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Biomedical subjects

H Liu

Publications and source records attributed to H Liu.

At least 127 records · Page 7Linked to original sources

Expression patterns of 5-HT7 receptor isoforms in the rat digestive tract.

In this study, we determined the expression patterns of the 5-HT7 receptor isoforms at the mRNA level in the rat jejunum and other regions of the rat digestive tract using reverse transcriptase-PCR. In control studies using the rat thalamus, we detected all three known isoforms of the rat 5-HT7 receptor, 5-HT7(a), 5-HT7(b) and 5-HT7(c). In addition, we also detected in the thalamus a new PCR product that we provisionally name 5-HT7(e). All four isoforms of the 5-HT7 receptor were present throughout the rat digestive tract. However, the spatial distribution and levels of expression of the different isoforms varied between regions of the rat digestive tract. All three known isoforms and the provisional isoform were found in the jejunum, ileum, stomach fundus, oesophagus and colon. The level of expression of the isoforms varied with all four being most consistently found in the ileum. The 5-HT7(a) receptor isoform was predominantly expressed in the rat digestive tissue and 5-HT7(b,, 5-HT7(c) and 5-HT7(e) were only sometimes expressed but always accompanying the receptor isoform 5-HT7(a).

Animals↗

DNA array studies demonstrate convergent regulation of virulence factors by Cph1, Cph2, and Efg1 in Candida albicans.

Candida albicans, normally a human commensal, can cause fatal systemic infections under certain circumstances. Its unique ability to switch from yeast to hyphal growth in response to various environmental signals is inherent to its pathogenicity. Filamentation is regulated by multiple pathways including a Cph1-mediated mitogen-activated protein kinase pathway, an Efg1-mediated cAMP/PKA pathway, and a Cph2 pathway. To gain a general picture of how these various signaling pathways regulate differential gene expression during filamentation, we have constructed a partial C. albicans DNA array of 7,000 genes and used it to study the gene expression profiles using various mutants and growth conditions. By combining this novel technology with a new liquid medium in which cph1/cph1 is defective in filamentation, previously identified differentially expressed genes (ECE1, HWP1, HYR1, RBT1, SAPs5-6, and RBT4) are found to be regulated by all three pathways. In addition, two novel genes, DDR48 and YPL184, have been found to be differentially regulated during hyphal development and by all three pathways. This suggests that distinct filamentation signaling pathways converge to regulate a common set of differentially expressed genes. As one of the mechanisms for the observed convergence, we find that the transcription of a key regulator, TEC1, is regulated by Efg1 and Cph2. Importantly, most of the genes regulated by multiple filamentation pathways encode known virulence factors. Perhaps, C. albicans utilizes converging pathways to regulate its vital virulence factors to ensure its survival and pathogenicity in various host environments.

Basic Helix-Loop-Helix Proteins↗

Molecular classification of estrogens.

Estrogens are involved in a multiplicity of programmed events in target tissues e.g.: uterus, breast, and pituitary gland, and hormone-responsive tumors occur at these target sites. We have addressed the possibility that all of the estrogens do not produce the same conformation of estrogen receptor alpha (ER). A novel assay in vitro was used to activate the transforming growth factor alpha (TGF-alpha) gene in situ in MDA-MB-231 cells stably transfected with cDNA for D351 ER or D351G ER. Three estrogen types were used: estradiol, diethylstilbestrol, and a triphenylethylene (TPE) derivative of tamoxifen without the antiestrogenic side chain. Computer molecular modeling was used to interpret data. A flat estrogen such as estradiol or diethylstilbestrol can induce TGF-alpha through a correctly positioned activating function 2 (AF2) and bind SRC-1. The TPE did not activate AF2 but activated the TGF-alpha gene through AF2b. This was demonstrated because D351 but not D351G ER activated the TGF-alpha gene with the TPE. We propose two classes of estrogens with different ER complexes that may incorporate different coactivators to function. Phytoestrogens and environmental xenoestrogens will fall into different classes based on structure and may exhibit selective actions and carcinogenic potential based on different ER conformations.

Binding Sites↗

Dissociation of regional activation in mild AD during visual encoding: a functional MRI study.

OBJECTIVE: The authors studied mild patients with AD with a visual learning paradigm to determine whether activations of medial temporal regions on fMRI differ in AD compared to nondemented individuals. BACKGROUND: Changes in activation patterns of medial temporal lobe regions may serve as a biologic marker of altered brain function early in the course of AD. METHODS: The authors studied eight healthy young subjects, eight late middle-age nondemented volunteers, and seven patients with mild AD. All subjects underwent fMRI scanning in which they viewed a set of geometric designs for 45 seconds. Changes in blood flow were analyzed by comparing the prestimulus fMRI signal with that present during the stimulus presentation. RESULTS: Patients with AD, who had very poor recall of the geometric designs subsequently, showed increased blood flow (activation) during stimulus presentation only in a visual association area. Both the young and older nondemented subjects, all of whom had good recall of the designs, showed activations during stimulus presentation of the right entorhinal cortex, right supramarginal gyrus, right prefrontal regions, and left anterior-inferior temporal lobe. The younger and older nondemented subjects did not differ in fMRI activation patterns. CONCLUSIONS: Failure of activation in AD of either temporal lobe or prefrontal regions is consistent with established clinical-pathologic correlations in AD. fMRI may be useful in confirming a memory disorder diagnosis and also may be useful in detecting individuals with incipient dysfunction in learning as a result of disorders such as AD.

Adult↗

Detection of GDNF secretion in glial cell culture and from transformed cell implants in the brains of live animals.

Current ex vivo gene therapy for Parkinson's disease using glial cell line-derived neurotrophic factor (GDNF) is limited by the lack of a monitoring mechanism to determine the expression of GDNF once the cells or other vehicles are transferred into animal models. The purpose of this study was to test whether a Renilla luciferase (RUC)-GDNF fusion protein secreted by the genetically engineered glial cell line RG-1 could be measured photometrically in cerebrospinal fluid (CSF). RG-1 was constructed by permanent transformation with a plasmid DNA construct that contains a GDNF cDNA (gdnf) fused to a RUC cDNA (ruc). The fusion protein secreted by RG-1 was shown to retain both GDNF and RUC activity. The concentration of GDNF determined by enzyme-linked immunoadsorbent assay (ELISA) was correlated with the light emission detected by assaying for RUC bioluminescence in RG-1 culture medium, indicating that RUC can be used as a reporter for GDNF in vitro. The cells were then implanted into rat brain (n=20), and the cisternal CSF was analyzed. Bioluminescence was successfully detected in the CSF samples, and was quantified over a period of 25 days, while Western blotting and ELISA failed to detect GDNF in CSF, presumably because the concentration of the RUC-GDNF fusion was too low. This study demonstrates that the transformed glial cell line RG-1 offers a sensitive self-reporting assay for GDNF expression.

Animals↗

Quantum mechanics simulation of protein dynamics on long timescale.

Protein structure and dynamics are the keys to a wide range of problems in biology. In principle, both can be fully understood by using quantum mechanics as the ultimate tool to unveil the molecular interactions involved. Indeed, quantum mechanics of atoms and molecules have come to play a central role in chemistry and physics. In practice, however, direct application of quantum mechanics to protein systems has been prohibited by the large molecular size of proteins. As a consequence, there is no general quantum mechanical treatment that not only exceeds the accuracy of state-of-the-art empirical models for proteins but also maintains the efficiency needed for extensive sampling in the conformational space, a requirement mandated by the complexity of protein systems. Here we show that, given recent developments in methods, a general quantum mechanical-based treatment can be constructed. We report a molecular dynamics simulation of a protein, crambin, in solution for 350 ps in which we combine a semiempirical quantum-mechanical description of the entire protein with a description of the surrounding solvent, and solvent-protein interactions based on a molecular mechanics force field. Comparison with a recent very high-resolution crystal structure of crambin (Jelsch et al., Proc Natl Acad Sci USA 2000;102:2246-2251) shows that geometrical detail is better reproduced in this simulation than when several alternate molecular mechanics force fields are used to describe the entire system of protein and solvent, even though the structure is no less flexible. Individual atomic charges deviate in both directions from "canonical" values, and some charge transfer is found between the N and C-termini. The capability of simulating protein dynamics on and beyond the few hundred ps timescale with a demonstrably accurate quantum mechanical model will bring new opportunities to extend our understanding of a range of basic processes in biology such as molecular recognition and enzyme catalysis.

Alanine↗

[Effect of dextran combination with flurosemide on nephrotic syndrome].

OBJECTIVE: To compare the diuretic effect of coadministration of dextran and flurosemide with that of coadministration of Albumin and flurosemide on primary nephrotic syndrome. METHODS: In a double-blind placebo-controlled study, eighteen primary nephrotic syndrome patients on standard sodium chloride intake, random by divided into three groups and received by intravenous administration for 60 minutes for three days (a) FU (1 mg.kg-1) combination with a sham infusion, (b) FU (1 mg.kg-1) combination with 50 ml of 20% solution of Albumin, or (c) FU (1 mg/kg) combination with 250 ml dextran 40. Urinary volume, sodium and plasma atrial nartiuretic peptide concentration were assessed. RESULTS: The results showed that administration of FU alone increased mean cumulative urinary sodium and volume excretion as compared with the administration of sham infusion without treatment (P < 0.05). The administration of FU and albumin or FU and dextran 40 caused an even more marked increase of urinary sodium and volume excretion (P < 0.01), as compared with the administration of sham infusion without treatment. Plasma ANP increased significantly on both albumin infusion days and dextran 40 infusion days (P < 0.05). CONCLUSION: Coadministration of albumin and FU or dextran 40 and FU can increase the urinary volume and urinary Na and ANP significantly, dextran 40 can take the place of albumin because there was no difference between the administration of FU and albumin and FU and dextran in UV, Una and ANP.

Adolescent↗

[Clinical X-ray analysis for 20 atypical lung cancer].

OBJECTIVE: To analyze clinical X-ray features of atypical lung cancer, expected to improve the diagnosis for atypical lung cancer. METHODS: To collect 20 cases with lung cancer from Jan. 1987 to Jan. 2000, those patients were diagnosed as pneumonia infiltration tuberculosis or lymphoma by X-ray and CT scanning firstly. To analyse retrospectly theses cases and contrast with operation and pathology. RESULTS: All patients were verified as lung cancer after following reexamine and by pathology. CONCLUSION: The cases with sigle pathy shadow in the lung fields could not exclude lung cancer when they had hemoptysis or had no typical pneumoma signs.

Adenocarcinoma↗

Homogeneous fluorescent derivatization of large proteins.

A method of homogeneously derivatizing large proteins for highly sensitive analysis is described. Homogeneity of the derivative was realized by tagging all the free amino groups of proteins. With this method, alpha-chymotrypsinogen A, ovalbumin and bovine serum albumin were derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). Prior to the derivatization, all the proteins were reduced and alkylated. After reacting the resulting unfolded proteins with excessive amounts of AQC, the samples were analyzed with matrix assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS) to determine the derivatization degree. The results indicated that all three proteins had been, or had almost been, fully derivatized. HPLC and CE were used for characterizing these protein derivatives. Under the optimized fluorescence detection conditions, the detectability of the tagged proteins was 2400-6200 times better than that detected at UV 280 nm, 170-300 times better than detected at UV 214 nm, and 150-420 times better than measured with their native fluorescence.

Electrophoresis, Capillary↗

Model-independent source imaging using two-pion correlations in (2 to 8)a GeV Au+Au collisions.

We report a particle source imaging analysis based on two-pion correlations in high multiplicity Au+Au collisions at beam energies between 2A and 8A GeV. We apply the imaging technique introduced by Brown and Danielewicz, which allows a model-independent extraction of source functions with useful accuracy out to relative pion separations of about 20 fm. The extracted source functions have Gaussian shapes. Values of source functions at zero separation are almost constant across the energy range under study. Imaging results are found to be consistent with conventional source parameters obtained from a multidimensional Hanburg-Brown-Twiss analysis.

Journal Article↗

A novel bradykinin-related peptide from skin secretions of toad Bombina maxima and its precursor containing six identical copies of the final product.

Amphibian skin contains rich bradykinin-related peptides, but the mode of biosynthesis of these peptides is unknown. In the present study, a novel bradykinin-related peptide, termed bombinakinin M, was purified from skin secretions of the Chinese red belly toad Bombina maxima. Its primary sequence was established as DLPKINRKGPRPPGFSPFR that comprises bradykinin extended from its N-terminus by a 10-residue segment DLPKINRKGP. The cDNA structure of bombinakinin M was found to contain a coding region of 624 nucleotides. The encoded precursor of bombinakinin M is composed of a signal peptide, an acidic peptide, six 100% identical copies of a 28-amino-acid peptide unit including bombinakinin M plus a spacer peptide. The sequence of bombinakinin M is preceded by a single basic residue (arginine), which represents the site of cleavage for releasing of mature bombinakinin M. This is the first cDNA cloning of bradykinin-related peptides from amphibian skin. The unique cDNA structure encoding bombinakinin M suggests that the generation modes of bradykinin-related peptides in amphibian skin and in mammalian blood system are different.

Amino Acid Sequence↗

T(11;18)(q21;q21) is associated with advanced mucosa-associated lymphoid tissue lymphoma that expresses nuclear BCL10.

The development of gastric mucosa-associated lymphoid tissue (MALT) lymphoma is a multistep process and can be clinico-pathologically divided into Helicobacter pylori-associated gastritis, low-grade tumors, and high-grade tumors. The molecular events underlying this progression are largely unknown. However, identification of the genes involved in MALT lymphoma-specific t(11;18)(q21;q21) and t(1;14)(p22;q32) has provided fresh insights into the pathogenesis of this disease. T(11;18)(q21;q21) results in a chimeric transcript between the API2 and the MALT1 genes, whereas t(1;14) (p22;q32) causes aberrant nuclear BCL10 expression. Significantly, nuclear BCL10 expression also occurs frequently in MALT lymphomas without t(1;14)(p22;q32), suggesting an important role for BCL10 in lymphoma development. Thirty-three cases of H pylori gastritis, 72 MALT lymphomas, and 11 mucosal diffuse large B-cell lymphomas (DLBCL) were screened for t(11;18)(q21;q21) by reverse transcription-polymerase chain reaction followed by sequencing. BCL10 expression in lymphoma cases was examined by immunohistochemistry. The API2--MALT1 fusion transcript was not detected in H pylori gastritis and mucosal DLBCL but was found in 25 of 72 (35%) MALT lymphomas of various sites. Nuclear BCL10 expression was seen in 28 of 53 (53%) of MALT lymphomas. Of the gastric cases, the largest group studied, the frequency of both t(11;18)(q21;q21) and nuclear BCL10 expression was significantly higher in tumors that showed dissemination to local lymph nodes or distal sites (14 of 18 = 78% and 14 of 15 = 93%, respectively) than those confined to the stomach (3 of 29 = 10% and 10 of 26 = 38%). Furthermore, t(11;18)(q21;q21) closely correlated with BCL10 nuclear expression. These results indicate that both t(11;18)(q21;q21) and BCL10 nuclear expression are associated with advanced MALT lymphoma and that their oncogenic activities may be related to each other. (Blood. 2001;98:1182-1187)

Adaptor Proteins, Signal Transducing↗

Novel boxlike dinuclear or chain polymeric silver(I) complexes with polypyridyl bridging ligands: syntheses, crystal structures, and spectroscopic and electrochemical properties.

The syntheses, characterization, crystal structures, and photophysical and electrochemical properties of two dinuclear and two polymeric Ag(I) complexes with three polypyridyl ligands, 2,3-di-2-pyridylquinoxaline (L(1)), 2,3-di-2-pyridyl-5,8-dimethoxyquinoxaline (L(2)), and 2,3,7,8-tetrakis(2-pyridyl)pyrazino[2,3-g] quinoxaline (L(3)), are described. The structures of the two boxlike dinuclear complexes with L(1) and L(2) and two chemically the same but differently crystallized one-dimensional zigzag chain coordination polymers also consisting of boxlike dinuclear subunits have been elucidated by X-ray analysis. [AgL(1)(CH(3)CN)](2)-(BF(4))(2).2CHCl(3) (1): monoclinic, C2/c; a = 28.631(2), b = 12.2259(11), c = 14.3058(12) A; beta = 99.180(2) degrees; Z = 4. [AgL(2)(CH(3)CN)(2)](2)(ClO(4))(2) (2): triclinic, P1; a = 12.3398(2), b = 13.750(2), c = 14.326(7) A; alpha = 83.494(3), beta = 74.631(3), gamma = 76.422(3) degrees; Z = 4. [[Ag(2)L(3)(NO(3))(2)].CH(3)CN](infinity) (3a): monoclinic, P2(1)/c; a = 9.5836(8), b = 13.4691(12), c = 14.0423(12) A; beta = 107.753(2) degrees; Z = 4. [Ag(2)L(3)()(NO(3))(2)](infinity) (3b): monoclinic, P2(1)/c; a = 8.4689(6), b = 16.0447(12), c = 11.7307(8) A; beta = 102.051(1) degrees; Z = 2. The structures of the dinuclear complexes 1 and 2 are similar to each other, with the two intramolecular Ag(I) centers of each complex being spanned by two ligands thus forming a unique boxlike cyclic dimer. In 1, each Ag(I) center is four-coordinated by three nitrogen atoms of two L(1) ligands and a CH(3)CN nitrogen donor, taking a distorted tetrahedral coordination geometry. The coordination environment of Ag(I) in 2 is similar to that in 1, except the formation of an additional weak coordination bond with the oxygen atom of the methoxy group of L(2). The structures of 3a,b are very similar to each other, except for the stacking patterns in the crystal lattices, and the cyclic boxlike dinuclear unit, which is similar to the structure of 1, constitutes the fundamental building block to form the one-dimensional zigzag chain structures due to the "end-on" nature of L(3). 1-3 exhibit metal-perturbed intraligand transitions in solution in 360-390 nm regions. Cyclic voltammetric studies of these complexes show the presence of reduction peak at approximately -0.5 V vs Fc(+/0). In the solid state at 77 K, they exhibit broad emission that may be assignable to originate from the metal-perturbed intraligand transitions.

Journal Article↗

Regulation of CSF1 promoter by the SWI/SNF-like BAF complex.

The mammalian BAF complex regulates gene expression by modifying chromatin structure. In this report, we identify 80 genes activated and 2 genes repressed by the BAF complex in SW-13 cells. We find that prior binding of NFI/CTF to the NFI/CTF binding site in CSF1 promoter is required for the recruitment of the BAF complex and the BAF-dependent activation of the promoter. Furthermore, the activation of the CSF1 promoter requires Z-DNA-forming sequences that are converted to Z-DNA structure upon activation by the BAF complex. The BAF complex facilitates Z-DNA formation in a nucleosomal template in vitro. We propose a model in which the BAF complex promotes Z-DNA formation which, in turn, stabilizes the open chromatin structure at the CSF1 promoter.

Base Sequence↗

Kinetics and mechanism of the beta- to alpha-CuAlCl(4) phase transition: a time-resolved (63)Cu MAS NMR and powder X-ray diffraction study.

The beta and alpha phases of CuAlCl(4) have been characterized by solid-state (27)Al and (63)Cu magic angle spinning nuclear magnetic resonance. The very short spin--lattice relaxation times of the copper spins, and the sensitivity of the I = 3/2 (63)Cu nucleus to the small differences in the local structure of Cu in the two phases, allowed (63)Cu spectra to be acquired in very short time periods (1 min), in which the beta and alpha phases were clearly resolved. This time resolution was exploited to follow the phase transition from the pseudohexagonal close-packed beta-CuAlCl(4) into the pseudocubic close-packed alpha-CuAlCl(4), which occurs above 100 degrees C. In situ time-resolved (63)Cu MAS NMR and synchrotron X-ray diffraction experiments were used to measure the kinetics of this phase transition as a function of temperature. The transformation was shown to be a first-order phase transition involving no intermediate phases with an activation energy of 138 kJ/mol. The kinetic data obey a first-order Avrami--Erofe'ev rate law. A one-dimensional growth mechanism is proposed that involves a combination of Cu(+) ion self-diffusion and a translational reorganization of the close-packed anion layers imposed by the periodic rotations of [AlCl(4)](-) tetrahedra. This beta to alpha phase transformation can be induced at ambient temperatures by low partial pressures of ethylene.

Journal Article↗