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H Zhu

Publications and source records attributed to H Zhu.

At least 415 records · Page 23Linked to original sources
PubMed1996

RAGE and amyloid-beta peptide neurotoxicity in Alzheimer's disease.

Amyloid-beta peptide is central to the pathology of Alzheimer's disease, because it is neurotoxic--directly by inducing oxidant stress, and indirectly by activating microglia. A specific cell-surface acceptor site that could focus its effects on target cells has been postulated but not identified. Here we present evidence that the 'receptor for advanced glycation end products' (RAGE) is such a receptor, and that it mediates effects of the peptide on neurons and microglia. Increased expressing of RAGE in Alzheimer's disease brain indicates that it is relevant to the pathogenesis of neuronal dysfunction and death.

Alzheimer Disease↗
PubMed1996

Physiologically based kinetic model of effector cell biodistribution in mammals: implications for adoptive immunotherapy.

The goal of the present investigation was to develop a physiologically based kinetic model to describe the biodistribution of immunologically active effector cells in normal and neoplastic tissues of mammals based on the current understanding of lymphocyte trafficking pathways and signals. The model was used to extrapolate biodistribution among different animal species and to identify differences among different effector populations and between intra-arterial and systemic injections. Most importantly, the model was used to discern critical parameters for improving the delivery of effector cells. In the model, the mammalian body was divided into 12 organ compartments, interconnected in anatomic fashion. Each compartment was characterized by blood flow rate, organ volume and lymphatic flow rate, and other physiological and immunological parameters. The resulting set of 45 differential equations was solved numerically. The model was used to simulate the following biodistribution data: (a) nonactivated T lymphocytes in rats; (b) interleukin 2-activated tumor-infiltrating lymphocytes in humans; (c) nonactivated natural killer (NK) cells in rats; and (d) interleukin 2-activated adherent NK cells in mice. Comparisons between simulations and data demonstrated the feasibility of the model and the scaling scheme. The similarities as well as differences in biodistribution of different lymphocyte populations were revealed as results of their trafficking properties. The importance of lymphocyte infiltration from surrounding normal tissues into tumor tissue was found to depend on lymphocyte migration rate, tumor size, and host organ. The study confirmed that treatment with effector cells has not been as impressive as originally promised, due, in part, to the biodistribution problems. The model simulations demonstrated that low effector concentrations in the systemic circulation greatly limited their delivery to tumor. This was due to high retention in normal tissues, especially in the lung. Reducing normal tissue retention through decreasing attachment rate or adhesion site density in the lung by 50% could increase the tumor uptake by approximately 40% for tumor-infiltrating lymphocytes and by approximately 60% for adherent NK cells. Our analysis suggested the following strategies to improve effector cell delivery to tumor: (a) bypassing the initial lung entrapment with administration to the arterial supply of tumor; (b) reducing normal tissue retention using effector cells with high deformability or blocking lymphocyte adhesion to normal vessels; and (c) enhancing tumor-specific capture and arrest by modifying the tumor microenvironment.

Animals↗
PubMed1996

In vitro T lymphopoiesis of human and rhesus CD34+ progenitor cells.

Differentiation of hematopoietic progenitor cells into T lymphocytes generally occurs in the unique environment of the thymus, a feature that has hindered efforts to model this process in the laboratory. We now report that thymic stromal cultures from rhesus macaques can support T-cell differentiation of human or rhesus CD34+ progenitor cells. Culture of rhesus or human CD34+ bone marrow-derived cells depleted of CD34+ lymphocytes on rhesus thymic stromal monolayers yielded CD3+CD4+CD8+, CD3+CD4+CD8-, and CD3+CD4-CD8+ cells after 10 to 14 days. In addition to classical T lymphocytes, a discrete population of CD3+CD8loCD16+CD56+ cells was detected after 14 days in cultures inoculated with rhesus CD34+ cells. CD3+ T cells arising from these cultures were not derived from contaminating T cells present in the CD34+ cells used to inoculate thymic stromal monolayers or from the thymic monolayers, as shown by labeling of cells with the lipophilic membrane dye PKH26. Expression of the recombinase activation gene RAG-2, which is selectively expressed in developing lymphocytes, was detectable in thymic cultures inoculated with CD34+ cells but not in CD34+ cells before thymic culture or in thymic stromal monolayers alone. Reverse transcriptase-polymerase chain reaction analysis of T cells derived from thymic stromal cultures of rhesus and human CD34+ cells showed a polyclonal T-cell receptor repertoire. T-cell progeny derived from rhesus CD34+ cells cultured on thymic stroma supported vigorous simian immunodeficiency virus replication in the absence of exogenous mitogenic stimuli. Rhesus thymic stromal cultures provide a convenient means to analyze T-cell differentiation in vitro and may be useful as a model of hematopoietic stem cell therapy for diseases of T cells, including acquired immunodeficiency syndrome.

Animals↗
PubMed1996

Benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethylketone (Z-VAD.FMK) inhibits apoptosis by blocking the processing of CPP32.

Interleukin-1 beta converting enzyme (ICE)-like proteases, which are synthesized as inactive precursors, play a key role in the induction of apoptosis. We now demonstrate that benzyloxycarbonyl-Val-Ala-Asp (OMe) fluoromethylketone (Z-VAD.FMK), an ICE-like protease inhibitor, inhibits apoptosis by preventing the processing of CPP32 to its active form. These results suggest that novel inhibitors of apoptosis can be developed which prevent processing of proforms of ICE-like proteases.

Amino Acid Chloromethyl Ketones↗
PubMed1996

Autocrine growth loop of the epidermal growth factor receptor in normal and immortalized human bronchial epithelial cells.

Epidermal growth factor (EGF) is a potent growth factor for human normal bronchial epithelial (HBE) cells and lung cancer cells, which often demonstrate an EGF receptor (EGFR) autocrine loop. We have found that HBE cells are capable of proliferating in basal medium without EGF supplementation, and this suggests the probable presence of an active EGFR autocrine loop in non-neoplastic HBE cells. Northern blot hybridization shows that the parental and immortalized HBE cells express comparable and high levels of mRNA for EGFR, transforming growth factor-alpha (TGF-alpha), and amphiregulin (AR), but not EGF. Incubation with neutralizing monoclonal antibodies (mAb) against EGFR partially inhibits the growth of these cells. Immunohistochemistry shows that HBE cells express the TGF-alpha peptide in vitro and in vivo, however, neutralizing mAbs against TGF-alpha fail to inhibit their proliferation. In contrast, AR stimulates the growth of HBE cells. Thus, several EGF-family ligands appear to be involved functionally in the EGFR autocrine growth loop in HBE cells.

Amphiregulin↗
PubMed1996

Liposome-encapsulated dichloromethylene diphosphonate induces macrophage apoptosis in vivo and in vitro.

Dichloromethylene diphosphonate (MDPCl2) encapsulated in multilamellar liposomes was selectively incorporated by macrophages, immediately transferred to lysosomes, then released from liposomes into lysosomes by enzymatic digestion of the liposomal lipid layers. From 4 h after ingesting liposome-encapsulated MDPCL2 murine macrophages in vivo and in vitro acquired the ultrastructural features of apoptosis, such as condensed nuclear chromatic, nuclear fragmentation, cell shrinkage, and blebbing of the plasma membrane. Murine peritoneal macrophages and isolated rat Kupffer cells incubated in the medium containing liposome-encapsulated MDPCl2 increased DNA fragmentation in a dose-dependent manner. Electrophoretic analysis of extracted DNA from the isolated Kupffer cells showed DNA fragmentation. Another diphosphonate, Alendronate (4-amino-1-hydroxy-butylidene-1,1-diphosphonate) had less potent macrophage cytotoxicity. However, MDPCl2, Alendronate, and gadolinium chloride in solution were not cytotoxic to macrophages. These results implied that the intralysosomal accumulation of MDPCl2 generates signals to induce macrophage apoptosis.

Alendronate↗
PubMed1996

Mu- and delta-opioid receptor antagonists precipitate similar withdrawal phenomena in butorphanol and morphine dependence.

The relative involvement of mu- and delta-opioid receptors in the mediation of butorphanol-, as compared to morphine-, dependence was examined with the use of highly selective antagonists at mu- and delta-opioid receptors. Extracellular fluid levels of glutamate (Glu) and aspartate (Asp) were measured within the pontine locus coeruleus following precipitation of withdrawal from dependence on either butorphanol or morphine in conscious Sprague-Dawley rats. Dependence was induced by intracerebroventricular (i.c.v.) infusion of butorphanol (26 nmol/mu l/h), morphine (26 nmol/mu l/h) or saline vehicle (1 mu l/h) for 3 days by means of an osmotic minipump. Microdialysis probes (2 mm tip) were inserted into the locus coeruleus 24 h before precipitation of withdrawal by i.c.v. injection of either the mu-opioid receptor antagonist, D-Pen-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP; 48 nmol/5 mu l or 48 nmol/5 mu l), or the delta-opioid receptor antagonist, naltrindole (17-cyclopropy;methyl-6,7-dehydro-4,5-epoxy-3, 14-dihydroxy-6,7,2'3'-indolmorphinan hydrochloride; 48 nmol/5 mu l or 100 nmol/5 mu l). Baseline levels of Glu ranged from 9.59 + or - 1.27 to 12.84 + or - 3.01 mu M in the various treatment groups. Levels of Asp were similar. Precipitation of withdrawal by CTOP elicited significant increases of Glu and Asp in both morphine- and butorphanol-dependent rats. Maximal increases in Glu of 425% and 258% above baseline levels were elicited in the first 15 min microdialysis sample following i.c.v. injection of CTOP in morphine- and butorphanol-dependent rats, respectively. Behavioral signs of withdrawal were greater in morphine than butorphanol-dependent groups. The i.c.v. treatment with naltrindole elicited increases in Glu and Asp that were similar, although less marked, than those precipitated by CTOP treatment. Administration of naltrindole produced equivalent signs of withdrawal in both morphine- and butorphanol-dependent rats. Withdrawal from dependence on both morphine and butorphanol is characterized by elevations in coerulear levels of excitatory amino acids. Responses elicited following the use of selective mu- and delta-opioid receptor antagonists to precipitate withdrawal suggest that the role played by these receptors in mediation of the signs and symptoms of withdrawal do not differ greatly between butorphanol- and morphine-dependent rats.

Animals↗
PubMed1996

Effects of glutathione and pH on the oxidation of biomarkers of cellular oxidative stress.

Cellular oxidative stress is associated with such pathological conditions as arteriosclerosis, inflammatory diseases and cancer. The oxidation of the biomarkers. 2',7'-dichlorofluorescin (DCFH), 2-deoxyribose, and lipid peroxidation are often used to assess the status of oxidative stress in cells and tissues. Since high levels of reduced glutathione (GSH) and acidic conditions have been associated with diminished chemical lethality, we evaluated the influence of these parameters on the cellular response to oxidative stress. We used a cultured hepatocyte line (ch/ch cells) that is susceptible to oxidative toxicity. A hydroxyl radical-generating system consisting of H2O2, ascorbate and iron produced a pH-dependent lethality, with complete cell killing at pH 7.4 and none at pH 6.8. Lethality correlated with the depletion of intracellular GSH, and with an increase in DNA fragmentation. The influence of GSH and pH was assessed for DCFH and 2-deoxyribose oxidation, and for lipid peroxidation. The oxidation of DCFH and 2-deoxyribose was inhibited by GSH, with about 4-fold greater inhibition efficacy at pH 6.8 than at pH 7.4 [IC50 values (microM GSH) for pH 6.8 and 7.4, respectively: DCFH = 7 and 30; 2-deoxyribose = 125 and 490]. GSH did not affect lipid peroxidation at either pH, even at a high intracellular concentration of 10 mM. We conclude: 1) GSH is not inhibiting DCFH and 2-deoxyribose oxidation by simply quenching reactive oxygen (hydroxyl radical or perferryl oxygen), since GSH did not inhibit lipid peroxidation: 2) the protonated form GSH is more likely to be the inhibitory species rather than GS-, since even in the simple cell-free systems lower pH inhibited biomarker oxidation; and; 3) hydroxyl radical may not be the primary intracellular oxidant of DCFH, since intracellular GSH concentrations are typically 10- to 100-fold higher than the IC50 values for GSH inhibiting reactive oxygen-mediated DCFH oxidation.

Animals↗
PubMed1996

Physicochemical characterization of nedocromil bivalent metal salt hydrates. 1. Nedocromil magnesium.

Nedocromil sodium is used in the treatment of reversible obstructive airways diseases, such as asthma. The physicochemical, mechanical, and biological characteristics of nedocromil sodium can be altered by its conversion to other salt forms. In this study, three crystalline hydrates, the pentahydrate, heptahydrate, and decahydrate, of a bivalent metal salt, nedocromil magnesium (NM), were prepared. The relationships between these hydrates were studied through their characterization by differential scanning calorimetry (DSC), thermogravimetric analysis (TGA). Karl Fischer titrimetry (KFT), hot stage microscopy (HSM), ambient or variable temperature powder X-ray diffraction (PXRD), Fourier-transform infrared (FTIR) spectroscopy, solid-state nuclear magnetic resonance (SSNMR) spectroscopy, scanning electron microscopy (SEM), water uptake at various relative humidities (RH), intrinsic dissolution rate (IDR), and solubility measurements. The pentahydrate showed two dehydration steps, corresponding to two binding states of water, a more temperature-sensitive tetramer and a more stable monomer, deduced from the crystal structure previously determined. The heptahydrate and decahydrate each showed a dehydration step with a minor change in slope at about 50 degrees C, which was analyzed by derivative TGA and confirmed by DSC. HSM and variable temperature PXRD also confirmed the thermal dehydration behavior of the NM hydrates. The decahydrate underwent an apparently irreversible phase transformation to the pentahydrate at 75 degrees C at an elevated water vapor pressure. The PXRD, FTIR, and SSNMR of the decahydrate were similar to those of the heptahydrate, suggesting that the three extra water molecules in the decahydrate are loosely bound, but were significantly different from those of the pentahydrate. The rank order of both IDR and solubility in water at 25 degrees C was heptahydrate approximately decahydrate > pentahydrate, corresponding to the rank order of free energy with respect to the aqueous solution.

Differential Thermal Analysis↗
PubMed1996

Generation of human T lymphocytes from bone marrow CD34+ cells in vitro.

Analysis of the events that regulate development of red blood cells or granulocytes has led to therapies altering clinical conditions associated with anemia or neutropenia. The development of therapeutic approaches to target conditions associated with lymphopenia, such as AIDS, has been thwarted by limited techniques for studying T-lymphocyte development. We describe an in vitro system in which human bone marrow CD34+ cells proliferate, acquire the expression of the lymphoid-specific RAG-2 gene and a broad repertoire of rearranged T-cell receptor genes, develop the ability to produce T cell-specific interleukin-2 and achieve a range of T-cell immunophenotypes. The cells also become susceptible to infection with the T-lymphotropic strain of human immunodeficiency virus-1, HIV-1IIIB. This culture system induces human T lymphopoiesis and may permit further analysis of the events regulating human T-lineage differentiation. It provides a preclinical model for screening stem cell gene therapies directed toward AIDS.

Adult↗
PubMed1996

Phosphorus-31 nuclear magnetic resonance spectroscopy of the spinal cord in the pig, rat, and rabbit.

RATIONALE AND OBJECTIVES: To ensure that contamination-free phosphorus-31 nuclear magnetic resonance (31P-NMR) spectra of the spinal cord could be obtained, a porcine model was adopted that provided a large cord sample and a greater area free from adjacent muscle tissue. METHODS: Phosphorus-31 NMR spectra were acquired from the porcine spinal cord under in vivo conditions using a 4.7-T spectrometer. Spectra also were collected from perchloric acid and lipid extracts, and excised freeze trapped samples of the rat, rabbit, and pig spinal cord. RESULTS: The in vivo spectrum showed resonances corresponding to adenosine triphosphate, phosphocreatine, inorganic phosphate, phosphomonoesterase, and phosphodiesterase as confirmed by extracts. In addition, a broad resonance was observed that was assigned to myelin phospholipids. CONCLUSION: Phosphorus-31 NMR spectra of the spinal cord revealed resonances common to brain tissue. Importantly, the existence of a previously undetected resonance, which is likely to correspond to myelin phospholipids, also is reported. This resonance may prove important in future studies monitoring changes in myelin in response to trauma and ischemia.

Animals↗
PubMed1996

An optoelectric plantar "shear" sensing transducer: design, validation, and preliminary subject tests.

A prototype miniature plantar shear sensing transducer was developed, characterized, and tested in this study. Electro-optical components were chosen for the design because of the fast response time, low cost, small size, low power requirements, and adaptability to this application. The optoelectric circuit employed a 660 nm wavelength light source and photodiode solar cell. Signal amplification and sensitivity were adjusted to provide an output voltage proportional to light power. The sensor shell was designed to encapsulate the electro-optical sensing components while providing mechanical resistance to shear through a spring mechanism. A naval bronze was chosen for the shell due to its strength and nonreflective characteristics (alloy of copper and tin). Static and dynamic characteristics of the shear sensor were determined through a series of calibration tests. Mechanical crosstalk sensitivity ranged from 14.34 to 30.51 mV/N. This represented 1% full-scale/Newton sensitivity. Nonlinearity averaged 5.6% in the forward direction and 7.6% in the reverse direction. Overall sensor output hysteresis was 1.1 +/- 3.1% while the natural frequency of the sensor to an input shear transient was approximately 5 Hz. Temperature sensitivity was -7.0 mV/degree C or 3.5% full-scale/degree C. Testing of five adult subjects revealed peak anterior-posterior shear ranging from 6.7 kPa (posterior heel) to 51.4 kPa (great toe) and medial-lateral shear ranging from 5.4 kPa (great toe) to 43.5 kPa (first metatarsal head). Stress-time integral values ranged from 0.78 kPa-sec (posterior shear at the posterior heel) to 37.3 kPa-sec (medial shear at the posterior heel). Contact durations ranged from 0.28 sec (posterior shear at the posterior heel) to 1.25 sec (medial shear at the posterior heel). Further application of the sensor for plantar shear characterization in able-bodied subjects and those with pathology is suggested.

Adult↗
PubMed1996

Spontaneous melanocytosis in transgenic mice.

Clone B is a 2-kb fragment of cloned genomic DNA involved in adipocyte differentiation in vitro. Insertion of this DNA fragment into the genome of a variety of cell lines results in committing the recipient cells to undergo adipocyte differentiation. Construction of transgenic mice with Clone B DNA resulted in an unexpected phenotype--spontaneous melanocytosis. The present study describes the distribution and morphology of melanin-containing lesions in these transgenic mice. Spontaneous melanin-containing dermal lesions appeared on the ears, snout, and perianal regions of transgenic mice by the age of 3-4 months. Multifocal dermal masses rapidly developed into raised lesions, which appeared to spread to adjacent skin. Ultrastructural examination of lymph nodes, spleen, and dermal lesions of these mice revealed membrane-bound melanin with effacement f the organelle structure of severely affected cells. Protein gel electrophoresis revealed elevated activity of tyrosinase in the pinnae, skin, perianal mass, and lymph nodes. This line of transgenic mice may provide a useful model for investigation of the etiology and progression of benign and malignant melanin-containing tumors.

Animals↗
PubMed1996

Insulin-like growth factor II induces DNA synthesis in fetal ventricular myocytes in vitro.

Insulin-like growth factor II (IGF2) belongs to a family of growth factors that includes insulin and insulin-like growth factor I (IGF1). Although the accumulating evidence indicates that IGF1 is involved in regulating proliferation of ventricular myocytes, the role of IGF2 is less clear. To gain more insight into the functions of IGF2, rat ventricular expression of IGF2 mRNA at four developmental stages was examined by Northern analysis. An abundant IGF2 mRNA of approximately 3.8 kb was detected in fetal ventricles. It was dramatically decreased in neonatal ventricles and became undetectable in juvenile and adult ventricles. Similar expression patterns of the mRNA encoding IGF1 receptor and IGF2 receptor were observed. Since the results of Northern analysis strongly suggest the importance of IGF2 in regulating proliferation of fetal rat ventricular myocytes, the effects of an exogenous IGF2 on DNA synthesis in cultured rat ventricular myocytes were determined. DNA synthesis, which was monitored by measuring 5-bromo-2'-deoxyuridine (BrdU) and [3H]thymidine incorporation, was increased by twofold to threefold in IGF2-stimulated fetal ventricular myocytes, whereas no change in BrdU or [3H]thymidine incorporation was observed in neonatal ventricular myocytes. Instead, IGF2 seemed to induce hypertrophy in neonatal ventricular myocytes. An antisense oligonucleotide against rat IGF2 mRNA was able to significantly reduce BrdU incorporation, and this effect was quantitatively reversed by the addition of exogenous IGF2. Reversion by exogenous IGF2 was abolished by a monoclonal antibody against IGF1 receptor. In conclusion, our results suggest that IGF2 directly regulates proliferation of fetal rat ventricular myocytes in a paracrine/autocrine fashion.

Animals↗
PubMed1996

Analysis of target cell susceptibility as a basis for the development of a chemoprotective strategy against benzene-induced hematotoxicities.

A goal of our research is to identify biochemical factors that underlie the susceptibility of bone marrow cell populations to benzene metabolites so as to develop a mechanistically based chemoprotective strategy that may be used in susceptible humans exposed to benzene. By doing biochemical risk analysis of bone marrow stromal cells from mice and rats and the human myeloid cell lines, HL-60 and ML-1; and by using buthionine sulfoximine and dicumarol we have observed that the susceptibility of these cell populations to hydroquinone (HQ) correlates with their concentration of glutathione (GSH) and activity of quinone reductase (QR). Accordingly, the induction of QR and GSH by 1,2-dithiole-3-thione (D3T) in these cell populations has resulted in a significant protection against the following hydroquinone-mediated toxicities: inhibition of cell proliferation and viability; reduced ability of stromal cells to support myelopoiesis; and altered differentiated of ML-1 cells to monocytes/macrophages. Preliminary in vivo experiments indicate that feeding mice D3T results in an induction of QR in the bone marrow compartment such that stromal cells are more resistant to hydroquinone-induced cytotoxicity in vitro. Overall, these studies suggest that in addition to hepatic cytochrome P4502E1, bone marrow QR and GSH are factors that could determine an individual's relative susceptibility to the toxic effects of benzene.

Animals↗
PubMed1996

Four isolates of Giardia lamblia cultivated axenically in China and the restriction endonuclease analysis of their DNA.

Axenic culture of 4 Giardia lamblia isolates, 3 from humans and 1 from a rabbit, have been established in China for the first time. The trophozoites of Giardia used for the initial cultivation were obtained from the purified cysts excysted in suckling gerbil (Meriones unguiculatus) and BALB/c mice. Modified TYI-S-33 medium was used in establishment of the culture. DNA banding patterns were compared using restriction-endonuclease analysis and then followed by Southern blot analysis with genomic DNA as a probe. The 4 isolates showed identical banding patterns after being digested with Hind III, Bgl II, Pts I, Hae III, and Hinf. Two different banding patterns were observed after further digestion with Alu I and southern blot analysis. This indicated that isolates of human and rabbit origins do not have strict host specificity and that cross transmission may occur between humans and rabbits.

Adolescent↗