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Biomedical subjects

Q Li

Publications and source records attributed to Q Li.

At least 505 records · Page 28Linked to original sources
PubMed1998

Inflammatory response in the early stages of wound healing after excimer laser keratectomy.

OBJECTIVE: To evaluate the inflammatory response and its potential role in the early stages of corneal wound healing after excimer laser keratectomy. MATERIALS AND METHODS: Lewis rats underwent excimer keratectomy using a 193-nm excimer laser. The central corneas were ablated in 3 depths: group A, epithelium; group B, superficial stroma; or group C, deep stroma. Eyes were harvested 1, 12, 24, and 36 hours, and 1 week after the rats were killed. Immunohistochemistry was used to test frozen sections with monoclonal antibodies of various inflammatory cellular markers. RESULTS: Reepithelialization was observed at 12 hours in group A, and at 24 hours in groups B and C. Regenerated epithelium covered the denuded corneal surface in groups B and C after 1 week. The expression of major histocompatibility complex II antigen was detected in infiltrating cells, corneal epithelial cells, and endothelial cells 1 hour after surgery. Only a few macrophages and Langerhans cells were in the limbus at baseline. Macrophages migrated from the limbus to the corneal ablation zone and increased 2-fold after 36 hours in all 3 groups compared with baseline. Occasional lymphocytic infiltration was identified after 25 to 36 hours. CONCLUSION: Macrophages play an active role in the wound healing after laser keratectomy and may contribute to transient corneal haze.

Animals↗
PubMed1998

Overexpression of the 77-kD echinoderm microtubule-associated protein (EMAP), a WD-40 repeat protein, in baculovirus-infected Sf9 cells.

The purpose of this study was to test whether any assembly-promoting microtubule-associated protein (MAP) would bundle microtubules and induce process formation in recombinant baculovirus-infected Sf9 cells, in particular, whether a non-neural MAP from a normally rounded cell would produce cellular asymmetries. To carry out these experiments, we constructed a recombinant baculovirus that expressed the full-length 77-kD EMAP, an abundant MAP that localizes to the mitotic spindle of cleavage-stage sea urchin embryos and to the interphase array of microtubules in adult coelomocytes. Expression of EMAP in Sf9 cells had no detectable effect on cellular morphology, microtubule organization, or stability. These results indicate that process formation in Sf9 cells is MAP specific.

Animals↗
PubMed1998

Development of a condensed locus control region cassette and testing in retrovirus vectors for A gamma-globin.

Retrovirus vectors for A gamma-globin are being developed for the treatment of beta chain hemoglobinopathies. Toward the goal of achieving therapeutic expression levels, core elements of the beta-globin locus control region (LCR) hypersensitive sites (HS) were screened for enhancer activity in erythroid MEL and K562 cell lines using a drug-resistant colony assay. When used alone, core elements of HS1, HS3, and HS4 showed no activity and a fragment for HS2 showed only modest activity in the colony assay. However, a 1.1 kb combination of fragments for HS2, HS3, and HS4 (termed a nLCR) enhanced colony formation 17-fold in K562 cells and 94-fold in MEL cells. Addition of an HS1 fragment enhanced nLCR activity only modestly in MEL cells. When linked to a beta-globin gene, the 1.1 kb nLCR enhanced globin mRNA expression to 82% per copy of mouse alpha-globin in transfected MEL cells. Inclusion of a nLCR in retrovirus vectors containing a beta-globin promoter and various A gamma-globin gene expression cassettes resulted in extreme genetic instability and reduced titers. Specific deletions were abrogated by removing homologous sequences, but random recombinations were still observed at significant frequencies. In MEL cells containing intact provirus, A gamma-globin mRNA produced by an optimal vector containing the nLCR was only 2-fold higher (8.5% vs. 3.9% per copy of mouse alpha-globin) compared to the same vector without the nLCR. These data suggest that vector elements detract from the ability of the nLCR to enhance expression of the beta pr.A gamma cassettes.

Animals↗
PubMed1998

Modulation of interferon gamma induced increases in cathepsin B in THP-1 cells by adrenergic agonists and antagonists.

In order to investigate the possible modulation of macrophage function by the autonomic nervous system, the effect of adrenergic agonists and antagonists on interferon (IFN)-gamma-induced increases in cathepsin B (CB) in a macrophage-like cell line was studied. It has been shown previously that IFN-gamma induces increased CB activity in phorbol myristate acetate (PMA)-primed THP-1 cells. Isoproterenol (ISO) (10 micrometers), a mixed beta-receptor agonist, increased the induction of CB activity in the cells but norepinephrine (10 micrometers) and epinephrine (10 micrometers), the alpha and beta receptor agonists, had little effect. The addition of the mixed alpha-receptor antagonist phentolamine (10 micrometers) had no effect on ISO induced increases but the mixed beta-receptor antagonist propranolol (10 micrometers) and the selective beta1-receptor antagonist atenolol produced significant inhibition. These results suggest that the activation of beta-receptors could be involved in the induction of CB activity in macrophages and provide a possible mechanism for the regulation of macrophage effector function by the autonomic nervous system. Dibutyryl cAMP (1 mm) alone also induced increases in CB in THP-1 cells, and H-89 or HA1004 abrogated the effect of dibutyryl cAMP, suggesting that the effect of ISO on CB could be through the elevation of cAMP and the activation of cAMP-dependent protein kinases.

Adrenergic Agonists↗
PubMed1998

Intraarterial urokinase produces significant attenuation of infarction volume in an embolic focal ischemia model.

A number of models of focal ischemia have been created to mimic acute middle cerebral artery (MCA) occlusion. In the present series of experiments, we report our observations on the thrombin model of MCA occlusion and the neuroprotective effects of intraarterial thrombolysis with two doses of urokinase (2500 and 5000 units/kg). In all experiments male Wistar rats were used and the animals were allowed to recover for 48 h before assessment of neurobehavioral performance on a four-point scale. The extent of cerebral hemispheric damage was calculated as the percentage of brain infarction using TTC staining. Occlusion of the MCA was effected by the introduction of an autologous blood clot into the internal carotid artery (ICA) approximately 2 mm from the origin of the MCA. This clot was formed by the drawing of 10 microl of blood into a bovine thrombin (20 microg per animal) containing intraarterial catheter, which was inserted into the right ECA. After standing for 15 min to allow clot formation, the catheter was advanced gently through the ICA to the site of injection. MCA occlusion produced a consistent large infarction in all animals. Urokinase infusion (i.a. ) was started 2 h after arterial occlusion in the initial series. In animals treated with low dose urokinase infusion there was mild protection. Animals treated with high dose urokinase infusion showed a highly significant improvement in the motor recovery and a decrease in the extent of infarction compared to control animals. In the final group, the infusion of urokinase was delayed for 3 h. While producing protection in some animals, it also produced intracerebral hemorrhage in two of eight animals. Thus delay of infusion to 180 min increased the risk of hemorrhage. This model may in the future be used to test the protective effects of combination therapy with thrombolysis and neuroprotective medications.

Animals↗
PubMed1998

Cholecystokinin induces Fos expression in the brain of the Japanese quail.

Systemic administration of cholecystokinin (CCK) inhibits feeding in birds. However, the signaling pathway through which CCK induces this effect is unknown, and its role as a natural satiety signal is controversial. To address these issues, we used immunocytochemistry for the immediate-early gene protein Fos to localize sites of neuronal activation in the brain of Japanese quail (Coturnix japonica) after CCK treatment. Food intake was inhibited in a dose-dependent manner following intraperitonal (i.p.) injection of CCK, with an effective dose range of 1-50 micrograms/kg. To test the hypothesis that CCK induces a distinct pattern of Fos-like immunoreactivity (FLI) in the brain, we compared FLI in birds given CCK (20 micrograms/kg, i.p.) with that in birds given a nonspecific chemical inhibitor of feeding, lithium chloride (LiCl, 40 mg/kg, i.p.), at doses that reduce feeding to a similar level (30% of saline controls). FLI-positive cell nuclei were counted in 14 brain regions after administration of CCK, LiCl, or saline. CCK uniquely induced FLI in the paraventricular, infundibular, periventricular hypothalamic, and medial mamillary nuclei of the hypothalamus. However, CCK and LiCl both induced a comparable pattern of FLI in the hindbrain, with strong staining in the nucleus of the solitary tract and dorsal motor nucleus of the vagus. These findings demonstrate the ability of CCK to activate the central nervous system in birds and suggest that the peptide exerts specific actions in the hypothalamus. However, the possibility that the FLI observed may have arisen through nonspecific effects of CCK on gastrointestinal physiology cannot be discounted.

Animals↗
PubMed1998

Recombinant adenovirus encoding gp100 modulates experimental melanin-protein induced uveitis (EMIU).

Experimental melanin-protein induced uveitis (EMIU) is a T-cell mediated autoimmune uveitis induced by immunization with bovine uveal melanin protein. Gp100, a melanocyte lineage-specific protein, is identified as a human melanoma antigen. A recombinant adenovirus construct encoding gp100 (Ad2CMV-gp100) has been used as a vaccine for cancer therapy. This study examines the effect of Ad2CMV-gp100 on EMIU. To induce EMIU, rats were injected intraperitoneally on day 7 before immunization with ad2CMV-gp100, control adenovirus encoding LacZ (Ad2CMV-LacZ), or no virus. On day 21 after immunization, the right eye was processed for histology and the left eye was analysed for cytokines by quantitative reverse transcriptase-polymerase chain reaction. Western blot analysis showed that uveal melanin-protein contains gp100. In three independent experiments, ocular inflammation was significantly suppressed, and expression of ocular IL-12p40 mRNA was much lower in the rats which received Ad2CMV-gp100 before immunization than in those that received Ad2CMV-LacZ or no virus. No abnormalities developed in rats which received Ad2CMV-gp100 or Ad2CMV-LacZ alone. Therefore, Ad2CMV-gp100 injection prevents the development of EMIU, at least in part, through cytokine regulation.

Adenoviridae↗
PubMed1998

Reversal of hydroquinone-mediated suppression of T cell proliferation by transfection of the M2 subunit of ribonucleotide reductase.

Hydroquinone (HQ) is a benzene derivative that is found in large quantities in cigarette tar as a result of the pyrolysis of tobacco flavinoids. HQ is a potent inhibitor of T cell proliferation, causing an immediate and complete cessation of DNA synthesis in IL-2-dependent human T lymphoblasts and Jurkat T cells without loss of cell viability. Previous studies from our laboratory demonstrated that the antiproliferative effects of RQ could be partially reversed by the addition of deoxyribonucleosides, but not by the corresponding ribonucleosides, suggesting that HQ might inhibit ribonucleotide reductase. In the present study, the molecular mechanism behind this observation was further investigated. Jurkat T cells were stably transfected with a pMEP4 expression vector containing the gene for the M2 subunit of ribonucleotide reductase under transcriptional control of the human metallothionein IIA promoter. M2-transfected Jurkat T cells exhibited a greater than three-fold increase in resistance to HQ compared to untransfected cells or cells transfected with the M2 gene in the reverse orientation. HQ resistance was associated with an increased level of M2 protein detected by Western blot. These results suggest that the benzene derivative inhibits lymphocyte proliferation by inhibiting ribonucleotide reductase.

Blotting, Western↗
PubMed1998

Pharmacokinetics and biodistribution of radioimmunoconjugates of anti-CD19 antibody and single-chain Fv for treatment of human B-cell malignancy.

The comparative advantages and disadvantages of intact antibodies and single-chain Fv as immunotoxins and radioimmunoconjugates have been widely discussed but not directly compared. In this study, the in vivo properties of anti-CD19 B43 monoclonal antibody and its derived single-chain Fv (FVS191) were studied in athymic nude mice bearing CD19-positive human lymphomas. B43 mab and FVS191 were labeled with iodine-125 using iodine-beads, and immunoreactivities were determined to be 57% and 72%, respectively. Scatchard analysis showed a similar high affinity for both. The results of pharmacokinetic studies revealed that FVS191 had a rapid biphasic clearance from the circulation (T1/2alpha=2.5 min, T1/2beta=3.7 h); The T1/2alpha and T1/2beta phases of B43 mab were determined to be 0.72 h and 57 h respectively. Biodistribution studies compared the uptake of labeled antibodies by CD19-positive and by CD19-negative tumors. The peak percentages of injected dose were 5.7% at 12 h for B43 and 2.45% at 1 h for FVS191. Radiolocalization indices (RI) demonstrated tumor-specific uptake for both, but higher uptake for B43. The optimal RI was seen at 15 min for FVS191 and 6 h for B43. FVS191 was unstable in vivo, approximately 50% of the injected dose being degraded in blood in 100 min. Radioactivity detected in the urine was present mainly as the deiodinized form of FVS191. The results suggest that B43 mab is favored over FVS191 in biodistribution properties and in vivo stability. Because B43 Mab showed early tumor-specific uptake, high RI values, and favorable tissue-to-blood ratios, it is a potential candidate for radioimmunotherapy and immunotoxin therapy of B-cell leukemia and lymphoma.

Animals↗
PubMed1998

Cellular localization of neuropeptide Y mRNA and peptide in the brain of the Japanese quail and domestic chicken.

Neuropeptide Y (NPY) has been implicated in the control of a number of physiological functions in birds including food intake and reproduction. In the present study, sites of NPY synthesis were localized in the brains of Japanese quail and domestic chickens by in situ hybridization histochemistry using a digoxigenin-labelled riboprobe. NPY mRNA was detected in three main cell groups in both species. The most prominent group was associated with structures in the lateral thalamus including the anterior lateral thalamic nucleus, lateral forebrain bundle, rotund nucleus, pretectal nucleus and occipitomesencephalic tract. Other major cell groups were detected in the hippocampus, and in the caudal linear nucleus and raphe nucleus of the brainstem. NPY mRNA was also present in the piriform cortex and taenial nucleus. Double-labelling of NPY mRNA and peptide was demonstrated in individual cells of the hippocampal, thalamic and brainstem cell groups, suggesting that NPY is synthesized and stored in these areas. However, the identity of other cell groups, notably in the hyperstriatal, archistriatal and neostriatal regions of the telencephalon, which exhibit NPY-immunoreactive cell bodies but no NPY mRNA, remains to be determined.

Animals↗
PubMed1998

A comparison of the oxytocin and vasopressin responses to the 5-HT1A agonist and potential anxiolytic drug alnespirone (S-20499).

The effect of the serotonin1A (5-HT1A) agonist alnespirone (S-20499) on the secretion of both oxytocin and vasopressin was examined in the same conscious, unrestrained male rats. The dose-response and time-course effects on the secretion of oxytocin and vasopressin revealed that alnespirone stimulated oxytocin in a dose-dependent manner, but did not increase vasopressin secretion. Time of maximal effect following injection of alnespirone (5 mg/kg, i.p.) was as early as 15 min postinjection, with significant stimulation persisting for 30 min. Pretreatment with a low dose of the 5-HT1A/beta-adrenoceptor antagonist (-)-pindolol (0.3 mg/kg, s.c.), 30 min prior to injection of alnespirone (0, 2, 5, and 10 mg/kg, i.p.) shifted the dose-response curve to the right and inhibited the effect of alnespirone on plasma oxytocin concentration. Furthermore, pretreatment with a low or a high dose of the 5-HT1A/2A/dopamine D2 antagonist spiperone (0.01 or 3 mg/kg, s.c.) dose dependently shifted the alnespirone dose-response curve effect of alnespirone to the right. None of these drugs, alone or in combination, altered plasma vasopressin levels. These studies suggest that 5-HT1A receptor mechanisms mediate the effect of alnespirone on the secretion of oxytocin. Furthermore, these studies suggest that 5-HT1A receptor mechanisms do not participate in the serotonergic regulation of vasopressin secretion.

Adrenergic beta-Antagonists↗
PubMed1998

Dose-dependent brainstem neuropathology following repeated arteether administration in rats.

Histopathological effects of the artemisinin antimalarial, beta-arteether, were evaluated in rats. Arteether (3.125-12.5 mg/kg/day, IM, in sesame oil) was administered for 7 consecutive days. Seven days following the last injection, histological evaluation of the brainstem was performed. Rats treated with 12.5 mg/kg showed significant neuropathology, including chromatolysis, in the nucleus trapezoideus and nucleus superior olive. To a lesser extent, neuropathology was present in the nucleus ruber. Mild neuropathology was also detected in other brainstem regions examined. Although no statistically significant neuropathology was found for the groups treated with 6.25 mg/kg/day and 3.125 mg/kg/day, substantial neuropathology was observed in a single rat in each of these treatment conditions. These results confirm and extend previous studies demonstrating brainstem neurotoxicity from artemisinin antimalarials. Furthermore, these results suggest that, in rats, brainstem auditory pathways may be particularly vulnerable. Early detection of arteether neuropathology may, therefore, require examination of auditory functions.

Animals↗
PubMed1998

Identification and determination of phase I metabolites of propafenone in rat liver perfusate.

Propafenone (PF) is a class 1C antiarrhythmic agent. To study the mechanisms of PF interactions with dietary nutrients in isolated, perfused rat livers, metabolites of PF in liver perfusate were identified and an analytical method was developed for these metabolites plus parent drug. Identification of phase I metabolites was performed using HPLC/MS equipped with a Lichrospher RP-18 column and tandem mass spectrometry (MS/MS) with electrospray and atmospheric pressure chemical ionizations. Three major metabolite peaks, whose protonated molecular ions were m/z 358, 358 and 300, were identified as a propafenone derivative hydroxylated in the omega-phenyl ring (omega-OH-PF), 5-hydroxypropafenone (5-OH-PF), and N-despropylpropafenone (N-des-PF). The levels of omega-OH-PF, 5-OH-PF, N-des-PF and PF were determined simultaneously by HPLC with UV detection at 210 nm and a mobile phase of 0.03% triethylamine and 0.05% phosphoric acid in water-acetonitrile-methanol (45:20:35, v/v/v) after extraction with 5 ml diethyl ether at pH 10.0 and evaporation of solvent under nitrogen. The results revealed that omega-OH-PF, which was not found in humans, was the major metabolite of PF in rat liver perfusate, not 5-OH-PF which is the major metabolite in human plasma.

Animals↗
PubMed1998

Membrane compartmentation is required for efficient T cell activation.

The plasma membrane of mammalian cells contains detergent-resistant membrane rafts enriched in glycosphingolipids and cholesterol. Although several important signaling molecules have been found in such rafts, evidence documenting a functional role for their localization has been scarce. Using a fractionation scheme that preserves tyrosine phosphorylation, we show that T cell activation leads to a striking compartmentation in the rafts of activated T cell receptor and associated signal-transducing molecules. Conditions that reversibly disrupt raft structure either by dispersing their contents or by forcing their internalization reversibly disrupt the earliest steps of T cell activation. Thus, raft integrity is a prerequisite for efficient T cell receptor signal transduction.

Animals↗
PubMed1998

Palmitoyl derivatives of interferon alpha: potential for cutaneous delivery.

Palmitoyl derivatives of interferon alpha2b (p-IFNalpha) were prepared by covalent attachment of the fatty acid to lysine residues in the protein through a reaction with N-hydroxysuccinimide palmitate ester. The p-IFNalpha was characterized by capillary electrophoresis (CE), mass spectrometry (MS), SDS-PAGE, and antiviral assay. Flow-through diffusion cells and human breast skins were used to measure cutaneous and percutaneous absorption. Formation of p-IFNalpha derivatives was demonstrated by CE to be dependent on reaction time and reagent: protein ratio. Electrospray MS of the crude p-IFNalpha mixture indicated three populations of IFNalpha derivatives with 10, 11, and 12 palmitoyl substitutions. The addition of palmitoyl residues to IFNalpha under the conditions described reduced the antiviral specific activity by 50%. However, the cutaneous absorption of p-IFNalpha was about 5-6 times greater than the parent protein. The amount of p-IFNalpha and IFN alpha in whole skin after 24 h of treatment was 2.106 +/- 1.216 microg/cm2 and 0.407 +/- 0.108 microg/cm2, respectively. Approximately two times higher flux was detected for p-IFNalpha compared to the nonfatty acylated IFNalpha. The total amount of drug diffused in 24 h was also approximately two times higher for the p-IFNalpha. The results indicate a potential for using fatty acylated derivatives of IFN alpha for dermal and transdermal delivery.

Acylation↗
PubMed1998

Z/I and A-band lattice spacings in frog skeletal muscle: effects of contraction and osmolarity.

A-band and Z-line/I-band lattice spacings were measured by small-angle X-ray diffraction from relaxed and isometrically-contracting whole frog sartorius muscles with lattice spacings reduced or swollen by changing the osmolarity of the bathing solution. A-band spacing increased by approximately 3% upon isometric contraction at reduced lattice spacings (245-356 mOsm) and decreased by approximately 1% at swollen spacings (172 mOsm), similarly to the behaviour of skinned muscles upon changing from the relaxed state to rigor. The Z/I lattice underwent a significant lattice expansion (3-8%) upon isometric contraction at all osmolarities, in qualitative agreement (but quantitative disagreement) with results from electron microscopy on mammalian skeletal muscle. Lattice areas calculated for the Z/I and A-band lattices indicate a barrel-shaped sarcomere in the resting state, which may provide a partial explanation for how longitudinal forces produced in the A-band can produce a radial expansive force in the Z-line during contraction. The radial component of cross-bridge stiffness was calculated from the A-band data for contracting muscle, using a lattice stability model incorporating structural, osmotic and electrostatic forces. The calculations gave a radial cross-bridge stiffness during contraction of about 9 x 10(5) N m-2, and outward radial force per thick filament in normal Ringer's solution of 6 x 10(-9) N, corresponding to a radial force per cross-bridge of 10(-11) N.

Animals↗
PubMed1998

Mechanism of block by tedisamil of transient outward current in human ventricular subepicardial myocytes.

1. Tedisamil is a new antiarrhythmic drug with predominant class III action. The aim of the present study was to investigate the blocking pattern of the compound on the transient outward current (I(to)) in human subepicardial myocytes isolated from explanted left ventricles. Using the single electrode whole cell voltage clamp technique, I(to) was analysed after appropriate voltage inactivation of sodium current and block of calcium current. 2. Tedisamil reduced the amplitude of peak I(to), but did not affect the amplitude of non-inactivating outward current. The drug accelerated the apparent rate of I(to) inactivation. The reduction in time constant of I(to) inactivation depended on drug concentration, the apparent IC50 value was 4.4 microM. 3. Tedisamil affected I(to) amplitude in a use-dependent manner. After 2 min at -80 mV, maximum block of I(to) was reached after 4-5 clamp steps either at the frequency of 0.2 or 2 Hz, indicating that the block was not frequency-dependent in an experimentally relevant range. Recovery from block was very slow and proceeded with a time constant of 12.1+/-1.8 s. Also in the presence of drug, a fraction of channels recovered from inactivation with a similar time constant as in control myocytes (i.e. 81+/-40 ms and 51+/-8 ms, respectively, n.s.). 4. From the onset of fractional block of I(to) by tedisamil during the initial 60 ms of a clamp step, we calculated k1 = 9 x 10(6) mol(-1) s(-1) for the association rate constant, and k2 = 23 s(-1) for the dissociation rate constant. The resulting apparent KD was 2.6 microM and is similar to the IC50 value. 5. The effects of tedisamil on I(to) could be simulated by assuming a four state channel model where the drug binds to the channel in an open (activated) conformation. It is concluded that in human subepicardial myocytes tedisamil is an open channel blocker of I(to) and that this effect probably contributes to the antiarrhythmic potential of this drug.

Action Potentials↗