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Biomedical subjects

L Tong

Publications and source records attributed to L Tong.

At least 91 records · Page 5Linked to original sources
PubMed1998

Brain-derived neurotrophic factor (BDNF) protects cultured rat cerebellar granule neurons against glucose deprivation-induced apoptosis.

In the present study, cell death induced by glucose deprivation in primary cultures of cerebellar granule neurons was examined. Glucose deprivation-induced apoptotic cell death was demonstrated using the terminal transferase-mediated (TdT) deoxyuridine triphosphate (d-UTP)-biotin nick end labeling (TUNEL) method and DNA fragmentation assays. When the effects of different neurotrophins on the survival of cerebellar granule neurons after glucose deprivation were assessed, BDNF, but not NT-3 or NGF, was found to protect cerebellar granule neurons against glucose deprivation-induced cell death. In addition, BDNF treatment increased c-Fos immunoreactivity in the cerebellar granule neurons. These results are consistent with the hypothesis that neuronal death due to glucose deprivation has a significant apoptotic component and that neurotrophins can protect against hypoglycemic damage.

Animals↗
PubMed1998

Conserved mode of peptidomimetic inhibition and substrate recognition of human cytomegalovirus protease.

Human cytomegalovirus (HCMV) protease belongs to a new class of serine proteases, with a unique polypeptide backbone fold. The crystal structure of the protease in complex with a peptidomimetic inhibitor (based on the natural substrates and covering the P4 to P1' positions) has been determined at 2.7 A resolution. The inhibitor is bound in an extended conformation, forming an anti-parallel beta-sheet with the protease. The P3 and P1 side chains are less accessible to solvent, whereas the P4 and P2 side chains are more exposed. The inhibitor binding mode shows significant similarity to those observed for peptidomimetic inhibitors or substrates of other classes of serine proteases (chymotrypsin and subtilisin). HCMV protease therefore represents example of convergent evolution. In addition, large conformational differences relative to the structure of the free enzyme are observed, which may be important for inhibitor binding.

Amino Acid Sequence↗
PubMed1998

Signal transduction in neuronal death.

Apoptosis in the nervous system is a necessary event during the development of the nervous system and is also present after genotoxic events, be they chronic as in aging or more acute after trauma and ischemia. Apoptotic events reflect an interplay between intrinsic signaling events that rely on cytokines, neurotransmitters, and growth factors and responses to extrinsic events that increase levels of radical oxygen species. Both intrinsically and extrinsically driven signal-transduction pathways act via transcription factors that regulate the coordinated timely expression of stress-response genes as part of a decision-making process that can commit cells to apoptosis or survival. Here we discuss the role of two transcription factors that participate in apoptosis in the nervous system: the activator protein AP-1 and nuclear factor kappaB.

Animals↗
PubMed1998

[Isolation and identification of triterpenoide compound from Patrinia scabiosaefolia].

In this paper, a triterpenoide was isolated from the roots and rhizomes of Patrinia scabiosaefolia Fish ex Link. Its structure was identified by combination of chemical reactions and spectrum analysis as 28-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl-hederagenin ester. The compound was found in the Patrinia for the first time.

Chromatography, Thin Layer↗
PubMed1997

Experiences from the structure determination of human cytomegalovirus protease.

Several obstacles were encountered and overcome during the structure determination of human cytomegalovirus protease. Dehydration of crystals, by exposing them to higher concentrations of the precipitant, reduced the mosaicity of the crystals and may have also resolved their microscopic twinning. The initial phase information was obtained with the selenomethionyl multiple-wavelength anomalous diffraction technique. However, site-specific mutagenesis was required to introduce extra Met residues into the protease. The phase information had to be improved by non-crystallographic symmetry averaging, initially among three 'crystal forms'. A change in the composition of the artificial mother liquor led to a significant improvement, from 3.0 and 2.0 A resolution, in the diffraction quality of the crystals. The experiences reported here may prove useful to structure determination of other proteins.

Journal Article↗
PubMed1997

Determination of Association Constants for Cyclodextrin-Surfactant Inclusion Complexes: A Numerical Method Based on Surface Tension Measurements

Inclusion complexes of beta-cyclodextrin (beta-CD) with sodium octyl sulfonate (C8As), sodium dodecyl sulfonate (C12As), and sodium hexadecyl sulfonate (C16As) in aqueous solutions are studied by surface tension measurement at the air/water interface at different temperatures. At fixed concentrations of the surfactants, the surface tension increases with an increase in beta-CD concentration to a maximum value, at which it holds. The surface tension curves of the surfactants in the presence of beta-CD are higher than those in the absence of beta-CD. The curves rise higher with the increase in beta-CD concentration for each surfactant. The apparent critical micelle concentrations (CMC*) of the surfactants vary linearly with beta-CD concentration. The CMC* and surface tension values (including those after the CMC*) for the same system decrease with increase in temperature. A numerical method based on surface tension measurements is developed to determine the association constants for 1:1 inclusion complexes. This method is very reliable and easy to perform. The results demonstrate that the longer the hydrophobic tail of the surfactant, the greater the association constant with beta-CD, and that for the same surfactant the association constant is higher at lower temperatures.

Journal Article↗
PubMed1997

Potent HIV protease inhibitors containing a novel (hydroxyethyl)amide isostere.

A series of HIV protease inhibitors containing a novel (hydroxyethyl)amidosuccinoyl core has been synthesized. These peptidomimetic structures inhibit viral protease activity at low nanomolar concentrations (IC50 < 10 nM for HIV-1 protease). The inhibition constant (Ki) for inhibitor 19 was determined to be 7.5 pM against HIV-1 and 1.2 nM against HIV-2 proteases, respectively. Several compounds (19-24) inhibited HIV-1 replication in cell culture assays with 50% effective concentrations (EC50) = 3.7-35 nM. This series of inhibitors was found to exhibit poor bioavailability (< 10%) in the rat, following oral administration. The synthesis and biological properties of these compounds are discussed. In addition, an X-ray structure of one of these inhibitors (23) in complex with HIV-2 protease provides insight into the binding mode of this novel class of HIV protease inhibitors.

Administration, Oral↗
PubMed1997

Crystallization and preliminary crystallographic analysis of recombinant human P38 MAP kinase.

The recombinant human p38 MAP kinase has been expressed and purified from both Escherichia coli and SF9 cells, and has been crystallized in two forms by the hanging drop vapor diffusion method using PEG as precipitant. Both crystal forms belong to space group P2(1)2(1)2(1). The cell parameters for crystal form 1 are a = 65.2 A, b = 74.6 A and c = 78.1 A. Those for crystal form 2 are a = 58.3 A, b = 68.3 A and c = 87.9 A. Diffraction data to 2.0 A resolution have been collected on both forms.

Animals↗
PubMed1997

Cytokine levels and inflammatory responses in developing late-phase allergic reactions in the skin.

BACKGROUND: Cytokines could play roles in the attraction of leukocytes into sites of IgE-mediated late-phase reactions (LPR) or in the activation of such cells at the sites. Previous studies found increased release of IL-6, IL-1, and granulocyte-macrophage colony-stimulating factor into skin chambers overlying allergen-challenged sites, mainly after 6 hours when LPR are already well-developed. OBJECTIVE: To compare levels of several cytokines with inflammatory responses in skin chambers overlying developing LPRs. METHODS: Skin chambers were appended to denuded blister bases in 15 sensitive subjects, then challenged over a 5-hour period with pollen allergens (Ag) or buffer control (B). Levels of several chemotactic cytokines, eosinophil cationic product (released from eosinophils), and lactoferrin (released from neutrophils) were measured and leukocyte accumulation was assessed. RESULTS: Levels of the chemokines IL-8, RANTES, and monocyte chemotactic protein-1 (but not IL-1, IL-6, or granulocyte-macrophage colony-stimulating factor) were significantly higher at Ag-challenge sites than at B-challenge sites. IL-5 was not detected. In individual chamber fluids at Ag-challenge sites, (1) IL-8 levels correlated strongly with levels of lactoferrin but not with eosinophil cationic protein levels; (2) RANTES levels correlated with numbers of eosinophils but not with eosinophil cationic protein levels; and (3) levels of monocyte chemotactic protein-1 correlated weakly with histamine released after the first hour. CONCLUSIONS: During LPRs developing during the first 5 hours of Ag challenge, there is increased local release of several chemokines, which could play roles in the observed attraction and subsequent activation of leukocytes and continued histamine release observed at such sites.

Blister↗
PubMed1997

A highly specific inhibitor of human p38 MAP kinase binds in the ATP pocket.

The crystal structure of human p38 mitogen-activated protein (MAP) kinase in complex with a potent and highly specific pyridinyl-imidazole inhibitor has been determined at 2.0 A resolution. The structure of the kinase, which is in its unphosphorylated state, is similar to that of the closely-related ERK2. The inhibitor molecule is bound in the ATP pocket. A hydrogen bond is made between the pyridyl nitrogen of the inhibitor and the main chain amido nitrogen of residue 109, analogous to the interaction from the N1 atom of ATP. The crystal structure provides possible explanations for the specificity of this class of inhibitors. Other protein kinase inhibitors may achieve their specificity through a similar mechanism. The structure also reveals a possible second binding site for this inhibitor, with currently unknown function.

Adenosine Triphosphate↗
PubMed1997

Retinoic acid induces apoptosis in PC12 cells independent of neurotrophic factors.

PC12 cells are known to undergo programmed cell death (apoptosis) when they are deprived of serum. Nerve growth factor (NGF) rescues PC12 cells from serum deprivation-induced apoptosis. In the present study, we examined the effects of retinoic acid (RA), a classic morphogen, on apoptosis in PC12 cells after serum deprivation and NGF-mediated rescue. In naive PC12 cells, all trans-RA treatment induced cell death in the presence of NGF. RA also abolished the protective effects of dibutyryl cyclic AMP or insulin under serum-free conditions. The death process was accompanied by nuclear condensation and DNA fragmentation, typical of apoptosis. In addition, RA also increased the extent of apoptosis in PC12 cells after serum deprivation. Cycloheximide, an inhibitor of protein synthesis, did not abolish the effects of RA on serum-deprived PC12 cells. RA also decreased thymidine incorporation and proliferation in NGF-treated PC12 cells. Furthermore, although the total DNA binding activity of the AP-1 transcription factor was not changed after RA treatment, RA decreased a specific AP-1 transcriptional activity. It is surprising that differentiated PC12 cells resisted the toxic effects of RA. These data suggest that RA might function as an endogenous inducer of apoptosis during neural differentiation by a mechanism distinct from that of serum deprivation.

Animals↗
PubMed1997

[Mechanisms of yiqi tongluo pill on oxygen free radicals, nitric oxide and fibrinolysis in coronary heart diseases and hypertension patients].

OBJECTIVE: To clarify the mechanism of Yiqi Tongluo Pill (YQTLP) in treating coronary heart disease and hypertension. METHODS: The clinical effects of YQTLP on 97 coronary heart diseases (CHD) and hypertension patients, in comparing to the 92 patients treated with nitroglycerin was investigated. The changes of plasma lipid peroxide (LPO), superoxide dismutase (SOD), tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) and nitric oxide (NO) were observed in the patients treated with YQTLP and nitroglycerin, which were compared to 30 healthy subjects. RESULTS: The levels of LPO and PAI increased and the levels of SOD, t-PA and NO decreased significantly in the patients. The clinical effects of YQTLP were better than that of nitroglycerin in the patients with unstable angina, while the effects of YQTLP were worse than that of nitroglycerin in the patients of hypertension and hypertension with CHD. YQTLP could decrease the levels of LPO and PAI and increase the levels of SOD, t-PA and NO significantly. The effects of YQTLP on t-PA and SOD were better than that of nitroglycerin. CONCLUSION: The protective mechanisms of YQTLP were related to inhibit lipid peroxidation, protect endothelium-derived relaxing factor and adjust the fibrinolytic activities.

Adult↗
PubMed1997

[Sudden deafness: stepwise regression analysis on the correlation factors with prognosis].

To further investigate the correlation factors of the prognosis and therapeutic efficiency of sudden deafness, 83 cases (91 ears) from 1993. 1 to 1996. 4 were studied with stepwise regression analysis. The results showed that the prognosis is correlate with the age, the hearing threshold levels of both speech and high frequencies, and the types of the audiometric curve, but not with the illness duration, sex, vertigo, ipsi or bilateral, recurrence, virus infection, cardiac or cerebral vascular diseases, and the medicine or the duration of the treatment. By comparing the results obtained from different statistical analysis, it was suggested that the unification of the criteria of the diagnosis and the therapeutic efficiency as well as the application of the multiple factors regression analysis are essential for the research on sudden deafness.

Adult↗
PubMed1996

Canine prosomatostatin: isolation of a cDNA, regulation of gene expression, and characterization of post-translational processing intermediates.

Somatostatin is a tetradecapeptide (SS-14) initially isolated from the hypothalamus that is also found in D cells of the stomach and pancreas where it exerts an inhibitory action on a variety of gastrointestinal functions. Since many of concepts important to an understanding of gastrointestinal physiology are derived from experiments in the dog we examined somatostatin gene expression and post-translational processing in the canine fundus, antrum and pancreas. The canine somatostatin cDNA which is highly homologous to other known mammalian somatostatins was used to examine somatostatin expression in isolated canine fundic D-cells. Somatostatin expression induced by cholecystokinin (10(-8) M) was inhibited by the somatostatin analog, octreotide (10(-7) M). To examine somatostatin processing in the canine gut we noted that synthesis of SS-14 and somatostatin octacosapeptide (SS-28) involves endoproteolytic cleavage of prosomatostatin (proSS) at both paired and single basic amino-acid residues, respectively. Antisera capable of recognizing the amino-terminal residues of SS-28, SS-28(1-14) and SS-28(1-12) were characterized and identified concentrations of SS-28(1-12) but not SS-28(1-14) in the fundus, antrum and pancreas equivalent to those of SS-14. Since previous biosynthetic studies in canine fundic D-cells showed that SS-14 was synthesized without the appearance of a SS-28 intermediate, we hypothesize that proSS is sequentially cleaved at a dibasic site to produce SS-14 followed by monobasic cleavage that results in the formation of SS-28(1-12). Furthermore, equivalent amounts of SS-14 and SS-28(1-12) were co-released from canine fundic D-cells by CCK (10(-8) M) suggesting that the generation of these products occurs within the same regulated pathway of secretion.

Amino Acid Sequence↗
PubMed1996

A new serine-protease fold revealed by the crystal structure of human cytomegalovirus protease.

Human cytomegalovirus (hCMV), a herpesvirus, infects up to 70% of the general population in the United States and can cause morbidity and mortality in immunosuppressed individuals (organ-transplant recipients and AIDS patients) and congenitally infected newborns. hCMV protease is essential for the production of mature infectious virions, as it performs proteolytic processing near the carboxy terminus (M-site) of the viral assembly protein precursor. hCMV protease is a serine protease, although it has little homology to other clans of serine proteases. Here we report the crystal structure of hCMV protease at 2.0 angstroms resolution, and show that it possesses a new polypeptide backbone fold. Ser 132 and His 63 are found in close proximity in the active site, confirming earlier biochemical and mutagenesis studies. The structure suggests that the third member of the triad is probably His 157. A dimer of the protease with an extensive interface is found in the crystal structure. This structure information will help in the design and optimization of inhibitors against herpesvirus proteases.

Crystallography, X-Ray↗