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Biomedical subjects

X Guo

Publications and source records attributed to X Guo.

At least 343 records · Page 19Linked to original sources

Phosphatidylinositol 3-kinase in bovine photoreceptor rod outer segments.

PURPOSE: Phosphatidylinositol 3-kinase (PI 3-kinase) plays important roles in mitogenesis, vesicular trafficking, actin rearrangement, and prevention of apoptotic cell death in nonocular tissues. This investigation looked for whether PI 3-kinase is present in bovine rod photoreceptors and if light has any effect on its activity. METHODS: Bleached (BROS) and dark-adapted (DROS) rod outer segments were prepared from frozen bovine retinas and immunoblotted or immunoprecipitated with antibodies against the regulatory (p85) or catalytic (p110) subunits of PI 3-kinase. Kinase activity was measured in the immunoprecipitates and the reaction products were identified by high-performance liquid chromatography (HPLC). The amount of PI 3-kinase in membrane and cytosol fractions was determined by densitometry of immunoblots. RESULTS: Immunoblot analysis showed the presence of 85 and 110 kDa proteins in ROS. PI 3-kinase immunoprecipitated by anti-p85 antibody converted PI to PI-3-P and PI-4-P to PI-3,4-P2, as determined by HPLC analysis of the deacylated products. The PI 3-kinase activity in these ROS preparations was sensitive to wortmannin, a potent inhibitor of PI 3-kinase, at low concentrations (IC50 3 nM). Immunoprecipitates (IPs) showed activity twice as high in BROS as in DROS. The IPs of ROS membranes but not cytosol maintained the light-dark difference. However, measurement of anti-p85 and anti-p110 immunoreactivities on western blots of ROS, ROS membranes, and ROS cytosol did not show any light-dark differences. CONCLUSIONS: PI 3-kinase is present in bovine rod outer segments and its activity appears to be greater in light-adapted retinas.

Animals↗

Change of foot size with weightbearing. A study of 2829 children 3 to 18 years of age.

The change of foot length and width with age has been reported in a few anthropometric studies in the literature. However, the relationship with body height rarely is reported, and the dynamic effect of weightbearing on foot size has not been documented. In this series, 2829 children 3 to 18 years of age of equal gender distribution were included in the study. The foot length and width on weightbearing and nonweightbearing were measured with a special precision electronic caliper. The foot length and width were found to increase linearly from the age of 3 years until 12 years in girls and 15 years in boys. This was followed by a phase during which the increase plateaued. The foot length and width increased significantly on weightbearing at all ages in both genders with a mean of 2.1 to 4.4 mm or 3.1% to 4.8%, respectively. The foot length and width also were found to correlate significantly with the body height in both genders, with a correlation coefficient of 0.96 to 0.98. No significant differences were found between the sizes of the dominant and nondominant foot in either gender.

Adolescent↗

[Effects of subretinal fluid containing proteins with different molecular weights on the proliferative vitreoretinopathy].

The stimulating effects of subretinal fluid (SRF) containing different molecular weight proteins which were isolated by fast performance liquid chromatography (FPLC) and ultrafiltration on the proliferative vitreoretinopathy (PVR) were investigated. The results showed that (1) the PVR was induced by 10-30 kD proteins injecting vitreously, and the forming duration of PVR was about two weeks; (2) the location of inducing PVR was mainly on medullary ray area in the rabbit eye; (3) In the SRF there were growth factors which were mainly those proteins with molecular weight between 10 kD and 30 kD.

Adult↗

[Clinical application of myocardial perfusion imaging with 99mTc-MIBI SPECT in the diagnosis of coronary artery disease].

Myocardial perfusion imaging with 99mTc-MIBI was studied in 80 patients with coronary artery disease (CAD), 29 cases of normal controls and 37 cases of noncoronary heart diseases. The perfusion imaging was analysed both by qualitative and quantitative methods. The results revealed that the diagnostic sensitivity, specificity and accuracy for CAD were 91.5%, 83.8%, and 88.5% respectively by qualitativ method, and were 88.1%, 94.6% and 90.6% respectively by quantitative method. The diagnostic sensitivity of electrocardiogram (ECG) was 69.5%. Fourteen cases with the narrowing of coronary artery > 50% shown by coronary angiography also had abnormal myocardial perfusion imaging. It suggests that analysis of perfusion imaging by quantitative method in detecting CAD is superior than qualitative method and ECG.

Adult↗

[The effect of root canal irrigation and variance of apical foramen pressure by use of casing aspiration needle system in vitro].

The authors introduced casing aspiration needle system (CANS) to irrigating root canal. The effect of root irrigation on removing canal debris and the variance of apical foramen pressure on extracted teeth were compared between the conventional irrigation and CANS irrigation. The results indicated that the effect of root canal irrigation by CANS was much better than the conventional method (P < 0.01). CANS irrigation didn't produce pressure to apical foramen during root canal irrigation. However, in the conventional method, with irrigation needle inserted deeper and deeper in root cancal the pressure of apical foramen gets higher and higher.

Humans↗

Characterization of a replication-incompetent adenovirus type 5 mutant deleted for the preterminal protein gene.

An adenovirus type 5 mutant deleted for the preterminal protein (pTP) gene was constructed using cell lines that express pTP. The pTP deletion mutant virus is incapable of replicating in the absence of complementation and does not express detectable levels of viral mRNAs that are expressed only after the onset of replication. Accumulation of early-region mRNAs, including that for E1A, exhibits a lag relative to that observed from the wild-type virus. However, E1A mRNA accumulation attains a steady-state level similar to the level of expression during the early phase of infection with the wild-type virus. In 293-pTP cells (human embryonic kidney cells that express pTP in addition to high levels of adenovirus E1A and E1B proteins), the pTP deletion mutant virus replicates efficiently and yields infectious titers within 5-fold of that of the wild-type virus. The deletion of 1.2 kb of pTP-encoding sequence increases the size of foreign DNA that can be introduced into the virus and, with an absolute block to replication, makes this virus an important tool for gene therapy.

Adenoviruses, Human↗

Differential regulation of cardiac actomyosin S-1 MgATPase by protein kinase C isozyme-specific phosphorylation of specific sites in cardiac troponin I and its phosphorylation site mutants.

The significance of site-specific phosphorylation by protein kinase C (PKC) isozymes alpha and delta and protein kinase A (PKA) of troponin I (TnI) and its phosphorylation site mutants in the regulation of Ca(2+)-stimulated MgATPase activity of reconstituted actomyosin S-1 was investigated. The genetically defined TnI mutants used were T144A, S43A/S45A, S43A/S45A/T144A (in which the PKC phosphorylation sites Thr-144 and Ser-43/Ser-45 were respectively substituted by Ala) and N32 (in which the first 32 amino acids in the NH2-terminal sequence containing Ser-23/Ser-24 were deleted). Although the PKC isozymes displayed different substrate phosphorylation kinetics, PKC-alpha phosphorylated equally well TnI wild type and all mutants, whereas N32 was a much poorer substrate for PKC-delta. Furthermore, the two PKC isozymes exhibited discrete specificities in phosphorylating distinct sites in TnI and its mutants, either as individual subunits or as components of the reconstituted troponin complex. Unlike PKC-alpha, PKC-delta favorably phosphorylated the PKA-preferred site Ser-23/Ser-24 and hence, like PKA, reduced the Ca2+ sensitivity of the reconstituted actomyosin S-1 MgATPase. In contrast, PKC-alpha preferred to phosphorylate Ser-43/Ser-45 (common sites for all isozymes) and thus reduced the maximal Ca(2+)-stimulated activity of the MgATPase. In this respect, PKC-delta, by cross-phosphorylating the PKA sites, functioned as a hybrid of PKC-alpha and PKA. The site specificities and hence functional differences between PKC-alpha and -delta were most evident at low phosphorylation (1 mol of phosphate/mol) of TnI wild type and were magnified when S43A/S45A and N32 were used as substrates. The present study has demonstrated, for the first time, that distinct functional consequences could arise from the site-selective preferences of PKC-alpha and -delta for phosphorylating a single substrate in the myocardium, i.e., TnI.

Actomyosin↗

KAI1 expression is up-regulated in early pancreatic cancer and decreased in the presence of metastases.

KAI1 is a metastasis suppressor gene for prostate cancer that is located on chromosome 11p11.2-13. Using Northern blot analysis and in situ hybridization, we studied expression of KAI1 mRNA in specimens from 14 normal pancreases and 27 primary pancreatic cancers, and then correlated the findings with the clinical and histopathological parameters of the patients. Northern blot analysis showed increased steady-state levels of KAI1 mRNA expression in 24 of 27 (89%) pancreatic cancer samples. In situ hybridization showed enhanced KAI1 mRNA levels in the pancreatic cancer cells in 82% cancer tissues. The stroma surrounding the cancer mass and normal pancreatic tissue adjacent to the cancer cells exhibited very low levels of KAI1 mRNA expression. Correlation of the mRNA levels obtained by Northern blot analysis with clinical parameters of the patients revealed that KAI1 mRNA levels were significantly higher (P < 0.01) in earlier tumor stages (I, II), compared with advanced tumor stages (III, IV) in which lymph node or distant metastases were present. Furthermore, poorly differentiated tumors had significantly higher KAI1 mRNA levels than those that were moderately or well differentiated (P < 0.05). No association between KAI1 expression and survival was found. Our results indicate that KAI1 mRNA expression is reduced in patients with advanced tumor stages. This suggests that reduction of KAI1 expression might enable pancreatic cancer cells to spread in lymph nodes and to distant organs.

Adult↗

A non-cholinergic transmitter, pituitary adenylate cyclase-activating polypeptide, utilizes a novel mechanism to evoke catecholamine secretion in rat adrenal chromaffin cells.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is the most potent non-cholinergic neurotransmitter to stimulate catecholamine secretion from rat chromaffin cells; however, the mechanism of action is not clear. We used amperometric detection of exocytosis and indo-1 monitoring of [Ca2+]i to identify PACAP actions in cultured chromaffin cells. PACAP (100 nM) required external Ca2+ to evoke secretion. However, unlike nicotine and KCl which caused immediate and relatively brief secretion, PACAP has a latency of 6.8 +/- 0.96 s to the first secretory response and secretion continued for up to 2 min. PACAP elevation of [Ca2+]i showed similar latency and often remained above base line for several minutes following a brief exposure. ZnCl2 (100 microM) selectively inhibited PACAP-stimulated secretion and [Ca2+]i with little effect on nicotine-evoked responses. Nifedipine (10 microM) had little effect on PACAP-evoked secretion but inhibited nicotine-evoked secretion by more than 80%, while omega-conotoxin (100 nM) failed to affect either agonist. PACAP-stimulated cAMP levels required 5 s to significantly increase, consistent with the latency of exocytotic and Ca2+ responses. Forskolin (10 microM) caused responses similar to PACAP. PACAP-evoked exocytosis was blocked by the protein kinase A inhibitor adenosine 3'5'-cyclic monophosphorothioate Rp-diastereomer (Rp-cAMPS). These data showed that PACAP stimulates exocytosis by a mechanism distinctly different from cholinergic transmitters that appears to involve cAMP-mediated Ca2+ influx. Differences in receptor coupling mechanisms and pharmacology of Ca2+ entry stimulated by cholinergic and peptidergic agonists support the idea that the peptidergic system maintains catecholamine secretion under conditions where the cholinergic system desensitizes or otherwise fails.

Adrenal Glands↗

Homogeneous enzyme-based binding assay for studying glycosaminoglycan interactions with macromolecules and peptides.

A simple and rapid homogeneous enzyme-based binding assay is described to study the degree of interaction between glycosaminoglycans and various macromolecules/peptides. The method is based on the homogeneous inhibition of a highly positively charged enzyme, acid deoxyribonuclease II (EC 3.1.22.1), by glycosaminoglycan polyanions, such as heparin, chondroitin 4-sulfate, and dermatan sulfate. Catalytic activity of DNase II is inhibited to nearly 100% by relatively small amounts of these glycosaminoglycan molecules. In the presence of species that bind these polyanions, the activity of the enzyme is regained in an amount proportional to the concentration of the species present. Thus, the relative binding affinities of various species with a given GAG can be assessed rapidly by comparing the concentration of the compound required to reverse the enzyme inhibition to 50% of the maximum value (ED(50) values). The feasibility of this binding assay principle is demonstrated by measuring the ED(50) values of five macromolecules: polylysine, polyarginine, protamine, low-density lipoprotein (LDL), and high-density lipoprotein (HDL), using heparins of different size, as well as chondroitin 4-sulfate and dermatan sulfate as the GAG polyanions. The applicability of the assay method is further extended to study GAG-peptide interactions. A variety of small synthetic peptides (8-13 amino acid residues) derived from the heparin-binding domains of protamine and type IV collagen are used as model peptide species. Relative GAG-binding affinities of these macromolecules/peptides are compared to previous literature values and data obtained via a new electrode-based titration method.

Binding, Competitive↗