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991.
992.
Yu‐Sin Jang Alok Malaviya Joungmin Lee Jung Ae Im Sang Yup Lee Julia Lee Moon‐Ho Eom Jung‐Hee Cho Do Young Seung 《Biotechnology progress》2013,29(4):1083-1088
Butanol is considered as a superior biofuel, which is conventionally produced by clostridial acetone‐butanol‐ethanol (ABE) fermentation. Among ABE, only butanol and ethanol can be used as fuel alternatives. Coproduction of acetone thus causes lower yield of fuel alcohols. Thus, this study aimed at developing an improved Clostridium acetobutylicum strain possessing enhanced fuel alcohol production capability. For this, we previously developed a hyper ABE producing BKM19 strain was further engineered to convert acetone into isopropanol. The BKM19 strain was transformed with the plasmid pIPA100 containing the sadh (primary/secondary alcohol dehydrogenase) and hydG (putative electron transfer protein) genes from the Clostridium beijerinckii NRRL B593 cloned under the control of the thiolase promoter. The resulting BKM19 (pIPA100) strain produced 27.9 g/l isopropanol‐butanol‐ethanol (IBE) as a fuel alcohols with negligible amount of acetone (0.4 g/l) from 97.8 g/l glucose in lab‐scale (2 l) batch fermentation. Thus, this metabolically engineered strain was able to produce 99% of total solvent produced as fuel alcohols. The scalability and stability of BKM19 (pIPA100) were evaluated at 200 l pilot‐scale fermentation, which showed that the fuel alcohol yield could be improved to 0.37 g/g as compared to 0.29 g/g obtained at lab‐scale fermentation, while attaining a similar titer. To the best of our knowledge, this is the highest titer of IBE achieved and the first report on the large scale fermentation of C. acetobutylicum for IBE production. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1083–1088, 2013 相似文献
993.
Beata Kolesinska Dieter Seebach Faustin Kamena Peter Walde Markus Sauer Barbara Windschiegl Mira Meyer‐Ács Marc Vor der Brüggen Sebastian Giehring 《化学与生物多样性》2013,10(1):1-38
After a survey of the special role, which the amino acid proline plays in the chemistry of life, the cell‐penetrating properties of polycationic proline‐containing peptides are discussed, and the widely unknown discovery by the Giralt group (J. Am. Chem. Soc. 2002 , 124, 8876) is acknowledged, according to which fluorescein‐labeled tetradecaproline is slowly taken up by rat kidney cells (NRK‐49F). Here, we describe details of our previously mentioned (Chem. Biodiversity 2004 , 1, 1111) observation that a hexa‐β3‐Pro derivative penetrates fibroblast cells, and we present the results of an extensive investigation of oligo‐L ‐ and oligo‐D ‐α‐prolines, as well as of oligo‐β2h‐ and oligo‐β3h‐prolines without and with fluorescence labels ( 1 – 8 ; Fig. 1). Permeation through protein‐free phospholipid bilayers is detected with the nanoFAST biochip technology (Figs. 2–4). This methodology is applied for the first time for quantitative determination of translocation rates of cell‐penetrating peptides (CPPs) across lipid bilayers. Cell penetration is observed with mouse (3T3) and human foreskin fibroblasts (HFF; Figs. 5 and 6–8, resp.). The stabilities of oligoprolines in heparin‐stabilized human plasma increase with decreasing chain lengths (Figs. 9–11). Time‐ and solvent‐dependent CD spectra of most of the oligoprolines (Figs. 13 and 14) show changes that may be interpreted as arising from aggregation, and broadening of the NMR signals with time confirms this assumption. 相似文献
994.
M. Manfrini C. Di Bona A. Canella E. Lucarelli A. Pellati A. D'Agostino G. Barbanti‐Bròdano M. Tognon 《Journal of cellular physiology》2013,228(6):1229-1237
Bio‐engineered scaffolds used in orthopedic clinical applications induce different tissue responses after implantation. In this study, non‐stoichiometric Mg2+ ions and stoichiometric apatites, which are used in orthopedic surgery as bone substitutes, have been assayed in vitro with human adult mesenchymal stem cells (hMSC) to evaluate cytocompatibility and osteoconductivity. hMSCs from the bone marrow aspirates of orthopedic patients were isolated and analyzed by flow cytometry for the surface markers Stro1, CD29, CD44, CD71, CD73, CD90, CD105 (positive) and CD45, CD235 (negative). The hMSC were analyzed for self‐renewal capacity and for differentiation potential. The hMSC, which were grown on different biomaterials, were analyzed for (i) cytotoxicity by AlamarBlue metabolic assay, (ii) osteoconductivity by ELISA for activated focal adhesion kinase, (iii) cytoskeleton organization by fluorescence microscopy, and (iv) cell morphology which was investigated by scan electron microscopy (SEM). Results indicate that isolated cell populations agree with minimal criteria for defining hMSC cultures. Non‐stoichiometric Mg2+ and stoichiometric apatites, in granular form, represent a more favorable environment for mesenchymal stem cell adhesion and growth compared to the non‐stoichiometric Mg2+ apatite, in nano‐structured paste form. This study indicates that different forms of biomaterials modulate osteoconductivity and cellular growth by differential activation focal adhesion kinase. J. Cell. Physiol. 228: 1229–1237, 2013. © 2012 Wiley Periodicals, Inc. 相似文献
995.
Ashok K. Chaturvedi Susan T. Weintraub Jose L. Lopez‐Ribot Floyd L. Wormley Jr. 《Proteomics》2013,13(23-24):3429-3441
Cryptococcus neoformans, the main causative agent of cryptococcosis, is a fungal pathogen that causes life‐threatening meningoencephalitis in immunocompromised patients. To date, there is no vaccine or immunotherapy approved to treat cryptococcosis. Cell‐ and antibody‐mediated immune responses collaborate to mediate optimal protection against C. neoformans infections. Accordingly, we identified cryptococcal protein fractions capable of stimulating cell‐ and antibody‐mediated immune responses and determined their efficacy to elicit protection against cryptococcosis. Proteins were extracted from C. neoformans and fractionated based on molecular mass. The fractions were then evaluated by immunoblot analysis for reactivity to serum extracted from protectively immunized mice and in cytokine recall assays for their efficacy to induce pro‐inflammatory and Th1‐type cytokine responses associated with protection. MS analysis revealed a number of proteins with roles in stress response, signal transduction, carbohydrate metabolism, amino acid synthesis, and protein synthesis. Immunization with select protein fractions containing immunodominant antigens induced significantly prolonged survival against experimental pulmonary cryptococcosis. Our studies support using the combination of immunological and proteomic approaches to identify proteins that elicit antigen‐specific antibody and Th1‐type cytokine responses. The immunodominant antigens that were discovered represent attractive candidates for the development of novel subunit vaccines for treatment and/or prevention of cryptococcosis. 相似文献
996.
Shun‐Fu Chang Heng Jung Chen Kam‐Fai Lee Tseng‐Hsi Lin Ting‐Ying Huang Chu‐Shan Choe Li‐Tsen Lin Cheng‐Nan Chen 《Cellular microbiology》2013,15(10):1722-1734
Porphyromonas gingivalis is a major pathogen in the initiation and progression of periodontal disease, which is recognized as a common complication of diabetes. ICAM‐1 expression by human gingival fibroblasts (HGFs) is crucial for regulating local inflammatory responses in inflamed periodontal tissues. However, the effect of P. gingivalis in a high‐glucose situation in regulating HGF function is not understood. The P. gingivalis strain CCUG25226 was used to study the mechanisms underlying the modulation of HGF ICAM‐1 expression by invasion of high‐glucose‐treated P. gingivalis (HGPg). A high‐glucose condition upregulated fimA mRNA expression in P. gingivalis and increased its invasion ability in HGFs. HGF invasion with HGPg induced increases in the expression of ICAM‐1. By using specific inhibitors and short hairpin RNA (shRNA), we have demonstrated that the activation of p38 MAPK and Akt pathways is critical for HGPg‐induced ICAM‐1 expression. Luciferase reporters and chromatin immunoprecipitation assays suggest that HGPg invasion increases NF‐κB‐ and Sp1‐DNA‐binding activities in HGFs. Inhibition of NF‐κB and Sp1 activations blocked the HGPg‐induced ICAM‐1 promoter activity and expression. The effect of HGPg on HGF signalling and ICAM‐1 expression is mediated by CXC chemokine receptor 4 (CXCR4). Our findings identify the molecular pathways underlying HGPg‐dependent ICAM‐1 expression in HGFs, providing insight into the effect of P. gingivalis invasion in HGFs. 相似文献
997.
Isabella V. Miller Graca Raposo Ulrich Welsch Olivia Prazeres da Costa Uwe Thiel Maria Lebar Martina Maurer Hans‐Ulrich Bender Irene von Luettichau Günther H. S. Richter Stefan Burdach Thomas G. P. Grunewald 《Biology of the cell / under the auspices of the European Cell Biology Organization》2013,105(7):289-303
998.
Accurate estimates of heritability () are necessary to assess adaptive responses of populations and evolution of fitness‐related traits in changing environments. For plants, estimates generally rely on maternal progeny designs, assuming that offspring are either half‐sibs or unrelated. However, plant mating systems often depart from half‐sib assumptions, this can bias estimates. Here, we investigate how to accurately estimate in nonmodel species through the analysis of sibling designs with a moderate genotyping effort. We performed simulations to investigate how microsatellite marker information available for only a subset of offspring can improve estimates based on maternal progeny designs in the presence of nonrandom mating, inbreeding in the parental population or maternal effects. We compared the basic family method, considering or not adjustments based on average relatedness coefficients, and methods based on the animal model. The animal model was used with average relatedness information, or with hybrid relatedness information: associating one‐generation pedigree and family assumptions, or associating one‐generation pedigree and average relatedness coefficients. Our results highlighted that methods using marker‐based relatedness coefficients performed as well as pedigree‐based methods in the presence of nonrandom mating (i.e. unequal male reproductive contributions, selfing), offering promising prospects to investigate in situ heritabilities in natural populations. In the presence of maternal effects, only the use of pairwise relatednesses through pedigree information improved the accuracy of estimates. In that case, the amount of father‐related offspring in the sibling design is the most critical. Overall, we showed that the method using both one‐generation pedigree and average relatedness coefficients was the most robust to various ecological scenarios. 相似文献
999.
STEPHANE A. P. DEROCLES MANUEL PLANTEGENEST JEAN‐YVES RASPLUS ALEXIA MARIE DARREN M. EVANS DAVID H. LUNT ANNE LE RALEC 《Systematic Entomology》2016,41(2):379-391
Aphidiinae are mostly composed of specialist parasitoids and the few species described as generalist are suspected to be composed of cryptic specialists, almost indistinguishable based on morphological characteristics. The use of molecular markers has proven to be a useful tool for revealing cryptic species complexes and here we use seven mitochondrial and nuclear gene fragments to study possible genetic differentiation among seven Aphidiinae generalists. Maximum likelihood (ML) trees and Bayesian Poisson tree processes (bPTP) models were conducted on each gene separately and on the seven genes together. The standard cytochrome c oxidase I barcode region appeared to be the most polymorphic and probably the best marker to reveal putative cryptic species. However, we showed with ML trees and bPTP models that a complementary use of mitochondrial and nuclear genes was the most relevant approach to reliably identify cryptic genetic clades in the Aphidiinae. Overall, most of the analysed generalist morphospecies were shown to be composed of subgroups related to the aphid host, some of them revealed as cryptic species by the species delimitation analysis. Further studies are needed to reveal the generality of this result in Aphidiinae. 相似文献
1000.
Fluorescence methods (VistaCam iX proof and DIAGNODent pen) for the detection of occlusal carious lesions in teeth recovered from archaeological context 下载免费PDF全文
Jacek Tomczyk Julian Komarnitki Marta Zalewska Tomasz Lekszycki Dorota Olczak‐Kowalczyk 《American journal of physical anthropology》2014,154(4):525-534
Diagnosis of occlusal enamel caries in archaeologically derived collections remains a controversial problem because the accumulation of contaminants in fissures can interfere with diagnosis. Certain novel light‐induced fluorescence methods, such as the DIAGNODent pen 2190 (DD) and VistaCam iX Proof (VC), have been used to detect dental caries in clinical settings. In this study, the abilities of DD and VC to detect initial enamel caries in archaeologically derived material is determined and compared with those of other methods (visual inspection, X‐ray, histology, and micro‐CT). Dental material encompassing the remains of 58 individuals, including a total of 380 teeth from each of three historical periods: modern Islamic (AD 1850–1950), Islamic (AD 600–1200) and late Roman (AD 200–400), obtained from two archaeological sites (Terqa and Tell Masaikh) located in the Middle Euphrates valley (Syria), were analyzed. VC was found to have excellent sensitivity (98), while DD obtained lower sensitivity (76) in detecting dental caries in its early stages. The results obtained by VC and micro‐CT, considered the most reliable imaging technique, were not statistically significant (P = 0.3068). By contrast, results obtained by DD and micro‐CT results, and DD and VC results were statistically significant (P < 0.0001, P = 0.0015, respectively). However the presence of dirt, stain, calculus, and plaque in the pits and fissures of the occlusal surface compromise correct diagnosis of caries by VC and DD. Consequently, for teeth recovered from archaeological contexts where staining, calculus and plaque are present, the best solution remains micro‐CT. Am J Phys Anthropol 154:525–534, 2014. © 2014 Wiley Periodicals, Inc. 相似文献