Biomedical subjects

L Zheng

Publications and source records attributed to L Zheng.

At least 55 records · Page 3Linked to original sources

PubMed2001

When genetic distance matters: measuring genetic differentiation at microsatellite loci in whole-genome scans of recent and incipient mosquito species.

Genetic distance measurements are an important tool to differentiate field populations of disease vectors such as the mosquito vectors of malaria. Here, we have measured the genetic differentiation between Anopheles arabiensis and Anopheles gambiae, as well as between proposed emerging species of the latter taxon, in whole genome scans by using 23-25 microsatellite loci. In doing so, we have reviewed and evaluated the advantages and disadvantages of standard parameters of genetic distance, F(ST), R(ST), (delta mu)(2), and D. Further, we have introduced new parameters, D' and D(K), which have well defined statistical significance tests and complement the standard parameters to advantage. D' is a modification of D, whereas D(K) is a measure of covariance based on Pearson's correlation coefficient. We find that A. gambiae and A. arabiensis are closely related at most autosomal loci but appear to be distantly related on the basis of X-linked chromosomal loci within the chromosomal Xag inversion. The M and S molecular forms of A. gambiae are practically indistinguishable but differ significantly at two microsatellite loci from the proximal region of the X, outside the Xag inversion. At one of these loci, both M and S molecular forms differ significantly from A. arabiensis, but remarkably, at the other locus, A. arabiensis is indistinguishable from the M molecular form of A. gambiae. These data support the recent proposal of genetically differentiated M and S molecular forms of A. gambiae.

Animals↗

PubMed2001

Astrocytes, not microglia, are the main cells responsible for viral persistence in Theiler's murine encephalomyelitis virus infection leading to demyelination.

The BeAn strain of Theiler's murine encephalomyelitis virus (TMEV) persists in the CNS and produces a chronic inflammatory demyelinating disease that is an animal model for human multiple sclerosis (MS). The mechanisms leading to TMEV-induced demyelination are still under study but most likely involve both immune-mediated and virus induced damage to cells in the CNS, both depending on viral persistence. It is therefore important to identify the cells in which continued virus production is permitted. In this study, we looked at virus infection in primary astrocytes, microglia and oligodendrocytes, derived from brains of neonatal susceptible SJL/J mice. As evidenced by Western blots and immunocytochemistry, we were able to detect viral antigens in all these brain-derived cells. In addition, we extended the study to spinal cord tissues from mice suffering TMEV-induced disease. Immunohistochemistry staining with anti-TMEV sera and antibodies to specific cell markers detected viral antigens in all these cells. We then asked the question whether viral antigen present in these cells, particularly in microglia/macrophages, represented true viral replication or not. By using different techniques, including immunoprecipitation experiments and the very sensitive method of negative RNA detection through RNase protection assay, we show that both astrocytes and oligodendroglia permit de novo viral replication and viral protein synthesis but with only minimal cytopathic effects. Of these two cell types, astrocytes carry the brunt of viral replication. In microglia, on the other hand, viral replication is restricted since only minimal amounts of negative RNA copies can be demonstrated, while there are clear signs that some of these cells undergo apoptosis. These findings show that the main cell for viral replication is the astrocyte, rather than the microglia/macrophage. Most of the viral antigen present in macrophages, therefore, is probably the result of phagocytosis, rather than actual viral replication. In view of the demonstrated presence of viral replication in astrocytes and of great amounts of viral antigens in microglia/macrophages, it is possible that both types of cells act as antigen presenting cells during this immunopathological disease.

Animals↗

PubMed2001

Solid-phase analysis method for (S)-[18F]fluorocarazolol and its metabolites.

(S)-[18F]Fluorocarazolol is a radiopharmaceutical developed to quantitatively assess beta-adrenergic receptors in vivo via positron emission tomography imaging. Since radioactive metabolites of (S)-[18F]fluorocarazolol rapidly appear in the plasma, methods for conveniently and reliably evaluating plasma for (S)-[18F]fluorocarazolol content are required. Here we present methods and validation of an approach using commercial extraction cartridges that is faster and more convenient than an approach using internal-surface reverse-phase chromatography but yields comparable results.

Adrenergic beta-Antagonists↗

PubMed2001

BRCA1 mediates ligand-independent transcriptional repression of the estrogen receptor.

Mutational inactivation of BRCA1 confers a cumulative lifetime risk of breast and ovarian cancers. However, the underlying basis for the tissue-restricted tumor-suppressive properties of BRCA1 remains poorly defined. Here we show that BRCA1 mediates ligand-independent transcriptional repression of the estrogen receptor alpha (ERalpha), a principal determinant of the growth, differentiation, and normal functional status of breasts and ovaries. In Brca1-null mouse embryo fibroblasts and BRCA1-deficient human ovarian cancer cells, ERalpha exhibited ligand-independent transcriptional activity that was not observed in Brca1-proficient cells. Ectopic expression in Brca1-deficient cells of wild-type BRCA1, but not clinically validated BRCA1 missense mutants, restored ligand-independent repression of ERalpha in a manner dependent upon apparent histone deacetylase activity. In estrogen-dependent human breast cancer cells, chromatin immunoprecipitation analysis revealed the association of BRCA1 with ERalpha at endogenous estrogen-response elements before, but not after estrogen stimulation. Collectively, these results reveal BRCA1 to be a ligand-reversible barrier to transcriptional activation by unliganded promoter-bound ERalpha and suggest a possible mechanism by which functional inactivation of BRCA1 could promote tumorigenesis through inappropriate hormonal regulation of mammary and ovarian epithelial cell proliferation.

Adenocarcinoma↗

PubMed2001

Stimulation of eukaryotic flap endonuclease-1 activities by proliferating cell nuclear antigen (PCNA) is independent of its in vitro interaction via a consensus PCNA binding region.

Interaction between human flap endonuclease-1 (hFEN-1) and proliferating cell nuclear antigen (PCNA) represents a good model for interactions between multiple functional proteins involved in DNA metabolic pathways. A region of 9 conserved amino acid residues (residues Gln-337 through Lys-345) in the C terminus of human FEN-1 (hFEN-1) was shown to be responsible for the interaction with PCNA. Our current study indicates that 4 amino acid residues in hFEN-1 (Leu-340, Asp-341, Phe-343, and Phe-344) are critical for human PCNA (hPCNA) interaction. A conserved PCNA interaction motif in various proteins from assorted species has been defined as Q(1)X(2)X(3)(L/I)(4)X(5)X(6)F(7)(F/Y)(8), although our results fail to implicate Q(1) (Gln-337 in hFEN-1) as a crucial residue. Surprisingly, all hFEN-1 mutants, including L340A, D341A, F343A, and F344A, retained hPCNA-mediated stimulation of both exo- and flap endonuclease activities. Furthermore, our in vitro assay showed that hPCNA failed to bind to the scRad27 (yeast homolog of FEN-1) nuclease. However, its nuclease activities were significantly enhanced in the presence of hPCNA. Four additional Saccharomyces cerevisiae scRad27 mutants, including multiple alanine mutants and a deletion mutant of the entire PCNA binding region, were constructed to confirm this result. All of these mutants retained PCNA-driven nuclease activity stimulation. We therefore conclude that stimulation of eukaryotic hFEN-1 nuclease activities by PCNA is independent of its in vitro interaction via the PCNA binding region.

Amino Acid Motifs↗

PubMed2001

Cloning, expression patterns, and chromosome localization of three human and two mouse homologues of GABA(A) receptor-associated protein.

Type A receptors of gamma-aminobutyric acid (GABA), an inhibitory neurotransmitter, contain alpha, beta, delta, gamma, and rho subunits. The gamma subunit has four subtypes: gamma1, gamma2, gamma3, andgamma4. GABA(A) receptor-associated protein (GABARAP) was previously demonstrated to act as a linker protein between microtubules and the gamma2 subunit of GABA(A) receptors. However, no other linker proteins have been identified as mediating the linkage of microtubules and the remaining subunits of GABA(A) receptors. In this study we identified three human paralogues (GABARAPL1, GABARAPL2, and GABARAPL3) and two mouse orthologues (Gabarapl1 and Gabarapl2) of human GABARAP, all of which encoded 117 amino acids, as does Gabarapl. The expression patterns of GABARAPL1, GABARAPL2, and GABARAP in 16 adult tissues showed that they were expressed ubiquitously. The expression levels of GABARAPL1 as a 2.3-kb transcript were very high in brain, heart, peripheral blood leukocytes, liver, kidney, placenta, and skeletal muscle, very low in thymus and small intestine, and moderate in other tissues tested. The unique 1.35-kb transcript of GABARAPL2 was expressed at high levels in heart, brain, testis, prostate, ovary, spleen, and skeletal muscle, at very low levels in lung, thymus, and small intestine, and moderately in other tissues tested. For GABARAP, a 1.3-kb transcript was abundantly expressed in all tested tissues with small variation. The expression patterns of Gabarapl1 and Gabarapl2 were similar to those of their counterparts in human. In addition, GABARAPL1 was localized to human chromosome 12p12.3 and GABARAPL2 to 16q22.3-q24.1 by RH mapping, while GABARAP and GABARAPL3 were found to be localized at chromosomes 17p13.2 and 15q25.1, respectively, by searching the related databases. Sequence comparison of the cDNAs and their corresponding genomic sequences shows that GABARAP, GABARAPL1, and GABARAPL2 are composed of four exons each, while GABARAPL3 is distributed only at one exon.

Adaptor Proteins, Signal Transducing↗

PubMed2001

The death effector domain-associated factor plays distinct regulatory roles in the nucleus and cytoplasm.

Homophilic interactions of death effector domains (DEDs) are crucial for the signaling pathways of death receptor-mediated apoptosis. The machinery that regulates proper oligomerization and autoactivation of procaspase-8 and/or procaspase-10 during T lymphocyte activation determines whether the cells will undergo caspase-mediated apoptosis or proliferation. We screened a yeast two-hybrid library by using the DEDs contained in the prodomains of procaspase-8 and procaspase-10 and isolated a DED-associated factor (DEDAF) that interacts with several DED-containing proteins but does not itself contain a DED. DEDAF is highly conserved between human and mouse (98% amino acid identity) and is homologous to a nuclear regulatory protein YAF-2. DEDAF is expressed at the highest levels in lymphoid tissues and placenta. DEDAF interacts with FADD, procaspase-8, and procaspase-10 in the cytosol as well as with the DED-containing DNA-binding protein (DEDD) in the nucleus. At the cell membrane, DEDAF augmented the formation of CD95-FADD-caspase-8 complexes and enhanced death receptor- as well as DED-mediated apoptosis. In the nucleus, DEDAF caused the DEDD protein to relocalize from subnuclear structures to a diffuse distribution in the nucleoplasm. Our data therefore suggest that DEDAF may be involved in the regulation of both cytoplasmic and nuclear events of apoptosis.

Amino Acid Sequence↗

PubMed2001

The retinoblastoma gene: a prototypic and multifunctional tumor suppressor.

Genome instability has been implicated in the generation of multiple somatic mutations that underlie cancer. Germline mutation in the retinoblastoma (RB) gene leads to tumor formation in both human and experimental animal models, and reintroduction of wild-type RB is able to suppress neoplastic phenotypes. Rb governs the passage of cells through the G1 phase-restriction point and this control is lost in most cancer cells. Rb has also been shown to promote terminal differentiation and prevent cell cycle reentry. Recent studies implicate Rb in mitotic progression, faithful chromosome segregation, checkpoint control, and chromatin remodeling, suggesting that Rb may function in the maintenance of genome integrity. It is likely that Rb suppresses tumor formation by virtue of its multiple biological activities. A single protein capable of performing multiple antioncogenic functions may be a common characteristic of other tumor suppressors including p53 and BRCA1/2.

Animals↗

PubMed2001

MR imaging-guided radiofrequency thermal ablation in the porcine brain at 0.2 T.

The aim of this study was to test the hypotheses that (a) MR imaging-guided radiofrequency (RF) thermal ablation is safe and feasible in porcine brain using an open C-arm-shaped low-field MR system, and that (b) induced thermal lesion size can be predicted using low-field MR imaging. Magnetic resonance-guided RF ablation was performed in the cerebral frontal lobes of six pigs. An 18-G monopolar RF electrode was inserted into the porcine brain using MR image guidance and RF was then applied for 10 min. After post-procedure imaging (T2-weighted, T1-weighted before and after gadodiamide administration), the pigs were killed and the brains were used for pathologic examination. Successful RF electrode placement was accomplished in all cases without complications; total magnet time ranged from 73 to 189 min. The thermal lesion size varied from 10 to 12 mm perpendicular to the electrode track and was easily visualized on T2-weighted and enhanced T1-weighted images. Enhanced T1-weighted imaging demonstrated the highest brain-to-RF thermal lesion contrast-to-noise ratio with an average of 1.5 +/- 1.6. Enhanced T1-weighted imaging never underestimated pathologic lesion diameter with a mean difference of 2.3 +/- 1.0 mm and a radiologic/pathologic correlation of 0.69. Magnetic resonance imaging-guided RF thermal ablation is feasible and safe in the porcine brain using an open MR low-field system. Induced-thermal lesion size can best be monitored using enhanced T1-weighted images. In the future, RF ablation under low-field MR guidance may offer an alternative treatment option for primary and secondary brain tumors.

Animals↗

PubMed2001

Labelling and binding of poly-(L-lysine) to functionalised gold surfaces. Combined FT-IRRAS and XPS characterisation.

We compare herein the interfacial reactivity of self-assembled monolayers (SAMs) of 11-mercaptoundecanoic acid (MUA), 1-undecanethiol (UDT) and 11-mercaptoundecanol (MUD) on gold surfaces towards aqueous solutions of poly-(L-lysine) (PL). Liquid-phase labelling of PL with the alkyne dicobalt hexacarbonyl cluster 1 combined with analysis of the substrates by Fourier transform infrared reflection-absorption spectroscopy (FT-IRRAS) and X-ray photoelectron spectroscopy (XPS) revealed that irreversible binding of PL occurred in all cases. However, the mechanism of binding involved differed markedly from one monolayer to the other. The main mode of interaction of PL to MUA SAM was of electrostatic nature between the terminal carboxylate of MUA and the ammonium groups of PL. For a similar number of bound thiolate molecules, the UDT adsorbed layer was found less continuous than the MUA one, allowing a higher fraction of PL to directly bind to the gold surface. As for MUD, very little thiolate molecules were adsorbed, leaving bare gold surface areas for non specific adsorption of PL.

Journal Article↗

PubMed2001

Degumming of ramie fibers by alkalophilic bacteria and their polysaccharide-degrading enzymes.

Three strains of alkalophilic bacteria, Bacillus sp. NT-39, NT-53 and NT-76, were selected for the degumming of ramie fibers and production of polysaccharide-degrading enzymes. After 48 h of incubation with the strains, the loss of the gum might amount to 5.0% or more of the fibers and a number of polysaccharide-degrading enzymes were secreted to the culture supernatants. The residual gum of the fibers decreased to 9.4% after 5 h of enzymatic degumming. Analysis of gum contents and enzyme activities revealed that pectate lyase and xylanase played an important role in the degradation of residual gum. Enzymatic degumming resulted in an increment of 5.4 ISO units in fiber brightness, whereas the reduction in bundle breaking tenacity of the fibers was less than 5.%. The results confirmed that degumming of ramie fibers by alkalophilic bacteria and their enzymes had substantial advantages.

Bacillus↗

PubMed2001

Sexual behaviour and contraceptive use among unmarried, young women migrant workers in five cities in China.

This paper reports the results of exploratory research on reproductive and sexual health knowledge and sexual behaviour of young, unmarried women who migrate to cities from rural areas for work, and their access to and needs in relation to family planning in Beijing, Guangzhou, Shanghai, Guiyang and Taiyuan, in China. Focus group discussions were conducted with 146 young women aged 16-25 and 58 in-depth interviews with key informants. Some of the young female migrant workers were sexually active and living with their boyfriends, most of whom expected to marry each other. Most of the women lacked basic information about reproduction and contraception, and did not know where or how to obtain contraception. There were social, psychological and economic barriers to accessing services. Only a small proportion of those who were unmarried were using contraception, so induced abortion was often the outcome of unprotected premarital sex. Pleasing male partners also played an important role in unprotected sex. The training, attitudes and approach of the entire family planning service system in relation to unmarried and young people in China, including this migrant population, needs to be reorientated so as to provide them with appropriate and adequate services.

Abortion, Induced↗

PubMed2001

Iron overload and nitric oxide-derived oxidative stress following lung transplantation.

BACKGROUND: Reactive oxygen species (ROS) may contribute to airway injury and the development of the bronchiolitis obliterans syndrome (BOS) following lung transplantation (LT). Chemically active iron released from ferritin stores and nitric oxide (NO)-derived radicals may add to the oxidative burden. METHODS: We determined the concentrations of ferritin and the aqueous NO derivative nitrite (NO2(-)) within bronchoalveolar lavage fluid (BALF) of 14 stable LT recipients (ST) and 7 subjects with BOS and 21 normal controls. We also assessed the relationship between BALF ferritin and hemosiderin-laden macrophages (HLMs) using a hemosiderin score (HS) and determined BALF albumin concentration as a marker of microvascular leakage. RESULTS: BALF ferritin concentrations and HSs were significantly elevated in LT recipients overall compared with normal controls (p < 0.05). BALF NO2(-) levels were elevated in BOS subjects and STs compared with normal controls (p = 0.002 and p = 0.09, respectively), but there was no difference between transplant groups. BALF albumin concentrations were elevated in BOS patients compared with normal controls (p = 0.02) and ST (p = 0.05), but there was no difference between STs and controls. There was a significant relationship between BALF ferritin concentration and HS in LT recipients overall (r(s) = 0.7, p < 0.001). In BOS subjects, but not ST, BALF ferritin was significantly related to BALF albumin (r(s) = 0.8, p = 0.05) and there was a weak relationship with NO2(-) concentration (r(s) = 0.6, p = 0.1). BALF NO2(-) was strongly related to BALF % neutrophils in BOS subjects (r(s) = 0.9, p < 0.01), but there was no such relationship in STs. CONCLUSIONS: Our findings suggest that the allograft could be subject to significant iron-generated oxidative stress, which may be exacerbated by NO and neutrophil-derived ROS, particularly in BOS. Microvascular leakage may be a feature of established chronic rejection, which potentiates the iron overload and contributes to further airway damage and remodeling.

Adult↗

PubMed2001

Bronchoalveolar lavage macrophage and lymphocyte phenotypes in lung transplant recipients.

Recent publications have demonstrated potentially pathologic changes in bronchoalveolar lavage (BAL) from clinically stable lung transplant recipients (SLTRs), but there are few available data on alveolar macrophages (AMs). We formulated the hypothesis that changes in BAL AM and lymphocyte phenotypes would be apparent even in SLTRs.A cross-sectional study using a standardized 3 x 60 ml BAL, investigating lymphocyte and AM phenotypes in 19 SLTRs, 5 subjects with bronchiolitis obliterans syndrome (BOS) and 18 normal control volunteers. BAL lymphocyte and AM markers were assessed using flow cytometry. We confirmed a significant elevation of neutrophils in all lung transplant recipients with a more marked elevation in the BOS subjects. Flow-cytometric analysis showed increased numbers of natural killer (NK; CD56/CD16-positive) cells, increased CD11b- and CD11c-positive CD3 lymphocytes, increased CD8-positive lymphocytes and increased HLA-DR expression in CD8 cells from the lung transplant recipients, when compared with normals (p <.005). In contrast, the expression of a number of AM surface markers, associated with a range of host defense functions against bacteria, fungi and viruses (CD11a, CD11b, CD11c, HLA-DR, CD14), was lower in both SLTRs and those with BOS (p <.05). These novel findings are consistent with complex lymphocyte and macrophage changes that may result from clinically silent infection, partially suppressed rejection, or both.

Adult↗

PubMed2001

Studies of binaural detection in the rabbit (Oryctolagus cuniculus) with Pavlovian conditioning.

A Pavlovian conditioned eyeblink response in rabbits (Oryctolagus cuniculus) was used to study psychoacoustical phenomena previously demonstrated in human listeners and other animals. This article contains the results of a tone-in-noise detection study to examine 2 psychoacoustical phenomena in rabbit and in human listeners: (a) the binaural masking level difference (BMLD) and (b) differential performance across reproducible noise masker waveforms. The rabbits demonstrated a BMLD comparable in size to other species. Significant differences in performance across reproducible noise masker waveforms were seen in the rabbits. This performance was compared with the performance of human listeners using the same set of waveforms.

Adult↗

PubMed2001

Respiratory ciliary function in bone marrow recipients.

Bone marrow transplantation (BMT) recipients, particularly those with chronic graft-versus-host disease (GVHD), suffer from respiratory tract problems, including bronchiolitis obliterans (BO) and recurrent lower respiratory tract infections. Minute cilia beat continuously on the surface of respiratory mucosa, and this beating maintains the sterility of the lower respiratory tract. Dysfunction of respiratory cilia could lead to development of recurrent respiratory tract infections, which are also features of BMT recipients, although ciliary function has not been systematically studied among these subjects. We have, therefore, investigated the ciliary beat frequency (CBF) of 36 Chinese patients who had undergone allogeneic BMT. The CBF was significantly lower in the BMT group compared to controls (P < 0.001). The reduction in CBF was more severe in patients with cGVHD and BO compared with their counterparts (P = 0.048 and P = 0.077, respectively). There was a correlation between CBF with forced expiratory flow rate FEF (P = 0.024) and forced expiratory volume FEV (P = 0.044). We conclude that abnormal ciliary clearance is a common feature after allogeneic BMT, particularly among patients with BO and cGVHD. Further studies are indicated to evaluate this important phenomenon, which could be an important cause of the susceptibility for BMT recipients to respiratory infections.

Adolescent↗

PubMed2001

Selective determination of dopamine in the presence of ascorbic acid at an over-oxidized poly(N-acetylaniline) electrode.

An over-oxidized poly(N-acetylaniline) (PNAANI)/GCE was used to determine dopamine (DA) in a large excess of ascorbic acid (AA) by differential pulse voltammetry. A linear relation between Ip and DA concentration was found over the range 5.0 x 10(-7) to 2.0 x 10(-5) M. The detection limit was 1.68 x 10(-8) M for S/N = 3 and 400 microM AA did not interfere with the DA determination. The high sensitivity was due to accumulation and selectivity was due to charge discrimination. The mechanism of selective determination of DA at over-oxidized PNAANI/GCE was also proposed.

Ascorbic Acid↗

PubMed2001

Tehao functions in the Toll pathway in Drosophila melanogaster: possible roles in development and innate immunity.

Toll and related proteins play important roles in innate immunity in both invertebrates and vertebrates. In Drosophila melanogaster, Tehao shares a striking similarity in its intracellular domain with Toll. In this paper, we show that Tehao is expressed throughout development and appears to be glycosylated. In transiently transfected cells, Tehao activated both Dorsal and the transcription of endogenous drosomycin and metchnikowin genes. Purified recombinant Tehao interacted specifically in vitro not only with the Pelle protein kinase, but also with the Toll intracytoplasmic domain. Remarkably, Tehao was found to activate Dorsal-dependent transcription in a synergistic manner with Toll, as well as Pelle in co-transfected cells. Thus, Tehao, alone or with Toll as a multimeric complex, has the potential to participate in both the development and innate immune responses of Drosophila.

Amino Acid Sequence↗