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Biomedical subjects

Z Liu

Publications and source records attributed to Z Liu.

At least 1,207 records · Page 67Linked to original sources

Changes in nuclear size of cardiac myocytes during the development and progression of hypertrophy in rats.

It is generally believed that nuclear enlargement indicates polyploidy. The purpose of this study was to establish whether nuclear enlargement is also a marker for cellular hypertrophy. Using isolated myocytes, we examined the growth of cardiac myocyte nuclei during cellular hypertrophy in rats with aortocaval fistulas or left ventricular myocardial infarction. A Coulter Channelyzer was used to measure the volume of the myocytes. Isolated myocytes were stained with the DNA-specific fluorochrome 4'-6-di-amidino-2 phenylindole-HCl for measurements of nuclear length and width, and calculation of nuclear volume. One week, 1 month and 5 months after aortocaval fistula surgery, the nuclear volume of right ventricular myocytes increased by 24, 55 and 56% respectively. Increased length, rather than width, accounted for most of the nuclear growth. Nuclear hypertrophy was associated with a progressive increase in cell volume at each time point (34, 88 and 118%). Adaptive growth of left ventricular myocytes followed the same trend, though the extent of cellular and nuclear hypertrophy was reduced. One month after producing a myocardial infarction, there was an increase in nuclear volume (18%) and nuclear length (11%) in right ventricular myocytes, but no changes in the surviving left ventricular myocytes. The cell volume increased in both right and left ventricles (72 and 18%, respectively). Thus, nuclear size increased as myocytes enlarged, though at a slower rate. Since nuclear DNA content does not increase in rats with aortocaval fistulas or myocardial infarction, the increase in nuclear volume was associated with cellular enlargement rather than increased polyploidy.

Animals↗

[Corneal topography of patients with unsatisfactory visual acuity after epikeratophakia for aphakia].

Corneal topographic characteristics of 5 patients who obtained unsatisfactory vision after aphakic epikeratophakia, with at least nine months follow-up, were reported. All grafts were apparently decentered relative to visual axis. The contour of cornea showed highly irregular pattern of power distribution. These findings suggest that to mark visual axis before surgery, lathe corneal lenses carefully and do standard operation should be the basis of successful epikeratophakia.

Adolescent↗

[Amplification in vitro and identification of small subunit ribosomal DNA fragment of Plasmodium falciparum].

According to computer analysis of SSUrDNA sequences of Plasmodium, other protozooa and human, two oligonucleotide primers were designed. A DNA fragment, about 570 base pairs, was successfully amplified by two temperature point polymerase chain reaction from the genomic DNA of cultivated erythrocytic stage of P. falciparum FCC/YN (Simao), but no fragment was obtained from that of P. vivax, L. donovani, T. gondii and humans. It has been confirmed that the amplified fragment was indeed expected SSUrDNA segment of P. falciparum by means of restriction endonuclease digestions and Northern blot hybridization.

Animals↗

Resistance to HIV-1 infection. Multicenter AIDS Cohort Study.

Men from the Multicenter AIDS Cohort Study were classified as "susceptible" and "resistant" to HIV infection. Resistant men were still HIV antibody negative in 1993 and were estimated to have had > 45 different anal intercourse partners (median, 92; range, 46-504) in the 2.5 years before visit 2 (1985). Susceptible men were seroconverters who were estimated to have had < 13 different anal partners (median, 4; range, 0-12). Leukocyte groups were compared between the two groups of men. Values were excluded for 12 months before the first antibody-positive visit in the susceptible men. White blood cells, polymorphonuclear neutrophils, total lymphocyte count, CD8+ percentage and number, and CD3+ and CD4+ number were higher in the resistant men. Logistic regression analyses were used to develop 50 bivariate models. Higher levels of neutrophils and CD8+ cells were included in four of the six best-fitting bivariate models, suggesting that each is associated with resistance to HIV-1 infection. These results support the hypothesis that CD8+ cells may modulate the outcome of HIV-1 exposure.

CD8-Positive T-Lymphocytes↗

High-level expression of staphylococcal nuclease A in Escherichia coli.

The staphylococcal nuclease A gene has been successfully cloned and overexpressed in E. coli under the transcriptional control of the bacteriophage lambda PRPL promoters regulated by the temperature sensitive repressors. The SDS-PAGE analysis demonstrates that the nuclease A is produced to the extent of as much as 60% of the total cellular protein. The N-terminal analysis of the nuclease A shows that the amino terminal formyl methionine residue of the enzyme is precisely processed. The recombinant nuclease A with full activity is finally obtained after appropriate solubilization--denaturation and renaturation treatment. The conformational identity of the renatured nuclease A in different conditions is also studied by using hydrophobic interaction chromatography on a phenyl-superose HR5/5 column.

Base Sequence↗

Clinical observation on osteoporosis treated with traditional kidney-tonifying medicaments.

The method of scoring kidney deficiency symptoms and bone density was used to judge therapeutic efficacy on osteoporosis of traditional kidney-tonifying drugs. It was proved that the drugs not only improved clinical symptoms but also delayed the developmental process of this disorder. It also verified the correctness of the traditional theory that the kidney dominates the bone and generates the marrow.

Adult↗

Effect of temperature on kainic acid-induced seizures.

The effects of body temperature on kainic acid-induced seizures and seizure-related brain damage were examined in rats. In rats with status epilepticus induced by intraperitoneal injection of 12 mg/kg of kainic acid (KA), ictal discharges were decreased by 50% when body temperature was lowered to 28 degrees C and nearly abolished when body temperature was lowered to 23 degrees C. In rats with mild hypothermia (28 degrees C), the duration of ictal discharges following KA injection was significantly lower than in rats with normal body temperature. No detectable hippocampal cell loss was observed in rats with hypothermia to 28 degrees C whereas gross cell loss in the hippocampus was observed in all rats with KA injection at normal body temperature. In contract to hypothermia, hyperthermia markedly aggravated the seizures and hippocampal damage induced by KA. Following elevation of body temperature to 42 degrees C KA (12 mg/kg) resulted in severe seizures and all rats died of tonic seizures within 2 h. Furthermore, 6 mg/kg of KA administered to rats with a body temperature of 41-42 degrees C, resulted in up to 4 h of continuous ictal discharges whereas no continuous ictal discharges were observed after the same injections in rats with normal body temperature. Histological examination in rats receiving 6 mg/kg of KA revealed severe cell loss in the hippocampus in rats with hyperthermia but not in rats with normal temperature. These results demonstrate that body temperature plays an important role in the control of epileptic seizures and seizure-related brain damage.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of nonionic detergents on P-glycoprotein drug binding and reversal of multidrug resistance.

Multidrug-resistant cells are thought to maintain low intracellular cytotoxic drug concentration though the active efflux of drugs across the cell membrane. It is presently believed that P-glycoprotein mediates this energy-dependent drug efflux by interacting directly with various lipophilic compounds. In this report, we have used [3H]azidopine in a photoaffinity labeling assay to study the effect of detergents and denaturing agents on P-glycoprotein drug binding in intact cells. Nonionic detergents such as Triton X-100 or Nonidet P-40 at very low concentrations were found to completely abolish azidopine photolabeling to P-glycoprotein and are able to reverse the multidrug resistance phenotype. In contrast, high concentrations of the denaturing agent urea or the zwitterionic detergent 1-[(3-cholamidopropyl)dimethylamino]-1-propanesulfonate did not inhibit azidopine photolabeling to P-glycoprotein. A comparison between verapamil and Triton X-100 revealed that the latter was more effective in inhibiting azidopine photolabeling to P-glycoprotein while verapamil was more effective in potentiating [3H]vinblastine accumulation in drug-resistant cells. Drug transport studies showed that [3H]Triton X-100 accumulated in both drug-sensitive and -resistant cells, and its accumulation was not modulated by excess vinblastine, verapamil, or colchicine. Taken together, these findings suggest that low concentrations of Triton X-100 reverse the multidrug resistance phenotype by inhibiting P-glycoprotein drug binding. In addition, it is also suggested that the site(s) of P-glycoprotein drug binding is localized to sequences found within the lipid bilayer.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Naturally processed HLA class I bound peptides from c-myc-transfected cells reveal allele-specific motifs.

Naturally processed peptides, bound to HLA-A2, A68, B40 molecules, were isolated from a c-myc transfected lymphoblastoid B cell lines for sequence analysis. Forty-three sequences of bound peptides could be grouped into three structural motifs. One of the peptide sequences obtained, SLLPAIVEL, was identical to a previously reported peptide bound to HLA-A2.1 and was used for grouping HLA-A2-bound peptides. A second motif, identical to that previously reported for HLA-A68-bound peptides, was also observed. A distinct third motif, consistent with the structure of the HLA-B40 "45 pocket," was observed. The peptides within this group contained glutamate in position 2, usually followed by a hydrophobic residue in positions 3 and 9. Within this motif group of peptides bound to MHC class I molecules, one peptide, HEETPPTTS, was 100% homologous to residues 243-251 of the c-myc protein.

Actins↗

Naturally processed cytokine-derived peptide bound to HLA-class II molecules.

Sequence analysis of HLA-class II (HLA-DR beta 1-1502 and 1104)-bound self-peptides from a transformed B cell line was performed. The sequences of naturally processed self-peptides bound to HLA-DR2 and DR5 were compared with protein and nucleic acid data bases for homology to known precursor proteins. Of the matches to known precursors, one peptide showed 100% homology to the third framework and CDR3 regions of Ig VH expressed by the line. Another peptide matched 100% to the human equivalent of macrophage inflammatory protein (MIP). A synthetic peptide corresponding to the naturally processed form of MIP (KPGVIFLTKRSRQV) was shown to inhibit Ag-specific HLA-DR beta 1*1104-restricted T cell proliferation. This indicates that the MIP peptide binds to HLA-DR beta 1*1104. The MIP peptide belongs to a set of peptides that showed uniform NH2-terminal processing. In this set, proline always occurred as the second residue followed by a basic lysine or arginine in position nine. This suggests that final NH2-terminal processing of peptides precedes their binding to MHC molecules. A distinct, second set of peptides showed ragged NH2-terminii, as has been reported for other naturally processed MHC-class II-bound self-peptides.

Amino Acid Sequence↗

RPII15 codes for the M(r) 15,000 subunit 9 of Drosophila melanogaster RNA polymerase II.

The RPII15 gene product of Drosophila melanogaster, which has recently been identified by sequence comparison, possesses a high similarity to subunit 9 of yeast RNA polymerase II. Using the polymerase chain reaction the coding region of RPII15 was isolated from genomic DNA of adult flies. Sequence analysis shows four amino acid substitutions in comparison to the previously reported sequence. Antisera were generated against bacterially expressed RPII15 and were used for immunoblotting experiments with RNA polymerase II of Drosophila melanogaster. This analysis identified the M(r) 15,000 subunit 9 as gene product of RPII15.

Amino Acid Sequence↗

Bullous pemphigoid and herpes gestationis autoantibodies recognize a common non-collagenous site on the BP180 ectodomain.

Bullous pemphigoid (BP) and herpes gestationis (HG) are skin diseases characterized by subepidermal blisters and autoantibodies against two hemidesmosomal Ag, i.e., BP230 and BP180. Based on sequence analysis the BP180 Ag was predicted to be a transmembrane protein with a long extracellular collagenous domain. In the present investigation fusion proteins encompassing various segments of the BP180 Ag were expressed in a prokaryotic system and assayed by immunoblotting and immunoadsorption against a panel of BP, HG and control sera. One antigenic site, comprising 14 amino acids of the BP180 noncollagenous (NC) 16A domain, was shown to be recognized by 60% of BP sera and by 63% of HG sera tested. 73% (11/15) of BP sera and 100% (8/8) of HG sera reacted with at least one of three BP180 fusion proteins representing various portions of the NC16A domain. Immunoadsorption analysis identified this region of BP180 as an immunodominant site. Using an affinity purified rabbit antiserum raised against a recombinant form of BP180, this BP/HG autoantibody-reactive region was localized to the epidermal basal lamina immediately adjacent to the hemidesmosome. These findings confirmed the predicted type II transmembrane orientation of the BP180 Ag. Thus, the long, C-terminal collagenous domain of this basal keratinocyte protein projects into the basal lamina and may function as a site of interaction with an extracellular matrix component. It is proposed that autoantibodies directed against the well-defined antigenic site on the BP180 ectodomain may play an initiatory role in subepidermal blister formation in BP and HG patients.

Amino Acid Sequence↗

Neuroprotective effect of chronic infusion of basic fibroblast growth factor on seizure-associated hippocampal damage.

Basic fibroblast growth factor (bFGF) has been shown to have neuroprotective effects in animal models of ischemia. To determine whether bFGF is protective against seizure-induced brain damage, we administered bFGF through osmotic pumps prior to, and after treatment with kainic acid (KA). Recombinant bFGF, CS23, a modified human bFGF, was infused into the lateral ventricles in rats for 2 days before and 5 days after the injection of KA. Control rats received equal volumes of phosphated saline over the same period of time. Infusion of 5 micrograms/ml of bFGF (0.5 microliter/h) did not modify the latency and duration of seizures induced by intraperitoneal injections of KA. However, bFGF prevented cell loss in the hippocampus in 80% of the rats. In control rats, cell loss in the hippocampus was found in all rats. These results indicate that bFGF has a substantial neuroprotective effect.

Animals↗

Effects of quisqualic acid and glutamate on subsequent learning, emotionality, and seizure susceptibility in the immature and mature animal.

To compare the long-term behavioral effects of chronic administration of excitatory amino acids in the mature and immature brain quisqualic acid (QA) and glutamate (GLU) were administered intraventricularly by osmotic pumps over 7 days in 20- and 60-day-old rats. Both age groups received identical dosages of QA or GLU. At age 90 days, all animals were assessed for abnormalities of learning and memory using the Morris water maze, emotionality using the handling test, and seizure threshold using flurothyl inhalation. No significant differences were found in either the water maze or handling test. However, 60-day-old rats receiving QA or GLU had more spontaneous seizures than the 20-day-old rats. In both age groups histological damage following QA and GLU was limited to the ipsilateral hippocampus, was maximum at the site of the catheter tube, and was similar in the two age groups studied. The adverse effects of long-term effects of chronic exposure to excitatory amino acids are similar in the immature and mature brain.

Administration, Inhalation↗

Polyoma virus early-late switch: regulation of late RNA accumulation by DNA replication.

Early in infection of permissive mouse cells, messages from the early region of the polyoma virus genome accumulate preferentially over those from the late region. After initiation of DNA replication, the balance between early and late gene expression is reversed in favor of the late products. In previous work from our laboratory, we showed that viral early proteins do not activate the polyoma late promoter in the absence of DNA replication. Here we show that activation of the late genes in replication-incompetent viral genomes can occur if actively replicating genomes are present in the same cell. A low level of DNA replication, however, is insufficient to induce the early-late switch. Furthermore, replication-competent genomes that fail to accumulate late RNA molecules are defective in the transactivation of replication-incompetent genomes. We suggest that titration of an unknown diffusible factor(s) after DNA replication relieves the block to late RNA accumulation seen in the early phase, with most of this titration being attributable to late-strand RNA molecules themselves.

3T3 Cells↗

An endogenous glycogen-associated compound modulates glucose-6-phosphatase activity in rat liver microsomes.

Liver glycogen is closely associated with the endoplasmic reticulum, which contains the glucose-6-phosphatase enzyme system that catalyses the final step of hepatic glucose production. To examine whether this structural association has functional consequences, microsomes were isolated from 48 h fasted (n = 6) and ad lib fed rats (n = 3). Microsomes from fed rats had a higher glycogen content and lower enzyme activity than fasted rats. Overall, glucose-6-phosphatase activity was inversely proportional to microsomal glycogen content. Partially purified rabbit or rat liver glycogen, at physiological relevant concentrations (10-100 mM glucose equivalents), added directly to either intact or Triton-disrupted microsomes from fasted rats significantly decreased glucose-6-phosphatase activity. Inhibitory activity was present in native liver glycogen prepared by sedimentation and could be dissociated from glycogen by ion-exchange and ultrafiltration. These findings suggest that a low molecular weight (< 5000 D) compound closely associated with glycogen can modulate glucose-6-phosphatase and may have a physiologic role in the regulation of hepatic glucose production.

Animals↗

Expression of beta subunit isoforms of the Na+,K(+)-ATPase is muscle type-specific.

Hindlimb skeletal muscles of the rat express two isoforms of the alpha (alpha 1 and alpha 2) and two isoforms of the beta (beta 1 and beta 2) subunits of the Na+,K(+)-ATPase. Because several muscles constitute the hindlimb, we investigated if specific isoforms are expressed in particular muscles. Northern blot analysis using isoform-specific cDNA probes demonstrated that soleus muscle expressed only the beta 1 transcript, whereas EDL or white gastrocnemius muscles expressed only the beta 2 transcript, and red gastrocnemius muscle expressed both mRNAs. All muscles tested expressed both alpha 1 and alpha 2 transcripts, albeit to various degrees: alpha 1 transcripts were present to about the same extent in all muscles but alpha 2 mRNA was 4-fold more abundant in soleus than in EDL for the same amount of total RNA. Beta subunit protein levels were investigated in purified plasma membrane fractions of pooled red (soleus + red gastrocnemius + red quadriceps) or white (white gastrocnemius + white quadriceps) muscles using isoform-specific antibodies. Red muscles expressed mostly the beta 1 protein while white muscles expressed mostly the beta 2 subunit. Both muscle groups had similar levels of alpha 1 or alpha 2 subunits, and crude membranes isolated from red muscles had 30% higher Na+,K(+)-ATPase activity than white muscle membranes. We conclude that oxidative muscles (slow and fast twitch) express beta 1 subunits, whereas glycolytic, fast twitch muscles express beta 2 subunits, and that both beta isoforms support the Na+,K(+)-ATPase activity of the alpha subunits.

Animals↗

Immunodominance: a single amino acid substitution within an antigenic site alters intramolecular selection of T cell determinants.

The mechanism of immunodominance was studied by mutating a single amino acid residue within an immunodominant determinant of Staphylococcus aureus nuclease (Nase). Residues 81 to 100, which can be further reduced to 86 to 100, were determined to be the immunodominant determinant of Nase in H-2k mice. By introducing selected single amino acid substitutions into the peptide encompassing residues 86 to 100 (p86-100), residue 90 was shown to be one of the critical amino acids for T cell recognition, inasmuch as most of the T cells recognizing p86-100 do not respond to a p86-100 analog with a substitution of leucine for alanine at the residue 90. A mutant of Nase with a replacement of alanine by leucine at residue 90 (A90L) was constructed, and for A90L region 112 to 130, which is a subdominant determinant in Nase, becomes immunodominant. Although unable to respond to Nase, T cells primed in vivo with the peptides covering various cryptic determinants proliferate when challenged with A90L in vitro. Our results suggest that at the protein level there is competition among potential T cell determinants of protein Ag for binding to MHC molecules, and that this competition plays a role in determining which determinant may become immunodominant.

Amino Acid Sequence↗