Real-time spectroscopic techniques for probing conformational dynamics of heme proteins.
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Animals↗
Biomedical subjects
X Xie
Publications and source records attributed to X Xie.
Properties of GABA-mediated synaptic potentials induced by zinc in adult rat hippocampal pyramidal neurones.
1. Intracellular recording techniques were used to study the actions of the transition ion, zinc, on CA1 and CA3 pyramidal neurones in adult rat hippocampal slices. 2. Zinc (300 microM) hyperpolarized pyramidal neurones, increased the membrane excitability and also induced periodic, spontaneous giant depolarizing potentials associated with a conductance increase mechanism. 3. The occurrence of spontaneous giant depolarizations was dependent on the zinc concentration (10 microM-1 mM) with an apparent dissociation constant of 98 microM. The frequency of zinc-induced depolarizations was unaffected by the membrane potential from -50 to -100 mV. 4. Stimulation of the Schaffer collaterals or mossy fibre pathways evoked an excitatory and inhibitory synaptic potential complex. In the presence of zinc, nerve fibre stimulation evoked, in an all-or-none fashion, a giant depolarizing potential with an increased membrane conductance. Both spontaneous and evoked depolarizations were inhibited by 1 microM tetrodotoxin. 5. Evoked giant depolarizations were labile with too frequent stimulation resulting in a failure of generation. A minimum time of 140 s was required between stimuli to ensure successive giant depolarizations. 6. Spontaneous and evoked zinc-induced depolarizing potentials were inhibited by bicuculline (10 microM) or picrotoxin (40 microM) and enhanced by pentobarbitone (100 microM) or flurazepam (10 microM), suggesting that these potentials are mediated by activation of gamma-aminobutyric acidA (GABAA) receptors. 7. Ionophoretic application of GABA produced biphasic responses at -60 mV membrane potential. The reversal potentials for the depolarizing and hyperpolarizing GABA responses were -56 +/- 5 and -66 +/- 8 mV respectively. The giant depolarizations induced by zinc reversed at -57 +/- 4 mV. This suggests a dendritic location for the generation of these potentials. 8. Excitatory amino acid antagonists, 2-amino-5-phosphonovalerate (APV, 40 microM) or 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM) did not affect the amplitude but slightly reduced the frequency of the giant depolarizations. 9. It is concluded that zinc induces a synchronized release of GABA, quite independent of intact excitatory synaptic transmission, which acts on GABAA receptors producing large depolarizing synaptic potentials. This increased level of GABA release may be of physiological and pathological importance since zinc is a naturally occurring metal ion endogenous to the central nervous system.
Action Potentials↗
Molecular and statistical approaches to the detection and correction of errors in genotype databases.
Errors in genotyping data have been shown to have a significant effect on the estimation of recombination fractions in high-resolution genetic maps. Previous estimates of errors in existing databases have been limited to the analysis of relatively few markers and have suggested rates in the range 0.5%-1.5%. The present study capitalizes on the fact that within the Centre d'Etude du Polymorphisme Humain (CEPH) collection of reference families, 21 individuals are members of more than one family, with separate DNA samples provided by CEPH for each appearance of these individuals. By comparing the genotypes of these individuals in each of the families in which they occur, an estimated error rate of 1.4% was calculated for all loci in the version 4.0 CEPH database. Removing those individuals who were clearly identified by CEPH as appearing in more than one family resulted in a 3.0% error rate for the remaining samples, suggesting that some error checking of the identified repeated individuals may occur prior to data submission. An error rate of 3.0% for version 4.0 data was also obtained for four chromosome 5 markers that were retyped through the entire CEPH collection. The effects of these errors on a multipoint map were significant, with a total sex-averaged length of 36.09 cM with the errors, and 19.47 cM with the errors corrected. Several statistical approaches to detect and allow for errors during linkage analysis are presented. One method, which identified families containing possible errors on the basis of the impact on the maximum lod score, showed particular promise, especially when combined with the limited retyping of the identified families. The impact of the demonstrated error rate in an established genotype database on high-resolution mapping is significant, raising the question of the overall value of incorporating such existing data into new genetic maps.
Bias↗
gamma-Hydroxybutyrate depresses monosynaptic excitatory and inhibitory postsynaptic potentials in rat hippocampal slices.
The action of gamma-hydroxybutyrate was studied on pre- and postsynaptic GABA(B) receptors in rat hippocampal neurones in vitro using intracellular recording. gamma-Hydroxybutyrate (1-10 mM) caused a 4-8 mV hyperpolarization of CA1 cells and a 20-80% decrease in monosynaptic excitatory and inhibitory postsynaptic potentials in a concentration-dependent manner. These actions were reversibly inhibited by a novel and selective GABA(B) antagonist, CGP 36742 (20-500 microM) suggesting that gamma-hydroxybutyrate can activate presynaptic as well as postsynaptic GABA(B) receptors.
Animals↗
Structure of tropomyosin at 9 angstroms resolution.
We have used molecular replacement followed by a highly parameterized refinement to determine the structure of tropomyosin crystals to a resolution to 9 A. The shape, coiled-coil structure and interactions of the molecules in the crystals have been determined. These crystals have C2 symmetry with a = 259.7 A, b = 55.3 A, c = 135.6 A and beta = 97.2 degrees. Because of the unusual distribution of intensity in X-ray diffraction patterns from these crystals, it was possible to solve the rotation problem by inspection of qualitative aspects of the diffraction data and to define unequivocally the general alignment of the molecules along the (332) and (3-32) directions of the unit cell. The translation function was then solved by a direct search procedure, while electron microscopy of a related crystal form indicated the probable location of molecular ends in the asymmetric unit, as well as the anti-parallel arrangement. The structural model we have obtained is much clearer than that obtained previously with crystals of extraordinarily high solvent content and shows the two alpha-helices of the coiled coil over most of the length of the molecules and establishes the coiled-coil pitch at 140(+/- 10) A. Moreover, the precise value of the coiled-coil pitch varies along the molecule, probably in response to local variations in the amino acid sequence, which we have determined by sequencing the appropriate cDNA. The crystals are constructed from layers of tropomyosin filaments. There are two molecules in the crystallographic asymmetric unit and the molecules within a layer are bent into an approximately sinusoidal profile. Molecules in consecutive layers in the crystal lie at an angle relative to one another as found in crystalline arrays of actin and myosin rod. There are three classes of interactions between tropomyosin molecules in the spermine-induced crystals and these give some insights into the molecular interactions between coiled-coil molecules that may have implications for assemblies such as muscle thick filaments and intermediate filaments. In interactions within a layer, the geometry of coiled-coil contacts is retained, whereas in contacts between molecules in adjacent layers the coiled-coil geometry varies and these interactions instead appear to be dominated by the repeating pattern of charged zones along the molecule.
Amino Acid Sequence↗
Femtosecond spontaneous-emission studies of reaction centers from photosynthetic bacteria.
Spontaneous emission from reaction centers of photosynthetic bacteria has been recorded with a time resolution of 50 fs. Excitation was made directly into both the special-pair band (850 nm) and the Qx band of bacteriochlorophylls (608 nm). Rhodobacter sphaeroides R26, Rhodobacter capsulatus wild type, and four mutants of Rb. capsulatus were studied. In all cases the fluorescence decay was not single exponential and was well fit as a sum of two exponential decay components. The short components are in excellent agreement with the single component detected by measurements of stimulated emission. The origin of the nonexponential decay is discussed in terms of heterogeneity, the kinetic scheme, and the possibility of slow vibrational relaxation.
Energy Transfer↗
Strained alpha -Sn/Ge superlattices: Geometrical structures and phonon spectra.
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Journal Article↗
Phonon spectra of strained Si and Ge.
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Journal Article↗
Gamma-hydroxybutyrate hyperpolarizes hippocampal neurones by activating GABAB receptors.
gamma-Hydroxybutyrate, a naturally occurring substance present in the mammalian central nervous system caused a dose-dependent (0.25-10 mM) hyperpolarization and small membrane conductance increase when applied to hippocampal CA1 pyramidal neurones in vitro. This action was reversibly inhibited by the GABAB antagonist, CGP 35348 (20-100 microM) and divalent cations, Zn2+ and Ba2+, but not by the GABAA antagonist bicuculline (50 microM). These results suggest that GABAB receptors may mediate the actions of gamma-hydroxybutyrate.
Animals↗
Comparative studies between nude and scid mice on the growth and metastatic behavior of xenografted human tumors.
The growth and metastatic behavior of three human tumor cell lines and a human colon carcinoma previously passaged in vivo were compared between nude mice and scid mice after xenotransplantation. The three human tumor lines included a bladder carcinoma (T24B), a melanoma (RPMI 7931) and a lacZ gene-transduced breast cancer (MDA-MB-435 BAG). The lacZ gene codes for beta-galactosidase, which can be stained blue with chromogenic substrate X-gal, thus allowing the highly sensitive detection and quantitative examination of human cancer metastasis in host mice. Adult (7-14 weeks) NMRI nude and C.B-17 SCID mice were inoculated with 0.5-5 x 10(6) tumor cells s.c. Comparable take rate, latent period and growth rate of implanted tumors were observed in nude and scid mice for each of the cell lines tested. At the time of autopsy, which varied from 6 to 11 weeks after inoculation, a significantly higher incidence of spontaneous lung metastasis was discovered in scid mice (96%) than in age-matched nude mice (27%, total P less than 0.001). In vitro assays for NK cell-mediated cytotoxicity revealed no significant differences between the two strains of mice. Our results suggest that nude and scid mice are equally suitable for propagating human tumors. However, the metastatic capacity of human tumor cells appears to be better expressed in scid mice. Scid mice may therefore provide an advantageous model for the study of human tumor metastasis.
Animals↗
Chemoattractant-induced firm adhesion of leukocytes to vascular endothelium in vivo is critically dependent on initial leukocyte rolling.
Leukocyte rolling and firm adhesion at the venular endothelium are two discrete events in the cellular inflammatory response mediated via selectin and integrin adhesion molecules, respectively. The dependency of chemoattractant-induced firm leukocyte adhesion on the preceding rolling interaction was investigated in rat mesenteric microvessels through use of intravital microscopy. Leukocyte rolling was dose-dependently inhibited by systemic treatment with the sulphated polysaccharide fucoidin. The firm leukocyte adhesion following stimulation with the chemotactic peptide fMLP was similarly inhibited when fMLP challenge was performed subsequent to inhibition of leukocyte rolling by fucoidin. Thus, based on paired observations in single venules before and after fucoidin treatment, reduced rolling leukocyte flux prior to fMLP challenge was paralleled over a wide range by a proportional decrease in fMLP-induced leukocyte adhesion. The results demonstrate quantitatively a close relationship between the extent of leukocyte rolling and the magnitude of the subsequent firm adhesion response, and, that an initial rolling interaction is a precondition for firm adhesion to occur at physiological blood flow rates in vivo.
Animals↗
Isolated NMDA receptor-mediated synaptic responses express both LTP and LTD.
1. The possibility of use-dependent, long-lasting modifications of pharmacologically isolated N-methyl-D-aspartate (NMDA) receptor-mediated synaptic transmission was examined by intracellular recordings from granule cells of the hippocampal dentate gyrus in vitro. In the presence of the non-NMDA receptor antagonist 6-cyano-7-nitroquinaxaline-2,3-dione (CNQX, 10 microM) robust, long-term potentiation (LTP) of NMDA receptor-mediated synaptic potentials was induced by brief, high (50 Hz) and lower (10 Hz) frequency tetanic stimuli of glutamatergic afferents (60 +/- 6%, n = 8, P less than 0.001 and 43 +/- 12%, n = 3, P less than 0.05, respectively). 2. Hyperpolarization of granule cell membrane potential to -100 mV during 50-Hz tetanic stimuli reversibly blocked the induction of LTP (-6 +/- 2%, n = 6, P greater than 0.05) indicating that simultaneous activation of pre- and postsynaptic elements is a prerequisite for potentiation of NMDA receptor-mediated synaptic transmission. In contrast, hyperpolarization of the granule cell membrane potential to -100 mV during 10-Hz tetanic stimuli resulted in long-term depression (LTD) of NMDA receptor-mediated synaptic potentials (-34 +/- 8%, n = 8, P less than 0.01). 3. We also studied the role of [Ca2+]i in the induction of LTP and LTD of NMDA receptor-mediated synaptic responses. Before tetanization, [Ca2+]i was buffered by iontophoretic injections of bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA). BAPTA completely blocked the induction of LTP (3 +/- 5%, n = 13) and partially blocked LTD (-14.8 +/- 6%, n = 10).(ABSTRACT TRUNCATED AT 250 WORDS)
2-Amino-5-phosphonovalerate↗
[Hepatic carcinoma treated by hepatic arterial embolization using 131I and chemotherapeutic agent gelatin microspheres: report of 9 cases].
Nine patients with inoperable hepatoma were treated by using hepatic arterial embolization 131I and chemotherapeutic agent gelatin microsphere (131I-CA-GM). The emission CT after operation detected that the microspheres were concentrated on tumor area. The ratio between the radioactivity in tumor and that in liver was 4.1:1. A case died of ictopic embolization; the others survived 3, 4, 5, 19, 24, 7, 8, and 12 months respectively. Three of them were still alive. 131I-CA-GM has triple anticarcinogenic actions, including the arterial occlusion, targeting chemotherapy and internal radiation. The microspheres can selectively accumulate in the tumor artery and can be easily traced by gamma-camera or emission CT. 131I-CA-GM is a hopeful embolic agent for the treatment of liver cancer, but some problems about ectopic arterial embolization should be further studied.
Carcinoma, Hepatocellular↗
Molecular characterization and structural organization of D-elg, an ets proto-oncogene-related gene of Drosophila.
We have continued the molecular analysis of D-elg, a member of the Drosophila ets gene family. Based on the characterization of cDNA and genomic sequences, the D-elg gene contains five exons and four introns and produces a mRNA with an open reading frame of 464 amino acids. Consistent with this analysis, in vitro translation of a near full-length D-elg cRNA yields a protein with a molecular weight of approximately 56 kDa. D-elg shows significant homology to other ets proteins in the amino-terminal A domain and strong homology in the carboxy-terminal ETS domain. The D-elg protein is most similar to the alpha-subunit of the mouse GA-binding protein.
Amino Acid Sequence↗
Endoderm-specific expression of the Drosophila mex1 gene.
The Drosophila mex1 gene is one of several genes clustered within a 10-kb interval of polytene region 71CD that includes the ecdysone-regulated Eip28/29 gene. mex1 is expressed in several developmental stages, with gene transcripts accumulating initially in 9- to 12-hr embryos. During embryogenesis, mex1 exhibits an endoderm-specific pattern of expression. mex1 transcripts are first detected in the anterior and posterior midgut primordia of stage 12 embryos; subsequently, mex1 mRNA accumulates solely in the differentiating embryonic midgut. DNA sequence analysis reveals that the mex1 gene encodes an unusual cysteine-rich polypeptide.
Amino Acid Sequence↗
GABAA receptors are differentially sensitive to zinc: dependence on subunit composition.
GABAA receptors with different subunit composition, were expressed in kidney cells and studied by whole cell recording. Expressed GABAA receptors were differentially sensitive to inhibition by zinc; receptors which lacked the gamma subunit were inhibited by zinc. Embryonic neurons also exhibited zinc-sensitive GABA responses, in contrast to adult neurones. This developmentally-sensitive aspect of GABAA receptor pharmacology may be partly dependent on expression of the gamma subunit.
Animals↗
[Opioid peptides in cerebrospinal fluids of epileptic patients].
By means of RIA, the contents of Leu-enkephalin, Met-enkephalin, and Beta-endorphin in CSF of 32 epileptic patients and 24 controls were determined. It was found that the mean Leu-enkephalin content in CSF of the epileptic patient group was significantly higher than that of the control group (P less than 0.01), whereas the mean contents of Met-enkephalin and Beta-endorphin in CSF showed no significant change as compared with those of the control group. The increase of Leu-enkephalin was not related to such factors as type of seizure, age of onset, length of time after the last seizure, taking of antiepileptic drugs, and abnormality in cranial CT manifestation. This suggested that endogenous opioid peptides might take part in the neurochemical mechanism of human epilepsy, and leu-enkephalin could play an important role in the development of epileptic episodes.
Adolescent↗
[Assay of S-100+ Langerhans cells in cervical carcinoma].
Langerhans cells (LC) in the normal cervix (control, 19 cases) and cervical carcinoma in situ (CIS, 19 cases), interdigitating cells (IDC) and follicular dendritic cells (FDC) in obturator lymph node draining infiltrating cervical carcinoma (30 cases) were quantitatively assayed by ABC immunohistochemical method using anti-S-100 protein antibody. The results indicated that S-100+ LC in situ, S-100+ IDC and S-100+ FDC in obturator lymph node showed dendritic features with a specific distribution. The number of LC in situ in infiltrating carcinoma increased significantly as compared with CIS and control groups. There was no change in LC number between Grades II and III of squamous cell carcinoma. The number of IDC was significantly less in Stage II than in Stage I lesion. FDC number in different depths of invasion and clinical stages showed no obvious change. The results suggest that progression of cervical carcinoma is closely related to decrease in LC in situ and IDC in regional lymph node. Predominant immune response of the host to cervical carcinoma should be cellular immune.
Antibodies↗