5 research outputs found
An Iodine-Free and Directed-Disulfide-Bond-Forming Route to Insulin Analogues
An
iodine-free synthetic route to insulin analogues has been established
via a directed disulfide bond formation strategy. This method is completely
compatible with oxidation-sensitive residues. The key step is constructing
the third disulfide bond via a novel procedure involving phenylÂacetylÂaminoÂmethyl
group (Phacm), immobilized Penicillin G Acylase, and Ellman’s
reagent. We expect that this method could be broadly utilized for
synthesizing insulin-like and other cysteine-rich peptides, in particular,
where oxidation-sensitive residues are present in the sequence
Method for Characterization of PEGylated Bioproducts in Biological Matrixes
PEGylation
of peptides and proteins has been widely used to enhance
stability and reduce immunogenicity of biotherapeutics. Characterizing
the degradation of these PEGylated products in biological fluids can
yield essential information to support pharmacokinetic evaluations
and provide clues about their <i>in vivo</i> properties
useful for further molecular optimization. In this paper, we describe
a novel and uncomplicated approach to characterize PEGylated peptides
or proteins and their related degradation products in biological matrixes.
The method involves direct liquid chromatography/mass spectrometry
(LC/MS) analysis of animal sera containing low nanograms to low micrograms
per milliliter of PEGylated product with or without an acetonitrile
precipitation sample treatment. Applying the methodology to analyze
the model PEGylated peptides, 20K PEGylated-Pancreatic Polypeptide
analogue (PPA) and 20K PEGylated-glucagon, we elucidated the decomposition
pathways occurring in animal sera. The data provided direct evidence
of cleavages within the peptide backbone. The identified degradation
products were unambiguously confirmed by tandem mass spectrometry
with high-energy C-trap dissociation (HCD) analysis, followed with
in-source fragmentation. Additional spiking studies demonstrated nearly
full recovery of PEGylated products, linear detection when the spiked
concentration of PEGylated product was ≤1000 ng/mL, and a low
ng/mL limit of quantitation (LOQ)
Additional file 4: of Telomerase regulation by the long non-coding RNA H19 in human acute promyelocytic leukemia cells
Figure S2. Validation of the anti-hTERT antibody specificity. Immunoprecipitation of the hTERT/hTR complex was performed using an anti-hTERT antibody (Rockland) or pre-immune IgG as described in Material and Methods. The presence of hTR was detected by quantitative RT-PCR. Results were expressed as means +/− SEM. t-test *p < 0.05. (PDF 90 kb
Additional file 2: of Telomerase regulation by the long non-coding RNA H19 in human acute promyelocytic leukemia cells
Table S2. Data analysis of the microarray experiments. Lists of genes with significant differential expression levels (fold changes of ±2 fold; p < 0.01). Some details correspond to the results displayed in Venn diagrams presented in Fig. 1c of the main text. (XLSX 66 kb
Table_1_Methylprednisolone is related to lower incidence of postoperative bleeding after flow diverter treatment for unruptured intracranial aneurysm.DOCX
Background and objectivesRegarding the anti-inflammatory effect, methylprednisolone is a candidate to prevent patients with unruptured intracranial aneurysms (UIAs) from postoperative bleeding (PB) after flow diverter (FD) treatment. This study aimed to investigate whether methylprednisolone is related to a lower incidence of PB after FD treatment for UIAs.MethodsThis study retrospectively reviewed UIA patients receiving FD treatment between October 2015 and July 2021. All patients were observed until 72  h after FD treatment. The patients receiving methylprednisolone (80  mg, bid, for at least 24 h) were considered as standard methylprednisolone treatment (SMT) users, otherwise as non-SMT users. The primary endpoint indicated the occurrence of PB, including subarachnoid hemorrhage, intracerebral hemorrhage, and ventricular bleeding, within 72 h after FD treatment. This study compared the incidence of PB between SMT users and non-SMT users and investigated the protective effect of SMT on PB after FD treatment using the Cox regression model. Finally, after controlling the potential factors related to PB, we performed subgroup analysis to further confirm the protective effect of SMT on PB.ResultsThis study finally included 262 UIA patients receiving FD treatment. PB occurred in 11 patients (4.2%), and 116 patients (44.3%) received SMT postoperatively. The median time from the end of surgery to PB was 12.3 h (range: 0.5–48.0 h). SMT users had a lower incidence of PB comparing with non-SMT users (1/116, 0.9% vs. 10/146, 6.8%, respectively; p = 0.017). The multivariate Cox analysis demonstrated that SMT users (HR, 0.12 [95%CI, 0.02–0.94], p = 0.044) had a lower risk of PB postoperatively. After controlling the potential factors related to PB (i.e., gender, irregular shape, surgical methods [FD and FD + coil] and UIA sizes), the patients receiving SMT still had a lower cumulative incidence of PB, comparing with patients receiving non-SMT (all p ConclusionSMT was correlated with the lower incidence of PB for patients receiving FD treatment and may be a potential method to prevent PB after the FD treatment.</p