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Biomedical subjects

Z Liu

Publications and source records attributed to Z Liu.

At least 181 records · Page 10Linked to original sources
PubMed2001

Motor neurons rapidly accumulate DNA single-strand breaks after in vitro exposure to nitric oxide and peroxynitrite and in vivo axotomy.

The mechanisms of neuronal degeneration in motor neuron disease are not fully understood. We tested the hypothesis that oxidative stress in vitro and axotomy in vivo induce single-strand breaks (SSB) in DNA, a form of early DNA damage, in adult motor neurons early during their degeneration. We developed and characterized a novel cell suspension system enriched in motor neurons from adult rat spinal cord ventral horn. This cell system is approximately 84% neurons, with approximately 86% of these neurons being motor neurons; approximately 72% of these motor neurons are alpha-motor neurons. Motor neuron viability in suspension is approximately 100% immediately after isolation and approximately 61% after 12 hours of incubation. During incubation, isolated motor neurons generate high levels of superoxide. We used single-cell gel electrophoresis (comet assay) to detect DNA-SSB in motor neurons. Exposure of motor neurons to nitric oxide (NO) donors (sodium nitroprusside or NONOate), H2O2, or NO donor plus H2O2 rapidly induces DNA-SSB and causes motor neuron degeneration, the occurrence of which is dose and time related, as represented by comet formation and cell loss. Motor neuron toxicity is potentiated by cotreatment with NO donor and H2O2 (at nontoxic concentrations alone). Peroxynitrite causes DNA-SSB in motor neurons. The DNA damage profiles (shown by the comet morphology and moment) of NO donors, NO donor plus H2O2, and peroxynitrite are similar. In an in vivo model of motor neuron apoptosis, DNA-SSB accumulate slowly in avulsed motor neurons before apoptotic nuclear features emerge, and the comet fingerprint is similar to NO toxicity. We conclude that motor neurons challenged by oxidative stress and axotomy accumulate DNA-SSB early in their degeneration and that the formation of peroxynitrite is involved in the mechanisms.

Animals↗
PubMed2001

Orphanin FQ: an endogenous antagonist of rat brain dopamine transporter.

Orphanin FQ, also known as nociceptin (NC),is a well-known ligand for opioid receptor-like ORLI receptor. This heptadecapeptide was identified as potently inhibiting the uptake of rat dopamine transporter (rDAT) which is stably expressed in CHO cells (designated D8 cells). Further kinetic analysis proved that this occurs through competitive inhibition with an IC50 of about 1.9 microM. Orphanin FQ also inhibits [3H]dopamine uptake by rat striatal synaptosomes, which confirmed the effect of orphanin FQ on D8 cells. Orphanin FQ was also found to inhibit GABA transporter type I (GATI) but not the serotonin transporter. These results suggest that orphanin FQ is an endogenous antagonist of dopamine transport and that it affects locomotion and other activities at least partly by inhibiting dopamine transporter and directly affecting dopamine transmission or by inhibiting GABA transporter to indirectly change dopaimne transmission.

Animals↗
PubMed2001

Molecular cloning of a cDNA for rat TM4SF4, a homolog of human il-TMP (TM4SF4), and enhanced expression of the corresponding gene in regenerating rat liver(1).

il-TMP (also known as TM4SF4) is a human tetraspanin that is expressed in human intestine and liver. We have cloned a novel cDNA for a rat gene with sequence similar to that of a cDNA for human il-TMP. The cDNA encoded a protein of 202 amino acids, designated rat TM4SF4. The corresponding transcript was detected in rat liver and testis. The expression of rat TM4SF4 was enhanced in regenerating liver after two-thirds partial hepatectomy. It was supposed that rat TM4SF4 might play a role in cell proliferation and in liver regeneration.

Amino Acid Sequence↗
PubMed2001

Application of ab initio molecular dynamics for a priori elucidation of the mechanism in unimolecular decomposition: the case of 5-nitro-2,4-dihydro-3H-1,2,4-triazol-3-one (NTO).

We have tested a new and general approach for the theoretical study of unimolecular decomposition. By combining the power of the ab initio molecular dynamics (MD) and ab initio molecular orbital (MO) methods, our approach requires no prior experimental knowledge or intuitive assumptions about the decomposition. Instead, the reaction channels are first sampled theoretically by simulating a molecule at high temperature in a number of trajectories, using the density functional theory (DFT) based ab initio MD method with a planewave basis set and pseudopotentials. Each type of these channels is then further examined by well-established ab initio MO method to locate the energy barrier and transition structure and to verify the ab initio MD results. The power of such an approach is demonstrated in a case study for the complicated unimolecular thermal decomposition of NTO (5-nitro-2,4-dihydro-3H-1,2,4-triazol-3-one), with several interesting new features uncovered. The C-NO2 homolysis is indeed the dominant channel at high temperature, while the departing NO2 could capture a H atom from the NTO ring to form HONO, by either a concerted bond breaking mechanism or by a bimolecular reaction between the NO2 group and the triazol ring. At lower temperature, the dissociation channels initiated by hydrogen migrations should be activated first. The channel with hydrogen migration followed by ring opening and then by HONO loss has an energy barrier of 38.0 kcal/mol at the rate-determining step, being the lowest among all the investigated dissociation paths and much lower than previously thought. The energy barrier for nitro-nitrite rearrangement is lower than that for the C-NO2 homolysis but makes only a minor contribution due to the entropy factor. And the NTO ring could rupture in the two C-N bonds connected to the carbonyl carbon, and the energy barriers for such processes are only 2-4 kcal/mol higher than that for the C-NO2 homolysis.

Journal Article↗
PubMed2001

Schiff base complexes of vanadium(III, IV, V) as catalysts for the electroreduction of O2 to H2O in acetonitrile.

Fifteen Schiff base ligands were synthesized and used to form complexes with vanadium in oxidation states III, IV, and V. Electrochemical and spectral characteristics of the complexes were evaluated and compared. In acidified solutions in acetonitrile the vanadium(IV) complexes undergo reversible disproportionation to form V(III) and V(V) complexes. With several of the ligands the V(III) complexes are much more stable in the presence of acid than is the previously studied complex with salen, an unelaborated Schiff base ligand (H(2) salen = N,N'-ethylenebis(salicylideneamine)). Equilibrium constants for the disproportionation were evaluated. The vanadium(III) complexes reduce dioxygen to form two oxo ligands. The reaction is stoichiometric in the absence of acid, and second-order rate constants were evaluated. In the presence of acid some of the complexes investigated participate in a catalytic electroreduction of dioxygen.

Journal Article↗
PubMed2001

Isolation and characterization of mouse MUC18 cDNA gene, and correlation of MUC18 expression in mouse melanoma cell lines with metastatic ability.

The cell surface adhesion molecule human MUC18 (huMUC18 or Mel-CAM) has been postulated to play a key pathogenic role in metastatic melanoma progression. To establish an immunocompetent syngeneic mouse model that would greatly facilitate our understanding of the role of MUC18 in the metastatic behavior of melanoma, we cloned and characterized the mouse MUC18 (muMUC18) cDNA gene. The gene was amplified by RT-PCR and RACE of the poly(A)+RNA isolated from the mouse melanoma cell line B16F10/Queens. The cloned muMUC18 cDNA gene contained 28 nucleotides of 5'-UTR, 908 nucleotides of 3'-UTR, and an open reading frame (ORF) of 1947 nucleotides encoding a protein of 648 amino acids, which is two amino acids longer than huMUC18. The size of the muMUC18 mRNA is about 3 kb with a shorter 3'-UTR than the huMUC18 mRNA (about 3.3 kb). Besides, the sequence in the 3' UTR of the two mRNAs is diverse with only 31% identity. The 5'-UTR and coding sequences of the muMUC18 cDNA are 72.4 and 80.6% identical to those of huMUC18, respectively. The deduced amino acid sequence of the muMUC18 cDNA is 76.2% identical to that of huMUC18. The amino acid sequences deduced from MUC18 cDNA sequences from six other mouse melanoma cell lines are identical except one to three residues, suggesting that the muMUC18 cDNA sequence determined in this report is correct. The muMUC18 protein is predicted to be slightly more acidic than the human protein. The levels of muMUC18 mRNA and protein in nine mouse melanoma cell lines were directly proportional to their ability to establish metastatic colonies in lungs of syngeneic mice. Most biological functions of the muMUC18 may be similar to the huMUC18.

Amino Acid Sequence↗
PubMed2001

Expression and activity of matrix metalloproteases in human malignant mesothelioma cell lines.

The extracellular matrix metalloproteases (MMPs) secreted by various human tumor cells play a crucial role in tumor cell invasion and metastasis, but their expression in malignant mesothelioma (MM) cells has not been examined. In this study, we have investigated the spectrum of MMPs and tissue inhibitors of metalloproteases (TIMPs) produced by 8 MM cell lines. Using RT-PCR, we found that all investigated MM cell lines expressed genes encoding mRNA for MMP-1 (interstitial collagenase), MMP-2 (gelatinase A), MMP-3 (stromelysin-1), MMP-9 (gelatinase B) and TIMPs 1, 2 and 3. We also found that 6/8 MM cell lines expressed MMP-7 (matrilysin) and 3/8 MM cell lines expressed MMP-10 (stromelysin-2). MMP-11 (stromelysin-3) was not detected in any of the MM cell lines. Production of MMP-2 and MMP-9 was confirmed using gelatin zymography. In addition, all MM cell lines secreted a 66 kDa metalloprotease, while 3/8 MM cell lines secreted 46, 48, 51 and 63 kDa metalloproteases which specifically degraded the extracellular matrix components fibronectin, vitronectin and laminin. The 66 kDa protease was identified as MMP-3 by Western blot. Our results reveal a broad spectrum of MMPs and TIMPs produced by MM cells and indicate that different substrate specificities of MMPs may play a role in MM cell invasion.

Blotting, Western↗
PubMed2001

Analysis of gap junction formation in rat hepatocytes by intravenous injection of an anti-connexin32 monoclonal antibody (HAM8).

HAM8 monoclonal antibody was used to examine the mechanism of connexin32 (Cx32) formation in the rat model in vivo by immunofluorescence and immunoelectron microscopy. After a single intravenous injection of HAM8 IgG, a number of HAM8 signals were observed at the sinusoidal face, between adjacent hepatocytes as well as in the cytoplasm stained with only 2nd fluorescent antibody. Moreover, the in vivo localization of the HAM8 antigen appeared to change with time. At 5 min after the antibody injection, Cx32 signals from liver sections were clearly detected at the sinusoidal face. Fifteen minutes later, numerous linear and dotted fluorescent signals were observed between hepatocytes; in addition, much punctate staining was found at the sinusoidal face and in hepatocytes. These findings were identified by immunoelectron microscopy. Interestingly, 1 hr later, much punctate staining considered to be similar to those seen in the normal rat liver tissues was observed between adjacent hepatocytes, suggesting that a great deal of Cx32 combined with HAM8 have been assembled into identifiable gap junction plaques. Five hours later, intercellular and intracellular Cx32 signals were infrequently detected. When staining was performed with HAM8 and 2nd antibody, however, numerous Cx32 signals were again observed between neighboring hepatocytes, as punctate staining appearing in a pattern approximately the same as that seen in the normal liver tissue. Based on these results, we assumed that a precursor gap was present during Cx32 formation, and discussed the pathways of Cx32 formation and the degradation of Cx32 as well as that of HAM8.

Animals↗
PubMed2001

Gene expression profile changes in initiation and progression of squamous cell carcinoma of esophagus.

Tumorigenesis is a complex process involving multiple genes. As a step toward understanding the complicated changes between normal and malignant cells, this report focused on gene expression profile variations among normal and abnormal esophageal epithelium tissues. The cDNA microarray approach was used to investigate gene expression profiles of 5 different stages during initiation and progression of esophageal cancer. According to pathological characteristics, these 5 stages were normal, dysplasia I (mild dysplasia), dysplasia II (moderate dysplasia), carcinoma in situ (CIS) and squamous cell carcinoma of esophagus (SCC). Comparing and analyzing those gene expression profiles, we observed that the expression levels of many genes changed in dysplasia I and some known tumor-related genes were over-expressed or under-expressed in all 4 abnormal stages. Using principle component analysis we identified a set of genes that may play an important role in tumor development. Hybridization data were confirmed by semi-quantitative reverse transcription-polymerase chain reaction and immunohistochemistry. These results suggest that cDNA microarray technology is a useful tool to discover genes frequently involved in esophageal neoplasia and provides novel clues to diagnosis, early detection and intervention of SCC.

Carcinoma in Situ↗
PubMed2001

Mouse model of congenital polycythemia: Homologous replacement of murine gene by mutant human erythropoietin receptor gene.

Mutations causing truncations of the cytoplasmic domain of the human erythropoietin receptor (EPOR) result in a dominantly inherited disorder-primary familial congenital polycythemia. This disorder is characterized by increased numbers of erythrocytes (polycythemia) and by in vitro hypersensitivity of erythroid precursors to erythropoietin. The consequences of EPOR truncation in nonerythroid tissues are unknown. We replaced the murine EPOR gene with a wild-type human EPOR gene and a mutant human EPOR gene that we initially identified in a patient with polycythemia. This mutation leads to an EPOR truncated after the first tyrosine residue of the intracellular domain. Mice heterozygous for this mutant allele and a wild-type human EPOR allele mimicked the human disorder. Interestingly, mice that were homozygous for the mutant human allele were severely polycythemic but viable. Our results provide a model for functional studies of EPOR-triggered signaling pathways in erythropoiesis. These animals can now be used to investigate the molecular pathophysiology of this gain-of-function EPOR mutation in erythroid tissue and in those nonerythroid tissues that express EPOR.

Animals↗
PubMed2001

Detection of Yersinia ruckeri in rainbow trout blood by use of the polymerase chain reaction.

We evaluated a polymerase chain reaction (PCR) method for detecting Yersinia ruckeri, the bacterial pathogen causing enteric redmouth disease (ERM), in blood of rainbow trout Oncorhynchus mykiss. Identification of the PCR product was confirmed by Southern blot hybridization with a 32P-labeled oligonucleotide probe matching a sequence within the small subunit ribosomal RNA gene of Y. ruckeri. Following a 1 h immersion of rainbow trout in water with 4.5 x 10(6) colony-forming units of Y. ruckeri l(-1), the PCR was positive for all blood samples from 1 h (first sample) to 5 d and was negative from 9 to 30 d (last sample). Fish in this experiment did not show signs of disease, probably because they had been vaccinated against Y. ruckeri. To test this method with naturally infected fish, 42 rainbow trout from hatcheries were examined. Four of these fish had clinical signs of ERM and were infected with Y. ruckeri based on bacteriological culture. The PCR method detected Y. ruckeri in blood, intestine, liver, and trunk kidney from the 4 fish with ERM and from 5 additional rainbow trout that were bacteriologically negative for Y. ruckeri. Three of 5 rainbow trout from streams receiving effluent from hatcheries were positive for Y. ruckeri when tested with PCR, although there was no growth of Y. ruckeri on culture plates inoculated with the same samples. Samples were successfully stored for 1 wk in lysis buffer at 25 degrees C. This study demonstrated that a non-lethal blood sample can be used with PCR to detect Y. ruckeri.

Animals↗
PubMed2001

[Combination treatment for craniopharyngiomas with stereotactic intracavitary irradiation and gamma knife surgery].

OBJECTIVE: To evaluate the effectiveness of the combination treatment for craniopharygiomas with stereotactic intracavitary irradiation and gamma knife surgery. METHODS: Combination treatment of stereotactic instillation of radioisotopes and gamma knife surgery was performed in 46 patients with craniopharyngioma between October 1996 and June 1999. There were 24 men and 22 women, with age ranging from 3 to 60 years (average 38.6 years). There were 13 solid tumors and 33 mixed solid and cystic tumors. Twenty-eight patients had undergone prior caraniotomies with total or partial resection of tumors, with or without followed external beam radiation therapy. Gamma knife surgery was done in solid tumors and the combination treatment was done in mixed tumors: the cyst being treated with stereotactic intracavitary radiation and the solid part with Gamma knife. RESULTS: Thirty-eight patients (10 solid and 28 mixed) were followed up from 6 months to two years (averaged 16 months). The tumor control rate (disappeared, decreased and unchanged) was 90% in solid tumor, 85.7% in mixed tumors, and 92.1% in solid segment and the total control rate was 89.5%. CONCLUSION: The combination treatment is a simple, safe and effective method for craniopharyngiomas, especially for the recurrent mixed solid and cystic tumors. Gamma knife surgery plays an important role in the treatment of solid component of the craniopharyngiomas.

Adolescent↗
PubMed2001

Cloning and characterization of full-length human ribosomal protein L15 cDNA which was overexpressed in esophageal cancer.

The aim of this investigation was trying to identify the genes differentially expressed in esophageal cancer. By combining suppression subtractive hybridization (SSH) with reverse Northern high density blots, a gene named EC45 was obtained, which dramatically overexpressed in 70% esophageal cancer (18/26). EC45 was mapped to 3p12-3p11.2 by radiation hybrid mapping (RH mapping). The putative full length EC45 cDNA (1987 bp) was identified by cDNA libraries screening of esophageal cancer. EC45 encoded 204 amino acids, and it shared a 100% similarity with ribosomal protein L15 (635 bp, mRNA) in ORF, but no similarity in 5' UTR or 3' UTR. Northern blot panel of multiple adult human normal tissues showed EC45 distributed in almost normal tissues tested. All these data suggested that EC45, encoding ribosomal protein L15 and overexpressing in esophageal cancer might play a possible role in carcinogenesis of esophagus.

Amino Acid Sequence↗
PubMed2001

Rh type B glycoprotein is a new member of the Rh superfamily and a putative ammonia transporter in mammals.

Ammonium transporters play a key functional role in nitrogen uptake and assimilation in microorganisms and plants; however, little is known about their structural counterpart in mammals. Here, we report the molecular cloning and biochemical characterization of Rh type B glycoproteins, human RhBG and mouse Rhbg, two new members of the Rh family with distinct tissue specificities. The RhBG orthologues possess a conserved 12-transmembrane topology and most resemble bacterial and archaeal ammonium transporters. Human RHBG resides at chromosome 1q21.3, which harbors candidate genes for medullary cystic kidney disease, whereas mouse Rhbg is syntenic on chromosome 3. Northern blot and in situ hybridization revealed that RHBG and Rhbg are predominantly expressed in liver, kidney, and skin, the specialized organs involving ammonia genesis, excretion, or secretion. Confocal microscopy showed that RhBG is located in the plasma membrane and in some intracellular granules. Western blots of membrane proteins from stable HEK293 cells and from mouse kidney and liver confirmed this distribution. N-Glycanase digestion showed that RhBG/Rhbg has a carbohydrate moiety probably attached at the NHS motif on exoloop 1. Phylogenetic clustering, tissue-specific expression, and plasma membrane location suggest that RhBG homologous proteins are the long sought major ammonium transporters in mammalians.

Amino Acid Sequence↗
PubMed2001

Sequential immunization of macaques with two differentially attenuated vaccines induced long-term virus-specific immune responses and conferred protection against AIDS caused by heterologous simian human immunodeficiency Virus (SHIV(89.6)P).

Four rhesus macaques were sequentially immunized with live vaccines DeltavpuDeltanefSHIV-4 (vaccine-I) and Deltavpu SHIV(PPC) (vaccine-II). The vaccine viruses did not replicate productively in the peripheral blood mononuclear cells (PBMCs) of the vaccinated animals. All four animals developed binding antibodies against both the vaccine-I and -II envelope glycoproteins but neutralizing antibodies only against vaccine-I. They developed vaccine virus-specific CTLs that also recognized homologous as well as heterologous pathogenic SHIVs. Thirty weeks after the last immunization, the vaccinated animals and three unvaccinated control animals were challenged iv with a highly virulent heterologous SHIV(89.6)P. As expected, the three unvaccinated control animals developed large numbers of infectious PBMCs, high plasma viremia, and precipitous loss of CD4(+) T cells. Two controls did not develop any immune response and succumbed to AIDS in about 6 months. The third control animal developed neutralizing antibodies and had a more chronic disease course, but eventually succumbed to AIDS-related complications 81 weeks after inoculation. The four vaccinated animals became infected with challenge virus as indicated by the presence of challenge virus-specific DNA in the PBMCs and RNA in plasma. However, virus in these animals replicated approximately 200- to 60,000-fold less efficiently than in control animals and eventually, plasma viral RNA became undetectable in three of the four vaccinates. The animals maintained normal CD4(+) T-cell levels throughout the observation period of 85 weeks after a transient drop at Week 3 postchallenge. They also maintained CTL responses throughout the observation period. These studies thus showed that the graded immunization schedule resulted in a safe and highly effective long-lasting immune response that was associated with protection against AIDS by highly pathogenic heterologous SHIV(89.6)P.

AIDS Vaccines↗
PubMed2001

Modeling the third loop of short-chain snake venom neurotoxins: roles of the short-range and long-range interactions.

The influence of long-range interactions on local structures is an important issue in understanding protein folding process and protein structure stability. Using short-chain snake venom neurotoxin as a model system, we have studied the conformational properties of eight different loop III sequences either in the environment of one of the short-chain neurotoxin, erabutoxin b (PDB ID 1nxb), or in free state by Monte Carlo simulated annealing method. The surrounding protein structure was found to be crucial in stabilizing the loop conformation. Although all the eight peptides prefer type V beta turn in solution, three of them (KPGI, KPGV, KSGI) turn to type II beta turn and the other five (KKGI, KKGV, KNGI, KQGI, and KRGV) are confined to more rigid type V beta turn conformation in the protein structure. Using flexible tetra-glycine-peptide to screen the backbone conformational space in the protein environment also validates the results. This study shows that long-range interactions do contribute to the stability and the types of conformation for a surface loop in protein, while short-range interactions may only provide candidate conformations, which then have to be filtered by the long-range interactions further.

Amino Acid Motifs↗