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101.
Amrita A. Nagle Fei-Fei Gan Gavin Jones Choon-Leng So Geoffrey Wells Eng-Hui Chew 《PloS one》2012,7(11)
Multifunctional trans-cinnamaldehyde (CA) and its analogs display anti-cancer properties, with 2-benzoyloxycinnamaldehyde (BCA) and 5-fluoro-2-hydroxycinnamaldehyde (FHCA) being identified as the ortho-substituted analogs that possess potent anti-tumor activities. In this study, BCA, FHCA and a novel analog 5-fluoro-2-benzoyloxycinnamaldehyde (FBCA), were demonstrated to decrease growth and colony formation of human colon-derived HCT 116 and mammary-derived MCF-7 carcinoma cells under non-adhesive conditions. The 2-benzoyloxy and 5-fluoro substituents rendered FBCA more potent than BCA and equipotent to FHCA. The cellular events by which these cinnamaldehydes caused G2/M phase arrest and halted proliferation of HCT 116 cells were thereby investigated. Lack of significant accumulation of mitosis marker phospho-histone H3 in cinnamaldehyde-treated cells indicated that the analogs arrested cells in G2 phase. G2 arrest was brought about partly by cinnamaldehyde-mediated depletion of cell cycle proteins involved in regulating G2 to M transition and spindle assembly, namely cdk1, cdc25C, mad2, cdc20 and survivin. Cyclin B1 levels were found to be increased, which in the absence of active cdk1, would fail to drive cells into M phase. Concentrations of cinnamaldehydes that brought about dysregulation of levels of cell cycle proteins also caused tubulin aggregation, as evident from immunodetection of dose-dependent tubulin accumulation in the insoluble cell lysate fractions. In a cell-free system, reduced biotin-conjugated iodoacetamide (BIAM) labeling of tubulin protein pretreated with cinnamaldehydes was indicative of drug interaction with the sulfhydryl groups in tubulin. In conclusion, cinnamaldehydes treatment at proapoptotic concentrations caused tubulin aggregation and dysegulation of cell cycle regulatory proteins cdk1 and cdc25C that contributed at least in part to arresting cells at G2 phase, resulting in apoptotic cell death characterized by emergence of cleaved forms of caspase 3 and poly (ADP-ribose) polymerase (PARP). Results presented in this study have thus provided further insights into the intricate network of cellular events by which cinnamaldehydes induce tumor cell death. 相似文献
102.
Amrita Banerjee Arijit Jana Bikash R. Pati Keshab C. Mondal Pradeep K. Das Mohapatra 《The protein journal》2012,31(4):306-327
The tannase protein sequences of 149 bacteria and 36 fungi were retrieved from NCBI database. Among them only 77 bacterial
and 31 fungal tannase sequences were taken which have different amino acid compositions. These sequences were analysed for
different physical and chemical properties, superfamily search, multiple sequence alignment, phylogenetic tree construction
and motif finding to find out the functional motif and the evolutionary relationship among them. The superfamily search for
these tannase exposed the occurrence of proline iminopeptidase-like, biotin biosynthesis protein BioH, O-acetyltransferase,
carboxylesterase/thioesterase 1, carbon–carbon bond hydrolase, haloperoxidase, prolyl oligopeptidase, C-terminal domain and
mycobacterial antigens families and alpha/beta hydrolase superfamily. Some bacterial and fungal sequence showed similarity
with different families individually. The multiple sequence alignment of these tannase protein sequences showed conserved
regions at different stretches with maximum homology from amino acid residues 389–469 and 482–523 which could be used for
designing degenerate primers or probes specific for tannase producing bacterial and fungal species. Phylogenetic tree showed
two different clusters; one has only bacteria and another have both fungi and bacteria showing some relationship between these
different genera. Although in second cluster near about all fungal species were found together in a corner which indicates
the sequence level similarity among fungal genera. The distributions of fourteen motifs analysis revealed Motif 1 with a signature
amino acid sequence of 29 amino acids, i.e. GCSTGGREALKQAQRWPHDYDGIIANNPA, was uniformly observed in 83.3 % of studied tannase
sequences representing its participation with the structure and enzymatic function. 相似文献
103.
Laura J. Marinelli Mariana Piuri Zuzana Swigoňová Amrita Balachandran Lauren M. Oldfield Julia C. van Kessel Graham F. Hatfull 《PloS one》2008,3(12)
Advances in DNA sequencing technology have facilitated the determination of hundreds of complete genome sequences both for bacteria and their bacteriophages. Some of these bacteria have well-developed and facile genetic systems for constructing mutants to determine gene function, and recombineering is a particularly effective tool. However, generally applicable methods for constructing defined mutants of bacteriophages are poorly developed, in part because of the inability to use selectable markers such as drug resistance genes during viral lytic growth. Here we describe a method for simple and effective directed mutagenesis of bacteriophage genomes using Bacteriophage Recombineering of Electroporated DNA (BRED), in which a highly efficient recombineering system is utilized directly on electroporated phage DNA; no selection is required and mutants can be readily detected by PCR. We describe the use of BRED to construct unmarked gene deletions, in-frame internal deletions, base substitutions, precise gene replacements, and the addition of gene tags. 相似文献
104.
Xu Y Yasin A Tang R Scharer JM Moo-Young M Chou CP 《Applied microbiology and biotechnology》2008,81(1):79-87
Functional expression of lipase B from Pseudozyma antarctica (PalB) in the cytoplasm of Escherichia coli BL21(DE3) and its mutant derivative Origami B(DE3) was explored. Coexpression of DsbA was found to be effective in enhancing
PalB expression. The improvement was particularly pronounced with Origami B(DE3) as a host, suggesting that both folding and
disulfide bond formation may be major factors limiting PalB expression. Fusion tag technique was also explored by constructing
several PalB fusions for the evaluation of their expression performance. While the solubility was enhanced for most PalB fusions,
only the DsbA tag was effective in boosting PalB activity, possibly by both enhanced solubility and correct disulfide bond
formation. Our results suggest that PalB activity is closely associated with correct disulfide bond formation, and increased
solubilization by PalB fusions does not necessarily result in activity enhancement. 相似文献
105.
Yali Xu Amrita Yasin Thomas Wucherpfennig C. Perry Chou 《World journal of microbiology & biotechnology》2008,24(12):2827-2835
Functional expression of heterologous Pseudozyma antarctica lipase B (PalB) in the periplasm of Escherichia coli was explored using four fusion tags, i.e. DsbC, DsbA, maltose-binding protein (MBP), and FLAG in the sequence of increasing
expression efficacy. Amongst these fusion tags, FLAG and MBP appear to be the most effective ones in terms of boosting enzyme
activity and enhancing solubility of PalB, respectively. Overexpression of these PalB fusions often resulted in concomitant
formation of insoluble inclusion bodies. Coexpression of a selection of periplasmic folding factors, including DegP (and its
mutant variant of DegPS210A), FkpA, DsbA, DsbC, and a cocktail of SurA, FkpA, DsbA, and DsbC, could improve the expression performance. Coexpression
of DsbA appeared to be the most effective in reducing the formation of inclusion bodies for all the four PalB fusions, implying
that functional expression of PalB could be limited by initial bridging of disulfide bonds. Culture performance was optimized
by overexpressing FLAG-PalB with DsbA coexpression, resulting in a high volumetric PalB activity of 360 U/L. 相似文献
106.
Cai ZW Wei D Borzilleri RM Qian L Kamath A Mortillo S Wautlet B Henley BJ Jeyaseelan R Tokarski J Hunt JT Bhide RS Fargnoli J Lombardo LJ 《Bioorganic & medicinal chemistry letters》2008,18(4):1354-1358
Introduction of the 2,4-difluoro-5-(cyclopropylcarbamoyl)phenylamino group at the C-4 position of the pyrrolo[2,1-f][1,2,4] triazine scaffold led to the discovery of a novel sub-series of inhibitors of VEGFR-2 kinase activity. Subsequent SAR studies on the 1,3,5-oxadiazole ring appended to the C-6 position of this new sub-family of pyrrolotriazines resulted in the identification of low nanomolar inhibitors of VEGFR-2. Antitumor efficacy was observed with compound 37 against L2987 human lung carcinoma xenografts in athymic mice. 相似文献
107.
The present study was aimed at assessing the effects of the unusual schedules of shift-working men on anxiety and mental health of their spouses and children. The anxiety and mental health of 396 subjects that include 133 spouses and 263 children of day-working and shift-working men were monitored by employing specific inventory. Anxiety scales (trait, state and free-floating) and mental health inventory were administrated to each subject. Results indicate that the average levels of free-floating anxiety for the spouses and children of shift-working men were found to be significantly higher in comparison with that of their counterparts sampled in the family of day workers. This was also true when data were segregated with respect of gender and compared. Except in female children of shift workers all other groups of children had statistically significantly higher level of state anxiety as compared with that of their respective mothers. The results of this study further reveal that the spouses of shift-working men exhibited lower mental health rating as compared with the spouses of day-working men. Furthermore, a significant negative relationship was noticed between anxiety and mental health in all cases excepting in spouses of shift-working men for state anxiety. Although the findings of the present study could be attributed to the irregular routine of shift workers, it appears mandatory to carryout further intensive studies to draw convincing conclusions. 相似文献
108.
We studied case construction behavior of bagworm moth, Eumeta crameri (Lepidoptera: Psychidac) in the host plant, Acacia nilotica, at two sites. At the time of the study, Site A had 1-year-old A. nilotica saplings only, whereas Site B had full grown trees. The larva of bagworm moth used either thorns or cut-twigs or both as the materials for building its case. It renovated its case three times, and during each instance it increased the volume of its case by replacing older thorns or cut-twigs by newer and longer ones. However, it exhibited a spectacular predilection for thorns, irrespective of the sites, during the first-instar stage of its development. Thereafter, at Site A it used exclusively cut-twigs, whereas at Site B it preferably used thorns provided they were available in the range of the required length and within the threshold distance. It appears that the bagworm has an ability to process thorn length and distance signals hierarchically to fulfill its priority, that is, case building. Thus it optimizes time and energy expenditures during the period of its larval growth leading to pupation by toggling its preference between thorns and cut-twigs. 相似文献
109.
110.