Biomedical subjects

J Hu

Publications and source records attributed to J Hu.

At least 415 records · Page 23Linked to original sources

PubMed1997

Preparation of Liposomes with a Controlled Assembly Procedure

A method of liposome preparation that involves two experimental steps is demonstrated. First, water in oil (W/O) emulsions are prepared by sonication. Then, due to centrifugal force, water droplets in the emulsion contact the monolayer at the oil/water interface and transform into liposomes after the monolayer has enwrapped the droplets. This procedure results in liposomes with diameters in the range 50-200 nm. Advantages of this procedure include relatively high (60%) encapsulation efficiency, direct preparation of liposomes in an aqueous medium different from the entrapped phase, and no leakage of the encapsulated chemicals during preparation. This relatively simple and rapid method may be useful in the encapsulation of drugs, enzymes, and other macromolecules as well as in the study of membrane properties.

Journal Article↗

PubMed1997

Differential regulation of gap junctions by proinflammatory mediators in vitro.

The development of inflammation is an important component of host defense against infection. The cellular and molecular processes underlying inflammation are well-studied, and it is known that cells of the blood vessel wall, such as endothelial cells and smooth muscle cells, play pivotal roles. Additionally, a wide variety of proinflammatory mediators have been defined, which coordinate the multicellular processes of inflammation. Knowledge of the potential role of blood vessel gap junctional intercellular communication (GJIC) in coordinating the inflammation process, however, is limited. In this study, we report that bacterial lipopolysaccharide (LPS), as well as the proinflammatory cytokines TNF-alpha and IL-1beta, selectively inhibit human myoendothelial GJIC in vitro without affecting GJIC between the respective homologous cell populations. This finding may represent a physiologically relevant component of the inflammatory response to infection. The work also provides some of the first clear evidence suggesting that a single eukaryotic cell can differentially regulate its GJIC between homologous and heterologous cell types in a simultaneous manner.

Cell Communication↗

PubMed1997

Dependence of aortic pulse wave assessments on behavioural state in normal term fetus.

To investigate any influence of behavioural states on the pulse waves in the descending aorta, 21 human fetuses were studied in utero in uncomplicated gestation at the age of 36-41 weeks. The fetal behavioural states were identified using two real-time scanners and one cardiotocograph. The aortic waveform data and pulse wave velocity (PWV) were recorded by means of two double phase-locking echo-trackers. The PWV of the fetal aorta was significantly lower in fetal behavioural state (FBS) 2F than in FBS 1F, when the fetus was apnoeic, and was also reduced in both states during fetal breathing. The calculated pulse pressure showed the same trend as the PWV. The fetal aortic end diastolic diameter and the pulse amplitude did not alter, when the two states changed. Our data suggest that the central haemodynamics in term fetuses are independently influenced by their behavioural state as well as by fetal breathing. The study of pulse waves in the fetal aorta should preferably be performed during apnoea in state 1F, when neither gross body movements nor breathing movements disturb the recording.

Aorta↗

PubMed1997

Hepadnavirus assembly and reverse transcription require a multi-component chaperone complex which is incorporated into nucleocapsids.

Assembly of hepadnaviruses depends on the formation of a ribonucleoprotein (RNP) complex comprising the viral polymerase polypeptide and an RNA segment, epsilon, present on pregenomic RNA. This interaction, in turn, activates the reverse transcription reaction, which is primed by a tyrosine residue on the polymerase. We have shown recently that the formation of this RNP complex in an avian hepadnavirus, the duck hepatitis B virus, depends on cellular factors that include the heat shock protein 90 (Hsp90). We now report that RNP formation also requires ATP hydrolysis and the function of p23, a recently identified chaperone partner for Hsp90. Furthermore, we also provide evidence that the chaperone complex is incorporated into the viral nucleocapsids in a polymerase-dependent reaction. Based on these findings, we propose a model for hepadnavirus assembly and priming of viral DNA synthesis where a dynamic, energy-driven process, mediated by a multi-component chaperone complex consisting of Hsp90, p23 and, potentially, additional factors, maintains the reverse transcriptase in a specific conformation that is competent for RNA packaging and protein priming of viral DNA synthesis.

Adenosine Triphosphate↗

PubMed1997

Combining electrospray ionization and the flowing afterglow method.

The design and implementation of a simple electrospray ionization source for a flowing afterglow/triple quadrupole device are described. Ions can be electrosprayed directly into the room temperature flow tube through a heated capillary without the need for differential pumping or ion focusing. Detected ion currents at the detector sampling orifice as high as 3 pA have been achieved, and the mass spectra indicate little or no re-clustering of the desolvated ions with the background solvent vapor in the flow tube. Spatially and temporally resolved ion/molecule reactions of electrosprayed ions can be carried out in the flow reactor under thermal energy conditions. Sufficient ion densities can be achieved for tandem mass spectrometric experiments in the triple quadrupole analyzer, including energy-resolved collision-induced dissociation. Selected chemical applications illustrating these features are described, including proton transfer reactions with aromatic polysulfonate dianions and multiply protonated polypeptides and threshold CID of a doubly charged transition metal coordination complex.

Amino Acid Sequence↗

PubMed1997

Inhibition of hepatocellular carcinoma development in hepatitis B virus transfected mice by low dietary casein.

In a comprehensive human ecological study, primary liver cancer has been shown to be highly significantly associated with 1) the prevalence of persistent infection with hepatitis B virus (HBV) and 2) plasma cholesterol concentrations that are, in turn, associated with the consumption of animal based foods. In rat studies, aflatoxin-induced hepatocellular carcinoma is substantially prevented by decreasing the intake of animal based protein (casein), a hypercholesterolemic nutrient. Thus the development of primary liver cancer associated with persistent HBV infection or with aflatoxin exposure may be controlled by reduced intake of animal-based proteins. Transgenic mice transfected with an HBV gene fragment containing the viral transactivator of hepatis B virus, HBx, which induces the formation of hepatocellular carcinoma, were used to examine the ability of dietary casein to modify tumor formation. Reducing the concentration of dietary casein to 6% from the traditional level of 22% markedly inhibited (by 75%) hepatic tumor formation in these transgenic mice. Tumor development also was substantially altered by interchanging dietary casein concentration well after tumor development had begun (at 8 months), increasing by 173% from the expected yield when casein intake was increased and decreasing by 99% when casein was reduced. These findings suggest that the development of liver tumor formation among individuals persistently infected with HBV may be controlled by minimizing or eliminating the intake of animal protein-based foods.

Animals↗

PubMed1997

Expression of the human NRAMP1 gene in professional primary phagocytes: studies in blood cells and in HL-60 promyelocytic leukemia.

In the mouse, mutations at the natural resistance-associated macrophage protein 1 (Nramp1) gene abrogate resistance to infection with antigenically unrelated intracellular parasites such as Mycobacterium, Salmonella, and Leishmania. Nramp1 expression is restricted to reticuloendothelial organs and peripheral blood leukocytes, where the protein may function as a membrane transporter of an as yet to be identified substrate. To identify the human blood cell type(s) expressing NRAMP1 mRNA and determine how Nramp1 expression is regulated in these cells, we have examined separated populations of peripheral blood leukocytes and in vitro cell lines. We observed that polymorphonuclear leukocytes (PMN) are the major site of NRAMP1 expression, followed to a lesser degree by monocytes (MN). Migration of MN to tissues (alveolar macrophages) or maturation in vitro (long-term culture) was associated with a higher level of NRAMP1 expression compared with blood MN. Northern analyses of RNA from model cultured cells showed absence of NRAMP1 expression in transformed cell lines from either erythroid or lymphoid T or B lineages as well as progenitors of the monocyte/macrophage pathway (KG1, U937, THP1), and the HL-60 promyelocytic leukemia. Induction of differentiation of HL-60 cells toward either the monocyte/macrophage (vitamin D3, phorbol ester) or the granulocyte pathways (DMF, DMSO), as measured by induction of IL8-Rb, c-FMS, and CD14 marker gene expression, was concomitant with a strong induction of NRAMP1 expression. These results suggest that NRAMP1 expression is specific to the myeloid lineage and is acquired during the maturation of PMN and MN. The possibility that NRAMP1 may be a component of the phagosomal/endosomal apparatus common to PMN and MN is discussed.

Base Sequence↗

PubMed1997

Ciliary neurotrophic factor immunoreactivity in rat intramuscular nerve during reinnervation through a silicone tube after severing of the rat sciatic nerve.

The immunoreactivity of ciliary neurotrophic factor (CNTF) and S100 was studied in the degenerating and regenerating intramuscular nerves after the sciatic nerve was severed. The sciatic nerves of male Wistar rats were transected at the midpoint of the thigh, and silicone tubing was used to obtain effective reinnervation. The strong immunoreactivity of CNTF and S100 was observed in the Schwann cell cytoplasm of intramuscular nerves (IMN) and at the neuromuscular junction (NMJ) on the control sections. The CNTF immunoreactivity gradually became weak and indistinct in the Schwann cell cytoplasm after the operation. However, it was recognized again in the IMN at 4 weeks after the operation. On the other hand, the S100 immunoreactivity was continuously observed except at the NMJ through the denervating and reinnervating period. At 12 weeks after the operation, the strong immunoreactivity of both CNTF and S100 was observed again. These findings suggest that the amount of CNTF protein decreased in Schwann cells of the IMN and NMJ during the denervating period and increased during the reinnervating period in proportion to the number of remyelinated Schwann cells after severing of the sciatic nerve. They also suggest that CNTF was more highly correlated than the S100 protein with the reinnervation activity of Schwann cells.

Animals↗

PubMed1997

Two-dimensional proton chemical-shift imaging of human muscle metabolites.

Large lipid signals and strong susceptibility gradients introduced by muscle-bone interfaces represent major technical challenges for in vivo proton MRS of human muscle. Here, the demonstration of two-dimensional proton chemical-shift imaging of human muscle metabolites is presented. This technique utilizes a chemical-shift-selective method for water and lipid suppression and automatic shimming for optimal homogeneity of the magnetic field. The 2D1H CSI technique described facilitates the acquisition of high-spatial-resolution spectra, and allows one to acquire data from multiple muscle groups in a single experiment. A preliminary investigation utilizing this technique in healthy adult males (n = 4) revealed a highly significant difference in the ratio of the creatine to trimethylamine resonance between the fast and slow twitch muscle groups examined. The technique is robust, can be implemented on a commercial scanner with relative ease, and should prove to be a useful tool for both clinical and basic investigators.

Adult↗

PubMed1997

Color Doppler monitoring of changes of utero-placental-fetal circulation in normal pregnancy and intrauterine growth retardation.

The utero-placental-fetal circulation (UPFC) of 150 subjects during second and third trimester was examined by using color Doppler. Of them 89 were normal woman and 58 were patients with intrauterine growth retardation IUGR). Our results showed that UPFC was increased gradually during normal pregnant period. In IUGR patients it was revealed that TAV and Q of UmA, UmV and UtA decreased at 20th week of gestation, especially after 30th week. PI, RI and S/D ratio of UmA were increased, but TAV, Q of UmA and UmV were markly reduced, so was UtA. PI were increased, but the changes of RI, S/D ratio in UtA were not significant. Hemodynamical findings of UmA, UmV and UtA were abnormal in 92.53% of IUGR patients. Only 81.03% present abnormal S/D ratio of UmA (P < 0.01) and the difference was statistically significant. Maternal serum E3, HPL level in IUGR were significantly lower than that of the normal. 6KP level was reduced, TXB2/6KP ratio was significantly increased. TXB2/6KP ratio was markedly related with TAV, Q of UmA, UmV and UtA. Our results suggested that using color doppler ultrasound for examination of hemodynamical changes of UmA, UmV and UtA could revealed UPFC function directly. It is one of the best methods for monitoring IUGR and might be used for early diagnosis of IUGR. The main pathophysiological changes of IUGR were UPFC obstruction and placental disfunction.

Adult↗

PubMed1997

Infection of primary human placental fibroblasts with HIV-1, HIV-2, and SIV.

Cells with fibroblast-like features were isolated from the villous tissue of normal term human placentas. Immunocytochemical characterization of the cells showed that they were vimentin-positive but negative for factor-VIII, CD14 and CD4. Thus, the cells are mesenchymal and are not endothelial cells, macrophages or trophoblast. These cells were exposed to nine different cell-free virus isolates, including seven isolates of human immunodeficiency virus type 1 (HIV-1), one HIV-2 isolate and one simian immunodeficiency virus isolate (SIVmac251). The susceptibility of the cells to infection was evaluated by immunocytochemical and virological techniques. No evidence of infection could be found using immunofluorescence microscopy or by p24 antigen capture and reverse transcriptase assays. However, virus rescue experiments using 11 different target cell types provided evidence that the placental fibroblasts were susceptible to infection with HIV-1Lai, HIV-1IIIB, HIV-2CBL-20, and SIVmac251, yet were resistant to infection by all other isolates. The infected fibroblasts exhibited neither cytopathic effects nor released virus into the culture medium. For each infected fibroblast population, some, but not all, indicator target cell lines or human peripheral blood mononuclear cells were able to rescue the respective virus. Based on these observations, we conclude that placental fibroblasts can be infected with HIV during transplacental transmission and could act as virus reservoirs, capable of infecting other fetal cells.

Animals↗

PubMed1997

Why are hepadnaviruses DNA and not RNA viruses?

Virion assembly in hepadnaviruses is a two-step process leading to (1) the packaging of viral pregenomic RNA and reverse transcriptase into nucleocapsids and (2) the assembly of nucleocapsids with envelope components, which results in the formation of mature virus particles. Characteristically, both steps are intimately coupled to viral DNA synthesis. While assembly of nucleocapsids is coupled to the protein priming of reverse transcription, virion formation is linked to genome maturation.

DNA, Viral↗

PubMed1997

Visualization of chromatin folding patterns in chicken erythrocytes by atomic force microscopy (AFM).

The organization of the higher order structure of chromatin in chicken erythrocytes has been examined with tapping-mode scanning force microscopy under conditions close to their native environment. Reproducible high-resolution AFM images of chromatin compaction at several levels can be demonstrated. An extended beads-on-astring (width of approximately 15-20 nm, height of approximately 2-3 nm for each individual nucleosome) can be consistently observed. Furthermore, superbeads (width of approximately 40 nm, height of approximately 7 nm) are demonstrated. Visualization of the solenoid conformation at the level of 30 nm chromatin fiber is attained either by using AFM or by using electron microscopy. In addition, tightly coiled chromatin fibers (approximately 50-60 nm and approximately 90-110 nm) can be revealed. Our data suggest that the chromatin in the interphase nucleus of chicken erythrocyte represents a high-order conformation and AFM provides useful high-resolution structural information concerning the folding pattern of interphase chromatin fibers.

Animals↗

PubMed1997

Adsorptive stripping voltammetry of bleomycin.

In 0.05 M H2SO4 solution, two reductive peaks, P1 and P2, of bleomycin were obtained. The peak potentials E(P1) and E(P2) were -0.83 and -1.09 V (versus Ag/AgCl), respectively. The sensitivity of P2 was much higher than that of P1. The peak current of P2 was proportional to the concentration of bleomycin over the range 1.0 x 10(-9)-1.0 x 10(-7) M with a detection limit of 5.0 x 10(-10) M using adsorptive voltammetry at an accumulation time of 120 s (Ei = -0.80 V). The behaviour of the reduction wave was studied and applied to the determination of bleomycin in mouse serum. The reduction process of P1 was irreversible with adsorptive characteristics and the adsorption behavior obeyed the Frumkin adsorptive isotherm. The adsorptive coefficient beta was 8.9 x 10(5), the interaction factor alpha was 0.94 and the Gibbs energy of adsorption delta G degree was -33.93 kJ mol-1. P2 was an irreversible adsorption peak with catalytic hydrogen properties.

Adsorption↗

PubMed1997

S100beta induces neuronal cell death through nitric oxide release from astrocytes.

The glial-derived neurotrophic protein S100beta has been implicated in the development and maintenance of the nervous system. S100beta has also been postulated to play a role in mechanisms of neuropathology because of its specific localization and selective overexpression in Alzheimer's disease. However, the exact relationship between S100beta overexpression and neurodegeneration is unclear. Recent data have demonstrated that treatment of cultured rat astrocytes with high concentrations of S100beta results in a potent activation of inducible nitric oxide synthase (iNOS) and a subsequent generation of nitric oxide (NO), which can lead to astrocytic cell death. To investigate whether S100beta-induced NO release from astroctyes might influence neurons, we studied S100beta effects on neuroblastoma B104 cells or primary hippocampal neurons co-cultured with astrocytes. We found that S100beta treatment of astrocyte-neuron co-cultures resulted in neuronal cell death by both necrosis and apoptosis. Neuronal cell death induced by S100beta required the presence of astrocytes and depended on activation of iNOS. Cell death correlated with the levels of NO and was blocked by a specific NOS inhibitor. Our data support the idea that overexpression of S100beta may be an exacerbating factor in the neurodegeneration of Alzheimer's disease.

Animals↗

PubMed1997

Increase of ciliary neurotrophic factor (CNTF) in the ischemic rat brain as determined by a sensitive enzyme-linked immunoassay.

To study the distribution of ciliary neurotrophic factor (CNTF) in the rat brain under physiological as well as pathological conditions, a sensitive sandwich enzyme linked immunoassay (EIA) for rat CNTF was developed using rabbit anti-CNTF antibody. This method detected as little as 10 pg ml-1 CNTF with a good linearity and accuracy. Analysis of various regions of rat brain and peripheral nerves showed a high content of CNTF in the sciatic nerves, spinal cord, optic nerves, and olfactory bulb. Transient focal ischemia of rat brain caused by middle cerebral artery occlusion significantly and specifically increased CNTF levels in the cerebral cortex and hippocampus regions in the ischemic hemisphere.

Animals↗

PubMed1997

Determining the amount of help used by disabled elderly persons at home: the role of coping resources.

The purpose of this study is to quantify the effects of coping resources on the amount (hours) of help used by disabled elderly persons in their home. The findings are based on the 1989 National Long-Term Care Survey. The distribution of help-hours is very skewed, mirroring the skewness of limitations in physical and cognitive functioning. Controlling for these limitations, the most important coping resources are the combinations of helpers who join forces, and coresidence with a helper. The effect of helpers' networks is large and consistent across marital status and living arrangements. The networks are more extensive for married than for unmarried persons. In reference to persons who rely only on nonrelatives: (a) a network of a spouse and children enables a married person to have 40 additional weekly hours of help, (b) a network of children and others enables an unmarried person to have 29 additional help-hours per week if he/she coresides with other adults and 10 additional weekly help-hours if he/she does not. The issue of concern for public policy is whether such family networks will be preserved and, if not, how to obtain the funds for the alternative of sufficient paid help in the community.

Adaptation, Psychological↗

PubMed1997

Identification and characterization of human genes encoding Hprp3p and Hprp4p, interacting components of the spliceosome.

Nuclear RNA splicing occurs in an RNA-protein complex, termed the spliceosome. U4/U6 snRNP is one of four essential small nuclear ribonucleoprotein (snRNP) particles (U1, U2, U5 and U4/U6) present in the spliceosome. U4/U6 snRNP contains two snRNAs (U4 and U6) and a number of proteins. We report here the identification and characterization of two human genes encoding U4/U6-associated splicing factors, Hprp3p and Hprp4p, respectively. Hprp3p is a 77 kDa protein, which is homologous to the Saccharomyces cerevisiae splicing factor Prp3p. Amino acid sequence analysis revealed two putative homologues in Caenorhabditis elegans and Schizosaccharomyces pombe. Polyclonal antibodies against Hprp3p were generated with His-tagged Hprp3p over-produced in Escherichia coli . This splicing factor can co-immunoprecipitate with U4, U6 and U5 snRNAs, suggesting that it is present in the U4/U6.U5 tri-snRNP. Hprp4p is a 58 kDa protein homologous to yeast splicing factor Prp4p. Like yeast Prp4p, the human homologue contains repeats homologous to the beta-subunit of G-proteins. These repeats are called WD repeats because there is a highly conserved dipeptide of tryptophan and aspartic acid present at the end of each repeat. The primary amino acid sequence homology between human Hprp4p and yeast Prp4p led to the discovery of two additional WD repeats in yeast Prp4p. Structural homology between these human and yeast splicing factors and the beta-subunit of G-proteins has been identified by sequence-similarity comparison and analysis of the protein folding by threading. Structural models of Hprp4p and Prp4p with a seven-blade beta-propeller topology have been generated based on the structure of beta-transducin. Hprp3p and Hprp4p have been shown to interact with each other and the first 100 amino acids of Hprp3p are not essential for this interaction. These experiments suggest that both Hprp3p and Hprp4p are components of human spliceosomes.

Amino Acid Sequence↗