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Biomedical subjects

H Liu

Publications and source records attributed to H Liu.

At least 1,261 records · Page 70Linked to original sources
PubMedDate not supplied

Characterization of absorption and scattering properties for various yeast strains by time-resolved spectroscopy.

An understanding of the optical properties of biological media and cells is essential to the development of noninvasive optical studies of tissues. Unicellular organisms offer a unique opportunity to investigate the factors affecting light propagation, since they can be manipulated in ways impossible for more complex biological samples. In this study, we examined optical absorption and scattering properties of strongly multiple scattering yeast suspensions by means of near-infrared (NIR) time-resolved spectroscopy (TRS) and a sample substitution method. We determined the critical parameters for photon migration by varying the cell organelle content, the cell ploidy, the cell size, and the concentration of suspended cells. The results indicate that the photon absorption is insensitive to cell differentiation and that the cell volume is the primary factor determining light-scattering property.

Cell Count↗
PubMedDate not supplied

Molecular cloning and sequence analysis of cDNAs encoding a beta-toxin-like peptide and two MkTx I homologues from scorpion Buthus martensii Karsch.

Three full-length cDNAs, one encoding the precursor of a beta-toxin-like peptide (named BmKBT) and the other two encoding those of (MkTx I) homologues (named MkTx II and MkTx III, respectively), were isolated from a venom gland cDNA library of the Chinese scorpion Buthus martensii Karsch, by screening with a cDNA fragment generated by PCR. The encoded precursor of BmKBT contained 83 amino acid residues including a signal peptide of 19 residues, a mature peptide of 63 residues and an extra basic residue (Lys) which have to be removed in the processing step. The deduced amino acid sequence of BmKBT showed 52% homology to that of beta-neurotoxin TsVII isolated from scorpion Tityus serrulatus. However, the positions of disulfide bridges have a little variation between the two peptides. The precursors of MkTx II and MkTx III both contained 85 amino acid residues including a signal peptide of 19 residues, a mature peptide of 64 residues and two extra residues (Gly-Arg) which have to be removed in the processing step, too. There was high sequence similarity (90%) between the two peptides. The sequences of mature MkTx II and MkTx III were highly homologous with MkTx I isolated from scorpion Buthus martensii Karsch, both showing 90% identities.

Amino Acid Sequence↗
PubMedDate not supplied

Molecular cloning and genomic organization of a K(+) channel toxin from the Chinese scorpion Buthus martensii Karsch.

A full-length cDNA encoding the precursor of a K(+) channel toxin (BmTX2) was first isolated from a venom-gland cDNA library of the Chinese scorpion Buthus martensii Karsch. The precursor is composed of a signal peptide of 21 residues and a mature toxin of 37 residues with three disulfide bridges. The genomic gene of BmTX2 was also cloned and sequenced. It consisted of two exons, disrupted by an intron of 81 bp inserted in the region encoding signal peptide.

Amino Acid Sequence↗
PubMedDate not supplied

Structural features and regulatory properties of the brain glutamate decarboxylases.

It is widely recognized that the two major forms of GAD present in adult vertebrate brains are each composed of two major sequence domains that differ in size and degree of similarity. The amino-terminal domain is smaller and shows little sequence identity between the two forms. This domain is thought to mediate the subcellular targeting of the two GADs. Substantial parts of the amino-terminal domain appear to be exposed and flexible, as shown by proteolysis experiments and the locations of posttranslational modifications. The carboxyl-terminal sequence domain contains the catalytic site and shows substantial sequence similarity between the forms. The interaction of GAD with its cofactor, pyridoxal-5' phosphate (pyridoxal-P), plays a key role in the regulation of GAD activity. Although GAD(65) and GAD(67) interact differently with pyridoxal-P, their cofactor-binding sites contain the same set of nine putative cofactor-binding residues and have the same basic structural fold. Thus the cofactor-binding differences cannot be attributed to fundamental structural differences between the GADs but must result from subtle modifications of the basic cofactor-binding fold. The presence of another conserved motif suggests that the carboxyl-terminal domain is composed of two functional domains: the cofactor-binding domain and a small domain that closes when the substrate binds. Finally, GAD is a dimeric enzyme and conserved features of GADs superfamily of pyridoxal-P proteins indicate the dimer-forming interactions are mediated mainly by the carboxyl-terminal domain.

Animals↗
PubMedDate not supplied

Specifically targeting the CD22 receptor of human B-cell lymphomas with RNA damaging agents.

Targeting CD22 on human B-cells with a monoclonal antibody conjugated to a cytotoxic RNAse causes potent and specific killing of the lymphoma cells in vitro. This translates to anti-tumor effects in human lymphoma models in SCID mice. RNA damage caused by RNAses could be an important alternative to standard DNA damaging chemotherapeutics. Moreover, targeted RNAses may overcome problems of toxicity and immunogenicity associated with plant or bacterial toxin containing immunotoxins.

Animals↗
PubMedDate not supplied

Anchor-chain molecular system for orientation control in enzyme immobilization.

An anchor-chain molecular system was constructed for controlled orientation and high activity in enzyme immobilization. A streptavidin recognition peptide (streptag) coding sequence was fused to the 3' end of the phoA gene, which codes for E. coli alkaline phosphatase (EAP). Both the wild-type (WT) and the Asp-101 --> Ser (D1O1S) mutant were modified with the streptag sequence with or without the insertion of a flexible linker peptide [-(Gly-Ser)(5)-] coding sequence. The fused genes were cloned into the vector pASK75 and expressed in the periplasm of the host cell Escherichia coli SM547. The proteins were released by osmotic shock and purified by ion-exchange chromatography. Enzyme activities of all proteins were measured spectrophotometrically with rho-nitrophenyl phosphate as the substrate. Specific activities of D101S-streptag and D101S-linker-streptag enzymes were increased 25- or 34-fold over the WT, respectively. These fusion proteins were then immobilized on microtiter plates through streptag-streptavidin binding reaction. After immobilization, the D101S-linker-streptag enzyme displayed the highest residual activity and the ratio of enzyme activities of the linker to nonlinker enzymes was 8.4. These results show that the addition of a linker peptide provides a spacer so as to minimize steric hindrance between the enzyme and streptavidin. The method provides a solution for controlled enzyme immobilization with high recover activity, which is especially important in construction of biosensors, biochips, or other biodevices.

Amino Acid Sequence↗
PubMedDate not supplied

Synthesis and terminal functionalization of a polymerizable phosphatidylethanolamine.

We report the design and synthesis of bifunctional phospholipid conjugates, which contain a polymerizable acrylate group and a terminal linker, such as biotin or N-(epsilon-maleimidocaproyl (EMC), to facilitate bioconjugation reactions. The lipid conjugate can be used to generate a multifunctional substrate-supported phospholipid film that is further stabilized via in-situ photocopolymerization.

Acrylates↗
PubMedDate not supplied

Construction of a fusion enzyme system by gene splicing as a new molecular recognition element for a sequence biosensor.

A bifunctional fusion enzyme system constructed by gene splicing is proposed as a new model to develop sequence biosensors, taking maltose biosensor as an example. The cDNA fragment of Aspergillus niger glucoamylase (E.C 3.2.1.3, GA) was fused to the 3' end of Aspergillus niger glucose oxidase (E.C 1.1.3.4, GOD) gene with the insertion of a flexible linker peptide [-(Ser-Gly)5-] coding sequence. The fusion gene was cloned into the vector pPIC9 and expressed in Pichia pastoris GS115 under the control of the AOX1 promoter. It was found that a bifunctional hybrid protein with a molecular weight of 430 kDa was secreted after induction with methanol. The fusion enzyme GOD-(Ser-Gly)5-GA (GLG) was purified using Q Sepharose Fast Flow ion-exchange chromatography. Kinetic analysis demonstrated that GLG retained the typical kinetic properties of both GA and GOD. After being immobilized on an aminosilanized glass slide through covalent bonding by glutaraldehyde, GLG showed much higher sequential catalytic efficiency than the mixture of separately expressed GA and GOD (GA/GOD). Maltose biosensors were fabricated with GLG and GA/GOD, respectively. The performance characteristics of the maltose biosensor with respect to reproducibility, signal level, and linearity were effectively improved by using the fusion enzyme. Our findings offer a basis for the development of other sequence biosensors.

Artificial Gene Fusion↗
PubMedDate not supplied

Affinity extraction of BSA with reversed micellar system composed of unbound Cibacron Blue.

Affinity Cibacron Blue 3GA (CB) dye in aqueous phase was directly transferred to the reversed micelles due to electrostatic interaction between anionic CB and cationic cetyltrimethylammonium bromide (CTAB). The bovine serum albumin (BSA) transfer to the reverse micelles increases significantly in a wide range of pH by the addition of a small amount of CB ( approximately 1.0-7.0% of the total surfactant concentration) to the aqueous phase. For pH < pI, the selectivity can be significantly improved with the presence of affinity CB because no BSA was extracted in the absence of CB. For backward extraction of BSA from the micellar phase with stripping aqueous solution, the addition of 2-propanol to the aqueous phase can recover almost all BSA (98.5%) extracted into the reverse micelles.

Animals↗
PubMedDate not supplied

Effect of ligand structure on formation and DNA binding properties of the transformed rat cytosolic aryl hydrocarbon receptor.

The saturation binding of transformed rat hepatic aryl hydrocarbon (Ah) receptor with [32P]-dioxin responsive element was determined using gel mobility shift assays. The assay was carried out with ligands which exhibit both high and low Ah receptor agonist activity, namely: (high) 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), 1,2,3,7,8-pentachlorodibenzo-p-dioxin, and 1,2,3,7,8-pentachlorodibenzofuran; and (low) 1,2,7,8-TCDF, 2,3,7-trichlorodibenzo-p-dioxin and 6-methyl-1,3,8-trichlorodibenzofuran (MCDF). Woolfplot analysis of the saturation binding data revealed that for the concentrations of ligands used in this study the Bmax values for the high-affinity ligands were significantly higher (36.9-39.4 fmol/mg) than observed for the low-affinity ligands (6.15-13.8 fmol/mg). In contrast, there was not a structure-dependent trend in the equilibrium constant for dissociation KD values or in the half-lives (t1/2) for decomposition of the transformed receptor complexes. MCDF, a partial Ah receptor agonist/antagonist, also transformed the cytosolic Ah receptor and inhibited TCDD-induced transformation as determined by gel mobility shift assays. However, the diagnostic value of this assay to detect partial Ah receptor antagonists is questionable since similar results were obtained with 1,2,7,8-TCDF, a ligand which does not exhibit partial antagonist activity.

Animals↗
PubMedDate not supplied

Very low birth weight births at non-NICU hospitals: the role of sociodemographic, perinatal, and geographic factors.

PURPOSE: The purpose of this study was to assess the extent of variation in the percentage of very low birth weight (VLBW) infants born at perinatal Level 1 hospitals (no Neonatal Intensive Care Unit [NICU]) across California's nine geographic Perinatal regions. The role of sociodemographic, perinatal, and geographic factors was also assessed. METHODS: Multivariate analysis of California birth certificate files between 1989 and 1993, for 24,094 live-born infants weighing between 500 and 1499 gm, was conducted to identify factors associated with delivery at a Level 1 hospital. Analyses specific for race and ethnicity were also conducted for Hispanic, African American, and white cohorts. RESULTS: In the 5-year study period, 1989 through 1993, 10.5% (24,094) of all live-born VLBW infants were delivered in Level 1 hospitals. Significant variation across regions was evident, ranging from a regional low of 3.1% to a high of 24.3%. After controlling for multiple factors, the odds of delivering at a Level 1 hospital were decreased for African Americans and South East Asians and increased in Hispanic women as compared with white non-Hispanic women. For all women, less then adequate prenatal care, living in a 50% to 75% urban zip code, and living greater then 25 miles from the nearest NICU significantly increased the odds of VLBW delivery at a Level 1 hospital. For Hispanics, teen pregnancy and having two or more prior infant deaths increased the odds, whereas Medi-Cal as the payer source for delivery and two or more pregnancy complications decreased the odds of a Level 1 VLBW delivery. After taking these factors into account, when compared with Los Angeles, the odds of inappropriate delivery site ranged from 0.37 to 2.75 across California's nine geographic perinatal regions. Of this variation, 78% could be accounted for by the percentage of total births that delivered at a region's Level 1 hospitals. CONCLUSION: The overall state average of 10.5% deliveries of VLBW at Level 1 hospitals, although close to the 10% national objective for the year 2000, did not indicate the wide variation seen across California's nine geographic regions. Risk-adjusted regional differences in the likelihood of inappropriate delivery site for the high-risk VLBW infants suggest that reaching the Healthy People 2000 objective will require further strengthening of California's perinatal regional networks, especially in those regions where a high percentage of total births deliver at Level 1 hospitals.

California↗
PubMedDate not supplied

Sensitivity and specificity of human T-lymphotropic virus (HTLV) types I and II polymerase chain reaction and several serologic assays in screening a population with a high prevalence of HTLV-II.

BACKGROUND: Since 1988, all blood donations in the United States have been screened for antibodies to human T-lymphotropic virus type I (HTLV-I). However, the sensitivity of current serologic tests for the detection of HTLV type II (HTLV-II) antibodies and the diagnostic utility of direct tests for HTLV-I and -II using polymerase chain reaction (PCR) are poorly defined. STUDY DESIGN AND METHODS: Five hundred sixty-nine HTLV-I- or -II-seropositive and 687 age- and sex-matched seronegative samples from a high-risk population at an inner-city emergency department were selected. All samples were tested with four HTLV enzyme immunoassays (EIAs), one Western blot assay and one type-specific Western blot assay, one HTLV type-specific EIA, and a research HTLV-I/II PCR kit. RESULTS: Sensitivity of the various EIAs ranged from 95.1 to 99.5 percent, and specificity ranged from 97.2 to 99.4 percent. PCR performed in duplicate without selective retesting had lower sensitivity (85.1 %) and specificity (88.0%). However, PCR detected 20 (3.2%) HTLV-I-positive and 47 (7.5%) HTLV-II-positive samples among the 627 samples that were negative in all EIAs. The type-specific EIA and PCR assay had the highest rate of concordance in classifying samples as either HTLV-I or II, with the type-specific EIA and type-specific Western blot having the next highest rates of concordance. CONCLUSION: In this sample set from a population at high risk for HTLV-II, screening with HTLV-I/II PCR had lower sensitivity and specificity than that with EIAs. However, 4.1 to 10.8 percent of samples were PCR positive but seronegative for HTLV-I or -II, and their true infection status remains undetermined.

Adult↗
PubMedDate not supplied

Comparison of fetal lung maturation in preterm singleton and twin pregnancies.

Preterm labor or premature rupture of membranes is one of the major complications of twin pregnancies and contributes to the increased maternal and perinatal morbidity and mortality. Forty-seven twin pregnancies and a comparable group of singleton pregnancies constitute the study population. These patients' pregnancies were complicated only by preterm labor. The lecithin to sphingomyelin ratio is used as an index of fetal lung maturation. Analysis of the data reveals that fetal lung maturation is biochemically comparable between twin and singleton pregnancies and among twins for a given gestational age in this clinical setting.

Adult↗
PubMedDate not supplied

MR-guided and MR-monitored neurosurgical procedures at 1.5 T.

A combined MR suite and operating room (MR-OR) has been developed and extensively assessed for its use in a wide spectrum of therapeutic applications. Equipped with a 1.5 T short bore clinical MR scanner and standard neurosurgical OR equipment, in this MR surgical suite, surgeons can obtain intraoperative planar and volumetric MR images with superior soft tissue contrast and spatial resolution for surgical planning, guidance, and monitoring. Besides MR morphologic imaging capability, blood oxygen level-dependent functional MRI and proton MR spectroscopic imaging have been demonstrated intraoperatively in the same MR-OR to aid in surgical planning and guide tumor resections. A perspective surgical navigation device and remotely operated instrument have been developed and successfully used to assist surgeons in aligning and introducing biopsy needles under fluoroscopic MRI in brain biopsy procedures. Furthermore, surgical complications can be assessed immediately before the closure. There are numerous advantages offered by this unprecedented MR-guided surgical approach, most of which are demonstrated and presented herein. Since 1997, >270 neurosurgical cases (42% brain biopsies, 25% tumor resections, 11% functional neurosurgeries, 10% cyst drainages and shunt placements, and 12% others) have been performed in the MR-OR with a <1% overall complication rate. The tumor recurrence rate for the MR-guided surgical approach is significantly less than that of the conventional one. Exemplary neurosurgical cases that have been performed in the MR-OR suite within the last 24 months are included. Overall, this high magnetic field approach to the MR-guided minimally invasive surgical procedures has been shown to be practical and acceptable to neurosurgeons as well as to neuroradiologists for a wide range of neurosurgical and neuroradiologic applications.

Biopsy, Needle↗
PubMedDate not supplied

A comprehensive study of the relation between serum concentrations, concentration ratios, and level/dose ratios of carbamazepine and its metabolites with age, weight, dose, and clearances in epileptic children.

We made a comprehensive study of the relation between age, weight, carbamazepine (CBZ) dose, total clearance (TC), and intrinsic clearance (IC) and concentrations, concentration ratios, and level/dose ratios of CBZ, carbamazepine-10,11-epoxide (CBZ-E) and trans-10,11-dihydroxy-10,11- dihydro-carbamazepine (CBZ-H) in a group of epileptic children receiving CBZ monotherapy. Body weight and age showed negative correlations with TC, IC, CBZ dose, and CBZ-E/CBZ and CBZ-H/CBZ concentration ratios, and had positive relation with CBZ, CBZ-E, and CBZ-H level/dose ratios. These results indicate decreased CBZ metabolism with patient maturity. Correlations between CBZ dose with TC, IC, and the concentration ratios of CBZ-E/CBZ, CBZ-H/CBZ-E, and CBZ-H/CBZ were positive. CBZ dose also had negative associations with CBZ and CBZ-E level/dose ratios, indicating dose-dependent autoinduction of CBZ metabolism. Our data suggest that weight, age, and CBZ dose have less influence on epoxide-hydrolase activities than on epoxidase activities. The CBZ-E/CBZ concentration ratio can be used as an indicator of the degree of autoinduction of CBZ metabolism, even in patients receiving CBZ monotherapy.

Adolescent↗
PubMedDate not supplied

Charge-coupled device detector: performance considerations and potential for small-field mammographic imaging applications.

The physical characteristics of a charge-coupled device (CCD) image detector were evaluated, as well as its potential as a digital imaging device for small field mammographic applications such as preoperative needle localization. The detection system is based on a 2048 x 2048 pixel CCD operated in 1024 x 1024 mode. The CCD was optically coupled to an intensifying screen via a lens, without intermediate intensification. The thermal noise was suppressed to 0.15 electrons pixel-1s-1 by cooling the CCD with liquid nitrogen. The dominant source of noise was attributed to the on-chip amplifier during the readout process that was performed at 50,000 pixels s-1. The measured readout noise level was 15 electrons per pixel. The low-noise characteristics of this CCD prototype detector produced encouraging results under conditions simulating mammography, with a signal level close to one electron per pixel for each detected x ray. The mean glandular dose to the breast, based on the entrance exposure measured from a standard mammographic phantom would be 1.52 mGy (152 mrad). The ultimate spatial resolution of the system was approximately 8 cycles/mm but it was limited to about 5 cycles/mm when operated in the 1024 x 1024 imaging mode. Other physical characteristics of the system such as optical coupling efficiency, exposure response, and signal-to-noise ratio were evaluated. The results of this study suggest that the use of a scientific-grade CCD allows for very good low-contrast discrimination and moderate spatial resolution under conditions simulating mammography, but the current prototype is limited to a 9 x 9-cm2 field of view.(ABSTRACT TRUNCATED AT 250 WORDS)

Female↗