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Biomedical subjects

D Liu

Publications and source records attributed to D Liu.

At least 865 records · Page 48Linked to original sources
PubMed1977

Effect of procaine HCLl on ATP: calcium-dependent alterations in red cell shape and deformability.

Procaine hydrochloric acid, a cationic anesthetic, although unable to prevent the effect of calcium ionophore A23187 on erythrocytes, inhibited the discocyte--echinocyte transformation, increased viscosity, and decreased filterability of red cells undergoing ATP depletion. The effects were abolished by washing ATP-depleted, procaine HCl-treated red cells prior to these determinations. Procaine HCl had no effects on volume, incubated osmotic fragility, or monovalent cation composition of ATP-depleted red cells. The drug increased 45Ca uptake by ATP-depleted red cells but did not change the fraction of membrane-bound calcium. Sodium dodecyl sulfate acrylamide gel electrophoresis of membrane proteins from ATP-depleted red cells revealed formation of high molecular weight protein complexes, which were not formed when biconcave shape and ATP content were maintained by incubation with adenine (0.54 mM) and inosine (12.7 mM); Formation of these complexes was not prevented when the biconcave shape was maintained by procaine HCl. It was concluded that the maintenance of the biconcave shape and normal deformability during ATP depletion by procaine HCl was not related to a displacement of membrane-bound calcium and inhibition of ATP-dependent rearrangement of red cell membrane proteins.

Adenosine Triphosphate↗
PubMed1975

Primary structure of porcine pepsin. III. Amino acid sequence of a cyanogen bromide fragment, CB2A, and the complete structure of porcine pepsin.

The complete amino acid sequence of porcine pepsin (EC 3.4.4.1) was constructed from the sequence of five cyanogen bromide fragments. The sequence of one of these fragments, CB2A, is reported here. The sequences of 4 other fragments are known from previous work. Porcine pepsin contains 327 residues with three structural variants. The active center aspartyl residue, which reacts with 1,2-epoxy-3-(p-nitrophenoxy)propane (Chen, K. C. S., and Tang, J. (1972) J. Biol. Chem. 247, 2566-2574), is located at residue 32. Another active site aspartyl residue, which reacts with diazo inactivators (Bayliss, R. S., Knowles, J. B., and Wybrandt, G. B. (1969) Biochem. J. 113, 377-386, IS LOCATED AT RESIDUE 215. The sequences around these 2 aspartyl residues are apparently homologous to each other. The sequences around the tryptophanyl residues at positions 39, 141, 181, and 300 are also homologous to one another. These homologous sequences could be genetic in origin. Fragment CB2A which contains 119 residues was constructed from the peptide sequences resulting from six proteolytic digestions and chemical cleavage at tryptophanyl bonds.

Amino Acid Sequence↗
PubMed1975

Strike-through of moist contamination by woven and nonwoven surgical materials.

A test is described which correlates the stress of stretching surgical gown and drape material with moist bacterial strike-through. By application of this test to a number of woven and nonwoven surgical gown and drape materials, it was found that not all of these materials, either woven or nonwoven, are impermeable to moist contamination for equal periods of time. Nonwoven disposable materials now in use range from those which remain impermeable to moist bacterial permeation through all tests while some remain impermeable for limited periods of time, and others almost immediately permeable to moist bacterial penetration. The same situation holds for woven materials. Under conditions of our test, Quarpel treated Pima tight-woven cotton cloth was impermeable to moist bacterial strike-through, through up to 75 washing and sterilizing cyclings, while ordinary linen and untreated Pima cloth permitted bacterial permeation almost immediately. These results have significance in lengthy wet surgical operations.

Bacteria↗
PubMed1973

Amino-acid sequence of porcine pepsin.

As the culmination of several years of experiments, we propose a complete amino-acid sequence for porcine pepsin, an enzyme containing 327 amino-acid residues in a single polypeptide chain. In the sequence determination, the enzyme was treated with cyanogen bromide. Five resulting fragments were purified. The amino-acid sequence of four of the fragments accounted for 290 residues. Because the structure of a 37-residue carboxyl-terminal fragment was already known, it was not studied. The alignment of these fragments was determined from the sequence of methionyl-peptides we had previously reported. We also discovered the locations of activesite aspartyl residues, as well as the pairing of the three disulfide bridges. A minor component of commercial crystalline pepsin was found to contain two extra amino-acid residues, Ala-Leu-, at the amino-terminus of the molecule. This minor component was apparently derived from a different site of cleavage during the activation of porcine pepsinogen.

Amino Acid Sequence↗
PubMedDate not supplied

Characterization of local inflammatory response in an isolated lung perfusion model.

BACKGROUND: Current phase I trials of isolated lung perfusion for treatment of pulmonary metastases have an arbitrarily determined length of perfusion. Our objective was to examine the temporal course of the local and distant inflammatory response as a function of the length of perfusion (ischemia) and subsequent reperfusion in an equivalent animal model. METHODS: Sixty male Fischer 344 rats were randomized into four groups (n = 15). Each group underwent left isolated lung perfusion with buffered Hespan for 10, 30, 60, or 90 minutes. Subsequently, two subgroups of five animals within each group were allowed to reperfuse for 1 or 3 hours, respectively. Non-perfused right lung was used as control. At each time point, lung specimens were assayed for TNF-alpha by ELISA and histologic sections were examined. RESULTS: There was no significant difference between the left and right lung tissue levels of TNF-alpha at the termination of the ischemic period. However, on reperfusion, the left lung TNF-alpha levels increased significantly above the ischemia baseline in all groups, with a greater magnitude of rise in the groups with 60 and 90 minutes of preceding ischemia (12,757 +/- 1985 vs. 3524 +/- 494 pg/g, and 16,914 +/- 1657 vs. 6530 +/- 1104 pg/g, respectively; p < 0.05). There was no significant elevation in tissue levels of TNF-alpha in the right lung. Histologic changes consistent with early pulmonary edema were first detected at 12 hours following onset of reperfusion. CONCLUSIONS: Reperfusion following prolonged pulmonary ischemia during isolated lung perfusion results in a significant elevation of local tissue levels of TNF-alpha and may render the perfused lung vulnerable to the adverse effects of the inflammatory cascade.

Animals↗
PubMedDate not supplied

Blood flow dynamics in different layers of the somatosensory region of the cerebral cortex on the rat during mechanical stimulation of the vibrissae.

The present work reports studies of the quantitative spatial and temporal characteristics of changes in local blood flow in different layers of the somatosensory cortex of rats during adequate mechanical stimulation of the vibrissae. Studies were performed using 34 Wistar rats. Skull trepanning was performed under urethane (1 g/kg) anesthesia. Television-guided microscopy was used to introduce a set of three platinum electrodes (100 microns in diameter, with tip diameters of 30-40 microns) into the somatosensory cortex projection zone of the vibrissae. The first and third electrodes were positioned in cortical layers I-III and IV-VI and the central electrode was used to generate hydrogen within the tissue. Electrode positions were confirmed histologically after experiments. Animals were placed on artificial ventilation and one or all vibrissae were stimulated at a frequency of 3 Hz for 60 sec, with interstimulus intervals of 3 min. Changes in the local blood flow were measured during stimulation and for 1 min afterwards, using the hydrogen clearance method, and brain tissue impedance was also measured. There was a small (up to 5-7%) reduction in blood flow in the first seconds of stimulation, which was followed 15-25 sec later by an increase and subsequent return to initial when stimulation stopped. The increases in blood flow during stimulation of all vibrissae were by 24.2 +/- 6.7% (n = 36) in layers IV-VI and 24.5 +/- 5.6% (n = 34) in layers I-III; increases in response to stimulation of single vibrissae were by 19.4 +/- 7.4% (n = 28) and 17.8 +/- 6.4% (n = 28) respectively. The dynamics of impedance changes corresponded to those of blood flow changes. Thus, heterogeneity was found in changes of local brain blood flow in different layers of the somatosensory cortex during increases in cortical functional activity.

Animals↗
PubMedDate not supplied

An experimental model combining microdialysis with electrophysiology, histology, and neurochemistry for studying excitotoxicity in spinal cord injury. Effect of NMDA and kainate.

We used an experimental model that we previously developed to characterize the damage caused by the agonists of glutamate receptors, N-methyl-D-aspartate (NMDA) and kainate, in the spinal cord in vivo, thereby testing further the utility of this model. Microdialysis was used to administer the toxins and to sample the release of other substances in response to these agents. The blockage of electrical conduction was monitored by recording the amplitudes of evoked potentials during administration of the damaging substances, and damage was assessed by postmortem histological examination. The released amino acids in microdialysates were measured by HPLC. Administration of 5 mM NMDA + 5 mM kainate into the gray matter blocked most postsynaptic responses and caused the release of amino acids. Administration of 10 mM NMDA and 10 mM kainate significantly destroyed cell bodies near the fiber. The advantage of this model is that histological, neurochemical, and electrophysiological parameters were obtained in the same experiment.

Amino Acids↗
PubMedDate not supplied

Development of gene probes of Dichelobacter nodosus for differentiating strains causing virulent, intermediate or benign ovine footrot.

Seven Dichelobacter nodosus genomic DNA clones including six specific for virulent and one for benign strains were identified. A collection of 96 footrot isolates, which in turn comprised 27 virulent isolates showing elastase activity at 7 days, 25 high intermediate isolates with elastase activity at 14 days, 24 low intermediate isolates with elastase activity at 21-28 days and 20 benign isolates with no elastase activity at up to 28 days, were used to assess these clones. Of the six virulent specific clones, five (pV238-83, pV405-239, pV470-65, pV470-145 and pV470-178) reacted with 27 virulent isolates, and 12 of 25 high intermediate isolates, but none of 24 low intermediate isolates and 20 benign isolates in dot blot hybridization. The other virulent-specific clone (pV470-13) recognized all 27 virulent and 25 intermediate isolates, and 22 of 24 low intermediate isolates and three of 20 benign isolates in dot blot hybridization. By contrast, the benign-specific clone (pB645-335) detected all 20 benign isolates and 24 low intermediate isolates, and also 13 of 25 high intermediate isolates, but none of 27 virulent isolates in dot blot hybridization. Southern hybridization analysis indicated that whereas clones pV238-83, pV405-239 and pV470-178 bound a Sau3A band of 0.5 kb, clones pV470-65 and pV470-145 recognized two Sau3A bands of 0.7 and 0.5 kb in virulent strains of serogroups A to I. However, clone pV470-13 detected a Sau3A band pattern in virulent strains different from those recognized by the other five virulent specific-clones. Besides showing a distinct Sau3A band pattern in intermediate strains, pV470-13 also reacted with three benign strains that showed binding with it in dot blot hybridization. The benign-specific clone pV645-335 detected a Sau3A band of 0.5 kb in both intermediate and benign strains of serogroups A to I. Thus the combination of pV470-13 and pB645-335, or any other virulent-specific clone, would clearly differentiate among D. nodosus strains causing virulent, intermediate or benign footrot.

Animals↗
PubMedDate not supplied

Analysis of the ligand binding site of the 5-HT3 receptor using site directed mutagenesis: importance of glutamate 106.

The 5-HT3 receptor is a ligand-gated ion channel with significant structural similarity to the nicotinic acetylcholine receptor. Several regions that form the ligand binding site in the nicotinic acetylcholine receptor are partially conserved in the 5-HT3 receptor, presumably reflecting the conserved signal transduction mechanism. Specific amino acid differences in these regions may account for their distinct ligand recognition properties. Using site-directed mutagenesis, we have replaced one of these residues, glutamate 106 (E106), with aspartate (D), asparagine (N), alanine (A) or glutamine (Q) and characterized the ligand-binding and electrophysiological properties of the mutant receptors after transient expression in HEK-293 cells. The affinity for the selective 5-HT3 receptor antagonist [3H]GR65630 was decreased 14-fold in the mutant E106D (Kd = 3.69 +/- 0.32 nM) when compared to wildtype (WT, E106) 5-HT3 receptor (0.27 +/- 0.03 nM), while the affinity for E106N was unchanged (0.42 +/- 0.07 nM, means +/- SEM, n = 3-10). Decreased affinities for both E106D and E106N were observed for the antagonists granisetron, ondansetron and renzapride and for the agonists 5-HT (130- and 30-fold) and 2-methyl-5-HT (250- and 20-fold), respectively. Both mutants still formed 5-HT-activatable ion channels, but the high Hill coefficient of the concentration effect curves in wildtype (2.0) was decreased to unity in both cases. The EC50 of 5-HT was increased seven-fold in E106N (8.7 microM) when compared to wildtype (1.2 microM), but unchanged in E106D, and the potency of the antagonist ondansetron for both mutants was decreased. E106A and E106Q expressed poorly preventing a detailed characterization. These data suggest that E106 contributes to the ligand-binding site of the 5-HT3 receptor and may form an ionic or hydrogen bond interaction with the primary ammonium group of 5-HT.

Animals↗
PubMedDate not supplied

Detection of implanted metallic devices by airport security.

PURPOSE: Permanent metallic implants are commonly placed by interventional radiologists. The authors evaluate the ability of these and various orthopedic devices to be detected by airport security metal detectors. MATERIALS AND METHODS: A variety of commonly placed radiologic and orthopedic implants were evaluated by three types of metal detectors at Toronto International Airport. RESULTS: No radiologic devices were detected by walk-through detectors. Metal-containing ports were the only device to set off the hand-held scanners. CONCLUSION: Radiology patients can be reassured that their devices will not set off walk-through airport security metal detectors, however, some metallic ports may be detected by hand-held scanning devices.

Aviation↗
PubMedDate not supplied

Stimulus dependence of orientation and direction sensitivity of cat LGNd relay cells without cortical inputs: a comparison with area 17 cells.

The cortical contribution to the orientation and direction sensitivity of LGNd relay cells was investigated by recording the responses of relay cells to drifting sinusoidal gratings of varying spatial frequencies, moving bars, and moving spots in cats in which the visual cortex (areas 17, 18, 19, and LS) was ablated. For comparison, the spatial-frequency dependence of orientation and direction tuning of striate cortical cells was investigated employing the same quantitative techniques used to test LGNd cells. There are no significant differences in the orientation and direction tuning to relay cells in the LGNd of normal and decorticate cats. The orientation and direction sensitivities of cortical cells are dependent on stimulus parameters in a fashion qualitatively similar to that of LGNd cells. The differences in the spatial-frequency bandwidths of LGNd cells and cortical cells may explain many of their differences in orientation and direction tuning. Although factors beyond narrowness of spatial-frequency tuning must exist to account for the much stronger orientation and direction preferences of cells in area 17 when compared to LGNd cells, the evidence suggests that the orientation and direction biases present in the afferents to the visual cortex may contribute to the orientation and direction selectivities found in cortical cells.

Animals↗
PubMedDate not supplied

The renal expression of heat shock protein 47 and collagens in acute and chronic experimental diabetes in rats.

Glomerulosclerosis and tubulointerstitial fibrosis are the main structural changes found in the later stages of diabetic nephropathy, which is clinically characterized by proteinuria, and progressive renal insufficiency. Heat shock protein (HSP) 47, a collagen-binding stress protein, has a specific role in the intracellular processing of procollagen molecules during collagen synthesis. It is implicated in the pathogenesis of various fibrotic diseases. However, the expression and significance of HSP47 in acute and chronic phases of diabetic nephropathy is not yet known. In this study, we studied the expression of HSP47 in the kidneys obtained from streptozotocin-induced diabetic rats, in both short- and long-term diabetes. To determine the renal expression of HSP47, and collagens (type III and IV) in acute (days 1, 3 and 14) and chronic (weeks 4, 12 and 24) diabetes, we have performed a time-course study using streptozotocin-induced diabetic rats. The expression pattern of alpha-smooth muscle actin (to identify mesangial cell damage), vimentin (to identify tubular epithelial cell damage), and desmin (to identify glomerular epithelial cell damage) was also determined in kidneys of these diabetic rats. Antibodies specific for HSP47, type III and type IV collagens, alpha-smooth muscle actin, vimentin, and desmin were used to assess the relative expression of their proteins in paraffin-embedded kidney sections by immunohistochemistry. Compared to control rat kidneys, no significant changes in the expression of HSP47 was found in the kidneys of acute diabetic rats. However a significant increase in the expression of HSP47 was noted in the kidneys of chronic diabetic rats; increased expression of HSP47 correlated with an increased renal deposition of types III and IV collagens. Similarly, compared to kidneys of control and acute diabetic rats, an increased expression of alpha-smooth muscle actin (in mesangial cells), vimentin (in tubular epithelial cells), and desmin (in glomerular epithelial cells) was detected in the kidneys of chronic diabetic rats; by dual immunostaining, these phenotypically-altered renal cells in kidneys of chronic diabetic rats were found to be HSP47-producing cells. Importantly, HSP47 up-regulation coincided with the initiation and progression of renal fibrosis, as determined by the expression and deposition of collagens. Our results strongly support a pathological role for HSP47 in the later stages (sclerotic phase) of streptozotocin-induced diabetic nephropathy, which is associated with glomerulosclerosis and tubulointerstitial fibrosis.

Actins↗