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61.
Elias DA Tollaksen SL Kennedy DW Mottaz HM Giometti CS McLean JS Hill EA Pinchuk GE Lipton MS Fredrickson JK Gorby YA 《Archives of microbiology》2008,189(4):313-324
High-throughput analyses that are central to microbial systems biology and ecophysiology research benefit from highly homogeneous
and physiologically well-defined cell cultures. While attention has focused on the technical variation associated with high-throughput
technologies, biological variation introduced as a function of cell cultivation methods has been largely overlooked. This
study evaluated the impact of cultivation methods, controlled batch or continuous culture in bioreactors versus shake flasks,
on the reproducibility of global proteome measurements in Shewanella
oneidensis MR-1. Variability in dissolved oxygen concentration and consumption rate, metabolite profiles, and proteome was greater in
shake flask than controlled batch or chemostat cultures. Proteins indicative of suboxic and anaerobic growth (e.g., fumarate
reductase and decaheme c-type cytochromes) were more abundant in cells from shake flasks compared to bioreactor cultures, a finding consistent with
data demonstrating that “aerobic” flask cultures were O2 deficient due to poor mass transfer kinetics. The work described herein establishes the necessity of controlled cultivation
for ensuring highly reproducible and homogenous microbial cultures. By decreasing cell to cell variability, higher quality
samples will allow for the interpretive accuracy necessary for drawing conclusions relevant to microbial systems biology research. 相似文献
62.
Up to 15% of all colorectal cancers are considered to be replication error positive (RER(+)) and contain mutations at hundreds of thousands of microsatellite repeat sequences. Recently, a number of intragenic mononucleotide repeat sequences have been demonstrated to be targets for inactivating genes in RER(+)colorectal tumors. In this study, thermostable DNA ligases were tested for the ability to detect alterations in microsatellite sequences in colon tumor samples. Ligation profiles on mononucleotide repeat sequences were determined for four related thermostable DNA ligases, Thermus thermophilus ( Tth ) ligase, Thermus sp. AK16D ligase, Aquifex aeolicus ligase and the K294R mutant of the Tth ligase. While the limit of detection for point mutations was one mutation in 1000 wild-type sequences, the ability to detect a single base deletion in a 10 base mononucleotide repeat was one mutation in 100 wild-type sequences. Furthermore, the misligation error increased exponentially as the length of the mono-nucleotide repeat increased, and was 10% of the correct signal for a 19 base mononucleotide repeat. A fluorescent ligase-based assay [polymerase chain reaction/ligase detection reaction (PCR/LDR)] correlated with results obtained using a radioactive assay to detect instability within the TGF-beta Type II receptor gene. PCR/LDR was also used to detect the APCI1307K mononucleotide repeat allele which has a carrier frequency of 6.1% in Ashkenazi Jewish individuals. In a blind study, 30 samples that had been typed for the presence of the APCI1307K allele were tested. The PCR/LDR results correlated with those obtained using sequencing and allele-specific oligonucleotide hybridization for 16 samples carrying the mutation and 13 wild-type samples. Ligation assays that characterize mononucleotide repeats can be used to rapidly detect somatic mutations in tumors, and to screen for individuals who have a hereditary predisposition to develop colon cancer. 相似文献
63.
Ai H Lvov YM Mills DK Jennings M Alexander JS Jones SA 《Cell biochemistry and biophysics》2003,38(2):103-114
A recently developed method for surface modification, layer-by-layer (LbL) assembly, has been applied to silicone, and its
ability to encourage endothelial cell growth and control cell growth patterns has been examined. The surfaces studied consisted
of a precursor, with alternating cationic polyethyleneimine (PEI) and anionic sodium polystyrene sulfonate (PSS) layers followed
by alternating gelatin and poly-d-lysine (PDL) layers. Film growth increased linearly with the number of layers. Each PSS/PEI bilayer was 3 nm thick, and each
gelatin/PDL bilayer was 5 nm thick. All layers were more hydrophilic than the unmodified silicone rubber surface, as determined
from contact angle measurements. The contact angle was primarily dictated by the outermost layer. Of the coatings studied,
gelatin was the most hydrophilic. A film of (PSS/PEI)4/(gelatin/PDL)4/ gelatin was highly favorable for cell adhesion and growth, in contrast to films of (PSS/PEI)8 or (PSS/PEI)8/PSS. Cell growth patterns were successfully controlled by selective deposition of microspheres on silicone rubber, using
microcontact printing with a silicone stamp. Cell adhesion was confined to the region of microsphere deposition. These results
demonstrate that the LbL self-assembly technique provides a general approach to coat and selectively deposit films with nanometer
thickness on silicone rubber. Furthermore, they show that this method is a viable technique for controlling cellular adhesion
and growth. 相似文献
64.
Litvin O Tiunova A Connell-Alberts Y Panchin Y Baranova A 《Journal of cellular and molecular medicine》2006,10(3):613-634
Connexins had been considered to be the only class of the vertebrate proteins capable of gap junction formation; however, new candidates for this function with no homology to connexins, termed pannexins were discovered. So far three pannexins were described in rodent and human genomes: Panx1, Panx2 and Panx3. Expressions of pannexins can be detected in numerous brain structures, and now found both in neuronal and glial cells. Hypothetical roles of pannexins in the nervous system include participating in sensory processing, hippocampal plasticity, synchronization between hippocampus and cortex, and propagation of the calcium waves supported by glial cells, which help maintain and modulate neuronal metabolism. Pannexin also may participate in pathological reactions of the neural cells, including their damage after ischemia and subsequent cell death. Recent study revealed non-gap junction function of Panx1 hemichannels in erythrocytes, where they serve as the conduits for the ATP release in response to the osmotic stress. High-throughput studies produced some evidences of the pannexin involvement in the process of tumorigenesis. According to brain cancer gene expression database REMBRANDT, PANX2 expression levels can predict post diagnosis survival for patients with glial tumors. Further investigations are needed to verify or reject hypotheses listed. 相似文献
65.
Broad host range vectors derived from an RSF1010::Tn1 plasmid 总被引:2,自引:0,他引:2
66.
Taichiro Miyake Kosuke Soda Yasushi Itoh Yoshihiro Sakoda Hirohito Ishigaki Tomoya Nagata Hideaki Ishida Misako Nakayama Hiroichi Ozaki Hideaki Tsuchiya Ryuzo Torii Hiroshi Kida & Kazumasa Ogasawara 《Journal of medical primatology》2010,39(1):58-70
Background Highly pathogenic avian influenza virus (HPAIV) infection has a high mortality rate in humans. Secondary bacterial pneumonia with HPAIV infection has not been reported in human patients, whereas seasonal influenza viruses sometimes enhance bacterial pneumonia, resulting in substantial morbidity and mortality. Therefore, if HPAIV infection were accompanied by bacterial infection, an increase in mortality would be expected. We examined whether a vaccine against HPAIV prevents severe morbidity caused by mixed infection with HPAIV and bacteria.
Methods H7N7 subtype of HPAIV and Streptococcus pneumoniae were inoculated into cynomolgus macaques with or without vaccination of inactivated whole virus particles .
Results Vaccination against H7N7 HPAIV decreased morbidity caused by HPAIV and pneumonia caused by S. pneumoniae . Bacterial replication in lungs was decreased by vaccination against HPAIV, although the reduction in bacterial colonies was not significant.
Conclusions Vaccination against HPAIV reduces pneumonia caused by bacterial superinfection and may improve prognosis of HPAIV-infected patients. 相似文献
Methods H7N7 subtype of HPAIV and Streptococcus pneumoniae were inoculated into cynomolgus macaques with or without vaccination of inactivated whole virus particles .
Results Vaccination against H7N7 HPAIV decreased morbidity caused by HPAIV and pneumonia caused by S. pneumoniae . Bacterial replication in lungs was decreased by vaccination against HPAIV, although the reduction in bacterial colonies was not significant.
Conclusions Vaccination against HPAIV reduces pneumonia caused by bacterial superinfection and may improve prognosis of HPAIV-infected patients. 相似文献
67.
S Nakayama Y Nagashima M Hoshino A Moriyama K Takahashi T Watanabe M Yoshida 《Journal of biochemistry》1983,93(5):1297-1304
To investigate the active site structures of porcine pepsin and Rhizopus chinensis acid protease (RAP), spin label techniques were applied for these enzymes. Comparison of spin labeled porcine pepsin and RAP suggested that the active site cleft of porcine pepsin was narrower at the top, but wider at the bottom than that of RAP. Addition of pepstatin restricted the motion of the labeled nitroxide radicals. Under alkaline conditions, the enzymes changed their conformation discontinuously and irreversibly to open the active site clefts and to lose the binding ability for pepstatin. The denaturation points of both the enzymes were determined to be pH 6.2. 相似文献
68.
Sébastien Lavoué Kouji Nakayama Dean R. Jerry Yusuke Yamanoue Naoki Yagishita Nobuaki Suzuki Mutsumi Nishida Masaki Miya 《Gene》2014
Delineation of the fish family Percichthyidae (Percomorphaceae) has a long and convoluted history, with recent morphological-based studies restricting species members to South American and Australian freshwater and catadromous temperate perches. Four recent nuclear gene-based phylogenetic studies, however, found that the Percichthyidae was not monophyletic and was nested within a newly discovered inter-familial clade of Percomorphaceae, the Centrarchiformes, which comprises the Centrarchidae and 12 other families. Here, we reexamined the systematics of the Percichthyidae and Centrarchiformes based on new mitogenomic information. Our mitogenomic results are globally congruent with the recent nuclear gene-based studies although the overall amount of phylogenetic signal of the mitogenome is lower. They do not support the monophyly of the Percichthyidae, because the catadromous genus Percalates is not exclusively related to the freshwater percichthyids. The Percichthyidae (minus Percalates) and Percalates belong to a larger clade, equivalent to the Centrarchiformes, but their respective sister groups are unresolved. Because all recent analyses recover a monophyletic Centrarchiformes but with substantially different intra-relationships, we performed a simultaneous analysis for a character set combining the mitogenome and 19 nuclear genes previously published, for 22 centrarchiform taxa. This analysis furthermore indicates that the Centrarchiformes are divided into three lineages and the superfamily Cirrhitoidea is monophyletic as well as the temperate and freshwater centrarchiform perch-like fishes. It also clarifies some of the relationships within the freshwater Percichthyidae. 相似文献
69.
A series of fatty acid ethyl esters ranging from C14 to C24 was isolated from a hexane extract of the liverwort Conocephalum conicum, these esters accounted for 77% of the extract. The ethyl esters consisting of even-numbered fatty acids were predominant and ethyl palmitate was the major constituent. 相似文献
70.
Yasuko Nakayama 《Microbiology and immunology》1967,11(2):95-101
The zymogram patterns of esterases and catalases of mycobacterial strains were studied using the thin layer agar electrophoresis. Though there were some variations, Mycobacterium hominis, M. bovis, M. kansasii, M. fortuitum, M. runyonii, M. avium, M. phlei and M. smegmatis seemed to show species-specific patterns consisting of 2 to 6 esterase bands and one or more catalase bands. The patterns of scotochromogens and nonchromogens were rather variable. 相似文献