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Biomedical subjects

Y Yao

Publications and source records attributed to Y Yao.

At least 163 records · Page 9Linked to original sources

Isolation and characterization of cDNA encoding mouse RNA polymerase II subunit RPB14.

By means of the yeast two-hybrid system using the 40-kDa subunit of mouse RNA polymerase I, mRPA40, as the bait, we isolated a mouse cDNA which encoded a protein with significant homology in amino acid sequence to the 12.5-kDa subunit of Saccharomyces cerevisiae RNA polymerase II, B12.5 (RPB11). Specific antibody raised against the recombinant protein that was derived from the cDNA reacted with a 14-kDa polypeptide in highly purified mammalian RNA polymerase II and did not react with any subunit of RNA polymerase I or III. Moreover, the antibody co-immunoprecipitated the largest subunit of mouse RNA polymerase II. These results provide biochemical evidence that the cDNA isolated, named mRPB14, encodes a specific subunit of RNA polymerase II, and indicate that the subunit organization of the enzyme is conserved between yeast and mouse. A possible role of the alpha-motif [Dequard-Chablat, M., Riva, M., Carles, C. and Sentenac, A., J. Biol. Chem. 266 (1991) 15300-15307] in the protein-protein interaction between mRPA40 and mRPB14 is also discussed.

Amino Acid Sequence↗

Adenosine A3 receptor agonists protect HL-60 and U-937 cells from apoptosis induced by A3 antagonists.

The effects of novel, selective adenosine (ADO) A3 receptor antagonists of diverse structure on cells of the human HL-60 leukemia and U-937 lymphoma cell lines were examined. Both 3-ethyl 5-benzyl 2-methyl-6-phenyl-4-phenylethynyl-1,4-(+/-)-dihydropyridine-3, 5-dicarboxylate (MRS 1191, 0.5 microM) and 6-carboxy-methyl-5, 9-dihydro-9-methyl-2-phenyl-[1,2,4]-triazolo [5,1-a][2,7]naphthyridine (L-249313, 0.5 microM) induced apoptotic cell death and expression of bak protein. Low concentrations of the A3 receptor agonist 2-chloro-N6-(3-iodobenzyl)adenosine-5'-N-methyluronamide (Cl-IB-MECA, 10 nM or 1 microM) protected against antagonist-induced cell death. At concentrations > or = 10 microM, the agonist alone produced apoptosis and bak expression in various cell lines. It is suggested that there exists a tonic low level of A3 receptor activation, possibly induced by release of endogenous adenosine, that results in cell protection.

Adenosine↗

Expression of scatter factor and c-met receptor in benign and malignant breast tissue.

BACKGROUND: Scatter factor (SF), also known as hepatocyte growth factor, is an angiogenic cytokine that stimulates epithelial cell motility and invasion. Its receptor is a transmembrane tyrosine kinase encoded by the c-met protooncogene. Several prior experimental and clinical studies have suggested that SF might play a role in the development and progression of breast carcinoma. To investigate the possible involvement of SF and c-met in the evolution of breast carcinoma, the authors studied their expression in sections of human breast tissue. METHODS: A variety of paraffin embedded tissue specimens (of normal breast tissue tissue, benign hyperplasia, ductal carcinoma-in-situ [DCIS], and invasive ductal carcinoma) from 125 patients were immunoperoxidase-stained using specific antisera against SF and c-met. The staining intensities of epithelial mammary cells were scored semiquantitatively, and the staining scores were analyzed as a function of tissue type. In addition, in situ hybridization to detect SF mRNA was performed for a small number of cancer sections. RESULTS: Specific SF staining was observed in tumor cells, normal cell types (epithelium and vascular smooth muscle), and acellular stroma, whereas c- met staining was observed in tumor cells and normal cell types but not in stroma. Analysis of the staining scores of epithelial mammary cells revealed several patterns: (1) SF and c-met staining scores each increased in the following order: normal breast/benign hyperplasias (lowest) --> DCIS (higher) --> invasive carcinoma (highest); (2) normal-appearing mammary ducts and lobules in invasive cancer sections showed less SF and c-met staining than tumor cells in the same specimens but more staining than normal ducts and lobules in sections of normal breast tissue and benign hyperplasia; (3) within the DCIS and invasive cancer groups, SF and c-met staining scores were correlated; and (4) among 40 consecutive cases of DCIS, higher levels of SF and c-met staining showed a trend toward association with other features suggestive of aggressive tumor biology (comedo histology, high nuclear grade, p53 positivity, and bcl-2 negativity). In situ hybridization analysis revealed that the same cell types that expressed SF protein (including tumor cells) also expressed SF mRNA transcripts. CONCLUSIONS: SF and c-met are overexpressed in breast carcinoma as compared with benign breast tissue, and they tend to be coexpressed in cancerous tissue. These findings are consistent with the idea that the SF:c-met ligand:receptor pair may have a role in breast carcinoma progression.

Breast↗

Both the isomerase and chaperone activities of protein disulfide isomerase are required for the reactivation of reduced and denatured acidic phospholipase A2.

The spontaneous reactivation yield of acidic phospholipase A2 (APLA2), a protein containing seven disulfide bonds, after reduction and denaturation in guanidine hydrochloride is very low. Protein disulfide isomerase (PDI) markedly increases the reactivation yield and prevents the aggregation of APLA2 during refolding in a redox buffer containing GSH and GSSG. S-methylated PDI (mPDI), with no isomerase but as nearly full chaperone activity as native PDI, has no effect on either the reactivation or aggregation of APLA2. However, the simultaneous presence of PDI and mPDI in molar ratios to APLA2 of 0.1 and 0.9 respectively fully reactivates the denatured enzyme, as does PDI alone at a ratio of 1. At ratios of 0.1 and 0.15 respectively, they completely suppress APLA2 aggregation, as does PDI alone at a ratio of 0.25. Moreover, delayed addition of PDI to the refolding buffer greatly diminished the reactivation yield of APLA2, but this deteriorating effect can be alleviated markedly by the presence of mPDI in the refolding buffer. Without GSSG, mPDI prevents the aggregation of APLA2 during refolding. It is proposed that the in vitro action of PDI as a foldase consists of both isomerase and chaperone activities, and the latter activity can be fully replaced by mPDI.

Agkistrodon↗

Alport syndrome with neurofibromatosis type-I: a case report.

We report a 9-year-old boy with repeated fractures of the tibia from age 6 months and microscopic hematuria from age 2 years. His maternal family has a history of nephritis and his paternal family has neurofibromatosis type-I (NF-I). The boy's renal biopsy revealed an irregular attenuation and splitting of the glomerular basement membrane. The skin biopsy was stained with monoclonal antibody against the alpha 5 chain of type IV collagen; the epidermal basement membrane was negative in the boy and segmentally positive in the boy's mother. We conclude that the patient inherited Alport syndrome from his mother and NF-I from his father. We postulate this was a chance association and that this case does not suggest any relationship between the two diseases.

Child↗

Calcium current activated by depletion of calcium stores in Xenopus oocytes.

Ca(2+) currents activated by depletion of Ca(2+) stores in Xenopus oocytes were studied with a two-electrode voltage clamp. Buffering of cytosolic Ca(2+) with EGTA and MeBAPTA abolished I(Cl(Ca)) and unmasked a current in oocytes that was activated by InsP(3) or ionomycin in minutes and by thapsigargin or the chelators themselves over hours. At -60 mV in 10 mM extracellular CaCl(2), the current was typically around -90 or -160 nA in oocytes loaded with EGTA or MeBAPTA, respectively. This current was judged to be a Ca(2+)-selective current for the following reasons: (a) it was inwardly rectifying and reversed at membrane potentials usually more positive than +40 mV; (b) it was dependent on extracellular [CaCl(2)] with K(m) = 11.5 mM; (c) it was highly selective for Ca(2+) against monovalent cations Na(+) and K(+), because replacing Na(+) and K(+) by N-methyl-d-glucammonium did not reduce the amplitude or voltage dependence of the current significantly; and (d) Ca(2+), Sr(2+), and Ba(2+) currents had similar instantaneous conductances, but Sr(2+) and Ba(2+) currents appeared to inactivate more strongly than Ca(2+). This Ca(2+) current was blocked by metal ions with the following potency sequence: Mg(2+) << Ni(2+) approximately Co(2+) approximately Mn(2+) < Cd(2+) << Zn(2+) << La(3+). It was also inhibited by niflumic acid, which is commonly used to block I(Cl(Ca)). PMA partially inhibited the Ca(2+) current, and this effect was mostly abolished by calphostin C, indicating that the Ca(2+) current is sensitive to protein kinase C. These results are the first detailed electrophysiological characterization of depletion-activated Ca(2+) current in nondialyzed cells. Because exogenous molecules and channels are easy to introduce into oocytes and the distortions in measuring I(Cl(Ca)) can now be bypassed, oocytes are now a superior system in which to analyze the activation mechanisms of capacitative Ca(2+) influx.

Animals↗

A wavelet-based multiresolution regularized least squares reconstruction approach for optical tomography.

In this paper, we present a wavelet-based multigrid approach to solve the perturbation equation encountered in optical tomography. With this scheme, the unknown image, the data, as well as the weight matrix are all represented by wavelet expansions, thus yielding a multiresolution representation of the original perturbation equation in the wavelet domain. This transformed equation is then solved using a multigrid scheme, by which an increasing portion of wavelet coefficients of the unknown image are solved in successive approximations. One can also quickly identify regions of interest (ROI's) from a coarse level reconstruction and restrict the reconstruction in the following fine resolutions to those regions. At each resolution level a regularized least squares solution is obtained using the conjugate gradient descent method. This approach has been applied to continuous wave data calculated based on the diffusion approximation of several two-dimensional (2-D) test media. Compared to a previously reported one grid algorithm, the multigrid method requires substantially shorter computation time under the same reconstruction quality criterion.

Algorithms↗

Frequency-domain optical imaging of absorption and scattering distributions by a Born iterative method.

We presents a Born; iterative method, for reconstructing optical properties of turbid media by means of frequency-domain data. The approach is based on iterative solution of a linear perturbation equation, which is derived from the integral from of the Helmholtz wave equation for photon-density waves in each iteration the total field and the associated weight matrix are recalculated based on the previous reconstructed image. We then obtain a new estimate by solving the updated perturbation equation. The forward solution, also based on a Helmholtz equation, is obtained by a multigrid finite difference method. The inversion is carried out through a Tikhonov regularized optimization process by the conjugate gradient descent method. Using this method, we first reconstruct the distribution of the complex wave numbers in a test medium, from which the absorption and the scattering distributions are then derived. Simulation results with two-dimensional test media have shown that this method can yield quantitatively (in terms of coefficient valued) as well as qualitatively (in terms of object location and shape) accurate reconstructions of absorption and scattering distributions in cases in which the first-order Born approximation cannot work well. Both full-angle and limited-angle measurement schemes have been simulated to examine the effect of the location of detectors and sources. The robustness of the algorithm to noise has also been evaluated.

Absorption↗

Iterative total least-squares image reconstruction algorithm for optical tomography by the conjugate gradient method.

We present an iterative total least-squares algorithm for computing images of the interior structure of highly scattering media by using the conjugate gradient method. For imaging the dense scattering media in optical tomography, a perturbation approach has been described previously [Y. Wang et al., Proc. SPIE 1641, 58 (1992); R. L. Barbour et al., in Medical Optical Tomography: Functional Imaging and Monitoring (Society of Photo-Optical Instrumentation Engineers, Bellingham, Wash., 1993), pp. 87-120], which solves a perturbation equation of the form W delta x = delta I. In order to solve this equation, least-squares or regularized least-squares solvers have been used in the past to determine best fits to the measurement data delta I while assuming that the operator matrix W is accurate. In practice, errors also occur in the operator matrix. Here we propose an iterative total least-squares (ITLS) method that minimizes the errors in both weights and detector readings. Theoretically, the total least-squares (TLS) solution is given by the singular vector of the matrix [W/ delta I] associated with the smallest singular value. The proposed ITLS method obtains this solution by using a conjugate gradient method that is particularly suitable for very large matrices. Simulation results have shown that the TLS method can yield a significantly more accurate result than the least-squares method.

Algorithms↗

[Effect of catecholaminergic agonists and antagonists on adaptive gastric mucosal protection in rats].

The gastric mucosal damage caused by 70% ethanol significantly decreased in rats pretreated with chronic mild restraint stress (CMRS, P < 0.001). This adaptive mucosal protection induced by CMRS was absent in sympathectomized rats, while the administration of isoprenaline or dopamine could partially, and norepinephrine could not, restore the protection. In rats with rntact sympathetic nerve, preinjection of haloperidol or propranolol inhibited CMRS-induced protection which, however, was not affected by phentolamine. No change in plasma somatostatin level was observed either in stress or sympathectomized rats alone or in stress plus sympathectomized rats. The results suggest that the sympathetic nerve or its mimetic agent ang this is are involved in the adaptive protection of the gastric mucosa possibly mediated through beta and dopamine receptors.

Adaptation, Physiological↗

[The relationship between abnormalities of cell-mediated immunity and gut origin endotoxemia in a rat model of thermal injury].

This study was conducted to determine the relationship between abnormalities of cell-mediated immunity and gut-derived endotoxemia in rats following burns. Animals were subjected to a 40% full-thickness scald injury, and randomly divided into control and selective decontamination of the digestive tract (SDD) treated groups. It was found that thermal injury resulted in marked reductions in splenocyte proliferative response to concanavalin A or phytohemagglutinin, interleukin 2 (IL-2) production, and T helper/suppressor (Th/Ts) cells ratio. Prophylactic treatment with SDD significantly reduced the incidence of endotoxemia, prevented suppressive mitogenic response and inadequacy in IL-2 production (P < 0.05-0.01), but did not affect the abnormal ratio of Th/Ts in blood (P > 0.05). We conclude that bacteria/endotoxin translocation from the gastrointestinal tract appears to be involved in cellular immune dysfunction after thermal injury. Pretreatment with SDD might attenuate systemic immunosuppression by preventing translocation events.

Animals↗

[Serum neopterin levels after extensive burns and their relationship to endotoxemia and sepsis].

The present study was performed to determine the kinetics of serum neopterin levels after major thermal injury and their relationship to endotoxemia and sepsis. This prospective study included 35 patients (32 males and three females) with total burn surface area (TBSA) greater than 30% (30%-98%), and 22 healthy volunteers who served as normal controls. The results showed that neopterin levels increased in most patients on day 3 postburn, but they were not significantly correlated with the extent of the burn surface (P > 0.05). Patients with endotoxemia had much higher neopterin values than those who showed no endotoxemia from the second week onward (P < 0.05-0.01), and circulating endotoxin and neopterin levels were positively correlated in patients who developed endotoxemia on day 14 (r = 0.368, P < 0.05) and day 21 (r = 0.439, P < 0.01) after major burns. Moreover, a high serum neopterin level was found in patients with sepsis (n = 15), and the marked elevation persisted throughout the observation period. The difference between septic and non-septic patients (n = 20) became significant on 14 and 28 days postburn. These data suggest that extensive burns can lead to an elevation of serum neopterin independent of TBSA. The endotoxin release in the circulation may be involved in the continuous formation of neopterin during the late postburn stage. In addition, the presence of a constant high neopterin level is associated with a critical event in the development of burn sepsis.

Adolescent↗

[Effect of recombinant bactericidal/permeability-increasing protein on pulmonary cytokine mRNA expression in rats following hemorrhage and resuscitation].

To determine the possible mechanisms underlying beneficial effect of recombinant bactericidal/permeability-increasing protein (rBPI) on acute lung injury response to blood loss, we used reverse transcription polymerase chain reaction to measure pulmonary tumor necrosis factor (TNF), interleukin 6 (IL-6) mRNA expression in a rat model of prolonged hemorrhagic shock (4.00 kPa, 180 min) followed by adequate resuscitation. The results showed that systemic plasma endotoxin concentrations elevated rapidly after a 180-min hemorrhagic insult (P < 0.05), and TNF, IL-6 mRNA expression in the lung were significantly increased at 2, 8 hours after resuscitation respectively. However, treatment with rBPI resulted in almost neutralization of plasma endotoxin values, remarkable reduction of TNF, IL-6 mRNA levels following hemorrhage/resuscitation. Also, it was found that rBPI administration markedly blunted the increase in pulmonary Evans blue dye extravasation, concomitant with a significant decrease in lung myeloperoxidase activity compared with the control group (P < 0.05-0.01). These data suggest that local proinflammatory cytokine mRNA expression associated with gut origin endotoxemia may be an important mechanism contributing to the development of hemorrhage-induced lung injury. Treatment with rBPI is effective in inhibiting marked TNF, IL-6 mRNA expression and ameliorating acute lung injury secondary to severe hemorrhagic shock.

Animals↗

[Penetration of ciprofloxacin and cefoperazone into human pancreas].

Major pancreatic infection is responsible for more than 80% of deaths in patients with acute pancreatitis. Therefore, the role of antimicrobial drugs in the prevention and treatment of secondary parcreatic infection is very important. The choice of antimicrobial drugs must be based upon the ability of the drug to exceed the therapeutic concentration in pancreas for the common pathogens. The penetration of ciprofloxacin and cefoperazone into pancreas was investigated in ten patients who had undergone pancreatoduodenectomy. The pancreatic juice was temporarily diverted to the exterior via a panoreatic duct catheter. The pancreatic tissue was obtained intraoperatively and pancreatic juice was drained postoperatively. The antimicrobial drug concentrations were determined by high-performance liquid chromatography. The concentrations of ciprofloxacin and cefoperazone in pancreatic juice were 44% and 17%, respectively, of those in serum, and exceeded the in vitro concentration (MIC-90) for most bacteria associated with pancreatic infections. The result indicates that ciprofloxacin and cefoprazone appear to be appropriate for both prophylaxis and therapy of secondary pancreatic infections.

Adult↗

[The changes of glutamate receptor and free Ca2+i in hypoxic-ischemic cerebral injury: experimental study].

This experiment was designed to explore the pathogenesis in hypoxic-ischemic encephalopathy (HIE). Sixteen newborn pigs were divided into two groups: (Group A) normal control and (Group B) HIE 24 hours. The glutamate receptor (Glu R) in forebrain crude synaptic membrane (SPM) and free Ca2+i in RBC were tested respectively. The results revealed that the binding sites (Bmax) of Glu R in Group B was much lower than that in Group A, but the affinity (Kd) showed no statistic difference between Group A and Group B. In addition, free Ca2+i of RBC in Group B was much higher than that in Group A. This study demonstrates that the changes of Glu R and Ca2+i are involved in the pathogenesis of hypoxic-ischemic cerebral injury in newborn animals.

Animals↗

[Observation on the changes of lipid peroxidation in neonates with sclerema].

Thirty-six hospitalized newborn infants with sclerema (case group), which were divided into mild group(n = 18) and severe group(n = 18), and 28 normal neonates (normal control group) were selected to measure the changes of blood lipid peroxidation (LPO) and superoxide dismutase(SOD) so as to study the role of free radicals and lipid peroxide injury the pathogenesis of the entity. The results showed that the plasma LPO contents were significantly increased and SOD were significantly decreased in the case group when compared with those in the normal group (P < 0.01). The plasma LPO was significantly higher and SOD activity was significantly lower in the severe group than those in the mild group (P < 0.01). The study indicates that the free radicals produced in sclerema neonatorum are great in amount and suggests that free radicals possibly participate in the pathogenesis of the disease.

Erythrocytes↗

[Effect of loss of consciousness induced by repeated lower body negative pressure on blood-brain barrier permeability in rats].

To investigate the effect of loss of consciousness induced by repeated +Gz on the brain and its mechanism, changes of blood-brain barrier (BBB) permeability and brain water content after repeated lower body negative pressure (LBNP) induced loss of consciousness were observed in 30 rats. Rats were anesthetized and exposed to LBNP (-4.0 kPa). The pressure rapidly returned to control level 2 min after EEG became isoelectric. The BBB permeability was measured using lanthanum trace labeling with electron microscopy and brain water content was determined after one or three LBNP exposures. Most tight junctions among endothelial cells opened through which the lanthanum entered into the neurons and extra cellular space and water content in brain tissues increased significantly (P < 0.05) 1 h after 3 LBNP exposures. It demonstrated that 3 exposures to LBNP induced loss of consciousness can result in an increase of BBB permeability in rats, which may play an important role in the development of G-induced brain edema.

Aerospace Medicine↗