Biomedical subjects

L Chen

Publications and source records attributed to L Chen.

At least 379 records · Page 21Linked to original sources

PubMed2000

Chemical synthesis and receptor binding of catfish somatostatin: a disulfide-bridged beta-D-Galp-(1-->3)-alpha-D-GalpNAc O-glycopeptide.

The glycopeptide hormone catfish somatostatin (somatostatin-22) has the amino acid sequence H-Asp-Asn-Thr-Val-Thr-Ser-Lys-Pro-Leu-Asn-Cys-Met-Asn-Tyr-Phe-Trp-Lys-Se r-Arg-Thr-Ala-Cys-OH; it includes a cyclic disulfide connecting the two Cys residues, and the major naturally occurring glycoform contains D-GalNAc and D-Gal O-glycosidically linked to Thr5. The linear sequence was assembled smoothly starting with an Fmoc-Cys(Trt)-PAC-PEG-PS support, using stepwise Fmoc solid-phase chemistry. In addition to the nonglycosylated peptide, two glycosylated forms of somatostatin-22 were accessed by incorporating as building blocks, respectively, Nalpha-Fmoc-Thr(Ac3-alpha-D-GalNAc)-OH and Nalpha-Fmoc-Thr(Ac4-beta-D-Gal-(1-->3)-Ac2-alpha-D-GalNAc)-O H. Acidolytic deprotection/cleavage of these peptidyl-resins with trifluoroacetic acid/scavenger cocktails gave the corresponding acetyl-protected glycopeptides with free sulfhydryl functions. Deacetylation, by methanolysis in the presence of catalytic sodium methoxide, was followed by mild oxidation at pH 7, mediated by Nalpha-dithiasuccinoyl (Dts)-glycine, to provide the desired monomeric cyclic disulfides. The purified peptides were tested for binding affinities to a panel of cloned human somatostatin receptor subtypes; in several cases, presence of the disaccharide moiety resulted in 2-fold tighter binding.

Amino Acid Sequence↗

PubMed2000

Learning- and cerebellum-dependent neuronal activity in the lateral pontine nucleus.

The effects of inactivation of cerebellar deep nuclei and the lateral pontine nucleus on classical eyeblink conditioning with tone or lateral reticular nucleus (LRN) stimulation as conditioned stimuli (CSs) were examined. Inactivation of cerebellar deep nuclei abolished eyeblink conditioned responses (CRs) when the CS was either a tone or LRN stimulation. Inactivation of the lateral pontine nucleus prevented only the acquisition and retention of tone-evoked eyeblink CRs. Multiple-unit recording demonstrated that when LRN stimulation was used as the CS, inactivation of the interpositus nucleus abolished learning-related neuronal activity in the lateral pontine nucleus, whereas inactivation of pontine nucleus had little effect on similar activity in the interpositus nucleus. Thus, the learning-induced neuronal activity in the lateral pontine nucleus was most likely driven by the cerebellar interpositus nucleus.

Acoustic Stimulation↗

PubMed2000

The DNA repair protein rad23 is a negative regulator of multi-ubiquitin chain assembly.

Rad23 is a nucleotide-excision repair protein with a previously unknown biochemical function. We determined that yeast and human Rad23 inhibited multi-ubiquitin (Ub) chain formation and the degradation of proteolytic substrates. Significantly, Rad23 could be co-precipitated with a substrate that contained a short multi-Ub chain. The UV sensitivity of rad23Delta was reduced in mutants lacking the E2 enzyme Ubc4, or the multi-Ub chain-promoting factor Ufd2. These studies suggest that the stability of proteolytic substrates is governed by the competing action of multi-Ub chain-promoting and chain-inhibiting factors. The stabilization of DNA repair and stress factors could represent an important biological function of Rad23.

DNA Repair↗

PubMed2000

Modulation of T-cell-mediated immunity in tumor and graft-versus-host disease models through the LIGHT co-stimulatory pathway.

LIGHT was recently described as a member of the tumor necrosis factor (TNF) 'superfamily'. We have isolated a mouse homolog of human LIGHT and investigated its immunoregulatory functions in vitro and in vivo. LIGHT has potent, CD28-independent co-stimulatory activity leading to T-cell growth and secretion of gamma interferon and granulocyte-macrophage colony-stimulating factor. Gene transfer of LIGHT induced an antigen-specific cytolytic T-cell response and therapeutic immunity against established mouse P815 tumor. In contrast, blockade of LIGHT by administration of soluble receptor or antibody led to decreased cell-mediated immunity and ameliorated graft-versus-host disease. Our studies identify a previously unknown T-cell co-stimulatory pathway as a potential therapeutic target.

Amino Acid Sequence↗

PubMed2000

MJ0109 is an enzyme that is both an inositol monophosphatase and the 'missing' archaeal fructose-1,6-bisphosphatase.

In sequenced genomes, protein coding regions with unassigned function constitute between 10 and 50% of all open reading frames. Often key enzymes cannot be identified using sequence homology searches. For example, despite the fact that methanogens have an apparently functional gluconeogenesis pathway, standard tools have been unable to identify a fructose-1,6-bisphosphatase (FBPase) gene in the sequenced Methanoccocus jannaschii genome. Using a combination of functional and structural tools, we have shown that the protein product of the M. jannaschii gene MJ0109, which had been tentatively annotated as an inositol monophosphatase (IMPase), has both IMPase and FBPase activities. Moreover, several gene products annotated as IMPases from different thermophilic organisms also possess FBPase activity. Thus, we have found the FBPase that was 'missing' in thermophiles and shown that it also functions as an IMPase.

Amino Acid Sequence↗

PubMed2000

T-cell killing of heterogenous tumor or viral targets with bispecific chimeric immune receptors.

We have previously described several novel chimeric immune receptors (CIRs) that redirect human T cells to kill malignant or HIV-infected cells. These CIRs comprise a cancer- or virus-specific ligand or single-chain antibody fused to the signaling domain of the T-cell receptor CD3-zeta subunit. Binding of the ligand- or antibody-based CIR to the target antigen (Ag) triggers T-cell-mediated cytolysis of the tumor- or virus-infected cell independent of target cell major histocompatibility complex class I expression. A new type of CIR was developed to mediate the lysis of cells that expressed one or more distinct viral or tumor Ags; three bispecific CIRs (BCIRs) were generated that recognized the carcinoembryonic Ag (CEA) and TAG-72 tumor Ags or, alternatively, distinct epitopes in the HIV envelope (HIVenv). T cells expressing the antitumoral Ag BCIR lysed both CEA- and TAG-72-expressing targets and did not kill Ag-negative targets or target cells expressing other members of the CEA family. Similarly, T cells expressing the anti-HIVenv BCIR lysed target cells expressing both the wild-type HIVenv and a mutant HIVenv that lacked the epitopes recognized by the monospecific CIRs. This approach permits the generation of T cells with a broader spectrum of activity capable of killing virus-infected cells and malignant cells and reduces the potential of progression of disease due to Ag loss variants.

Amino Acid Sequence↗

PubMed2000

One-step electropolymeric co-immobilization of glucose oxidase and heparin for amperometric biosensing of glucose.

Electropolymerization was used for the co-deposition of glucose oxidase and heparin onto metal electrode transducers. Such electropolymeric co-entrapment within a non-conducting poly(1,2-phenylenediamine) (PPD) film imparts both biocatalytic and anticoagulation activities onto the transducer, and greatly improves the performance of the sensor after exposure to whole blood. Essentially identical glucose signals are observed before and after exposure to blood samples. Scanning electron micrographs after such exposure reveal no platelet deposition or formation of a fibrin "clot". The effect of the heparin co-immobilization on the glucose response is examined. Improved biocompatibility is reported also in connection with a needle-type carbon paste biosensor configuration. The simultaneous localization of the enzyme and heparin offers great promise for simplifying the preparation of enzyme electrodes and designing biocompatible implantable glucose biosensors.

Biosensing Techniques↗

PubMed2000

Neutrophils play a critical role in the pathogenesis of experimental cerebral malaria.

The role of neutrophils in experimental cerebral malaria (ECM) is not well understood. In this study we used a MoAb, RB6-8C5, to deplete the peripheral neutrophils of ECM-susceptible CBA/NSlc mice 24 h before Plasmodium berghei ANKA (PbA) infection. We found that early neutrophil depletion prevented the development of ECM and dramatically decreased the sequestration of monocytes and microhaemorrhage in the brain. The depletion of neutrophils also down-regulated tumour necrosis factor-alpha, interferon-gamma and IL-2 mRNAs and abrogated IL-12p40 mRNA expression in the brain as examined by competitive reverse transcriptase-polymerase chain reaction. Although depletion of neutrophils decreased the expression of Th1 cytokines in both spleen and brain, our results did not show the shift of a Th1 to a Th2 immune response since there was no obvious augmentation of expression of Th2 cytokine mRNAs (IL-4 and IL-10). We conclude that neutrophils play a role in the pathogenesis of ECM via enhancement of the expression of Th1 cytokines in the brain.

Animals↗

PubMed2000

Classification of impairment of shoulder abduction in obstetric brachial plexus palsy and its clinical significance.

Shoulder abduction was studied in 25 cases of obstetric brachial plexus palsy (OBPP). According to muscle function, electromyographic features and X-ray examination, impairment of shoulder abduction could be classified into dynamic, resistant or combined types. Five cases were the dynamic type, which was characterized by paralysis of the shoulder abductors. Fifteen cases were categorized as the resistant type, with contracture of the subscapularis muscle, co-contraction of latissimus dorsi and teres major muscles and secondary disorders of the shoulder joint. Five cases were classified as the combined type in which there were both dynamic and resistant factors. Appropriate management and surgical procedures in the shoulder affected by OBPP depend on the pathological classification.

Adolescent↗

PubMed2000

The beginning of a structural reform: reorganising the front line of a mental health service.

National and State priorities for mental health services have directed emphasis towards early intervention and prevention. One of the key priorities is to ensure that entry to mental health services is efficient, effective and accountable. This study describes the process of restructuring the front line of a large and complex mental health service. Adopting the total quality management approach, all stakeholders in the service collaboratively developed a single set of protocols and guidelines to achieve standardisation of documentation, assessment of risks and urgency, and to improve the overall quality of the service.

Australia↗

PubMed2000

Improving the utilisation of bilingual counsellors within a public sector mental health service.

An urban, public sector Area Mental Health Program reviewed its own bilingual counsellor program as part of an Area-wide quality improvement project, and found that the counsellors' roles needed to be better defined; that mainstream staff needed to have more access to their expertise as cultural consultants; and that their function as an Area team, rather than as service based staff, needed to be encouraged. The bilingual counsellors decided to take up this challenge, and with the support of the Area Director of Mental Health, worked together to redefine their roles.

Communication Barriers↗

PubMed2000

Suppression of transcription factor NF-kappaB activity by Bcl-2 protein in NIH3T3 cells: implication of a novel NF-kappaB p50-Bcl-2 complex for the anti-apoptotic function of Bcl-2.

Bcl-2 can suppress apoptosis by controlling genes that encode proteins required for programmed cell death and by interference with peroxidative damage. Overexpression of Bcl-2 in NIH3T3 cells can prevent GSNO-induced (S-nitrosoglutathione-induced) apoptosis. The experimental results indicated that activation of NF-kappaB by GSNO is involved in inducing apoptosis. Surprisingly, we found that Bcl-2 delayed the release of IkB by formation of a Bcl-2-NF-kappaB complex (p50-p65-IkappaB) in the cytoplasm during cell apoptosis. Furthermore, a novel Bcl-2-p50 complex was found in the nucleus. These features were only observed in Bcl-2-transfected cells but not in the parental NIH3T3 cells. Overexpression of Bcl-2 suppressed the levels of c-myc, a target gene of NF-kappaB, and influenced the DNA-binding activity of NF-kappaB during GSNOinduced apoptosis. We suggest that the Bcl-2-p50 complex inhibits NF-kappaB DNA-binding activity by competing with the p65-p50 heterodimer for the DNA-binding site in the nucleus. Finally, it has been demonstrated that the anti-apoptotic potential of Bcl-2 may be attributed to its complexing with p50 in the nucleus that leads to blockage of nuclear gene expression.

3T3 Cells↗

PubMed2000

Extracellular adenosine 5'-ATP-induced calcium signaling in isolated vestibular ganglion cells of the guinea pig.

Extracellular adenosine 5'-triphosphate (ATP)-induced intracellular calcium concentration ([Ca2+]i) changes in acutely isolated vestibular ganglion cells (VGCs) of the guinea pig were investigated using the Ca2+ -sensitive dye Fura-2. Extracellular ATP induced an increase in [Ca2+]i in VGCs in a dose-dependent manner. ATP induced an increase in [Ca2+]i even in the absence of extracellular Ca2+ (1 mM Ethylene Glycol-bis (beta-aminoethyl Ether) N,N,N',N'-Tetraacetic Acid (EGTA)), thus suggesting that ATP induces Ca2+ release from the intracellular stores. The P2-receptor antagonists suramin and reactive blue 2 inhibited the ATP-induced [Ca2+]i increase in a dose-dependent manner. The P1-receptor agonist adenosine did not induce any changes in [Ca2+]i. These results suggest that VGCs may possess a P2-purinergic receptor but not a P1-purinergic receptor. La3+, a receptor-mediated calcium channel blocker, inhibited the ATP-induced [Ca2+]i increase but, in contrast, nifedipine, a L-type calcium channel blocker, did not. These results suggest that ATP induces both a Ca2+ -release from the intracellular stores and a Ca2+ influx from the extracellular space through La3+ -sensitive and nifedipine-insensitive Ca2+ channels in VGCs. Our results also suggest that extracellular ATP may act as a neurotransmitter or neuromodulator of the vestibular peripheral system in the guinea pig.

Adenosine Triphosphate↗

PubMed2000

Air particulate pollution and hospital admissions for chronic obstructive pulmonary disease in Reno, Nevada.

This study assessed the association between ambient PM(10) pollution and daily hospital admissions for chronic obstructive pulmonary disease (COPD) in Reno-Sparks, Nevada, for the period 1990-1994. All three hospitals in the region were included. There was a total of 3115 admissions for COPD during this period. Daily ambient PM(10) values were available from one of seven air monitoring stations in this region. Weather variables including temperature and wind speed were also collected from this station. The daily average concentration of PM(10) was 36.55 microg/m(3). The generalized additive model (GAM) was used in the whole analysis. After adjusting for the effects of weather variables, day of week, seasons, and time trend, the results show that PM(10) is a statistically significant predictor for daily hospital admissions for COPD. The relative risk (RR) of hospital admissions for COPD for an interquartile increase (26.6 microg/m(3)) of the 24-h average level of PM(10) is 1.049 (95% CI 1.011-1.087).

Air Pollutants↗

PubMed2000

Elementary school absenteeism and air pollution.

This study assessed the association between ambient air pollution and daily elementary school absenteeism in Washoe County, NV, between 1996 and 1998. All 57 elementary schools in Washoe County in northern Nevada were included in the data set. There was a total of 27,793 student enrollments during this study period. The daily average absence rate was 5.09% (+/-1.54%). Air pollutant values including PM(10), O(3), and CO were obtained from seven air monitoring stations. Weather variables were collected from five of seven stations and from the Western Regional Climate Center. The daily average concentrations of PM(10), CO, and O(3) were 32.44 microg/m(3), 2.73 ppm, and 37.45 ppb, respectively. Student absenteeism was regressed on the three air pollutants, weather variables, and other confounding factors, using autoregression analysis. After adjusting for the effects of weather variables, day of the week, month, and holiday indicators, and time trend, we found that CO and O(3) were statistically significant predictors of daily absenteeism in elementary schools. For every 1.0 ppm and 50 ppb increase in CO and O(3), the absence rate would increase 3.79% (95% CI 1.04-6.55%) and 13.01% (95% CI 3.41-22.61%), respectively. However, PM(10) values were negatively correlated with school absenteeism.

Absenteeism↗

PubMed2000

Induction of peroxiredoxins in transplanted livers and demonstration of their in vitro cytoprotection activity.

Peroxiredoxin (Prx)-I and -II belong to a new class of antioxidants. Here, we report that they are induced by ischemia/reperfusion (I/R) in transplanted livers. Hypothesizing that Prxs are induced to protect liver from oxidative damage, we transduced these human genes into murine NIH-3T3 cells. The overexpressed Prxs made the cells more resistant to t-butylhydroperoxide-induced apoptosis. These results indicate that Prx-I and Prx-II are induced by the transplantation process and can protect cells against oxidant damage in tissue culture. Thus, proper genetic manipulations of Prxs may be useful in increasing the success rate of organ transplantation.

3T3 Cells↗

PubMed2000

The roles of E6-AP and MDM2 in p53 regulation in human papillomavirus-positive cervical cancer cells.

The p53 tumor suppressor is regulated by the MDM2 oncoprotein through a negative feedback mechanism. MDM2 promotes the ubiquitination and proteasome-dependent degradation of p53, possibly by acting as a ubiquitin ligase. In cervical cancer cells containing high-risk human papillomaviruses (HPV), p53 is also targeted for degradation by the HPV E6 oncoprotein in combination with the cellular E6-AP ubiquitin ligase. In this report, we describe the identification of efficient antisense oligonucleotides against human E6-AP. The roles of MDM2 and E6-AP in p53 regulation were investigated using a novel E6-AP antisense oligonucleotide and a previously characterized MDM2 antisense oligonucleotide. In HPV16-positive and HPV-18 positive cervical cancer cells, inhibition of E6-AP, but not MDM2, expression results in significant induction of p53. In HPV-negative tumor cells, p53 is activated by inhibition of MDM2 but not E6-AP. Furthermore, treatment with both E6-AP and MDM2 antisense oligonucleotides in HPV-positive cells does not lead to further induction of p53 over inhibition of E6-AP alone. Therefore, E6-AP-mediated degradation is dominant over MDM2 in cervical cancer cells but does not have a significant role in HPV-negative cells.

Cell Survival↗

PubMed2000

The ycf 9 (orf 62) gene in the plant chloroplast genome encodes a hydrophobic protein of stromal thylakoid membranes.

There are still some open reading frames, orfs, with unknown function in the higher plant chloroplast genome. Of these conserved orfs, designated as ycfs (hypothetical chloroplast open reading frames), one is ycf 9 (orf 62) in the transcription unit with the psbC and psbD genes. The aim of this work was to investigate the function of ycf 9 by insertional inactivation of the gene with a selectable marker cassette, consisting of the aadA coding region connected to the trc promoter and rrnB terminator. This cassette was inserted 19 bp downstream from the start of the coding region of the tobacco ycf 9 gene. Two DNA constructs with the aadA cassette in opposite orientations were precipitated on 1 micron gold particles and delivered into leaves of Nicotiana tabacum, cultivar Samsun, by the biolistic method. Spectinomycin-resistant plants regenerated following bombardment with only the construct containing the aadA gene in the opposite orientation as ycf 9. In spite of several subsequent regeneration cycles on spectinomycin, the transplastomic plants did not reach homoplasmicity. This suggests that the ycf 9 gene product is essential for chloroplast function. Using a polyclonal antibody raised against the inner part of the gene product, the polypeptide was localized in the stromal thylakoid membranes of chloroplasts.

Amino Acid Sequence↗