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Biomedical subjects

Z Liu

Publications and source records attributed to Z Liu.

At least 1,243 records · Page 69Linked to original sources

High-affinity binding of progesterone to the plasma membrane of Xenopus oocytes: characteristics of binding and hormonal and developmental control.

Progesterone induces maturation of the amphibian oocyte through its action on the plasma membrane. However, whether or not this action requires high-affinity binding to a specific receptor is unclear. In this study, the binding activity of progesterone was characterized in plasma membranes from whole ovaries or defolliculated oocytes of Xenopus laevis. Membrane-bound, radiolabeled progesterone was isolated by filtration of membrane suspensions and quantified by liquid scintillation. The association of progesterone to membrane preparations reached equilibrium within 15 min. Progesterone binding activity was directly proportional to the sample concentration, was significantly reduced by trypsin digestion, and was pH-dependent and temperature-sensitive. Also, binding activity was observed in membrane preparations from whole ovaries and defolliculated oocytes but not in those from somatic cells, indicating that progesterone binding is restricted to the oocyte membrane. Scatchard analysis indicated a single class of high-affinity (average KD, 10(-9) M), low-capacity (average concentration, 10(-12) mol/mg protein) binding sites for progesterone in all oocyte membrane preparations tested. Progesterone binding activity was also detected in preparations from albino frog ovaries, indicating that the binding activity is not an artifact of melanin contamination. Competition studies showed the following order of affinities: progesterone > pregnenolone > 17 alpha, 20 beta, 21-trihydroxy-4-pregnen-3-one > 11-deoxycorticosterone > 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one > 11-deoxycortisol > estradiol > R5020 > corticosterone > aldosterone > cortisol > androstenedione > ouabain.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Characterization of skeletal muscle actin labeled with the triplet probe erythrosin-5-iodoacetamide.

We have labeled rabbit skeletal muscle actin with the triplet probe erythrosin-5-iodoacetamide and characterized the labeled protein. Labeling decreased the critical concentration and lowered the intrinsic viscosity of F-actin filaments; labeled filaments were motile in an in vitro motility assay but were less effective than unlabeled F-actin in activating myosin S1 ATPase activity. In unpolymerized globular actin (G-actin), both the prompt and delayed luminescence were red-shifted from the spectra of the free dye in solution and the fluorescence anisotropy of the label was high (0.356); filament formation red shifted all excitation and emission spectra and increased the fluorescence anisotropy to 0.370. The erythrosin phosphorescence decay was at least biexponential in G-actin with an average lifetime of 99 microseconds while in F-actin the decay was approximately monoexponential with a lifetime of 278 microseconds. These results suggest that the erythrosin dye was bound at the interface between two actin monomers along the two-start helix. The steady-state phosphorescence anisotropy of F-actin was 0.087 at 20 degrees C and the anisotropy increased to approximately 0.16 in immobilized filaments. The phosphorescence anisotropy was also sensitive to binding the physiological ligands phalloidin, cytochalasin B and tropomyosin. This study lays a firm foundation for the use of this triplet probe to study the large-scale molecular dynamics of F-actin.

Actins↗

The cellular source of interleukin-6 during Listeria infection.

The cellular source of interleukin-6 (IL-6) during infection of mice with Listeria monocytogenes was investigated both in vitro and in vivo. Peritoneal cells taken at intervals from infected mice and cultured in vitro without added stimulus produced high titers of IL-6 peaking 2 days postinfection in a time course similar to that observed in vivo. Adherent cells with the morphology of macrophages were a major source of this IL-6. Spleen cells similarly harvested at intervals and cultured with heat-killed Listeria or heat-killed Brucella organisms as specific and nonspecific stimuli, respectively, showed two distinct IL-6 responses: (i) an early-phase response up to 5 days after infection when IL-6 production was elicited by either a specific or nonspecific stimulus, and when depletion of T cells had no effect, and (ii) a later response 7 to 10 days after infection when very high levels of IL-6 were produced in response to a specific stimulus. This response was lost when T cells were depleted in vitro or in vivo or in spleen cell cultures from mice with severe combined immunodeficiency. However, studies in vivo failed to show an important role for T cells governing serum IL-6. We conclude that most of IL-6 detected in vivo is produced by nonlymphocytes. Whether IL-6 produced by T lymphocytes in local foci of infection has any role in resolution of that infection is unknown.

Animals↗

Selective sequestration of nitric oxide by subcellular components of vascular smooth muscle and platelets: relationship to nitric oxide stimulation of the soluble guanylyl cyclase.

Sequestration of nitric oxide (NO) by subcellular fractions isolated from bovine pulmonary arterial medial layer (BPA) and rabbit platelets (RP) was studied utilizing a novel chemiluminescence--headspace gas technique. Sequestration in all fractions was similarly rapid (5 min) and remained constant for at least 30 min. When incubated with 108 pmol of NO, the BPA mitochondrial, microsomal, and nuclear fractions sequestered 22.8 +/- 1.9, 20.5 +/- 2.2 and 15.2 +/- 3.6% of the NO, respectively (n = 14). However, significantly more of the 108 pmol of NO, 36.8 +/- 2.8 and 32.9 +/- 3.6%, respectively, was sequestered by the BPA homogenate (about 2 mg protein/mL) and BPA cytosolic fraction (about 1 mg protein/mL) (n = 19). Also, RP cytosolic fraction (about 3 mg protein/mL) sequestered a greater amount of NO than any BPA fraction when incubated with 108 pmol of NO (83.0 +/- 1.0%; n = 3). Analysis of the binding data obtained for the BPA homogenate and cytosolic fraction was consistent with the existence of two binding sites, one site with a Kd of approximately 100 nM and another with a Kd of approximately 1 microM. Both the BPA homogenate fraction and the cytosolic fraction as well as the RP cytosolic fraction were shown to have soluble guanylyl cyclase activity. The nitrovasodilator sodium nitroprusside (SNP) caused a concentration-dependent increase in the activity of this enzyme in all these fractions. Maximum stimulations caused by 1 mM SNP in BPA homogenate fraction, BPA cytosolic fraction, and RP cytosolic fraction were equivalent to 2-, 4- and 3-fold increases in catalytic activity, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Mechanisms of adaptation of glucose transporters to changes in the oxidative chain of muscle and fat cells.

Cells in which glucose uptake is rate limiting respond to hypoxic insults with an increase in glucose transport activity. To understand the underlying cellular mechanisms involved in this adaptive response, the effects of an uncoupler of oxidative phosphorylation, 2,4-dinitrophenol (DNP), and of an inhibitor of the electron transport chain, rotenone, were compared with the effect of hypoxia in L6 muscle cells and 3T3-L1 adipocytes. All three conditions (DNP, rotenone, and 3% oxygen) elevated hexose uptake by approximately twofold in 4 h relative to control cells. All three insults decreased cellular ATP levels rapidly. A subsequent recovery was observed within 1-2 h in the presence of DNP or 3% oxygen, probably as a result of anaerobic production of ATP through increased glucose uptake and glycolysis. DNP and rotenone elevated the content of GLUT-1 protein in isolated plasma membranes and decreased it in intracellular light microsomes, suggestive of translocation of this transporter isoform. No change in GLUT-4 protein distribution was detected. In contrast, 3% oxygen caused a marked specific increase in GLUT-1 protein in both plasma membranes and microsomes. Consistently, cycloheximide had no effect on the hexose transport responses to DNP or rotenone, but prevented the response to hypoxia. However, GLUT-1 mRNA and the total cell content of GLUT-1 protein were elevated by all three treatments. It is proposed that within the time frame studied, reductions in the energy charge may activate the glucose transport system in L6 myotubes and 3T3-L1 adipocytes by GLUT-1 protein biosynthesis and translocation. When both responses exist, the biosynthetic pathway is dispensable, and posttranslational mechanisms, including transporter translocation suffice to sustain the adaptive elevation in glucose transport activity for several hours.

2,4-Dinitrophenol↗

Calcium-antagonistic effects of carbamazepine in epilepsies and affective psychoses.

Carbamazepine (CBZ) is known to have beneficial effects in the treatment of epilepsies and in the prophylaxis of affective disorders. Since increased transmembrane calcium fluxes and intracellular calcium concentrations play a key role in the generation of epilepsies and possibly also in the development of these psychiatric disorders the effects of CBZ on epileptic discharges (elicited by caffeine, penicillin and low Mg2+) in CA3 neurons of hippocampal slices were compared with those of the organic calcium antagonist verapamil and found to be almost the same.

Animals↗

A passive transfer model of the organ-specific autoimmune disease, bullous pemphigoid, using antibodies generated against the hemidesmosomal antigen, BP180.

Subepidermal blistering associated with the human skin diseases bullous pemphigoid and herpes gestationis has been thought to be an IgG autoantibody-mediated process; however, previous attempts to demonstrate the pathogenicity of patient autoantibodies have been unsuccessful. An immunodominant and potentially pathogenic epitope associated with these blistering diseases has recently been mapped to the extracellular domain of a human epidermal antigen, BP180. Patient autoantibodies that react with this well-defined antigenic site failed to crossreact with the murine form of this autoantigen and thus could not be assayed for pathogenicity in a conventional passive transfer mouse model. As an alternative, rabbit polyclonal antibodies were generated against a segment of the murine BP180 protein homologous with the human BP180 autoantibody-reactive site and were passively transferred into neonatal BALB/c mice. The injected animals developed a subepidermal blistering disease that closely mimicked bullous pemphigoid and herpes gestationis at the clinical, histological, and immunological levels. Autoantibodies that recognize the human BP180 ectodomain are therefore likely to play an initiatory role in the pathogenesis of bullous pemphigoid and herpes gestationis.

Amino Acid Sequence↗

Experimental tracheal tube created with vascularized fascia.

An experimental fascial transferable bed was developed in the rabbit model. This tissue is reliable in bringing a viable mucosal graft inside the larynx. A vascular connective tissue sheet with full-thickness mucosa and autogenous cartilage for external support are needed. In this study a tracheal tube was preformed to study the use of autogenous cartilage as support for a circumferential lumen.

Animals↗

The role of glucose-6-phosphatase in the action of insulin on hepatic glucose production in the rat.

It has been suggested that regulation of glucose-6-phosphatase by insulin plays a role in the suppression of hepatic glucose production during feeding. We used hepatic glucose production (measured with the D-[3-3H]glucose infusion method) as an indicator of substrate flux through glucose-6-phosphatase in vivo. Compared with saline controls, insulin (7 mU.min-1 x kg-1, euglycemic clamp) suppressed hepatic glucose production virtually completely in both fasted (32.4 +/- 2.4 vs. -6.1 +/- 14 mumol.min-1 x kg-1) and fed (64.6 +/- 6.4 vs. 5.5 +/- 5.2 mumol.min-1 x kg-1) rats. Whereas hepatic glucose production was totally suppressed, [glucose-6-phosphate] in liver cytosol declined by only 27 and 35% in fasted and fed rats, respectively. Addition of hyperglycemia (10 mM) to the insulin infusion likewise fully suppressed hepatic glucose production (26.9 +/- 1.4 vs. -9 +/- 10 mumol.min-1 x kg-1 and 80.8 +/- 10.1 vs. -3.6 +/- 12.6 mumol.min-1 x kg-1 in fasted and fed rats, respectively), but [glucose-6-phosphate] again declined only modestly (21 and 27% in fasted and fed rats, respectively). This disproportionate suppression of hepatic glucose production could not be explained by cooperative substrate effects inasmuch as microsomal glucose-6-phosphatase isolated from saline- and insulin-treated rats followed Michaelis-Menten kinetics (Hill coefficient approximated 1). Acute insulin treatment of fasted rats in vivo did not reproducibly inhibit glucose-6-phosphatase activity assayed subsequently in isolated microsomes incubated in the absence of insulin.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Differential regulation of GLUT1 and GLUT4 glucose transporters in skeletal muscle of a new model of type II diabetes. The obese SHR/N-cp rat.

The obese diabetic SHR/N-cp rat is a newly developed strain that inherits obesity as an autosomal recessive trait. These rats display early-onset hyperinsulinemia and hyperglycemia, which are hallmarks of type II diabetes. This study was undertaken to determine the expression and the subcellular distribution of the GLUT1 and GLUT4 glucose transporters in skeletal muscle of obese diabetic SHR rats. D-glucose-protectable cytochalasin-B binding to subcellular membrane fractions of hindlimb muscles was used to determine glucose transporter number. GLUT1 and GLUT4 glucose transporter isotypes were detected using antibodies to the COOH-terminal region of the GLUT1 and GLUT4 proteins. Glucose transporter number was significantly lower (-40%) in crude unfractionated membranes of obese diabetic SHR than of lean SHR muscles. When crude membranes were fractionated to separate plasma membranes and the intracellular membranes containing glucose transporters, the number of cytochalasin-B binding sites was found to be markedly lower (-50%) in intracellular membranes and slightly but not significantly reduced (-20%) in plasma membranes of muscle from obese diabetic SHR compared with lean SHR rats. Western blot analysis revealed that a lower GLUT4 protein abundance (-40%) accounts for the reduced glucose transporter number in intracellular membranes of obese diabetic SHR compared with lean SHR muscles. GLUT4 protein content was also reduced by 50% in plasma membranes from obese SHR muscles relative to lean rat muscles.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

In vivo nuclear magnetic resonance spectroscopy of chicken embryos from two broiler strains of varying fat content.

In vivo nuclear magnetic resonance (NMR) imaging and spectroscopy techniques were used to monitor changes in P- and H-containing molecules in embryos of two broiler strains (30 and 31) differing genetically in fat content and ranging in age from 0 to 20 days of incubation. Chemical analysis showed that Strain 30 has more carcass fat than Strain 31 at market age (7 wk). Proton (1H) and 31P spectra were obtained on four eggs per strain at Days 0, 2, 4, 6, 8, 11, 12, 14, 16, 17, 19, and 20 of incubation. Fat:water, phosphomonoester (PME):phosphodiester (PDE), and adenosine triphosphate (ATP):PDE ratios were calculated. Chicks were hatched, grown to market weight (2,000 g for females and 2,300 g for males at 7 wk), and the whole intact carcasses were analyzed for crude fat. Hydrogen-1 NMR spectroscopy studies of incubated eggs indicated no significant difference (P > .05) in the fat:water ratio between the two strains. The difference in the PME:PDE ratios between the two strains as obtained by 31P-NMR spectroscopy over all days of incubation analyzed was not significant (P > .05); however, there was a significant difference in this ratio between the two strains at Days 4, 6, and 11. Up to Day 16, Strain 30 had a slightly, but not significantly (P > .05), higher ATP:PDE ratio as shown on 31P-NMR spectra, whereas after Day 17 the ATP:PDE ratio was significantly higher (P < .01) for Strain 31. Strain 31 birds had a significantly lower (P < .05) crude fat content. There was a significant difference (P < .05) in 7-wk carcass fat content between sexes, males having significantly (P < .01) less fat than females, which was correlated with a significantly higher (P < .01) ATP:PDE ratio in male embryos. It might be possible to use ATP:PDE ratios obtained during embryonic development by 31P-NMR to select strains of birds for low fat content at market weight and to distinguish between sexes during late embryonic development.

Adipose Tissue↗

[Purification and characterization of lipoteichoic acid from Streptococcus Sobronus]

Lipoteichoic acid (LTA) is amphipathic molecule found in cell surface of gram-positive bacteria.Although there were some hypothese about adhesion of LTA,the role of LTA in bacterial pathogenicety is still unclear.For this reason,a seria studies of LTA were carried out.In this study LTA was extracted by hot aqueous phenol from Strptococcus Sobronus 6715 grown in tryptic soy both supplemented with 5% sucrose for 17 hours,and was purfied on gel filtration by Sepharose 6B,chloroform-methanol precipitation and chromtography on DEAE-Sephacel.The purified LTA was characterized by FT-IR and 1H-NMR with standard sample of LTA (Sigma Co) as control.The results showed that LTA contains alanine ester and glycosyl residues which take important role in the amphipathic molecule.

Journal Article↗

Elevated levels of CD38+ CD8+ T cells in HIV infection add to the prognostic value of low CD4+ T cell levels: results of 6 years of follow-up. The Los Angeles Center, Multicenter AIDS Cohort Study.

A cohort of 98 HIV-infected initially AIDS-free homosexual men from the Multicenter AIDS Cohort Study (MACS) was followed for 6 years to investigate whether CD8+ cell subsets have prognostic value for progression to AIDS. In the present study, four subsets of CD8+ T cells that previously have been shown to be selectively elevated in HIV-infected asymptomatic persons, specifically the CD8+ T cell subsets that were CD38+, HLA-DR+, CD57+ and L-selectin negative (Leu8-), were measured. Forty-nine of the 98 developed AIDS. Prognostic value of these CD8+ cell subsets was evaluated using the proportional hazards model. Levels of both CD38+ CD8+ and Leu8- CD8+ cells individually had prognostic value for progression to AIDS. In contrast, CD57+ CD8+ and HLA-DR+ CD8+ cell subsets levels did not have prognostic value. After adjustment for level of CD4+ T cells, however, only the elevation in the CD38+ CD8+ cell subset had additional prognostic value. These results suggest that the level of CD38+ CD8+ cells could be used together with the CD4+ T cell level to more accurately predict progression to AIDS among HIV-infected men. These results provide further support for the observation that dramatic and progressive activation of CD8+ T cells in HIV infection occurs. The power of elevated levels of the CD38+ CD8+ subset to predict poor prognosis in this cohort suggests these CD8+ T cells reflect an immune stimulation that is ultimately unable to control disease progression.

ADP-ribosyl Cyclase↗

Hemodynamic effects of terazosin in congestive heart failure.

To determine the hemodynamic effects of a new alpha-1 blocker, terazosin, in congestive heart failure, 10 patients with this condition underwent hemodynamic testing at rest. Thirty minutes after the oral administration of 4 mg terazosin, mean right atrial pressure, mean pulmonary artery pressure and pulmonary capillary wedge pressure were found to be decreased, while the cardiac index was increased significantly. Improvement in hemodynamics persisted for more than 15 h, without an increase in heart rate. It is suggested that terazosin may have effective applications in the treatment of congestive heart failure.

Adolescent↗

[Effects of the stimulating point and non-point on human cerebral evoked potential and spinal cord evoked potential].

The experiments were carried out on 36 normal subjects. Pain stimulation points are at Neiguan or Hegu and pain stimulation nonpoints are nearby Neiguan or Hegu. The Record locations are at cortex and cervical vertebrae. The feature of evoked cerebral potential (ECP) and spinal cord evoked potential (ESP) by pain stimulation the points or non-points were studied. The results have been obtained. 1. Pain stimulation point ECP generally consists of wave P1, N1, P2, N2, P3, N3, P4 and N4. There is a certain amplitude and latency of each wave. 2. The elements of pain stimulation non-point ECP are Similar to point ECP in the feature. Comparing Pain stimulation Point ECP with non-point ECP, the amplitude of wave P3-N3 of stimulation point ECP increases (P < 0.05). 3. Pain stimulation Point ESP and non-point ESP consist of wave P1, N1 and P2, N3. Comparing Pain stimulation point with non-point there are no significant changes in the amplitude and the latency of ESP.

Acupuncture Points↗