32 research outputs found
Evaluation of genipin-crosslinked chitosan hydrogels as a potential carrier for silver sulfadiazine nanocrystals
In the present study genipin crosslinked chitosan (CHI) hydrogels, which had been
constructed and reported in our previous studies (Lei Gao, et al. Colloids Surf. B
Biointerfaces. 2014, 117: 398), were further evaluated for their advantage as a carrier
for silver sulfadiazine (AgSD) nanocrystal systems. Firstly, AgSD nanocrystals with a
mean particle size of 289 nm were prepared by wet milling method and encapsulated
into genipin crosslinked CHI hydrogels. AgSD nanocrystals displayed a uniform
distribution and very good physical stability in the hydrogel network.
Swelling-dependent release pattern was found for AgSD nanocrystals from hydrogels
and the release profile could be well fitted with Peppas equation. When AgSD
nanocrystals were encapsulated in hydrogels their fibroblast cytotoxicity decreased
markedly, and their antibacterial effects against Staphylococcus aureus, Escherichia
coli and Pseudomonas aeruginosa were still comparable to unencapsulated AgSD
nanocrystals. In vivo evaluation in excision and burn cutaneous wound models in
mice showed that AgSD nanocrystal hydrogels markedly decreased the expression of
inflammatory cytokine IL-6, but increased the levels of growth factors VEGF-A and
TGF-β1. Histopathologically, the wounds treated by hydrogels containing AgSD
nanocrystals showed the best healing state compared with commercial AgSD cream,
hydrogels containing AgSD bulk powders and blank hydrogels. The wounds treated
by AgSD nanocrystal hydrogels were dominated by marked fibroblast proliferation,
new blood vessels and thick regenerated epithelial layer. Sirius Red staining assay
indicated that AgSD nanocrystal hydrogels resulted in more collagen deposition
characterized by a large proportion of type I fibers. Our study suggested that
genipin-crosslinked CHI hydrogel was a potential carrier for local antibacterial
nanomedicines
Peptide Hydrolysis by Metal-Cyclen Complexes and Their Analogues: Insights from Theoretical Studies
In
the present DFT study, mechanisms of peptide hydrolysis by CoÂ(III)-
and CuÂ(II)-containing complexes of 1,4,7,10-tetraazacyclododecane
(cyclen), <b>1-Co</b> and <b>1-Cu</b>, respectively, and
1-oxa-4,7,10-triazacyclododecane (oxacyclen), <b>2-Co</b> and <b>2-Cu</b>, respectively, and their analogues have been investigated.
In addition, the effects of the ligand environment, pendant (an organic
group containing a recognition site) and metal ion (CoÂ(III), CuÂ(II),
NiÂ(II), ZnÂ(II), CdÂ(II), and PdÂ(II)), on the energetics of this reaction
have been elucidated. The reactant of the <b>1-Co</b> complex
exists in the <i>syn–anti</i> conformation, while
that of <b>1-Cu</b> in the <i>syn–syn</i> form.
For both these complexes, stepwise and concerted mechanisms were found
to occur with similar barriers. The substitution of one of the nitrogen
atoms in the cyclen macrocycle to create oxacyclen should occur at
position 10 in the CoÂ(III) case and at position 4 in the CuÂ(II) case.
A comparison between the barriers using the common conformation (<i>syn–anti</i>) of <b>1-Co</b> and <b>2-Co</b> showed that both complexes hydrolyze the peptide bond with similar
barriers, i.e., 39.8 kcal/mol for the former and 40.1 kcal/mol for
the latter. This result is in line with the measured data that suggest
that the oxacyclen complex exhibits just four times greater activity
than the cyclen complex. The removal of the pendant (−C<sub>2</sub>H<sub>5</sub>) group in the CoÂ(III)- and CuÂ(II)-cyclen complexes
(<b>1</b>′<b>-Co</b> and <b>1</b>′<b>-Cu</b>, respectively) reduced the barriers by 9.3 and 3.0 kcal/mol,
respectively. For <b>1</b>′<b>-Co</b>, the barrier
of 30.5 kcal/mol is in agreement with the experimental value of 25.9
kcal/mol for the cleavage of myoglobin at pH 9.0 and 50 °C. The
reactants of <b>1</b>′<b>-Cu</b>,<b> 1</b>′<b>-Zn</b>,<b> 1</b>′<b>-Pd</b>,
and <b>1</b>′<b>-Cd</b> adopt the <i>syn–syn</i> conformation, whereas <b>1</b>′<b>-Ni</b> and <b>1</b>′<b>-Co</b> exist in the <i>syn–anti</i> geometry. The barriers for <b>1</b>′<b>-Ni</b> (triplet spin state), <b>1</b>′<b>-Cu</b> (doublet
spin state), <b>1</b>′<b>-Cd</b> (singlet spin
state), <b>1</b>′<b>-Co</b> (singlet spin state),
and <b>1</b>′<b>-Zn</b> (singlet spin state) are
similar, i.e., 27.2, 29.7, 30.5, 30.5, and 31.9 kcal/mol, respectively,
and the highest barrier (41.5 kcal/mol) is computed for <b>1</b>′<b>-Pd</b> (singlet spin state)
Mechanistic Insights into Metal (Pd<sup>2+</sup>, Co<sup>2+</sup>, and Zn<sup>2+</sup>)−β-Cyclodextrin Catalyzed Peptide Hydrolysis: A QM/MM Approach
In this study, mechanistic insights
into the hydrolysis of an extremely
stable tertiary peptide bond (Ser–Pro) in the Ser-Pro-Phe sequence
by an artificial enzyme, metal (Pd<sup>2+</sup>, Co<sup>2+</sup>,
or Zn<sup>2+</sup>)−β-cyclodextrin (CD) complex, have
been provided. In particular, the exact reaction mechanism, the location
of CD (number of −CH<sub>2</sub> groups downstream from the
metal center), conformation of CD (primary or secondary rim of CD
facing the substrate), the number of CD (one or two), and the optimum
metal ion (Pd<sup>2+</sup>, Co<sup>2+</sup>, or Zn<sup>2+</sup>) have
been suggested using a state-of-the-art hybrid quantum mechanics/molecular
mechanics (QM/MM: B3LYP/Amber) approach. The QM/MM calculations suggest
that the internal delivery mechanism is the most energetically feasible
for the peptide hydrolysis. The inclusion of a CD ring at two CH<sub>2</sub> groups downstream from the metal center can provide 3 ×
10<sup>5</sup> times acceleration in the activity, while the replacement
of Pd<sup>2+</sup> with Co<sup>2+</sup> enhances the rate activity
another 3.7 × 10<sup>4</sup> times
Correlation between <i>Txnrd1</i> gene expression and audiological measurements.
<p><sup>*</sup> denotes statistical significance at 0.05 level;</p><p><sup>**</sup> at the 0.01 level;</p><p><sup>***</sup> at the 0.001 level.</p
DPOAE amplitudes correlation with gene expression A) and B): The correlations between <i>Gxp6</i> signal log ratio and DPOAE amplitudes at mid and high frequencies, are two examples of the significant correlations of gene expression changes with DPOAE test results.
<p>C) and D): The correlations between <i>Txnrd1</i> signal log ratio and DPOAE amplitudes at low and mid frequencies were significant.</p
ANOVA results of genechip signal log ratios between groups and average fold changes of validated genes.
<p><sup>*</sup> denotes statistical significance at 0.05 level.</p
ABR thresholds correlation with gene expression A) and B): The correlations between <i>Gxp6</i> qPCR fold changes and ABR thresholds at 12 kHz and 48 kHz are two examples of the significant correlations of gene expression changes with ABR test results.
<p>C) and D): The correlations between <i>Txnrd1</i> signal log ratio and ABR thresholds at 12 kHz and 48 kHz were significant.</p
Correlation between Gpx6 gene expression and audiological measurements.
<p><sup>*</sup> denotes statistical significance at 0.05 level;</p><p><sup>**</sup> at the 0.01 level.</p
A): For both GeneChip and real-time PCR, fold changes of <i>Hspb1</i> gene expression in the cochleae of middle age, old mild hearing loss, and old severe hearing loss groups showed upregulation with age and hearing loss.
<p>B): For both GeneChip and real-time PCR, fold changes of <i>Gpx</i>6 gene expression in cochlea samples showed upregulation in all age groups compared to the young group. C): For both GeneChip and real-time PCR, fold changes of <i>Txnrd1</i> gene expression in the cochleae of middle age, old mild hearing loss, and old severe hearing loss groups showed downregulation with age and hearing loss.</p
Additional file 1: Figure S1. of DNA hypomethylation of a transcription factor binding site within the promoter of a gout risk gene NRBP1 upregulates its expression by inhibition of TFAP2A binding
Work flow diagram. Figure S2. Serum uric acid is regulated by B1 methylation level and NRBP1 expression. a A marginal significant negative association between DNA methylation of B1 and serum uric acid (P value = 0.08). b A significant positive association between NRBP1 expression and serum uric acid (P value = 0.03). Figure S3. Decreased DNA methylation at the promoter region of NRBP1 in gout patients. a The DNA sequence at the promoter region of NRBP1 gene. The CpG sites, designated as B1 to B6, are highlighted in red. b The methylation level for each CpG site indicated in a, was investigated by bisulfite pyrosequencing. Data are represented as mean ± SEM. (*P value <0.01, **P value <0.001, Student’s t test, unpaired, two-sided). (PPTX 1077 kb