Creutzfeldt-Jakob disease in a Chinese patient with a novel seven extra-repeat insertion in PRNP.
Explore the source record for details and available documents.
Adult↗
Biomedical subjects
X F Wang
Publications and source records attributed to X F Wang.
Charge transfer via a two-strand superexchange bridge in DNA.
Charge transfer in a DNA duplex chain is studied by constructing a system with virtual electrodes connected at the ends of each DNA strand. The system is described by the tight-binding model and its transport is analyzed by the transfer matrix method. The very weak distance dependence in a long (G:C)(T:A)M(G:C)3 DNA chain observed in experiment [B. Giese, Nature (London) 412, 318 (2001)] is explained by a unistep two-strand superexchange bridge without the need for the multistep thermally induced hopping mechanism or the dephasing effect. The crossover number Mc of the (T:A) base pairs, where crossover between the strong and weak distance dependence occurs, reflects the ratio of intra- and interstrand neighboring base-base couplings.
Journal Article↗
[Significance of hepatitis B virus PreS1-Ag, PreS2-Ag, large protein, PreS2-Ab detection and the prediction of HBV DNA replication.].
BACKGROUND: To explore the significance of hepatitis B virus PreS1-Ag, PreS2-Ag, large protein (LP) detection and the prediction of viral replication. METHODS: PreS1-Ag, PreS2-Ag, LP and HBV markers were measured by enzyme linked immunosorbent assay (ELISA) in 201 cases of infected serum. Serum HBV DNA level was quantitatively detected by real-time polymerase chain reaction (PCR). RESULTS: There were significant differences in positive rate between the PreS1-Ag, PreS2-Ag, LP, and HBsAg; the positive rate of PreS2-Ag and LP were higher than that of the HBeAg. No significant differences were found in the positive rates between LP and the levels of HBV DNA and there was a positive correlation between quantitations of HBV DNA and HBV-LP. CONCLUSION: Serum PreS1-Ag, PreS2-Ag and LP were laboratory markers that can accurately reflect HBV DNA reproduction, and were helpful complementarity to traditional HBV M. There is a close correlation between the number of copies of HBV DNA and the levels of HBV-LP.
DNA Replication↗
Normal CFTR Activity and Reversed Skin Potentials in Pseudohypoaldosteronism.
Cystic fibrosis (CF) transmembrane conductance regulator (CFTR) Cl(-) channel function is required for activating amiloride-sensitive epithelial Na(+) channels (ENaC) in salt-absorbing human sweat duct. It is unclear whether ENaC channel function is also required for CFTR activation. The dysfunctional ENaC mutations in type-1 pseudohypoaldosteronism (PHA-1) provided a good opportunity to study this phenomenon of ion channel interaction between CFTR and ENaC. The PHA-1 ducts completely lacked spontaneous ENaC conductance (gENaC). In contrast, the normal ducts showed large spontaneous gENaC (46 +/- 10 ms, mean +/- SE: ). After permeabilization of the basolateral membrane with alpha-toxin, cAMP + ATP activation of CFTR Cl(-) conductance (gCFTR) or alkalinization of cytosolic pH (6.8 to 8.5) stimulated gENaC of normal but not PHA-1 ducts. In contrast, both spontaneous gCFTR in intact ducts and (cAMP + ATP)-activated gCFTR of permeabilized ducts appeared to be similar in normal and PHA-1 subjects. Lack of gENaC completely blocked salt absorption and caused dramatic reversal of skin potentials associated with pilocarpine-induced sweat secretion from significantly negative in normal subjects (-13 +/- 7.0 mV) to significantly positive (+22 +/- 11.0 mV) in PHA-1 patients. We conclude that virtual lack of ENaC in PHA-1 ducts had little effect on CFTR activity and that the positive skin potentials could potentially serve as a diagnostic tool to identify type-1 pseudohypoaldosteronism.
Cystic Fibrosis Transmembrane Conductance Regulato↗
Effects of a new cystic fibrosis transmembrane conductance regulator inhibitor on Cl- conductance in human sweat ducts.
Effective and specific inhibition of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel in epithelia has long been needed to better understand the role of anion movements in fluid and electrolyte transport. Until now, available inhibitors have required high concentrations, usually in the millimolar or high micromolar range, to effect even an incomplete block of channel conductance. These inhibitors, including 5-nitro-2(3-phenylpropyl-amino)benzoate (NPPB), bumetamide, glibenclamide and DIDS, are also relatively non-specific. Recently a new anion channel inhibitor, a thiazolidinone derivative, termed CFTRInh-172 has been synthesized and introduced with apparently improved inhibitory properties as shown by effects on anion conductance expressed in cell lines and on secretion in vivo. Here, we assay the effect of this inhibitor on a purely salt absorbing native epithelial tissue, the freshly isolated microperfused human sweat duct, known for its inherently high expression of CFTR. We found that the inhibitor at a maximum dose limited by its aqueous solubility of 5 microm partially blocked CFTR when applied to either surface of the membrane; however, it may be somewhat more effective from the cytosolic side (approximately 70% inhibition). It may also partially inhibit Na+ conductance. The inhibition was relatively slow, with a half time for maximum effect of about 3 min, and showed very slow reversibility. Results also suggest that CFTR Cl- conductance (GCl) was blocked in both apical and basal membranes. The inhibitor appears to exert some effect on Na+ transport as well.
Benzoates↗
Long term potentiation varies with layer in rat visual cortex.
Long term potentiation (LTP) in various layers of rat visual cortex was studied in 90 cells with visually identified, whole-cell recordings. LTP was induced in layer II/III, layer V or layer VI with theta burst stimulation (TBS), but was not observed in layer IV. In the presence of a NMDA antagonist, D-AP5, in the bath solution, potentiation was blocked in layer II/III, some depression was seen in layer V, and potentiation still remained in layer VI. After addition of a specific mGluR1 antagonist, LY367385, to the bath solution, LTP was reduced in layer II/III and layer V, and was blocked in layer VI. After a specific mGluR5 antagonist, MPEP was applied in the bath solution, LTP was enhanced in layer VI, and blocked in layer V. We conclude that: (1) LTP in layer VI is different from other layers, depending on mGluR1, but not NMDA receptors. (2) In layer II/III, LTP is NMDA-dependent and is not blocked by group I mGluR antagonists. (3) LTP in layer V is both NMDA receptor and mGluR5 receptor-dependent. (4) LTP was not induced in layer IV with TBS.
Animals↗
Electrical properties affecting discharge of units of the mid and posterolateral thalamus of conscious cats.
Discharge properties in response to intracellularly applied, rectangular currents were measured in units of the mid (lateralis dorsalis and centrolateral nuclei) and posterolateral (lateralis posterior and pulvinar nuclei) thalamus of conscious cats. A separate aim was to determine if neuronal excitability changed in association with changes in stimulus-evoked activity after the animals were trained to discriminate between two acoustic stimuli when performing a conditioned motor response. Low threshold spike (l.t.s.) discharges were observed in three of 272 cells given 1 nA intracellular, hyperpolarizing current pulses of 40 ms duration. This finding supports the view that thalamic neurons of conscious animals operate mainly in the relay as opposed to the oscillatory mode. Application of larger and longer hyperpolarizing currents in the cells produced rebound l.t.s. discharges, supporting the expectation that most thalamic neurons are capable of producing this type of discharge. Decrements of spike afterhyperpolarizations (AHP) and broadening of spike bases upon repeated discharge also were observed in each area of the thalamus studied. After conditioning, changes were found in the posterolateral thalamus (but not in the mid-thalamus) in the proportions of cells with spontaneous, rapid (>/=50 Hz), repetitive, discharges (RRD) and rapid, sustained discharges at rates >/=100 Hz during application of depolarizing current (RSD). In the posterolateral thalamus the percentage of units responding to 1 nA depolarization with RSD fell from 71% before conditioning to 45% after conditioning. The percentage of cells with RRD decreased from 69% to 46%. The changes were accompanied by a 3 mV hyperpolarization of the membrane potentials of the cells and a decrease in baseline activity. After conditioning, increases in excitability were found in cells of the mid thalamus that responded selectively to the click conditioned stimulus (CS) that elicited the conditioned response, and decreases in excitability were found in cells of the posterolateral thalamus that responded to the discriminative acoustic stimulus (DS) to which the animals were trained not to respond. An earlier study showed a potentiation of discharge in response to the CS in units of the midthalamus after similar conditioning and a reduction of the proportion of DS responsive units and peak discharge to the DS in units of the posterolateral thalamus. We conclude that the discharge properties of units of the mid and posterolateral thalamus can change to support discrimination between acoustic stimuli of different functional significance after conditioning.
Acoustic Stimulation↗
Expression, immunolocalization, and functional activity of Na+/H+ exchanger isoforms in mouse endometrial epithelium.
The luminal fluid microenvironment of the uterus is important for sperm capacitation and embryo development. In an attempt to understand the possible role of Na(+)/H(+) exchangers (NHEs) in uterine function, the mRNAs of different NHE isoforms as well as their subcellular localization (apical versus basolateral) and functional activity were investigated in mouse endometrial epithelial cells using reverse transcriptase-polymerase chain reaction (RT-PCR), immunohistochemistry, and intracellular pH (pH(i)) measurement techniques. The presence of NHE1, NHE2, and NHE4, but not NHE3 mRNAs were revealed by RT-PCR. Immunostaining showed that NHE1, NHE2, and NHE4 were present in both apical and basolateral membranes. The pH(i) recovery from intracellular acidification was Na(+)-dependent; however, the rate of pH(i) recovery depending on basolateral Na(+) was 12.4 times faster than that depending on apical Na(+). The Na(+)-dependent rate of pH(i) recovery was also inhibited by amiloride, indicating H(+) extrusion through NHEs; however, the amiloride sensitivity of the apical membrane was less than that of the basolateral membrane, suggesting the involvement of different types of NHEs in the two membranes. The results indicate that the basolaterally located NHE1, NHE2, and NHE4, in addition to participating in the homeostatic control of intracellular pH, may play a role in H(+) extrusion in order to achieve transepithelial HCO(3)(-) secretion. The apically located NHEs may be involved in mediating Na(+) absorption as alternatives of or complementary to epithelial Na(+) channels.
Animals↗
Clinical and immunohistochemical evidence for an X linked retinitis pigmentosa syndrome with recurrent infections and hearing loss in association with an RPGR mutation.
Explore the source record for details and available documents.
Adolescent↗
Transforming growth factor beta 1 dysregulation in a human oral carcinoma tumour progression model.
A human oral tumour progression model was established that consists of normal epithelial cells and three cell lines representing stages from dysplastic to metastatic cells. To investigate the impact of exogenous transforming growth factor-beta 1 on this model system, we analysed the responsiveness of those cells to transforming growth factor-beta 1 and explored the potential mechanism underlying the transforming growth factor-beta 1 activity. We found that the growth of all cell types, regardless of their stage of tumour progression, is inhibited by transforming growth factor-beta 1, although to different degrees. Transforming growth factor-beta 1 induced the expression of cyclin-dependent kinase inhibitors p15(INK4B), p21WAF1/(CIP1) and p27(KIP1). In contrast, transforming growth factor-beta 1 was found to stimulate the invasive potential of one cell type that represents the most advanced stage of tumour phenotype, suggesting that the impact of transforming growth factor-beta 1 on functional features of tumour cells other than cellular proliferation may play a significant role in the process of oral tumour progression.
Active Transport, Cell Nucleus↗
Involvement of Na+-HCO3- cotransporter in mediating cyclic adenosine 3',5'-monophosphate-dependent HCO3- secretion by mouse endometrial epithelium.
The present study investigated the involvement of Na+-HCO3- cotransporter in mediating cAMP-stimulated HCO3- secretion across the cultured mouse endometrial epithelium using the short-circuit current (I(SC)) technique and intracellular pH measurement. Forskolin stimulated a rise in the I(SC), 55.6% and 52.1% of which could be reduced by the removal of extracellular Cl- or by eliminating the contribution of Cl- secretion by bumetanide, an inhibitor of Na+-K+-2Cl- cotransporter, respectively. More than 80% reduction in the forskolin-induced I(SC) was obtained when both Cl- and HCO3- in the bath were removed or in HCO3--free solution with bumetanide, indicating that the I(SC) depended on both Cl- and HCO3-. The presence of the Na+ channel-blocker amiloride in the apical solution did not reduce the forskolin-induced I(SC); however, the I(SC) could be abolished by removing Na+ from the bathing solution, suggesting that the Cl-- and HCO3--dependent I(SC) was also dependent on basolateral Na+. The forskolin-stimulated I(SC) could be reduced 43.6% by removal of HCO3- and 47.9% by a Na+-HCO3--cotransporter inhibitor, dihydrogen-4,4'-didsothiocyanostilbene-2,2'-disulfonic acid (H2DIDS). The inhibitory effect of H2DIDS was observed in Cl--free solution, but not when HCO3- was removed, thus confirming its effect on HCO3--dependent transport. Intracellular pH measurements demonstrated that the recovery from cellular acidification depended on the presence of both basolateral Na+ and HCO3-, further indicating the involvement of Na+-HCO3- cotransporter. Reverse transcription-polymerase chain reaction experiments confirmed the expression of Na+-HCO3- cotransporter in the mouse endometrium. The results suggest that basolaterally located Na+-HCO3- cotransporter is involved in mediating cAMP-stimulated HCO3- secretion across the mouse endometrial epithelium.
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
Synergistic outside-in regulation of platelet activation by GPIIb/IIIa ligand-induced conformation and oligomerization.
Full platelet activation with serotonin secretion and thromboxane A(2) (TxA(2)) formation induced by a low dose of thrombin receptor agonist peptide (TRAP) or high dose ADP requires platelet aggregation. This requirement can be replaced by pretreatment of platelets with a combination of reagents including: GPIIb/IIIa inhibitors yielding ligand-induced binding sites (LIBS), either arginine-glycine-aspartate-serine (RGDS) peptide or Ro 43-5054, cytochalasin to disrupt actin filaments and crosslinking by a GPIIb/IIIa mAb (pl-62). Crosslinking is required since Fab fragments of pl-62 do not support activation. Engagement of the Fc receptor by the mAb Fc domain is not required for pl-62 augmentation, since it is not blocked by the anti-Fc receptor mAb, IV-3. Another GPIIb/IIIa inhibitor, Ro 44-9883, not yielding LIBS epitopes, serves as a negative control and shows a requirement for LIBS in addition to crosslinking. Focal adhesion kinase tyrosine phosphorylation induced by TRAP is blocked by these GPIIb/IIIa antagonists, but restored by pl-62 crosslinking independent of LIBS induction. Tyrosine phosphorylation of a peptide comigrating with p38 MAP kinase is also inhibited by these antagonists and restored by pl-62 crosslinking. However, p38 MAP kinase activation by low dose TRAP is not affected by these aggregation inhibitors. Tyrosine phosphorylation of a 34-kDa phosphoprotein in the absence of aggregation or TxA(2) formation was uniquely augmented by Ro 43-5054 but not Ro 44-9883 under the above activation conditions.
Acetates↗
Inhibition of amiloride-sensitive Na(+) absorption by activation of CFTR in mouse endometrial epithelium.
Previous studies have demonstrated amiloride-sensitive Na(+) absorption under basal conditions and cystic fibrosis transmembrane conductance regulator (CFTR)-mediated Cl(-) secretion following neurohormonal stimulation in the mouse endometrial epithelium. The present study investigated the inhibition of amiloride-sensitive Na(+) absorption accompanying activation of CFTR in the mouse endometrium using the short-circuit current ( I(sc)) technique. RT-PCR demonstrated the co-expression of CFTR and epithelial Na(+) channels (ENaC) in primary cultured mouse endometrial epithelia and cultured endometrial monolayers exhibited a basal amiloride-sensitive I(sc) of 5.4 +/- 0.6 microA/cm(2). The amiloride-sensitive current fell to 3.1 +/- 0.5 microA/cm(2) after stimulation with forskolin. When the possible contribution of Na(+) absorption to the I(sc) was eliminated by amiloride (1 microM) or Na(+) replacement, the forskolin-induced I(sc) was not reduced, but rather increased significantly compared with that in the absence of amiloride or in Na(+)-containing solutions ( P < 0.02), indicating that the forskolin-induced I(sc) was mediated by Cl(-) secretion, portion of which may be masked by concurrent inhibition of basal Na(+) absorption if the contribution of Na(+) is not eliminated. When the contribution of Cl(-) to the I(sc) was eliminated by diphenylamine 2,2'-dicarboxylic acid (DPC, 2 mM) or Cl(-) replacement, forskolin now decreased, rather than increased the I(sc), demonstrating the inhibition of Na(+) absorption upon stimulation. Our data suggest an interaction between CFTR and ENaC, which may be the underlying mechanism for balancing Na(+) absorption and Cl(-) secretion across the mouse endometrial epithelium.
Amiloride↗
ATR/ATM-mediated phosphorylation of human Rad17 is required for genotoxic stress responses.
Genotoxic stress triggers the activation of checkpoints that delay cell-cycle progression to allow for DNA repair. Studies in fission yeast implicate members of the Rad family of checkpoint proteins, which includes Rad17, Rad1, Rad9 and Hus1, as key early-response elements during the activation of both the DNA damage and replication checkpoints. Here we demonstrate a direct regulatory linkage between the human Rad17 homologue (hRad17) and the checkpoint kinases, ATM and ATR. Treatment of human cells with genotoxic agents induced ATM/ATR-dependent phosphorylation of hRad17 at Ser 635 and Ser 645. Overexpression of a hRad17 mutant (hRad17AA) bearing Ala substitutions at both phosphorylation sites abrogated the DNA-damage-induced G2 checkpoint, and sensitized human fibroblasts to genotoxic stress. In contrast to wild-type hRad17, the hRad17AA mutant showed no ionizing-radiation-inducible association with hRad1, a component of the hRad1-hRad9-hHus1 checkpoint complex. These findings demonstrate that ATR/ATM-dependent phosphorylation of hRad17 is a critical early event during checkpoint signalling in DNA-damaged cells.
Animals↗
Pyruvate released by astrocytes protects neurons from copper-catalyzed cysteine neurotoxicity.
We have found previously that astrocytes can provide cysteine to neurons. However, cysteine has been reported to be neurotoxic although it plays a pivotal role in regulating intracellular levels of glutathione, the major cellular antioxidant. Here, we show that cysteine toxicity is a result of hydroxyl radicals generated during cysteine autoxidation. Transition metal ions are candidates to catalyze this process. Copper substantially accelerates the autoxidation rate of cysteine even at submicromolar levels, whereas iron and other transition metal ions, including manganese, chromium, and zinc, are less efficient. The autoxidation rate of cysteine in rat CSF is equal to that observed in the presence of approximately 0.2 microm copper. In tissue culture tests, we found that cysteine toxicity depends highly on its autoxidation rate and on the total amount of cysteine being oxidized, suggesting that the toxicity can be attributed to the free radicals produced from cysteine autoxidation, but not to cysteine itself. We have also explored the in vivo mechanisms that protect against cysteine toxicity. Catalase and pyruvate were each found to inhibit the production of hydroxyl radicals generated by cysteine autoxidation. In tissue culture, they both protected primary neurons against cysteine toxicity catalyzed by copper. This protection is attributed to their ability to react with hydrogen peroxide, preventing the formation of hydroxyl radicals. Pyruvate, but not catalase or glutathione peroxidase, was detected in astrocyte-conditioned medium and CSF. Our data therefore suggest that astrocytes can prevent cysteine toxicity by releasing pyruvate.
Animals↗
A genetically tractable model of human glioma formation.
Gliomas remain one of the deadliest forms of cancer. Improved therapeutics will require a better understanding of the molecular nature of these tumors. We, therefore, mimicked the most common genetic changes found in grade III-IV gliomas, disruption of the p53 and RB pathways and activation of telomere maintenance and independence from growth factors, through the ectopic expression of the SV40 T/t-Ag oncogene, an oncogenic form of H-ras (H-ras(V12G)), and the human telomerase catalytic subunit hTERT in normal human astrocytes. The resulting cells displayed many of the hallmarks of grade III-IV gliomas, including greatly expanded life span and growth in soft agar and, most importantly, were tumorigenic with pathology consistent with grade III-IV neuroectodermal tumors in mice. This model system will, for the first time, allow the biological significance of selected genetic alterations to be studied in human gliomas.
Animals↗
An essential role for Mad homology domain 1 in the association of Smad3 with histone deacetylase activity*.
The Smads are a family of sequence-specific DNA-binding proteins that modulate transcription in response to transforming growth factor beta (TGFbeta) by recruiting transcriptional activators like the histone acetyltransferase, p300/CBP, or repressors like the histone deacetylase, HDAC1, to TGFbeta target genes. The association of Smads and HDAC1 is mediated in part by direct binding of Smads to the HDAC1-associated proteins, TG-interacting factor, c-ski, and SnoN. Although ectopic expression of these proteins inhibits Smad-activated transcription, the contribution of histone deacetylase enzymatic activity to transcriptional repression by TGFbeta is unknown. Here, the biological requirements for the interaction between Smads and endogenous histone deacetylase activity are investigated. We identify residues in Mad homology domain 1 of Smad3 that are required for association with histone deacetylase activity. An amino acid change at one of these critical residues does not disrupt the association of Smad3 with c-ski, SnoN, and transforming growth-interacting factor but does abrogate the ability of Smad3 to repress transcription. These findings indicate that the association of Smad3 and histone deacetylase activity relies on additional protein mediators that make contact with Smad3 at its amino terminus. Moreover, these data suggest that the suppressive effect of Smad3 on transcription is dependent upon its association with histone deacetylase enzymatic activity.
Base Sequence↗
The activity of guanine exchange factor NET1 is essential for transforming growth factor-beta-mediated stress fiber formation.
To examine signaling pathways underlying transforming growth factor-beta (TGF-beta)-mediated changes in cell morphology, we used a microarray system to identify downstream target genes that may play a role in this process. Through this approach, we found that the NET1 gene was induced upon TGF-beta treatment in several cell types. NET1 is a guanine nucleotide exchange factor for RhoA whose activity has been implicated in stress fiber formation. In the Swiss 3T3 cell line, TGF-beta induces NET1 expression, and this correlated with an increase in stress fiber formation. Overexpression of the wild type NET1 gene increases stress fiber formation, and overexpression of a dominant negative NET1 mutant (L392E) prevented TGF-beta dependent increase in stress fiber formation. Furthermore, treatment of the cells with a RhoA kinase inhibitor Y-27632 blocks TGF-beta-induced stress fiber formation. By using a stable cell line expressing dominant negative Smad3, we found that the Smad signaling pathway is essential for the induction of NET1, which in turn leads to the increase of Rho activity. Taken together, those data suggest that induction of NET1 is important for the increase of Rho activity upon TGF-beta treatment, which may represent the critical trigger for a variety of downstream events in different cells. Our results support the presence of a novel signaling pathway by which TGF-beta may regulate the formation of stress fibers and reorganization of cytoskeletal structures.
3T3 Cells↗