Search PubMed⌕ Search

Biomedical subjects

Y Wu

Publications and source records attributed to Y Wu.

At least 523 records · Page 29Linked to original sources

Rapid repression of maize invertases by low oxygen. Invertase/sucrose synthase balance, sugar signaling potential, and seedling survival.

We show here that invertase gene expression and the invertase-sucrose (Suc) synthase ratio decrease abruptly in response to low oxygen in maize root tips. In addition to aiding in the conservation of carbon and possibly ATP, this response has the potential to directly affect sugar signaling relative to carbon flux. Experiments were motivated by the potential for a reduced invertase/Suc synthase balance to alter the impact of respiratory and/or membrane carbon flux on sugar signaling. Maize (Zea mays L.) seedlings with 5-cm primary roots were exposed to anoxic (0% [v/v] O2), hypoxic (3% [v/v] O2), and aerobic conditions. Rapid repression of the Ivr1 and Ivr2 maize invertases by low oxygen was evident in root tips within 3 h at both the transcript and activity levels. The speed and extent of this response increased with the degree of oxygen deprivation and differed with genotypes. This decrease in expression also contrasted markedly to that of other genes for respiratory Suc metabolism, such as Suc synthases, which typically increased or remained constant. Although previous work showed that the contrasting effects of sugars on Suc synthase genes were reflected in their regulation by hypoxia and anoxia, the same was not observed for the differentially sugar-responsive invertases. Theoretically advantageous reductions in the invertase/Suc synthase balance thus resulted. However, where this response was extreme (an Oh43 inbred), total sucrolytic capacity dropped below an apparent minimum and root tip viability was reduced. Paradoxically, only Oh43 seedlings showed survival levels >80% (versus <50%) after extreme, long-term stress, suggesting a possible advantage for multiple means of reducing sink activity. Overall, our results demonstrate a rapid change in the regulation and balance of invertases and Suc synthases that could have an immediate impact on limiting the extent of Suc cleavage and reducing the extent of concomitant, hexose-based sugar signaling under low oxygen.

Enzyme Repression↗

Porphyromonas gingivalis virulence factors and invasion of cells of the cardiovascular system.

Our laboratory is interested in the genes and gene products involved in the interactions between Porphyromonas gingivalis (Pg) and the host. These interactions may occur in either the periodontal tissues or other non-oral host tissues such as those of the cardiovascular system. We have previously reported the cloning of several genes encoding hemagglutinins, surface proteins that interact with the host tissues, and are investigating their roles in the disease process. Primary among these is HagA, a very large protein with multiple functional groups that have significant sequence homology to protease genes of this species. Preliminary evidence indicates that an avirulent Salmonella typhimurium strain containing hagA is virulent in mice. These data indicate that HagA may be a key virulence factor of Pg. Additionally, we are investigating the invasion of primary human coronary artery endothelial cells (HCAEC) by Pg because of the recent epidemiological studies indicating a correlation between periodontal disease (PD) and coronary heart disease (CHD). We found that some, but not all, strains of Pg are able to invade these cells. Scanning electron microsopy of the infected HCAEC demonstrated that the invading organisms initially attached to the host cell surface as aggregates and by a "pedestal"-like structure. By transmission electronmicroscopy it could be seen that internalized bacteria were present within multimembranous compartments localized with rough endoplasmic reticulum. In addition, invasion of the HCAEC by Pg resulted in an increase in the degradation of long-lived cellular proteins. These data indicate that Pg are present within autophagosomes and may use components of the autophagic pathway as a means to survive intracellularly. However, Pg presence within autophagosomes in KB cells could not be observed or detected. It is therefore likely that Pg uses different invasive mechanisms for different host cells. This and the role of HagA in invasion is currently being investigated further.

Animals↗

Replication, integration, and packaging of plasmid DNA following cotransfection with baculovirus viral DNA.

Infection-dependent replication assays have been used to identify numerous putative origins of baculovirus replication. However, plasmid DNA, when cotransfected into insect cells with Autographa californica multinucleocapsid nucleopolyhedrovirus (AcMNPV) DNA, replicates independently of any viral sequence in cis (11). Cotransfection of transfer plasmids and baculovirus DNA is a common procedure used in generating recombinant viruses and in measuring the level of gene expression in transient-expression assays. We have examined the fate of a series of vector plasmids in cotransfection experiments. The data reveal that these plasmids replicate following cotransfection and the replication of plasmid DNA is not due to acquisition of viral putative origin sequences. The conformation of plasmid DNA replicating in the cotransfected cells was analyzed and found to exist as high-molecular-weight concatemers. Ten to 25% of the replicated plasmid DNA was integrated into multiple locations on the viral genome and was present in progeny virions following serial passage. Sequence analysis of plasmid-viral DNA junction sites revealed no homologous or conserved sequences in the proximity of the integration sites, suggesting that nonhomologous recombination was involved during the integration process. These data suggest that while a rolling-circle mechanism could be used for baculovirus DNA replication, recombination may also be involved in this process. Plasmid integration may generate large deletions of the viral genome, suggesting that the process of DNA replication in baculovirus may be prone to generation of defective genomes.

Animals↗

Identification of CHIP, a novel tetratricopeptide repeat-containing protein that interacts with heat shock proteins and negatively regulates chaperone functions.

The chaperone function of the mammalian 70-kDa heat shock proteins Hsc70 and Hsp70 is modulated by physical interactions with four previously identified chaperone cofactors: Hsp40, BAG-1, the Hsc70-interacting protein Hip, and the Hsc70-Hsp90-organizing protein Hop. Hip and Hop interact with Hsc70 via a tetratricopeptide repeat domain. In a search for additional tetratricopeptide repeat-containing proteins, we have identified a novel 35-kDa cytoplasmic protein, carboxyl terminus of Hsc70-interacting protein (CHIP). CHIP is highly expressed in adult striated muscle in vivo and is expressed broadly in vitro in tissue culture. Hsc70 and Hsp70 were identified as potential interaction partners for this protein in a yeast two-hybrid screen. In vitro binding assays demonstrated direct interactions between CHIP and both Hsc70 and Hsp70, and complexes containing CHIP and Hsc70 were identified in immunoprecipitates of human skeletal muscle cells in vivo. Using glutathione S-transferase fusions, we found that CHIP interacted with the carboxy-terminal residues 540 to 650 of Hsc70, whereas Hsc70 interacted with the amino-terminal residues 1 to 197 (containing the tetratricopeptide domain and an adjacent charged domain) of CHIP. Recombinant CHIP inhibited Hsp40-stimulated ATPase activity of Hsc70 and Hsp70, suggesting that CHIP blocks the forward reaction of the Hsc70-Hsp70 substrate-binding cycle. Consistent with this observation, both luciferase refolding and substrate binding in the presence of Hsp40 and Hsp70 were inhibited by CHIP. Taken together, these results indicate that CHIP decreases net ATPase activity and reduces chaperone efficiency, and they implicate CHIP in the negative regulation of the forward reaction of the Hsc70-Hsp70 substrate-binding cycle.

Adenosine Triphosphatases↗

Oxidation of the skeletal muscle Ca2+ release channel alters calmodulin binding.

This study presents evidence for a close relationship between the oxidation state of the skeletal muscle Ca2+ release channel (RyR1) and its ability to bind calmodulin (CaM). CaM enhances the activity of RyR1 in low Ca2+ and inhibits its activity in high Ca2+. Oxidation, which activates the channel, blocks the binding of 125I-labeled CaM at both micromolar and nanomolar Ca2+ concentrations. Conversely, bound CaM slows oxidation-induced cross-linking between subunits of the RyR1 tetramer. Alkylation of hyperreactive sulfhydryls (<3% of the total sulfhydryls) on RyR1 with N-ethylmaleimide completely blocks oxidant-induced intersubunit cross-linking and inhibits Ca2+-free 125I-CaM but not Ca2+/125I-CaM binding. These studies suggest that 1) the sites on RyR1 for binding apocalmodulin have features distinct from those of the Ca2+/CaM site, 2) oxidation may alter the activity of RyR1 in part by altering its interaction with CaM, and 3) CaM may protect RyR1 from oxidative modifications during periods of oxidative stress.

Alkylation↗

Apolipoprotein A-I gene expression is regulated by cellular zinc status in hep G2 cells.

The influence of Zn on the expression of the apolipoprotein A-I (apoA-I) gene in Hep G2 cells was examined. Zn depletion was achieved with a low-Zn (ZD) medium prepared from Zn-free growth medium (Opti), a ZD medium containing Chelex 100-extracted fetal bovine serum (CHE), and a medium containing chelator 1, 10-phenanthroline (OP). Compared with those for their respective controls, cellular Zn levels were reduced by 55, 48, and 46% and apoA-I mRNA abundances were reduced by 20, 29, and 28% in Opti, CHE, and OP systems, respectively, after one passage in ZD media or 24 h in OP medium. To establish the specificity of Zn treatment, groups of ZD cells were treated with their respective control media for the last 24 h (ZDA) or normal cells were cultured with OP medium supplemented with Zn (OP-Zn). ZDA treatments partially normalized cellular Zn levels in the Opti system and restored or elevated apoA-I mRNA levels in the Opti or CHE system, respectively. Similarly, the OP-Zn treatment restored the cellular Zn and apoA-I mRNA levels. Furthermore, one passage of culture with Zn-supplemented media in both the Opti and CHE systems resulted in higher cellular Zn and apoA-I mRNA levels than those for controls. Most significantly, short-term high-Zn induction to normal cells markedly elevated the cellular Zn (3-fold) and apoA-I mRNA (5-fold) levels. Data derived from this study strongly suggest that the expression of apoA-I is regulated by cellular Zn status.

Apolipoprotein A-I↗

CaM kinase augments cardiac L-type Ca2+ current: a cellular mechanism for long Q-T arrhythmias.

Early afterdepolarizations (EAD) caused by L-type Ca2+ current (ICa, L) are thought to initiate long Q-T arrhythmias, but the role of intracellular Ca2+ in these arrhythmias is controversial. Rabbit ventricular myocytes were stimulated with a prolonged EAD-containing action potential-clamp waveform to investigate the role of Ca2+/calmodulin-dependent protein kinase II (CaM kinase) in ICa,L during repolarization. ICa,L was initially augmented, and augmentation was dependent on Ca2+ from the sarcoplasmic reticulum because the augmentation was prevented by ryanodine or thapsigargin. ICa,L augmentation was also dependent on CaM kinase, because it was prevented by dialysis with the inhibitor peptide AC3-I and reconstituted by exogenous constitutively active CaM kinase when Ba2+ was substituted for bath Ca2+. Ultrastructural studies confirmed that endogenous CaM kinase, L-type Ca2+ channels, and ryanodine receptors colocalized near T tubules. EAD induction was significantly reduced in current-clamped cells dialyzed with AC3-I (4/15) compared with cells dialyzed with an inactive control peptide (11/15, P = 0.013). These findings support the hypothesis that EADs are facilitated by CaM kinase.

Animals↗

Plasma insulin-like growth factor-I and serum IGF-binding protein 3 can be associated with the progression of breast cancer, and predict the risk of recurrence and the probability of survival in African-American and Hispanic women.

In vitro studies have shown that insulin-like growth factor (IGF) is a mitogen for breast cancer cells. However, the associations of plasma IGF-I with tumor histopathology in high-risk groups need further investigation. We hypothesize that plasma IGF-I and serum IGFBP3 concentrations in breast cancer patients may provide useful information on the progression of their disease, and determine the probability of recurrence and survival. We have carried out a retrospective study on 130 minority breast cancer patients. Plasma IGF-I and serum IGFBP3 were correlated with tumor histopathology, menopausal status, treatment modality, recurrence rates, and probability of survival. Plasma IGF-I and serum IGFBP3 were measured by radioimmunoassay. Our studies show that breast cancer patients have elevated plasma IGF-I and serum IGFBP3 levels. In addition we observed the following: IGF-I did not correlate with age and nodal stage. IGF-I and IGFBP3 increased with tumor size (T4). IGF-I did not correlate with estrogen receptor status, but did increase in progesterone-receptor-positive patients. IGF-I levels were higher in premenopausal patients and in women with cancer recurrence. Tamoxifen reduced IGF-I levels significantly and reduced the risk of recurrence. The survival probability was greater in patients with plasma IGF-I levels <120 ng/ml. In conclusion, lowering of plasma IGF-I may offer the following benefits: (a) reduce the risk of developing breast cancer in high-risk groups; (b) slow the progression of breast cancer in patients at early stages of cancer; (c) lower the risk of recurrence, and (d) increase the probability of survival.

Adult↗

Effects of inhaled nitric oxide and high-frequency ventilation in rabbits with meconium aspiration.

AIM: To evaluate effects of inhaled nitric oxide (iNO) in experimental meconium aspiration treated with high-frequency (HFV) or conventional mechanical ventilation (CMV). Ventilated adult rabbits had meconium instilled intratracheally resulting in respiratory failure as evidenced by more than 50% reduction of dynamic lung compliance (Cdyn) and increase in mean oxygenation index (OI) from 1 to 16. The animals were then allocated to 2 groups treated without (control) or with iNO at 20 ppm (NO). In each group the animals were initially ventilated with CMV or HFV mode for 3 h and then in a crossover fashion with HFV or CMV for another 3 h (CMV-->HFV, HFV-->CMV), respectively. In the first 3 h of treatment, the animals subjected to HFV-CMV in the control, and those with both HFV-CMV and CMV-HFV in the NO group had significantly reduced OI. In the subsequent 3 h, the animals in the control group with CMV-HFV did not improve in OI and those with HFV-CMV had deteriorated. In the NO group with both CMV-HFV and HFV-CMV moderate improvement of OI was observed. Platelet aggregation capability and counts were significantly decreased and bleeding time prolonged in animals receiving iNO treatment. These results suggest that both HFV alone and a combined treatment of iNO with either CMV or HFV are more effective in improving blood oxygenation than that of CMV in this animal model. The influence of iNO on platelet aggregation should be considered.

Administration, Inhalation↗

A transgenic rabbit model for human hypertrophic cardiomyopathy.

Certain mutations in genes for sarcomeric proteins cause hypertrophic cardiomyopathy (HCM). We have developed a transgenic rabbit model for HCM caused by a common point mutation in the beta-myosin heavy chain (MyHC) gene, R400Q. Wild-type and mutant human beta-MyHC cDNAs were cloned 3' to a 7-kb murine beta-MyHC promoter. We injected purified transgenes into fertilized zygotes to generate two lines each of the wild-type and mutant transgenic rabbits. Expression of transgene mRNA and protein were confirmed by Northern blotting and 2-dimensional gel electrophoresis followed by immunoblotting, respectively. Animals carrying the mutant transgene showed substantial myocyte disarray and a 3-fold increase in interstitial collagen expression in their myocardia. Mean septal thicknesses were comparable between rabbits carrying the wild type transgene and their nontransgenic littermates (NLMs) but were significantly increased in the mutant transgenic animals. Posterior wall thickness and left ventricular mass were also increased, but dimensions and systolic function were normal. Premature death was more common in mutant than in wild-type transgenic rabbits or in NLMs. Thus, cardiac expression of beta-MyHC-Q(403) in transgenic rabbits induced hypertrophy, myocyte and myofibrillar disarray, interstitial fibrosis, and premature death, phenotypes observed in humans patients with HCM due to beta-MyHC-Q(403).

Animals↗

Prognostic value of plasma HER-2/neu in African American and Hispanic women with breast cancer.

We examined the significance of plasma HER-2/neu as a clinical or biological marker for assessing the progression of breast cancer in African American and Hispanic women with similar socioeconomic status, similar health insurance, and similar access to health care delivery. Base line studies show the following: average age of our breast cancer patients was 48 for Hispanic and 53 for African American women. Most of our patients presented invasive ductal carcinoma, and there was no ethnic difference. A larger number of Hispanic women had stage III/IV disease at the time of diagnosis. There was no significant difference in the number of African American or Hispanic patients with ER positive or negative receptors. However, a larger number of Hispanic women had PR positive tumors, and a larger number of African American women had PR negative tumors. In general, there was no difference in the levels of HER-2/neu between the two ethnic groups. Patients with tumors >5 cm had elevated plasma HER-2/neu. However, there was no ethnic difference between tumor size and HER-2/neu levels. In addition, regional node status had no impact on plasma HER-2/neu. Patients with stage III/IV tumors had elevated plasma HER-2/neu. No ethnic difference was observed at either stage I/II or III/IV. ER positive or negative status had no significant impact on plasma HER-2/neu in either ethnic group. In contrast, PR positive patients showed elevated plasma HER-2/neu. Plasma HER-2/neu (>60 U/ml) was the strongest predictor of overall survival, visceral site metastasis, and local recurrence.

Adult↗

[Study of NAIP gene in spinal muscular atrophy].

OBJECTIVE: Investigating the correlation between the genotype and spinal muscular atrophy(SMA) clinical phenotype. METHODS: Neuronal apoptosis inhibitory protein(NAIP) gene analysis was performed by PCR amplification of exon 5 in 13 unrelated SMA patients with deletions of SMN gene. RESULTS: Two Type-I patients(2 M5, 40%) lacked the NAIP exon 5. CONCLUSION: The results suggest a possible correlation between NAIP gene deletion and the disease severity of SMA.

Child↗

Synergism between interleukin-6 and interleukin-1beta in hypothalamic serotonin release: a reverse in vivo microdialysis study in F344 rats.

The effects of interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) perfused locally into the anterior hypothalamus (AHY) on serotonin (5-hydroxytryptamine, 5-HT) release were investigated in the same region using in vivo microdialysis in conscious, freely moving F344 rats. IL-1beta (1 ng/rat) or IL-6 (50 ng/rat) injected directly into the AHY elicited a rapid and transient statistically significant increase in extracellular 5-HT levels (to 161% and 145% of the respective AUC (area under the curve) basal value, 100%). Intra-hypothalamic infusion of IL-1-receptor antagonist IL-1Ra (2 mug/rat) prevented this effect of IL-1beta, but not that of IL-6, suggesting an IL-1beta-independent mechanism for hypothalamic 5-HT release by this latter cytokine. Furthermore, intra-hypothalamic co-perfusion of IL-6 with IL-1beta at sub-optimal doses (10 ng/rat and 0.5 ng/rat, respectively) synergized in releasing hypothalamic 5-HT, thus providing in vivo evidence that both cytokines, IL-6 and IL-1beta are able to modulate the neuronal 5-HT response in the rat AHY.

Animals↗

[The mutation of deletion for glutathione S-transferase M1 gene in the tissue of hepatocellular carcinoma].

OBJECTIVE: To Study whether the mutation of deletion for glutathione S-transferase M1 (GSTM1) gene occurred during the development of hepatocellular carcinoma (HCC). METHODS: The genotypes of GSTM1 of 46 pairs of HCC tissue and the noncancerous liver tissue were detected by polymerase chain reaction (PCR). RESULTS: The frequency of GSTM1 null genotype for HCC tissue was 78.26%, but 65.22% for the noncancerous liver tissue (P<0.05). The GSTM1 null genotypes of 6 HCC tissue were transformed from the non- null genotypes of the noncancerous liver tissue. According to Hardy-Weinberg law., the rate of deletion mutation for GSTM1 gene was inferred to be 38.89%. CONCLUSION: The results suggested that the mutation of deletion for GSTM1 gene had occurred during the development of HCC.

Carcinoma, Hepatocellular↗

An experimental study of electro-acupuncture on auditory impairment caused by kanamycin in guinea pigs.

Frequency following response (FFR) and auditory brain stem evoked potential response (ABR) were used to determine the auditory acuity in evaluating the effect of electro-acupuncture treatment of kanamycin-induced auditory impairment in guinea pigs. The succinate dehydrogenase (SDH) activity and morphological changes of the inner ear receptors were examined under the light and scanning electron microscope in cochlear spread preparations. The results showed that 1) electro-acupuncture was effective but no significant differences were found among the stimulating wave forms; 2) Tinggong (SI 19), Yifeng (SJ 17), Shenshu (UB 23), Sanyinjiao (Sp 6), Zhubin (K 9) and Waiguan (SJ 5) are all effective acupoints, especially the combination of Tinggong (SI 19), Sanyinjiao (Sp 6) and Zhubin (K 9) acupoints; 3) improvement in the cochlear function and excitability of the cortical and lower auditory center and increase of the mitochondrial SDH activity and energy supply in hair cells might contribute to the mechanism of the treatment.

Animals↗

[Surveillance of hemorrhagic fever with renal syndrome and studies on its predictive indices in Jiangsu Province].

OBJECTIVE: To study its predictive significance of the direct use of surveillance data of hemorrhagic fever with renal syndrome (HFRS) collected in Jiangsu province, and find an indicator of choice for qualitative prediction. METHODS: A correlation analysis between the HFRS incidence rate and surveillance data collected in the past 12 years since 1986 was applied. RESULTS: There was significant relationship between HFRS incidence rate and indoor density of rattus norvegicus (Rn), proportions of mixed species of rats and Rn with HFRS virus, indices of mixed species of rats and Rn with virus in the spring Relationship between HFRS incidence rate and densities of outdoor mixed species of rats and Apodemus agrarius (Aa), and proportions of mixed species and Aa with virus was all significant in autumn and winter. And, there was significant relationship between HFRS incidence rate during the whole year and annual average density of rats, proportion of rats with virus, index of rats with virus. There was no significant relationship between HFRS incidence rate and human inapparent infection rate. The coefficients of correlation between HFRS incidence rate in the spring and indices of indoor mixed species of rats and Rn with virus were 0.8637 and 0.8295, respectively (P < 0.001). And, those between HFRS incidence rate and indices of outdoor mixed species of rats and Aa were 0.7089 and 0. 7258 in the autumn and winter, respectively (P < 0.01). The coefficients of correlation between HFRS incidence rate in the next spring and indices of outdoor mixed species and Aa with virus in the autumn were 0.7118 and 0.7113, respectively (P < 0.01). The coefficient of correlation between HFRS incidence rate during the whole year and annual average index of rats with virus was 0.9207 (P < 0.001). CONCLUSION: The index of rats with virus was an indicator of choice for qualitative prediction of HFRS, and the density of rats may be the secondary choice.

Animals↗

Site-directed mutagenesis analysis of draTG genes and their downstream region from Azospirillum brasilense Yu62.

draT and draG genes are involved in posttranslational regulation of nitrogenase activity of Azospirillum brasilense Yu62. Both genes and their downstream region were mutagenized by Kmr cassette insertions. Analysis of mutations introduced into the draTG region on the A. brasilense Yu62 chromosome showed that mutants affected in draT were incapable of regulating nitrogenase activity in response to ammonium. In contrast, a mutant with an insertion in draG was still capable of ADP-ribosylating dinitrogenase reductase in response to ammonium but was no longer able to recover activity after ammonium depletion. Analysis of mutations introduced into the draTG downstream region (the mutagenized site is about 2 kb downstream from draG) showed that the mutant had higher nitrogenase activity than the wild strain while growing in nitrogen-free medium and medium with 2 mmol/L ammonium. These results reveal that there is no gene required for nitrogen fixation in this mutagenized region, but it is possible that there are genes which play a role in regulating nitrogen fixation. The results of monitoring the expression of transcriptional nifH-lacZ gene fusion in the mutant YZ4 showed that the transcriptional regulation of nif gene in the mutant YZ4 was the same as that in the wild type.

Azospirillum brasilense↗

C677T mutation of the methylenetetrahydrofolate reductase gene among the Korean infants in Seoul city.

The missense mutation, C677T (Ala--->Val), in the methylenetetrahydrofolate reductase (MTHFR) gene, is related to hyperhomocysteinemia and is regarded as a risk factor for coronary artery disease and neural tube defects. The prevalence of this mutation was reported to differ among various ethnic groups, but there are few reports concerning Asian populations. We have investigated the frequencies of C677T mutation in 124 Korean infants (residents in Seoul city, Korea) and 115 Japanese adults (residents in Kobe city, Japan), and compared them with the reported data from other ethnic groups. The frequencies of the three genotypes in Koreans were as follows: C/C (wild homozygosity) 0.27, C/T (heterozygosity) 0.66, T/T (mutated homozygosity) 0.07, while those in Japanese were as follows: C/C 0.44, C/T 0.40, T/T 0.16. There was a marked difference in the genotype frequencies between the two populations (chi-square = 16.67, P = 0.0002), even though they are closely related in genetic background. The high C/T genotype frequency led to significant deviation from Hardy-Weinberg equilibrium (chi-square = 17.35, P = 0.00003). Deviation from Hardy-Weinberg equilibrium has not been found in any other ethnic groups. The high frequency of C/T genotype may offer Koreans a selective advantage.

Adult↗