extraction method. Specifically, a modification of commercial OsteopalV with castor oil as a plasticizer as well as a customized cement formulation showed clear differences in cytotoxic behavior compared to the other materials during the setting process. In addition, it was observed that unreacted monomer released from the castor oil modified cement was not the main component affecting the toxicity of the material extracts. The quasi-dynamic extraction method is a useful tool to get deeper insight into the cytotoxic potential of curing acrylic bone cements under relevant biological conditions, allowing systematic optimization of materials under development. PMID:26952424

  12. Mechanical microenvironments and protein expression associated with formation of different skeletal tissues during bone healing.

    Science.gov (United States)

    Miller, Gregory J; Gerstenfeld, Louis C; Morgan, Elise F

    2015-11-01

    Uncovering the mechanisms of the sensitivity of bone healing to mechanical factors is critical for understanding the basic biology and mechanobiology of the skeleton, as well as for enhancing clinical treatment of bone injuries. This study refined an experimental method of measuring the strain microenvironment at the site of a bone injury during bone healing. This method used a rat model in which a well-controlled bending motion was applied to an osteotomy to induce the formation of pseudarthrosis that is composed of a range of skeletal tissues, including woven bone, cartilage, fibrocartilage, fibrous tissue, and clot tissue. The goal of this study was to identify both the features of the strain microenvironment associated with formation of these different tissues and the expression of proteins frequently implicated in sensing and transducing mechanical cues. By pairing the strain measurements with histological analyses that identified the regions in which each tissue type formed, we found that formation of the different tissue types occurs in distinct strain microenvironments and that the type of tissue formed is correlated most strongly to the local magnitudes of extensional and shear strains. Weaker correlations were found for dilatation. Immunohistochemical analyses of focal adhesion kinase and rho family proteins RhoA and CDC42 revealed differences within the cartilaginous tissues in the calluses from the pseudarthrosis model as compared to fracture calluses undergoing normal endochondral bone repair. These findings suggest the involvement of these proteins in the way by which mechanical stimuli modulate the process of cartilage formation during bone healing. PMID:25822264

  13. Osseointegration of titanium implants by addition of recombinant bone morphogenetic protein 2 (rhBMP-2)

    Energy Technology Data Exchange (ETDEWEB)

    Lichtinger, T.K.; Mueller, R.T.; Schuermann, N.; Oldenburg, M. [Essen Univ. (Germany). Dept. of Orthopaedic Surgery; Wiemann, M. [Inst. of Physiology, Univ. of Essen (Germany); Chatzinikolaidou, M.; Jennissen, H.P. [Inst. of Physiological Chemistry, Univ. of Essen (Germany); Rumpf, H.M.

    2001-12-01

    The osseointegration of long-term implants is often incomplete such that gaps remain between the implant surface and the surrounding hard tissue. This study examines the effect of soluble recombinant human bone morphogenic protein 2 (rhBMP-2) on gap healing and osseous integration. The effect of a single, intraoperative application of soluble rhBMP-2 on the formation of new bone around titanium implants was studied. A total of 8 titanium-alloy cylinders (Ti-6Al-4V) with a plasma spray coating (TPS; 400 {mu}m thickness) were implanted into femoral condyles of mature sheep: rhBMP-2 solution (1 {mu}g) was pipetted into the 1 mm wide cleft around 4 implants; 4 further implants served as rhBMP-2-free controls. Two of these controls exhibited an additional calciumphosphate-coating. The cleft around the implants served as testing zone to study the formation of new bone by microradiographical and histological analyses. The follow-up periods were 4 and 9 weeks, respectively. A significant amount of new bone contacting the implants' surface was detected where rhBMP-2-solution had been used: In 50% a circumferential osseoinduction occurred within 4 weeks and a nearly complete osseointegration was observed after 9 weeks. In all cases bone formation was exaggerated and filled the spongiosa with compact bone. Time matched TPS-controls and controls with calciumphosphate coating showed no notable formation of new bone. The results suggest that a single administration of soluble rhBMP-2 into a bone cavity can augment bone formation and also osseointegration of titanium implants. Further investigations based on these findings are necessary to develop long-term implants (e.g. joint replacements) with rhBMP-2-biocoating for humans. (orig.)

  14. A Shortest Dependency Path Based Convolutional Neural Network for Protein-Protein Relation Extraction

    Science.gov (United States)

    Quan, Chanqin

    2016-01-01

    The state-of-the-art methods for protein-protein interaction (PPI) extraction are primarily based on kernel methods, and their performances strongly depend on the handcraft features. In this paper, we tackle PPI extraction by using convolutional neural networks (CNN) and propose a shortest dependency path based CNN (sdpCNN) model. The proposed method (1) only takes the sdp and word embedding as input and (2) could avoid bias from feature selection by using CNN. We performed experiments on standard Aimed and BioInfer datasets, and the experimental results demonstrated that our approach outperformed state-of-the-art kernel based methods. In particular, by tracking the sdpCNN model, we find that sdpCNN could extract key features automatically and it is verified that pretrained word embedding is crucial in PPI task. PMID:27493967

  15. A randomized controlled evaluation of alveolar ridge preservation following tooth extraction using deproteinized bovine bone mineral and demineralized freeze-dried bone allograft

    Science.gov (United States)

    Sadeghi, Rokhsareh; Babaei, Maryam; Miremadi, S. Asghar; Abbas, Fatemeh Mashadi

    2016-01-01

    Background: Alveolar ridge preservation could be performed immediately following tooth extraction to limit dimensional changes of alveolar process due to bone resorption. The aim of this study was to compare the clinical and histologic outcomes of socket preservation using two different graft materials; deproteinized bovine bone mineral (DBBM) and demineralized freeze-dried bone allograft (DFDBA) with absorbable collagen membrane. Materials and Methods: Twenty extraction sockets in 20 patients were randomly divided into 2 treatment groups: 10 sockets were augmented with DBBM and collagen membrane whereas 10 sockets were filled with DFDBA and covered by collagen membrane. Primary closure was achieved over extraction sockets by flap advancement. Horizontal and vertical ridge dimensional changes were assessed at baseline and after 4-6 months at the time of implant placement. For histological and histomorphometrical analysis, bone samples were harvested from the augmented sites with trephine during implant surgery. All data were analyzed using SPSS version 18 (α=0.05). Results: Clinical measurements revealed that average horizontal reduction was 2.3 ± 0.64 mm for DFDBA and 2.26 ± 0.51 mm for DBBM. Mean vertical ridge resorption at buccal side was 1.29 ± 0.68 mm for DFDBA and 1.1 ± 0.17 mm for DBBM. Moreover, mean vertical ridge reduction at lingual site was 0.41 ± 0.38 mm and 0.35 ± 0.34 mm for DFDBA and DBBM, respectively. No significant differences were seen between two groups in any of those clinical parameters. Histologic analysis showed statistically significant more new bone deposition for DFDBA compared to DBBM (34.49 ± 3.19 vs. 18.76 ± 3.54) (P < 0.01). Residual graft particles were identified significantly more in DBBM (12.77 ± 1.85) than DFDBA (6.06 ± 1.02). Conclusion: Based on the findings of this study, both materials have positive effect on alveolar ridge preservation after tooth extraction, but there was more new bone formation and less

  16. Experimental pharmacological investigation of the antiarthrotic effects of the cartilage and bone marrow extract Rumalon

    Energy Technology Data Exchange (ETDEWEB)

    Kalbhen, D.A.

    1981-08-05

    On the basis of animal experiments, the authors have developed a model of arthrosis which is compatible in its radiological, macroscopic, biochemical, and histological aspects with the pathophysiology of human arthrosis and has been tried in the testing of the antiarthrotic properties of pharmaceuticals. Biochemically induced gonarthroses of experimental animals were used for studies of the effects of a cartilage and bone marrow extract (Rumalon) and a cartilage extract and its high-molecular component DAK-16 on the frequency and progression of degenerative joint diseases. As test parameters, measurements of the articular space, X-ray findings, and macroscopic findings were quantitatively evaluated. The animal experiments show that the inhibitive effects of steroidal and nonsteroidal antirheumatics on the synthesis of the cartilage matrix can be prevented or reduced by simultaneous administration of chondroprotective pharmaceuticals; this may be important on the clinical sector. This antagonism between antiphlogistic agents and Rumalon, which has been observed also in fibroblast cultures and wound healing experiments, is of interest especially for the treatment of activated arthroses.

  17. Experimental pharmacological investigation of the antiarthrotic effects of the cartilage and bone marrow extract Rumalon

    International Nuclear Information System (INIS)

    On the basis of animal experiments, the authors have developed a model of arthrosis which is compatible in its radiological, macroscopic, biochemical, and histological aspects with the pathophysiology of human arthrosis and has been tried in the testing of the antiarthrotic properties of pharmaceuticals. Biochemically induced gonarthroses of experimental animals were used for studies of the effects of a cartilage and bone marrow extract (Rumalon) and a cartilage extract and its high-molecular component DAK-16 on the frequency and progression of degenerative joint diseases. As test parameters, measurements of the articular space, X-ray findings, and macroscopic findings were quantitatively evaluated. The animal experiments show that the inhibitive effects of steroidal and nonsteroidal antirheumatics on the synthesis of the cartilage matrix can be prevented or reduced by simultaneous administration of chondroprotective pharmaceuticals; this may be important on the clinical sector. This antagonism between antiphlogistic agents and Rumalon, which has been observed also in fibroblast cultures and wound healing experiments, is of interest especially for the treatment of activated arthroses. (orig./MG)

  18. The CCN Family Proteins: Modulators of Bone Development and Novel Targets in Bone-Associated Tumors

    OpenAIRE

    Po-Chun Chen; Hsu-Chen Cheng; Shun-Fa Yang; Chiao-Wen Lin; Chih-Hsin Tang

    2014-01-01

    The CCN family of proteins is composed of six extracellular matrix-associated proteins that play crucial roles in skeletal development, wound healing, fibrosis, and cancer. Members of the CCN family share four conserved cysteine-rich modular domains that trigger signal transduction in cell adhesion, migration, proliferation, differentiation, and survival through direct binding to specific integrin receptors and heparan sulfate proteoglycans. In the present review, we discuss the roles of the ...

  19. Bone morphogenetic protein-2 is a negative regulator of hepatocyte proliferation downregulated in the regenerating liver

    NARCIS (Netherlands)

    Xu, Cui-Ping; Ji, Wen-Min; van den Brink, Gijs R.; Peppelenbosch, Maikel P.

    2006-01-01

    AIM: To characterize the expression and dynamic changes of bone morphogenetic protein (BMP)-2 in hepatocytes in the regenerating liver in rats after partial hepatectomy (PH), and examine the effects of BMP-2 on proliferation of human Huh7 hepatoma cells. METHODS: Fifty-four adult male Wistar rats we

  20. Associations of total, dairy, and meat protein with markers for bone turnover in healthy, prepubertal boys

    DEFF Research Database (Denmark)

    Budek, Alicja Zofia; Hoppe, Camilla; Michaelsen, Kim Fleischer;

    2007-01-01

    We previously reported that high intake of milk, but not meat, equal in protein content, increased serum insulin-like growth factor-I (sIGF-I) in prepubertal boys. sIGF-I plays a key role in bone metabolism. Therefore, the aim of this cross-sectional study was to investigate associations of total......, dairy, and meat protein intake with markers for bone turnover and sIGF-I in prepubertal, healthy boys (n ¼ 81). We measured bone turnover (enzyme-linked immunoassay) in serum osteocalcin (sOC), bone-specific alkaline phosphatase (sBAP), and C-terminal telopeptide of collagen type-I (sCTX); dietary...... intake was estimated from a 3-d weighed food record. sIGF-I and its binding protein-3 were assessed (immunoassay) in a subgroup of 56 boys. All statistical models included effects of age, BMI, and energy intake. Dairy protein was negatively associated with sOC (P ¼ 0.05) but not significantly associated...

  1. The expression of bone morphogenetic proteins in osteosarcoma and its relevance as a prognostic parameter

    OpenAIRE

    Sulzbacher, I; Birner, P; Trieb, K; Pichlbauer, E; Lang, S.

    2002-01-01

    Aims: The expression of bone morphogenetic proteins (BMPs) was analysed in 47 osteosarcomas to determine differences in the expression of BMP subtypes and to correlate expression with response to chemotherapy, in addition to the disease free and overall survival of patients.

  2. Estrogens increase expression of bone morphogenetic protein 8b in brown adipose tissue of mice

    NARCIS (Netherlands)

    A. Grefhorst (Aldo); J.C. van den Beukel (Johanna); A.F. van Houten (A.); J. Steenbergen (Jacobie); J.A. Visser (Jenny); A.P.N. Themmen (Axel)

    2015-01-01

    textabstractBackground: In mammals, white adipose tissue (WAT) stores fat and brown adipose tissue (BAT) dissipates fat to produce heat. Several studies showed that females have more active BAT. Members of the bone morphogenetic protein (BMP) and fibroblast growth factor (FGF) families are expressed

  3. Effects of Osseointegration by Bone Morphogenetic Protein-2 on Titanium Implants In Vitro and In Vivo

    OpenAIRE

    Fu-Yuan Teng; Wen-Cheng Chen; Yin-Lai Wang; Chun-Cheng Hung; Chun-Chieh Tseng

    2016-01-01

    This study designed a biomimetic implant for reducing healing time and achieving early osseointegration to create an active surface. Bone morphogenetic protein-2 (BMP-2) is a strong regulator protein in osteogenic pathways. Due to hardly maintaining BMP-2 biological function and specificity, BMP-2 efficient delivery on implant surfaces is the main challenge for the clinic application. In this study, a novel method for synthesizing functionalized silane film for superior modification with BMP-...

  4. Protein PocketViewer: A Web-Service Based Interface for Protein Pocket Extraction and Visualization

    Directory of Open Access Journals (Sweden)

    Xiaoyu Zhang

    2013-09-01

    Full Text Available One important problem in bioinformatics is to study pockets or tunnels within the protein structure. These pocket or tunnel regions are significant because they indicate areas of ligand binding or enzymatic reactions, and tunnels are often solvent ion conductance areas. The Protein Pocket Viewer (PPV is a web interface that allows the user to extract and visualize the protein pockets in a browser, based on the algorithm in [1]. The PPV packaged the pocket extraction executable as a web service, and made it accessible to all users with the Internet access and a modern java enabled browser. The PPV employed the Model2design pattern, which led to a loosely coupled implementation that is more robust and easier to maintain. It consists of a client web interface for user inputs and visualization, a middle-layer for controlling the flow, and the backend web services performing the actual CPU-intensive computation. The PPV web client consists of multiple window regions, with each region providing differing views of the protein, pockets and related information. For a more responsive user experience, the PPV web client employs AJAX for asynchronous execution of long running tasks, like protein pocket extraction.

  5. Improvement of protein extraction from sunflower meal by hydrolysis with alcalase

    OpenAIRE

    Yust, María del Mar; Pedroche, Joaquín; Megías, Cristina; Girón-Calle, Julio; Alaíz Barragán, Manuel; Millán, Francisco; Vioque, Javier

    2003-01-01

    Extraction of proteins from defatted sunflower meal has been improved by addition of the protease alcalase during alkaline extraction. This method offers several additional advantages as compared to the traditional alkaline extraction without alcalase, which is usually carried out after a sedimentation/flotation step to remove the lignocellulosic fraction. As compared to extraction without alcalase, addition of 0.1% (v/v) alcalase improved the yield of protein extraction from 57.5% to 8...

  6. Improvement of protein extraction from sunflower meal by hydrolysis with alcalase

    OpenAIRE

    Vioque, J.; Millán, F.; Alaiz, M.; Girón-Calle, J.; Megías, C.; Pedroche, J.; Yust, M. M.

    2003-01-01

    Extraction of proteins from defatted sunflower meal has been improved by addition of the protease alcalase during alkaline extraction. This method offers several additional advantages as compared to the traditional alkaline extraction without alcalase, which is usually carried out after a sedimentation/flotation step to remove the lignocellulosic fraction. As compared to extraction without alcalase, addition of 0.1% (v/v) alcalase improved the yield of protein extraction from 57.5% to 87.4%, ...

  7. PIPE: a protein-protein interaction passage extraction module for BioCreative challenge.

    Science.gov (United States)

    Chang, Yung-Chun; Chu, Chun-Han; Su, Yu-Chen; Chen, Chien Chin; Hsu, Wen-Lian

    2016-01-01

    Identifying the interactions between proteins mentioned in biomedical literatures is one of the frequently discussed topics of text mining in the life science field. In this article, we propose PIPE, an interaction pattern generation module used in the Collaborative Biocurator Assistant Task at BioCreative V (http://www.biocreative.org/) to capture frequent protein-protein interaction (PPI) patterns within text. We also present an interaction pattern tree (IPT) kernel method that integrates the PPI patterns with convolution tree kernel (CTK) to extract PPIs. Methods were evaluated on LLL, IEPA, HPRD50, AIMed and BioInfer corpora using cross-validation, cross-learning and cross-corpus evaluation. Empirical evaluations demonstrate that our method is effective and outperforms several well-known PPI extraction methods. DATABASE URL. PMID:27524807

  8. Preparation of denatured protein bone sterilized with gamma radiation; Preparacion de hueso desproteinizado esterilizado con radiacion gamma

    Energy Technology Data Exchange (ETDEWEB)

    Luna Z, D. [ININ, 52045 Ocoyoacac, Estado de Mexico (Mexico)]. e-mail: dlz@nuclear.inin.mx

    2005-07-01

    The bone is one of the tissues more transplanted in the entire world by that the bone necessity for transplant every day becomes bigger. In the Bank of tissues Radio sterilized of the ININ the amnion and the pig skin are routinely processed. The tissue with which will be continued is with bone. Due to that in our country it doesn't have enough bone of human origin for the necessities required in the bone transplant, an option is the bone of bovine. Of this bone one can obtain denatured protein bone, with the same characteristics of the denatured protein human bone, the one which has been proven that it has good acceptance and incorporation in the human body when is transplanted. The method for the obtaining of the denatured protein bone of bovine, with the confirmation of the final product by means of X-ray diffraction is described. The radiosterilization of this bone with gamma rays and the determination of the lead content. (Author)

  9. O-glycosylation of protein subpopulations in alcohol-extracted rice proteins.

    Science.gov (United States)

    Kilcoyne, Michelle; Shah, Miti; Gerlach, Jared Q; Bhavanandan, Veer; Nagaraj, Vinay; Smith, Amy D; Fujiyama, Kazuhito; Sommer, Ulf; Costello, Catherine E; Olszewski, Neil; Joshi, Lokesh

    2009-02-15

    Mucin-type O-glycosylation has been well characterized in mammalian systems but not in plants. In this study, the purified alcohol-soluble, non-reduced protein (prolamin) fraction from rice seed was investigated for the occurrence of O-linked oligosaccharides. As storage prolamins are unlikely to be O-glycosylated, any O-glycosylation found was likely to belong to co-extracted proteins, whether because of association with the protein body or solubility. SDS-PAGE and MS analyses revealed 14 and 16kDa protein families in fractions that bound to the lectins peanut agglutinin (PNA), Vicia villosa lectin (VVL) and Jacalin, indicative of the presence of O-linked saccharides. Enzymatic cleavage, fluorescent labeling and high-performance liquid chromatography (HPLC) analysis demonstrated a peak consistent with Gal-beta-(1-->3)-GalNAc, with similar MS/MS fragmentation. Additionally, upon chemical analysis, a GlcNAc-containing O-linked carbohydrate moiety was discovered. Protein blotting with anti-O-GlcNAc antibody (clone CTD110.6) was positive in a subpopulation of the 14kDa alcohol-soluble protein fraction, but a hot capping experiment was negative. Therefore, the GlcNAc residue in this case is unlikely to be terminal. Additionally, a positive reaction with CTD110.6mAb cannot be taken as absolute proof of O-GlcNAc modification and further confirmatory experiments should be employed. We hypothesize that O-glycosylation may contribute to protein functionality or regulation. Further investigation is required to identify the specific proteins with these modifications. This 'reverse' approach could lead to the identification of proteins involved in mRNA targeting, signaling, translation, anchoring or maintenance of translational quiescence and may be applied to germinating rice seed extracts for further elucidation of protein function and regulation. PMID:18639953

  10. Obif, a Transmembrane Protein, Is Required for Bone Mineralization and Spermatogenesis in Mice.

    Directory of Open Access Journals (Sweden)

    Koji Mizuhashi

    Full Text Available Various kinds of transmembrane and secreted proteins play pivotal roles in development through cell-cell communication. We previously reported that Obif (Osteoblast induction factor, Tmem119, encoding a single transmembrane protein, is expressed in differentiating osteoblasts, and that Obif-/- mice exhibit significantly reduced bone volume in the femur. In the current study, we characterized the Obif protein and further investigated the biological phenotypes of a variety of tissues in Obif-/- mice.First, we found that O-glycosylation of the Obif protein occurs at serine residue 36 in the Obif extracellular domain. Next, we observed that Obif-/- mice exhibit bone dysplasia in association with significantly increased osteoid volume per osteoid surface (OV/OS and osteoid maturation time (Omt, and significantly decreased mineral apposition rate (MAR and bone formation rate per bone surface (BFR/BS. In addition, we observed that Obif-/- mice show a significant decrease in testis weight as well as in sperm number. By histological analysis, we found that Obif is expressed in spermatocytes and spermatids in the developing testis and that spermatogenesis is halted at the round spermatid stage in the Obif-/- testis that lacks sperm. However, the number of litters fathered by male mice was slightly reduced in Obif-/- mice compared with wild-type mice, although this was not statistically significant.Our results, taken together with previous observations, indicate that Obif is a type Ia transmembrane protein whose N-terminal region is O-glycosylated. In addition, we found that Obif is required for normal bone mineralization and late testicular differentiation in vivo. These findings suggest that Obif plays essential roles in the development of multiple tissues.

  11. [Ectopic osteogenesis in vivo using bone morphogenetic protein-2 derived peptide loaded biodegradable hydrogel].

    Science.gov (United States)

    Zhao, Jingjing; Fang, Zhenhua; Huang, Ruokun; Xiao, Kai; Li, Jing; Xie, Ming; Kan, Wusheng

    2014-08-01

    We investigated the development of an injectable, biodegradable hydrogel composite of poly(trimethylene carbonate)-F127-poly(trimethylene carbonate)(PTMC11-F127-PTMC11 )loaded with bone morphogenetic protein-2 (BMP-2) derived peptide P24 for ectopic bone formation in vivo and evaluated its release kinetics in vitro. Then we evaluated P24 peptide release kinetics from different concentration of PTMC11-F127-PTMC11 hydrogel in vitro using bicinchoninic acid (BCA)assay. P24/ PTMC11-F127-PTMC11 hydrogel was implanted into each rat's erector muscle of spine and ectopic bone formation of the implanted gel in vivo was detected by hematoxylin and eosin stain (HE). PTMC11-F127-PTMC11 hydrogel with concentration more than 20 percent showed sustained slow release for one month after the initial burst release. Bone trabeculae surround the P24/ PTMC11-F127-PTMC11 hydrogel was shown at the end of six weeks by hematoxylin and eosin stain. These results indicated that encapsulated bone morphogenetic protein (BMP-2) derived peptide P24 remained viable in vivo, thus suggesting the potential of PTMC11-F127-PT- MC11 composite hydrogels as part of a novel strategy for localized delivery of bioactive molecules. PMID:25508424

  12. Bone morphogenetic protein 2 and decorin expression in old fracture fragments and surrounding tissues.

    Science.gov (United States)

    Han, X G; Wang, D K; Gao, F; Liu, R H; Bi, Z G

    2015-01-01

    Bone morphogenetic protein 2 (BMP-2) can promote fracture healing. Although the complex role BMP-2 in bone formation is increasingly understood, the role of endogenous BMP-2 in nonunion remains unclear. Decorin (DCN) can promote the formation of bone matrix and calcium deposition to control bone morphogenesis. In this study, tissue composition and expression of BMP-2 and DCN were detected in different parts of old fracture zones to explore inherent anti-fibrotic ability and osteogenesis. Twenty-three patients were selected, including eight cases of delayed union and 15 cases of nonunion. Average duration of delayed union or nonunion was 15 months. Fracture fragments and surrounding tissues, including bone grafts, marrow cavity contents, and sticking scars, were categorically sampled during surgery. Through observation and histological testing, component comparisons were made between fracture fragments and surrounding tissue. The expression levels of DCN and BMP-2 in different tissues were detected by immunohistochemical staining and real-time polymerase chain reaction. The expression of DCN and BMP- 2 in different parts of the nonunion area showed that, compared with bone graft and marrow cavity contents, sticking scars had the highest expression of BMP-2. Compared with the marrow cavity contents and sticking scars, bone grafts had the highest expression of DCN. The low antifibrotic and osteogenic activity of the nonunion area was associated with non-co-expression of BMP-2 and DCN. Therefore, the co-injection of osteogenic factor BMP and DCN into the nonunion area can improve the induction of bone formation and enhance the conversion of the old scar, thereby achieving better nonunion treatment. PMID:26400336

  13. Hydrolyzing Condition and Immunocompetence of Sheep Bone Protein Enzymatic Lysates

    Institute of Scientific and Technical Information of China (English)

    YANG Hua; LIU Yu-hua; MA Li-zhen; KONG Bao-hua

    2009-01-01

    Utilizing collagen of sheep bone as material to get immunocompetent peptide, enzymatic hydrolysis conditions were optimized using quadratic regression general rotation design. The effect of temperature (T), time (t), enzyme/substrate (E/S) ratio, and substrate concentration (S) on the amount of tricarboxylix acid cycle (TCA) soluble peptides were investigated. The content of soluble peptide in the acquisition was measured by Folin-hydrozybebzebe method, and the correlation between soluble peptide content and immunocompetence was analyzed by SAS software. The best enzymatic hydrolysis condition was gotten from Design Expert 7.1.2 software. The optimal condition under which immunocompetent peptides could be prepared was 1 576 U g-1 (E/S), 64.05℃ (T), 0.271 kg L-1(S), and 7.22 h (t). The correlation coefficient between TCA soluble peptides and the immuneocompetence was 0.045 < r0.05= 0.355, which indicated that they had no significant correlation. The result showed that the soluble peptide contained immunocompetent peptides which content was independent of immunocompetence in the hydrolasates.

  14. Demineralized dentin matrix combined with recombinant human bone morphogenetic protein-2 in rabbit calvarial defects

    OpenAIRE

    Um, In-Woong; Hwang, Suk-Hyun; Kim, Young-Kyun; Kim, Moon-Young; Jun, Sang-Ho; Ryu, Jae-Jun; Jang, Hyon-Seok

    2016-01-01

    Objectives The aim of this study was to compare the osteogenic effects of demineralized dentin matrix (DDM) combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) in rabbit calvarial defects with DDM and anorganic bovine bone (ABB) combined with rhBMP-2. Materials and Methods Four round defects with 8-mm diameters were created in each rabbit calvaria. Each defect was treated with one of the following: 1) DDM, 2) ABB/rhBMP-2, or 3) DDM/rhBMP-2. The rhBMP-2 was combined with DDM...

  15. Influence of ethylene oxide sterilization on the activity of native reindeer bone morphogenetic protein

    OpenAIRE

    Pekkarinen, T. (Tarmo); Hietala, O.; Lindholm, T. S.; Jalovaara, P.

    2003-01-01

    We studied the effects of ethylene oxide sterilization (Steri-Vac 4XL, temperature 29°C, exposure time 4 h 10 min, ethylene oxide concentration 860 mg/l) on the osteoinductivity of partially purified native reindeer bone morphogenetic protein (BMP) in a hind leg muscle pouch model of male NMRI mice. BMP was administered in implants containing 3 mg in a collagen carrier. Implants without sterilization and without BMP served as controls. New bone formation was evaluated based on the calcium yie...

  16. Biomineralization of Engineered Spider Silk Protein-Based Composite Materials for Bone Tissue Engineering

    Directory of Open Access Journals (Sweden)

    John G. Hardy

    2016-07-01

    Full Text Available Materials based on biodegradable polyesters, such as poly(butylene terephthalate (PBT or poly(butylene terephthalate-co-poly(alkylene glycol terephthalate (PBTAT, have potential application as pro-regenerative scaffolds for bone tissue engineering. Herein, the preparation of films composed of PBT or PBTAT and an engineered spider silk protein, (eADF4(C16, that displays multiple carboxylic acid moieties capable of binding calcium ions and facilitating their biomineralization with calcium carbonate or calcium phosphate is reported. Human mesenchymal stem cells cultured on films mineralized with calcium phosphate show enhanced levels of alkaline phosphatase activity suggesting that such composites have potential use for bone tissue engineering.

  17. Pomegranate Peel Extract Prevents Bone Loss in a Preclinical Model of Osteoporosis and Stimulates Osteoblastic Differentiation in Vitro

    Directory of Open Access Journals (Sweden)

    Mélanie Spilmont

    2015-11-01

    Full Text Available The nutritional benefits of pomegranate have attracted great scientific interest. The pomegranate, including the pomegranate peel, has been used worldwide for many years as a fruit with medicinal activity, mostly antioxidant properties. Among chronic diseases, osteoporosis, which is associated with bone remodelling impairment leading to progressive bone loss, could eventually benefit from antioxidant compounds because of the involvement of oxidative stress in the pathogenesis of osteopenia. In this study, with in vivo and ex vivo experiments, we investigated whether the consumption of pomegranate peel extract (PGPE could limit the process of osteopenia. We demonstrated that in ovariectomized (OVX C57BL/6J mice, PGPE consumption was able to significantly prevent the decrease in bone mineral density (−31.9%; p < 0.001 vs. OVX mice and bone microarchitecture impairment. Moreover, the exposure of RAW264.7 cells to serum harvested from mice that had been given a PGPE-enriched diet elicited reduced osteoclast differentiation and bone resorption, as shown by the inhibition of the major osteoclast markers. In addition, PGPE appeared to substantially stimulate osteoblastic MC3T3-E1 alkaline phosphatase (ALP activity at day 7, mineralization at day 21 and the transcription level of osteogenic markers. PGPE may be effective in preventing the bone loss associated with ovariectomy in mice, and offers a promising alternative for the nutritional management of this disease.

  18. Bone Marrow Mesenchymal Stem Cells Expressing Baculovirus-Engineered Bone Morphogenetic Protein-7 Enhance Rabbit Posterolateral Fusion.

    Science.gov (United States)

    Liao, Jen-Chung

    2016-01-01

    Previous studies have suggested that bone marrow-derived mesenchymal stem cells (BMDMSCs) genetically modified with baculoviral bone morphogenetic protein-2 (Bac-BMP-2) vectors could achieve successful fusion in a femur defect model or in a spinal fusion model. In this study, BMDMSCs expressing BMP-7 (Bac-BMP-7-BMDMSCs) were generated. We hypothesized that Bac-BMP-7-BMDMSCs could secrete more BMP-7 than untransduced BMDMSCs in vitro and achieve spinal posterolateral fusion in a rabbit model. Eighteen rabbits underwent posterolateral fusion at L4-5. Group I (n = 6) was implanted with collagen-β-tricalcium phosphate (TCP)-hydroxyapatite (HA), Group II (n = 6) was implanted with collagen-β-TCP-HA plus BMDMSCs, and Group III (n = 6) was implanted with collagen-β-TCP-HA plus Bac-BMP-7-BMDMSCs. In vitro production of BMP-7 was quantified with an enzyme-linked immunosorbent assay (ELISA). Spinal fusion was examined using computed tomography (CT), manual palpation, and histological analysis. ELISA demonstrated that Bac-BMP-7-BMDMSCs produced four-fold to five-fold more BMP-7 than did BMDMSCs. In the CT results, 6 fused segments were observed in Group I (50%, 6/12), 8 in Group II (67%, 8/12), and 12 in Group III (100%, 12/12). The fusion rate, determined by manual palpation, was 0% (0/6) in Group I, 0% (0/6) in Group II, and 83% (5/6) in Group III. Histology showed that Group III had more new bone and matured marrow formation. In conclusion, BMDMSCs genetically transduced with the Bac-BMP-7 vector could express more BMP-7 than untransduced BMDMSCs. These Bac-BMP-7-BMDMSCs on collagen-β-TCP-HA scaffolds were able to induce successful spinal fusion in rabbits. PMID:27399674

  19. Bone Marrow Mesenchymal Stem Cells Expressing Baculovirus-Engineered Bone Morphogenetic Protein-7 Enhance Rabbit Posterolateral Fusion

    Directory of Open Access Journals (Sweden)

    Jen-Chung Liao

    2016-07-01

    Full Text Available Previous studies have suggested that bone marrow-derived mesenchymal stem cells (BMDMSCs genetically modified with baculoviral bone morphogenetic protein-2 (Bac-BMP-2 vectors could achieve successful fusion in a femur defect model or in a spinal fusion model. In this study, BMDMSCs expressing BMP-7 (Bac-BMP-7-BMDMSCs were generated. We hypothesized that Bac-BMP-7-BMDMSCs could secrete more BMP-7 than untransduced BMDMSCs in vitro and achieve spinal posterolateral fusion in a rabbit model. Eighteen rabbits underwent posterolateral fusion at L4-5. Group I (n = 6 was implanted with collagen-β-tricalcium phosphate (TCP-hydroxyapatite (HA, Group II (n = 6 was implanted with collagen-β-TCP-HA plus BMDMSCs, and Group III (n = 6 was implanted with collagen-β-TCP-HA plus Bac-BMP-7-BMDMSCs. In vitro production of BMP-7 was quantified with an enzyme-linked immunosorbent assay (ELISA. Spinal fusion was examined using computed tomography (CT, manual palpation, and histological analysis. ELISA demonstrated that Bac-BMP-7-BMDMSCs produced four-fold to five-fold more BMP-7 than did BMDMSCs. In the CT results, 6 fused segments were observed in Group I (50%, 6/12, 8 in Group II (67%, 8/12, and 12 in Group III (100%, 12/12. The fusion rate, determined by manual palpation, was 0% (0/6 in Group I, 0% (0/6 in Group II, and 83% (5/6 in Group III. Histology showed that Group III had more new bone and matured marrow formation. In conclusion, BMDMSCs genetically transduced with the Bac-BMP-7 vector could express more BMP-7 than untransduced BMDMSCs. These Bac-BMP-7-BMDMSCs on collagen-β-TCP-HA scaffolds were able to induce successful spinal fusion in rabbits.

  20. Enzymatic aqueous technology for simultaneous coconut protein and oil extraction

    Directory of Open Access Journals (Sweden)

    Coelho, Maria A. Z.

    2003-03-01

    Full Text Available In this study, the following commercial enzymes were evaluated in the enzymatic extraction of oil and protein from coconut: Celluclast, Termamyl, Viscozyme, Neutrase y Protease. Preliminary experiments were carried out for selection of enzymes, enzyme concentration and incubation time. In order to verify the effects of pH and water/substrate ratio, a response surface methodology was applied based in a 32 full factorial experimental design. The pH is the most meaningful parameter on oil and protein extraction yields, with a significance level higher than 90%. The maximal extraction yields of oil and protein emulsion (83% was reached using Viscozyme L and subsequently Neutrase 1.5 MG at concentrations of 0.6% (w/w and 0.3% (w/w, respectively, total incubation time of 60 minutes, substrate/water ratio 1:6 and pH around 7 (no adjustment.En este trabajo diferentes enzimas hidroliticos fueron usados para la extracción de aceite y proteína de coco: Celluclast, Termamyl, Viscozyme, Neutrase y Protease producidos por Novo Nordisk. Experimientos preliminares fueron encaminados a la selección de los extractos enzimaticos, de las concentraciones de las enzimas y de los tiempos de incubación. La técnica de análisis de varianza, mediante un delineamiento factorial en tres niveles, fue usada para la optimización de los procesos con relación a los parámetros pH y razón sustrato/agua. Comparando con la extracción acuosa todos los enzimas hidroliticos incrementan la extracción de aceite y proteína vegetal de coco. Los coeficientes de correlación muestran que el pH fué el parámetro mas significativo (pViscozyme y Neutrase 0.6 % (w/w y 0.3 % (w/w, respectivamente, tiempo de incubación 30 minutos, razón sustrato/agua 1:6 y pH 7.

  1. Potential bone-inducing activity in vitro of recombinant human bone morphogenetic protein-7 from a CHO expression system

    Institute of Scientific and Technical Information of China (English)

    LI Xiao-yan; SHI Wei-wei; WANG Hao; LI Bo-hua; YANG Yang; TAN Min; XUE Jing-ya; GUO Ya-jun

    2005-01-01

    Objective: To express the recombinant human bone morphogenetic protein-7 (rhBMP-7) in Chinese hamster ovary(CHO) cells, and to establish the in vitro biological activity assay of rhBMP-7.Methods: Human BMP-7 cDNA was subcloned into p114 mammalian expression vector and transfected to CHO cells by using the Lipofectamine2000 transfection method. CHO cell supernatants were harvested and analyzed to identify the molecule mass of secreted rhBMP-7 and examine its biological activity in vitro to stimulate the synthesis of alkaline phophatase(ALP), a characteristic of osteoblast phenotypes. Results:rhBMP-7 was produced stably in CHO cells, as a processed mature disulfide-linked homodimer, with an apparent molecular mass of 36 000. Examination of the rhBMP-7 biological activity showed that rhBMP-7 specifically stimulated the production of ALP(4-fold increase at 100 ng of rhBMP-7/ml). Conclusion: The rhBMP-7 from CHO expression system has significant biological activity in induction of osteoblast phenotype, which demonstrates rhBMP-7 has the potential bone regeneration activity.

  2. Effects of ionizing radiation on wound healing of alveolar bone socket after extraction of rat maxillary molars

    International Nuclear Information System (INIS)

    The purpose of this study was to examine the effects of radiation on the healing process of tooth extraction wounds. X-ray doses of 10 Gy or 20 Gy were delivered, once, to the maxillofacial area of Wistar-strain rats. Then, 24 hours after irradiation, the maxillary first molars were extracted bilaterally. The animals were sacrificed 3, 7, 10, 14, 21, 42, and 84 days after tooth extraction, and the maxilla were sliced, to make thin sections. These specimens were then double stained with alkaline phosphatase (ALP) and tartrate resistant acid phosphatase (TRAP). The ratio of bone area to socket area (bone formation ratio), the ratio of bone length to ALP positive area length (ALP positive ratio), and the number of TRAP-positive cells, were evaluated. The results showed: The bone formation ratios at days 3 and 7 after tooth extraction were significantly low in both irradiation groups, compared with those for the non-irradiation group. The ALP positive reaction ratio peaked 7 days after in the non-irradiation group. In both irradiation groups, the ratios that were worked out at 3 days and 7 days after were significantly lower than those in the non-irradiation group. There was no significant difference in the number of TRAP-positive cells between the non-irradiation group and the 10 Gy irradiation group. In the 20 Gy irradiation group, the TRAP-positive cell count plummeted to a significantly low level at 3 days after tooth extraction, compared with that in the non-irradiation group. (author)

  3. Extraction, composition and functional properties of pennycress (Thlaspi arvense L.) press cake protein

    Science.gov (United States)

    This study compared two methods for extracting the protein in pennycress (Thlaspi arvense L.) press cake and determined the composition and functional properties of the protein products. Proteins in pennycress press cake were extracted by using the conventional alkali solubilization-acid precipitati...

  4. Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics.

    Science.gov (United States)

    Wu, Huifeng; Ji, Chenglong; Wei, Lei; Zhao, Jianmin

    2013-11-01

    In ecotoxicoproteomics, an accurate and reproducible extraction of proteins is a critical step for 2DE analysis and further protein identification using MS. The criteria for the assessment of protein extraction quality include protein yield, protein spots resolved in a 2DE gel, matched protein spots in replicate gels, reproducibility, and compatibility with MS. In this work, we evaluated three protein extraction systems, straightforward lysis buffer, trichloroacetic acid-acetone, and TRIzol reagent with some modifications, for the protein extraction from three animal species including mussel Mytilus galloprovincialis, flounder Paralichthys olivaceus, and polychaete Nereis diversicolor used in marine ecotoxicology. Our results indicated that these methods could extract significantly different protein profiles. The method using TRIzol reagent resulted in the most matched protein spots resolved in four replicate 2DE gels and highest reproducibilities for the gill of M. galloprovincialis and liver of P. olivaceus. However, a modified trichloroacetic acid-acetone solvent system was best for the whole soft tissue of N. diversicolor. This work provides the fundamental information of the extraction quality of protein extraction protocols from different marine animals, which may facilitate the selection of a suitable protein extraction protocol for ecotoxicoproteomics. PMID:24106175

  5. Comparison of Two Methods for the Extraction of Fractionated Rice Bran Protein

    OpenAIRE

    Changyuan Wang; Dan Li; Feng Xu; Tianshu Hao; Min Zhang

    2014-01-01

    Two different methods for extracting fractionated rice bran protein (FRBP) from defatted rice bran were investigated according to the solubility of protein in different extraction solvents. The yields of the obtained proteins and their purity were first compared. Sodium dodecyl sulfate polyacrylamide gel electrophoresis, differential scanning calorimetry, protein surface hydrophobicity, and protein secondary molecular structure analyses were subsequently applied to identify and compare the co...

  6. A comprehensive benchmark of kernel methods to extract protein-protein interactions from literature.

    Directory of Open Access Journals (Sweden)

    Domonkos Tikk

    Full Text Available The most important way of conveying new findings in biomedical research is scientific publication. Extraction of protein-protein interactions (PPIs reported in scientific publications is one of the core topics of text mining in the life sciences. Recently, a new class of such methods has been proposed - convolution kernels that identify PPIs using deep parses of sentences. However, comparing published results of different PPI extraction methods is impossible due to the use of different evaluation corpora, different evaluation metrics, different tuning procedures, etc. In this paper, we study whether the reported performance metrics are robust across different corpora and learning settings and whether the use of deep parsing actually leads to an increase in extraction quality. Our ultimate goal is to identify the one method that performs best in real-life scenarios, where information extraction is performed on unseen text and not on specifically prepared evaluation data. We performed a comprehensive benchmarking of nine different methods for PPI extraction that use convolution kernels on rich linguistic information. Methods were evaluated on five different public corpora using cross-validation, cross-learning, and cross-corpus evaluation. Our study confirms that kernels using dependency trees generally outperform kernels based on syntax trees. However, our study also shows that only the best kernel methods can compete with a simple rule-based approach when the evaluation prevents information leakage between training and test corpora. Our results further reveal that the F-score of many approaches drops significantly if no corpus-specific parameter optimization is applied and that methods reaching a good AUC score often perform much worse in terms of F-score. We conclude that for most kernels no sensible estimation of PPI extraction performance on new text is possible, given the current heterogeneity in evaluation data. Nevertheless, our study

  7. Nanofibrous yet injectable polycaprolactone-collagen bone tissue scaffold with osteoprogenitor cells and controlled release of bone morphogenetic protein-2

    Energy Technology Data Exchange (ETDEWEB)

    Subramanian, Gayathri; Bialorucki, Callan [Department of Bioengineering, College of Engineering, University of Toledo, Toledo, OH 43606 (United States); Yildirim-Ayan, Eda, E-mail: eda.yildirimayan@utoledo.edu [Department of Bioengineering, College of Engineering, University of Toledo, Toledo, OH 43606 (United States); Department of Orthopaedic Surgery, University of Toledo Medical Center, Toledo, OH 43614 (United States)

    2015-06-01

    In this work, we developed a nanofibrous, yet injectable orthobiologic tissue scaffold that is capable of hosting osteoprogenitor cells and controlling kinetic release profile of the encapsulated pro-osteogenic factor without diminishing its bioactivity over 21 days. This innovative injectable scaffold was synthesized by incorporating electrospun and subsequently O{sub 2} plasma-functionalized polycaprolactone (PCL) nanofibers within the collagen type-I solution along with MC3T3-E1 cells (pre-osteoblasts) and bone morphogenetic protein-2 (BMP2). Through changing the PCL nanofiber concentration within the injectable scaffolds, we were able to tailor the mechanical strength, protein retention capacity, bioactivity preservation, and osteoinductive potential of the scaffolds. The nanofibrous internal structure of the scaffold allowed us to use a low dose of BMP2 (200 ng/ml) to achieve osteoblastic differentiation in in vitro culture. The osteogenesis capacity of the injectable scaffolds were evaluated though measuring MC3T3-E1 cell proliferation, ALP activity, matrix mineralization, and early- and late-osteoblast specific gene expression profiles over 21 days. The results demonstrated that the nanofibrous injectable scaffold provides not only an osteoinductive environment for osteoprogenitor cells to differentiate, but also a suitable biomechanical and biochemical environment to act as a reservoir for osteogenic factors with controlled release profile. - Highlights: • Injectable nanofibrous scaffold with osteoprogenitor cells and BMP2 was synthesized. • PCL nanofiber concentration within collagen scaffold affected the BMP2 retention and bioactivity. • Optimal PCL concentration was identified for mechanical stability, injectability, and osteogenic activity. • Scaffolds exhibited long-term osteoinductive capacity for bone repair and regeneration.

  8. Nanofibrous yet injectable polycaprolactone-collagen bone tissue scaffold with osteoprogenitor cells and controlled release of bone morphogenetic protein-2

    International Nuclear Information System (INIS)

    In this work, we developed a nanofibrous, yet injectable orthobiologic tissue scaffold that is capable of hosting osteoprogenitor cells and controlling kinetic release profile of the encapsulated pro-osteogenic factor without diminishing its bioactivity over 21 days. This innovative injectable scaffold was synthesized by incorporating electrospun and subsequently O2 plasma-functionalized polycaprolactone (PCL) nanofibers within the collagen type-I solution along with MC3T3-E1 cells (pre-osteoblasts) and bone morphogenetic protein-2 (BMP2). Through changing the PCL nanofiber concentration within the injectable scaffolds, we were able to tailor the mechanical strength, protein retention capacity, bioactivity preservation, and osteoinductive potential of the scaffolds. The nanofibrous internal structure of the scaffold allowed us to use a low dose of BMP2 (200 ng/ml) to achieve osteoblastic differentiation in in vitro culture. The osteogenesis capacity of the injectable scaffolds were evaluated though measuring MC3T3-E1 cell proliferation, ALP activity, matrix mineralization, and early- and late-osteoblast specific gene expression profiles over 21 days. The results demonstrated that the nanofibrous injectable scaffold provides not only an osteoinductive environment for osteoprogenitor cells to differentiate, but also a suitable biomechanical and biochemical environment to act as a reservoir for osteogenic factors with controlled release profile. - Highlights: • Injectable nanofibrous scaffold with osteoprogenitor cells and BMP2 was synthesized. • PCL nanofiber concentration within collagen scaffold affected the BMP2 retention and bioactivity. • Optimal PCL concentration was identified for mechanical stability, injectability, and osteogenic activity. • Scaffolds exhibited long-term osteoinductive capacity for bone repair and regeneration

  9. Chemically-Conjugated Bone Morphogenetic Protein-2 on Three-Dimensional Polycaprolactone Scaffolds Stimulates Osteogenic Activity in Bone Marrow Stromal Cells

    OpenAIRE

    Zhang, Huina; Migneco, Francesco; Lin, Chia-Ying; Hollister, Scott J.

    2010-01-01

    Poly(ε-caprolactone) (PCL) has received considerable attention in bone tissue engineering. However, the lack of osteoinductive ability of PCL limits its application. The aim of this study was to directly attach bone morphogenetic protein-2 (BMP-2) to PCL scaffolds by a crosslinking conjugation method and to investigate whether the bound BMP-2 maintained bioactivity in vitro. Immunofluorescent staining against BMP-2 and quantitative enzyme-linked immunosorbent assay measurements demonstrated t...

  10. Improved Bone Formation in Osteoporotic Rabbits with the Bone Morphogenetic Protein-2 (rhBMP-2) Coated Titanium Screws Which Were Coated By Using Plasma Polymerization Technique

    OpenAIRE

    Salih Gulsen; Dilek Cokeliler; Hilal Goktas; Aysu Kucukturhan; Bilgehan Ozcil; Hakan Caner

    2014-01-01

    Delaying of bone fusion in osteoporotic patients underwent spinal stabilization surgery leads to screw loosening, and this causes pseudoarticulation, mobility and fibrosis at vertebral segments. To prevent these complications, the screws coated with recombinant bone morphogenetic protein-2 (rhBMP-2) could be used. To verify this hypothesis, we coated 5 Titanium screws with rhBMP-2 using plasma polymerization method, and also used 10 uncoated screws for making comparison between coated and unc...

  11. Challenges of protein extraction from recalcitrant plant tissues for proteomics

    Science.gov (United States)

    Proteins play an important role in several biological processes. Proteomics encompasses basically four principal applications, namely protein mining, protein expression profiling, protein-network mapping and mapping of protein modifications. The results in these applications depend mostly on the c...

  12. Bone Morphogenetic Proteins in Craniofacial Surgery: Current Techniques, Clinical Experiences, and the Future of Personalized Stem Cell Therapy

    Directory of Open Access Journals (Sweden)

    Kristofer E. Chenard

    2012-01-01

    Full Text Available Critical-size osseous defects cannot heal without surgical intervention and can pose a significant challenge to craniofacial reconstruction. Autologous bone grafting is the gold standard for repair but is limited by a donor site morbidity and a potentially inadequate supply of autologous bone. Alternatives to autologous bone grafting include the use of alloplastic and allogenic materials, mesenchymal stem cells, and bone morphogenetic proteins. Bone morphogenetic proteins (BMPs are essential mediators of bone formation involved in the regulation of differentiation of osteoprogenitor cells into osteoblasts. Here we focus on the use of BMPs in experimental models of craniofacial surgery and clinical applications of BMPs in the reconstruction of the cranial vault, palate, and mandible and suggest a model for the use of BMPs in personalized stem cell therapies.

  13. Protein extraction from heat-stabilized defatted rice bran. 1. Physical processing and enzyme treatments.

    Science.gov (United States)

    Tang, Shanhu; Hettiarachchy, Navam S; Shellhammer, Thomas H

    2002-12-01

    Physical processing with or without enzyme treatments on protein extraction from heat-stabilized defatted rice bran (HDRB) was evaluated. Freeze-thaw, sonication, high-speed blending, and high-pressure methods extracted 12%, 15%, 16%, and 11% protein, respectively. Sonication (0-100%, 750 W), followed by amylase and combined amylase and protease treatments, extracted 25.6-33.9% and 54.0-57.8% protein, respectively. Blending followed by amylase and protease treatment extracted 5.0% more protein than the nonblended enzymatic treatments. High-pressure treatments, 0-800 MPa, with water or amylase-protease combinations, extracted 10.5-11.1% or 61.8-66.6% protein, respectively. These results suggest that physical processing in combination with enzyme treatments can be effective in extracting protein from HDRB. PMID:12452673

  14. Estrogen modulates the mRNA levels for cancellous bone protein of ovariectomized rats.

    Science.gov (United States)

    Salih, M A; Liu, C C; Arjmandi, B H; Kalu, D N

    1993-12-01

    This study was undertaken to examine the effects of ovariectomy and 17 beta-estradiol (E2) on the gene expression of type 1 collagen, osteocalcin and the protooncogen, c-myc, in cancellous bone. Female Sprague-Dawley rats, aged 95 days, were divided into 4 groups. Group 1 was sham operated and Groups 2-4 were ovariectomized. Groups 3 and 4 received daily injections of 160 ng and 1600 ng E2/kg body weight, respectively. Groups 1 and 2 received the solvent vehicle. All animals were sacrificed after 14 days. The femurs were dissected out and cancellous bone scraped from the distal metaphysis. RNA was isolated from the cancellous bone, immobilized on filters or size-fractionated by agarose gel electrophoresis and adsorbed on filters which were then hybridized with specific cDNA probes. Ovariectomy resulted in a significant increase in the mRNAs of type 1 collagen, osteocalcin and c-myc. The increase was suppressed in animals that received 17 beta-estradiol injections. In addition, ovariectomy caused the expected decrease in cancellous bone in the proximal tibia and increased osteoclast and osteoblast numbers. The ovariectomy-induced changes were prevented by 17 beta-estradiol administration. These findings suggest that the lack of ovarian hormones shortly after ovariectomy up-regulates and estrogen administration down-regulates the expression of important cancellous bone matrix proteins as well as the protooncogen, c-myc. PMID:8148671

  15. Healing patterns of critical size bony defects in rats after grafting with bone substitutes soaked in recombinant human bone morphogenetic protein-2: histological and histometric evaluation.

    Science.gov (United States)

    Mokbel, N; Naaman, N; Nohra, J; Badawi, N

    2013-09-01

    The aim of the study was to evaluate the effect of different bone substitutes soaked in recombinant human bone morphogenetic protein-2 (rhBMP-2) on the healing of critical size defects in calvarial bone. Defects were created in 24 Sprague Dawley rats. The rhBMP-2 was diluted to obtain a final concentration of 0.2mg/ml. Rats were divided into four groups and treated as follows: in the first group the defect was filled with anorganic bovine bone mineral (ABBM) and rhBMP-2, the second group was treated with freeze-dried bone allograft (FDBA) and rhBMP-2, and the third group was treated with autogenous bone (AUTO). In the control group the defects were left untreated. Animals were killed after 8weeks and calcified histological sections prepared. Histometric measurements showed that mean (SD) bone formation was 4.00 (1.69)mm(2) in the ABBM group, 2.56 (1.06)mm(2) in the FDBA group, and 2.30 (0.34)mm(2) in the AUTO group. The difference between the ABBM group and the other 3 groups was significant (p<0.0001) with a mean bone formation of 0.82 (0.25)mm(2) in the control group. There was no significant difference between the FDBA and the AUTO groups (p=0.96). Within the limits of this study we concluded that the addition of rhBMP-2 to bone substitutes was efficacious in regenerating bone in critical size bone defects in calveria in rats. PMID:22939894

  16. Point-counter-point debate: the association between recombinant human bone morphogenetic protein utilization and complications in spine surgery

    OpenAIRE

    Siemionow, Kris; Sundberg, Eric; Tyrakowski, Marcin; Nandyala, Sreeharsha V.; Singh, Kern

    2014-01-01

    Bone morphogenetic proteins (BMPs) have been utilized in spine surgery for over 10 years as a bone graft substitute. Potential BMP-related adverse effects including retrograde ejaculation and heterotopic neuroforaminal bone formation have been described. Additionally, some studies have suggested an association between BMP and cancer. Inconsistencies exist in the published spine literature with regards to the incidence and association of complications with BMP utilization. In a point-counterpo...

  17. Protection of Trabecular Bone in Ovariectomized Rats by Turmeric (Curcuma longa L.) is Dependent on Extract Composition

    OpenAIRE

    Wright, Laura E; Frye, Jennifer B.; Timmermann, Barbara N.; Funk, Janet L.

    2010-01-01

    Extracts prepared from turmeric (Curcuma longa L., [Zingiberaceae]) containing bioactive phenolic curcuminoids were evaluated for bone-protective effects in a hypogonadal rat model of postmenopausal osteoporosis. Three-month female Sprague Dawley rats were ovariectomized (OVX) and treated with a chemically complex turmeric fraction (41% curcuminoids by weight) or a curcuminoid-enriched turmeric fraction (94% curcuminoids by weight), both dosed at 60mg/kg 3x per week, or vehicle alone. Effects...

  18. Epidemiologic trends in the utilization, demographics, and cost of bone morphogenetic protein in spinal fusions

    OpenAIRE

    Louie, Philip K.; Hassanzadeh, Hamid; Singh, Kern

    2014-01-01

    Bone morphogenetic protein (BMP) utilization as an adjunct for spinal arthrodesis has gained considerable momentum among spine surgeons. Despite carrying Food and Drug Administration approval for only single level anterior lumbar interbody fusion from L4-S1, the majority of BMP administration is in “off label” settings. Over the last decade, BMP utilization has increased in all facets of spine surgery with the only exception being the anterior cervical spine, in which a downward trend resulte...

  19. Estrogens increase expression of bone morphogenetic protein 8b in brown adipose tissue of mice

    OpenAIRE

    Grefhorst, Aldo; van den Beukel, Johanna C; van Houten, E Leonie AF; Steenbergen, Jacobie; Visser, Jenny A.; Themmen, Axel PN

    2015-01-01

    Background In mammals, white adipose tissue (WAT) stores fat and brown adipose tissue (BAT) dissipates fat to produce heat. Several studies showed that females have more active BAT. Members of the bone morphogenetic protein (BMP) and fibroblast growth factor (FGF) families are expressed in BAT and are involved in BAT activity. We hypothesized that differential expression of BMPs and FGFs might contribute to sex differences in BAT activity. Methods We investigated the expression of BMPs and FG...

  20. Functional assay, expression of growth factors and proteins modulating bone-arrangement in human osteoblasts seeded on an anorganic bovine bone biomaterial

    Directory of Open Access Journals (Sweden)

    O Trubiani

    2010-07-01

    Full Text Available The basic aspects of bone tissue engineering include chemical composition and geometry of the scaffold design, because it is very important to improve not only cell attachment and growth but especially osteodifferentiation, bone tissue formation, and vascularization. Geistlich Bio-Oss® (GBO is a xenograft consisting of deproteinized, sterilized bovine bone, chemically and physically identical to the mineral phase of human bone.In this study, we investigated the growth behaviour and the ability to form focal adhesions on the substrate, using vinculin, a cytoskeletal protein, as a marker. Moreover, the expression of bone specific proteins and growth factors such as type I collagen, osteopontin, bone sialoprotein, bone morphogenetic protein-2 (BMP-2, BMP-7 and de novo synthesis of osteocalcin in normal human osteoblasts (NHOst seeded on xenogenic GBO were evaluated. Our observations suggest that after four weeks of culture in differentiation medium, the NHOst showed a high affinity for the three dimensional biomaterial; in fact, cellular proliferation, migration and colonization were clearly evident. The osteogenic differentiation process, as demonstrated by morphological, histochemical, energy dispersive X-ray microanalysis and biochemical analysis was mostly obvious in the NHOst grown on three-dimensional inorganic bovine bone biomaterial. Functional studies displayed a clear and significant response to calcitonin when the cells were differentiated. In addition, the presence of the biomaterial improved the response, suggesting that it could drive the differentiation of these cells towards a more differentiated osteogenic phenotype. These results encourage us to consider GBO an adequate biocompatible three-dimensional biomaterial, indicating its potential use for the development of tissue-engineering techniques.

  1. Recombinant Human Bone Morphogenetic Protein-2 in Development and Progression of Oral Squamous Cell Carcinoma.

    Science.gov (United States)

    Zaid, Khaled Waleed; Chantiri, Mansour; Bassit, Ghassan

    2016-01-01

    Bone morphogenetic proteins (BMPs), belonging to the transforming growth factor-β superfamily, regulate many cellular activities including cell migration, differentiation, adhesion, proliferation and apoptosis. Use of recombinant human bone morphogenic protein?2 (rhBMP?2) in oral and maxillofacial surgery has seen a tremendous increase. Due to its role in many cellular pathways, the influence of this protein on carcinogenesis in different organs has been intensively studied over the past decade. BMPs also have been detected to have a role in the development and progression of many tumors, particularly disease-specific bone metastasis. In oral squamous cell carcinoma - the tumor type accounting for more than 90% of head and neck malignancies- aberrations of both BMP expression and associated signaling pathways have a certain relation with the development and progression of the disease by regulating a range of biological functions in the altered cells. In the current review, we discuss the influence of BMPs -especially rhBMP-2- in the development and progression of oral squamous cell carcinoma. PMID:27039814

  2. Identification of Proteins and Peptide Biomarkers for Detecting Banned Processed Animal Proteins (PAPs) in Meat and Bone Meal by Mass Spectrometry.

    Science.gov (United States)

    Marbaix, Hélène; Budinger, Dimitri; Dieu, Marc; Fumière, Olivier; Gillard, Nathalie; Delahaut, Philippe; Mauro, Sergio; Raes, Martine

    2016-03-23

    The outbreak of bovine spongiform encephalopathy (BSE) in the United Kingdom in 1986, with processed animal proteins (PAPs) as the main vector of the disease, has led to their prohibition in feed. The progressive release of the feed ban required the development of new analytical methods to determine the exact origin of PAPs from meat and bone meal. We set up a promising MS-based method to determine the species and the source (legal or not) present in PAPs: a TCA-acetone protein extraction followed by a cleanup step, an in-solution tryptic digestion of 5 h (with a 1:20 protein/trypsin ratio), and mass spectrometry analyses, first without any a priori, with a Q-TOF, followed by a targeted triple-quadrupole analysis. Using this procedure, we were able to overcome some of the major limitations of the official methods to analyze PAPs, detecting and identifying prohibited animal products in feedstuffs by the monitoring of peptides specific for cows, pigs, and sheep in PAPs. PMID:26943838

  3. Impact of Black seed (Nigella sativa) extract on bone turnover markers in postmenopausal women with osteoporosis

    OpenAIRE

    Valizadeh, N; H.R Zakeri; G Amin ansafi; A. Shafiee; Sarkhail, P.; Heshmat, R; H Sereshti; B Larijani

    2010-01-01

    "n "n  "n  "nBackground and the purpose of the study: "nExperimental studies have shown that Ns (Nigella sativa) seeds oil can increase bone formation and may have anabolic effects on bone loss. This study was conducted to investigate the beneficial impacts of the oil of Black seeds on bone turnover in osteoporotic postmenopausal women. "nMaterials and methods: A placebo controlled pilot study was carried out on 15 postmenopausal osteoporotic wo...

  4. Optimising the Use of TRIzol-extracted Proteins in Surface Enhanced Laser Desorption/ Ionization (SELDI Analysis

    Directory of Open Access Journals (Sweden)

    Perlaky Laszlo

    2006-03-01

    Full Text Available Abstract Background Research with clinical specimens is always hampered by the limited availability of relevant samples, necessitating the use of a single sample for multiple assays. TRIzol is a common reagent for RNA extraction, but DNA and protein fractions can also be used for other studies. However, little is known about using TRIzol-extracted proteins in proteomic research, partly because proteins extracted from TRIzol are very resistant to solubilization. Results To facilitate the use of TRIzol-extracted proteins, we first compared the ability of four different common solubilizing reagents to solubilize the TRIzol-extracted proteins from an osteosarcoma cell line, U2-OS. Then we analyzed the solubilized proteins by Surface Enhanced Laser Desorption/ Ionization technique (SELDI. The results showed that solubilization of TRIzol-extracted proteins with 9.5 M Urea and 2% CHAPS ([3-[(3-cholamidopropyl-dimethylammonio]propanesulfonate] (UREA-CHAPS was significantly better than the standard 1% SDS in terms of solubilization efficiency and the number of detectable ion peaks. Using three different types of SELDI arrays (CM10, H50, and IMAC-Cu, we demonstrated that peak detection with proteins solubilized by UREA-CHAPS was reproducible (r > 0.9. Further SELDI analysis indicated that the number of ion peaks detected in TRIzol-extracted proteins was comparable to a direct extraction method, suggesting many proteins still remain in the TRIzol protein fraction. Conclusion Our results suggest that UREA-CHAPS performed very well in solubilizing TRIzol-extracted proteins for SELDI applications. Protein fractions left over after TRIzol RNA extraction could be a valuable but neglected source for proteomic or biochemical analysis when additional samples are not available.

  5. Nucleic acid and protein extraction from electropermeabilized E. coli cells on a microfluidic chip

    OpenAIRE

    Matos, T.; Senkbeil, Silja; Mendonça, A; Queiroz, J. A.; Kutter, Jörg Peter; Bulow, L.

    2013-01-01

    Due to the extensive use of nucleic acid and protein analysis of bacterial samples, there is a need for simple and rapid extraction protocols for both plasmid DNA and RNA molecules as well as reporter proteins like the green fluorescent protein (GFP). In this report, an electropermeability technique has been developed which is based on exposing E. coli cells to low voltages to allow extraction of nucleic acids and proteins. The flow-through electropermeability chip used consists of a microflu...

  6. Anti-Cancer Effects of Protein Extracts from Calvatia lilacina, Pleurotus ostreatus and Volvariella volvacea

    Directory of Open Access Journals (Sweden)

    Jin-Yi Wu

    2011-01-01

    Full Text Available Calvatia lilacina (CL, Pleurotus ostreatus (PO and Volvariella volvacea (VV are widely distributed worldwide and commonly eaten as mushrooms. In this study, cell viabilities were evaluated for a human colorectal adenocarcinoma cell line (SW480 cells and a human monocytic leukemia cell line (THP-1 cells. Apoptotic mechanisms induced by the protein extracts of PO and VV were evaluated for SW480 cells. The viabilities of THP-1 and SW480 cells decreased in a concentration-dependent manner after 24 h of treatment with the protein extracts of CL, PO or VV. Apoptosis analysis revealed that the percentage of SW480 cells in the SubG1 phase (a marker of apoptosis was increased upon PO and VV protein-extract treatments, indicating that oligonucleosomal DNA fragmentation existed concomitantly with cellular death. The PO and VV protein extracts induced reactive oxygen species (ROS production, glutathione (GSH depletion and mitochondrial transmembrane potential (ΔΨm loss in SW480 cells. Pretreatment with N-acetylcysteine, GSH or cyclosporine A partially prevented the apoptosis induced by PO protein extracts, but not that induced by VV extracts, in SW480 cells. The protein extracts of CL, PO and VV exhibited therapeutic efficacy against human colorectal adenocarcinoma cells and human monocytic leukemia cells. The PO protein extracts induced apoptosis in SW480 cells partially through ROS production, GSH depletion and mitochondrial dysfunction. Therefore, the protein extracts of these mushrooms could be considered an important source of new anti-cancer drugs.

  7. Protein growth factors loaded highly porous chitosan scaffold: A comparison of bone healing properties

    Energy Technology Data Exchange (ETDEWEB)

    Nandi, Samit K., E-mail: samitnandi1967@gmail.com [Department of Veterinary Surgery and Radiology, West Bengal University of Animal and Fishery Sciences, Kolkata (India); Kundu, Biswanath, E-mail: biswa_kundu@rediffmail.com [Bioceramics and Coating Division, CSIR—Central Glass and Ceramic Research Institute, Kolkata (India); Basu, Debabrata [Bioceramics and Coating Division, CSIR—Central Glass and Ceramic Research Institute, Kolkata (India)

    2013-04-01

    Present study aimed to investigate and compare effectiveness of porous chitosan alone and in combination with insulin like growth factor-1 (IGF-1) and bone morphogenetic protein-2 (BMP-2) in bone healing. Highly porous (85 ± 2%) with wide distribution of macroporous (70–900 μm) chitosan scaffolds were fabricated as bone substitutes by employing a simple liquid hardening method using 2% (w/v) chitosan suspension. IGF-1 and BMP-2 were infiltrated using vacuum infiltration with freeze drying method. Adsorption efficiency was found to be 87 ± 2 and 90 ± 2% for BMP-2 and IGF-1 respectively. After thorough material characterization (pore details, FTIR and SEM), samples were used for subsequent in vivo animal trial. Eighteen rabbit models were used to evaluate and compare control (chitosan) (group A), chitosan with IGF-1 (group B) and chitosan with BMP-2 (group C) in the repair of critical size bone defect in tibia. Radiologically, there was evidence of radiodensity in defect area from 60th day (initiated on 30th day) in groups B and C as compared to group A and attaining nearly bony density in most of the part at day 90. Histological results depicted well developed osteoblastic proliferation around haversian canal along with proliferating fibroblast, vascularization and reticular network which was more pronounced in group B followed by groups C and A. Fluorochrome labeling and SEM studies in all groups showed similar outcome. Hence, porous chitosan alone and in combination with growth factors (GFs) can be successfully used for bone defect healing with slight advantage of IGF-1 in chitosan samples. - Highlights: ► Fabrication and characterization of porous chitosan with or without IGF-1 and BMP-2 ► Highly porous growth factor loaded chitosan studied in animal subjects for 3 months ► Parameters studied: histopathology, radiology and fluorochrome labeling ► IGF-1 loaded porous chitosan found to be very effective for bone defect healing.

  8. Protein growth factors loaded highly porous chitosan scaffold: A comparison of bone healing properties

    International Nuclear Information System (INIS)

    Present study aimed to investigate and compare effectiveness of porous chitosan alone and in combination with insulin like growth factor-1 (IGF-1) and bone morphogenetic protein-2 (BMP-2) in bone healing. Highly porous (85 ± 2%) with wide distribution of macroporous (70–900 μm) chitosan scaffolds were fabricated as bone substitutes by employing a simple liquid hardening method using 2% (w/v) chitosan suspension. IGF-1 and BMP-2 were infiltrated using vacuum infiltration with freeze drying method. Adsorption efficiency was found to be 87 ± 2 and 90 ± 2% for BMP-2 and IGF-1 respectively. After thorough material characterization (pore details, FTIR and SEM), samples were used for subsequent in vivo animal trial. Eighteen rabbit models were used to evaluate and compare control (chitosan) (group A), chitosan with IGF-1 (group B) and chitosan with BMP-2 (group C) in the repair of critical size bone defect in tibia. Radiologically, there was evidence of radiodensity in defect area from 60th day (initiated on 30th day) in groups B and C as compared to group A and attaining nearly bony density in most of the part at day 90. Histological results depicted well developed osteoblastic proliferation around haversian canal along with proliferating fibroblast, vascularization and reticular network which was more pronounced in group B followed by groups C and A. Fluorochrome labeling and SEM studies in all groups showed similar outcome. Hence, porous chitosan alone and in combination with growth factors (GFs) can be successfully used for bone defect healing with slight advantage of IGF-1 in chitosan samples. - Highlights: ► Fabrication and characterization of porous chitosan with or without IGF-1 and BMP-2 ► Highly porous growth factor loaded chitosan studied in animal subjects for 3 months ► Parameters studied: histopathology, radiology and fluorochrome labeling ► IGF-1 loaded porous chitosan found to be very effective for bone defect healing

  9. Deer bone extract suppresses articular cartilage damage induced by monosodium iodoacetate in osteoarthritic rats: an in vivo micro-computed tomography study.

    Science.gov (United States)

    Lee, Hyunji; Park, Yooheon; Ahn, Chang Won; Park, Soo Hyun; Jung, Eun Young; Suh, Hyung Joo

    2014-06-01

    We evaluated the anti-osteoarthritic effects of deer bone extract on articular cartilage damage by using micro-computed tomography (micro-CT) in monosodium iodoacetate (MIA)-induced osteoarthritis (OA) in rats. Male Wistar rats (6 weeks of age) were randomly divided into 5 groups (10 rats/group): sham control (SC; PBS injection+PBS 1 mL treatment); negative control (NC; MIA injection+PBS 1 mL treatment); positive control (PC; MIA injection+250 mg/kg glucosamine sulfate/chondroitin sulfate mixture treatment); low dose (LDB; MIA injection+250 mg/kg deer bone extract treatment); and high dose (HDB; MIA injection+500 mg/kg deer bone extract treatment). After 50 days of treatment, we observed that the administration of deer bone extract protected against bone destruction and reduced the number of erosion lacunae. When deer bone extract was administered, the trabecular thickness distribution (Tb.Th) (LDB: 75.9 μm, HDB: 80.7 μm vs. NC: 48.0 μm) and the trabecular bone volume fraction ratio (BV/TV) (LDB: 43.8%, HDB: 48.2% vs. NC: 39.1%) were significantly restored. Additionally, the trabecular separation (Tb.Sp) increase caused by MIA was decreased significantly with the administration of deer bone extract (LDB: 73.4 μm, HDB: 81.2 μm vs. NC: 112.0 μm). We concluded that the oral administration of deer bone extract effectively relieved the morphological changes induced by MIA injection in an animal model. PMID:24797662

  10. Effect of chickpea aqueous extracts, organic extracts, and protein concentrates on cell proliferation.

    Science.gov (United States)

    Girón-Calle, Julio; Vioque, Javier; del Mar Yust, María; Pedroche, Justo; Alaiz, Manuel; Millán, Francisco

    2004-01-01

    Pulses should be part of a healthy diet, and it is also becoming clear that they have health-promoting effects. Nevertheless, most studies on the bioactive or health-promoting properties of pulses have been carried out using soybeans. We have studied cell growth-regulating properties, which may be responsible for anti-cancer properties, in chickpea seeds. Chickpea seeds are a staple in the traditional diet of many Mediterranean, Asian, and South and Central American countries. In addition, chickpea seeds have industrial applications since they can be used for the preparation of protein concentrates and isolates. The cell lines Caco-2 (epithelial intestinal) and J774 (macrophages) have been exposed to chickpea seed extracts and protein preparations in order to screen the different chickpea fractions for effects on cell growth. Both cell growth-promoting and cell growth-inhibiting effects were found. Most interestingly, a fraction soluble in ethanol and acetone specifically and almost completely inhibited the growth of Caco-2 cells exhibiting a cancerous phenotype. It is concluded that chickpea seeds are a source of bioactive components and deserve further study for their possible anti-cancer effect. PMID:15298756

  11. The application of bone morphogenetic proteins to periodontal and peri-implant tissue regeneration: A literature review

    OpenAIRE

    Sasikumar, Karuppanan P.; Sugumari Elavarasu; Jayaprakash S Gadagi

    2012-01-01

    Progress in understanding the role of bone morphogenetic proteins (BMPs) in craniofacial and tooth development and the demonstration of stem cells in periodontal ligament have set the stage for periodontal regenerative therapy and tissue engineering. Furthermore, recent approval by the Food and Drug Administration of recombinant human BMPs for accelerating bone fusion in slow-healing fractures indicates that this protein family may prove useful in designing regenerative treatments in periodon...

  12. Glucocorticoid-induced differentiation of fetal rat calvarial osteoblasts is mediated by bone morphogenetic protein-6.

    Science.gov (United States)

    Boden, S D; Hair, G; Titus, L; Racine, M; McCuaig, K; Wozney, J M; Nanes, M S

    1997-07-01

    Glucocorticoids (GCs) at physiological concentrations promote osteoblast differentiation from fetal calvarial cells, calvarial organ cultures, and bone marrow stromal cells; however, the cellular pathways involved are not known. Bone morphogenetic proteins (BMPs) are recognized as important mediators of osteoblast differentiation. Specific roles for individual BMPs during postembryonic membranous bone formation have yet to be determined. We recently reported that GC potentiated the osteoblast differentiation effects of BMP-2 and BMP-4, but not of BMP-6, which, by itself, was the most potent of the three. In the present study, we used fetal rat secondary calvarial cultures to study the role of BMP-6 during early osteoblast differentiation. Treatment with the GC triamcinolone (10(-9) M) resulted in a 5- to 8-fold increase in BMP-6 steady-state messenger RNA levels, peaking at 12 h. In contrast, BMPs -2, -4, -5, -7, and transforming growth factor (TGF)-beta1 messenger RNA levels increased by less than 2-fold, after GC treatment, compared with untreated control cultures at 24 h. BMP-6 protein secretion increased 6- to 7-fold by 12 h and 12-fold (from 7.5 to 90 ng/ml) by 24 h, as measured by quantitative Western analysis. Treatment of cells with oligodeoxynucleotides antisense to BMP-6 diminished secretion of BMP-6 protein and significantly inhibited the GC-induced differentiation, as determined by a 10-fold decrease in the number of mineralized bone nodules, compared with controls that were treated with sense oligonucleotides or no oligonucleotides (ANOVA, P < 0.05). The antisense oligonucleotide inhibition of differentiation was rescued by treatment with exogenous recombinant human BMP-6. We conclude that GC-induced differentiation of osteoblasts from the pluripotent precursors is mediated, in part, by BMP-6. These results suggest that BMP-6 has an important and unique role during early osteoblast differentiation. PMID:9202223

  13. Nucleic Acids and Protein Metabolism of Bone Marrow Cells Studied by Means of Tritiumlabelled Precursors

    International Nuclear Information System (INIS)

    The advantages of the use of tritium-labelled compounds in radioautographic technique are discussed. Tritium electrons have a maximal energy of 0.018 MeV, corresponding to about 1μm range in a photographic emulsion, and consequently they allow the highest possible resolution at a cellular and subcellular level. This is particularly useful for studying metabolic phenomena of tissues which are composed, as in the case of bone marrow, of different cellular types at various stages of differentiation. This technique has been used for investigating nucleic acids and protein metabolism of normal and leukaemic bone marrow cells. DNA metabolism has been studied utilizing a specific precursor, H3-thymidine. Some significant differences of the percentages of labelled cells have been detected by comparing the normal and leukaemic elements belonging to the same stage of maturation. In acute leukaemia cells, particularly, a strikingly lower incorporation of thymidine was found and these results have been taken as evidence of a decreased proliferative capacity of these cells, as compared to normal myeloblasts. With the same technique, RNA and protein metabolism have been investigated utilizing H3- uridine, H3-leucine and H3-phenylalanine as precursors. The existence of a strict interrelationship between RNA and protein metabolism is now fully accepted in cellular biology. The existence of a constant ratio between uridine and amino acids incorporation has also been demonstrated in normal bone marrow cells. In acute leukaemia cells the incorporation of RNA and protein precursors, although different from case to case, is constantly and significantly lower. Furthermore, the ratio between uridine and amino acids incorporation is constantly altered in these cells. The lower RNA and protein metabolism and its dissociation in acute leukaemia cells is discussed in relation to the well-known maturation defect of these cells. (author)

  14. The Effectiveness of Crataegus orientalis M Bieber. (Hawthorn) Extract Administration in Preventing Alveolar Bone Loss in Rats with Experimental Periodontitis.

    Science.gov (United States)

    Hatipoğlu, Mükerrem; Sağlam, Mehmet; Köseoğlu, Serhat; Köksal, Ekrem; Keleş, Ali; Esen, Hacı Hasan

    2015-01-01

    The purpose of this animal study was to evaluate the effects of hawthorn (Crataeus orientalis M Bieber.) extract on serum oxidative status and alveolar bone loss in experimental periodontitis. Twenty-seven Wistar rats were assigned to one of the following groups: non- ligated+placebo (saline) (NL, n = 9), ligature only+placebo (saline) (LO, n = 9), and ligature and treated with hawthorn extract in saline (H, n = 9) (100 mg/kg orogastrically, once a day for 11 days). Periodontitis was induced by submerging a 4/0 silk ligature in the sulcus of the mandibular right first molars of rats, and the animals were sacrificed after 11 days. Micro-CT examinations were performed for linear and volumetric parameter assessment of alveolar bone. Periodontal tissues were histopathologically examined to assess the differences among the study groups. Levels of serum total antioxidant status (TAS)/total oxidant status (TOS), and oxidative stress index (OSI) were also analyzed. Alveolar bone loss was significantly reduced by hawthorn administration compared to LO group (p<0.05). The number of inflammatory cells and osteoclasts in the LO group was significantly higher than that of the NL and H groups (p< 0.05). The number of osteoblasts in the LO and H groups was significantly higher than that of the NL group (p<0.05). TOS and OSI levels were significantly reduced in H group compared to LO group (P <0.05) and TAS levels were similar in H and NL group (p< 0.05). Hawthorn extract showed inhibitory effect on periodontal inflammation and alveolar bone loss by regulating TAS, TOS and OSI levels in periodontal disease in rats when administered systemically. PMID:26030160

  15. A new biocompatible delivery scaffold containing heparin and bone morphogenetic protein 2

    Directory of Open Access Journals (Sweden)

    Thanyaphoo Suphannee

    2016-09-01

    Full Text Available Silicon-substituted calcium phosphate (Si-CaP was developed in our laboratory as a biomaterial for delivery in bone tissue engineering. It was fabricated as a 3D-construct of scaffolds using chitosan-trisodium polyphosphate (TPP cross-linked networks. In this study, heparin was covalently bonded to the residual -NH2 groups of chitosan on the scaffold applying carbodiimide chemistry. Bonded heparin was not leached away from scaffold surfaces upon vigorous washing or extended storage. Recombinant human bone morphogenetic protein 2 (rhBMP-2 was bound to conjugated scaffolds by ionic interactions between the negatively charged SO42- clusters of heparin and positively charged amino acids of rhBMP-2. The resulting scaffolds were inspected for bone regenerative capacity by subcutaneous implanting in rats. Histological observation and mineralization assay were performed after 4 weeks of implantation. Results from both in vitro and in vivo experiments suggest the potential of the developed scaffolds for bone tissue engineering applications in the future.

  16. A new biocompatible delivery scaffold containing heparin and bone morphogenetic protein 2.

    Science.gov (United States)

    Thanyaphoo, Suphannee; Kaewsrichan, Jasadee

    2016-09-01

    Silicon-substituted calcium phosphate (Si-CaP) was developed in our laboratory as a biomaterial for delivery in bone tissue engineering. It was fabricated as a 3D-construct of scaffolds using chitosan-trisodium polyphosphate (TPP) cross-linked networks. In this study, heparin was covalently bonded to the residual -NH2 groups of chitosan on the scaffold applying carbodiimide chemistry. Bonded heparin was not leached away from scaffold surfaces upon vigorous washing or extended storage. Recombinant human bone morphogenetic protein 2 (rhBMP-2) was bound to conjugated scaffolds by ionic interactions between the negatively charged SO42- clusters of heparin and positively charged amino acids of rhBMP-2. The resulting scaffolds were inspected for bone regenerative capacity by subcutaneous implanting in rats. Histological observation and mineralization assay were performed after 4 weeks of implantation. Results from both in vitro and in vivo experiments suggest the potential of the developed scaffolds for bone tissue engineering applications in the future. PMID:27383886

  17. Characterization of rice starch and protein obtained by a fast alkaline extraction method.

    Science.gov (United States)

    Souza, Daiana de; Sbardelotto, Arthur Francisco; Ziegler, Denize Righetto; Marczak, Ligia Damasceno Ferreira; Tessaro, Isabel Cristina

    2016-01-15

    This study evaluated the characteristics of rice starch and protein obtained by a fast alkaline extraction method on rice flour (RF) derived from broken rice. The extraction was conducted using 0.18% NaOH at 30°C for 30min followed by centrifugation to separate the starch rich and the protein rich fractions. This fast extraction method allowed to obtain an isoelectric precipitation protein concentrate (IPPC) with 79% protein and a starchy product with low protein content. The amino acid content of IPPC was practically unchanged compared to the protein in RF. The proteins of the IPPC underwent denaturation during extraction and some of the starch suffered the cold gelatinization phenomenon, due to the alkaline treatment. With some modifications, the fast method can be interesting in a technological point of view as it enables process cost reduction and useful ingredients obtention to the food and chemical industries. PMID:26258699

  18. Ameloblastin, an Extracellular Matrix Protein, Affects Long Bone Growth and Mineralization.

    Science.gov (United States)

    Lu, Xuanyu; Fukumoto, Satoshi; Yamada, Yoshihiko; Evans, Carla A; Diekwisch, Thomas Gh; Luan, Xianghong

    2016-06-01

    Matrix molecules such as the enamel-related calcium-binding phosphoprotein ameloblastin (AMBN) are expressed in multiple tissues, including teeth, bones, and cartilage. Here we have asked whether AMBN is of functional importance for timely long bone development and, if so, how it exerts its function related to osteogenesis. Adolescent AMBN-deficient mice (AMBN(Δ5-6) ) suffered from a 33% to 38% reduction in femur length and an 8.4% shorter trunk spinal column when compared with WT controls, whereas there was no difference between adult animals. On a cellular level, AMBN truncation resulted in a shortened growth plate and a 41% to 49% reduction in the number of proliferating tibia chondrocytes and osteoblasts. Bone marrow stromal cells (BMSCs) isolated from AMBN mutant mice displayed defects in proliferation and differentiation potential as well as cytoskeleton organization. Osteogenesis-related growth factors, such as insulin-like growth factor 1 (IGF1) and BMP7, were also significantly (46% to 73%) reduced in AMBN-deficient BMSCs. Addition of exogenous AMBN restored cytoskeleton structures in AMBN mutant BMSCs and resulted in a dramatic 400% to 600% increase in BMP2, BMP7, and Col1A expression. Block of RhoA diminished the effect of AMBN on osteogenic growth factor and matrix protein gene expression. Addition of exogenous BMP7 and IGF1 rescued the proliferation and differentiation potential of AMBN-deficient BMSCs. Confirming the effects of AMBN on long bone growth, back-crossing of mutant mice with full-length AMBN overexpressors resulted in a complete rescue of AMBN(Δ5-6) bone defects. Together, these data indicate that AMBN affects extracellular matrix production and cell adhesion properties in the long bone growth plate, resulting in altered cytoskeletal dynamics, increased osteogenesis-related gene expression, as well as osteoblast and chondrocyte proliferation. We propose that AMBN facilitates rapid long bone growth and an important growth spurt during the

  19. Cardiovascular Calcification and Bone : A Comparison of the Effects of Dietary and Serum Vitamin K and its Dependent Proteins

    OpenAIRE

    Nicoll, Rachel; McLaren Howard, John; Michael Y. Henein

    2015-01-01

    This review compares the effect of vitamin K on cardiovascular (CV) calcification and bone health and shows that, in principal, the γ-carboxylation of the vitamin K-dependent proteins matrix Gla protein (MGP) and its bone equivalent osteocalcin (OC), generally ensures that hydroxyapatite is kept out of the CV system and is deposited in bone. This is an important finding, since there is currently no reliable treatment for CV calcification.Vitamin K2 (menaquinone) may be more effective in the a...

  20. Comparison of Three Methods of Protein Extraction from Dermatophagoides Pteronyssinus for Two-dimensional Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Jin-lu Sun; Hong-yu Zhang; Zhi-yi Guo; Wan-tao Ying; Xiao-hong Qian; Jing-lan Wang

    2009-01-01

    Objective To explore an effective method of Dermatophagoides pteronyssinus protein extraction suitable for two-dimensional electrophoresis (2-DE) analysis. Methods The extracts of Dermatophagoides pteronyssinus were prepared with Coca's solution, lysis buffer of 2-DE, and Trizol reagent, respectively. Bicinchoninic acid (BCA) assay was used to determine the total protein concentration of the samples. The efficiency of different protein extraction methods were evaluated with 2-DE analysis. Results The concentrations of extracted protein by methods of Coca's solution, lysis buffer, and Trizol reagent were 0.63 g/L, 0.90 g/L, and 0.80 g/L, respectively. The 2-DE analysis results showed that some protein spots in low molecular weight (LMW) range could be detected with the Coca's solution method. With the lysis buffer of 2-DE method, more protein spots in LMW range could be detected, while the medium molecular weight (MMW) protein spots were absent. Several MMW protein spots (174-178 kD and 133 kD) and more LMW protein spots were detected with Trizol reagent method. Conclusions Among Coca's solution, lysis buffer of 2-DE, and Trizol reagent, the concentration of extracted protein of Dermatophagoides pteronyssinus by lysis buffer of 2-DE is the highest. However, most protein components of Dermatophagoides pteronyssinus purified mite bodies can be extracted by Trizol reagent, which may generally reflect the whole profile of Dermatophagoides pteronyssinus allergens.

  1. Advance chromatin extraction improves capture performance of protein A affinity chromatography.

    Science.gov (United States)

    Nian, Rui; Zhang, Wei; Tan, Lihan; Lee, Jeremy; Bi, Xeuzhi; Yang, Yuansheng; Gan, Hui Theng; Gagnon, Pete

    2016-01-29

    Practical effects of advance chromatin removal on performance of protein A affinity chromatography were evaluated using a caprylic acid-allantoin-based extraction method. Lacking this treatment, the practice of increasing loading residence time to increase capacity was shown to increase host protein contamination of the eluted IgG. Advance chromatin extraction suspended that compromise. Protein A ligand leakage from columns loaded with chromatin-extracted harvest was half the level observed on protein A columns loaded with non-extracted harvest. Columns loaded with chromatin-extracted harvest were cleaned more effectively by 50-100mM NaOH than columns loaded with non-extracted harvest that were cleaned with 250-500mM NaOH. Two protein A media with IgG capacities in excess of 50g/L were loaded with chromatin-extracted harvest, washed with 2.0M NaCl before elution, and the eluted IgG fraction titrated to pH 5.5 before microfiltration. Host protein contamination in the filtrate was reduced to <1ppm, DNA to <1ppb, protein A leakage to 0.5ppm, and aggregates to 1.0%. Caprylic acid and allantoin were both reduced below 5ppm. Step recovery of IgG was 99.4%. Addition of a single polishing step reduced residual protein A beneath the level of detection and aggregates to <0.1%. Overall process recovery including chromatin extraction was 90%. PMID:26774119

  2. Extraction of up to 95% of unreduced wheat flour protein using warm SDS

    Science.gov (United States)

    Extraction of intact glutenin polymer without sonication is an essential prerequisite for accurate determination of the molecular size distribution of the polymers. Sequential fractionation of wheat flour extracted up to 95% of total protein and up to 92% of polymeric protein without sonication or r...

  3. Slow acting protein extract from fruit pulp of Momordica charantia with insulin secretagogue and insulinomimetic activities.

    Science.gov (United States)

    Yibchok-anun, Sirintorn; Adisakwattana, Sirichai; Yao, Cheng Yu; Sangvanich, Polkit; Roengsumran, Sophon; Hsu, Walter Haw

    2006-06-01

    The protein from Thai bitter gourd (Momordica charantia) fruit pulp was extracted and studied for its hypoglycemic effect. Subcutaneous administration of the protein extract (5, 10 mg/kg) significantly and markedly decreased plasma glucose concentrations in both normal and streptozotocin-induced diabetic rats in a dose-dependent manner. The onset of the protein extract-induced antihyperglycemia/hypoglycemia was observed at 4 and 6 h in diabetic and normal rats, respectively. This protein extract also raised plasma insulin concentrations by 2 fold 4 h following subcutaneous administration. In perfused rat pancreas, the protein extract (10 microg/ml) increased insulin secretion, but not glucagon secretion. The increase in insulin secretion was apparent within 5 min of administration and was persistent during 30 min of administration. Furthermore, the protein extract enhanced glucose uptake into C2C12 myocytes and 3T3-L1 adipocytes. Time course experiments performed in rat adipocytes revealed that M. charantia protein extract significantly increased glucose uptake after 4 and 6 h of incubation. Thus, the M. charantia protein extract, a slow acting chemical, exerted both insulin secretagogue and insulinomimetic activities to lower blood glucose concentrations in vivo. PMID:16755004

  4. Murine Hind Limb Long Bone Dissection and Bone Marrow Isolation.

    Science.gov (United States)

    Amend, Sarah R; Valkenburg, Kenneth C; Pienta, Kenneth J

    2016-01-01

    Investigation of the bone and the bone marrow is critical in many research fields including basic bone biology, immunology, hematology, cancer metastasis, biomechanics, and stem cell biology. Despite the importance of the bone in healthy and pathologic states, however, it is a largely under-researched organ due to lack of specialized knowledge of bone dissection and bone marrow isolation. Mice are a common model organism to study effects on bone and bone marrow, necessitating a standardized and efficient method for long bone dissection and bone marrow isolation for processing of large experimental cohorts. We describe a straightforward dissection procedure for the removal of the femur and tibia that is suitable for downstream applications, including but not limited to histomorphologic analysis and strength testing. In addition, we outline a rapid procedure for isolation of bone marrow from the long bones via centrifugation with limited handling time, ideal for cell sorting, primary cell culture, or DNA, RNA, and protein extraction. The protocol is streamlined for rapid processing of samples to limit experimental error, and is standardized to minimize user-to-user variability. PMID:27168390

  5. Characterisation of kiwifruit and asparagus enzyme extracts, and their activities toward meat proteins.

    Science.gov (United States)

    Ha, Minh; Bekhit, Alaa El-Din; Carne, Alan; Hopkins, David L

    2013-01-15

    Two plant enzyme extracts from kiwifruit and asparagus were evaluated for their ability to hydrolyse commercially available substrates and proteins present in both beef connective tissue and topside myofibrillar extracts. The results show significant differences in protease activity depending on the assay used. Protease assays with connective tissue and meat myofibrillar extracts provide a more realistic evaluation of the potential of the enzymes for application in meat tenderization. Overall, the kiwifruit protease extract was found to be more effective at hydrolysing myofibrillar and collagen proteins than the asparagus protease extract. The two protease extracts appeared to target meat myofibrillar and collagen proteins differently, suggesting the potential of a synergistic effect of these proteases in improving the tenderness of specific cuts of meat, based on their intrinsic protein composition. PMID:23122154

  6. Effects of Osseointegration by Bone Morphogenetic Protein-2 on Titanium Implants In Vitro and In Vivo.

    Science.gov (United States)

    Teng, Fu-Yuan; Chen, Wen-Cheng; Wang, Yin-Lai; Hung, Chun-Cheng; Tseng, Chun-Chieh

    2016-01-01

    This study designed a biomimetic implant for reducing healing time and achieving early osseointegration to create an active surface. Bone morphogenetic protein-2 (BMP-2) is a strong regulator protein in osteogenic pathways. Due to hardly maintaining BMP-2 biological function and specificity, BMP-2 efficient delivery on implant surfaces is the main challenge for the clinic application. In this study, a novel method for synthesizing functionalized silane film for superior modification with BMP-2 on titanium surfaces is proposed. Three groups were compared with and without BMP-2 on modified titanium surfaces in vitro and in vivo: mechanical grinding; electrochemical modification through potentiostatic anodization (ECH); and sandblasting, alkali heating, and etching (SMART). Cell tests indicated that the ECH and SMART groups with BMP-2 markedly promoted D1 cell activity and differentiation compared with the groups without BMP-2. Moreover, the SMART group with a BMP-2 surface markedly promoted early alkaline phosphatase expression in the D1 cells compared with the other surface groups. Compared with these groups in vivo, SMART silaning with BMP-2 showed superior bone quality and created contact areas between implant and surrounding bones. The SMART group with BMP-2 could promote cell mineralization in vitro and osseointegration in vivo, indicating potential clinical use. PMID:26977141

  7. Effects of Osseointegration by Bone Morphogenetic Protein-2 on Titanium Implants In Vitro and In Vivo

    Directory of Open Access Journals (Sweden)

    Fu-Yuan Teng

    2016-01-01

    Full Text Available This study designed a biomimetic implant for reducing healing time and achieving early osseointegration to create an active surface. Bone morphogenetic protein-2 (BMP-2 is a strong regulator protein in osteogenic pathways. Due to hardly maintaining BMP-2 biological function and specificity, BMP-2 efficient delivery on implant surfaces is the main challenge for the clinic application. In this study, a novel method for synthesizing functionalized silane film for superior modification with BMP-2 on titanium surfaces is proposed. Three groups were compared with and without BMP-2 on modified titanium surfaces in vitro and in vivo: mechanical grinding; electrochemical modification through potentiostatic anodization (ECH; and sandblasting, alkali heating, and etching (SMART. Cell tests indicated that the ECH and SMART groups with BMP-2 markedly promoted D1 cell activity and differentiation compared with the groups without BMP-2. Moreover, the SMART group with a BMP-2 surface markedly promoted early alkaline phosphatase expression in the D1 cells compared with the other surface groups. Compared with these groups in vivo, SMART silaning with BMP-2 showed superior bone quality and created contact areas between implant and surrounding bones. The SMART group with BMP-2 could promote cell mineralization in vitro and osseointegration in vivo, indicating potential clinical use.

  8. Nanoscale control of silica particle formation via silk-silica fusion proteins for bone regeneration.

    Science.gov (United States)

    Mieszawska, Aneta J; Nadkarni, Lauren D; Perry, Carole C; Kaplan, David L

    2010-10-26

    The biomimetic design of silk/silica fusion proteins was carried out, combining the self assembling domains of spider dragline silk (Nephila clavipes) and silaffin derived R5 peptide of Cylindrotheca fusiformis that is responsible for silica mineralization. Genetic engineering was used to generate the protein-based biomaterials incorporating the physical properties of both components. With genetic control over the nanodomain sizes and chemistry, as well as modification of synthetic conditions for silica formation, controlled mineralized silk films with different silica morphologies and distributions were successfully generated; generating 3D porous networks, clustered silica nanoparticles (SNPs), or single SNPs. Silk serves as the organic scaffolding to control the material stability and multiprocessing makes silk/silica biomaterials suitable for different tissue regenerative applications. The influence of these new silk-silica composite systems on osteogenesis was evaluated with human mesenchymal stem cells (hMSCs) subjected to osteogenic differentiation. hMSCs adhered, proliferated, and differentiated towards osteogenic lineages on the silk/silica films. The presence of the silica in the silk films influenced osteogenic gene expression, with the upregulation of alkaline phosphatase (ALP), bone sialoprotein (BSP), and collagen type 1 (Col 1) markers. Evidence for early bone formation as calcium deposits was observed on silk films with silica. These results indicate the potential utility of these new silk/silica systems towards bone regeneration. PMID:20976116

  9. Influence of bone morphogenetic protein type IA receptor conditional knockout in lens on expression of bone morphogenetic protein 4 in lens

    Institute of Scientific and Technical Information of China (English)

    Qi; Zhao; Jiang-Yue; Zhao; Jin-Song; Zhang

    2015-01-01

    AIM: To investigate the influence of bone morphogenetic protein type IA receptor [BMPR-IA(ALK3)] conditional knockout in lens on expression of bone morphogenetic protein 4(BMP4) in lens during the development of the vertebrate eye.METHODS: Cre-positive mice were mated with Crenegative mice to generate 50% Cre-positive(conditional knockout, CKO) 4 embryos, 8 eyes and 50% Cre-negative offspring(wild type, WT) 4 embryos, 8 eyes. The embryos were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned to a thickness of 4 μm.Removal of paraffin wax and dehydrating for sections,and then the procedure of in situ hybridization was processed, BMP4 MK1784-m(BOSTER) was used, and observed the expression of BMP4 in the lens in experimental group and control group. We selected SPSS11.0 software for statistical analysis, P<0.05 showed that the difference was statistically significant.· RESULTS: Four embryos of each genotype were examined, totally we had 8 embryos, 16 eyes. We got the uniform outcomes in all the embryos. We found ALK3 was required during lens growing, but was not essential for the formation of lens. We observed that the expression of BMP4 in the lens was significantly reduced in all 8 ALK3 CKO lens, BMP4 expression was normal in all the 8 WT lens, P <0.01. This phenomenon became increasingly visible in accordance with embryo development. The most apparent alteration was present at stage E15.5.CONCLUSION: ALK3 is essential for lens growth. The influence of ALK3 on the expression of BMP4 is present during the development of mice lens.

  10. Effects of magnesium alloys extracts on adult human bone marrow-derived stromal cell viability and osteogenic differentiation

    Energy Technology Data Exchange (ETDEWEB)

    Yang Chunxi; Dai Kerong [Department of Orthopedics, Ninth People' s Hospital, Shanghai Jiao Tong University School of Medicine, 639 Zhizaoju Road, Shanghai 200011 (China); Yuan Guangyin; Zhang Jia [National Engineering Research Center of Light Alloys Net Forming (LAF), School of Materials Science and Engineering, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240 (China); Tang Ze; Zhang Xiaoling [Lab of Osteopaedic Cellular and Molecular Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences (SIBS), Chinese Academy of Sciences (CAS) and Shanghai Jiao Tong University School of Medicine - SJTUSM, Shanghai 200025 (China)

    2010-08-01

    In this study, adult human bone marrow-derived stromal cells (hBMSCs) were cultured in extracts of magnesium (Mg) and the Mg alloys AZ91D and NZ30K for 12 days. We studied the indirect effects of Mg alloys on hBMSC viability. Alkaline phosphatase activity and the expression of osteogenic differentiation marker genes were used to evaluate the effects of the Mg alloys on the osteogenic differentiation of hBMSCs. The results indicate that {<=}10 mM concentration of Mg in the extracts did not inhibit the viability and osteogenic differentiation of hBMSCs. However, the results suggest that the high pH of the extracts, which is a result of the rapid corrosion of Mg and the Mg alloys, is unfavorable to the viability and osteogenic differentiation of hBMSCs.

  11. Effects of magnesium alloys extracts on adult human bone marrow-derived stromal cell viability and osteogenic differentiation

    International Nuclear Information System (INIS)

    In this study, adult human bone marrow-derived stromal cells (hBMSCs) were cultured in extracts of magnesium (Mg) and the Mg alloys AZ91D and NZ30K for 12 days. We studied the indirect effects of Mg alloys on hBMSC viability. Alkaline phosphatase activity and the expression of osteogenic differentiation marker genes were used to evaluate the effects of the Mg alloys on the osteogenic differentiation of hBMSCs. The results indicate that ≤10 mM concentration of Mg in the extracts did not inhibit the viability and osteogenic differentiation of hBMSCs. However, the results suggest that the high pH of the extracts, which is a result of the rapid corrosion of Mg and the Mg alloys, is unfavorable to the viability and osteogenic differentiation of hBMSCs.

  12. Starch extraction process coupled to protein recovery from leguminous tuberous roots (Pachyrhizus ahipa).

    Science.gov (United States)

    Díaz, Andrea; Dini, Cecilia; Viña, Sonia Z; García, María A

    2016-11-01

    The objective of this work was to fit together the starch extraction from Pachyrhizus ahipa roots and the recovery of the proteins present in these storage organs, making an improved use of this novel raw material. The replacement of water by buffer PO4(-3)/NaCl as solvent in the first extraction steps improved protein extraction without lowering the starch yield. The starches obtained from the traditional and the proposed methods exhibited some differences in appearance and technological and thermal properties, which were endorsed to the adjustment in the methodology of extraction rather than to the use of buffer as solvent. Thus, P. ahipa starch obtaining procedure could be coupled to protein extraction with a minimum change in the methodology. This innovation did not significantly shift the characteristics of the starch obtained and allowed to obtain a protein yield of 135.7mg BSA equivalent protein/100g of fresh roots. PMID:27516269

  13. The Mitigating Effect of Citrullus colocynthis (L. Fruit Extract against Genotoxicity Induced by Cyclophosphamide in Mice Bone Marrow Cells

    Directory of Open Access Journals (Sweden)

    Mohammad Shokrzadeh

    2013-01-01

    Full Text Available Possible genoprotective effect of Citrullus colocynthis (L. (CCT fruits extract against cyclophosphamide- (CP-induced DNA damage in mice bone marrow cells was evaluated using micronucleus assay, as an index of induced chromosomal damage. Mice were preadministered with different doses of CCT via intraperitoneal injection for 7 consecutive days followed by injection with CP (70 mg/kg b.w. 1 hr after the last injection of CCT. After 24 hr, mice were scarified to evaluate the frequency of micronucleated polychromatic erythrocytes (MnPCEs. In addition, the number of polychromatic erythrocytes (PCEs among 1000 normochromatic erythrocytes (NCEs per animal was recorded to evaluate bone marrow. Pretreatment with CCT significantly reduced the number of MnPCEs induced by CP in bone marrow cells (P<0.0001. At 200 mg/kg, CCT had a maximum chemoprotective effect and reduced the number of MnPCEs by 6.37-fold and completely normalized the mitotic activity. CCT also led to marked proliferation and hypercellularity of immature myeloid elements after mice were treated with CP and mitigated the bone marrow suppression. Our study revealed that CCT has an antigenotoxic effect against CP-induced oxidative DNA damage in mice. Therefore, it could be used concomitantly as a supplement to protect people undergoing chemotherapy.

  14. Inhibitory effects of French pine bark extract, Pycnogenol®, on alveolar bone resorption and on the osteoclast differentiation.

    Science.gov (United States)

    Sugimoto, Hideki; Watanabe, Kiyoko; Toyama, Toshizo; Takahashi, Shun-suke; Sugiyama, Shuta; Lee, Masaichi-Chang-il; Hamada, Nobushiro

    2015-02-01

    Pycnogenol(®) (PYC) is a standardized bark extract from French maritime pine (Pinus pinaster Aiton). We examined the inhibitory effects of PYC on alveolar bone resorption, which is a characteristic feature of periodontitis, induced by Porphyromonas gingivalis (P. gingivalis) and osteoclast differentiation. In rat periodontitis model, rats were divided into four groups: group A served as the non-infected control, group B was infected orally with P. gingivalis ATCC 33277, group C was administered PYC in the diet (0.025%: w/w), and group D was infected with P. gingivalis and administered PYC. Administration of PYC along with P. gingivalis infection significantly reduced alveolar bone resorption. Treatment of P. gingivalis with 1 µg/ml PYC reduced the number of viable bacterial cells. Addition of PYC to epithelial cells inhibited adhesion and invasion by P. gingivalis. The effect of PYC on osteoclast formation was confirmed by tartrate-resistant acid phosphatase staining. PYC treatment significantly inhibited osteoclast formation. Addition of PYC (1-100 µg/ml) to purified osteoclasts culture induced cell apoptosis. These results suggest that PYC may prevent alveolar bone resorption through its antibacterial activity against P. gingivalis and by suppressing osteoclastogenesis. Therefore, PYC may be useful as a therapeutic and preventative agent for bone diseases such as periodontitis. PMID:25336411

  15. An intramembranous ossification model for the in silico analysis of bone tissue formation in tooth extraction sites.

    Science.gov (United States)

    Corredor-Gómez, Jennifer Paola; Rueda-Ramírez, Andrés Mauricio; Gamboa-Márquez, Miguel Alejandro; Torres-Rodríguez, Carolina; Cortés-Rodríguez, Carlos Julio

    2016-07-21

    The accurate modeling of biological processes allows us to predict the spatiotemporal behavior of living tissues by computer-aided (in silico) testing, a useful tool for the development of medical strategies, avoiding the expenses and potential ethical implications of in vivo experimentation. A model for bone healing in mouth would be useful for selecting proper surgical techniques in dental procedures. In this paper, the formulation and implementation of a model for Intramembranous Ossification is presented aiming to describe the complex process of bone tissue formation in tooth extraction sites. The model consists in a mathematical description of the mechanisms in which different types of cells interact, synthesize and degrade extracellular matrices under the influence of biochemical factors. Special attention is given to angiogenesis, oxygen-dependent effects and growth factor-induced apoptosis of fibroblasts. Furthermore, considering the depth-dependent vascularization of mandibular bone and its influence on bone healing, a functional description of the cell distribution on the severed periodontal ligament (PDL) is proposed. The developed model was implemented using the finite element method (FEM) and successfully validated by simulating an animal in vivo experiment on dogs reported in the literature. A good fit between model outcome and experimental data was obtained with a mean absolute error of 3.04%. The mathematical framework presented here may represent an important tool for the design of future in vitro and in vivo tests, as well as a precedent for future in silico studies on osseointegration and mechanobiology. PMID:27113783

  16. Radiobiological long-term accumulation of environmental alpha radioactivity in extracted human teeth and animal bones in Malaysia

    International Nuclear Information System (INIS)

    In this study, the radiobiological analysis of natural alpha emitters in extracted human teeth and animal bones from Malaysia was estimated. The microdistributions of alpha particles in tooth and bone samples were measured using CR-39 alpha-particle track detectors. The lowest and highest alpha emission rates in teeth in the Kedah and Perak states were 0.0080 ± 0.0005 mBq cm−2 and 0.061 ± 0.008 mBq cm−2, whereas those of bones in the Perlis and Kedah states were 0.0140 ± 0.0001 mBq cm−2 and 0.7700 ± 0.0282 mBq cm−2, respectively. The average alpha emission rate in male teeth was 0.0209 ± 0.0008 mBq cm−2, whereas that of female teeth was 0.0199 ± 0.0010 mBq cm−2. The alpha emission rate in teeth is higher in smokers (0.0228 ± 0.0008 mBq cm−2) than in non-smokers (0.0179 ± 0.0008 mBq cm−2). Such difference was found statistically significant (p < 0.01). - Highlights: • Alpha emission rates in teeth from smokers slightly higher than non-smokers. • Difference between alpha rates in male and female tooth not statistically significant. • Alpha particles have the same effect at any age. • Difference between alpha rates in bones was statistically significant

  17. Utilization of wild apricot kernel press cake for extraction of protein isolate

    OpenAIRE

    Sharma, P. C.; Tilakratne, B. M. K. S.; Gupta, Anil

    2010-01-01

    The kernels of apricot (Prunus armeniaca) stones are utilized for extraction of oil. The press cake left after extraction of oil was evaluated for preparation of protein isolate for its use in food supplementation. The apricot kernels contained 45–50% oil, 23.6–26.2% protein, 4.2% ash, 5.42% crude fibre, 8.2% carbohydrates and 90 mg HCN/100 g kernels, while press cake obtained after oil extraction contained 34.5% crude protein, which can be utilized for preparation of protein isolates. The me...

  18. Physical and Biological Modification of Polycaprolactone Electrospun Nanofiber by Panax Ginseng Extract for Bone Tissue Engineering Application.

    Science.gov (United States)

    Pajoumshariati, Seyedramin; Yavari, Seyedeh Kimia; Shokrgozar, Mohammad Ali

    2016-05-01

    Medicinal plants as a therapeutic agent with osteogenic properties can enhance fracture-healing process. In this study, the osteo-inductive potential of Asian Panax Ginseng root extract within electrospun polycaprolactone (PCL) based nanofibers has been investigated. Scanning electron microscopy images revealed that all nanofibers were highly porous and beadles with average diameter ranging from 250 to 650 nm. The incorporation of ginseng extract improved the physical characteristics (i.e., hydrophilicity) of PCL nanofibers, as well as the mechanical properties. Although ginseng extract increased the degradation rate of pure PCL nanofibers, the porous structure and morphology of fibers did not change significantly after 42 days. It was found that nanofibrous scaffolds containing ginseng extract had higher proliferation (up to ~1.5 fold) compared to the pristine PCL. The qRT-PCR analysis demonstrated the addition of ginseng extract into PCL nanofibers induced significant expression of osteogenic genes (Osteocalcin, Runx-2 and Col-1) in MSCs in a concentration dependent manner. Moreover, higher calcium content, alkaline phosphatase activity and higher mineralization of MSCs were observed compared to the pristine PCL fibers. Our results indicated the promising potential of ginseng extract as an additive to enhance osteo-inductivity, mechanical and physical properties of PCL nanofibers for bone tissue engineering application. PMID:26429789

  19. Extraction of rice bran protein concentrate and its application in bread

    OpenAIRE

    W. James Harper; Sudarat Jiamyangyuen; Voranuch Srijesdaruk

    2005-01-01

    In a study of rice bran protein concentrate (RBPC) preparation, the alkaline extraction was performed. The objectives of this study were to determine the optimal extracting conditions of RBPC and apply RBPC into a bread recipe in order to improve bread quality and make acceptable product to consumers. The design used in extraction was a central composite design. The response surface methodology was chosen to graphically express the relationship between pH and extraction time with the output v...

  20. Evaluation of genetic potential of Bacopa monnieri extract in Mouse bone marrow cells by chromosomal analysis test

    Directory of Open Access Journals (Sweden)

    Shilki Vishnoi

    2013-06-01

    Full Text Available Herbs have always been used as a common source of medicines, the Bacopa monnieri is an important herb used in Aruveda as a traditional medicinal system of India. In the present investigations, the genotoxic potential of Bacopa monnieri Hydromethanolic extract (BMH was evaluated employing Chromosomal analysis assay invivo. BMH was administered to Swiss Albino mice as i.p. dose of 80mg/kg, 160mg/kg, 240mg/kg body wt., 24 hours prior the administration of cyclophosphamide (CP (positive control at the dose of 50 mg/kg body wt. A dose-dependent, significant decrease in chromosome aberration was observed with respect to control. Result suggested that BMHhave a preventive potential against CP induced chromosomal aberration in Swiss albino mouse bone marrow cells at the dose tested. Therefore seems to have a preventive potential against Chromosomal aberrations in Swiss Albinomouse bone marrow cells.

  1. Technique to assess the alveolar bone width for immediate implant placement in fresh extraction sockets

    Directory of Open Access Journals (Sweden)

    Neeraj Kumar Chandraker

    2013-01-01

    Conclusion: This technique will help the surgeon understand the thickness of labial plate especially the apical region without reflecting the flap, also aid in selection of proper dimension of dental implant, and if bone graft is needed.

  2. PPInterFinder—a mining tool for extracting causal relations on human proteins from literature

    OpenAIRE

    Raja, Kalpana; Subramani, Suresh; Natarajan, Jeyakumar

    2013-01-01

    One of the most common and challenging problem in biomedical text mining is to mine protein–protein interactions (PPIs) from MEDLINE abstracts and full-text research articles because PPIs play a major role in understanding the various biological processes and the impact of proteins in diseases. We implemented, PPInterFinder—a web-based text mining tool to extract human PPIs from biomedical literature. PPInterFinder uses relation keyword co-occurrences with protein names to extract information...

  3. Automatic extraction of gene and protein synonyms from MEDLINE and journal articles.

    OpenAIRE

    Hong YU; Hatzivassiloglou, Vasileios; Friedman, Carol; Rzhetsky, Andrey; Wilbur, W. John

    2002-01-01

    Genes and proteins are often associated with multiple names, and more names are added as new functional or structural information is discovered. Because authors often alternate between these synonyms, information retrieval and extraction benefits from identifying these synonymous names. We have developed a method to extract automatically synonymous gene and protein names from MEDLINE and journal articles. We first identified patterns authors use to list synonymous gene and protein names. We d...

  4. A simple and efficient method for PCR amplifiable DNA extraction from ancient bones

    OpenAIRE

    Kalmár, Tibor; Bachrati, Csanád Z.; Marcsik, Antónia; Raskó, István

    2000-01-01

    A simple and effective modified ethanol precipitation-based protocol is described for the preparation of DNA from ancient human bones. This method is fast and requires neither hazardous chemicals nor special devices. After the powdering and incubating of the bone samples Dextran Blue was added as a carrier for removing the PCR inhibitors with selective ethanol precipitation. This method could eliminate the time-consuming separate decalcification step, dialysis, application of centrifugation-d...

  5. Does surface anodisation of titanium implants change osseointegration and make their extraction from bone any easier?

    OpenAIRE

    Langhoff, J; Mayer, J.; Faber, L.; Kästner, S B; Guibert, G.; Zlinszky, K; Auer, J. A.; von Rechenberg, B

    2008-01-01

    Objectives: Titanium implants have a tendency for high bone-implant bonding, and, in comparison to stainless steel implants are more difficult to remove. The current study was carried out to evaluate, i) the release strength of three selected anodized titanium surfaces with increased nanohardness and low roughness, and ii) bone-implant bonding in vivo. These modified surfaces were intended to give improved anchorage while facilitating easier removal of temporary implants. Material and methods...

  6. Multifunctional Thin Film Biomatrice Biosensor in a Degradable Scaffold Containing Bone Morphogenetic Protein-2 (BMP-2) for Controlled Release in Skeletal Tissue Engineering

    Science.gov (United States)

    McDaniel, Harvey; Lomax, Linda

    2001-03-01

    Bone morphonogenetic proteins (BMP-2) have been under investigation for three decades. Deminerialized bone and extracts of deminerialized bone are o steoinductive with a temporal sequence of bone induction. Native and recombi nant BMP's have shown the ability, thru growth and differentiative factors t o induce de novo bone formation both invitro and invivo. Their principle fun ction is to induce transformation of undifferentiated mesenchymal cells into osteoblasts. Native and recombinant BMP's, when purified and used without carrier disp erse after implantation and exert no effect on bone induction. The delivery system provides the missing component to successsfully applying osteogenic p roteins for clinical need. Biological and physio-chemical properties are str ictly adhered tofor a successful delivery system. The BMP delivery system ca rrier for osteo inductive payload provided; 1)non tumorgenic genecity, 2) no n immunogenecity, 3) water insoluble, 4) biosorbability with predictable enz ymatic degradation, and 5) an optimized surface for compatibility, cell migr ation and attachment with a negative surface change that encouraged target c ell attachment. Being a controlled Release System, it binded the proteins wi th predictible BMP released kinetics. Porosity with interconnecting voids pr otected the BMP from noon specific proteolysis and promoted rapid vascular a nd mesenchymal invasion. Far wide ranging clinical applications of mechanica l and biofunctional requirements were met with the BMP delivery system. Cohe sion and malleability were reqiured forcontour augmentation, and reconstruct ion of the discontinuity defects, prevented dislocation and retained the sha pe and bone replaced the system. Biological systems have elastic activity associated with them. The activi ty was current associated with a time dependant biological/biochemical react ion (enzymic activity). Bioelectric phoenomena associated with charged molec ules in a biologic structure caused

  7. The effect of Ginkgo biloba extract treatment in the Bcl-2 expression by osteoblasts in the femoral trabecular bone of Wistar rats with glucocorticoid-induced osteoporosis

    Directory of Open Access Journals (Sweden)

    Leda M.F. Lucinda

    2014-06-01

    Full Text Available Evaluate the effect of the extract of Ginkgo biloba L., Ginkgoaceae (EGb in the Bcl-2 expression by osteoblasts in the femoral trabecular bone of Wistar rats with glucocorticoid-induced osteoporosis. Rats were divided into five groups: osteoporosis; EGb1 (28 mg/kg; EGb2 (56 mg/kg; alendronate (0.2 mg/animal and control. The treatments were conducted for 20 or 30 days. The Bcl-2 expression by osteoblasts cells was evaluated in the femoral trabecular bone. The control group was compared with the osteoporosis-induced group (Student's t-test. The other groups were analyzed by ANOVA test followed by Tukey's test (p < 0.05. The percentage of Bcl-2 expression was reduced, when the control group (17.95 ± 3.45 20 days; 21.11 ± 3.43 30 days was compared with the osteoporosis group (10.64 ± 3.30 20 days; 9.72 ± 2.84 30 days. Nevertheless, this percentage increased in the EGb2 group (18.58 ± 3.41 20 days; 16.51 ± 1.80 30 days when compared to the osteoporosis group. The EGb increased the expression of the anti-apoptotic protein, suggesting a decrease in osteoblast apoptosis.

  8. DSP-PP precursor protein cleavage by tolloid-related-1 protein and by bone morphogenetic protein-1.

    Directory of Open Access Journals (Sweden)

    Helena H Ritchie

    Full Text Available Dentin sialoprotein (DSP and phosphophoryn (PP, acidic proteins critical to dentin mineralization, are translated from a single transcript as a DSP-PP precursor that undergoes specific proteolytic processing to generate DSP and PP. The cleavage mechanism continues to be controversial, in part because of the difficulty of obtaining DSP-PP from mammalian cells and dentin matrix. We have infected Sf9 cells with a recombinant baculovirus to produce large amounts of secreted DSP-PP(240, a variant form of rat DSP-PP. Mass spectrometric analysis shows that DSP-PP(240 secreted by Sf9 cells undergoes specific cleavage at the site predicted from the N-terminal sequence of PP extracted from dentin matrix: SMQG(447↓D(448DPN. DSP-PP(240 is cleaved after secretion by a zinc-dependent activity secreted by Sf9 cells, generating DSP(430 and PP(240 products that are stable in the medium. DSP-PP processing activity is constitutively secreted by Sf9 cells, but secretion is diminished 3 days after infection. Using primers corresponding to the highly conserved catalytic domain of Drosophila melanogaster tolloid (a mammalian BMP1 homolog, we isolated a partial cDNA for a Spodopotera frugiperda tolloid-related-1 protein (TLR1 that is 78% identical to Drosophila TLR1 but only 65% identical to Drosophila tolloid. Tlr1 mRNA decreased rapidly in Sf9 cells after baculovirus infection and was undetectable 4d after infection, paralleling the observed decrease in secretion of the DSP-PP(240 processing activity after infection. Human BMP1 is more similar to Sf9 and Drosophila TLR1 than to tolloid, and Sf9 TLR1 is more similar to BMP1 than to other mammalian homologs. Recombinant human BMP1 correctly processed baculovirus-expressed DSP-PP(240 in a dose-dependent manner. Together, these data suggest that the physiologically accurate cleavage of mammalian DSP-PP(240 in the Sf9 cell system represents the action of a conserved processing enzyme and support the proposed role of BMP

  9. Pressurized water extraction of isoflavones by experimental design from soybean flour and Soybean Protein Isolate.

    Science.gov (United States)

    Moras, Benjamin; Rey, Stéphane; Vilarem, Gérard; Pontalier, Pierre-Yves

    2017-01-01

    A Doehlert experimental design was conducted and surface response methodology was used to determine the effect of temperature, contact time and solid liquid ratio on isoflavone extraction from soybean flour or Soybean Protein Isolate in pressurized water system. The optimal conditions conducted gave an extraction yield of 85% from soybean flour. For Soybean Protein Isolate compared to soybean flour, the isoflavone extraction yield is 61%. This difference could be explained by higher aglycon content, while aglycon appears to be the least extracted isoflavone by pressurized water. The solid liquid ratio in the ASE cell was the overriding factor in obtaining high yields with both soybean products, while temperature has less influence. A high temperature causes conversion of the malonyls-glucosides and glucosides isoflavone derivatives into glucosides or aglycons forms. pressurized water extraction showed a high solubilization of protein material up to 95% of inserted Soybean Protein Isolate. PMID:27507441

  10. Anticariogenic and Hemolytic Activity of Selected Seed Protein Extracts In vitro conditions.

    Directory of Open Access Journals (Sweden)

    Kalpesh B Ishnava

    2014-10-01

    Full Text Available This study aimed to assess the anticariogenic and hemolytic activity of crude plant seed protein extracts against tooth decaying bacteria.The proteins from seeds of 12 different plants were extracted and used for antimicrobial assay against six different organisms. The extraction was carried out in 10mM of sodium phosphate buffer (pH 7.0. Protein concentrations were determined as described by Bradford method. Anticariogenic activity was studied by agar well diffusion method and Minimum Inhibitory Concentration (MIC was evaluated by the two-fold serial broth dilution method. Hemolytic activity, treatment of proteinase K and Kinetic study in Mimusops elengi crude seed protein extract.The anticariogenic assay demonstrated the activity of Mimusops elengi against Staphylococcus aureus and Streptococcus pyogenes. A minor activity of Glycine wightii against Streptococcus mutans was also found. The protein content of Mimusops elengi seed protein extract was 5.84mg/ml. The MIC values for Staphylococcus aureus and Streptococcus pyogenes against Mimusops elengi seed protein extract were 364.36μg/ml and 182.19μg/ml, respectively. Kinetic study further elucidated the mode of inhibition in the presence of the Mimusops elengi plant seed protein with respect to time. The concentration of crude extract which gave 50% hemolysis compared to Triton X-100 treatment (HC50 value was 1.58 mg/ml; which is more than five times larger than that of the MIC. Treatment with proteinase K of the Mimusops elengi seed protein resulted in absence of the inhibition zone; which clearly indicates that the activity was only due to protein.Our results showed the prominence of Mimusops elengi plant seed protein extract as an effective herbal medication against tooth decaying bacteria.

  11. Effect of 5-azacytidine on the Protein Expression of Porcine Bone Marrow Mesenchymal Stem Cells in vitro

    Institute of Scientific and Technical Information of China (English)

    Neng-Sheng Ye; Rong-Li Zhang; Yan-Feng Zhao; Xue Feng; Yi-Ming Wang; Guo-An Luo

    2006-01-01

    Bone marrow-derived mesenchymal stem cells (MSCs) are pluripotent stem cells that show a vital potential in the clinical application for cell transplantation. In the present paper, proteomic techniques were used to approach the protein profiles associated with porcine bone marrow MSCs and investigate the regulation of MSC proteins on the effect of 5-azacytidine (5-aza). Over 1,700 protein species were separated from MSCs according to gel analysis. Compared with the expression profiling of control MSCs, there were 11 protein spots up-regulated and 26 downregulated in the protein pattern of 5-aza-treated cells. A total of 21 proteins were successfully identified by MALDI-TOF-MS analysis, among which some interesting proteins, such as alpha B-crystallin, annexin A2, and stathmin 1, had been reported to involve in cell proliferation and differentiation through different signaling pathways. Our data should be useful for the future study of MSC differentiation and apoptosis.

  12. Parathyroid hormone-related protein (PTHrP) expression and bone invasion by oral squamous cell carcinoma

    International Nuclear Information System (INIS)

    Parathyroid hormone-related protein (PTHrP) indirectly stimulates osteoclastic bone resorption through osteoblasts in humoral hypercalcemia of malignancy. We reported that the serum concentration of PTHrP elevated in terminal stage patients with oral squamous cell carcinoma (SCC) in 1996. Therefore, PTHrP is a candidate for direct bone resorption factor released from the tumor tissue. The purpose of this study was to elucidate the correlation between the direct bone invasion by oral SCC and PTHrP expression. The serum C-PTHrP concentration was measured in 53 patients with oral SCC. The immunohistochemical study using PTHrP (labeled streptoavidin-biotin method, 38-64 monoclonal and 1-34 polyclonal antibody) was performed in 53 biopsy specimens. The bone invasion was assessed by using panoramic radiographs and bone scintigrams (99mTc-MDP). The mean serum C-PTHrP concentration in the bone invasion identified group was 43.1±17.2 pmol/1. In the non-bone invasion group it was 42.0±18.0 pmol/1. No significant correlation was found between serum C-PTHrP levels and bone invasion or between PTHrP (1-34) and (38-64) expression in tumors and bone invasion. These results showed that there is no relationship between PTHrP expression in the biopsy specimen and direct bone invasion. Since the expression of PTHrP in the tumor tissue attached to the bone or surgical specimens has not been investigated, it is still unclear if PTHrP plays a role in direct bone resorption by oral SCC. (author)

  13. Bone morphogenetic protein in complex cervical spine surgery: A safe biologic adjunct?

    OpenAIRE

    Lebl, Darren R.

    2013-01-01

    The advent of recombinant DNA technology has substantially increased the intra-operative utilization of biologic augmentation in spine surgery over the past several years after the Food and Drug Administration approval of the bone morphogenetic protein (BMP) class of molecules for indications in the lumbar spine. Much less is known about the potential benefits and risks of the “off-label” use of BMP in the cervical spine. The history and relevant literature pertaining to the use of the “off-l...

  14. Effect of compounds astragalus extract and cysteamine on finishing pigs growth and protein turnover

    International Nuclear Information System (INIS)

    32 PIC pigs, weighted 59.82±1.32 kg on average, were subjected randomly to 4 treatments for determining the effects of extrogenous metabolism-regulating agents on pig performance, protein turn-over and nitrogen retention by using isotope (15N-Gly) technique and nitrogen balance trial. The 4 treatments were: (1) basal diet; (2) basal diet plus cysteamine (70mg/kg, CS group); (3) basal diet plus non-stable astragalus extracts 250mg/kg (NCAE group); (4) basal diet plus stably astragalus extracts 250mg/kg (CAE group). Astragalus extracts and cysteamine, especially NCAE, redounded to increase pig performance, especially increase average day gain (P<0.05). Both astragalus extracts and cysteamine accelerated protein aggradation rate and amino acid utilization rate, as well as biological value. Astragalus extracts helped to accelerate the synthesis rate of amino acid into body protein and reduce the excretion rate of endogenous urine nitrogen. The mechanism of increasing protein deposition of cysteamine is reducing body protein degradation rate (26.71% reduction, compared to control group), while that of astragalus extracts is bilateral, reducing protein degradation rate (24.84% and 3.66% reduction for NCAE and CAE) and accelerating protein synthesis rate (22.86% and 19.18% increase for NCAE and CAE). Both astragalus extracts and cysteamine contributed the increasing nitrogen retention, net nitrogen utilization and biological value. (authors)

  15. Soy protein is beneficial but high-fat diet and voluntary running are detrimental to bone structure in mice

    Science.gov (United States)

    We investigated the effects of diet (AIN93G or high-fat), physical activity (sedentary or voluntary running) and protein source (casein or soy protein isolate) and their interactions on bone microstructural changes in distal femurs in male C57BL/6 mice by using micro-computed tomography. After 14 w...

  16. Improved Proteomic Analysis Following Trichloroacetic Acid Extraction of Bacillus anthracis Spore Proteins

    Energy Technology Data Exchange (ETDEWEB)

    Kaiser, Brooke LD; Wunschel, David S.; Sydor, Michael A.; Warner, Marvin G.; Wahl, Karen L.; Hutchison, Janine R.

    2015-08-07

    Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Proteomic analysis is dependent upon efficient extraction of proteins from bacterial samples without introducing bias toward extraction of particular protein classes. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrich for certain classes of proteins. The method presented here is technically simple and does not require specialized equipment such as a mechanical disrupter. Our data reveal that for particularly challenging samples, such as B. anthracis Sterne spores, trichloroacetic acid extraction improved the number of proteins identified within a sample compared to bead beating (714 vs 660, respectively). Further, TCA extraction enriched for 103 known spore specific proteins whereas bead beating resulted in 49 unique proteins. Analysis of C. botulinum samples grown to 5 days, composed of vegetative biomass and spores, showed a similar trend with improved protein yields and identification using our method compared to bead beating. Interestingly, easily lysed samples, such as B. anthracis vegetative cells, were equally as effectively processed via TCA and bead beating, but TCA extraction remains the easiest and most cost effective option. As with all assays, supplemental methods such as implementation of an alternative preparation method may provide additional insight to the protein biology of the bacteria being studied.

  17. Abnormal IGF-Binding Protein Profile in the Bone Marrow of Multiple Myeloma Patients.

    Directory of Open Access Journals (Sweden)

    Liesbeth Bieghs

    Full Text Available Insulin-like growth factor (IGF signalling plays a key role in homing, progression, and treatment resistance in multiple myeloma (MM. In the extracellular environment, the majority of IGF molecules are bound to one of six IGF-binding proteins (IGFBP1-6, leaving a minor fraction of total IGF free and accessible for receptor activation. In MM, high IGF-receptor type 1 expression levels correlate with a poor prognosis, but the status and role of IGF and IGFBPs in the pathobiology of MM is unknown. Here we measured total IGF1, IGF2, and intact IGFBP levels in blood and bone marrow samples from MM (n = 17, monoclonal gammopathy of undetermined significance (MGUS (n = 37, and control individuals (n = 15, using ELISA (IGFs and 125I-IGF1 Western Ligand Blotting (IGFBPs. MGUS and MM patients displayed a significant increase in intact IGFBP-2 (2.5-3.8 fold and decrease in intact IGFBP-3 (0.6-0.5 fold in the circulation compared to control individuals. Further, IGFBP-2 as well as total IGFBP levels were significantly lower in bone marrow compared to circulation in MM and MGUS only, whereas IGF1, IGF2, and IGFBP-3 were equally distributed between the two compartments. In conclusion, the profound change in IGFBP profile strongly suggests an increased IGF bioavailability in the bone marrow microenvironment in MGUS and MM, despite no change in growth factor concentration.

  18. Abnormal IGF-Binding Protein Profile in the Bone Marrow of Multiple Myeloma Patients.

    Science.gov (United States)

    Bieghs, Liesbeth; Brohus, Malene; Kristensen, Ida B; Abildgaard, Niels; Bøgsted, Martin; Johnsen, Hans E; Conover, Cheryl A; De Bruyne, Elke; Vanderkerken, Karin; Overgaard, Michael T; Nyegaard, Mette

    2016-01-01

    Insulin-like growth factor (IGF) signalling plays a key role in homing, progression, and treatment resistance in multiple myeloma (MM). In the extracellular environment, the majority of IGF molecules are bound to one of six IGF-binding proteins (IGFBP1-6), leaving a minor fraction of total IGF free and accessible for receptor activation. In MM, high IGF-receptor type 1 expression levels correlate with a poor prognosis, but the status and role of IGF and IGFBPs in the pathobiology of MM is unknown. Here we measured total IGF1, IGF2, and intact IGFBP levels in blood and bone marrow samples from MM (n = 17), monoclonal gammopathy of undetermined significance (MGUS) (n = 37), and control individuals (n = 15), using ELISA (IGFs) and 125I-IGF1 Western Ligand Blotting (IGFBPs). MGUS and MM patients displayed a significant increase in intact IGFBP-2 (2.5-3.8 fold) and decrease in intact IGFBP-3 (0.6-0.5 fold) in the circulation compared to control individuals. Further, IGFBP-2 as well as total IGFBP levels were significantly lower in bone marrow compared to circulation in MM and MGUS only, whereas IGF1, IGF2, and IGFBP-3 were equally distributed between the two compartments. In conclusion, the profound change in IGFBP profile strongly suggests an increased IGF bioavailability in the bone marrow microenvironment in MGUS and MM, despite no change in growth factor concentration. PMID:27111220

  19. cAMP-response-element-binding protein positively regulates breast cancer metastasis and subsequent bone destruction

    Energy Technology Data Exchange (ETDEWEB)

    Son, Jieun; Lee, Jong-Ho; Kim, Ha-Neui; Ha, Hyunil, E-mail: hyunil74@hotmail.com; Lee, Zang Hee, E-mail: zang1959@snu.ac.kr

    2010-07-23

    Research highlights: {yields} CREB is highly expressed in advanced breast cancer cells. {yields} Tumor-related factors such as TGF-{beta} further elevate CREB expression. {yields} CREB upregulation stimulates metastatic potential of breast cancer cells. {yields} CREB signaling is required for breast cancer-induced bone destruction. -- Abstract: cAMP-response-element-binding protein (CREB) signaling has been reported to be associated with cancer development and poor clinical outcome in various types of cancer. However, it remains to be elucidated whether CREB is involved in breast cancer development and osteotropism. Here, we found that metastatic MDA-MB-231 breast cancer cells exhibited higher CREB expression than did non-metastatic MCF-7 cells and that CREB expression was further increased by several soluble factors linked to cancer progression, such as IL-1, IGF-1, and TGF-{beta}. Using wild-type CREB and a dominant-negative form (K-CREB), we found that CREB signaling positively regulated the proliferation, migration, and invasion of MDA-MB-231 cells. In addition, K-CREB prevented MDA-MB-231 cell-induced osteolytic lesions in a mouse model of cancer metastasis. Furthermore, CREB signaling in cancer cells regulated the gene expression of PTHrP, MMPs, and OPG, which are closely involved in cancer metastasis and bone destruction. These results indicate that breast cancer cells acquire CREB overexpression during their development and that this CREB upregulation plays an important role in multiple steps of breast cancer bone metastasis.

  20. Immune targeting of fibroblast activation protein triggers recognition of multipotent bone marrow stromal cells and cachexia

    Science.gov (United States)

    Chinnasamy, Dhanalakshmi; Yu, Zhiya; Morgan, Richard A.; Lee, Chyi-Chia Richard; Restifo, Nicholas P.

    2013-01-01

    Fibroblast activation protein (FAP) is a candidate universal target antigen because it has been reported to be selectively expressed in nearly all solid tumors by a subset of immunosuppressive tumor stromal fibroblasts. We verified that 18/18 human tumors of various histologies contained pronounced stromal elements staining strongly for FAP, and hypothesized that targeting tumor stroma with FAP-reactive T cells would inhibit tumor growth in cancer-bearing hosts. T cells genetically engineered with FAP-reactive chimeric antigen receptors (CARs) specifically degranulated and produced effector cytokines upon stimulation with FAP or FAP-expressing cell lines. However, adoptive transfer of FAP-reactive T cells into mice bearing a variety of subcutaneous tumors mediated limited antitumor effects and induced significant cachexia and lethal bone toxicities in two mouse strains. We found that FAP was robustly expressed on PDGFR-α+, Sca-1+ multipotent bone marrow stromal cells (BMSCs) in mice, as well as on well-characterized, clinical-grade multipotent human BMSCs. Accordingly, both mouse and human multipotent BMSCs were recognized by FAP-reactive T cells. The lethal bone toxicity and cachexia observed after cell-based immunotherapy targeting FAP cautions against its use as a universal target. Moreover, the expression of FAP by multipotent BMSCs may point toward the cellular origins of tumor stromal fibroblasts. PMID:23712432

  1. Extractable proteins from electron beam (EB) irradiated natural rubber (NR) latex

    International Nuclear Information System (INIS)

    The protein assay of natural rubber latex (NRL) vulcanized by low energy electron beam (EB, 300 keV, 30 mA) has been carried out using Bicinchoninic acid (BCA) reagent. Extractable protein in irradiated latex film was determined by measuring the absorption of colored solution at 562 nm using UV spectrometer. The effect of various radiation doses on the extractable protein content of NRL was investigated. It was ,found that the quantities of extractable protein increases with radiation dose. When compared with ,gamma-ray irradiated samples the same trend was observed. Electron beam irradiated latex films are leached in 1% (ammonia water for various lengths of time. From the results it was established that within 2 hours of leaching in ammonia water most of the extractable protein (96%) were removed from rubber film

  2. PROTEIN EXTRACTION OF MAIZE (ZEA MAYS FOR PROTEOMIC 2 – DE ANALYSIS

    Directory of Open Access Journals (Sweden)

    Edita Gregová

    2014-12-01

    Full Text Available A strength of two-dimensional polyacrylamide gel electrophoresis (2D PAGE is its ability to resolve and investigate the abundance of several thousand proteins in a single sample. Two different extraction procedures for two-dimensional electrophoresis of plant proteins are compared in this work. Phenol-based extraction methods have been mainly used to extract proteins from different organs or tissues on many species. We wanted to determine which of these protocols was optimal for starch plants in order to achieve both efficient protein extraction and high spot resolution on 2-D gels. The phenol-based protocol was superior to the sodium phosphatase methods, showing larger protein yields and greater spot resolution on 2-D gels.

  3. Extraction and clipping repair of a chicken bone penetrating the gastric wall

    Institute of Scientific and Technical Information of China (English)

    Jin-Soo Kim; Hyung-Keun Kim; Young-Seok Cho; Hiun-Suk Chae; Chang-Whan Kim; Byung-Wook Kim; Sok-Won Han; Kyu-Yong Choi

    2008-01-01

    We report a case of gastric penetration caused by accidental ingestion of a chicken bone in a 42-year old woman with a partially wearing denture. Three days ago, she accidentally swallowed several lumps of poorly-chewed chicken. Physical examination disclosed mild tenderness in the periumbilical area. Abdominal Computed tomography (CT) showed a suspicious penetration or perforation of the stomach wall measuring about 3 cm, by a linear radiopaque material at the lesser curvature of the antrum. The end of a chicken bone was very close to but did not penetrate the liver. Endoscopic examination revealed a chicken bone that penetrated into the prepyloric antrum. The penetrating chicken bone was removed with grasping forceps. Five endoscopic clips were applied immediately at the removal site and the periumbilical pain resolved promptly. After removal of the chicken bone, the patient was treated with conservative care for three days, after which she was completely asymptomatic and discharged without complication. To treat gastric penetration by a foreign body, endoclipping can be a useful method in patients with no signs or symptoms of peritoneal irritation.

  4. Osteogenic efficiency of in situ gelling poloxamine systems with and without bone morphogenetic protein-2

    Directory of Open Access Journals (Sweden)

    A Rey-Rico

    2011-04-01

    Full Text Available In situ gelling solutions for minimally invasive local application of bone growth factors are attracting increasing attention as efficient and patient-friendly alternative to bone grafts and solid scaffolds for repairing bone defects. Poloxamines, i.e., X-shaped poly(ethylene oxide-poly(propylene oxide block copolymers with an ethylenediamine core (Tetronic®, were evaluated both as an active osteogenic component and as a vehicle for rhBMP-2 injectable implants. After cytotoxicity screening of various poloxamine varieties, Tetronic 908, 1107, 1301 and 1307 solutions were chosen as the most cytocompatible and their sol-to-gel transitions were rheologically characterized. Viscoelastic gels, formed at 37 ºC, sustained protein release under physiological-like conditions. Formulations of rhBMP-2 led to differentiation of mesenchymal stem cells to osteoblasts, quantified as alkaline phosphatase activity with a maximum at day 7, and to mineralized nodules. Interestingly, poloxamine solely gels led to an initial proliferation of the mesenchymal stem cells (first week, followed by differentiation to osteoblasts (second to third week. Histochemical analysis revealed that Tetronic 908 is only osteoinductive; Tetronic 1107 is mostly osteoinductive, although its use leads to a minor differentiation to adipocytes; Tetronic 1307, solely or loaded with rhBMP-2, causes differentiation of both osteoblasts and adipocytes. Enhanced expression levels of CBFA-1 and collagen type I were observed for Tetronic 908, 1107 and 1307, both solely and combined with rhBMP-2. The intrinsic osteogenic activity of poloxamines (not observed for Pluronic F127 offers novel perspectives for bone regeneration using minimally invasive procedures (i.e., injectable scaffolds and overcoming the safety and the cost/effectiveness concerns associated with large scale clinical use of recombinant growth factors.

  5. The Use of Bone Morphogenetic Protein in Pediatric Cervical Spine Fusion Surgery: Case Reports and Review of the Literature

    OpenAIRE

    Molinari, Robert W.; Molinari, Christine

    2015-01-01

    Study Design Case report. Objective There is a paucity of literature describing the use of bone graft substitutes to achieve fusion in the pediatric cervical spine. The outcomes and complications involving the off-label use of bone morphogenetic protein (BMP)-2 in the pediatric cervical spine are not clearly defined. The purpose of this article is to report successful fusion without complications in two pediatric patients who had instrumented occipitocervical fusion using low-dose BMP-2. Meth...

  6. Altered protein secretions during interactions between adipose tissue- or bone marrow-derived stromal cells and inflammatory cells

    OpenAIRE

    Hattori, Hidemi; Ishihara, Masayuki

    2015-01-01

    Introduction Paracrine effects can be exploited in cell-based therapies that secrete factors, such as chemokines and cytokines, and can recruit inflammatory cells to transplants. In this study, mouse adipose tissue-derived stromal cells (ASCs) and bone marrow-derived stromal cells (ST2 cells) were used to examine changes in paracrine interactions with inflammation cells. Methods Green fluorescent protein positive (GFP+) bone marrow cells (BMCs) were injected into an irradiated mouse via the f...

  7. Bone morphogenetic proteins regulate osteoprotegerin and its ligands in human vascular smooth muscle cells

    DEFF Research Database (Denmark)

    Knudsen, Kirsten Quyen Nguyen; Olesen, Ping; Ledet, Thomas;

    2007-01-01

    ) and TNF-related apoptosis-inducing ligand (TRAIL) in HVSMC. All three growth factors decreased OPG protein production significantly; these results were paralleled by reduced OPG mRNA expression. TRAIL mRNA levels were also decreased. RANKL mRNA expression declined when treated with TGF-beta1 but were......The bone-related protein osteoprotegerin (OPG) may be involved in the development of vascular calcifications, especially in diabetes, where it has been found in increased amounts in the arterial wall. Experimental studies suggest that members of the TGF-superfamily are involved in the...... transformation of human vascular smooth muscle cells (HVSMC) to osteoblast-like cells. In this study, we evaluated the effect of BMP-2, BMP-7 and transforming growth factor beta (TGF-beta1) on the secretion and mRNA expression of OPG and its ligands receptor activator of nuclear factor-kappabeta ligand (RANKL...

  8. A green deep eutectic solvent-based aqueous two-phase system for protein extracting

    International Nuclear Information System (INIS)

    Highlights: • A strategy for the protein purification with a deep eutectic solvent(DES)-based aqueous two-phase system. • Choline chloride-glycerin DES was selected as the extraction solvent. • Bovine serum albumin and trypsin were used as the analytes. • Aggregation phenomenon was detected in the mechanism research. - Abstract: As a new type of green solvent, deep eutectic solvent (DES) has been applied for the extraction of proteins with an aqueous two-phase system (ATPS) in this work. Four kinds of choline chloride (ChCl)-based DESs were synthesized to extract bovine serum albumin (BSA), and ChCl-glycerol was selected as the suitable extraction solvent. Single factor experiments have been done to investigate the effects of the extraction process, including the amount of DES, the concentration of salt, the mass of protein, the shaking time, the temperature and PH value. Experimental results show 98.16% of the BSA could be extracted into the DES-rich phase in a single-step extraction under the optimized conditions. A high extraction efficiency of 94.36% was achieved, while the conditions were applied to the extraction of trypsin (Try). Precision, repeatability and stability experiments were studied and the relative standard deviations (RSD) of the extraction efficiency were 0.4246% (n = 3), 1.6057% (n = 3) and 1.6132% (n = 3), respectively. Conformation of BSA was not changed during the extraction process according to the investigation of UV–vis spectra, FT-IR spectra and CD spectra of BSA. The conductivity, dynamic light scattering (DLS) and transmission electron microscopy (TEM) were used to explore the mechanism of the extraction. It turned out that the formation of DES–protein aggregates play a significant role in the separation process. All the results suggest that ChCl-based DES-ATPS are supposed to have the potential to provide new possibilities in the separation of proteins

  9. A green deep eutectic solvent-based aqueous two-phase system for protein extracting

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Kaijia; Wang, Yuzhi, E-mail: wyzss@hnu.edu.cn; Huang, Yanhua; Li, Na; Wen, Qian

    2015-03-15

    Highlights: • A strategy for the protein purification with a deep eutectic solvent(DES)-based aqueous two-phase system. • Choline chloride-glycerin DES was selected as the extraction solvent. • Bovine serum albumin and trypsin were used as the analytes. • Aggregation phenomenon was detected in the mechanism research. - Abstract: As a new type of green solvent, deep eutectic solvent (DES) has been applied for the extraction of proteins with an aqueous two-phase system (ATPS) in this work. Four kinds of choline chloride (ChCl)-based DESs were synthesized to extract bovine serum albumin (BSA), and ChCl-glycerol was selected as the suitable extraction solvent. Single factor experiments have been done to investigate the effects of the extraction process, including the amount of DES, the concentration of salt, the mass of protein, the shaking time, the temperature and PH value. Experimental results show 98.16% of the BSA could be extracted into the DES-rich phase in a single-step extraction under the optimized conditions. A high extraction efficiency of 94.36% was achieved, while the conditions were applied to the extraction of trypsin (Try). Precision, repeatability and stability experiments were studied and the relative standard deviations (RSD) of the extraction efficiency were 0.4246% (n = 3), 1.6057% (n = 3) and 1.6132% (n = 3), respectively. Conformation of BSA was not changed during the extraction process according to the investigation of UV–vis spectra, FT-IR spectra and CD spectra of BSA. The conductivity, dynamic light scattering (DLS) and transmission electron microscopy (TEM) were used to explore the mechanism of the extraction. It turned out that the formation of DES–protein aggregates play a significant role in the separation process. All the results suggest that ChCl-based DES-ATPS are supposed to have the potential to provide new possibilities in the separation of proteins.

  10. Effects of a Mikania laevigata extract on bone resorption and RANKL expression during experimental periodontitis in rats

    Directory of Open Access Journals (Sweden)

    Bruno B. Benatti

    2012-06-01

    Full Text Available OBJECTIVES: The Mikania laevigata extract (MLE (popularly known in Brazil as "guaco" possesses anti-inflammatory properties. In the present study we tested the effects of MLE in a periodontitis experimental model in rats. We also investigated possible mechanisms underlying such effects. MATERIAL AND METHODS: Periodontal disease was induced by a ligature placed around the mandibular first molars of each animal. Male Wistar rats were divided into 4 groups: non-ligated animals treated with vehicle; non-ligated animals treated with MLE (10 mg/kg, daily; ligature-induced animals treated with vehicle and ligature-induced animals treated with MLE (10 mg/kg, daily. Thirty days after the induction of periodontal disease, the animals were euthanized and mandibles and gingival tissues removed for further analysis. RESULTS: Morphometric analysis of alveolar bone loss demonstrated that MLE-treated animals presented a decreased alveolar bone loss and a lower expression of the activator of nuclear factor-κB ligand (RANKL measured by immunohistochemistry. Moreover, gingival tissues from the MLE-treated group showed decreased neutrophil migration myeloperoxidase (MPO assay. CONCLUSIONS: These results indicate that MLE may be useful to control bone resorption during progression of experimental periodontitis in rats.

  11. In Vivo Chemoprotective Activity of Bovine Dialyzable Leukocyte Extract in Mouse Bone Marrow Cells against Damage Induced by 5-Fluorouracil

    Science.gov (United States)

    Coronado-Cerda, Erika Evangelina; Franco-Molina, Moisés Armides; Mendoza-Gamboa, Edgar; Prado-García, Heriberto; Rivera-Morales, Lydia Guadalupe; Zapata-Benavides, Pablo; Rodríguez-Salazar, María del Carmen; Caballero-Hernandez, Diana; Tamez-Guerra, Reyes Silvestre; Rodríguez-Padilla, Cristina

    2016-01-01

    Chemotherapy treatments induce a number of side effects, such as leukopenia neutropenia, peripheral erythropenia, and thrombocytopenia, affecting the quality of life for cancer patients. 5-Fluorouracil (5-FU) is wieldy used as myeloablative model in mice. The bovine dialyzable leukocyte extract (bDLE) or IMMUNEPOTENT CRP® (ICRP) is an immunomodulatory compound that has antioxidants and anti-inflammatory effects. In order to investigate the chemoprotection effect of ICRP on bone marrow cells in 5-FU treated mice, total bone marrow (BM) cell count, bone marrow colony forming units-granulocyte/macrophage (CFU-GM), cell cycle, immunophenotypification, ROS/superoxide and Nrf2 by flow cytometry, and histological and hematological analyses were performed. Our results demonstrated that ICRP increased BM cell count and CFU-GM number, arrested BM cells in G0/G1 phase, increased the percentage of leukocyte, granulocytic, and erythroid populations, reduced ROS/superoxide formation and Nrf2 activation, and also improved hematological levels and weight gain in 5-FU treated mice. These results suggest that ICRP has a chemoprotective effect against 5-FU in BM cells that can be used in cancer patients. PMID:27191003

  12. Comparison of Two Methods for the Extraction of Fractionated Rice Bran Protein

    Directory of Open Access Journals (Sweden)

    Changyuan Wang

    2014-01-01

    Full Text Available Two different methods for extracting fractionated rice bran protein (FRBP from defatted rice bran were investigated according to the solubility of protein in different extraction solvents. The yields of the obtained proteins and their purity were first compared. Sodium dodecyl sulfate polyacrylamide gel electrophoresis, differential scanning calorimetry, protein surface hydrophobicity, and protein secondary molecular structure analyses were subsequently applied to identify and compare the compositional, structural, and functional characteristics of the obtained proteins. The highest yield (13.8%, w/w and purity (45–47% of FRBP products were obtained using 0.4 M NaCl, 80% ethanol, and 0.01 M NaOH as extraction solvents to fractionate albumin, globulin, prolamin, and glutelin. Several good properties were exhibited, including good functionality, specific denaturation temperature, and enthalpy values, for FRBP products prepared by the above method.

  13. On-Orbit DNA, RNA, and Protein Extraction Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Genova Engineering proposes to develop and demonstrate a toolset of discrete devices and extraction kits which will leverage existing on-orbit facilities and will...

  14. Makings of a brittle bone: Unexpected lessons from a low protein diet study of a mouse OI model.

    Science.gov (United States)

    Mertz, E L; Makareeva, E; Mirigian, L S; Koon, K Y; Perosky, J E; Kozloff, K M; Leikin, S

    2016-01-01

    Glycine substitutions in type I collagen appear to cause osteogenesis imperfecta (OI) by disrupting folding of the triple helix, the structure of which requires Gly in every third position. It is less clear, however, whether the resulting bone malformations and fragility are caused by effects of intracellular accumulation of misfolded collagen on differentiation and function of osteoblasts, effects of secreted misfolded collagen on the function of bone matrix, or both. Here we describe a study originally conceived for testing how reducing intracellular accumulation of misfolded collagen would affect mice with a Gly610 to Cys substitution in the triple helical region of the α2(I) chain. To stimulate degradation of misfolded collagen by autophagy, we utilized a low protein diet. The diet had beneficial effects on osteoblast differentiation and bone matrix mineralization, but also affected bone modeling and suppressed overall animal growth. Our more important observations, however, were not related to the diet. They revealed how altered osteoblast function and deficient bone formation by each cell caused by the G610C mutation combined with increased osteoblastogenesis might make the bone more brittle, all of which are common OI features. In G610C mice, increased bone formation surface compensated for reduced mineral apposition rate, resulting in normal cortical area and thickness at the cost of altering cortical modeling process, retaining woven bone, and reducing the ability of bone to absorb energy through plastic deformation. Reduced collagen and increased mineral density in extracellular matrix of lamellar bone compounded the problem, further reducing bone toughness. The latter observations might have particularly important implications for understanding OI pathophysiology and designing more effective therapeutic interventions. PMID:27039252

  15. Cross talk between insulin and bone morphogenetic protein signaling systems in brown adipogenesis

    DEFF Research Database (Denmark)

    Zhang, Hongbin; Schulz, Tim J; Espinoza, Daniel O;

    2010-01-01

    Both insulin and bone morphogenetic protein (BMP) signaling systems are important for adipocyte differentiation. Analysis of gene expression in BMP7-treated fibroblasts revealed a coordinated change in insulin signaling components by BMP7. To further investigate the cross talk between insulin and...... BMP7's suppressive effect on pref-1 transcription. Together, these data suggest cross talk between the insulin and BMP signaling systems by which BMP7 can rescue brown adipogenesis in cells with insulin resistance.......Both insulin and bone morphogenetic protein (BMP) signaling systems are important for adipocyte differentiation. Analysis of gene expression in BMP7-treated fibroblasts revealed a coordinated change in insulin signaling components by BMP7. To further investigate the cross talk between insulin and...... BMP signaling systems in brown adipogenesis, we examined the effect of BMP7 in insulin receptor substrate 1 (IRS-1)-deficient brown preadipocytes, which exhibit a severe defect in differentiation. Treatment of these cells with BMP7 for 3 days prior to adipogenic induction restored differentiation and...

  16. 3-Dimensional cell-laden nano-hydroxyapatite/protein hydrogels for bone regeneration applications

    Energy Technology Data Exchange (ETDEWEB)

    Sadat-Shojai, Mehdi, E-mail: msadatshojai@gmail.com [Department of Chemistry, College of Sciences, Shiraz University, Shiraz 71454 (Iran, Islamic Republic of); Department of Biomaterials, Iran Polymer and Petrochemical Institute, Tehran (Iran, Islamic Republic of); Khorasani, Mohammad-Taghi [Department of Biomaterials, Iran Polymer and Petrochemical Institute, Tehran (Iran, Islamic Republic of); Jamshidi, Ahmad [Department of Novel Drug Delivery Systems, Iran Polymer and Petrochemical Institute, Tehran (Iran, Islamic Republic of)

    2015-04-01

    The ability to encapsulate cells in three-dimensional (3D) protein-based hydrogels is potentially of benefit for tissue engineering and regenerative medicine. However, as a result of their poor mechanical strength, protein-based hydrogels have traditionally been considered for soft tissue engineering only. Hence, in this study we tried to render these hydrogels suitable for hard tissue regeneration, simply by incorporation of bioactive nano-hydroxyapatite (HAp) into a photocrosslinkable gelatin hydrogel. Different cell types were also encapsulated in three dimensions in the resulting composites to prepare cell-laden constructs. According to the results, HAp significantly improves the stiffness of gelatin hydrogels, while it maintains their structural integrity and swelling ratio. It was also found that while the bare hydrogel (control) was completely inert in terms of bioactivity, a homogeneous 3D mineralization occurs throughout the nanocomposites after incubation in simulated body fluid. Moreover, encapsulated cells readily elongated, proliferated, and formed a 3D interconnected network with neighboring cells in the nanocomposite, showing the suitability of the nano-HAp/protein hydrogels for cellular growth in 3D. Therefore, the hydrogel nanocomposites developed in this study may be promising candidates for preparing cell-laden tissue-like structures with enhanced stiffness and increased osteoconductivity to induce bone formation in vivo. - Highlights: • We tried to render protein-based hydrogels suitable for hard tissue regeneration. • We developed a three-component system comprising hydrogel, nano-HAp, and cells. • Nano-HAp significantly improved the mechanical strength of hydrogel. • Encapsulated cells readily elongated and proliferated in 3D cell-laden nanocomposite. • 3D deposition of bone crystals occurred in the hydrogel nanocomposites.

  17. Direct Reprogramming of Human Bone Marrow Stromal Cells into Functional Renal Cells Using Cell-free Extracts

    Directory of Open Access Journals (Sweden)

    Evangelia Papadimou

    2015-04-01

    Full Text Available The application of cell-based therapies in regenerative medicine is gaining recognition. Here, we show that human bone marrow stromal cells (BMSCs, also known as bone-marrow-derived mesenchymal cells, can be reprogrammed into renal proximal tubular-like epithelial cells using cell-free extracts. Streptolysin-O-permeabilized BMSCs exposed to HK2-cell extracts underwent morphological changes—formation of “domes” and tubule-like structures—and acquired epithelial functional properties such as transepithelial-resistance, albumin-binding, and uptake and specific markers E-cadherin and aquaporin-1. Transmission electron microscopy revealed the presence of brush border microvilli and tight intercellular contacts. RNA sequencing showed tubular epithelial transcript abundance and revealed the upregulation of components of the EGFR pathway. Reprogrammed BMSCs integrated into self-forming kidney tissue and formed tubular structures. Reprogrammed BMSCs infused in immunodeficient mice with cisplatin-induced acute kidney injury engrafted into proximal tubuli, reduced renal injury and improved function. Thus, reprogrammed BMSCs are a promising cell resource for future cell therapy.

  18. Rapid, room-temperature synthesis of amorphous selenium/protein composites using Capsicum annuum L extract

    Science.gov (United States)

    Li, Shikuo; Shen, Yuhua; Xie, Anjian; Yu, Xuerong; Zhang, Xiuzhen; Yang, Liangbao; Li, Chuanhao

    2007-10-01

    We describe the formation of amorphous selenium (α-Se)/protein composites using Capsicum annuum L extract to reduce selenium ions (SeO32-) at room temperature. The reaction occurs rapidly and the process is simple and easy to handle. A protein with a molecular weight of 30 kDa extracted from Capsicum annuum L not only reduces the SeO32- ions to Se0, but also controls the nucleation and growth of Se0, and even participates in the formation of α-Se/protein composites. The size and shell thickness of the α-Se/protein composites increases with high Capsicum annuum L extract concentration, and decreases with low reaction solution pH. The results suggest that this eco-friendly, biogenic synthesis strategy could be widely used for preparing inorganic/organic biocomposites. In addition, we also discuss the possible mechanism of the reduction of SeO32- ions by Capsicum annuum L extract.

  19. Comparison of Different Protein Extraction Methods for Gel-Based Proteomic Analysis of Ganoderma spp.

    Science.gov (United States)

    Al-Obaidi, Jameel R; Saidi, Noor Baity; Usuldin, Siti Rokhiyah Ahmad; Hussin, Siti Nahdatul Isnaini Said; Yusoff, Noornabeela Md; Idris, Abu Seman

    2016-04-01

    Ganoderma species are a group of fungi that have the ability to degrade lignin polymers and cause severe diseases such as stem and root rot and can infect economically important plants and perennial crops such as oil palm, especially in tropical countries such as Malaysia. Unfortunately, very little is known about the complex interplay between oil palm and Ganoderma in the pathogenesis of the diseases. Proteomic technologies are simple yet powerful tools in comparing protein profile and have been widely used to study plant-fungus interaction. A critical step to perform a good proteome research is to establish a method that gives the best quality and a wide coverage of total proteins. Despite the availability of various protein extraction protocols from pathogenic fungi in the literature, no single extraction method was found suitable for all types of pathogenic fungi. To develop an optimized protein extraction protocol for 2-DE gel analysis of Ganoderma spp., three previously reported protein extraction protocols were compared: trichloroacetic acid, sucrose and phenol/ammonium acetate in methanol. The third method was found to give the most reproducible gels and highest protein concentration. Using the later method, a total of 10 protein spots (5 from each species) were successfully identified. Hence, the results from this study propose phenol/ammonium acetate in methanol as the most effective protein extraction method for 2-DE proteomic studies of Ganoderma spp. PMID:27016942

  20. Protein expression profile in the differentiation of rat bone marrow stromal cells into Schwann cell-like cells

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    During the last decade,increasing evidence suggested that bone marrow stromal cells(MSCs) have the potential to differentiate into neural lineages.Many studies have reported that MSCs showed morphological changes and expressed a limited number of neural proteins under experimental conditions.However,no proteomic studies on MSCs differentiated into Schwann cell-like cells have been reported.In this study,we isolated MSCs from adult Sprague-Dawley rat femur and tibia bone marrows and induced the cells in vitro under specific conditions.By using two-dimensional gel electrophoresis(2-DE),we compared the protein profiles of MSCs before and after induced differentiation.We obtained 792 protein spots in the protein profile by 2-DE,and found that 74 spots changed significantly before and after the differentiation using PDQuest software,with 43 up-regulated and 31 down-regulated.We analyzed these 74 spots by a matrix assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF-MS) and by database searching,and found that they could be grouped into various classes,including cytoskeleton and structure proteins,growth factors,metabolic proteins,chaperone proteins,receptor proteins,cell cycle proteins,calcium binding proteins,and other proteins.These proteins also include neural and glial proteins,such as BDNF,CNTF and GFAP.The results may provide valuable proteomic information about the differentiation of MSCs into Schwann cell-like cells.

  1. Extraction of Protein Sequence Motif Information using PSO K-Means

    OpenAIRE

    Gowri, R.; Rathipriya, R.

    2015-01-01

    The main objective of the paper is to find the motif information.The functionalities of the proteins are ideally found from their motif information which is extracted using various techniques like clustering with k-means, hybrid k-means, self-organising maps, etc., in the literature. In this work protein sequence information is extracted using optimised k-means algorithm. The particle swarm optimisation technique is one of the frequently used optimisation method. In the current work the PSO k...

  2. The Influence of Chopping Duration on the Degree of Proteins Extraction

    OpenAIRE

    Daniela Ianiţchi; Lucica Nistor; Gratziela Victoria Bahaciu; Camelia Hodoşan; Laura Urdeş; Vasile Băcilă

    2010-01-01

    The quality of meat preparations, in terms of binding compositions, is directly influenced by the degree of extractibility of structural proteins, which in turn is influenced by a number of factors such as quality of raw and auxiliary materials, how various operations are conducted the technological process, type of equipment used, etc. This paper aims to follow how the degree of extractability of sarcoplasmic and myofibril proteins it is influenced by chopping and the type raw material. From...

  3. Ridge preservation with acellular dermal matrix and anorganic bone matrix cell-binding peptide P-15 after tooth extraction in humans. A histologic and morphometric study

    OpenAIRE

    Arthur B. Novaes Jr.; Patricia Garani Fernandes; Flávia Adelino Suaid; Marcio Fernando de Moraes Grisi; Sergio Luis Scombatti de Souza; Mario Taba Jr.; Daniela Bazan Palioto; Valdir Antonio Muglia

    2012-01-01

    Aim: The aim of this study was to analyze by histomorphometric parameters the use of acellular dermal matrix (ADM) with or without anorganic bovine bone matrix (ABM) / synthetic cell-binding peptide P-15 in the formation of bone in human alveoli. Materials and methods: Eighteen patients in need of extraction of maxillary anterior teeth were selected and randomly assigned to the test group (ADM plus ABM/P-15) or the control group (ADM only). Histomorphometric measurements and histological a...

  4. Cold-batter mincing of hot-boned and crust-frozen air-chilled turkey breast allows for reduced sodium content in protein gels.

    Science.gov (United States)

    Lee, H C; Medellin-Lopez, M; Singh, P; Sansawat, T; Chin, K B; Kang, I

    2014-09-01

    The purpose of this research was to evaluate sodium reduction in the protein gels that were prepared with turkey breasts after hot boning (HB), quarter (¼) sectioning, crust-frozen air-chilling (CFAC), and cold temperature mincing. For each of 4 replications, 36 turkeys were slaughtered and eviscerated. One-half of the carcasses were randomly assigned to water immersion chilling for chill boning (CB), whereas the remaining carcasses were immediately HB and quarter-sectioned/crust-frozen air-chilled (HB-¼CFAC) in a freezing room (-12°C, 1.0 m/s). After deboning, CB fillets were conventionally minced, whereas HB-¼CFAC fillets were cold minced up to 27 min with 1 or 2% salt. From the beginning of mincing, the batter temperatures of HB-¼CFAC were lower (P mincing, the batter pH of the HB-¼CFAC (P 0.05) from the pH of CB batters, except for the 1% salt HB-¼CFAC batter after 15 min of mincing. The pattern of pH was not changed when the batters were stored overnight. The protein of 2% salt HB-¼CFAC fillets was more extractable (P mincing technologies appear to improve protein functionality and sodium reduction capacity. PMID:25012854

  5. Controlled trial of the effects of milk basic protein (MBP) supplementation on bone metabolism in healthy adult women.

    Science.gov (United States)

    Aoe, S; Toba, Y; Yamamura, J; Kawakami, H; Yahiro, M; Kumegawa, M; Itabashi, A; Takada, Y

    2001-04-01

    Milk has more beneficial effects on bone health compared to other food sources. Recent in vitro and in vivo studies showed that milk whey protein, especially its basic protein fraction, contains several components capable of both promoting bone formation and inhibiting bone resorption. However, the effects of milk basic protein (MBP) on bone metabolism of humans are not known. The object of this study was to examine the effects of MBP on bone metabolism of healthy adult women. Thirty-three normal healthy women were randomly assigned to treatment with either placebo or MBP (40 mg per day) for six months. The bone mineral density (BMD) of the left calcaneus of each subject was measured at the beginning of the study and after six months of treatment, by dual-energy x-ray absorptiometry. Serum and urine indices of bone metabolism were measured at the base line, three-month intervals, and the end of the study. Daily intake of nutrients was monitored by a three-day food record made at three and six months. The mean (+/- SD) rate of left calcaneus BMD gain of women in the MBP group (3.42 +/- 2.05%) was significantly higher than that of women in the placebo group (2.01 +/- 1.75%, P = 0.042). As compared with the placebo group, urinary cross-linked N-teleopeptides of type-I collagen/creatinine and deoxypyridinoline/creatinine were significantly decreased in the MBP group (p supplementation of 40 mg in healthy adult women can significantly increase their BMD independent of dietary intake of minerals and vitamins. This increase in BMD might be primarily mediated through inhibition of osteoclast-mediated bone resorption by the MBP supplementation. PMID:11388472

  6. Socket Preservation Therapy with Acellular Dermal Matrix and Mineralized Bone Allograft After Tooth Extraction in Humans: A Clinical and Histomorphometric Study.

    Science.gov (United States)

    Fernandes, Patricia Garani; Muglia, Valdir Antonio; Reino, Danilo Maeda; Maia, Luciana Prado; de Moraes Grisi, Marcio Fernando; de Souza, Sergio Luís; Taba, Mario; Palioto, Daniela Bazan; de Almeida, Adriana G; Novaes, Arthur Belém

    2016-01-01

    The aim of this study was to analyze through clinical and histomorphometric parameters the use of acellular dermal matrix (ADM) with or without mineralized bone allograft (AB) on bone formation in human alveoli after a 6- to 8-month healing period. A total of 19 patients in need of extraction of the maxillary anterior teeth were selected and randomly assigned to the test group (ADM plus AB) or to the control group (ADM only). Clinical and histomorphometric measurements and histologic analysis were recorded 6 to 8 months after ridge preservation procedures. Clinical parameters and amount of mineralized and nonmineralized tissue were measured and analyzed. In the clinical measurements, the test group showed reduced bone loss in the buccopalatal dimension after 6 to 8 months (intragroup analysis P acellular dermal matrix in association with mineralized bone allograft reduced alveolar bone loss in the anterior maxillae both in height and width after a follow-up period of 6 to 8 months. PMID:26901306

  7. Effects of deer bone extract on the expression of pro-inflammatory cytokine and cartilage-related genes in monosodium iodoacetate-induced osteoarthritic rats.

    Science.gov (United States)

    Lee, Hyunji; Choi, Hyeon-Son; Park, Yooheon; Ahn, Chang Won; Jung, Sung Ug; Park, Soo Hyun; Suh, Hyung Joo

    2014-01-01

    Deer bone extract has the potential to relieve the discomfort or the articular cartilaginous damage associated with osteoarthritic (OA) and may be useful as a natural supplement for OA treatment without serious side effects. We analyzed the expression of pro-inflammatory cytokine and cartilage-related genes in monosodium iodoacetate-induced OA rats. Increases in the levels of serum pro-inflammatory cytokines, such as interleukin-1β, interleukin-6, and tumor necrosis factor-α were significantly inhibited by the administration of deer bone extract (pCOL2) and tissue inhibitors of metalloproteinases (TIMPs) mRNAs in the cartilage were significantly inhibited by deer bone extract treatment (pCOL2 and TIMP mRNAs and the down-regulation of MMP mRNAs by suppressing the expression of pro-inflammatory cytokine mRNAs. PMID:25273135

  8. Qualitative characterization of the protein fraction of exopolymeric substances (EPS) extracted with EDTA from sludge.

    Science.gov (United States)

    Bourven, Isabelle; Costa, Guy; Guibaud, Gilles

    2012-01-01

    Exopolymeric substances (EPS) were extracted by EDTA from activated and anaerobic granular sludge. Due to the presence of EDTA in EPS extract, interferences were pointed out for the characterization of EPS by means of the colorimetric methods and fluorescence spectroscopy. Other methods have been investigated to characterize the EPS protein fraction. Size exclusion chromatography (SEC), performed at a fluorescence excitation-emission matrix of 221/360 nm (tryptophan protein-like substances) for detection, was suitable and allowed obtaining a fingerprint of the protein-like substance fractions and determining apparent molecular weight (MW). Polyacrylamide gel electrophoresis (PAGE) was performed under either native or denaturing conditions. Various staining applications after EPS migration are effective in obtaining a protein (silver staining) or glycoprotein (PAS staining) fingerprint or MW distribution. SEC and PAGE are both appropriate techniques for the qualitative characterization of protein fractions from EPS extracted by EDTA and moreover differentiate EPS according to sludge origin and type. PMID:22154750

  9. Bone morphogenetic protein 6 polymorphisms are associated with radiographic progression in ankylosing spondylitis.

    Directory of Open Access Journals (Sweden)

    Young Bin Joo

    Full Text Available Nearly 25 genetic loci associated with susceptibility to ankylosing spondylitis (AS have been identified by several large studies. However, there have been limited studies to identify the genes associated with radiographic severity of the disease. Thus we investigated which genes involved in bone formation pathways might be associated with radiographic severity in AS.A total of 417 Korean AS patients were classified into two groups based on the radiographic severity as defined by the modified Stoke' Ankylosing Spondylitis Spinal Score (mSASSS system. Severe AS was defined by the presence of syndesmophytes and/or fusion in the lumbar or cervical spine (n = 195. Mild AS was defined by the absence of any syndesmophyte or fusion (n = 170. A total of 251 single nucleotide polymorphisms (SNPs within 52 genes related to bone formation were selected and genotyped. Odds ratios (OR and 95% confidence interval (95% CI were analysed by multivariate logistic regression controlling for age at onset of symptoms, sex, disease duration, and smoking status as covariates.We identified new loci of bone morphogenetic protein 6 (BMP6 associated with radiographic severity in patients with AS that passed false discovery rate threshold. Two SNPs in BMP6 were significantly associated with radiologic severity [rs270378 (OR 1.97, p = 6.74 × 10(-4 and rs1235192 [OR 1.92, p = 1.17 × 10(-3] adjusted by covariates.This is the first study to demonstrate that BMP6 is associated with radiographic severity in AS, supporting the role wingless-type like/BMP pathway on radiographic progression in AS.

  10. Public awareness of the bone morphogenic protein controversy: Evidence from news publications

    Directory of Open Access Journals (Sweden)

    Doniel Drazin

    2014-01-01

    Full Text Available Background: Use of recombinant human bone morphogenic protein-2 (rhBMP-2 in spinal fusion has seen a tremendous increase. Public awareness of rhBMP-2 and its complications has not been assessed. The authors studied published news media articles to analyze information provided to the public on this bone graft substitute. Methods: We utilized the academic database, LexisNexis, to locate newspaper articles published between January 2001 and July 2013. All articles were coded by a coder and reviewed by the principal investigator. Results: The search identified 87 national and 99 local newspaper articles. Complications mentioned in national newspapers included cancer (24%, retrograde ejaculation (24%, and abnormal bone growth (14%. Local newspapers cited cancer (14%, inflammation (14%, and retrograde ejaculation (9.2% most frequently. Fifty national (59% and 35 local (54% articles had no mention of complications. Sources of evidence cited by articles were (in order of frequency: Governmental agencies, medical research or published studies, healthcare personnel or patients, and companies or corporations. Conclusions: Only a small percentage of newspaper articles presented potential complications. Despite lack of clear scientific causal relationship between rhBMP-2 and cancer, this risk was disproportionately reported. Additionally, many did not cite scientific sources. Lack of reliable information available to the public reiterates the role of physicians in discussing risks and benefits BMP use in spinal surgery, assuring that patients are making informed decisions. Future news media articles should present risks in an impartial and evidence-based manner. Collaboration between advocacy groups, medical institutions, and media outlets would be beneficial in achieving this goal.

  11. Automatic extraction of gene ontology annotation and its correlation with clusters in protein networks

    Directory of Open Access Journals (Sweden)

    Mazo Ilya

    2007-07-01

    Full Text Available Abstract Background Uncovering cellular roles of a protein is a task of tremendous importance and complexity that requires dedicated experimental work as well as often sophisticated data mining and processing tools. Protein functions, often referred to as its annotations, are believed to manifest themselves through topology of the networks of inter-proteins interactions. In particular, there is a growing body of evidence that proteins performing the same function are more likely to interact with each other than with proteins with other functions. However, since functional annotation and protein network topology are often studied separately, the direct relationship between them has not been comprehensively demonstrated. In addition to having the general biological significance, such demonstration would further validate the data extraction and processing methods used to compose protein annotation and protein-protein interactions datasets. Results We developed a method for automatic extraction of protein functional annotation from scientific text based on the Natural Language Processing (NLP technology. For the protein annotation extracted from the entire PubMed, we evaluated the precision and recall rates, and compared the performance of the automatic extraction technology to that of manual curation used in public Gene Ontology (GO annotation. In the second part of our presentation, we reported a large-scale investigation into the correspondence between communities in the literature-based protein networks and GO annotation groups of functionally related proteins. We found a comprehensive two-way match: proteins within biological annotation groups form significantly denser linked network clusters than expected by chance and, conversely, densely linked network communities exhibit a pronounced non-random overlap with GO groups. We also expanded the publicly available GO biological process annotation using the relations extracted by our NLP technology

  12. Enhanced healing of rabbit segmental radius defects with surface-coated calcium phosphate cement/bone morphogenetic protein-2 scaffolds

    International Nuclear Information System (INIS)

    Large osseous defects remain a difficult clinical problem in orthopedic surgery owing to the limited effective therapeutic options, and bone morphogenetic protein-2 (BMP-2) is useful for its potent osteoinductive properties in bone regeneration. Here we build a strategy to achieve prolonged duration time and help inducting new bone formation by using water-soluble polymers as a protective film. In this study, calcium phosphate cement (CPC) scaffolds were prepared as the matrix and combined with sodium carboxymethyl cellulose (CMC-Na), hydroxypropylmethyl cellulose (HPMC), and polyvinyl alcohol (PVA) respectively to protect from the digestion of rhBMP-2. After being implanted in the mouse thigh muscles, the surface-modified composite scaffolds evidently induced ectopic bone formation. In addition, we further evaluated the in vivo effects of surface-modified scaffolds in a rabbit radius critical defect by radiography, three dimensional micro-computed tomographic (μCT) imaging, synchrotron radiation-based micro-computed tomographic (SRμCT) imaging, histological analysis, and biomechanical measurement. The HPMC-modified CPC scaffold was regarded as the best combination for segmental bone regeneration in rabbit radius. - Highlights: • A simple surface-coating method was used to fabricate composite scaffolds. • Growth factor was protected from rapid depletion via superficial coating. • Significant promotion of bone regeneration was achieved. • HPMC-modification displayed optimal effect of bone regeneration

  13. Enhanced healing of rabbit segmental radius defects with surface-coated calcium phosphate cement/bone morphogenetic protein-2 scaffolds

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Yi; Hou, Juan; Yin, ManLi [Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); Wang, Jing, E-mail: biomatwj@163.com [Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); Liu, ChangSheng, E-mail: csliu@sh163.net [Engineering Research Center for Biomedical Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China); The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237 (China); Key Laboratory for Ultrafine Materials of Ministry of Education, East China University of Science and Technology, Shanghai 200237 (China)

    2014-11-01

    Large osseous defects remain a difficult clinical problem in orthopedic surgery owing to the limited effective therapeutic options, and bone morphogenetic protein-2 (BMP-2) is useful for its potent osteoinductive properties in bone regeneration. Here we build a strategy to achieve prolonged duration time and help inducting new bone formation by using water-soluble polymers as a protective film. In this study, calcium phosphate cement (CPC) scaffolds were prepared as the matrix and combined with sodium carboxymethyl cellulose (CMC-Na), hydroxypropylmethyl cellulose (HPMC), and polyvinyl alcohol (PVA) respectively to protect from the digestion of rhBMP-2. After being implanted in the mouse thigh muscles, the surface-modified composite scaffolds evidently induced ectopic bone formation. In addition, we further evaluated the in vivo effects of surface-modified scaffolds in a rabbit radius critical defect by radiography, three dimensional micro-computed tomographic (μCT) imaging, synchrotron radiation-based micro-computed tomographic (SRμCT) imaging, histological analysis, and biomechanical measurement. The HPMC-modified CPC scaffold was regarded as the best combination for segmental bone regeneration in rabbit radius. - Highlights: • A simple surface-coating method was used to fabricate composite scaffolds. • Growth factor was protected from rapid depletion via superficial coating. • Significant promotion of bone regeneration was achieved. • HPMC-modification displayed optimal effect of bone regeneration.

  14. An Approach to Protein Name Extraction Using Heuristics and a Dictionary.

    Science.gov (United States)

    Seki, Kazuhiro; Mostafa, Javed

    2003-01-01

    Proposes a method for protein name extraction from biological texts. The method exploits hand-crafted rules based on heuristics and a set of protein names (dictionary). The approach avoids use of natural language processing tools so as to improve processing speed. Evaluation experiments were conducted in terms of: accuracy, generalizability, and…

  15. Umami taste amino acids produced by hydrolyzing extracted protein from tomato seed meal

    Science.gov (United States)

    Enzymatic hydrolysis was performed for extracting protein to prepare umami taste amino acids from defatted tomato seed meal (DTSM) which is a by-product of tomato processing. Papain was used as an enzyme for the hydrolysis of DTSM. The particle size distribution of DTSM, protein concentration and fr...

  16. Plasma Surface Modification for Immobilization of Bone Morphogenic Protein-2 on Polycaprolactone Scaffolds

    Science.gov (United States)

    Kim, Byung Hoon; Myung, Sung Woon; Jung, Sang Chul; Ko, Yeong Mu

    2013-11-01

    The immobilization of recombinant human bone formation protein-2 (rhBMP-2) on polycaprolactone (PCL) scaffolds was performed by plasma polymerization. RhBMP-2, which induces osteoblast differentiation in various cell types, is a growth factor that plays an important role in bone formation and repair. The surface of the PCL scaffold was functionalized with the carboxyl groups of plasma-polymerized acrylic acid (PPAA) thin films. Plasma polymerization was carried out at a discharge power of 60 W at an acrylic acid flow rate of 7 sccm for 5 min. The PPAA thin film exhibited moderate hydrophilic properties and possessed a high density of carboxyl groups. Carboxyl groups and rhBMP-2 on the PCL scaffolds surface were identified by attenuated total reflection Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy, respectively. The alkaline phosphatase activity assay showed that the rhBMP-2 immobilized PCL scaffold increased the level of MG-63 cell differentiation. Plasma surface modification for the preparation of biomaterials, such as biofunctionalized polymer scaffolds, can be used for the binding of bioactive molecules in tissue engineering.

  17. Comparison and optimization of methods for the simultaneous extraction of DNA, RNA, proteins, and metabolites.

    Science.gov (United States)

    Vorreiter, Fränze; Richter, Silke; Peter, Michel; Baumann, Sven; von Bergen, Martin; Tomm, Janina M

    2016-09-01

    The challenge of performing a time-resolved comprehensive analysis of molecular systems has led to the quest to optimize extraction methods. When the size of a biological sample is limited, there is demand for the simultaneous extraction of molecules representing the four areas of "omics": genomics, transcriptomics, proteomics, and metabolomics. Here we optimized a protocol for the simultaneous extraction of DNA, RNA, proteins, and metabolites and compared it with two existing protocols. Our optimization comprised the addition of a methanol/chloroform metabolite purification before the separation of DNA/RNA and proteins. Extracted DNA, RNA, proteins, and metabolites were quantitatively and/or qualitatively analyzed. Of the three methods, only the newly developed protocol yielded all biomolecule classes of adequate quantity and quality. PMID:27237373

  18. Three-Dimensional Upper Lip and Nostril Sill Changes After Cleft Alveolus Reconstruction Using Autologous Bone Grafting Versus Recombinant Human Bone Morphogenetic Protein-2.

    Science.gov (United States)

    Raposo-Amaral, Cassio Eduardo; Denadai, Rafael; Alonso, Nivaldo

    2016-06-01

    Cleft alveolus in patients with unilateral complete cleft lip and palate has been alternatively reconstructed with recombinant human bone morphogenetic protein (rhBMP)-2. However, its effects on upper lip and nostril sill anatomy are not known. Thus, the objective of this investigation was to assess and compare upper lip and nostril sill changes after cleft alveolus reconstruction with autologous bone from the iliac crest region and rhBMP-2. Patients were randomly allocated into 2 groups. In group 1, autologous bone from the iliac crest region was used to fill the cleft alveolus (n = 4), and in group 2, rhBMP-2 was used to fill the cleft alveolus (n = 8). Preoperatively and at one after the surgery, computerized tomography (CT) was performed. Reformatted CT imaging was used to perform cephalometric linear measurements of the upper lip and nostril sill regions. Inter- and intragroup data of the pre and postoperative reformatted CT measurements of the upper lip and nostril sill regions did not show differences (P >0.05) in cutaneous upper lip height and projection, nostril sill elevation, and subnasale projection. There were no significant upper lip and nostril sill anatomical changes after cleft alveolus reconstruction using autologous bone grafting and rhBMP-2. PMID:27244210

  19. Testosterone delivered with a scaffold is as effective as bone morphologic protein-2 in promoting the repair of critical-size segmental defect of femoral bone in mice.

    Directory of Open Access Journals (Sweden)

    Bi-Hua Cheng

    Full Text Available Loss of large bone segments due to fracture resulting from trauma or tumor removal is a common clinical problem. The goal of this study was to evaluate the use of scaffolds containing testosterone, bone morphogenetic protein-2 (BMP-2, or a combination of both for treatment of critical-size segmental bone defects in mice. A 2.5-mm wide osteotomy was created on the left femur of wildtype and androgen receptor knockout (ARKO mice. Testosterone, BMP-2, or both were delivered locally using a scaffold that bridged the fracture. Results of X-ray imaging showed that in both wildtype and ARKO mice, BMP-2 treatment induced callus formation within 14 days after initiation of the treatment. Testosterone treatment also induced callus formation within 14 days in wildtype but not in ARKO mice. Micro-computed tomography and histological examinations revealed that testosterone treatment caused similar degrees of callus formation as BMP-2 treatment in wildtype mice, but had no such effect in ARKO mice, suggesting that the androgen receptor is required for testosterone to initiate fracture healing. These results demonstrate that testosterone is as effective as BMP-2 in promoting the healing of critical-size segmental defects and that combination therapy with testosterone and BMP-2 is superior to single therapy. Results of this study may provide a foundation to develop a cost effective and efficient therapeutic modality for treatment of bone fractures with segmental defects.

  20. Study on preventive and therapeutic effect of Chinese medicinal herbal extracts on rat with bone marrow injury induced by radiation exposure

    International Nuclear Information System (INIS)

    Objective: To examine the effect of Chinese medicinal herbal extracts, Danggui (Radix angelicae sinensis), Chuanxiong (Rhizoma chuanxiong), Huangqi (Radix astragali), and Danshen (Radix salviae miltiorrhizae) on rats with bone marrow injury induced with whole-body gamma-ray exposure. Methods: Sixty male rats were randomly divided into three groups, control group, model group (irradiation only with no administration of the extracts), and drug treatment group (irradiation and administration of Chinese medicinal herbal extracts). Rats were irradiated with 6 Gy cobolt-60 gamma rays after administration of the extracts for two weeks. The number of marrow nucleate cells was counted, and VEGF and PDGF expression were measured with Western blot method on the 7th day since the irradiation. Results: Bone marrow nucleate cells and VEGF and PDGF expression in bone marrow cells in the model group were significantly lower than those in the control group (P<0.01), and these values in the drug treatment group were significantly higher than those in the model group (P<0.01 or P<0.05). Conclusion: The extracts of Chuanxiong, Danggui, Huangqi, and Danshen can be used to prevent from ration bone marrow injury in rats. (authors)

  1. The combined mechanism of bone morphogenetic protein- and calcium phosphate-induced skeletal tissue formation by human periosteum derived cells.

    Science.gov (United States)

    Bolander, J; Ji, W; Geris, L; Bloemen, V; Chai, Y C; Schrooten, J; Luyten, F P

    2016-01-01

    When combining osteogenic progenitor cells such as human periosteum derived cells (hPDCs) with osteoconductive biomaterials like calcium phosphate (CaP)-scaffolds, in vivo bone formation can be achieved. This process is dependent on the early activation of Bone morphogenetic protein (BMP)-signalling. However, the bone forming process is slow and routinely only a limited amount of bone and bone marrow is formed. Therefore, we hypothesised that a robust clinically relevant outcome could be achieved by adding more physiological levels of potent BMP-ligands to these cell- and CaP-based constructs. For this, hPDCs were characterised for their responsiveness to BMP-ligands upon in vitro 2D stimulation. BMP-2, -4, -6 and -9 robustly induced osteochondrogenic differentiation. Subsequently, these ligands were coated onto clinically approved CaP-scaffolds, BioOss® and CopiOs®, followed by hPDC-seeding. Protein lysates and conditioned media were investigated for activation of BMP signalling pathways. Upon in vivo implantation, the most abundant bone formation was found in BMP-2 and BMP-6-coated scaffolds. Implanted cells actively contributed to the newly formed bone. Remnants of cartilage could be observed in BMP-coated CopiOs®-constructs. Computational analysis displayed that the type of BMP-ligand as well as the CaP-scaffold affects skeletal tissue formation, observed in a qualitative as well as quantitative manner. Furthermore, the in vitro mechanism appears to predict the in vivo outcome. This study presents further evidence for the potential of BMP-technology in the development of clinically relevant cell-based constructs for bone regenerative strategies. PMID:26728496

  2. How Does Alkali Aid Protein Extraction in Green Tea Leaf Residue: A Basis for Integrated Biorefinery of Leaves.

    Directory of Open Access Journals (Sweden)

    Chen Zhang

    Full Text Available Leaf protein can be obtained cost-efficiently by alkaline extraction, but overuse of chemicals and low quality of (denatured protein limits its application. The research objective was to investigate how alkali aids protein extraction of green tea leaf residue, and use these results for further improvements in alkaline protein biorefinery. Protein extraction yield was studied for correlation to morphology of leaf tissue structure, protein solubility and hydrolysis degree, and yields of non-protein components obtained at various conditions. Alkaline protein extraction was not facilitated by increased solubility or hydrolysis of protein, but positively correlated to leaf tissue disruption. HG pectin, RGII pectin, and organic acids were extracted before protein extraction, which was followed by the extraction of cellulose and hemi-cellulose. RGI pectin and lignin were both linear to protein yield. The yields of these two components were 80% and 25% respectively when 95% protein was extracted, which indicated that RGI pectin is more likely to be the key limitation to leaf protein extraction. An integrated biorefinery was designed based on these results.

  3. Osseointegration by bone morphogenetic protein-2 and transforming growth factor beta2 coated titanium implants in femora of New Zealand white rabbits

    OpenAIRE

    Fritz Thorey; Henning Menzel; Corinna Lorenz; Gerhard Gross; Andrea Hoffmann; Henning Windhagen

    2011-01-01

    Background: Intramembranous bone formation is essential in uncemented joint replacement to provide a mechanical anchorage of the implant. Since the discovery of bone morphogenic proteins (BMPs) by Urist in 1965, many studies have been conducted to show the influence of growth factors on implant ingrowth. In this study, the influence of bone morphogenetic protein-2 (rhBMP-2) and transforming growth factor β2 (TGF-β2) on implant osseointegration was investigated. Materials and Methods: Thir...

  4. An approach to improve kernel-based Protein-Protein Interaction extraction by learning from large-scale network data.

    Science.gov (United States)

    Li, Lishuang; Guo, Rui; Jiang, Zhenchao; Huang, Degen

    2015-07-15

    Protein-Protein Interaction extraction (PPIe) from biomedical literatures is an important task in biomedical text mining and has achieved desirable results on the annotated datasets. However, the traditional machine learning methods on PPIe suffer badly from vocabulary gap and data sparseness, which weakens classification performance. In this work, an approach capturing external information from the web-based data is introduced to address these problems and boost the existing methods. The approach involves three kinds of word representation techniques: distributed representation, vector clustering and Brown clusters. Experimental results show that our method outperforms the state-of-the-art methods on five publicly available corpora. Our code and data are available at: http://chaoslog.com/improving-kernel-based-protein-protein-interaction-extraction-by-unsupervised-word-representation-codes-and-data.html. PMID:25864936

  5. Alcohol-extracted, but not intact, dietary soy protein lowers lipoprotein(a) markedly

    DEFF Research Database (Denmark)

    Meinertz, Hans; Nilausen, Karin; Hilden, Jørgen

    2002-01-01

    , liquid-formula diets containing casein, soy protein, or alcohol-extracted soy protein. Dietary periods of 32 days were separated by washout periods on self-selected diets. Fasting lipid and Lp(a) levels were measured throughout. Median Lp(a) concentration was >2-fold greater after 28 to 32 days on a soy......% after 1 week (P<0.01) and subsequently decreased toward baseline; extracted soy protein and casein diets never exhibited increased median Lp(a) levels, and after 28 to 32 days, these levels were decreased >60% below baseline (P<0.001 and P<0.01, respectively). Low density lipoprotein cholesterol...

  6. Nucleic acid and protein extraction from electropermeabilized E. coli cells on a microfluidic chip

    DEFF Research Database (Denmark)

    Matos, T.; Senkbeil, Silja; Mendonça, A.;

    2013-01-01

    Due to the extensive use of nucleic acid and protein analysis of bacterial samples, there is a need for simple and rapid extraction protocols for both plasmid DNA and RNA molecules as well as reporter proteins like the green fluorescent protein (GFP). In this report, an electropermeability...... can be avoided and the transiently formed pores can be closed again and the cells survive. This method has been used to extract RNA and GFP molecules under conditions of electropermeability. Plasmid DNA could be recovered when the applied voltage was increased to 2 V, thus causing complete cell lysis....

  7. Alkaline Extraction of Cobia (Rachycentroncanadum) Proteins: Physicochemical characteristics, functional and thermal properties

    OpenAIRE

    Sabrina de La Rocha; Jorge Alberto V. Costa

    2015-01-01

    Cobia (Rachycentroncanadum) is an important emerging species in Brazilian mariculture. The aim of the study was to obtain and characterize a protein isolate from cobia muscle using chemical extraction process by alkaline solubilization and isoelectric precipitation of proteins. The extraction yield was 98.17g/100g protein on a dry basis. The highest solubility and water holding capacity (WHC) of cobiaprotein isolate (CPI) was obtained at pH 11and 21.9mL of water per gram of protei...

  8. The Herpes Simplex Virus vhs Protein Induces Endoribonucleolytic Cleavage of Target RNAs in Cell Extracts

    OpenAIRE

    Elgadi, Mabrouk M.; Hayes, Christopher E.; Smiley, James R.

    1999-01-01

    The herpes simplex virus virion host shutoff (vhs) protein (UL41 gene product) is a component of the HSV virion tegument that triggers shutoff of host protein synthesis and accelerated mRNA degradation during the early stages of HSV infection. Previous studies have demonstrated that extracts from HSV-infected cells and partially purified HSV virions display vhs-dependent RNase activity and that vhs is sufficient to trigger accelerated RNA degradation when expressed as the only HSV protein in ...

  9. Ectopic bone formation cannot occur by hydroxyapatite/β-tricalcium phosphate bioceramics in green fluorescent protein chimeric mice

    International Nuclear Information System (INIS)

    Highlights: ► Firstly, chimeric mouse model could be established successfully by bone marrow transplantation after irradiation. ► Secondly, bone induction can occur in wild-type mice 90 days after implantation, but not occur in chimeric mice. ► Thirdly, destruction of immune function will block osteoinduction by calcium phosphate ceramics. - Abstract: Many studies have shown that calcium phosphate ceramics (CP) have osteoconductive and osteoinductive properties; however, the exact mechanism of bone induction has not yet been reported. This study was performed to investigate if destroying immunological function will influence osteogenesis, to explain the mechanism which is unclear. In this study, twenty C57BL/6 mice were divided into two groups (n = 10), in group 1, a hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) ceramic was implanted into both the left and right leg muscles of each mouse; in group 2, ten mice experienced lethal irradiation, then were injected bone marrow (BM) cells from green fluorescent protein (GFP) transgenic mice by tail veil, after bone marrow transplantation (BMT), heart, liver, spleen, lung, kidney, and muscle were harvested for biological analysis, after the GFP chimera model was established successfully, the same HA/β-TCP ceramic was implanted into both leg muscles of each mouse immediately after irradiation. 45 and 90 days after implantation, the ceramics of the two groups were harvested to perform with hematoxylin and eosin (HE) and immunohistochemistry (IHC) staining; the results showed that there was no bone formation in group 2, while new bone tissues were detected in group 1. Our findings suggest that the BM cell from GFP transgenic mice is a good biomarker and it could set a good platform for chimera model; it also shows that BM cell is one of cell resources of bone induction, and destruction of immune function will impede osteoinduction by CP. Overall, our results may shed light on clear mechanism study of bone

  10. Ectopic bone formation cannot occur by hydroxyapatite/{beta}-tricalcium phosphate bioceramics in green fluorescent protein chimeric mice

    Energy Technology Data Exchange (ETDEWEB)

    Cheng Lijia [Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu (China); Duan Xin [Department of Orthopaedics, Chengdu Second People' s Hospital, Chengdu (China); Department of Orthopaedics, West China Hospital, Sichuan University, Chengdu (China); Xiang Zhou [Department of Orthopaedics, West China Hospital, Sichuan University, Chengdu (China); Shi Yujun; Lu Xiaofeng; Ye Feng [Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu (China); Bu Hong, E-mail: hongbu@scu.edu.cn [Key Laboratory of Transplant Engineering and Immunology, Ministry of Health, West China Hospital, Sichuan University, Chengdu (China); Department of Pathology, West China Hospital, Sichuan University, Chengdu (China)

    2012-12-01

    Highlights: Black-Right-Pointing-Pointer Firstly, chimeric mouse model could be established successfully by bone marrow transplantation after irradiation. Black-Right-Pointing-Pointer Secondly, bone induction can occur in wild-type mice 90 days after implantation, but not occur in chimeric mice. Black-Right-Pointing-Pointer Thirdly, destruction of immune function will block osteoinduction by calcium phosphate ceramics. - Abstract: Many studies have shown that calcium phosphate ceramics (CP) have osteoconductive and osteoinductive properties; however, the exact mechanism of bone induction has not yet been reported. This study was performed to investigate if destroying immunological function will influence osteogenesis, to explain the mechanism which is unclear. In this study, twenty C57BL/6 mice were divided into two groups (n = 10), in group 1, a hydroxyapatite/{beta}-tricalcium phosphate (HA/{beta}-TCP) ceramic was implanted into both the left and right leg muscles of each mouse; in group 2, ten mice experienced lethal irradiation, then were injected bone marrow (BM) cells from green fluorescent protein (GFP) transgenic mice by tail veil, after bone marrow transplantation (BMT), heart, liver, spleen, lung, kidney, and muscle were harvested for biological analysis, after the GFP chimera model was established successfully, the same HA/{beta}-TCP ceramic was implanted into both leg muscles of each mouse immediately after irradiation. 45 and 90 days after implantation, the ceramics of the two groups were harvested to perform with hematoxylin and eosin (HE) and immunohistochemistry (IHC) staining; the results showed that there was no bone formation in group 2, while new bone tissues were detected in group 1. Our findings suggest that the BM cell from GFP transgenic mice is a good biomarker and it could set a good platform for chimera model; it also shows that BM cell is one of cell resources of bone induction, and destruction of immune function will impede

  11. Comparison of three detergent-free protein extraction protocols for white adipose tissue.

    Science.gov (United States)

    Sajic, Tatjana; Hopfgartner, Gérard; Szanto, Ildiko; Varesio, Emmanuel

    2011-08-15

    A comparative study of three detergent-free protein extraction protocols--a differential centrifugal fractionation, a delipidation protocol based on the Bligh and Dyer method, and the trifluoroethanol addition as cosolvent to an aqueous buffer--was performed on white adipose tissue. The performance of the protocols directly compatible with liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) was evaluated based on the total protein extraction yield and the protein recovery from different functional and cellular compartments. The most suitable method for the extraction of white adipose tissue proteins from a wide range of cellular and structural compartments was the delipidation protocol based on the Bligh and Dyer method. PMID:21565151

  12. Impact of Ellagic Acid in Bone Formation after Tooth Extraction: An Experimental Study on Diabetic Rats

    Directory of Open Access Journals (Sweden)

    Mazen M. Jamil Al-Obaidi

    2014-01-01

    Full Text Available Objectives. To estimate the impact of ellagic acid (EA towards healing tooth socket in diabetic animals, after tooth extraction. Methods. Twenty-four Sprague Dawley male rats weighing 250–300 g were selected for this study. All animals were intraperitoneally injected with 45 mg/kg (b.w. of freshly prepared streptozotocin (STZ, to induce diabetic mellitus. Then, the animals were anesthetized, and the upper left central incisor was extracted and the whole extracted sockets were filled with Rosuvastatin (RSV. The rats were separated into three groups, comprising 8 rats each. The first group was considered as normal control group and orally treated with normal saline. The second group was regarded as diabetic control group and orally treated with normal saline, whereas the third group comprised diabetic rats, administrated with EA (50 mg/kg orally. The maxilla tissue stained by eosin and hematoxylin (H&E was used for histological examinations and immunohistochemical technique. Fibroblast growth factor (FGF-2 and alkaline phosphatase (ALP were used to evaluate the healing process in the extracted tooth socket by immunohistochemistry test. Results. The reactions of immunohistochemistry for FGF-2 and ALP presented stronger expression, predominantly in EA treated diabetic rat, than the untreated diabetic rat. Conclusion. These findings suggest that the administration of EA combined with RSV may have accelerated the healing process of the tooth socket of diabetic rats, after tooth extraction.

  13. Petroleum ether extract of Cissus quadrangularis (LINN) stimulates the growth of fetal bone during intra uterine developmental period: a morphometric analysis

    OpenAIRE

    Bhagath Kumar Potu; Rao, Muddanna S.; N. Gopalan Kutty; Kumar MR Bhat; Mallikarjuna Rao Chamallamudi; Soubhagya Ranjan Nayak

    2008-01-01

    OBJECTIVE: The aim of the present study was to analyze the effect Cissus quadrangularis plant petroleum ether extract on the development of long bones during the intra-uterine developmental stage in rats. METHODS: Pregnant rats (n=12) were randomly assigned into either a control group (n=6) or a Cissus quadrangularis treatment (n=6) group. Pregnant rats in the Cissus quadrangularis group were treated with Cissus quadrangularis petroleum ether extract at a dose of 500 mg/kg body weight from ge...

  14. PPI-IRO: A two-stage method for protein-protein interaction extraction based on interaction relation ontology

    KAUST Repository

    Li, Chuanxi

    2014-01-01

    Mining Protein-Protein Interactions (PPIs) from the fast-growing biomedical literature resources has been proven as an effective approach for the identifi cation of biological regulatory networks. This paper presents a novel method based on the idea of Interaction Relation Ontology (IRO), which specifi es and organises words of various proteins interaction relationships. Our method is a two-stage PPI extraction method. At fi rst, IRO is applied in a binary classifi er to determine whether sentences contain a relation or not. Then, IRO is taken to guide PPI extraction by building sentence dependency parse tree. Comprehensive and quantitative evaluations and detailed analyses are used to demonstrate the signifi cant performance of IRO on relation sentences classifi cation and PPI extraction. Our PPI extraction method yielded a recall of around 80% and 90% and an F1 of around 54% and 66% on corpora of AIMed and Bioinfer, respectively, which are superior to most existing extraction methods. Copyright © 2014 Inderscience Enterprises Ltd.

  15. Selective separation and enrichment of proteins in aqueous two-phase extraction system

    Institute of Scientific and Technical Information of China (English)

    Feng Qu; Hao Qin; Min Dong; Dong Xu Zhao; Xin Ying Zhao; Jing Hua Zhang

    2009-01-01

    A simple aqueous two-phase extraction system(ATPS)of PEG/phosphate was proposed for selective separation and enrichment of proteins.The combination of ATPE with HPLC was applied to identify the partition of proteins in two phases.Five proteins (bovine serum albumin,Cytochrome C,lysozyme,myoglobin,and trypsin)were used as model proteins to study the effect of phosphate concentration and pH on proteins partition.The PEG/phosphate system was firstly applied to real human saliva and plasma samples,some proteins showed obviously different partition in two phases.The primary results manifest the selective separation and enrichment of proteins in ATPS provided the potential for high abundance proteins depletion in proteomics.

  16. 带血供肌瓣作为骨形态发生蛋白载体修复骨缺损的实验研究%Vascular muscle flap combined with bone morphogenetic protein for forming bone bridge to repair bone defect: experimental study

    Institute of Scientific and Technical Information of China (English)

    裴国献; 杨润功; 魏宽海; 金丹

    2001-01-01

    Objective To investigate the effect of vascular muscle flap as a carrier of bone morphogenetic protein (BMP) to repair long bone defect. Methods Vascular muscle flap with BMP and BMP alone were implanted into the experimental models. Their conditions of new bone formation were observed and compared. Additionally, bone defects were divided into 4 groups in random and repaired respectively with the vascular muscle flap combined with FS/BMP (group A), vascular muscle flap/BMP (group B), bloodless muscle flap/BMP (group C), and autolyzed antigen-extracted allogeneic bone (AAA)/BMP (group D). Their abilities of bone forming were observed. Results In the group of vascular muscle flap combined with BMP, a large amount of cartilage was formed in the gaps of muscles by 3 weeks. The cartilage was absorbed and replaced by normal bone containing hematopoietic bone marrow by 6 weeks with obvious muscle cell atrophy. The wet bone weight of the new bone was (253.52±20.63) mg,which was significantly larger than that of the control group (172.22±13.95) mg (P<0.01).In group A,the cartilage formed by 3 weeks and woven bone formed by 6 weeks;the haversion system formed and muscle cells disappeared by 9 weeks.Natural bone was found and the Tmax measured with torsion test was (0.82±0.04) N*m.The calcium content was (174.55±5.11) μg/g by 12 weeks.The ability of new bone formation in the 4 groups was in the following order: group A was similar to group D, group A>group B>group C. Conclusions Vascular muscle flap can serve as an effective carrier for BMP. Vascular muscle flap combined with FS as carrier is better than vascular muscle flap as a carrier alone.%目的探讨带血供肌瓣作为骨形态发生蛋白(BMP)载体修复骨缺损的可行性。方法观察带血供肌瓣复合BMP和单纯BMP组修复骨缺损时的成骨情况;对纤维蛋白粘合剂、带血供肌瓣、无血运肌瓣、同种异体脱钙骨4种不同BMP载体的成骨能力进行

  17. Magnetic solid-phase extraction of protein with deep eutectic solvent immobilized magnetic graphene oxide nanoparticles.

    Science.gov (United States)

    Xu, Kaijia; Wang, Yuzhi; Ding, Xueqin; Huang, Yanhua; Li, Na; Wen, Qian

    2016-02-01

    As a new type of green solvent, four kinds of choline chloride (ChCl)-based deep eutectic solvents (DESs) have been synthesized, and then a core-shell structure magnetic graphene oxide (Fe3O4-NH2@GO) nanoparticles have been prepared and coated with the ChCl-based DESs. Magnetic solid-phase extraction (MSPE) based Fe3O4-NH2@GO@DES was studied for the first time for the extraction of proteins. The characteristic results of vibrating sample magnetometer (VSM), X-ray diffraction (XRD), Fourier transform infrared spectrometry (FT-IR), thermal gravimetric analysis (TGA) and field emission scanning electron microscopy (FESEM) indicated the successful preparation of Fe3O4-NH2@GO@DES. The concentrations of proteins in studies were determined by a UV-vis spectrophotometer. The advantages of Fe3O4-NH2@GO@DES in protein extraction were compared with Fe3O4-NH2@GO and Fe3O4-NH2, and Fe3O4-NH2@GO@ChCl-glycerol was selected as the suitable extraction solvent. The influence factors of the extraction process such as the pH value, the temperature, the extraction time, the concentration of protein and the amount of Fe3O4-NH2@GO@ChCl-glycerol were evaluated. Desorption experimental result showed 98.73% of BSA could be eluted from the solid extractant with 0.1 mol/L Na2HPO4 solution contained 1 mol/L NaCl. Besides, the conformation of BSA was not changed during the elution by the investigation of circular dichromism (CD) spectra. Furthermore, the analysis of real sample demonstrated that the prepared magnetic nanoparticles did have extraction ability on proteins in bovine whole blood. PMID:26653436

  18. KARAKTERISASI FISIKOKIMIA NANOKALSIUM HASIL EKSTRAKSI TULANG IKAN NILA MENGGUNAKAN LARUTAN BASA DAN ASAM [Physicochemical Characterization of Nano Calcium from Tilapia Bone Extracted by Alkaline and Acid Solution

    Directory of Open Access Journals (Sweden)

    Vanessa Lekahena*

    2014-06-01

    Full Text Available The utilization of tilapia (Oreochromis niloticus bones as a source of natural calcium was done by alkaline (NaOH and acid (HCl extraction. The extraction process aims to soften the bones are milled using disc mill into nano sized calcium powder or nanocalcium. The objective of this research was to study the physicochemical properties of nanocalcium obtained from alkaline and acid extraction of tilapia bones. The results of physicochemical analysis properties of nanocalcium sample showed that nanocalcium extracted by alkaline solution had better properties than that of the untreated sample and nanocalcium extracted by acid solution. The results was indicated by the higher yield, brighter colors, smaller particle size, and calcium content (20.67% and phosphorus (10.09% with a ratio of Ca/P = 2.0. The Fourier Transform Infrared (FTIR spectra profile indicated the presence of phosphate group (PO43- at the bands of 469, 563, 603, 961 and 1035 cm-1, and the presence of apatite carbonate group (CO32- was indicated by intense bands at 873, 1416, 1456 and 1563 cm-1. The morphology of the sampel surface of the alkaline extracted sample was smoother, denser, and the grain size formed were relatively larger. The formed crystalline phases were HAp (hydroxyapatite, carbonate apatite type A [Ca10(PO46CO3] and B [Ca10(PO43(CO33(OH2] with crystalline degree of 78.4%.

  19. Bone morphogenetic protein 7 induces cementogenic differentiation of human periodontal ligament-derived mesenchymal stem cells.

    Science.gov (United States)

    Torii, D; Tsutsui, T W; Watanabe, N; Konishi, K

    2016-01-01

    Bone morphogenetic protein 7 (BMP-7) is a multifunctional differentiation factor that belongs to the transforming growth factor superfamily. BMP-7 induces gene expression of protein tyrosine phosphatase-like, member A/cementum attachment protein (PTPLA/CAP) and cementum protein 1 (CEMP1), both of which are markers of cementoblasts and cementocytes. In the previous study, we reported that BMP-7 treatment enhanced PTPLA/CAP and CEMP1 expression in both normal and immortal human periodontal ligament (PDL) cells. To elucidate the molecular mechanisms of the gene expression of these molecules, in this study, we identified a functional transcription activator binding region in the promoter region of PTPLA/CAP and CEMP1 that is responsive to BMP signals. Here, we report that some short motifs termed GC-rich Smad-binding elements (GC-SBEs) that are located in the human PTPLA/CAP promoter and CEMP1 promoter are BMP-7 responsive as analyzed with luciferase promoter assays. On the other hand, we found that transcription of Sp7/Osterix and PTPLA/CAP was up-regulated after 1 week of BMP-7 treatment on purified normal human PDL cells as a result of gene expression microarray analysis. Furthermore, transcription of Sp7/Osterix, runt-related transcription factor 2 (RUNX2), and alkaline phosphatase (ALP) was up-regulated after 2 weeks of BMP-7 treatment, whereas gene expression of osteo/odontogenic markers such as integrin-binding sialoprotein (IBSP), collagen, type I, alpha 1 (COL1A1), dentin matrix acidic phosphoprotein 1 (DMP1), and dentin sialophosphoprotein (DSPP) was not up-regulated in purified normal or immortal human PDL cells as a result of qRT-PCR. The results suggest that BMP-7 mediates cementogenesis via GC-SBEs in human PDL cells and that its molecular mechanism is different from that for osteo/odontogenesis. PMID:25464857

  20. Identification of bone morphogenetic protein 9 (BMP9) as a novel profibrotic factor in vitro.

    Science.gov (United States)

    Muñoz-Félix, José M; Cuesta, Cristina; Perretta-Tejedor, Nuria; Subileau, Mariela; López-Hernández, Francisco J; López-Novoa, José M; Martínez-Salgado, Carlos

    2016-09-01

    Upregulated synthesis of extracellular matrix (ECM) proteins by myofibroblasts is a common phenomenon in the development of fibrosis. Although the role of TGF-β in fibrosis development has been extensively studied, the involvement of other members of this superfamily of cytokines, the bone morphogenetic proteins (BMPs) in organ fibrosis has given contradictory results. BMP9 is the main ligand for activin receptor-like kinase-1 (ALK1) TGF-β1 type I receptor and its effect on fibrosis development is unknown. Our purpose was to study the effect of BMP9 in ECM protein synthesis in fibroblasts, as well as the involved receptors and signaling pathways. In cultured mice fibroblasts, BMP9 induces an increase in collagen, fibronectin and connective tissue growth factor expression, associated with Smad1/5/8, Smad2/3 and Erk1/2 activation. ALK5 inhibition with SB431542 or ALK1/2/3/6 with dorsomorphin-1, inhibition of Smad3 activation with SIS3, and inhibition of the MAPK/Erk1/2 with U0126, demonstrates the involvement of these pathways in BMP9-induced ECM synthesis in MEFs. Whereas BMP9 induced Smad1/5/8 phosphorylation through ALK1, it also induces Smad2/3 phosphorylation through ALK5 but only in the presence of ALK1. Summarizing, this is the first study that accurately identifies BMP9 as a profibrotic factor in fibroblasts that promotes ECM protein expression through ALK1 and ALK5 receptors. PMID:27208502

  1. Extraction of rice bran protein concentrate and its application in bread

    Directory of Open Access Journals (Sweden)

    W. James Harper

    2005-01-01

    Full Text Available In a study of rice bran protein concentrate (RBPC preparation, the alkaline extraction was performed. The objectives of this study were to determine the optimal extracting conditions of RBPC and apply RBPC into a bread recipe in order to improve bread quality and make acceptable product to consumers. The design used in extraction was a central composite design. The response surface methodology was chosen to graphically express the relationship between pH and extraction time with the output variables of protein content and percent yield of RBPC. It was found that optimal extracting conditions were pH 11 and 45 min., resulting in 69.16% protein content and 8.06% yield of RBPC. When incorporating 1-5% of RBPC in abread recipe, the weight loss and microbial counts of breads were decreased compared to those of control bread. The higher protein content and fiber in bread was corresponding to the amount of RBPC added. Therefore, adding RBPC can significantly increase protein and fiber content in bread. However, results from sensory evaluation showed that adding more than 1% of RBPC decreased the liking scores of color, taste, odor, texture, and overall liking. The results of this study could be used as a basic knowledge of RBPC utilization in other food products.

  2. Influence of dietary protein, energy and corticosteroids on protein turnover, proteoglycan sulphation and growth of long bone and skeletal muscle in the rat.

    Science.gov (United States)

    Yahya, Z A; Tirapegui, J O; Bates, P C; Millward, D J

    1994-11-01

    1. We report here the extent to which changes in protein turnover contribute to the previously described inhibition of growth of rat tibial length and skeletal muscle mass in response to protein deficiency [1], energy restriction and corticosterone treatment [2]. Measurements of 35S uptake in vivo also enabled the qualitative pattern of changes in proteoglycan synthesis in bone and muscle to be established. 2. Protein deficiency was examined by ad libitum feeding of 20%, 7%, 3.5% and 0.5% protein diets with measurements at 1, 3 and 7 days (all diets), and 14 and 21 days (0.5% protein). In bone this induced delayed inhibition of tibial growth with parallel inhibition of protein synthesis, as measured by the phenylalanine flooding dose method. This was mediated by reductions in both ribosomal capacity (RNA/protein ratio) and activity (protein synthesis/RNA) in the 0.5% protein group. The pattern of inhibition of proteoglycan sulphation, measured as 35S uptake 60 min after injection of a tracer dose of labelled sulphate, was similar to that of protein synthesis. 3. In muscle there was an intermediate graded inhibition of protein synthesis by protein deficiency, mediated by reductions in both ribosomal capacity and activity in the 0.5% protein group, which preceded growth inhibition in the 7% and 3.5% groups, and which was progressive with time. Transient increases in proteolysis contributed to the growth inhibition is some groups, but the rate fell eventually in the 0.5% group. The pattern of response of proteoglycan sulphation differed from protein synthesis with a delayed inhibition, but with subsequent marked reduction. 4. Energy restriction was induced by diets fed for 4 or 8 days at 75%, 50% and 25% ad libitum intakes with protein intakes held constant, and corticosterone treatment involved a dose of 10 mg day-1 100-1 g (subcutaneous) with ad libitum feeding. In bone this induced a pattern of length growth inhibition which was dissociated from inhibition of

  3. Comparison of sealing ability of bioactive bone cement, mineral trioxide aggregate and Super EBA as furcation repair materials: A dye extraction study

    Directory of Open Access Journals (Sweden)

    Janani Balachandran

    2013-01-01

    Settings and Design: In vitro, dye extraction study. Materials and Methods: Forty mandibular molars were randomly divided according to the material used to repair perforation: Group I-MTA, Group II-bioactive bone cement, Group III-Super EBA, Group IV-Control (furcation left unrepaired. All samples were subject to ortho grade and retrograde methylene blue dye challenge followed by dye extraction with 65% nitric acid. Samples were then analyzed using Ultra violet (UV Visible Spectrophotometer. Statistical Analysis Used: One way analysis of variance (ANOVA, Tukey-Kramer Multiple Comparisons Test. Results: MTA and bioactive bone cement showed almost similar and lower absorbance values in comparison to Super EBA. Conclusions: Bioactive bone cement provi ded an excellent seal for furcal perforation repair and at the same time it provided comfortable handling properties, which could overcome the potential disadvantages as faced with MTA.

  4. The effect of permanent grafting materials on the preservation of the buccal bone plate after tooth extraction: an experimental study in the dog.

    LENUS (Irish Health Repository)

    Bashara, Haitham

    2012-08-01

    The aim of the present study was to evaluate the effects of a novel bone substitute system (Natix(®)), consisting of porous titanium granules (PTG) and a bovine-derived xenograft (Bio-Oss(®)), on hard tissue remodelling following their placement into fresh extraction sockets in dogs.

  5. Towards plant protein refinery: Review on protein extraction using alkalo and potential enzymatic assistance

    NARCIS (Netherlands)

    Sari, Y.W.; Mulder, W.J.; Sanders, J.P.M.; Bruins, M.E.

    2015-01-01

    The globally increasing protein demands require additional resources to those currently available. Furthermore, the optimal usage of protein fractions from both traditional and new protein resources, such as algae and leaves, is essential. Here, we present an overview on alkaline plant protein extra

  6. Expression of bone morphogenic protein 2/4, transforming growth factor-β1, and bone matrix protein expression in healing area between vascular tibia grafts and irradiated bone-experimental model of osteonecrosis

    International Nuclear Information System (INIS)

    Purpose: For the surgical treatment of osteoradionecrosis after multimodal therapy of head-and-neck cancers, free vascular bone grafts are used to reconstruct osseous structures in the previously irradiated graft bed. Reduced, or even absent osseous healing in the transition area between the vascular graft and the irradiated graft bed represents a clinical problem. Inflammatory changes and fibrosis lead to delayed healing, triggered by bone morphogentic protein 2/4 (BMP2/4) and transforming growth factor (TGF)-β1. Given the well-known fibrosis-inducing activity of TGF-β1, an osteoinductive effect has been reported for BMP2/4. However, the influence of irradiation (RT) on this cytokine expression remains elusive. Therefore, the aim of the present in vivo study was to analyze the expression of BMP2/4, TGF-β1, collagen I, and osteocalcin in the transition area between the bone graft and the graft bed after RT. Methods and materials: Twenty Wistar rats (male, weight 300-500 g) were used in this study. A free vascular tibia graft was removed in all rats and maintained pedicled in the groin region. Ten rats underwent RT with 5 x 10 Gy to the right tibia, the remainder served as controls. After 4 weeks, the previously removed tibia grafts were regrafted into the irradiated (Group 1) and nonirradiated (Group 2) graft beds. The interval between RT and grafting was 4 weeks. After a 4-week osseous healing period, the bone grafts were removed, and the transition area between the nonirradiated graft and the irradiated osseous graft bed was examined histomorphometrically (National Institutes of Health imaging program) and immunohistochemically (avidin-biotin-peroxidase complex) for the expression of BMP2/4, TGF-β1, collagen I, and osteocalcin. Results: Absent or incomplete osseous healing of the graft was found in 9 of 10 rats after RT with 50 Gy and in 1 of 10 of the rats with nonirradiated osseous grafts. Histomorphometrically, the proportion of osseous healing in the

  7. Effectiveness and safety of recombinant human bone morphogenetic protein-2 for adults with lumbar spine pseudarthrosis following spinal fusion surgery

    Science.gov (United States)

    Balaji, V.; Kaila, R.; Wilson, L.

    2016-01-01

    Objectives We performed a systematic review of the literature to determine the safety and efficacy of bone morphogenetic protein (BMP) compared with bone graft when used specifically for revision spinal fusion surgery secondary to pseudarthrosis. Methods The MEDLINE, EMBASE and Cochrane Library databases were searched using defined search terms. The primary outcome measure was spinal fusion, assessed as success or failure in accordance with radiograph, MRI or CT scan review at 24-month follow-up. The secondary outcome measure was time to fusion. Results A total of six studies (three prospective and three retrospective) reporting on the use of BMP2 met the inclusion criteria (203 patients). Of these, four provided a comparison of BMP2 and bone graft whereas the other two solely investigated the use of BMP2. The primary outcome was seen in 92.3% (108/117) of patients following surgery with BMP2. Although none of the studies showed superiority of BMP2 to bone graft for fusion, its use was associated with a statistically quicker time to achieving fusion. BMP2 did not appear to increase the risk of complication. Conclusion The use of BMP2 is both safe and effective within the revision setting, ideally in cases where bone graft is unavailable or undesirable. Further research is required to define its optimum role. Cite this article: Mr P. Bodalia. Effectiveness and safety of recombinant human bone morphogenetic protein-2 for adults with lumbar spine pseudarthrosis following spinal fusion surgery: A systematic review. Bone Joint Res 2016;5:145–152. DOI: 10.1302/2046-3758.54.2000418. PMID:27121215

  8. Antiosteoporosis Effect of Radix Scutellariae Extract on Density and Microstructure of Long Bones in Tail-Suspended Sprague-Dawley Rats

    Directory of Open Access Journals (Sweden)

    Chen-Rui Li

    2013-01-01

    Full Text Available Radix Scutellariae (RS, a medicinal herb, is extensively employed in traditional Chinese medicines and modern herbal prescriptions. Two major flavonoids in RS were known to induce osteoblastic differentiation and inhibit osteoclast differentiation, respectively. This study aimed to investigate the effect of Radix Scutellariae extract (RSE against bone loss induced by mechanical inactivity or weightlessness. A hindlimb unloading tail-suspended rat model (TS was established to determine the effect of RSE on bone mineral density and bone microarchitecture. Treatment of RSE at 50 mg/kg/day and alendronate (ALE at 2 mg/kg/day as positive control for 42 days significantly increased the bone mineral density and mechanical strength compared with TS group. Enhanced bone turnover markers by TS treatment were attenuated by RSE and ALE administration. Deterioration of bone trabecula induced by TS was prevented. Moreover, both treatments counteracted the reduction of bone volume fraction, trabecular thickness and number, and connectivity density. In conclusion, RSE was demonstrated for the first time to prevent osteoporosis induced by TS treatment, which suggests the potential application of RSE in the treatment of disuse-induced osteoporosis.

  9. Automatic extraction of protein point mutations using a graph bigram association.

    Directory of Open Access Journals (Sweden)

    Lawrence C Lee

    2007-02-01

    Full Text Available Protein point mutations are an essential component of the evolutionary and experimental analysis of protein structure and function. While many manually curated databases attempt to index point mutations, most experimentally generated point mutations and the biological impacts of the changes are described in the peer-reviewed published literature. We describe an application, Mutation GraB (Graph Bigram, that identifies, extracts, and verifies point mutations from biomedical literature. The principal problem of point mutation extraction is to link the point mutation with its associated protein and organism of origin. Our algorithm uses a graph-based bigram traversal to identify these relevant associations and exploits the Swiss-Prot protein database to verify this information. The graph bigram method is different from other models for point mutation extraction in that it incorporates frequency and positional data of all terms in an article to drive the point mutation-protein association. Our method was tested on 589 articles describing point mutations from the G protein-coupled receptor (GPCR, tyrosine kinase, and ion channel protein families. We evaluated our graph bigram metric against a word-proximity metric for term association on datasets of full-text literature in these three different protein families. Our testing shows that the graph bigram metric achieves a higher F-measure for the GPCRs (0.79 versus 0.76, protein tyrosine kinases (0.72 versus 0.69, and ion channel transporters (0.76 versus 0.74. Importantly, in situations where more than one protein can be assigned to a point mutation and disambiguation is required, the graph bigram metric achieves a precision of 0.84 compared with the word distance metric precision of 0.73. We believe the graph bigram search metric to be a significant improvement over previous search metrics for point mutation extraction and to be applicable to text-mining application requiring the association of words.

  10. Imaging Bone Morphogenetic Protein 7 Induced Cell Cycle Arrest in Experimental Gliomas

    Directory of Open Access Journals (Sweden)

    Anke Klose

    2011-03-01

    Full Text Available Bone morphogenetic protein 7 (BMP-7 belongs to the superfamily of transforming growth factor β-like cytokines, which can act either as tumor suppressors or as tumor promoters depending on cell type and differentiation. Our investigations focused on analyzing the effects of BMP-7 during glioma cell proliferation in vitro and in vivo. BMP-7 treatment decreased the proliferation of Gli36ΔEGFR-LITG glioma cells up to 50%through a cell cycle arrest in the G1 phase but not by induction of apoptosis. This effect was mediated by the modulation of the expression and phosphorylation of cyclin-dependent kinase 2, cyclin-dependent kinase inhibitor p21, and downstream retinoblastoma protein. Furthermore, in vivo optical imaging of luciferase activity of Gli36ΔEGFR-LITG cells implanted intracranially into nude mice in the presence or absence of BMP-7 treatment corroborated the antiproliferative effects of this cytokine. This report clearly underlines the tumor-suppressive role of BMP-7 in glioma-derived cells. Taken together, our results indicate that manipulating the BMP/transforming growth factor β signaling cascade may serve as a new strategy for imaging-guided molecular-targeted therapy of malignant gliomas.

  11. Regulation of oligodendrocyte progenitor cell maturation by PPARδ: effects on bone morphogenetic proteins

    Directory of Open Access Journals (Sweden)

    Jill C Richardson

    2010-01-01

    Full Text Available In EAE (experimental autoimmune encephalomyelitis, agonists of PPARs (peroxisome proliferator-activated receptors provide clinical benefit and reduce damage. In contrast with PPARγ, agonists of PPARδ are more effective when given at later stages of EAE and increase myelin gene expression, suggesting effects on OL (oligodendrocyte maturation. In the present study we examined effects of the PPARδ agonist GW0742 on OPCs (OL progenitor cells, and tested whether the effects involve modulation of BMPs (bone morphogenetic proteins. We show that effects of GW0742 are mediated through PPARδ since no amelioration of EAE clinical scores was observed in PPARδ-null mice. In OPCs derived from E13 mice (where E is embryonic day, GW0742, but not the PPARγ agonist pioglitazone, increased the number of myelin-producing OLs. This was due to activation of PPARδ since process formation was reduced in PPARδ-null compared with wild-type OPCs. In both OPCs and enriched astrocyte cultures, GW0742 increased noggin protein expression; however, noggin mRNA was only increased in astrocytes. In contrast, GW0742 reduced BMP2 and BMP4 mRNA levels in OPCs, with lesser effects in astrocytes. These findings demonstrate that PPARδ plays a role in OPC maturation, mediated, in part, by regulation of BMP and BMP antagonists.

  12. Compound soft regenerated skull material for repairing dog skull defects using bone morphogenetic protein as an inductor and nanohydroxyapatite as a scaffold

    Institute of Scientific and Technical Information of China (English)

    Zhidong Shi; Mingwang Liu; Zhongzong Qin; Qinmei Wang; Ying Guo; Haiyong He; Zhonghe Yu

    2008-01-01

    BACKGROUND: In previous studies of skull defects and regeneration, bone morphogenetic protein as an inductor and nanohydroxyapatite as a scaffold have been cocultured with osteoblasts.OBJECTIVE: To verify the characteristics of the new skull regenerated material after compound soft regenerated skull material implantatiom.DESIGN, TIME AND SETTING: The self-control and inter-group control animal experiment was perfurmed at the Sun Yat-sen University, China from February to July 2007.MATERIALS: Twenty-tour healthy adult dogs of both genders weighing 15-20 kg were used in this study. Nanohydroxyapatite as a scaffold was cocultured with osteoblasts. Using demineralized canine bone matrix as a carrier, recombinant human bone morphogenetic protein-2 was employed to prepare compound soft regenerated skull material. Self-designed compound soft regenerated skull material was implanted in models of skull defects.METHODS: Animals were randomly assigned into two groups, Group A (n = 16) and Group B (n = 8).Bilateral 2.5-cm-diameter full-thickness parietal skull defects were made in all animals. In Group A, the right side was reconstructed with calcium alginate gel, osteoblasts, and nanomcter bone meal composite;the left side was reconstructed with calcium alginate gel, osteoblasts, nanometer bone meal and recombinant human bone morphogenetic protein-2 composite. In Group B, the right side was kept as a simple skull detect, and the left side was reconstructed with calcium alginate gel, osteoblasts, nanometer bone meal and recombinant human bone morphogenetic protein-2 composite.MAIN OUTCOME MEASURES: Bone regeneration and histopathological changes at the site of the skull defect were observed with an optical microscope and a scanning electron microscope after surgery.The ability to form bone was measured by alizarin red S staining. In vitro cultured osteoblasts were observed for morphology.RESULTS: One month following surgery, newly formed bone trabeculae mostly covered the

  13. A mineral-rich extract from the red marine algae Lithothamnion calcareum preserves bone structure and function in female mice on a Western-style diet.

    Science.gov (United States)

    Aslam, Muhammad Nadeem; Kreider, Jaclynn M; Paruchuri, Tejaswi; Bhagavathula, Narasimharao; DaSilva, Marissa; Zernicke, Ronald F; Goldstein, Steven A; Varani, James

    2010-04-01

    The purpose of this study was to determine whether a mineral-rich extract derived from the red marine algae Lithothamnion calcareum could be used as a dietary supplement for prevention of bone mineral loss. Sixty C57BL/6 mice were divided into three groups based on diet: the first group received a high-fat Western-style diet (HFWD), the second group was fed the same HFWD along with the mineral-rich extract included as a dietary supplement, and the third group was used as a control and was fed a low-fat rodent chow diet (AIN76A). Mice were maintained on the respective diets for 15 months. Then, long bones (femora and tibiae) from both males and females were analyzed by three-dimensional micro-computed tomography (micro-CT) and (bones from female mice) concomitantly assessed in bone strength studies. Tartrate-resistant acid phosphatase (TRAP), osteocalcin, and N-terminal peptide of type I procollagen (PINP) were assessed in plasma samples obtained from female mice at the time of sacrifice. To summarize, female mice on the HFWD had reduced bone mineralization and reduced bone strength relative to female mice on the low-fat chow diet. The bone defects in female mice on the HFWD were overcome in the presence of the mineral-rich supplement. In fact, female mice receiving the mineral-rich supplement in the HFWD had better bone structure/function than did female mice on the low-fat chow diet. Female mice on the mineral-supplemented HFWD had higher plasma levels of TRAP than mice of the other groups. There were no differences in the other two markers. Male mice showed little diet-specific differences by micro-CT. PMID:20180099

  14. A Mineral-Rich Extract from the Red Marine Algae Lithothamnion calcareum Preserves Bone Structure and Function in Female Mice on a Western-Style Diet

    Science.gov (United States)

    Aslam, Muhammad Nadeem; Kreider, Jaclynn M.; Paruchuri, Tejaswi; Bhagavathula, Narasimharao; DaSilva, Marissa; Zernicke, Ronald F.; Goldstein, Steven A.; Varani, James

    2010-01-01

    The purpose of this study was to determine whether a mineral-rich extract derived from the red marine algae Lithothamnion calcareum could be used as a dietary supplement for prevention of bone mineral loss. Sixty C57BL/6 mice were divided into three groups based on diet: the first group received a high-fat Western-style diet (HFWD), the second group was fed the same HFWD along with the mineral-rich extract included as a dietary supplement, and the third group was used as a control and was fed a low-fat rodent chow diet (AIN76A). Mice were maintained on the respective diets for 15 months. Then, long bones (femora and tibiae) from both males and females were analyzed by three-dimensional micro-computed tomography (micro-CT) and (bones from female mice) concomitantly assessed in bone strength studies. Tartrate-resistant acid phosphatase (TRAP), osteocalcin, and N-terminal peptide of type I procollagen (PINP) were assessed in plasma samples obtained from female mice at the time of sacrifice. To summarize, female mice on the HFWD had reduced bone mineralization and reduced bone strength relative to female mice on the low-fat chow diet. The bone defects in female mice on the HFWD were overcome in the presence of the mineral-rich supplement. In fact, female mice receiving the mineral-rich supplement in the HFWD had better bone structure/function than did female mice on the low-fat chow diet. Female mice on the mineral-supplemented HFWD had higher plasma levels of TRAP than mice of the other groups. There were no differences in the other two markers. Male mice showed little diet-specific differences by micro-CT. PMID:20180099

  15. Modification of an acetone-sodium dodecyl sulfate disruption method for cellular protein extraction from neuropathogenic Clostridium botulinum

    Science.gov (United States)

    An acetone-sodium dodecyl sulfate (SDS) disruption method was used for the extraction of cellular proteins from neurotoxigenic Clostridium botulinum. The amount of protein extracted per gram of dry weight and the protein profile as revealed by polyacrylamide gel electrophoresis (PAGE) was comparabl...

  16. How Does Alkali Aid Protein Extraction in Green Tea Leaf Residue: A Basis for Integrated Biorefinery of Leaves

    NARCIS (Netherlands)

    Zhang, C.; Sanders, J.P.M.; Xiao, T.T.; Bruins, M.E.

    2015-01-01

    Leaf protein can be obtained cost-efficiently by alkaline extraction, but overuse of chemicals and low quality of (denatured) protein limits its application. The research objective was to investigate how alkali aids protein extraction of green tea leaf residue, and use these results for further impr

  17. Mode of heparin attachment to nanocrystalline hydroxyapatite affects its interaction with bone morphogenetic protein-2.

    Science.gov (United States)

    Goonasekera, Chandhi S; Jack, Kevin S; Bhakta, Gajadhar; Rai, Bina; Luong-Van, Emma; Nurcombe, Victor; Cool, Simon M; Cooper-White, Justin J; Grøndahl, Lisbeth

    2015-01-01

    Heparin has a high affinity for bone morphogenetic protein-2 (BMP-2), which is a key growth factor in bone regeneration. The aim of this study was to investigate how the rate of release of BMP-2 was affected when adsorbed to nanosized hydroxyapatite (HAP) particles functionalized with heparin by different methods. Heparin was attached to the surface of HAP, either via adsorption or covalent coupling, via a 3-aminopropyltriethoxysilane (APTES) layer. The chemical composition of the particles was evaluated using X-ray photoelectron spectroscopy and elemental microanalysis, revealing that the heparin grafting densities achieved were dependent on the curing temperature used in the fabrication of APTES-modified HAP. Comparable amounts of heparin were attached via both covalent coupling and adsorption to the APTES-modified particles, but characterization of the particle surfaces by zeta potential and Brunauer-Emmett-Teller measurements indicated that the conformation of the heparin on the surface was dependent on the method of attachment, which in turn affected the stability of heparin on the surface. The release of BMP-2 from the particles after 7 days in phosphate-buffered saline found that 31% of the loaded BMP-2 was released from the APTES-modified particles with heparin covalently attached, compared to 16% from the APTES-modified particles with the heparin adsorbed. Moreover, when heparin was adsorbed onto pure HAP, it was found that the BMP-2 released after 7 days was 5% (similar to that from unmodified HAP). This illustrates that by altering the mode of attachment of heparin to HAP the release profile and total release of BMP-2 can be manipulated. Importantly, the BMP-2 released from all the heparin particle types was found by the SMAD 1/5/8 phosphorylation assay to be biologically active. PMID:26474791

  18. The expression and potential function of bone morphogenetic proteins 2 and 4 in bovine trophectoderm

    Directory of Open Access Journals (Sweden)

    Pennington Kathleen A

    2012-02-01

    Full Text Available Abstract Background Bone morphogenetic proteins (BMPs were first described for their roles in bone formation, but they now also are known to possess additional activities, including those relating to embryogenesis. The objectives of this work were to 1 determine if peri-attachment bovine conceptuses and bovine trophoblast cells (CT1 contain transcripts for BMP2 and 4, an innate inhibitor noggin (NOG, and BMP2/4 receptors (BMPRII, ACVR1, BMPR1A, BMPR1B, and 2 determine if BMP2 or 4 supplementation to CT1 cells affects cell proliferation, differentiation or trophoblast-specific gene expression. Methods RNA was isolated from day 17 bovine conceptuses and CT1 cells. After RT-PCR, amplified products were cloned and sequenced. In other studies CT1 cells were treated with BMP2 or 4 at various concentrations and effects on cell viability, cell differentiation and abundance of IFNT and CSH1 mRNA were evaluated. Results Transcripts for BMP2 and 4 were detected in bovine conceptuses and CT1 cells. Also, transcripts for each BMP receptor were detected in conceptuses and CT1 cells. Transcripts for NOG were detected in conceptuses but not CT1 cells. Cell proliferation was reduced by BMP4 but not BMP2 supplementation. Both factors reduced IFNT mRNA abundance but had no effect on CSH1 mRNA abundance in CT1 cells. Conclusions The BMP2/4 ligand and receptor system presides within bovine trophectoderm prior to uterine attachment. BMP4 negatively impacts CT1 cell growth and both BMPs affect IFNT mRNA abundance.

  19. Bone morphogenetic protein-2 gene controls tooth root development in coordination with formation of the periodontium

    Institute of Scientific and Technical Information of China (English)

    Audrey Rakian; Wu-Chen Yang; Jelica Gluhak-Heinrich; Yong Cui; Marie A Harris; Demitri Villarreal; Jerry Q Feng; Mary MacDougall; Stephen E Harris

    2013-01-01

    Formation of the periodontium begins following onset of tooth-root formation in a coordinated manner after birth. Dental follicle progenitor cells are thought to form the cementum, alveolar bone and Sharpey’s fibers of the periodontal ligament (PDL). However, little is known about the regulatory morphogens that control differentiation and function of these progenitor cells, as well as the progenitor cells involved in crown and root formation. We investigated the role of bone morphogenetic protein-2 (Bmp2) in these processes by the conditional removal of the Bmp2 gene using the Sp7-Cre-EGFP mouse model. Sp7-Cre-EGFP first becomes active at E18 in the first molar, with robust Cre activity at postnatal day 0 (P0), followed by Cre activity in the second molar, which occurs after P0. There is robust Cre activity in the periodontium and third molars by 2 weeks of age. When the Bmp2 gene is removed from Sp71 (Osterix1) cells, major defects are noted in root, cellular cementum and periodontium formation. First, there are major cell autonomous defects in root-odontoblast terminal differentiation. Second, there are major alterations in formation of the PDLs and cellular cementum, correlated with decreased nuclear factor IC (Nfic), periostin and a-SMA1 cells. Third, there is a failure to produce vascular endothelial growth factor A (VEGF-A) in the periodontium and the pulp leading to decreased formation of the microvascular and associated candidate stem cells in the Bmp2-cKOSp7-Cre-EGFP. Fourth, ameloblast function and enamel formation are indirectly altered in the Bmp2-cKOSp7-Cre-EGFP. These data demonstrate that the Bmp2 gene has complex roles in postnatal tooth development and periodontium formation.

  20. Basic science and spine literature document bone morphogenetic protein increases cancer risk

    Directory of Open Access Journals (Sweden)

    Nancy E Epstein

    2014-01-01

    Full Text Available Background: Increasingly, clinical articles document that bone morphogenetic protein (BMP/INFUSE: Medtronic, Memphis, TN, USA and its derivatives utilized in spinal surgery increase the risk of developing cancer. However, there is also a large body of basic science articles that also document that various types of BMP and other members of the TGF-Beta (transforming growth factor beta family promote the growth of different types of cancers. Methods: This review looks at many clinical articles citing BMP/INFUSE′s role, largely "off-label", in contributing to complications encountered during spinal surgery. Next, however, specific attention is given to the clinical and basic science literature regarding how BMP and its derivatives (e.g. members of the TGF-beta family may also impact the development of breast and other cancers. Results: Utilizing BMP/INFUSE in spine surgery increased the risk of cancers/new malignancy as documented in several studies. For example, Carragee et al. found that for single-level instrumented posterolateral fusions (PLF using high-dose rhBMP-2 (239 patients vs. autograft (control group; n = 224, the risks of new cancers at 2 and 5 years postoperatively were increased. In laboratory studies, BMP′s along with other members of the TGF-Beta family also modulated/contributed to the proliferation/differentiation of breast cancer (e.g. bone formation/turnover, breast cancer-related solid tumors, and metastases, lung, adrenal, and colon cancer. Conclusions: BMP/INFUSE when utilized clinically in spinal fusion surgery appears to promote cancer at higher rates than observed in the overall population. Furthermore, BMP and TGF-beta are correlated with increased cancer growth both in the clinic and the laboratory.

  1. A Protein Extract from Chicken Reduces Plasma Homocysteine in Rats

    Science.gov (United States)

    Lysne, Vegard; Bjørndal, Bodil; Vik, Rita; Nordrehaug, Jan Erik; Skorve, Jon; Nygård, Ottar; Berge, Rolf K.

    2015-01-01

    The present study aimed to evaluate effects of a water-soluble protein fraction of chicken (CP), with a low methionine/glycine ratio, on plasma homocysteine and metabolites related to homocysteine metabolism. Male Wistar rats were fed either a control diet with 20% w/w casein as the protein source, or an experimental diet where 6, 14 or 20% w/w of the casein was replaced with the same amount of CP for four weeks. Rats fed CP had reduced plasma total homocysteine level and markedly increased levels of the choline pathway metabolites betaine, dimethylglycine, sarcosine, glycine and serine, as well as the transsulfuration pathway metabolites cystathionine and cysteine. Hepatic mRNA level of enzymes involved in homocysteine remethylation, methionine synthase and betaine-homocysteine S-methyltransferase, were unchanged, whereas cystathionine gamma-lyase of the transsulfuration pathway was increased in the CP treated rats. Plasma concentrations of vitamin B2, folate, cobalamin, and the B-6 catabolite pyridoxic acid were increased in the 20% CP-treated rats. In conclusion, the CP diet was associated with lower plasma homocysteine concentration and higher levels of serine, choline oxidation and transsulfuration metabolites compared to a casein diet. The status of related B-vitamins was also affected by CP. PMID:26053618

  2. A Protein Extract from Chicken Reduces Plasma Homocysteine in Rats

    Directory of Open Access Journals (Sweden)

    Vegard Lysne

    2015-06-01

    Full Text Available The present study aimed to evaluate effects of a water-soluble protein fraction of chicken (CP, with a low methionine/glycine ratio, on plasma homocysteine and metabolites related to homocysteine metabolism. Male Wistar rats were fed either a control diet with 20% w/w casein as the protein source, or an experimental diet where 6, 14 or 20% w/w of the casein was replaced with the same amount of CP for four weeks. Rats fed CP had reduced plasma total homocysteine level and markedly increased levels of the choline pathway metabolites betaine, dimethylglycine, sarcosine, glycine and serine, as well as the transsulfuration pathway metabolites cystathionine and cysteine. Hepatic mRNA level of enzymes involved in homocysteine remethylation, methionine synthase and betaine-homocysteine S-methyltransferase, were unchanged, whereas cystathionine gamma-lyase of the transsulfuration pathway was increased in the CP treated rats. Plasma concentrations of vitamin B2, folate, cobalamin, and the B-6 catabolite pyridoxic acid were increased in the 20% CP-treated rats. In conclusion, the CP diet was associated with lower plasma homocysteine concentration and higher levels of serine, choline oxidation and transsulfuration metabolites compared to a casein diet. The status of related B-vitamins was also affected by CP.

  3. Modulation of function of multidrug resistance associated-proteins by Kaempferia parviflora extracts and their components

    OpenAIRE

    Patanasethanont, Denpong; Nagai, Junya; Matsuura, Chie; Fukui, Kyoko; Sutthanut, Khaetthareeya; Sripanidkulchai, Bung-Orn; Yumoto, Ryoko; Takano, Mikihisa

    2007-01-01

    In this study, the effects of extracts and flavone derivatives from the rhizome of Kaempferia parviflora on multidrug resistance associated-proteins (MRP)-mediated transport in A549 cells were examined. The cells employed express MRP1 and MRP2, but not P-glycoprotein. The cellular accumulation of calcein, an MRP substrate, was significantly increased by various MRP inhibitors without being affected by verapamil, a typical P-glycoprotein inhibitor. Ethanol and aqueous extracts from Kaempferia ...

  4. A Rapid Screening Assay to Search for Phosphorylated Proteins in Tissue Extracts

    OpenAIRE

    Garaguso, Ignazio; Borlak, Juergen

    2012-01-01

    Reversible protein phosphorylation is an essential mechanism in the regulation of diverse biological processes, nonetheless is frequently altered in disease. As most phosphoproteome studies are based on optimized in-vitro cell culture studies new methods are in need to improve de novo identification and characterization of phosphoproteins in extracts from tissues. Here, we describe a rapid and reliable method for the detection of phosphoproteins in tissue extract based on an experimental stra...

  5. Effect of green tea or rosemary extract on protein oxidation in Bologna type sausages prepared from oxidatively stressed pork.

    Science.gov (United States)

    Jongberg, Sisse; Tørngren, Mari Ann; Gunvig, Annemarie; Skibsted, Leif H; Lund, Marianne N

    2013-03-01

    Bologna type sausages were prepared from oxidatively stressed pork (UV-irradiation, 48 h, 5 °C) using a traditional recipe (control) or the same recipe but added green tea extract (500 ppm total phenolic compounds) or rosemary extract (400 ppm total phenolic compounds). Green tea and rosemary extracts protected against formation of TBARS and protein carbonyls. On the contrary, increased thiol loss and a distinct loss of myosin heavy chain and actin due to polymerization by reducible bonds as determined by SDS-page were found by addition of green tea extract. The enhanced protein polymerization was ascribed to the reaction between quinone compounds from the plant extracts and protein thiol groups to yield phenol-mediated protein polymerization. Analysis by ESR spectroscopy revealed increased radical intensities in sausages added plant extracts, which was ascribed to originate from protein-bound phenoxyl radicals, which may protect against other oxidatively induced protein modifications. PMID:23273462

  6. Proteomic Analysis of Terminalia chebula Extract-Dependent Changes in Human Lymphoblastic T Cell Protein Expression

    Science.gov (United States)

    Das, Nando Dulal; Jung, Kyoung Hwa; Park, Ji Hyun; Choi, Mi Ran; Lee, Hyung Tae; Kim, Moo Sung; Lee, Sang Rin

    2012-01-01

    Abstract Terminalia chebula is a native plant from southern Asia to southwestern China that is used in traditional medicine for the treatment of malignant tumors and diabetes. This plant also has antibacterial and immunomodulatory properties. The present study assessed T. chebula extract-dependent protein expression changes in Jurkat cells. Matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry and Ingenuity Pathways Analysis (IPA) were performed to assess protein expression and networks, respectively. A comparative proteomic profile was determined in T. chebula extract (50 μg/mL)-treated and control cells; the expressions of β-tubulin, ring finger and CHY zinc finger domain containing 1, and insulin-like growth factor 1 receptor kinase were significantly down-regulated in T. chebula extract-treated Jurkat cells. Moreover, the molecular basis for the T. chebula extract-dependent protein expression changes in Jurkat cells was determined by IPA. Treatment with the T. chebula extract significantly inhibited nuclear factor-κB activity and affected the proteomic profile of Jurkat cells. The molecular network signatures and functional proteomics obtained in this study may facilitate the evaluation of potential antitumor therapeutic targets and elucidate the molecular mechanism of T. chebula extract-dependent effects in Jurkat cells. PMID:22471968

  7. Extraction of Protein from Common Plant Leaves for Use as Human Food

    Directory of Open Access Journals (Sweden)

    Abdel E. Ghaly

    2010-01-01

    Full Text Available Problem statement: Protein deficiency is one of the major nutritional problems in the developing world. The most disastrous consequences occur in children where protein malnutrition manifests itself in forms of two notorious diseases: Marasmus and kwashiorkor. Expansion of present agricultural practices into marginal lands is expected to solve this chronic protein shortage. The process of photosynthesis is the only non depletable protein source and can supply some essential amino acids as well as provide adequate nitrogen in the diet for synthesis of non essential amino acids in addition to vitamins and minerals. Approach: The aim of this study was to assess the nutritional values of common plant leaves and determine the feasibility of using them as a protein supplement. Six plants were cultivated and tested for their nutritional quality: amaranth, cowpea, sugar been, pumpkin, sweet potato and cabbage. Results: Pumpkin leaves recorded the highest protein yield (11.75% followed by amaranth (10.5% The protein contents in the leaves of sweet potato, cowpea, cabbage and sugar been were much lower (7.85, 6.95, 5.60 and 3.45%, respectively. Conclusion: Except for sugar bean, all plants had higher extractable protein content than cabbage. Proper use of the growing season can achieve high protein yield. Clean cutting will ensure that the leaves do not deteriorate within a few hours when kept in cool place and freezing leaves can improve protein extractability. The test plants should be given a trial as protein sources for human. Plant leaves have vitamins, minerals and essential amino acids and when consumed in adequate amounts they can supplement protein especially in areas where the environment is very hostile to livestock keeping or where fish protein is lacking. The optimum harvest age, fertilizer requirements and the possibility of combining two or more plants together to improve protein content should be studied. The amino acid, minerals and

  8. Application of thiopropyl sepharose 6B for removal of PCR inhibitors from DNA extracts of a thigh bone recovered from the sea

    DEFF Research Database (Denmark)

    Sørensen, Erik; Hansen, Steen Holger; Eriksen, Birthe;

    2003-01-01

    PCR amplification of DNA from forensic samples often proves difficult due to the presence of inhibitors of the polymerase chain reaction. One possible way to remove PCR inhibitors from a DNA extract is the use of the affinity resin thiopropyl sepharose 6B (TS), which has been used previously for...... the removal of PCR inhibitors in DNA extracts originating from stains on clothing. Here we show that TS is efficient also for the removal of inhibitors from PCR extracts from a highly decomposed human thigh bone. TS treatment, however, leads to a substantial loss of DNA making the technique best...... suited when substantial amounts of DNA are present....

  9. A review on rice bran protein: its properties and extraction methods.

    Science.gov (United States)

    Fabian, Cynthia; Ju, Yi-Hsu

    2011-01-01

    Rice bran protein has been found to be of high quality and of importance for food and pharmaceutical applications. It is a plant protein that can be derived from rice bran, an abundant and cheap agricultural byproduct. The protein content in rice bran is about 10-15% and it consists of 37% water-soluble, 31% salt-soluble, 2% alcohol-soluble, and 27% alkali-soluble storage proteins. Its unique property as being hypoallergenic and having anti-cancer activity makes it a superior cereal protein that may find a wide range of applications. There were already reports on the extraction of rice bran protein several decades ago. However, as of now, commercial rice bran protein is still unavailable in the market. This review is aimed at providing valuable discussions on rice bran protein, that is, storage protein, its various properties, and extraction methods for the development of an effective processing scheme. Also, an update on the current processing methods is also included. PMID:21888532

  10. Rice proteins, extracted by alkali and α-amylase, differently affect in vitro antioxidant activity.

    Science.gov (United States)

    Wang, Zhengxuan; Liu, Ye; Li, Hui; Yang, Lin

    2016-09-01

    Alkali treatment and α-amylase degradation are different processes for rice protein (RP) isolation. The major aim of this study was to determine the influence of two different extraction methods on the antioxidant capacities of RPA, extracted by alkaline (0.2% NaOH), and RPE, extracted by α-amylase, during in vitro digestion for 2h with pepsin and for 3h with pancreatin. Upon pepsin-pancreatin digestion, the protein hydrolysates (RPA-S, RPE-S), which were the supernatants in the absence of undigested residue, and the whole protein digests (RPA, RPE), in which undigested residue remained, were measured. RPE exhibited the stronger antioxidant responses to free radical scavenging activity, metal chelating activity, and reducing power, whereas the weakest antioxidant capacities were produced by RPE-S. In contrast, no significant differences in antioxidant activity were observed between RPA and RPA-S. The present study demonstrated that the in vitro antioxidant responses induced by the hydrolysates and the protein digests of RPs could be affected differently by alkali treatment and α-amylase degradation, suggesting that the extraction is a vital processing step to modify the antioxidant capacities of RPs. The results of the current study indicated that the protein digests, in which undigested residues remained, could exhibit more efficacious antioxidant activity compared to the hydrolysates. PMID:27041309

  11. Effect of pomegranate peel extract on lipid and protein oxidation in beef meatballs during refrigerated storage.

    Science.gov (United States)

    Turgut, Sebahattin Serhat; Soyer, Ayla; Işıkçı, Fatma

    2016-06-01

    Antioxidant effect of pomegranate peel extract (PE) to retard lipid and protein oxidation was investigated in meatballs during refrigerated storage at 4±1°C. Concentrated lyophilised water extract of pomegranate peel was incorporated into freshly minced beef meat at 0.5% and 1% concentrations and compared with 0.01% butylated hydroxytoluene (BHT) as a reference and control (without any antioxidant). PE showed high phenolic content and antioxidant activity. In PE added samples, thiobarbituric acid reactive substances (TBARS) value, peroxide formation, loss of sulfhydryl groups and formation of protein carbonyls were lower than control (PSensory evaluation with respect to colour and rancid odour revealed that PE incorporation in meatballs prolonged the refrigerated storage up to 8 days. Addition of both 0.5 and 1% PE in meatballs reduced lipid and protein oxidation and improved sensory scores. These results indicated that PE was effective on retarding lipid and protein oxidation. PMID:26878610

  12. Extraction and comparison of proteins from natural rubber latex by conventional and ionizing radiation methods

    International Nuclear Information System (INIS)

    Several proteins in natural rubber latex (NRL) have been assigned to be significant allergens. It is known that proteins submitted to ionizing radiation suffer denaturation and immunochemical modification resulting in low antigenic reactivity. The aim of this study was to extract and compare water extractable proteins from NRL films vulcanized by conventional and by ionizing radiation methods. SDS-polyacrylamide gel electrophoresis (SDS--PAGE) and high pressure liquid chromatography (HPLC) showed a diffuse protein band of about 14 KDa, which we believe is rubber elongation factor (REF), in both eluates, but smaller in latex film vulcanized by ionizing radiation. REF has been suggested to be a major latex allergen. These data suggest that ionizing radiation vulcanization could be an useful method for the production of NRL goods with low antigenicity. (author). 8 refs., 2 figs., 1 tab

  13. In vitro and in vivo evaluation of bone formation using solid freeform fabrication-based bone morphogenic protein-2 releasing PCL/PLGA scaffolds

    International Nuclear Information System (INIS)

    The aim of this study was to develop novel polycaprolactone/poly(lactic-co-glycolic acid) (PCL/PLGA) scaffolds with a heparin–dopamine (Hep–DOPA) conjugate for controlled release of bone morphogenic protein-2 (BMP-2) to enhance osteoblast activity in vitro and also bone formation in vivo. PCL/PLGA scaffolds were prepared by a solid freeform fabrication method. The PCL/PLGA scaffolds were functionalized with Hep–DOPA and then BMP-2 was sequentially coated onto the Hep–DOPA/PCL/PLGA scaffolds. The characterization and surface elemental composition of all scaffolds were evaluated by scanning electron microscope and x-ray photoelectron spectroscopy. The osteoblast activities on all scaffolds were assessed by cell proliferation, alkaline phosphatase (ALP) activity and calcium deposition in vitro. To demonstrate bone formation in vivo, plain radiograph, micro-computed tomography (micro-CT) evaluation and histological studies were performed after the implantation of all scaffolds on a rat femur defect. Hep–DOPA/PCL/PLGA had more controlled release of BMP-2, which was quantified by enzyme-linked immunosorbent assay, compared with Hep/PCL/PLGA. The in vitro results showed that osteoblast-like cells (MG-63 cells) grown on BMP-2/Hep–DOPA/PCL/PLGA had significantly enhanced ALP activity and calcium deposition compared with those on BMP-2/Hep/PCL/PLGA and PCL/PLGA. In addition, the plain radiograph, micro-CT evaluation and histological studies demonstrated that the implanted BMP-2/Hep–DOPA/PCL/PLGA on rat femur had more bone formation than BMP-2/Hep/PCL/PLGA and PCL/PLGA in vivo. (paper)

  14. Optimization for Ultrasound-assisted Calcium Hydroxide Extraction of Protein from Shrimp Waste using Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Xiaopeng Cui

    2014-02-01

    Full Text Available Each year a considerable number of shrimp wastes were discarded. It caused a waste of resources, but also led to environmental pollution. In order to make a reasonable use of shrimp waste, protein was extracted from Vannamei waste by calcium hydroxide and ultrasonic-assisted, then used response surface methodology to optimize experiments to find the optimum extraction method. The optimal condition, predicted protein extraction rate of 37%, was obtained under the optimum conditions of the extraction time of 81 min, the extraction temperature of 60.00°C and the extraction concentration of 0.18 g/g dry matter.

  15. Proteomic Analysis of Terminalia chebula Extract-Dependent Changes in Human Lymphoblastic T Cell Protein Expression

    OpenAIRE

    Das, Nando Dulal; Jung, Kyoung Hwa; Park, Ji Hyun; Choi, Mi Ran; LEE, HYUNG TAE; Kim, Moo Sung; Lee, Sang Rin; Chai, Young Gyu

    2012-01-01

    Terminalia chebula is a native plant from southern Asia to southwestern China that is used in traditional medicine for the treatment of malignant tumors and diabetes. This plant also has antibacterial and immunomodulatory properties. The present study assessed T. chebula extract-dependent protein expression changes in Jurkat cells. Matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry and Ingenuity Pathways Analysis (IPA) were performed to assess protein expression and ...

  16. The Ginkgo biloba Extract EGb 761 Modulates Proteasome Activity and Polyglutamine Protein Aggregation

    OpenAIRE

    Marcel Stark; Christian Behl

    2014-01-01

    The standardized Ginkgo biloba extract EGb 761 has well-described antioxidative activities and effects on different cytoprotective signaling pathways. Consequently, a potential use of EGb 761 in neurodegenerative diseases has been proposed. A common characteristic feature of a variety of such disorders is the pathologic formation of protein aggregates, suggesting a crucial role for protein homeostasis. In this study, we show that EGb 761 increased the catalytic activity of the proteasome and ...

  17. A new concept for implant fixation: bone-to-bone biologic fixation.

    Science.gov (United States)

    Kim, D-Y; Kim, J-R; Jang, K Y; Lee, K-B

    2015-01-01

    Many attempts have been made to reduce complications of bone implant, such as pedicle screw loosening. To address this problem, the authors suggest a new concept of bone-to-bone biologic fixation using recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded cannulated pedicle screws. Recombinant human bone morphogenetic protein-2 is an osteoinductive cytokine. Four types of titanium pedicle screws were tested (uncannulated, cannulated with no loading, beta-tricalcium phosphate (TCP)-loaded, and TCP/BMP2 loaded) using 16 miniature pigs. Radiological evaluation was conducted to assess the fusion and loosening of pedicle screws. Twelve weeks after implantation, peak torsional extraction torque was measured, and the pedicle screw and bone interface was evaluated by micro-computed tomography (µCT) and histologic examination. The mean value of the radiological score was significantly greater in the TCP/BMP2 loaded group at 12 weeks post-operation compared to those in the other groups. CT images showed distinct bone formation surrounding TCP/BMP2 loaded cannulated pedicle screws compared to the other groups. Mean extraction torsional peak torque at 12 weeks postoperative was more than 10-fold higher in the TCP/BMP2 loaded pedicle screw group than in the other groups. Bone surface and bone volume, as quantitated through µCT, were higher in the TCP/BMP2 loaded group. Histologic examination revealed bone-to-bone fixation at the interface of pedicle screws and pre-existing bone. Bone-to-bone biologic fixation through the holes of TCP/BMP2 loaded pedicle screws significantly increased fixation strength and represents a novel method that can be applied to osteoporotic or tumour spine surgeries. PMID:25978116

  18. A new concept for implant fixation: bone-to-bone biologic fixation

    Directory of Open Access Journals (Sweden)

    D-Y Kim

    2015-05-01

    Full Text Available Many attempts have been made to reduce complications of bone implant, such as pedicle screw loosening. To address this problem, the authors suggest a new concept of bone-to-bone biologic fixation using recombinant human bone morphogenetic protein-2 (rhBMP-2-loaded cannulated pedicle screws. Recombinant human bone morphogenetic protein-2 is an osteoinductive cytokine. Four types of titanium pedicle screws were tested (uncannulated, cannulated with no loading, beta-tricalcium phosphate (TCP-loaded, and TCP/BMP2 loaded using 16 miniature pigs. Radiological evaluation was conducted to assess the fusion and loosening of pedicle screws. Twelve weeks after implantation, peak torsional extraction torque was measured, and the pedicle screw and bone interface was evaluated by micro-computed tomography (µCT and histologic examination. The mean value of the radiological score was significantly greater in the TCP/BMP2 loaded group at 12 weeks post-operation compared to those in the other groups. CT images showed distinct bone formation surrounding TCP/BMP2 loaded cannulated pedicle screws compared to the other groups. Mean extraction torsional peak torque at 12 weeks postoperative was more than 10-fold higher in the TCP/BMP2 loaded pedicle screw group than in the other groups. Bone surface and bone volume, as quantitated through µCT, were higher in the TCP/BMP2 loaded group. Histologic examination revealed bone-to-bone fixation at the interface of pedicle screws and pre-existing bone. Bone-to-bone biologic fixation through the holes of TCP/BMP2 loaded pedicle screws significantly increased fixation strength and represents a novel method that can be applied to osteoporotic or tumour spine surgeries.

  19. Immunization with FSHβ fusion protein antigen prevents bone loss in a rat ovariectomy-induced osteoporosis model

    International Nuclear Information System (INIS)

    Highlights: •A GST-FSH fusion protein was successfully expressed in E. coli. •Immunization with GST-FSH antigen can raise high-titer anti-FSH polyclonal sera. •Anti-FSH polyclonal sera can neutralize osteoclastogenic effect of FSH in vitro. •FSH immunization can prevent bone loss in a rat osteoporosis model. -- Abstract: Osteoporosis, a metabolic bone disease, threatens postmenopausal women globally. Hormone replacement therapy (HTR), especially estrogen replacement therapy (ERT), is used widely in the clinic because it has been generally accepted that postmenopausal osteoporosis is caused by estrogen deficiency. However, hypogonadal α and β estrogen receptor null mice were only mildly osteopenic, and mice with either receptor deleted had normal bone mass, indicating that estrogen may not be the only mediator that induces osteoporosis. Recently, follicle-stimulating hormone (FSH), the serum concentration of which increases from the very beginning of menopause, has been found to play a key role in postmenopausal osteoporosis by promoting osteoclastogenesis. In this article, we confirmed that exogenous FSH can enhance osteoclast differentiation in vitro and that this effect can be neutralized by either an anti-FSH monoclonal antibody or anti-FSH polyclonal sera raised by immunizing animals with a recombinant GST-FSHβ fusion protein antigen. Moreover, immunizing ovariectomized rats with the GST-FSHβ antigen does significantly prevent trabecular bone loss and thereby enhance the bone strength, indicating that a FSH-based vaccine may be a promising therapeutic strategy to slow down bone loss in postmenopausal women

  20. Immunization with FSHβ fusion protein antigen prevents bone loss in a rat ovariectomy-induced osteoporosis model

    Energy Technology Data Exchange (ETDEWEB)

    Geng, Wenxin; Yan, Xingrong; Du, Huicong; Cui, Jihong; Li, Liwen, E-mail: liven@nwu.edu.cn; Chen, Fulin, E-mail: chenfl@nwu.edu.cn

    2013-05-03

    Highlights: •A GST-FSH fusion protein was successfully expressed in E. coli. •Immunization with GST-FSH antigen can raise high-titer anti-FSH polyclonal sera. •Anti-FSH polyclonal sera can neutralize osteoclastogenic effect of FSH in vitro. •FSH immunization can prevent bone loss in a rat osteoporosis model. -- Abstract: Osteoporosis, a metabolic bone disease, threatens postmenopausal women globally. Hormone replacement therapy (HTR), especially estrogen replacement therapy (ERT), is used widely in the clinic because it has been generally accepted that postmenopausal osteoporosis is caused by estrogen deficiency. However, hypogonadal α and β estrogen receptor null mice were only mildly osteopenic, and mice with either receptor deleted had normal bone mass, indicating that estrogen may not be the only mediator that induces osteoporosis. Recently, follicle-stimulating hormone (FSH), the serum concentration of which increases from the very beginning of menopause, has been found to play a key role in postmenopausal osteoporosis by promoting osteoclastogenesis. In this article, we confirmed that exogenous FSH can enhance osteoclast differentiation in vitro and that this effect can be neutralized by either an anti-FSH monoclonal antibody or anti-FSH polyclonal sera raised by immunizing animals with a recombinant GST-FSHβ fusion protein antigen. Moreover, immunizing ovariectomized rats with the GST-FSHβ antigen does significantly prevent trabecular bone loss and thereby enhance the bone strength, indicating that a FSH-based vaccine may be a promising therapeutic strategy to slow down bone loss in postmenopausal women.

  1. Soy protein is beneficial but high-fat diet and voluntary running are detrimental to bone structure in mice.

    Science.gov (United States)

    Yan, Lin; Graef, George L; Nielsen, Forrest H; Johnson, LuAnn K; Cao, Jay

    2015-06-01

    Physical activity and soy protein isolate (SPI) augmentation have been reported to be beneficial for bone health. We hypothesized that combining voluntary running and SPI intake would alleviate detrimental changes in bone induced by a high-fat diet. A 2 × 2 × 2 experiment was designed with diets containing 16% or 45% of energy as corn oil and 20% SPI or casein fed to sedentary or running male C57BL/6 mice for 14 weeks. Distal femurs were assessed for microstructural changes. The high-fat diet significantly decreased trabecular number (Tb.N) and bone mineral density (BMD) and increased trabecular separation (Tb.Sp). Soy protein instead of casein, regardless of fat content, in the diet significantly increased bone volume fraction, Tb.N, connectivity density, and BMD and decreased Tb.Sp. Voluntary running, regardless of fat content, significantly decreased bone volume fraction, Tb.N, connectivity density, and BMD and increased Tb.Sp. The high-fat diet significantly decreased osteocalcin and increased tartrate-resistant acid phosphatase 5b (TRAP 5b) concentrations in plasma. Plasma concentrations of osteocalcin were increased by both SPI and running. Running alleviated the increase in TRAP 5b induced by the high-fat diet. These findings demonstrate that a high-fat diet is deleterious, and SPI is beneficial to trabecular bone properties. The deleterious effect of voluntary running on trabecular structural characteristics indicates that there may be a maximal threshold of running beyond which beneficial effects cease and detrimental effects occur. Increases in plasma osteocalcin and decreases in plasma TRAP 5b in running mice suggest that a compensatory response occurs to counteract the detrimental effects of excessive running. PMID:25957968

  2. Petroleum ether extract of Cissus quadrangularis (LINN stimulates the growth of fetal bone during intra uterine developmental period: a morphometric analysis

    Directory of Open Access Journals (Sweden)

    Bhagath Kumar Potu

    2008-01-01

    Full Text Available OBJECTIVE: The aim of the present study was to analyze the effect Cissus quadrangularis plant petroleum ether extract on the development of long bones during the intra-uterine developmental stage in rats. METHODS: Pregnant rats (n=12 were randomly assigned into either a control group (n=6 or a Cissus quadrangularis treatment (n=6 group. Pregnant rats in the Cissus quadrangularis group were treated with Cissus quadrangularis petroleum ether extract at a dose of 500 mg/kg body weight from gestation day 9 until delivery. The animals in the control group received an equal volume of saline. Newborn pups were collected from both groups for alizarin red S - alcian blue staining to differentiate ossified and unossified cartilage. The ossified cartilage (bone was morphometrically analyzed using Scion image software. RESULTS: Morphometric analysis revealed that the percentage of the total length of ossified cartilage (bone in pups born to treated dams was significantly higher (P<0.001- -0.0001 than that of the control group. CONCLUSION: The results of the present study suggest that maternal administration of Cissus quadrangularis petroleum ether extract during pregnancy can stimulate the development of fetal bone growth during the intra-uterine developmental period.

  3. Expression of genes for bone morphogenetic proteins BMP-2, BMP-4 and BMP-6 in various parts of the human skeleton

    Directory of Open Access Journals (Sweden)

    Włodarski Krzysztof

    2007-12-01

    Full Text Available Abstract Background Differences in duration of bone healing in various parts of the human skeleton are common experience for orthopaedic surgeons. The reason for these differences is not obvious and not clear. Methods In this paper we decided to measure by the use of real-time RT-PCR technique the level of expression of genes for some isoforms of bone morphogenetic proteins (BMPs, whose role is proven in bone formation, bone induction and bone turnover. Seven bone samples recovered from various parts of skeletons from six cadavers of young healthy men who died in traffic accidents were collected. Activity of genes for BMP-2, -4 and -6 was measured by the use of fluorescent SYBR Green I. Results It was found that expression of m-RNA for BMP-2 and BMP-4 is higher in trabecular bone in epiphyses of long bones, cranial flat bones and corpus mandibulae then in the compact bone of diaphyses of long bones. In all samples examined the expression of m-RNA for BMP-4 was higher than for BMP-2. Conclusion It was shown that m-RNA for BMP-6 is not expressed in the collected samples at all. It is postulated that differences in the level of activation of genes for BMPs is one of the important factors which determine the differences in duration of bone healing of various parts of the human skeleton.

  4. Efficient extraction of proteins from recalcitrant plant tissue for subsequent analysis by two-dimensional gel electrophoresis.

    Science.gov (United States)

    Parkhey, Suruchi; Chandrakar, Vibhuti; Naithani, S C; Keshavkant, S

    2015-10-01

    Protein extraction for two-dimensional electrophoresis from tissues of recalcitrant species is quite problematic and challenging due to the low protein content and high abundance of contaminants. Proteomics in Shorea robusta is scarcely conducted due to the lack of a suitable protein preparation procedure. To establish an effective protein extraction protocol suitable for two-dimensional electrophoresis in Shorea robusta, four procedures (borate buffer/trichloroacetic acid extraction, organic solvent/trichloroacetic acid precipitation, sucrose/Tris/phenol, and organic solvent/phenol/sodium dodecyl sulfate) were evaluated. Following these, proteins were isolated from mature leaves and were analyzed for proteomics, and also for potential contaminants, widely reported to hinder proteomics. The borate buffer/trichloroacetic acid extraction had the lowest protein yield and did not result in any banding even in one-dimensional electrophoresis. In contrast, organic solvent/phenol/sodium dodecyl sulfate extraction allowed the highest protein yield. Moreover, during proteomics, organic solvent/phenol/sodium dodecyl sulfate extracted protein resolved the maximum number (144) of spots. Further, when proteins were evaluated for contaminants, significant (77-95%) reductions in the nucleic acids, phenol, and sugars were discernible with refinement in extraction procedure. Accumulated data suggested that the organic solvent/phenol/sodium dodecyl sulfate extraction was the most effective protocol for protein isolation for proteomics of Shorea robusta and can be used for plants that have a similar set of contaminants. PMID:26257211

  5. The extraction of different proteins in selenium enriched peanuts and their antioxidant properties.

    Science.gov (United States)

    Zhao, P; Wang, Y; Zhang, Y; Guo, T; Zhang, Z; Zhang, W J; Zhang, X G; Ashraf, Muhammad Aqeel

    2016-05-01

    In this study, the selenium enriched peanuts and the different solubility proteins extracted from them were investigated. The dried defatted selenium enriched peanuts (SeP) powder (0.3147 μg/g) had a 2.5-fold higher mean total selenium concentration than general peanuts (GP) power (0.1233 μg/g). The SeP had higher concentration of selenium, manganese and zinc than that of GP, but less calcium. The rate of extraction of protein was 23.39% for peanuts and alkali soluble protein was the main component of protein in SeP, which accounted for 92.82% of total soluble protein and combined selenium was 77.33% of total selenium protein. In different forms of proteins from SeP, the WSePr due to higher concentration of selenium had higher DPPH free-radical scavenging activity, higher reducing activity and longer induction time than other proteins. PMID:27081360

  6. Kartogenin, transforming growth factor-β1 and bone morphogenetic protein-7 coordinately enhance lubricin accumulation in bone-derived mesenchymal stem cells.

    Science.gov (United States)

    Liu, Chun; Ma, Xueqin; Li, Tao; Zhang, Qiqing

    2015-09-01

    Osteoarthritis, a common joint degeneration, can cause breakdown of articular cartilage with the presence of lubricin metabolic abnormalities. Lubricin is a multi-level chondroprotective mucinous glycoprotein in articular joints. Joint defect and infection is elevated and accompanied by accelerated cartilage lesions involving degradation and loss of lubricin. However, a novel, heterocyclic compound called kartogenin (KGN) was discovered to stimulate chondrogenic differentiation of bone-derived mesenchymal stem cells (BMSCs). And the synergistic effect of transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7) could provoke lubricin accumulation. This paper attempted to explore the connection between accumulation of lubricin and the effect of TGF-β1, BMP-7 and/or KGN. Hence, we investigated the expression and secretion of lubricin in BMSCs treated with different combinations of TGF-β1, BMP-7, and/or KGN. Using an in vitro BMSCs system, we observed the content of lubricin from BMSCs treated with TGF-β1, BMP-7, and KGN was the highest at both the protein level and the gene level. The accumulation of lubricin was enhanced coordinately by the increase of synthesis and decrease of degradation possibly via c-Myc and adamts5 pathway. These results further suggested that supplementation of the defect parts with lubricin by using growth factors and small molecules showed a promising potential on preventing joint deterioration in patients with acquired or genetic deficiency of lubricin in the future of regenerative medicine. PMID:25857705

  7. Ridge preservation with acellular dermal matrix and anorganic bone matrix cell-binding peptide P-15 after tooth extraction in humans. A histologic and morphometric study

    Directory of Open Access Journals (Sweden)

    Arthur B. Novaes Jr

    2012-06-01

    Full Text Available Aim: The aim of this study was to analyze by histomorphometric parameters the use of acellular dermal matrix (ADM with or without anorganic bovine bone matrix (ABM / synthetic cell-binding peptide P-15 in the formation of bone in human alveoli. Materials and methods: Eighteen patients in need of extraction of maxillary anterior teeth were selected and randomly assigned to the test group (ADM plus ABM/P-15 or the control group (ADM only. Histomorphometric measurements and histological analysis were recorded about 6 months after ridge preservation procedures in ten patients. The amount of newly formed bone, the most recently formed bone, fibrous tissue plus marrow spaces and remaining graft particles were measured and analyzed. Results: At 6 months, the new bone area parameter and the percentage of fibrous tissue plus marrow space areas showed higher values to the control group, and statistically significant differences when compared with the test group (p=0.03. Conclusion: The ADM acted as a membrane. The association of ABM/P-15 with ADM resulted in new bone formation within the alveoli, but the results were not considered relevant when used in this indication.

  8. The Use of Platelet Rich Plasma, Bone Morphogenetic Protein-2 and Different Scaffolds in Oral and Maxillofacial Surgery - Literature Review in Comparison with Own Clinical Experience

    OpenAIRE

    Karl-Heinz Schuckert; Stefan Jopp; Magdalena Osadnik

    2011-01-01

    ABSTRACT Objectives The purpose of this article was to review and critically assess the use of platelet rich plasma, recombinant human bone morphogenetic protein-2 and different scaffolds (i.e. tricalciumphosphate, polycaprolactone, demineralized bone matrix and anorganic bovine bone mineral) in oral and maxillofacial surgery comparing the relevant literature and own clinical experience. Material and Methods A literature review was conducted using MEDLINE, MEDPILOT and COCHRANE DATABASE OF SY...

  9. Pharmacological Inhibition of Protein Kinase G1 Enhances Bone Formation by Human Skeletal Stem Cells Through Activation of RhoA-Akt Signaling

    DEFF Research Database (Denmark)

    Kermani, Abbas Jafari; Siersbaek, Majken S; Chen, Li;

    2015-01-01

    for several malignant and nonmalignant conditions. We screened a library of kinase inhibitors to identify small molecules that enhance bone formation by human skeletal (stromal or mesenchymal) stem cells (hMSC). We identified H-8 (known to inhibit protein kinases A, C, and G) as a potent enhancer of...... differentiation and suggest that pharmacological inhibition of PRKG1 in hMSC implanted at the site of bone defect can enhance bone regeneration. Stem Cells 2015....

  10. Systematic identification of yeast proteins extracted into model wine during aging on the yeast lees.

    Science.gov (United States)

    Rowe, Jeffrey D; Harbertson, James F; Osborne, James P; Freitag, Michael; Lim, Juyun; Bakalinsky, Alan T

    2010-02-24

    Total protein and protein-associated mannan concentrations were measured, and individual proteins were identified during extraction into model wines over 9 months of aging on the yeast lees following completion of fermentations by seven wine strains of Saccharomyces cerevisiae. In aged wines, protein-associated mannan increased about 6-fold (+/-66%), while total protein only increased 2-fold (+/-20%), which resulted in a significantly greater protein-associated mannan/total protein ratio for three strains. A total of 219 proteins were identified among all wine samples taken over the entire time course. Of the 17 "long-lived" proteins detected in all 9 month samples, 13 were cell wall mannoproteins, and four were glycolytic enzymes. Most cytosolic proteins were not detected after 6 months. Native mannosylated yeast invertase was assayed for binding to wine tannin and was found to have a 10-fold lower affinity than nonglycosylated bovine serum albumin. Enrichment of mannoproteins in the aged model wines implies greater solution stability than other yeast proteins and the possibility that their contributions to wine quality may persist long after bottling. PMID:20108898

  11. Icariine stimulates proliferation and differentiation of human osteoblasts by increasing production of bone morphogenetic protein 2

    Institute of Scientific and Technical Information of China (English)

    YIN Xiao-xue; CHEN Zhong-qiang; LIU Zhong-jun; MA Qing-jun; DANG Geng-ting

    2007-01-01

    Background lcariine is a flavonoid isolated from a traditional Chinese medicine Epimedium pubescens and is the main active compound of it. Recently, Epimedium pubescens was found to have a therapeutic effect on osteoporosis. But the mechanism is unclear. The aim of the study was to research the effect of lcariine on the proliferation and differentiation of human osteoblasts.Methods Human osteoblasts were obtained byinducing human marrow mesenchymal stem cells (hMSCs) directionally and were cultured in the presence of various concentrations of lcariine. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was used to observe the effect of lcariine on cell proliferation. The activity of alkaline phosphatase (ALP) and the amount of calcified nodules were assayed to observe the effect on cell differentiation.The expression of bone morphogenetic protein 2 (BMP-2) mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR).Results Icariine (20 μg/ml) increased significantly the proliferation of human osteoblasts. And, lcariine (10 μg/ml and 20μg/ml) increased the activity of ALP and the amount of calcified nodules of human osteoblasts significantly (P<0.05).BMP-2 mRNA synthesis was elevated significantly in response to lcariine (20 μg/ml).Conclusions lcariine has a direct stimulatory effect on the proliferation and differentiation of cultured human osteoblastcells in vitro, which may be mediated by increasing production of BMP-2 in osteoblasts.

  12. Bone Morphogenetic Proteins stimulate mammary fibroblasts to promote mammary carcinoma cell invasion.

    Directory of Open Access Journals (Sweden)

    Philip Owens

    Full Text Available Bone Morphogenetic Proteins (BMPs are secreted cytokines that are part of the Transforming Growth Factor β (TGFβ superfamily. BMPs have been shown to be highly expressed in human breast cancers, and loss of BMP signaling in mammary carcinomas has been shown to accelerate metastases. Interestingly, other work has indicated that stimulation of dermal fibroblasts with BMP can enhance secretion of pro-tumorigenic factors. Furthermore, treatment of carcinoma-associated fibroblasts (CAFs derived from a mouse prostate carcinoma with BMP4 was shown to stimulate angiogenesis. We sought to determine the effect of BMP treatment on mammary fibroblasts. A large number of secreted pro-inflammatory cytokines and matrix-metallo proteases (MMPs were found to be upregulated in response to BMP4 treatment. Fibroblasts that were stimulated with BMP4 were found to enhance mammary carcinoma cell invasion, and these effects were inhibited by a BMP receptor kinase antagonist. Treatment with BMP in turn elevated pro-tumorigenic secreted factors such as IL-6 and MMP-3. These experiments demonstrate that BMP may stimulate tumor progression within the tumor microenvironment.

  13. Bone morphogenetic protein-2 is a negative regulator of hepatocyte proliferation downregulated in the regenerating liver

    Institute of Scientific and Technical Information of China (English)

    Cui-Ping Xu; Wen-Min Ji; Gijs R van den Brink; Maikel P Peppelenbosch

    2006-01-01

    AIM: To characterize the expression and dynamic changes of bone morphogenetic protein (BMP)-2 in hepatocytes in the regenerating liver in rats after partial hepatectomy (PH), and examine the effects of BMP-2 on proliferation of human Huh7 hepatoma cells.METHODS: Fifty-four adult male Wistar rats were randomly divided into three groups: A normal control (NC) group, a partial hepatectomized (PH) group and a sham operated (SO) group. To study the effect of liver regeneration on BMP-2 expression, rats were sacrificed before and at different time points after PH or the sham intervention (6, 12, 24 and 48 h). For each time point, six rats were used in parallel. Expression and distribution of BMP-2 protein were determined in regenerating liver tissue by Western blot analysis and immunohistochemistry. Effects of BMP-2 on cell proliferation of human Huh7 hepatoma cell line were assessed using an MTT assay.RESULTS: In the normal liver strong BMP-2 expression was observed around the central and portal veins. The expression of BMP-2 decreased rapidly as measured by both immunohistochemistry and Western blot analysis.This decrease was at a maximum of 3.22 fold after 12 h and returned to normal levels at 48 h after PH. No significant changes in BMP-2 immunoreactivity were observed in the SO group. BMP-2 inhibited serum induced Huh7 cell proliferation.CONCLUSION: BMP-2 is expressed in normal adult rat liver and negatively regulates hepatocyte proliferation.The observed down regulation of BMP-2 following partial hepatectomy suggests that such down regulation may be necessary for hepatocyte proliferation.

  14. Hypermethylation leads to bone morphogenetic protein 6 downregulation in hepatocellular carcinoma.

    Directory of Open Access Journals (Sweden)

    Yinghua He

    Full Text Available BACKGROUND: In the liver, bone morphogenetic protein 6 (BMP-6 maintains balanced iron metabolism. However, the mechanism that underlies greater BMP-6 expression in hepatocellular carcinoma (HCC tissue than adjacent non-cancerous tissue is unclear. This study sought to investigate the epigenetic mechanisms of BMP-6 expression by analysing the relationship between the DNA methylation status of BMP-6 and the expression of BMP-6. METHODS: Methylation-specific polymerase chain reaction (PCR, bisulphite sequencing PCR, the MethyLight assay, and quantitative real-time PCR were performed to examine BMP-6 methylation and mRNA expression levels. Immunohistochemistry (IHC was performed on tissue arrays to evaluate the BMP-6 protein level. RESULTS: BMP-6 mRNA expression was approximately 84.09% lower in HCC tissues than in adjacent non-cancerous tissues, and this low level of expression was associated with a poor prognosis. Moreover, the hypermethylation observed in HCC cell lines and HCC tissues was correlated with the BMP-6 mRNA expression level, and this correlation was validated following treatment with 5-aza-CdR, a demethylation agent. In addition, BMP-6 DNA methylation was upregulated by 68.42% in 114 clinical HCC tissue samples compared to adjacent normal tissues, whereas the BMP-6 staining intensity was downregulated by 77.03% in 75 clinical HCC tissue samples in comparison to adjacent normal tissues. Furthermore, elevated expression of BMP-6 in HCC cell lines inhibited cell colony formation. CONCLUSIONS: Our results suggest that BMP-6 CpG island hypermethylation leads to decreased BMP-6 expression in HCC tissues.

  15. Bone morphogenetic protein 15 may promote follicle selection in the hen.

    Science.gov (United States)

    Stephens, C S; Johnson, P A

    2016-09-01

    In the hen, optimal ovulation rate depends on selection of a single follicle into the pre-ovulatory hierarchy. Follicle selection is associated with increased oocyte growth and changes in gene expression in granulosa cells surrounding the oocyte, in preparation for ovulation. This study investigated the expression, function and regulation of bone morphogenetic protein-15 (BMP15) during follicle development in the hen. BMP15 mRNA expression was analyzed in the ooplasm and granulosa cells of 3mm follicles and was confirmed to be primarily in the ooplasm. BMP15 was detected by immunoblotting in 6 and 8mm follicles near the time of follicle selection. Expression of mRNA for BMP15 receptors (BMPR1B and BMPR2) in granulosa cells increased with follicle size, indicating that BMP15 may play an important role around follicle selection. The function of BMP15 was examined by culturing granulosa cells from 3-5mm and 6-8mm follicles with recombinant human BMP15 (rhBMP15). BMP15 increased expression of follicle stimulating hormone receptor (FSHR) mRNA and decreased anti-Müllerian hormone (AMH) mRNA and occludin (OCLN), factors associated with follicle maturation and growth in the hen. Hormonal regulation of BMP15 was assessed by whole follicle culture with estradiol (E2) which increased BMP15 mRNA expression. The distinct expression pattern of BMP15 and its receptors, coupled with the effects of BMP15 to increase FSHR mRNA and decrease AMH mRNA and OCLN mRNA and protein expression suggest that the oocyte may have a role in follicle selection in the chicken. PMID:27340039

  16. Rapid, room-temperature synthesis of amorphous selenium/protein composites using Capsicum annuum L extract

    Energy Technology Data Exchange (ETDEWEB)

    Li Shikuo; Shen Yuhua; Xie Anjian; Yu Xuerong; Zhang Xiuzhen; Yang Liangbao; Li Chuanhao [School of Chemistry and Chemical Engineering, Anhui University, Hefei 230039 (China)

    2007-10-10

    We describe the formation of amorphous selenium ({alpha}-Se)/protein composites using Capsicum annuum L extract to reduce selenium ions (SeO{sub 3}{sup 2-}) at room temperature. The reaction occurs rapidly and the process is simple and easy to handle. A protein with a molecular weight of 30 kDa extracted from Capsicum annuum L not only reduces the SeO{sub 3}{sup 2-} ions to Se{sup 0}, but also controls the nucleation and growth of Se{sup 0}, and even participates in the formation of {alpha}-Se/protein composites. The size and shell thickness of the {alpha}-Se/protein composites increases with high Capsicum annuum L extract concentration, and decreases with low reaction solution pH. The results suggest that this eco-friendly, biogenic synthesis strategy could be widely used for preparing inorganic/organic biocomposites. In addition, we also discuss the possible mechanism of the reduction of SeO{sub 3}{sup 2-} ions by Capsicum annuum L extract.

  17. Improvement of protein extraction from sunflower meal by hydrolysis with alcalase

    Directory of Open Access Journals (Sweden)

    Vioque, J.

    2003-12-01

    Full Text Available Extraction of proteins from defatted sunflower meal has been improved by addition of the protease alcalase during alkaline extraction. This method offers several additional advantages as compared to the traditional alkaline extraction without alcalase, which is usually carried out after a sedimentation/flotation step to remove the lignocellulosic fraction. As compared to extraction without alcalase, addition of 0.1% (v/v alcalase improved the yield of protein extraction from 57.5% to 87.4%, providing an extract that is 22% hydrolyzed. In addition, an increment of up to 4.5 times in protein solubility at low pH values is achieved, which correlates with the degree of hydrolysis. The extracts that were obtained in the presence of alcalase had a higher proline and glycine content, suggesting that the protease improves extraction of proline-rich and glycine-rich cell wall proteins that are part of the lignocellulosic fraction. These protein extracts can be directly dried without generation of wastewater, and the resulting fiber-rich material could be used for animal feeding.Se ha mejorado la extracción proteica de la harina desengrasa de girasol mediante la adición de la proteasa alcalasa durante la extracción alcalina. Este método ofrece varias ventajas adicionales en comparación con la extracción alcalina tradicional sin alcalasa, que se desarrolla normalmente mediante un proceso de flotación/sedimentación para retirar la fracción lignocelulósica. En comparación a la extracción sin alcalasa, la adicción de 0.1% (v/v de alcalasa mejora los rendimientos de extracción proteica desde un 57.5% a un 87.4%, dando un extracto con un 22% de grado de hidrólisis. Además se obtiene un incremento de hasta 4.5 veces de la solubilidad proteica a bajos pHs, que se correlaciona con el grado de hidrólisis. Los extractos obtenidos con alcalasa tenían un mayor contenido de prolina y glicina, sugiriendo que la proteasa mejora la extracción de las

  18. Effects of supercritical fluid extraction conditions on yield of protein from defatted rice bran

    International Nuclear Information System (INIS)

    Summary: In the present study, effect of different supercritical fluid extractor (SFE) operational conditions i.e. pressure (300, 450, 600 bars), flow rate (10.0, 17.5, 25.0 g/min), and intervals of extraction (30, 90, 150 min) at 60 degree C on protein content of defatted rice bran (DRB) samples, has been evaluated. The DRB samples were collected from SFE vessel and analyzed for their moisture, residual fat and protein contents. The protein content was determined by Kjeldahl method and ranged over 15.59 to 17.44%, while moisture content was determined by drying method and found to be ranging from 7.84 to 9.88%, and residual fat content was determined by Soxhlet method and varied over 4.67 to 18.94%. Differences were selectively significant (p > 0.05) among the protein, residual fat, and moisture contents, as function of extraction conditions. Highest yield of protein was obtained at 450 bars, 90 min extraction time, and 17.5 g/min flow rate, while the lowest at 300 bars, 30 min of extraction time, and 10.0 g/min flow rate. Negative correlation with p-value less than or equal to 0.05 was found between moisture content and residual fat content of DRB. However, no correlation could be observed among protein, moisture and residual fat contents of DRB, respectively. The study demonstrates that SFE conditions have non-significant impact on the yield of protein in defatted materials. (author)

  19. Screening for carbohydrate-binding proteins in extracts of Uruguayan plants

    Directory of Open Access Journals (Sweden)

    Plá A.

    2003-01-01

    Full Text Available The presence of carbohydrate-binding proteins, namely lectins, ß-galactosidases and amylases, was determined in aqueous extracts of plants collected in Uruguay. Twenty-six extracts were prepared from 15 Uruguayan plants belonging to 12 Phanerogam families. Among them, 18 extracts caused hemagglutination (HAG that was inhibited by mono- and disaccharides in 13 cases, indicating the presence of lectins. The other 8 extracts did not cause any HAG with the four systems used to detect HAG activity (rabbit and mouse red cells, trypsin-treated rabbit and mouse red cells. For the extracts prepared from Solanum commersonii, HAG activity and HAG inhibition were similar for those prepared from tubers, leaves and fruits, with the chitocompounds being responsible for all the inhibitions. Purification of the S. commersonii tuber lectin was carried out by affinity chromatography on asialofetuin-Sepharose, and SDS-PAGE under reducing conditions gave a single band of Mr of approximately 80 kDa. The monomer N-acetylglucosamine did not inhibit HAG induced by the purified lectin, but chitobiose inhibited HAG at 24 mM and chitotriose inhibited it at 1 mM. ß-Galactosidase activity was detected in leaves and stems of Cayaponia martiana, and in seeds from Datura ferox. Only traces of amylase activity were detected in some of the extracts analyzed. The present screening increases knowledge about the occurrence of carbohydrate-binding proteins present in regional plants.

  20. The Use of Platelet Rich Plasma, Bone Morphogenetic Protein-2 and Different Scaffolds in Oral and Maxillofacial Surgery - Literature Review in Comparison with Own Clinical Experience

    Directory of Open Access Journals (Sweden)

    Karl-Heinz Schuckert

    2011-01-01

    Full Text Available Objectives: The purpose of this article was to review and critically assess the use of platelet rich plasma, recombinant human bone morphogenetic protein-2 and different scaffolds (i.e. tricalciumphosphate, polycaprolactone, demineralized bone matrix and anorganic bovine bone mineral in oral and maxillofacial surgery comparing the relevant literature and own clinical experience.Material and Methods: A literature review was conducted using MEDLINE, MEDPILOT and COCHRANE DATABASE OF SYSTEMATIC REVIEWS. It concentrated on manuscripts and overviews published in the last five years (2006-2010. The key terms employed were platelet rich plasma, bone morphogenetic proteins and their combinations with the above mentioned scaffolds. The results of clinical studies and animal trials were especially emphasized. The statements from the literature were compared with authors’ own clinical data.Results: New publications and overviews demonstrate the advantages of platelet rich plasma in bone regeneration. The results from the literature review were discussed and compared with the publications detailing authors’ own experiences.Conclusions: A favourable outcome concerning newly grown bone was achieved combining platelet rich plasma in addition to optimal matrices with or without recombinant human bone morphogenetic protein-2, depending on the clinical case. As a consequence, the paradigm shift from transplantation of autogenous bone to bone tissue engineering appears promising.

  1. Injectable calcium phosphate cement and fibrin sealant recombined human bone morphogenetic protein-2 composite in vertebroplasty: an animal study

    OpenAIRE

    Qian, Guang; Dong, Youhai; Yang, Wencheng; Wang, Minghai

    2012-01-01

    Polymethylmethacrylate (PMMA) is currently the most commonly-used material, but it may induce adjacent vertebral fracture due to low degradation and high strength. Our study evaluated the feasibility of injectable calcium phosphate cement (ICPC) and fibrin sealant (FS) as an injectable compound carrier of human bone morphogenetic protein-2 (rhBMP-2) in New Zealand rabbits for vertebroplasty. Results showed ICPC/FS/rhBMP-2 composites induced alkaline phosphatase most effectively at 2 and 4 wee...

  2. Bone Morphogenetic Protein-2-Induced Signaling and Osteogenesis Is Regulated by Cell Shape, RhoA/ROCK, and Cytoskeletal Tension

    OpenAIRE

    Wang, Yang-Kao; Yu, Xiang; Cohen, Daniel M.; Wozniak, Michele A.; Yang, Michael T.; Gao, Lin; Eyckmans, Jeroen; Chen, Christopher S.

    2011-01-01

    Osteogenic differentiation of human mesenchymal stem cells (hMSCs) is classically thought to be mediated by different cytokines such as the bone morphogenetic proteins (BMPs). Here, we report that cell adhesion to extracellular matrix (ECM), and its effects on cell shape and cytoskeletal mechanics, regulates BMP-induced signaling and osteogenic differentiation of hMSCs. Using micropatterned substrates to progressively restrict cell spreading and flattening against ECM, we demonstrated that BM...

  3. Metformin revisited: Does this regulator of AMP-activated protein kinase secondarily affect bone metabolism and prevent diabetic osteopathy

    OpenAIRE

    McCarthy, Antonio Desmond; Cortizo, Ana María; Sedlinsky, Claudia

    2016-01-01

    Patients with long-term type 1 and type 2 diabetes mellitus (DM) can develop skeletal complications or “diabetic osteopathy”. These include osteopenia, osteoporosis and an increased incidence of low-stress fractures. In this context, it is important to evaluate whether current anti-diabetic treatments can secondarily affect bone metabolism. Adenosine monophosphate-activated protein kinase (AMPK) modulates multiple metabolic pathways and acts as a sensor of the cellular energy status; recent e...

  4. The bone morphogenetic protein receptor-1A pathway is required for lactogenic differentiation of mammary epithelial cells in vitro

    OpenAIRE

    Perotti, C.; Karayazi, Ö.; Moffat, S.; Shemanko, C. S.

    2012-01-01

    Bone morphogenetic proteins (BMPs) have been implicated in the control of proliferation, tissue formation, and differentiation. BMPs regulate the biology of stem and progenitor cells and can promote cellular differentiation, depending on the cell type and context. Although the BMP pathway is known to be involved in early embryonic development of the mammary gland via mesenchymal cells, its role in later epithelial cellular differentiation has not been examined. The majority of the mammary gla...

  5. Extracting protein dynamics information from overlapped NMR signals using relaxation dispersion difference NMR spectroscopy

    International Nuclear Information System (INIS)

    Protein dynamics plays important roles in many biological events, such as ligand binding and enzyme reactions. NMR is mostly used for investigating such protein dynamics in a site-specific manner. Recently, NMR has been actively applied to large proteins and intrinsically disordered proteins, which are attractive research targets. However, signal overlap, which is often observed for such proteins, hampers accurate analysis of NMR data. In this study, we have developed a new methodology called relaxation dispersion difference that can extract conformational exchange parameters from overlapped NMR signals measured using relaxation dispersion spectroscopy. In relaxation dispersion measurements, the signal intensities of fluctuating residues vary according to the Carr-Purcell-Meiboon-Gill pulsing interval, whereas those of non-fluctuating residues are constant. Therefore, subtraction of each relaxation dispersion spectrum from that with the highest signal intensities, measured at the shortest pulsing interval, leaves only the signals of the fluctuating residues. This is the principle of the relaxation dispersion difference method. This new method enabled us to extract exchange parameters from overlapped signals of heme oxygenase-1, which is a relatively large protein. The results indicate that the structural flexibility of a kink in the heme-binding site is important for efficient heme binding. Relaxation dispersion difference requires neither selectively labeled samples nor modification of pulse programs; thus it will have wide applications in protein dynamics analysis

  6. Extracting protein dynamics information from overlapped NMR signals using relaxation dispersion difference NMR spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Konuma, Tsuyoshi [Icahn School of Medicine at Mount Sinai, Department of Structural and Chemical Biology (United States); Harada, Erisa [Suntory Foundation for Life Sciences, Bioorganic Research Institute (Japan); Sugase, Kenji, E-mail: sugase@sunbor.or.jp, E-mail: sugase@moleng.kyoto-u.ac.jp [Kyoto University, Department of Molecular Engineering, Graduate School of Engineering (Japan)

    2015-12-15

    Protein dynamics plays important roles in many biological events, such as ligand binding and enzyme reactions. NMR is mostly used for investigating such protein dynamics in a site-specific manner. Recently, NMR has been actively applied to large proteins and intrinsically disordered proteins, which are attractive research targets. However, signal overlap, which is often observed for such proteins, hampers accurate analysis of NMR data. In this study, we have developed a new methodology called relaxation dispersion difference that can extract conformational exchange parameters from overlapped NMR signals measured using relaxation dispersion spectroscopy. In relaxation dispersion measurements, the signal intensities of fluctuating residues vary according to the Carr-Purcell-Meiboon-Gill pulsing interval, whereas those of non-fluctuating residues are constant. Therefore, subtraction of each relaxation dispersion spectrum from that with the highest signal intensities, measured at the shortest pulsing interval, leaves only the signals of the fluctuating residues. This is the principle of the relaxation dispersion difference method. This new method enabled us to extract exchange parameters from overlapped signals of heme oxygenase-1, which is a relatively large protein. The results indicate that the structural flexibility of a kink in the heme-binding site is important for efficient heme binding. Relaxation dispersion difference requires neither selectively labeled samples nor modification of pulse programs; thus it will have wide applications in protein dynamics analysis.

  7. PIPE: a protein–protein interaction passage extraction module for BioCreative challenge

    Science.gov (United States)

    Chu, Chun-Han; Su, Yu-Chen; Chen, Chien Chin; Hsu, Wen-Lian

    2016-01-01

    Identifying the interactions between proteins mentioned in biomedical literatures is one of the frequently discussed topics of text mining in the life science field. In this article, we propose PIPE, an interaction pattern generation module used in the Collaborative Biocurator Assistant Task at BioCreative V (http://www.biocreative.org/) to capture frequent protein-protein interaction (PPI) patterns within text. We also present an interaction pattern tree (IPT) kernel method that integrates the PPI patterns with convolution tree kernel (CTK) to extract PPIs. Methods were evaluated on LLL, IEPA, HPRD50, AIMed and BioInfer corpora using cross-validation, cross-learning and cross-corpus evaluation. Empirical evaluations demonstrate that our method is effective and outperforms several well-known PPI extraction methods. Database URL: PMID:27524807

  8. Cell-free protein expression based on extracts from CHO cells.

    Science.gov (United States)

    Brödel, Andreas K; Sonnabend, Andrei; Kubick, Stefan

    2014-01-01

    Protein expression systems are widely used in biotechnology and medicine for the efficient and economic production of therapeutic proteins. Today, cultivated Chinese hamster ovary (CHO) cells are the market dominating mammalian cell-line for the production of complex therapeutic proteins. Despite this outstanding potential of CHO cells, no high-yield cell-free system based on translationally active lysates from these cells has been reported so far. To date, CHO cell extracts have only been used as a foundational research tool for understanding mRNA translation (Lodish et al., 1974; McDowell et al., 1972). In the present study, we address this fact by establishing a novel cell-free protein expression system based on extracts from cultured CHO cells. Lysate preparation, adaptation of in vitro reaction conditions and the construction of particular expression vectors are considered for high-yield protein production. A specific in vitro expression vector, which includes an internal ribosome entry site (IRES) from the intergenic region (IGR) of the Cricket paralysis virus (CrPV), has been constructed in order to obtain optimal performance. The IGR IRES is supposed to bind directly to the eukaryotic 40S ribosomal subunit thereby bypassing the process of translation initiation, which is often a major bottleneck in cell-free systems. The combination of expression vector and optimized CHO cell extracts enables the production of approximately 50 µg/mL active firefly luciferase within 4 h. The batch-type cell-free coupled transcription-translation system has the potential to perform post-translational modifications, as shown by the glycosylation of erythropoietin. Accordingly, the system contains translocationally active endogenous microsomes, enabling the co-translational incorporation of membrane proteins into biological membranes. Hence, the presented in vitro translation system is a powerful tool for the fast and convenient optimization of expression constructs, the

  9. A continuum of research projects to improve extraction of oil and proteins in oilseed plants

    Directory of Open Access Journals (Sweden)

    Miquel Martine

    2011-05-01

    Full Text Available A key challenge in the actual context of fossil sources rarefaction, global warming, and of increase of the world global population, is to promote the use of molecules derived from renewable sources such as plants. Among these molecules, lipids and proteins are targets of interest. Plant lipids from oilseeds are attractive substitutes to the use of fossil oil. Till the beginning of the 20th century, numerous products used in the daily life were derived from natural renewable products. For instance, plant oil was commonly used as fuel for vehicles and was entering in the composition of paintings, lubricants etc. Unfortunately, natural oils have been progressively replaced by cheaper fossil oil in the fabrication of these products. Nowadays, fossil oils are becoming increasingly expensive being a finite comodity. It is thus important to reduce our dependence from fossil oil and develop substitution industries. Oilseeds contain important amounts of proteins which are mainly used in feed. As several kilograms of plant protein are needed to obtain one kilogram of animal protein, the interest toward using plant protein in food is reinforced. The developments of the use of plant lipids, as well as proteins are a major stakes for the competitiveness of European agriculture and industry, as well as for sustainable development. Extraction of oil and proteins from rapeseed has a significant cost, in term of energy and solvent uses, and finally affects the ultimate quality of the products (protein digestibility. In order to quantitatively extract seed reserves under mild conditions, it will be necessary to limit the amount of energy needed, and avoid any use of solvents. Ideally, seeds should be processed in a bio refinery. In this paper, we will describe how oilseeds store their reserves, and roadblocks for improving actual oilseed extraction processes. A continuum of research projects aimed at answering targeted questions will be presented, with selected

  10. Facilitated receptor-recognition and enhanced bioactivity of bone morphogenetic protein-2 on magnesium-substituted hydroxyapatite surface

    Science.gov (United States)

    Huang, Baolin; Yuan, Yuan; Li, Tong; Ding, Sai; Zhang, Wenjing; Gu, Yuantong; Liu, Changsheng

    2016-04-01

    Biomaterial surface functionalized with bone morphogenetic protein-2 (BMP-2) is a promising approach to fabricating successful orthopedic implants/scaffolds. However, the bioactivity of BMP-2 on material surfaces is still far from satisfactory and the mechanism of related protein-surface interaction remains elusive. Based on the most widely used bone-implants/scaffolds material, hydroxyapatite (HAP), we developed a matrix of magnesium-substituted HAP (Mg-HAP, 2.2 at% substitution) to address these issues. Further, we investigated the adsorption dynamics, BMPRs-recruitment, and bioactivity of recombinant human BMP-2 (rhBMP-2) on the HAP and Mg-HAP surfaces. To elucidate the mechanism, molecular dynamic simulations were performed to calculate the preferred orientations, conformation changes, and cysteine-knot stabilities of adsorbed BMP-2 molecules. The results showed that rhBMP-2 on the Mg-HAP surface exhibited greater bioactivity, evidenced by more facilitated BMPRs-recognition and higher ALP activity than on the HAP surface. Moreover, molecular simulations indicated that BMP-2 favoured distinct side-on orientations on the HAP and Mg-HAP surfaces. Intriguingly, BMP-2 on the Mg-HAP surface largely preserved the active protein structure evidenced by more stable cysteine-knots than on the HAP surface. These findings explicitly clarify the mechanism of BMP-2-HAP/Mg-HAP interactions and highlight the promising application of Mg-HAP/BMP-2 matrixes in bone regeneration implants/scaffolds.

  11. The synergistic induction of bone formation by the osteogenic proteins of the TGF-β supergene family.

    Science.gov (United States)

    Ripamonti, Ugo; Parak, Ruqayya; Klar, Roland M; Dickens, Caroline; Dix-Peek, Thérèse; Duarte, Raquel

    2016-10-01

    The momentum to compose this Leading Opinion on the synergistic induction of bone formation suddenly arose when a simple question was formulated during a discussion session on how to boost the often limited induction of bone formation seen in clinical contexts. Re-examination of morphological and molecular data available on the rapid induction of bone formation by the recombinant human transforming growth factor-β3 (hTGF-β3) shows that hTGF-β3 replicates the synergistic induction of bone formation as invocated by binary applications of hOP-1:hTGF-β1 at 20:1 by weight when implanted in heterotopic sites of the rectus abdominis muscle of the Chacma baboon, Papio ursinus. The rapid induction of bone formation in primates by hTGF-β3 may stem from bursts of cladistic evolution, now redundant in lower animal species but still activated in primates by relatively high doses of hTGF-β3. Contrary to rodents, lagomorphs and canines, the three mammalian TGF-β isoforms induce rapid and substantial bone formation when implanted in heterotopic rectus abdominis muscle sites of P. ursinus, with unprecedented regeneration of full thickness mandibular defects with rapid mineralization and corticalization. Provocatively, thus providing potential molecular and biological rationales for the apparent redundancy of osteogenic molecular signals in primates, binary applications of recombinant human osteogenic protein-1 (hOP-1) with low doses of hTGF-β1 and -β3, synergize to induce massive ossicles in heterotopic rectus abdominis, orthotopic calvarial and mandibular sites of P. ursinus. The synergistic binary application of homologous but molecularly different soluble molecular signals has indicated that per force several secreted molecular signals are required singly, synchronously and synergistically to induce optimal osteogenesis. The morphological hallmark of the synergistic induction of bone formation is the rapid differentiation of large osteoid seams enveloping

  12. Electrophoretic Extraction and Proteomic Characterization of Proteins Buried in Marine Sediments

    Directory of Open Access Journals (Sweden)

    Eli K. Moore

    2014-10-01

    Full Text Available Proteins are the largest defined molecular component of marine organic nitrogen, and hydrolysable amino acids, the building blocks of proteins, are important components of particulate nitrogen in marine sediments. In oceanic systems, the largest contributors are phytoplankton proteins, which have been tracked from newly produced bloom material through the water column to surface sediments in the Bering Sea, but it is not known if proteins buried deeper in sediment systems can be identified with confidence. Electrophoretic gel protein extraction methods followed by proteomic mass spectrometry and database searching were used as the methodology to identify buried phytoplankton proteins in sediments from the 8–10 cm section of a Bering Sea sediment core. More peptides and proteins were identified using an SDS-PAGE tube gel than a standard 1D flat gel or digesting the sediment directly with trypsin. The majority of proteins identified correlated to the marine diatom, Thalassiosira pseudonana, rather than bacterial protein sequences, indicating an algal source not only dominates the input, but also the preserved protein fraction. Abundant RuBisCO and fucoxanthin chlorophyll a/c binding proteins were identified, supporting algal sources of these proteins and reinforcing the proposed mechanisms that might protect proteins for long time periods. Some preserved peptides were identified in unexpected gel molecular weight ranges, indicating that some structural changes or charge alteration influenced the mobility of these products during electrophoresis isolation. Identifying buried photosystem proteins suggests that algal particulate matter is a significant fraction of the preserved organic carbon and nitrogen pools in marine sediments.

  13. Acetate-dependent methylation of two corrinoid proteins in extracts of Methanosarcina barkeri

    International Nuclear Information System (INIS)

    Corrinoid proteins have been implicated as methyl carriers in methane formation from acetate, yet specific corrinoid proteins methylated by acetate-derived intermediates have not been identified. In the presence of ATP, H2, and bromoethanesulfonic acid, label from 3H- or 2-14C-labeled acetate was incorporated into the protein fraction of cell extracts of Methanosarcina barkeri. Incorporated label was susceptible to photolysis, yielding labeled methane as the anaerobic photolysis product. size exclusion high-pressure liquid chromatography (HPLC) demonstrated the presence of at least three labeled proteins with native molecular sizes of 480, 200, and 29 kDa, while electrophoresis indicated that four major labeled proteins were present. Dual-label experiments demonstrated that these four proteins were methylated rather than acetylated. Two of the proteins (480 and 29 kDa) contained the majority of radiolabel and were stably methylated. After labeling with [2-14C]acetate, the stable 14CH3-proteins were partially purified, and 14CH3-cofactors were isolated from each protein. UV-visible spectroscopy and HPLC demonstrated these to be methylated corrinoids. When the protein. UV-visible spectroscopy and HPLC demonstrated these to be methylated corrinoids. When the 480-kDa corrinoid protein was purified to 70% homogeneity, the preparation was found to have subunits of 40 and 30 kDa. The 480-kDa protein but not the 29-kDa protein was methylated during in vitro methanogenesis from acetate and demethylated as methanogenesis ceased, consistent with the involvement of this protein in methane formation

  14. Osteoblast Differentiation and Bone: Relevant proteins, regulatory processes and the vascular connection

    NARCIS (Netherlands)

    R.D.A.M. Alves (Rodrigo)

    2012-01-01

    textabstractBone is a highly specialized form of connective tissue present in most vertebrate animals as part of the endoskeleton. Structurally speaking, bone is mainly constituted by an organic extracellular matrix (ECM) hardened by deposited mineral. The blending between the organic and inorganic

  15. In vivo screening of five phytochemicals/extracts and a fungal immunomodulatory protein against colibacillosis in broilers

    NARCIS (Netherlands)

    Peek, H.W.; Halkes, S.B.A.; Tomassen, M.M.M.; Mes, J.J.; Landman, W.J.M.

    2013-01-01

    Five phytochemicals/extracts (an extract from Echinacea purpurea, a ß-glucan-rich extract from Shiitake, betaine [Betain™], curcumin from Curcuma longa [turmeric] powder, carvacrol and also a recombinant fungal immunomodulatory protein [FIP] from Ganoderma lucidum) cloned and expressed in Escherichi

  16. Effect of Bovine Plasma Protein on Autolysis and Gelation of Protein Extracted from Giant Squid (Dosidicus gigas Mantle

    Directory of Open Access Journals (Sweden)

    Laura Raquel Marquez-Alvarez

    2015-01-01

    Full Text Available The effect of bovine plasma protein (BPP on the inhibition of autolytic activity and its effect on the gelling properties of a protein concentrate (PC obtained from jumbo squid (Dosidicus gigas mantle were investigated. Sols and gels were prepared from the PC by adding different amounts of BPP (0, 1, and 2%. Dynamic oscillatory measurements indicated that systems with 1% BPP had a higher elastic modulus (G′, in which hydrophobic interactions were favored. Concerning the technological and textural quality of the gels, BPP caused a greater water holding capacity (WHC, force, cohesiveness, and elasticity, probably due to improvement of the electrostatic and hydrophobic interactions during gel formation. Scanning electron microscopy (SEM allowed visualization of the formation of more rigid and ordered gels with less porosity when BPP was added. Therefore, the addition of BPP improved the gelling capacity of proteins extracted from giant squid.

  17. Determination of protein carbonyls in plasma, cell extracts, tissue homogenates, isolated proteins

    DEFF Research Database (Denmark)

    Weber, Daniela; Davies, Michael J.; Grune, Tilman

    2015-01-01

    the most relevant methods to detect protein carbonyls after derivatization with 2,4-dinitrophenylhydrazine with an emphasis on measurement in plasma, cells, organ homogenates, isolated proteins and organelles. Sample preparation, derivatization conditions and protein handling are presented for the...... different reactive oxygen species in blood, tissues and cells. Sample preparation and stabilization are key steps in the accurate quantification of oxidation-related products and examination of physiological/pathological processes. This review therefore focuses on the sample preparation processes used in...... spectrophotometric and HPLC method as well as for immunoblotting and ELISA. An extensive overview covering these methods in previously published articles is given for researchers who plan to measure protein carbonyls in different samples....

  18. Effect of xenon arc irradiation on mink hair protein extracted with thioglycolate

    International Nuclear Information System (INIS)

    The effect of xenon arc irradiation on the extractability and properties of mink hair protein treated with sodium thioglycolate solution was investigated. Over 50% of the intact sapphire mink hair was dissolved when treated with TG 11 (0.1M sodium thioglycolate, pH 11) and TG/G-7 (0.1M sodium thioglycolate, 6M guanidine-HCI, pH 7), but hardly dissolved with TG/U-7 (0.1M sodium thioglycolate, 7M urea, pH 7). The extractability of underfur was higher than that of guard hair with TG 11 or TG/G-7. The extractability with TG 11 or TG/G-7 was remarkably decreased in intact hair than irradiated hair. This implies that the irradiation-induced deterioration of mink hair involves the formation of some additional intermolecular or interpeptide crosslinks other than the original disulfide bridges in native hair. Amino acid analysis, SDS-PAGE and size exclusion chromatography of TG/G-7 extract revealed that the irradiation of mink hair made preferentially microfibril protein insoluble, and leaves matrix protein especially high-glycine-tyrosine component soluble

  19. The Ginkgo biloba Extract EGb 761 Modulates Proteasome Activity and Polyglutamine Protein Aggregation

    Directory of Open Access Journals (Sweden)

    Marcel Stark

    2014-01-01

    Full Text Available The standardized Ginkgo biloba extract EGb 761 has well-described antioxidative activities and effects on different cytoprotective signaling pathways. Consequently, a potential use of EGb 761 in neurodegenerative diseases has been proposed. A common characteristic feature of a variety of such disorders is the pathologic formation of protein aggregates, suggesting a crucial role for protein homeostasis. In this study, we show that EGb 761 increased the catalytic activity of the proteasome and enhanced protein degradation in cultured cells. We further investigated this effect in a cellular model of Huntington’s disease (HD by employing cells expressing pathologic variants of a polyglutamine protein (polyQ protein. We show that EGb 761 affected these cells by (i increasing proteasome activity and (ii inducing a more efficient degradation of aggregation-prone proteins. These results demonstrate a novel activity of EGb 761 on protein aggregates by enhancing proteasomal protein degradation, suggesting a therapeutic use in neurodegenerative disorders with a disturbed protein homeostasis.

  20. Automated data extraction from in situ protein stable isotope probing studies

    Energy Technology Data Exchange (ETDEWEB)

    Slysz, Gordon W.; Steinke, Laurey A.; Ward, David M.; Klatt, Christian G.; Clauss, Therese RW; Purvine, Samuel O.; Payne, Samuel H.; Anderson, Gordon A.; Smith, Richard D.; Lipton, Mary S.

    2014-01-27

    Protein stable isotope probing (protein-SIP) has strong potential for revealing key metabolizing taxa in complex microbial communities. While most protein-SIP work to date has been performed under controlled laboratory conditions to allow extensive isotope labeling of the target organism, a key application will be in situ studies of microbial communities under conditions that result in small degrees of partial labeling. One hurdle restricting large scale in situ protein-SIP studies is the lack of algorithms and software for automated data processing of the massive data sets resulting from such studies. In response, we developed Stable Isotope Probing Protein Extraction Resources software (SIPPER) and applied it for large scale extraction and visualization of data from short term (3 h) protein-SIP experiments performed in situ on Yellowstone phototrophic bacterial mats. Several metrics incorporated into the software allow it to support exhaustive analysis of the complex composite isotopic envelope observed as a result of low amounts of partial label incorporation. SIPPER also enables the detection of labeled molecular species without the need for any prior identification.

  1. Alkaline Extraction of Cobia (Rachycentroncanadum Proteins: Physicochemical characteristics, functional and thermal properties

    Directory of Open Access Journals (Sweden)

    Sabrina de La Rocha

    2015-04-01

    Full Text Available Cobia (Rachycentroncanadum is an important emerging species in Brazilian mariculture. The aim of the study was to obtain and characterize a protein isolate from cobia muscle using chemical extraction process by alkaline solubilization and isoelectric precipitation of proteins. The extraction yield was 98.17g/100g protein on a dry basis. The highest solubility and water holding capacity (WHC of cobiaprotein isolate (CPI was obtained at pH 11and 21.9mL of water per gram of protein. The electrophoretic profiles revealed masses characteristic of myofibrillar proteins (myosin and actin. The main peaks identified by qualitative spectroscopy analysis of the infrared spectrawere characteristic of peptide bonds such as amide I and amide II. The highest fusion and degradation points of CPI were 259.1°C and 378°C, respectively. The results showed that the CPI has great biotechnological value in various industrial areas that require a product of high protein value.

  2. A Mineral-Rich Extract from the Red Marine Algae Lithothamnion calcareum Preserves Bone Structure and Function in Female Mice on a Western-Style Diet

    OpenAIRE

    Aslam, Muhammad Nadeem; Kreider, Jaclynn M.; Paruchuri, Tejaswi; Bhagavathula, Narasimharao; DaSilva, Marissa; Zernicke, Ronald F.; Goldstein, Steven A.; Varani, James

    2010-01-01

    The purpose of this study was to determine whether a mineral-rich extract derived from the red marine algae Lithothamnion calcareum could be used as a dietary supplement for prevention of bone mineral loss. Sixty C57BL/6 mice were divided into three groups based on diet: the first group received a high-fat Western-style diet (HFWD), the second group was fed the same HFWD along with the mineral-rich extract included as a dietary supplement, and the third group was used as a control and was fed...

  3. Inhibitory Smads and bone morphogenetic protein (BMP) modulate anterior photoreceptor cell number during planarian eye regeneration.

    Science.gov (United States)

    González-Sastre, Alejandro; Molina, Ma Dolores; Saló, Emili

    2012-01-01

    Planarians represent an excellent model to study the processes of body axis and organ re-specification during regeneration. Previous studies have revealed a conserved role for the bone morphogenetic protein (BMP) pathway and its intracellular mediators Smad1/5/8 and Smad4 in planarian dorsoventral (DV) axis re-establishment. In an attempt to gain further insight into the role of this signalling pathway in planarians, we have isolated and functionally characte-rized the inhibitory Smads (I-Smads) in Schmidtea mediterranea. Two I-Smad homologues have been identified: Smed-smad6/7-1 and Smed-smad6/7-2. Expression of smad6/7-1 was detected in the parenchyma, while smad6/7-2 was found to be ex-pressed in the central nervous system and the eyes. Neither single smad6/7-1 and smad6/7-2 nor double smad6/7-1,-2 silencing gave rise to any apparent disruption of the DV axis. However, both regenerating and intact smad6/7-2 (RNAi) planarians showed defects in eye morphogenesis and displayed small, rounded eyes that lacked the anterior subpopulation of photoreceptor cells. The number of pigment cells was also reduced in these animals at later stages of regeneration. In contrast, after low doses of Smed-bmp(RNAi), planarians regenerated larger eyes in which the anterior subpopulation of photoreceptor cells was expanded. Our results suggest that Smed-smad6/7-2 and Smed-bmp control the re-specification and maintenance of anterior photoreceptor cell number in S. mediterranea. PMID:22451003

  4. Bone morphogenetic protein 15 in the pro-mature complex form enhances bovine oocyte developmental competence.

    Directory of Open Access Journals (Sweden)

    Jaqueline Sudiman

    Full Text Available Developmental competence of in vitro matured (IVM oocytes needs to be improved and this can potentially be achieved by adding recombinant bone morphogenetic protein 15 (BMP15 or growth differentiation factor (GDF9 to IVM. The aim of this study was to determine the effect of a purified pro-mature complex form of recombinant human BMP15 versus the commercially available bioactive forms of BMP15 and GDF9 (both isolated mature regions during IVM on bovine embryo development and metabolic activity. Bovine cumulus oocyte complexes (COCs were matured in vitro in control medium or treated with 100 ng/ml pro-mature BMP15, mature BMP15 or mature GDF9 +/- FSH. Metabolic measures of glucose uptake and lactate production from COCs and autofluorescence of NAD(PH, FAD and GSH were measured in oocytes after IVM. Following in vitro fertilisation and embryo culture, day 8 blastocysts were stained for cell numbers. COCs matured in medium +/- FSH containing pro-mature BMP15 displayed significantly improved blastocyst development (57.7±3.9%, 43.5±4.2% compared to controls (43.3±2.4%, 28.9±3.7% and to mature GDF9+FSH (36.1±3.0%. The mature form of BMP15 produced intermediate levels of blastocyst development; not significantly different to control or pro-mature BMP15 levels. Pro-mature BMP15 increased intra-oocyte NAD(PH, and reduced glutathione (GSH levels were increased by both forms of BMP15 in the absence of FSH. Exogenous BMP15 in its pro-mature form during IVM provides a functional source of oocyte-secreted factors to improve bovine blastocyst development. This form of BMP15 may prove useful for improving cattle and human artificial reproductive technologies.

  5. Failure of bone morphogenetic protein receptor trafficking in pulmonary arterial hypertension: potential for rescue.

    Science.gov (United States)

    Sobolewski, Anastasia; Rudarakanchana, Nung; Upton, Paul D; Yang, Jun; Crilley, Trina K; Trembath, Richard C; Morrell, Nicholas W

    2008-10-15

    Heterozygous germline mutations in the gene encoding the bone morphogenetic protein type II receptor cause familial pulmonary arterial hypertension (PAH). We previously demonstrated that the substitution of cysteine residues in the ligand-binding domain of this receptor prevents receptor trafficking to the cell membrane. Here we demonstrate the potential for chemical chaperones to rescue cell-surface expression of mutant BMPR-II and restore function. HeLa cells were transiently transfected with BMPR-II wild type or mutant (C118W) receptor constructs. Immunolocalization studies confirmed the retention of the cysteine mutant receptor mainly in the endoplasmic reticulum. Co-immunoprecipitation studies of Myc-tagged BMPR-II confirmed that the cysteine-substituted ligand-binding domain mutation, C118W, is able to associate with BMP type I receptors. Furthermore, following treatment with a panel of chemical chaperones (thapsigargin, glycerol or sodium 4-phenylbutyrate), we demonstrated a marked increase in cell-surface expression of mutant C118W BMPR-II by FACS analysis and confocal microscopy. These agents also enhanced the trafficking of wild-type BMPR-II, though to a lesser extent. Increased cell-surface expression of mutant C118W BMPR-II was associated with enhanced Smad1/5 phosphorylation in response to BMPs. These findings demonstrate the potential for rescue of mutant BMPR-II function from the endoplasmic reticulum. For the C118W mutation in the ligand-binding domain of BMPR-II, cell-surface rescue leads to at least partial restoration of BMP signalling. We conclude that enhancement of cell-surface trafficking of mutant and wild-type BMPR-II may have therapeutic potential in familial PAH. PMID:18647753

  6. Bone marrow mononuclear cells up-regulate toll-like receptor expression and produce inflammatory mediators in response to cigarette smoke extract.

    Directory of Open Access Journals (Sweden)

    Junmin Zhou

    Full Text Available Several reports link cigarette smoking with leukemia. However, the effects of cigarette smoke extract (CSE on bone marrow hematopoiesis remain unknown. The objective of this study was to elucidate the direct effects of cigarette smoke on human bone marrow hematopoiesis and characterize the inflammatory process known to result from cigarette smoking. Bone marrow mononuclear cells (BMCs from healthy individuals when exposed to CSE had significantly diminished CFU-E, BFU-E and CFU-GM. We found increased nuclear translocation of the NF-κB p65 subunit and, independently, enhanced activation of AKT and ERK1/2. Exposure of BMCs to CSE induced IL-8 and TGF-β1 production, which was dependent on NF-κB and ERK1/2, but not on AKT. CSE treatment had no effect on the release of TNF-α, IL-10, or VEGF. Finally, CSE also had a significant induction of TLR2, TLR3 and TLR4, out of which, the up-regulation of TLR2 and TLR3 was found to be dependent on ERK1/2 and NF-κB activation, but not AKT. These results indicate that CSE profoundly inhibits the growth of erythroid and granulocyte-macrophage progenitors in the bone marrow. Further, CSE modulates NF-κB- and ERK1/2-dependent responses, suggesting that cigarette smoking may impair bone marrow hematopoiesis in vivo as well as induce inflammation, two processes that proceed malignant transformation.

  7. PPInterFinder--a mining tool for extracting causal relations on human proteins from literature.

    Science.gov (United States)

    Raja, Kalpana; Subramani, Suresh; Natarajan, Jeyakumar

    2013-01-01

    One of the most common and challenging problem in biomedical text mining is to mine protein-protein interactions (PPIs) from MEDLINE abstracts and full-text research articles because PPIs play a major role in understanding the various biological processes and the impact of proteins in diseases. We implemented, PPInterFinder--a web-based text mining tool to extract human PPIs from biomedical literature. PPInterFinder uses relation keyword co-occurrences with protein names to extract information on PPIs from MEDLINE abstracts and consists of three phases. First, it identifies the relation keyword using a parser with Tregex and a relation keyword dictionary. Next, it automatically identifies the candidate PPI pairs with a set of rules related to PPI recognition. Finally, it extracts the relations by matching the sentence with a set of 11 specific patterns based on the syntactic nature of PPI pair. We find that PPInterFinder is capable of predicting PPIs with the accuracy of 66.05% on AIMED corpus and outperforms most of the existing systems. DATABASE URL: http://www.biomining-bu.in/ppinterfinder/ PMID:23325628

  8. The Effect of Cinnamon Extract on Serum Proteins Levels of Male Balb/c Mice

    OpenAIRE

    M Modaresi

    2011-01-01

    Background & Aim: Cinnamon is a plant with many pharmaceutical effects. The present research evaluated the effects of Cinnamon bark extract on serum proteins level in male Balb/c mice. Methods: In this experimental study, 40 small Balb/c mice were chosen and divided into 5 groups: a control group, a case group, and three treatment groups. Normal saline was administered as placebo to the case group while the control group received no injections.. Cinnamon extract in doses of 50, 100 and 20...

  9. Sunflower seed protein concentrates and isolates obtention from ethanol oil extraction meals--(technical note).

    Science.gov (United States)

    Regitano-d'Arce, M A; Gutierrez, E M; Lima, U de A

    1994-03-01

    The objective of this work was to study and identify the necessary processing steps for obtaining good quality sunflower seed protein concentrate and isolate when the oil is extracted with ethanol. This work is part of a research project on using ethanol as renewable solvent for sunflower seed oil recovery and possible further processing of the meal. Both 99 degrees GL and 90 degrees GL ethanol were employed in the extractions to produce the concentrate. Isolates were obtained by treating the concentrate with NaOH and HCl solutions and final rinsing with acidified water. Both products were light in color and almost free from chlorogenic acid. PMID:7717804

  10. Iron carrier proteins facilitate engraftment of allogeneic bone marrow and enduring hemopoietic chimerism in the lethally irradiated host

    International Nuclear Information System (INIS)

    Cell-free supernatants of rabbit bone marrow were fractionated, separated, and purified by Ultrogel and Superose chromatography. A single fraction promoted engraftment of allogeneic bone marrow and enduring hemopoietic chimerism across the H-2 barrier in lethally irradiated mice. This 'bio-active' fraction, analyzed by reducing SDS-PAGE electrophoresis, and transblotted on PVDF membrane, and purified by reverse-phase HPLC and SDS-PAGE electrophoresis yielded a main prealbumin band that when examined for primary structure by Edman degradation, proved to be rabbit transferrin. This was also attested by highly specific precipitation of the prealbumin band with polyclonal antibodies to rabbit transferrin. Iron-saturated human transferrin, lactotransferrin, and egg transferrin (conalbumin) were assayed in irradiated C57BL/6 mice infused with bone marrow from histoincompatible BALB/c donors. Mice treated with iron-loaded transferrins survive and develop enduring allogeneic chimerism with no discernible signs of graft-versus-host disease. Iron carrier proteins thus provide an unique means of achieving successful engraftment of allogeneic bone marrow in immunologically hostile murine H-2 combinations

  11. Bone morphogenetic protein-9 suppresses growth of myeloma cells by signaling through ALK2 but is inhibited by endoglin

    International Nuclear Information System (INIS)

    Multiple myeloma is a malignancy of plasma cells predominantly located in the bone marrow. A number of bone morphogenetic proteins (BMPs) induce apoptosis in myeloma cells in vitro, and with this study we add BMP-9 to the list. BMP-9 has been found in human serum at concentrations that inhibit cancer cell growth in vitro. We here show that the level of BMP-9 in serum was elevated in myeloma patients (median 176 pg/ml, range 8–809) compared with healthy controls (median 110 pg/ml, range 8–359). BMP-9 was also present in the bone marrow and was able to induce apoptosis in 4 out of 11 primary myeloma cell samples by signaling through ALK2. BMP-9-induced apoptosis in myeloma cells was associated with c-MYC downregulation. The effects of BMP-9 were counteracted by membrane-bound (CD105) or soluble endoglin present in the bone marrow microenvironment, suggesting a mechanism for how myeloma cells can evade the tumor suppressing activity of BMP-9 in multiple myeloma

  12. Sustained and promoter dependent bone morphogenetic protein expression by rat mesenchymal stem cells after BMP-2 transgene electrotransfer

    Directory of Open Access Journals (Sweden)

    E Ferreira

    2012-07-01

    Full Text Available Transplantation of mesenchymal stem cells (MSCs with electrotransferred bone morphogenetic protein-2 (BMP-2 transgene is an attractive therapeutic modality for the treatment of large bone defects: it provides both stem cells with the ability to form bone and an effective bone inducer while avoiding viral gene transfer. The objective of the present study was to determine the influence of the promoter driving the human BMP-2 gene on the level and duration of BMP-2 expression after transgene electrotransfer into rat MSCs. Cytomegalovirus, elongation factor-1α, glyceraldehyde 3-phosphate dehydrogenase, and beta-actin promoters resulted in a BMP-2 secretion rate increase of 11-, 78-, 66- and 36-fold over respective controls, respectively. In contrast, the osteocalcin promoter had predictable weak activity in undifferentiated MSCs but induced the strongest BMP-2 secretion rates in osteoblastically-differentiated MSCs. Regardless of the promoter driving the transgene, a plateau of maximal BMP-2 secretion persisted for at least 21 d after the hBMP-2 gene electrotransfer. The present study demonstrates the feasibility of gene electrotransfer for efficient BMP-2 transgene delivery into MSCs and for a three-week sustained BMP-2 expression. It also provides the first in vitro evidence for a safe alternative to viral methods that permit efficient BMP-2 gene delivery and expression in MSCs but raise safety concerns that are critical when considering clinical applications.

  13. 5-Azacytidine-induced protein 2 (AZI2) regulates bone mass by fine-tuning osteoclast survival.

    Science.gov (United States)

    Maruyama, Kenta; Fukasaka, Masahiro; Uematsu, Satoshi; Takeuchi, Osamu; Kondo, Takeshi; Saitoh, Tatsuya; Martino, Mikaël M; Akira, Shizuo

    2015-04-10

    5-Azacytidine-induced protein 2 (AZI2) is a TNF receptor (TNFR)-associated factor family member-associated NF-κB activator-binding kinase 1-binding protein that regulates the production of IFNs. A previous in vitro study showed that AZI2 is involved in dendritic cell differentiation. However, the roles of AZI2 in immunity and its pleiotropic functions are unknown in vivo. Here we report that AZI2 knock-out mice exhibit normal dendritic cell differentiation in vivo. However, we found that adult AZI2 knock-out mice have severe osteoporosis due to increased osteoclast longevity. We revealed that the higher longevity of AZI2-deficient osteoclasts is due to an augmented activation of proto-oncogene tyrosine-protein kinase Src (c-Src), which is a critical player in osteoclast survival. We found that AZI2 inhibits c-Src activity by regulating the activation of heat shock protein 90 (Hsp90), a chaperone involved in c-Src dephosphorylation. Furthermore, we demonstrated that AZI2 indirectly inhibits c-Src by interacting with the Hsp90 co-chaperone Cdc37. Strikingly, administration of a c-Src inhibitor markedly prevented bone loss in AZI2 knock-out mice. Together, these findings indicate that AZI2 regulates bone mass by fine-tuning osteoclast survival. PMID:25691576

  14. 5-Azacytidine-induced Protein 2 (AZI2) Regulates Bone Mass by Fine-tuning Osteoclast Survival*

    Science.gov (United States)

    Maruyama, Kenta; Fukasaka, Masahiro; Uematsu, Satoshi; Takeuchi, Osamu; Kondo, Takeshi; Saitoh, Tatsuya; Martino, Mikaël M.; Akira, Shizuo

    2015-01-01

    5-Azacytidine-induced protein 2 (AZI2) is a TNF receptor (TNFR)-associated factor family member-associated NF-κB activator-binding kinase 1-binding protein that regulates the production of IFNs. A previous in vitro study showed that AZI2 is involved in dendritic cell differentiation. However, the roles of AZI2 in immunity and its pleiotropic functions are unknown in vivo. Here we report that AZI2 knock-out mice exhibit normal dendritic cell differentiation in vivo. However, we found that adult AZI2 knock-out mice have severe osteoporosis due to increased osteoclast longevity. We revealed that the higher longevity of AZI2-deficient osteoclasts is due to an augmented activation of proto-oncogene tyrosine-protein kinase Src (c-Src), which is a critical player in osteoclast survival. We found that AZI2 inhibits c-Src activity by regulating the activation of heat shock protein 90 (Hsp90), a chaperone involved in c-Src dephosphorylation. Furthermore, we demonstrated that AZI2 indirectly inhibits c-Src by interacting with the Hsp90 co-chaperone Cdc37. Strikingly, administration of a c-Src inhibitor markedly prevented bone loss in AZI2 knock-out mice. Together, these findings indicate that AZI2 regulates bone mass by fine-tuning osteoclast survival. PMID:25691576

  15. Mandibular defect reconstruction using three-dimensional polycaprolactone scaffold in combination with platelet-rich plasma and recombinant human bone morphogenetic protein-2: de novo synthesis of bone in a single case.

    Science.gov (United States)

    Schuckert, Karl-Heinz; Jopp, Stefan; Teoh, Swee-Hin

    2009-03-01

    This publication describes the clinical case of a 71-year-old female patient. Using polycaprolactone (PCL) scaffold, platelet-rich plasma (PRP) and recombinant human bone morphogenetic protein-2 (rhBMP-2), a critical-sized defect in the anterior mandible was regenerated using de novo-grown bone. A bacterial infection had caused a periimplantitis in two dental implants leading to a large destruction in the anterior mandible. Both implants were removed under antibiotic prophylaxis. A PCL scaffold was prepared especially for this clinical case. In a second procedure with antibiotic prophylaxis, the bony defect was reopened. The PCL scaffold was fitted and charged with PRP and rhBMP-2 (1.2 mg). After complication-free wound healing, the radiological control demonstrated de novo-grown bone in the anterior mandible 6 months postoperatively. Dental implants were inserted in a third operation. A bone biopsy of the newly grown bone, as well as of the bordering local bone, was taken and histologically examined. The bone samples were identical and presented vital laminar bone. PMID:18767969

  16. Aqueous two-phase system based on natural quaternary ammonium compounds for the extraction of proteins.

    Science.gov (United States)

    Zeng, Chao-Xi; Xin, Rui-Pu; Qi, Sui-Jian; Yang, Bo; Wang, Yong-Hua

    2016-02-01

    Aqueous two-phase systems, based on the use of natural quaternary ammonium compounds, were developed to establish a benign biotechnological route for efficient protein separation. In this study, aqueous two-phase systems of two natural resources betaine and choline with polyethyleneglycol (PEG400/600) or inorganic salts (K2 HPO4 /K3 PO4 ) were formed. It was shown that in the K2 HPO4 -containing aqueous two-phase system, hydrophobic interactions were an important driving force of protein partitioning, while protein size played a vital role in aqueous two-phase systems that contained polyethylene glycol. An extraction efficiency of more than 90% for bovine serum albumin in the betaine/K2 HPO4 aqueous two-phase system can be obtained, and this betaine-based aqueous two-phase system provided a gentle and stable environment for the protein. In addition, after investigation of the cluster phenomenon in the betaine/K2 HPO4 aqueous two-phase systems, it was suggested that this phenomenon also played a significant role for protein extraction in this system. The development of aqueous two-phase systems based on natural quaternary ammonium compounds not only provided an effective and greener method of aqueous two-phase system to meet the requirements of green chemistry but also may help to solve the mystery of the compartmentalization of biomolecules in cells. PMID:26447826

  17. Effect of extraction method on functional properties of flaxseed protein concentrates.

    Science.gov (United States)

    Tirgar, Mina; Silcock, Patrick; Carne, Alan; Birch, E John

    2017-01-15

    Physicochemical (zeta potential (ζ), conductivity, surface hydrophobicity (H0), protein solubility (PS)) and emulsifying (emulsion capacity (EC), droplet size, polydispersity (PDI), emulsifying activity (EAI), and stability (ESI) indexes) properties of alkali-(A-FPC), enzymatic-(E-FPC), and enzymatic-solvent-(ES-FPC) extracted protein concentrates from flaxseed meal (FM) were investigated and compared to commercial pea protein concentrate (PPC). The yield, composition, and properties of the protein concentrates were significantly influenced by the methods of extraction. All emulsions were similar in polydispersity with mono-modal droplet distribution and size of ⩽0.43μm that carried a net negative charge at neutral conditions (pH 7.0). A-FPC showed significantly higher H0 (66.14) than that of ES-FPC (52.63), and E-FPC (43.27) and was comparable to PPC (68.47). The highest solubility was found for E-FPC followed by A-FPC at neutral pH. A-FPC displayed significantly (p<0.05) the highest EC (87.91%), EAI (87.18m(2)/g) and ESI (12.51min) compared to the other protein concentrates. PMID:27542494

  18. Candida albicans PROTEIN PROFILE CHANGES IN RESPONSE TO THE BUTANOLIC EXTRACT OF Sapindus saponariaL.

    Directory of Open Access Journals (Sweden)

    Adriana FIORINI

    2016-01-01

    Full Text Available Candida albicans is an opportunistic human pathogen that is capable of causing superficial and systemic infections in immunocompromised patients. Extracts of Sapindus saponaria have been used as antimicrobial agents against various organisms. In the present study, we used a combination of two-dimensional polyacrylamide gel electrophoresis (2D-PAGE and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS to identify the changes in protein abundance of C. albicans after exposure to the minimal inhibitory concentration (MIC and sub-minimal inhibitory concentration (sub-MIC of the butanolic extract (BUTE of S. saponaria and also to fluconazole. A total of six different proteins with greater than 1.5 fold induction or repression relative to the untreated control cells were identified among the three treatments. In general, proteins/enzymes involved with the glycolysis (GPM1, ENO1, FBA1, amino acid metabolism (ILV5, PDC11 and protein synthesis (ASC1 pathways were detected. In conclusion, our findings reveal antifungal-induced changes in protein abundance of C. albicans. By using the previously identified components of the BUTE of S. saponaria(e.g., saponins and sesquiterpene oligoglycosides, it will be possible to compare the behavior of compounds with unknown mechanisms of action, and this knowledge will help to focus the subsequent biochemical work aimed at defining the effects of these compounds.

  19. Candida albicans PROTEIN PROFILE CHANGES IN RESPONSE TO THE BUTANOLIC EXTRACT OF Sapindus saponariaL.

    Science.gov (United States)

    FIORINI, Adriana; ROSADO, Fabio Rogério; BETTEGA, Eliane Martins da Silva; MELO, Kátia Cristina Sibin; KUKOLJ, Caroline; BONFIM-MENDONÇA, Patrícia de Souza; SHINOBU-MESQUITA, Cristiane Suemi; GHIRALDI, Luciana Dias; CAMPANERUT, Paula Aline Zanetti; CAPOCI, Isis Regina Grenier; GODOY, Janine Silva Ribeiro; FERREIRA, Izabel Cristina Piloto; SVIDZINSKI, Terezinha Inez Estivalet

    2016-01-01

    Candida albicans is an opportunistic human pathogen that is capable of causing superficial and systemic infections in immunocompromised patients. Extracts of Sapindus saponaria have been used as antimicrobial agents against various organisms. In the present study, we used a combination of two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to identify the changes in protein abundance of C. albicans after exposure to the minimal inhibitory concentration (MIC) and sub-minimal inhibitory concentration (sub-MIC) of the butanolic extract (BUTE) of S. saponaria and also to fluconazole. A total of six different proteins with greater than 1.5 fold induction or repression relative to the untreated control cells were identified among the three treatments. In general, proteins/enzymes involved with the glycolysis (GPM1, ENO1, FBA1), amino acid metabolism (ILV5, PDC11) and protein synthesis (ASC1) pathways were detected. In conclusion, our findings reveal antifungal-induced changes in protein abundance of C. albicans. By using the previously identified components of the BUTE of S. saponaria(e.g., saponins and sesquiterpene oligoglycosides), it will be possible to compare the behavior of compounds with unknown mechanisms of action, and this knowledge will help to focus the subsequent biochemical work aimed at defining the effects of these compounds. PMID:27074319

  20. Increased expression of osteonectin and osteopontin, two bone matrix proteins, in human breast cancer.

    OpenAIRE

    Bellahcène, A.; Castronovo, V.

    1995-01-01

    Microcalcifications are a common phenomenon associated with breast cancer and are often the only mammographic sign of a malignant breast disease. Although microcalcifications are not restricted to breast cancer and can be also associated with benign lesions, it is noteworthy that they are composed exclusively of hydroxyapatite in breast carcinoma. Hydroxyapatite is the bone-associated phosphocalcic crystal the deposition of which in bone tissue requires the coordinated expression of several m...

  1. Using co-occurrence network structure to extract synonymous gene and protein names from MEDLINE abstracts

    Directory of Open Access Journals (Sweden)

    Spackman K

    2005-04-01

    Full Text Available Abstract Background Text-mining can assist biomedical researchers in reducing information overload by extracting useful knowledge from large collections of text. We developed a novel text-mining method based on analyzing the network structure created by symbol co-occurrences as a way to extend the capabilities of knowledge extraction. The method was applied to the task of automatic gene and protein name synonym extraction. Results Performance was measured on a test set consisting of about 50,000 abstracts from one year of MEDLINE. Synonyms retrieved from curated genomics databases were used as a gold standard. The system obtained a maximum F-score of 22.21% (23.18% precision and 21.36% recall, with high efficiency in the use of seed pairs. Conclusion The method performs comparably with other studied methods, does not rely on sophisticated named-entity recognition, and requires little initial seed knowledge.

  2. Effect of nigella sativa seeds extract on serum c-reactive protein in albino rats

    International Nuclear Information System (INIS)

    C-reactive protein (CRP) is an acute phase protein. It predicts future risk of cardiovascular diseases. Different medicinal plants and their active ingredients possess the ability to reduce serum CRP levels and hence inflammatory disorders and cardiovascular diseases. In our study, ethanolic extract of Nigella sativa seeds was evaluated in albino rats for its possible effect on serum CRP levels. Objective: The objective of this study was to determine the effect of ethanolic extract of Nigella sativa seeds on an acute inflammatory biomarker/mediator, C-reactive protein (CRP) in albino rats. Study Design: Randomized controlled trial (RCT). Place and Duration of Study: Physiology Department, Services Institute of Medical Sciences (SIMS), Lahore; from September to November, 2009. Subjects and Methods: The study was carried out on 90 male albino rats. Five percent (5%) formalin in a dose of 50 meu1 was injected into sub-plantar surface of right hind paw of each rat to produce inflammation. The rats were randomly divided into three groups of thirty each. Group A was given normal saline (control); group B was given Nigella sativa seed extract; and group C received diclofenac sodium, as a reference drug. CRP levels in each group were measured from blood samples taken 25 hours after giving formalin. Results: The ethanolic extract of Nigella sativa seeds, given intraperitoneally, caused highly significant (p<0.001) reduction in serum CRP levels as compared to control group. The reduction in CRP levels by ethanolic extract of Nigella sativa was also significantly (p<0.05) more than that produced by diclofenac sodium. Conclusion: Our results suggest that Nigella sativa possesses ability to reduce serum CRP levels significantly, after production of artificial inflammation, in albino rats. (author)

  3. Total Protein Extraction for Metaproteomics Analysis of Methane Producing Biofilm: The Effects of Detergents

    OpenAIRE

    Hung-Jen Huang; Wei-Yu Chen; Jer-Horng Wu

    2014-01-01

    Protein recovery is crucial for shotgun metaproteomics to study the in situ functionality of microbial populations from complex biofilms but still poorly addressed by far. To fill this knowledge gap, we systematically evaluated the sample preparation with extraction buffers comprising four detergents for the metaproteomics analysis of a terephthalate-degrading methanogenic biofilm using an on-line two-dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS) system. Totally...

  4. Azorella compacta methanolic extract induces apoptosis via activation of mitogen-activated protein kinase.

    Science.gov (United States)

    Sung, Min Hee; Kwon, Ok-Kyoung; Oh, Sei-Ryang; Lee, Joongku; Park, Sang-Hong; Han, Sang Bae; Ahn, Kyung-Seop

    2015-11-01

    Azorella compacta Phil. (AC) is an alpine medicinal plant used traditionally for antibacterial treatment. Recent studies have revealed that this plant also has anti‑diabetic effects, but that it is toxic. The present study investigated the underlying mechanisms of action of AC extract against human leukemia HL60 cells. Apoptosis induction was measured by MTT assay, fluorescence microscopy, DNA fragmentation assay, flow cytometric analysis, reverse transcription quantitative polymerase chain reaction and western blot analyses. It was found that AC extract inhibited the growth of HL60 and other cancer cell lines in a dose‑dependent manner. The cytotoxic effects of AC extract on HL60 cells were associated with apoptosis characterized by DNA fragmentation and dose‑dependent increases in Annexin V‑positive cells, as determined by flow cytometric analysis. AC‑extract‑induced apoptosis was accompanied by activated/cleaved caspase‑3, caspase‑9 and poly(adenosine diphosphate‑ribose) polymerase (PARP). The increases in apoptosis were also associated with decreases of the apoptosis-inhibitor B-cell lymphoma 2 (Bcl‑2), upregulation of pro‑apoptotic Bcl-2-associated X (Bax) protein and downregulation of anti‑apoptotic Bcl extra large protein. Furthermore, western blot analysis of mitogen-activated protein kinase (MAPK)-associated proteins indicated that treatment with AC extract increased the levels of c-Jun N-terminal kinase, extracellular signal-regulated kinase and p38. In addition, the expression of Bax and cleaved PARP was blocked when AC treatment was performed in the presence of MAPK inhibitors. It was therefore concluded that AC induced apoptosis in human leukemia HL60 cells via an intrinsic pathway controlled through MAPK-associated signaling. PMID:26397193

  5. Antiatherosclerotic Effect of Korean Red Ginseng Extract Involves Regulator of G-Protein Signaling 5

    OpenAIRE

    2014-01-01

    Regulator of G-protein signaling 5 (RGS5), an inhibitor of Gα(q) and Gα(i) activation, has been reported to have antiatherosclerosis. Previous studies showed antiatherosclerotic effect of Korean red ginseng water extract (KRGE) via multiple signaling pathways. However, potential protective effect of KRGE through RGS5 expression has not been elucidated. Here, we investigated the antiatherosclerotic effect of KRGE in vivo and in vitro and its role on RGS5 mRNA expression. Elevated levels of tot...

  6. Isolation of proteins from ejaculated and capacitated sperms by various extraction approaches

    Czech Academy of Sciences Publication Activity Database

    Zigo, Michal; Jonáková, Věra; Postlerová, Pavla

    2010-01-01

    Roč. 33, Supplement 1 (2010), s. 68-68. ISSN 0105-6263. [6th European Congress of And rology. 29.09.2010-01.10.2010, Athens] R&D Projects: GA ČR(CZ) GA303/09/1285; GA MŠk(CZ) 1M06011; GA MZd NS10009-4/2008 Institutional research plan: CEZ:AV0Z50520701 Keywords : spermatozoa * protein extraction * 2D electrophoresis * zona pellucida * glycoproteins Subject RIV: CE - Biochemistry

  7. The Use of Bone Morphogenetic Protein in Pediatric Cervical Spine Fusion Surgery: Case Reports and Review of the Literature.

    Science.gov (United States)

    Molinari, Robert W; Molinari, Christine

    2016-02-01

    Study Design Case report. Objective There is a paucity of literature describing the use of bone graft substitutes to achieve fusion in the pediatric cervical spine. The outcomes and complications involving the off-label use of bone morphogenetic protein (BMP)-2 in the pediatric cervical spine are not clearly defined. The purpose of this article is to report successful fusion without complications in two pediatric patients who had instrumented occipitocervical fusion using low-dose BMP-2. Methods A retrospective review of the medical records was performed, and the patients were followed for 5 years. Two patients under 10 years of age with upper cervical instability were treated with occipitocervical instrumented fusion using rigid occipitocervical fixation techniques along with conventionally available low-dose BMP-2. A Medline and PubMed literature search was conducted using the terms "bone morphogenetic protein," "BMP," "rh-BMP2," "bone graft substitutes," and "pediatric cervical spine." Results Solid occipitocervical fusion was achieved in both pediatric patients. There were no reported perioperative or follow-up complications. At 5-year follow-up, radiographs in both patients showed successful occipital cervical fusion without evidence of instrumentation failure or changes in the occipitocervical alignment. To date, there are few published reports on this topic. Complications and the appropriate dosage application in the pediatric posterior cervical spine remain unknown. Conclusions We describe two pediatric patients with upper cervical instability who achieved successful occipital cervical fusion without complication using off-label BMP-2. This report underscores the potential for BMP-2 to achieve successful arthrodesis of the posterior occipitocervical junction in pediatric patients. Use should be judicious as complications and long-term outcomes of pediatric BMP-2 use remain undefined in the existing literature. PMID:26835215

  8. Optimization of Protein Extraction and Two-Dimensional Electrophoresis Protocols for Oil Palm Leaf.

    Science.gov (United States)

    Daim, Leona Daniela Jeffery; Ooi, Tony Eng Keong; Yusof, Hirzun Mohd; Majid, Nazia Abdul; Karsani, Saiful Anuar Bin

    2015-08-01

    Oil palm (Elaeis guineensis) is an important economic crop cultivated for its nutritional palm oil. A significant amount of effort has been undertaken to understand oil palm growth and physiology at the molecular level, particularly in genomics and transcriptomics. Recently, proteomics studies have begun to garner interest. However, this effort is impeded by technical challenges. Plant sample preparation for proteomics analysis is plagued with technical challenges due to the presence of polysaccharides, secondary metabolites and other interfering compounds. Although protein extraction methods for plant tissues exist, none work universally on all sample types. Therefore, this study aims to compare and optimize different protein extraction protocols for use with two-dimensional gel electrophoresis of young and mature leaves from the oil palm. Four protein extraction methods were evaluated: phenol-guanidine isothiocyanate, trichloroacetic acid-acetone precipitation, sucrose and trichloroacetic acid-acetone-phenol. Of these four protocols, the trichloroacetic acid-acetone-phenol method was found to give the highest resolution and most reproducible gel. The results from this study can be used in sample preparations of oil palm tissue for proteomics work. PMID:26263918

  9. Two-dimensional electrophoresis analysis of proteins extracted from Alexandrium sp. LC3

    Science.gov (United States)

    Li, Hao; Miao, Jinlai; Cui, Fengxia; Li, Guangyou

    2007-10-01

    Two-dimensional electrophoresis(2-DE) of protein extracted and purified from Alexandrium sp. LC3 was conducted. In the SDS-PAGE study, the relative molecular weights of the proteins were mainly in the range of 14kDa-31kDa and 43kDa-66kDa, and more proteins were detected between 14kDa and 31kDa. With the improved protein preparation, the two-dimensional electrophoresis patterns indicated that the relative molecular weights of the proteins were between 14kDa and 100kDa, and most of them ranged from 14kDa to 31kDa. This was consistent with the result of the SDS-PAGE analysis. The isoelectric points were found to lie between 3.0 and 8.0, and most of them were in the range of 3.0 6.0. Better separation effect was acquired with pre-prepared immobilized gradient (IPG) strip (pH3 5.6), and about 320 protein spots could be visualized on the 2-DE map by staining. Within pH3 10 and pH3 5.6 strips, the protein samples of Alexandrium sp. LC3 could be separated well.

  10. Salting-in effect on muscle protein extracted from giant squid (Dosidicus gigas).

    Science.gov (United States)

    Zhang, Rui; Zhou, Ru; Pan, Weichun; Lin, Weiwei; Zhang, Xiuzhen; Li, Mengya; Li, Jianrong; Niu, Fuge; Li, Ang

    2017-01-15

    The salting-in effect on muscle protein is well-known in food science but hard to explain using conventional theories. Myofibrillar protein extracted from the giant squid (Dosidicus gigas) was selected as a model muscle protein to study this mechanism in KCl solutions. Changes in the secondary structures of myofibrillar protein molecules caused by concentrated salts, particularly in the paramyosin molecule conformation, have been reported. Zeta-potential determinations showed that these secondary structures have modified protein molecule surfaces. The zeta-potential of the myofibrillar protein molecules fell from -7.24±0.82 to -9.99±1.65mV with increasing salt concentration from 0.1 to 0.5M. The corresponding second virial coefficient increased from -85.43±3.8×10(-7) to -3.45±1.3×10(-7) molmLg(-2). The extended law of corresponding states suggests that reduced attractive interactions increase the protein solubility. Solubility measurements in alternating KCl concentrations showed that the conformational change was reversible. PMID:27542474

  11. Environmental pH-controlled loading and release of protein on mesoporous hydroxyapatite nanoparticles for bone tissue engineering.

    Science.gov (United States)

    Zhang, Ning; Gao, Tianlin; Wang, Yu; Wang, Zongliang; Zhang, Peibiao; Liu, Jianguo

    2015-01-01

    To explore the controlled delivery of protein drugs in micro-environment established by osteoblasts or osteoclasts, the loading/release properties of bovine serum albumin (BSA) depending on pH environment were assessed. The adsorption amounts over mesoporous hydroxyapatite (MHA) or hydroxyapatite (HA) decreased as the pH increased, negatively correlating with zeta-potential values. The adsorption behavior over MHA fits well with the Freundlich and Langmuir models at different pHs. The results suggest that the adsorbed amount of protein on MHA or HA depended on the pH of protein solution. MHA adsorbed BSA at basic pH (MHApH 8.4) exhibited a different release kinetics compared with those in acid and neutral environments (MHApH 4.7 and MHApH 7.4), indicating that the release of protein could be regulated by environmental pH at which MHAs adsorb protein. MHApH 8.4 showed a sustained release for 6h before a gradual release when immersing in acidic environment, which is 2h longer than that in neutral environment. This suggests that MHApH 8.4 showed a more sustained release in acidic environment, which can be established by osteoclasts. The variation of adsorption strength between protein and MHA may be responsible for these behaviors. Our findings may be very useful for the development of MHA applications on both bone repair and protein delivery. PMID:25491972

  12. Validation of a Non-Targeted LC-MS Approach for Identifying Ancient Proteins: Method Development on Bone to Improve Artifact Residue Analysis

    OpenAIRE

    Andrew Barker; Jonathan Dombrosky; Dale Chaput; Barney Venbles; Steve Wolverton; Stevens, Stanley M.

    2015-01-01

    Identification of protein residues from prehistoric cooking pottery using mass spectrometry is challenging because proteins are removed from original tissues, are degraded from cooking, may be poorly preserved due to diagenesis, and occur in a palimpsest of exogenous soil proteins. In contrast, bone proteins are abundant and well preserved. This research is part of a larger method-development project for innovation and improvement of liquid chromatography – mass spectrometry analysis of prote...

  13. Functional assay, expression of growth factors and proteins modulating bone-arrangement in human osteoblasts seeded on an anorganic bovine bone biomaterial

    OpenAIRE

    O Trubiani; Fulle, S.; T Traini; M Paludi; La Rovere, R.; M Orciani; S. Caputi; Piattelli, A.

    2010-01-01

    The basic aspects of bone tissue engineering include chemical composition and geometry of the scaffold design, because it is very important to improve not only cell attachment and growth but especially osteodifferentiation, bone tissue formation, and vascularization. Geistlich Bio-Oss® (GBO) is a xenograft consisting of deproteinized, sterilized bovine bone, chemically and physically identical to the mineral phase of human bone.In this study, we investigated the growth behaviour and the abili...

  14. Quantitative analysis of protein and gene expression in salivary glands of Sjogren's-like disease NOD mice treated by bone marrow soup.

    Directory of Open Access Journals (Sweden)

    Kaori Misuno

    Full Text Available BACKGROUND: Bone marrow cell extract (termed as BM Soup has been demonstrated to repair irradiated salivary glands (SGs and restore saliva secretion in our previous study. In the present study, we aim to investigate if the function of damaged SGs in non-obese diabetic (NOD mice can be restored by BM Soup treatment and the molecular alterations associated with the treatment. METHODS: Whole BM cells were lysed and soluble intracellular contents ("BM Soup" were injected I.V. into NOD mice. Tandem mass tagging with 2-D liquid chromatography-mass spectrometry was used to quantify proteins in the submandibular glands (SMGs between untreated and BM Soup-treated mice. Quantitative PCR was used to identify genes with altered expression in the treated mice. RESULTS BM SOUP: restored salivary flow rates to normal levels and significantly reduced the focus scores of SMGs in NOD mice. More than 1800 proteins in SMG cells were quantified by the proteomic approach. Many SMG proteins involved in inflammation and apoptosis were found to be down-regulated whereas those involved in salivary gland biology and development/regeneration were up-regulated in the BM Soup-treated mice. qPCR analysis also revealed expression changes of growth factors and cytokines in the SMGs of the treated NOD mice. CONCLUSION: BM Soup treatment is effective to restore the function of damaged SGs in NOD mice. Through gene/protein expression analysis, we have found that BM Soup treatment might effectuate via inhibiting apoptosis, focal adhesion and inflammation whereas promoting development, regeneration and differentiation of the SG cells in NOD mice. These findings provide important insights on the potential mechanisms underlying the BM Soup treatment for functional restoration of damaged SGs in NOD mice. Additional studies are needed to further confirm the identified target genes and their related signaling pathways that are responsible for the BM Soup treatment.

  15. Modulation of function of multidrug resistance associated-proteins by Kaempferia parviflora extracts and their components.

    Science.gov (United States)

    Patanasethanont, Denpong; Nagai, Junya; Matsuura, Chie; Fukui, Kyoko; Sutthanut, Khaetthareeya; Sripanidkulchai, Bung-orn; Yumoto, Ryoko; Takano, Mikihisa

    2007-07-01

    In this study, the effects of extracts and flavone derivatives from the rhizome of Kaempferia parviflora on multidrug resistance associated-proteins (MRP)-mediated transport in A549 cells were examined. The cells employed express MRP1 and MRP2, but not P-glycoprotein. The cellular accumulation of calcein, an MRP substrate, was significantly increased by various MRP inhibitors without being affected by verapamil, a typical P-glycoprotein inhibitor. Ethanol and aqueous extracts from K. parviflora rhizome increased the accumulation of calcein and doxorubicin in A549 cells in a concentration-dependent manner. The inhibitory potency of the ethanol extract for MRP function was greater than that of the aqueous extract. Among six flavone derivatives isolated from K. parviflora rhizome, 5,7-dimethoxyflavone exhibited a maximal stimulatory effect on the accumulation of doxorubicin in A549 cells. The accumulation of doxorubicin was increased by four flavone derivatives without 5-hydroxy group, but not by the other two flavone derivatives with 5-hydroxy group. In addition, 5,7-dimethoxyflavone and 3,5,7,3',4'-pentamethoxyflavone decreased resistance to doxorubicin in A549 cells. These findings indicate that extracts and flavone derivatives from the rhizome of K. parviflora suppress MRP function, and therefore may be useful as modulators of multidrug resistance in cancer cells. PMID:17481606

  16. Protein Synthesis Inhibition Activity by Strawberry Tissue Protein Extracts during Plant Life Cycle and under Biotic and Abiotic Stresses

    Directory of Open Access Journals (Sweden)

    Walther Faedi

    2013-07-01

    Full Text Available Ribosome-inactivating proteins (RIPs, enzymes that are widely distributed in the plant kingdom, inhibit protein synthesis by depurinating rRNA and many other polynucleotidic substrates. Although RIPs show antiviral, antifungal, and insecticidal activities, their biological and physiological roles are not completely understood. Additionally, it has been described that RIP expression is augmented under stressful conditions. In this study, we evaluated protein synthesis inhibition activity in partially purified basic proteins (hereafter referred to as RIP activity from tissue extracts of Fragaria × ananassa (strawberry cultivars with low (Dora and high (Record tolerance to root pathogens and fructification stress. Association between the presence of RIP activity and the crop management (organic or integrated soil, growth stage (quiescence, flowering, and fructification, and exogenous stress (drought were investigated. RIP activity was found in every tissue tested (roots, rhizomes, leaves, buds, flowers, and fruits and under each tested condition. However, significant differences in RIP distribution were observed depending on the soil and growth stage, and an increase in RIP activity was found in the leaves of drought-stressed plants. These results suggest that RIP expression and activity could represent a response mechanism against biotic and abiotic stresses and could be a useful tool in selecting stress-resistant strawberry genotypes.

  17. Management of subtrochanteric femur fractures with internal fixation and recombinant human bone morphogenetic protein-7 in a patient with osteopetrosis: a case report

    Directory of Open Access Journals (Sweden)

    Golden Robert D

    2010-05-01

    Full Text Available Abstract Introduction Osteopetrosis is a group of conditions characterized by defects in the osteoclastic function of the bone resulting in defective bone resorption. Clinically, the condition is characterized by a dense, sclerotic, deformed bone which, despite an increased density observable by radiography, often results in an increased propensity to fracture and delayed union. Case Presentation We report the case of a 27-year-old Asian man presenting with bilateral subtrochanteric femur fractures. He had a displaced right subtrochanteric femur fracture after a low-energy fall, which was treated surgically. The second fracture that our patient endured was diagnosed as a stress fracture ten weeks later when he complained of pain in the contralateral left thigh. By that time, the right-sided fracture exhibited no radiographic evidence of healing, and when the left-sided stress fracture was being treated surgically, bone grafting with recombinant human bone morphogenetic protein-7 was also performed on the right side. Conclusion While there are no data supporting the use of bone morphogenic proteins in the management of delayed healing in patients with osteopetrosis, no other reliable osteoinductive grafting options are available to treat this condition. Both fractures in our patient healed, but based on the serial radiographic assessment it is uncertain to what degree the recombinant human bone morphogenetic protein-7 may have contributed to the successful outcome. It may have also contributed to the formation of heterotopic bone around the fracture site. Further investigation of the effectiveness and indications of bone morphogenic protein use for the management of delayed fracture healing in patients with osteopetrosis is warranted.