inflammation the danger signal and autoimmunity

... proteins and disease Therapeutic implications of sHSPs and other molecular chaperones Temperature induced synthesis of sHSPs protects against ischemia ⁄ reperfusion damage to the heart, brain, and ... and drugs The therapeutic application of sHSPs is further suggested by study of other molecular chaperones, with disruption of HSPs that protect deregulated intracellular signaling proteins and ... post-translational changes, and these are outlined below and described in the text of the review In addition, changes in the amounts of sHSPs, unaccompanied by a structural change in the protein per se,...

Ngày tải lên: 19/02/2014, 18:20

... temperature with the addition of 400 lL 5% polyethylenimine and 800 lL 200 mM dithiothreitol for 10 The sample was then centrifuged at 35 000 g for h at °C The supernatant was decanted and the insoluble ... beginning at the time of heat shock, plated and CFU counted The proportions of viable cells expressing the pET 16b-MTB HSP 16.3 vector and the two control vectors were plotted for 0, and h following ... was discarded The supernatant was then ready for purification using the Pharmacia FPLC system The supernatant containing the soluble protein was filtered through a 0.22 lm filter and was loaded...

Ngày tải lên: 23/03/2014, 21:21

... mM The samples were mixed with the sample buffer and after boiling were subjected to SDS gel electrophoresis [22] After staining, the gels were scanned and evaluated by the ONEDSCAN program The ... trypsinolysis under the same conditions, the band of actin disappeared in the first 15–30 s (Fig 4B, curve 2) and a number of faint bands with different molecular masses were accumulated in the incubation ... cooling and diluting with buffer G, so that the concentration of actin becomes equal to 10 lM, the reaction was started by the addition of KCl and MgCl2 up to the final concentrations 50 and mM,...

Ngày tải lên: 20/02/2014, 23:20

... PAGE and SEC indicated the class I and II sHsps have different oligomeric structures, neither of these techniques are primary methods for size determination Therefore, to better understand the ... for 15 at 16 250 g and the supernatant fractions removed The supernatant and pellet fractions were treated with SDS sample buffer and the entire amount analyzed by SDS/PAGE and Coomassie blue ... nm Therefore, the relative sizes of the two oligomers are consistent with their behavior on nondenaturing PAGE and SEC Their appearance is also similar to what has been observed for class I and...

Ngày tải lên: 07/03/2014, 15:20

... indicated by the dotted lines assemblies A transfer of CS and HspH into the void volume, at the expense of the other fractions, was confirmed by SDS-PAGE (Fig 7C) Neither HspH nor CS shifted into the void ... the secondary structure The traces recorded are typical of b-strand proteins and show a single minimum elipticity in the range of 217 nm, similar to that observed for other sHsps [14,18,32] The ... the major species in the spectrum Representative charge states of the associated oligomers (XmerY+) are shown NanoESI-MS was used to confirm the effects of concentration and temperature upon the...

Ngày tải lên: 23/03/2014, 12:20

... with the RIKEN mAb anti-(aB-crystallin) (a) or the monoclonal antibody to SC35 (d) and costained with anti-Sm (b and e) The yellow pseudocolour shows the extent of colocalization between the two ... nuclear bodies, but the combination S19D/ S45D had the strongest effect The largest number of positive cells was obtained with the S19D/S45D/S59D mutant These results confirm the correlation between ... that the speckle staining is indeed due to the presence of phosphorylated aB-crystallin, we performed Western blotting with the anti(aB-crystallin) and anti-S45p IgGs on the isolated nuclei of these...

Ngày tải lên: 23/03/2014, 13:20

... examined the role of residues in the C-terminal extensions of other sHsps, notably aA- and aB-crystallin, the flexible regions of these proteins are unique and distinct from that of Hsp25 The a-crystallins ... in the environment of the tryptophan residues resulting from the mutations The tryptophan residues of Hsp25 are located in the N-terminal domain at positions 16, 22, 43, 46 and 52, and in the ... precipitate sinking to the bottom of the cuvette and therefore not obscuring the light path [42] Thus, the Q194A mutant showed thermostability similar to that of wildtype Hsp25, whereas the glutamic acid...

Ngày tải lên: 07/03/2014, 04:20

... between strands b2 and b6 of neighboring monomers The p26 modifications examined in this study are not located in either of these strands and they are not considered further As the basic p26 oligomer ... monomers and their b-strand elements in the a-crystallin domain, the amino acid substitution F112R introduces two changes at the dimer–dimer interface Although Y116 and R114 are in b-strand 7, neither ... in the presence of dithiothreitol, and solution turbidity was measured at 400 nm The p26 variants tested are indicated in the figure and they appear in the same order in each histogram group The...

Ngày tải lên: 07/03/2014, 12:20

... sHsp The primary structure of the a-crystallin domain of human sHsp is very conservative and the loop connecting the b5 and b7 strands is shorter than the corresponding loop connecting the b5 and ... However, the overall structures of aB-crystallin and HSP27 predicted by these two models are very similar, and the positions of the b-strands correlate well with the corresponding positions of the ... databank accession code 1GME) The N- and C-terminal domains are indicated by N and C correspondingly All b-strands are numbered and the b5 and b7 strands are shown in red and blue, respectively G104...

Ngày tải lên: 23/03/2014, 07:20

... co-exist in many cells and under different circumstances and that they are capable of interacting with each other The physical and functional interactions may therefore provide another layer of control ... knockout of Hsf1 and Hsf4, the abnormal levels of FGF-7 returned to normal, and proliferation and differentiation of the epithelial cells were stabilized These findings indicate that HSF1 and HSF4 compete ... in the olfactory epithelium warrants further investigations In accordance with the fundamental role of HSF1 in the heat shock response, the requirement of HSF1 and HSF4 in development of the...

Ngày tải lên: 18/02/2014, 04:20

... Hence, these observations support the hypothesis that the expression of expanded httEx1 and the presence of httEx1 aggregates ⁄ inclusion bodies correlate with elevated ROS levels, and that NAC and ... analyzed The arrow indicates the position of the more intensively oxidized polypeptide in the assay The bracket underlines the domain (molecular mass range 10–40 kDa) of the oxyblots that contains the ... problem that the cell has to challenge to promote the formation and clearance of these bodies Aggregate clearance could therefore be balanced between the rate of huntingtin aggregation and the deleterious...

Ngày tải lên: 19/02/2014, 06:20

... constructs and pET21(+) were digested with BamHI and XhoI before electrophoresis in 1% agarose gels Linearized pET21(+) and p26 cDNAs were excised and purified with the GFXTM PCR DNA and Gel Band Purification ... transferred to nitrocellulose, and reacted with antibody to p26 using the ECL procedure The top of each gradient is to the left of the figure, and numbers across the top indicate successive samples ... gels, transferred to nitrocellulose, and reacted with antibody to the His6 epitope tag The top of each gradient is to the left of the figure, and numbers across the top indicate successive samples...

Ngày tải lên: 22/02/2014, 04:20

... oligomers of Hsp20 and its S16D mutant to monomers that tend to unfold and aggregate These aggregates are retarded on the top of the column and therefore not detected on the chromatogram The data presented ... of mgÆmL)1, and then stored on ice for h After incubation, the samples were diluted sixfold in the same buffer, minus urea and b-mercaptoethanol, and dialyzed against two changes of the same buffer ... L of the E coli culture When the S16D mutant of Hsp20 was expressed in E coli, most of the protein was insoluble in the lysis buffer and this buffer extracted less than 20% of the protein The...

Ngày tải lên: 07/03/2014, 15:20

... during the course of F-actin thermal denaturation All the methods used here show that the size of these complexes depends on the Hsp27-3D ⁄ actin ratio in the initial mixture of Hsp27-3D and F-actin ... 68 °C in the absence or in the presence of Hsp27-3D (0.25–1.0 mgÆmL)1) Under these conditions, F-actin was fully denatured both in the absence and in the presence of Hsp27-3D [2] In the absence ... 0.015 to 0.5 mgÆmL)1, and the F-actin concentration was constant and equal to 0.5 mgÆmL)1 Other conditions were the same as in Fig 1A The Rh values were determined from the Rh distributions obtained...

Ngày tải lên: 16/03/2014, 06:20

... extension and another in the C-terminus The deletion of these motifs, termed G (multiple glycine), R (multiple arginine), and TS (multiple threonine ⁄ serine), was confirmed by sequencing and the modified ... emission intensities, with the peak of G and TS at 348 nm and R at 360 nm The fluorescence intensity of R was the lowest and it was red-shifted in comparison to other samples The extent of tertiary ... enriched in glycine and the other in arginine Additionally, the C-terminus contains an unusual stretch of residues endowed with threonine and serine These regions are interesting because the characteristics...

Ngày tải lên: 30/03/2014, 20:20

... activity and oligomerization [44] In a-crystallin, on the other hand, numerous point mutations and even truncation of the first half of the N-terminal region did not affect complex formation and functionality ... isoleucine residues Two further constructs, HspH(D5C) and HspH(D15C), contained the entire I-X-I motif To assess the role of the conserved motif in the context of the full-length protein, three ... one of the three main consensus regions of the a-Hsp family [5] There is little experimental evidence to circumstantiate the role of the C-terminal extension, and even less concerning the conserved...

Ngày tải lên: 31/03/2014, 21:21

... of the immune system to promote tumorigenesis The ancient Roman physician Galen (129 – 199 C.E.) was the first to posit the causal relationship between cancer and inflammation In 1863, the "Father ... protein signal cascade Heat-shock protein signal cascade Necrotic tumorderived mammalian gp96 and HSP70 can signal antigen-presenting cells (APCs) via CD14, and other receptors such as TLRs and CD91 ... neoplasia and cervical cancer [92] These examples begin to unveil the complex relationship between HSPs and cancer formation Microbes, inflammation, heat-shock proteins and cancer The parasitic...

Ngày tải lên: 10/08/2014, 22:20

... superficial and interstitial hyperthermia and figures 2-8 (a), and (b) show the superficial microwave hyperthermia, and interstitial hyperthermia, respectively In regional hyperthermia, the heat ... tissue, and photocoagulation in the blood vessels due to the high power of the laser and the difficulty in controlling the local temperature (Fig 2-6) [38] Therefore, a new modality for the biologically ... Investigation of the physical contribution of Néel and Brown relaxation loss power to the AC heating power of fluidic SPNPs e Understand the physical mechanisms of the AC heating and the crucial physical...

Ngày tải lên: 09/09/2015, 10:08

... assistance and comforting words To all the members and colleagues in the lab, for accompanying me through out the course of my study To Dr Leslie, Jeremy, James and Sean from the AHU, and Ms Huang ... for their help in the animal studies To Karen, Weiping and Huang Bo, for their assistance in DNA sequencing To Dr Lu and Jason, for their help and discussion in TLR4 experiments To Soon Yew and ... their wonderful help, advice and friendship To Paul, Sherry, Jia Ling, Yong Mei, Annette, Qian Feng and Ying Ying, for their encouragement and help in one way or the other To my aunt, uncle and...

Ngày tải lên: 09/10/2015, 11:06