Biomedical subjects

Hua Wu

Publications and source records attributed to Hua Wu.

64 records · Page 4Linked to original sources

PubMed2002

Identification of phosphorylated residues that affect the activity of the mitotic kinase Aurora-A.

The activity of the kinase Aurora-A (Aur-A) peaks during mitosis and depends on phosphorylation by one or more unknown kinases. Mitotic phosphorylation sites were mapped by mass spec sequencing of recombinant Aur-A protein that had been activated by incubation in extracts of metaphase-arrested Xenopus eggs. Three sites were identified: serine 53 (Ser-53), threonine 295 (Thr-295), and serine 349 (Ser-349), which are equivalent to Ser-51, Thr-288, and Ser-342, respectively, in human Aur-A. To ask how phosphorylation of these residues might affect kinase activity, each was mutated to either alanine or aspartic acid, and the recombinant proteins were then tested for their ability to be activated by M phase extract. Mutation of Thr-295, which resides in the activation loop of the kinase, to either alanine or aspartic acid abolished activity. The S349A mutant had slightly reduced activity, indicating that phosphorylation is not required for activity. The S349D mutation completely blocked activation, suggesting that Ser-349 is important for either the structure or regulation of Aur-A. Finally, like human Aur-A, overexpression of Xenopus Aur-A transformed NIH 3T3 cells and led to tumors in nude mice. These results provide further evidence that Xenopus Aur-A is a functional ortholog of human Aur-A and, along with the recently described crystal structure of human Aur-A, should help in future studies of the mechanisms that regulate Aur-A activity during mitotic progression.

3T3 Cells↗

PubMed2002

Effects of nociceptin (13-17) in pain modulation at supraspinal level in mice.

This work was designed to observe the effects of nociceptin(13-17), one of the main metabolites of nociceptin (also termed orphanin FQ), in pain modulation at supraspinal level in mice. Intracerebroventricular (i.c.v.) administration of nociceptin/orphanin FQ(13-17) (N/OFQ(13-17)) (5, 0.5, 0.05, 0.005 nmol/mouse) dose-dependently induced potent hyperalgesic effects in the 48 degrees C warm-water tail-flick test in mice. I.c.v. pretreatment with N/OFQ(13-17) (5, 0.5, 0.05 nmol/mouse) potentiated the analgesic effects induced by morphine (i.p., 2 mg/kg) and reversed the hyperalgesic effects induced by N/OFQ (i.c.v., 5 nmol/mouse). The hyperalgesic effects induced by N/OFQ(13-17) could not be antagonized by [Nphe((1))]N/OFQ(1-13)NH((2)) or naloxone. These findings suggest that N/OFQ(13-17) may play important roles in pain modulation at supraspinal level in mice and elicits these effects through a novel mechanism independent of the N/OFQ receptor and the mu, delta and kappa opioid receptors.

Analgesia↗

PubMed2002

The relationship between (99m)Tc-MIBI uptakes and tumor cell death/proliferation state under irradiation.

To evaluate the potential of 99mTc-MIBI imaging to monitor cellular viability of tumor post-irradiation, Ehrlich carcinoma tissues were exposed to 60Co at different single dose (0, 5, 10, 15 and 20 Gy, respectively) and the following protocol were performed at 6 h pre-irradiation and 24, 72 and 144 h post-irradiation, respectively: (1) 99mTc-MIBI planar scintigraphy was performed. Uptake of 99mTc-MIBI was expressed as differential uptake ratio (DUR) and tumor-to-non-target ratio (T/NT). (2) Apoptosis index (AI), percent of necrosis area (PNA) and Proliferating cell nuclear antigen integral absorbance (PCNA-IA)were measured. DUR and T/NT decreased with radiation dose escalating and post-irradiation time prolonging. At 24 h post-irradiation, DUR or T/NT was inversely correlated with AI or PNA (r=-0.849, -0.829, -0.883, -0.855, respectively, n=33, P<0.01) and positively correlated with PCNA-IA (r=0.789, 0.742, respectively, n=33, P<0.01). At 72 and 144 h post-irradiation, DUR or T/NT was only inversely correlated with PNA (r=-0.967, -0.956, -0.915, -0.886, respectively, n=33, P<0.01). It is suggested that 99mTc-MIBI uptake of tumor cell correlated with the changes of cell viability after irradiation. 99mTc-MIBI imaging may be a potential method to monitor tumor cell viability or therapeutic response after irradiation.

Animals↗

PubMed2002

Characterization of an upstream open reading frame in the 5' untranslated region of PR-39, a cathelicidin antimicrobial peptide.

Cathelicidins are antimicrobial peptides present in humans, and laboratory and domestic animals. These peptides are essential components of innate and acquired immune responses; however, little is known about cathelicidin gene regulation. To investigate the regulatory mechanisms of cathelicidin gene expression, we characterized the promoter of the PR-39 gene, a multifunctional cathelicidin. Deletion analysis identified a negative regulatory element in the 5'-flanking region of the gene located in the sequence from nt (nucleotide(s)) -69 to -63. Site-directed mutagenesis indicated that ATG and its vicinity nucleotides are critical for the repressive activity of this region. A primer extension assay identified a transcription start site upstream from the negative repressor ATG and 76 nt upstream from the major open reading frame (ORF). RT-PCR and 3'-RACE further demonstrated that cDNA of PR-39 and the cathelicidin porcine myeloid antimicrobal peptide (PMAP)-23, which share prepro sequence identity, start from nt -79. Sequencing of this region showed that the 5' untranslated region (UTR) of the gene contains an upstream translation start site and an upstream ORF that functions as a repressor of the PR-39 gene. These findings indicate extensive regulation of the PR-39 gene and suggest a mechanism for the tissue-specific and age-dependent repression of this cathelicidin gene.

5' Untranslated Regions↗

PubMed2002

Astrocyte inactivation of the pRb pathway predisposes mice to malignant astrocytoma development that is accelerated by PTEN mutation.

We have inactivated pRb, p107, and p130 in astrocytes by transgenic expression of T(121) (a truncated SV40 T antigen) under the GFAP promoter. Founder mice died perinatally with extensive expansion of neural precursor and anaplastic astrocyte populations. In astrocytes, aberrant proliferation and extensive apoptosis were induced. Using a conditional allele of T(121), early lethality was circumvented, and adult mice developed high-grade astrocytoma, in which regions of decreased apoptosis expressed activated Akt. Indeed, astrocytoma development was accelerated in a PTEN(+/-), but not p53(+/-), background. These studies establish a highly penetrant preclinical model for astrocytoma based on events observed in the human disease and further provide insight into the role of PTEN mutation in astrocytoma progression.

Aging↗

PubMed2002

Modifications of the Ca2+ release mechanisms of mouse oocytes by fertilization and by sperm factor.

A cytosolic factor from sperm (SF) is thought to be responsible for the generation of intracellular calcium oscillations ([Ca2+]i) associated with fertilization in mammalian oocytes. Whether or not mouse oocytes injected with SF exhibit modifications of their Ca2+ release mechanisms similar to those observed in fertilized oocytes is not known and this was investigated here by injecting porcine SF (pSF). First, pSF-activated oocytes injected with CaCl2 showed persistent sensitization of the Ca2+-induced Ca2+ release mechanism, but this sensitization was absent in SrCl2-activated oocytes. Second, pSF-injected oocytes re-initiated oscillations when fused with untreated oocytes, although the Ca2+ responses were short-lived compared to those initiated by fertilization. Likewise, in the presence of colcemid, pSF-initiated oscillations were prolonged but ceased in advance of those in fertilized zygotes. Also, pronuclear envelope breakdown induced by okadaic acid was not associated with Ca2+ release in pSF-generated zygotes, whereas it was observed in fertilized zygotes. Finally, roscovitine, an inhibitor of maturation promoting factor, blocked pSF-induced [Ca2+]i oscillations. Together, these results show that pSF-induced [Ca2+]i responses exhibit properties similar to those triggered by the sperm, although the SF's Ca2+ active component(s) may be less stable or more susceptible to degradation, resulting in shorter modification of the oocyte's Ca2+ release mechanisms.

Animals↗

PubMed2002

Porcine sperm factor supports activation and development of bovine nuclear transfer embryos.

A study was undertaken to determine whether injection of porcine sperm factors (pSF), which trigger oscillations in intracellular calcium concentration ([Ca(2+)](i)) in mammalian oocytes, could be used to activate bovine oocytes during nuclear transfer. To date, only combined treatments that induce a monotonic rise in [Ca(2+)](i) and inhibit either phosphorylation or protein synthesis have been utilized in nuclear transfer. Several doses of pSF were tested. Injection of 5 mg/ml pSF triggered [Ca(2+)](i) oscillations that resembled those associated with fertilization with respect to amplitude and periodicity, and as a result, a high percentage of oocytes underwent activation. Furthermore, this concentration of pSF supported in vitro and in vivo development up to 60-90 days of gestation, comparable to development in control nuclear transfer embryos. Nevertheless, neither activation procedure supported development as well as did fertilization. The effectiveness of pSF as an activating agent in bovine oocytes may have been compromised because pSF was unable to support oscillations past 3-5 h postinjection and a second injection was necessary to extend the [Ca(2+)](i) oscillations. Likewise, a single injection of pSF failed to trigger downregulation of the inositol 1,4,5-trisphosphate receptor 1 subtype, whereas a second injection downregulated the receptor in a manner similar to that seen in fertilized oocytes. These results demonstrate that soluble factor(s) from porcine sperm can support early development in bovine nuclear transfer embryos; however, the efficacy may be limited because of the premature cessation of the induced oscillations.

Animals↗

PubMed2002

The role of quantitative (18)F-FDG PET studies for the differentiation of malignant and benign bone lesions.

UNLABELLED: The role of quantitative (18)F-FDG PET studies for the differentiation of benign and malignant bone lesions is still an open question. METHODS: Our evaluation included 83 patients with 37 histologically proven malignancies and 46 benign lesions. Thirty-five of the 46 benign lesions were histologically confirmed. The (18)F-FDG studies were accomplished as a dynamic series for 60 min. Evaluation of the (18)F-FDG kinetics was performed using the following parameters: standardized uptake value (SUV), global influx (Ki), computation of the transport constants K1-k4 with consideration of the distribution volume (VB) according to a 2-tissue-compartment model, fractal dimension based on the box-counting procedure (parameter for the inhomogeneity of the tumors). RESULTS: The mean SUV, the vascular fraction VB, K1, and k3 were higher in malignant tumors compared with benign lesions (t test; P < 0.05). Although the (18)F-FDG SUV was helpful to differentiate benign and malignant tumors, there was some overlap, which limited the diagnostic accuracy. On the basis of the discriminant analysis, the SUV alone showed a sensitivity of only 54.05%, a specificity of 91.30%, and a diagnostic accuracy of 74.70%. The fractal dimension was superior and showed a sensitivity of 71.88%, a specificity of 81.58%, and an accuracy of 77.14%. The combination of SUV, fractal dimension, VB, K1-k4, and Ki revealed the best results with a sensitivity of 75.86%, a specificity of 97.22%, and an accuracy of 87.69%. Bayesian analysis showed true-positive results at the level of 0.8 for a low prevalence of disease (0.235) if the full kinetic data were used in the evaluation. CONCLUSION: (18)F-FDG PET has a high specificity for the exclusion of a malignant bone tumor. Evaluation of the full (18)F-FDG kinetics and the application of discriminant analysis are required and can be used prospectively to classify a bone lesion as malignant or benign.

Bayes Theorem↗

PubMed2002

[Coronary artery bypass grafting without cardiopulmonary bypass: report of 6 cases].

OBJECTIVE: To explore the approach for coronary artery bypass grafting without cardiopulmonary bypass and assess the feasibility and safety of this procedure. METHODS: The clinical data of 6 patients receiving coronary artery bypass grafting without cardiopulmonary bypass were reviewed. RESULTS: No death occurred in these patients during the operation and the follow-up visit for 3 to 12 months found no incidence of angina pectoris, and the life quality of the patient was obviously improved. CONCLUSION: Coronary artery bypass grafting without cardiopulmonary bypass is applicable in patients with multiple coronary artery disease, especially those at high risk for cardiopulmonary bypass procedures.

Aged↗

PubMedDate not supplied

Kinetics of cold-set diffusion-limited aggregations of denatured whey protein isolate colloids.

The CaCl2-induced cold-set aggregation kinetics of the denatured whey protein isolate (WPI) colloids has been investigated under dilute diffusion-limited cluster aggregation (DLCA) conditions, using small-angle light scattering. In particular, the structure factor, the scattered intensity at zero angle and the average radius of gyration have been measured for the aggregating system as a function of time. It is found that the fractal dimension of the clusters is df= 1.85, in the range typical of clusters aggregated under DLCA conditions. The aggregation kinetics in this transition region can be described by a power law relation in the initial stage of the aggregation, but the exponent of the power law is equal to 0.7, i.e., significantly larger than 1/df= 0.54, which is the typical value of the DLCA kinetics. Since it is found that the average gyration radius of the clusters has reached a value of 80 microm, leading to a cumulative volume fraction of clusters equal to 0.25, it is legitimate to expect that the process is in the region of transition from aggregation to gelation. This confirmed by the fact that, at the later stage of the aggregation, the growth of the average cluster size further accelerates with time and eventually becomes explosive, leading to gelation. The observed aggregation kinetics has been compared with that reported in the literature from DLCA Monte Carlo simulations, and a good agreement has been found with the data corresponding to the transition region from aggregation to gelation. Numerical simulations using the Smoluchowski kinetic model have also been carried out in order to support the experimental findings.

Biocompatible Materials↗