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Biomedical subjects

N Zhou

Publications and source records attributed to N Zhou.

At least 19 recordsLinked to original sources
PubMed1998

Three-dimensional solution structure of lactoferricin B, an antimicrobial peptide derived from bovine lactoferrin.

The solution structure of bovine lactoferricin (LfcinB) has been determined using 2D 1H NMR spectroscopy. LfcinB is a 25-residue antimicrobial peptide released by pepsin cleavage of lactoferrin, an 80 kDa iron-binding glycoprotein with many immunologically important functions. The NMR structure of LfcinB reveals a somewhat distorted antiparallel beta-sheet. This contrasts with the X-ray structure of bovine lactoferrin, in which residues 1-13 (of LfcinB) form an alpha-helix. Hence, this region of lactoferricin B appears able to adopt a helical or sheetlike conformation, similar to what has been proposed for the amyloidogenic prion proteins and Alzheimer's beta-peptides. LfcinB has an extended hydrophobic surface comprised of residues Phe1, Cys3, Trp6, Trp8, Pro16, Ile18, and Cys20. The side chains of these residues are well-defined in the NMR structure. Many hydrophilic and positively charged residues surround the hydrophobic surface, giving LfcinB an amphipathic character. LfcinB bears numerous similarities to a vast number of cationic peptides which exert their antimicrobial activities through membrane disruption. The structures of many of these peptides have been well characterized, and models of their membrane-permeabilizing mechanisms have been proposed. The NMR solution structure of LfcinB may be more relevant to membrane interaction than that suggested by the X-ray structure of intact lactoferrin. Based on the solution structure, it is now possible to propose potential mechanisms for the antimicrobial action of LfcinB.

Amino Acid Sequence↗
PubMed1997

Cloning and expression in Escherichia coli of a human gelatinase B-inhibitory single-chain immunoglobulin variable fragment (scFv).

The murine monoclonal antibody REGA-3G12 selectively and specifically inhibits the activity of human gelatinase B. The cDNA fragments which encode the variable regions of the light and heavy chains were isolated by PCR-mediated cloning and sequenced. Single-chain Fv expression constructs for Escherichia coli were generated in which c-myc tag sequences were encoded. Inducible expression of the scFv and secretion to the periplasm were obtained with higher yields when the c-myc tag sequence was positioned at the amino-terminal side. The inhibitory activity of purified scFv on neutrophil gelatinase B was tested in a gelatin degradation assay and it was found to possess a similar specific activity as that of the intact monoclonal antibody and of the pepsin-clipped F(ab')2 derivative. This shows for the first time that inhibition of soluble enzymes with scFv is possible and opens new perspectives for the treatment of diseases with excessive and detrimental enzyme production in the host.

Amino Acid Sequence↗
PubMed1997

Does oral contraceptive use increase the risk of breast cancer in women with BRCA1/BRCA2 mutations more than in other women?

We conducted a study to determine whether the risk of breast cancer associated with oral contraceptive (OC) use is higher in women with BRCA1/BRCA2 mutations than in other women by examining whether breast cancer patients with these mutations were more likely than breast cancer patients without mutations in BRCA1/BRCA2 to have used OCs. We tested for BRCA1 185delAG and 5382insC and BRCA2 6174delT mutations in a population-based sample of 50 young Ashkenazi Jewish breast cancer patients. Nine patients (18%) had a BRCA1 mutation, and five patients (10%) had a BRCA2 mutation. Long-term OC use (>48 months) before a first full-term pregnancy was associated with an elevated risk of being classified as a mutBRCA carrier (odds ratio, 7.8; trend, P = 0.004). The results suggest that OC use may increase the risk of breast cancer more in mutBRCA carriers than in noncarriers; however, they must be interpreted with caution given the small sample size.

Adult↗
PubMed1997

Class I MHC mediates programmed cell death in human lymphoid cells.

BACKGROUND: Although signaling via class I MHC molecules has been shown to suppress T-cell responses, the mechanisms by which these effects are mediated have not been delineated. Studies were conducted to examine the possibility that 5H7 (a murine anti-human mAb specific for the alpha3 domain of human class I MHC) induces programmed cell death (PCD). MATERIALS: Normal human T cells and lymphocyte tumor lines were used. PCD was assessed by viable cell recovery (VCR) with trypan blue, ethidium bromide/acridine orange staining, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling techniques. RESULTS: 5H7 induced growth inhibition of lymphocyte tumor cell lines as assessed by [3H]thymidine incorporation. 5H7 also induced marked reductions in VCR of lymphocyte tumors and normal human T and B cells, with the most dramatic reductions occurring in B cells and B cell-derived tumors (JY, BeVD, and 521). Reductions in tumor growth observed with 5H7 monoclonal antibody, however, were not observed with other anti-human class I MHC monoclonal antibodies, TP2599 (alpha3 domain specific) and W6/32 (alpha2/alpha3 domain specific). Cells treated with 5H7 demonstrated typical features of PCD including cytoplasmic vacuolization, DNA condensation, and apoptotic body formation. Reduction in VCR was augmented by anti-CD3 monoclonal antibody and was not reversed by exogenous interleukin 2. Induction of PCD was not observed with soluble 5H7 F(ab)'2 fragments alone, but cross-linking of 5H7 F(ab)'2 fragments by F(ab)'2 fragments of human Ig-absorbed goat anti-mouse Ig partially restored PCD induction, indicating that anti-class I MHC monoclonal antibody can induce PCD in an FcR-independent system and that monoclonal antibody cross-linking is necessary for PCD induction. CONCLUSIONS: These studies provide the first evidence that class I MHC molecules mediate PCD in human T and B cells.

Antibodies, Monoclonal↗
PubMed1997

NMR studies of caldesmon-calmodulin interactions.

The binding of the calcium-regulatory protein calmodulin (CaM) to caldesmon (CaD) contributes to the regulation of smooth muscle contraction. Two regions of caldesmon have been identified as putative calmodulin-binding domains. We have earlier reported on the binding of one of these domains to calmodulin (Zhang & Vogel (1994) Biochemistry 33, 1163-1171). Here we have studied the binding of CaM to synthetic peptides of CaD which contain: (1) both the first and second CaM-binding domains; (2) the second CaM-binding domain; and (3) the sequence between the first and second CaM-binding domains. Two-dimensional transferred nuclear Overhauser enhancement proton NMR measurements as well as circular dichroism studies of a 22-residue peptide NKETAGLKVGVSSRINEWLTK, which contains the second CaM-binding domain, show that only the C-terminal half of the peptide becomes alpha-helical upon binding to CaM. Somewhat surprisingly, the shorter 9-residue peptide SRINEWLTK was sufficient to form a 1:1 complex with CaM; this peptide appears to bind as a 3(10)-helix. Proton-carbon-13 correlation NMR titration studies with specifically labeled [methyl-13C]methionine CaM were used to study the participation of the hydrophobic regions in both domains of the dumbbell shaped CaM in peptide binding. Binding of a 54-residue CaD peptide containing both CaM-binding domains affects all the 8 Met residues in the two hydrophobic domains of CaM (only Met 76 in the linker region of CaM is not involved), while binding of the second CaM-binding domain of CaD influences principally Met 51, 71, and Met 124, 144. Simultaneous binding to CaM of two peptides comprising the first and the second CaM-binding domains also caused changes to all Met residues except Met 76. Taken together, these data demonstrate that both CaM-binding domains of CaD can bind simultaneously to the two hydrophobic regions of CaM.

Amino Acid Sequence↗
PubMed1997

Nitinol alloy endotracheal stent for treatment of tracheal stenosis.

OBJECTIVE: To treat inoperable tracheal stenosis by using nitinol endotracheal stent and to solve some problems facing endotracheal stenting with other kinds of stents. METHODS: The stent was made of nitinol wire. It can be deformed to smaller size in iced water. With a self-made introducer, the stent was placed in the location of stenosis. After hot water was injected into the introducer, the stent resumed its original form and anchored there itself. RESULTS: Altogether, 5 patients with severe tracheal stenosis received this treatment. Dyspnea was relieved immediately after implantation of stents in all patients. Long-term follow-up also showed satisfactory results. CONCLUSION: Nitinol stent has some remarkable advantages as compared with other tracheal stents. It can be used as a new endoprosthesis in treating narrowed trachea.

Adult↗
PubMed1996

The nuclear hormone receptor coactivator SRC-1 is a specific target of p300.

p300 and its family member, CREB-binding protein (CBP), function as key transcriptional coactivators by virtue of their interaction with the activated forms of certain transcription factors. In a search for additional cellular targets of p300/CBP, a protein-protein cloning strategy, surprisingly identified SRC-1, a coactivator involved in nuclear hormone receptor transcriptional activity, as a p300/CBP interactive protein. p300 and SRC-1 interact, specifically, in vitro and they also form complexes in vivo. Moreover, we show that SRC-1 encodes a new member of the basic helix-loop-helix-PAS domain family and that it physically interacts with the retinoic acid receptor in response to hormone binding. Together, these results implicate p300 as a component of the retinoic acid signaling pathway, operating, in part, through specific interaction with a nuclear hormone receptor coactivator, SRC-1.

Amino Acid Sequence↗
PubMed1996

Characterization of a novel influenza hemagglutinin, H15: criteria for determination of influenza A subtypes.

Two viruses with a novel hemagglutinin (HA), A/duck/Australia/341/83 and A/shearwater/West Australia/2576/79, have been isolated from a duck and a shorebird in Australia. Hemagglutination inhibition and double immunodiffusion assays failed to reveal cross-reactivity with any of the known subtypes (H1 to H14). We therefore propose that these viruses constitute a new HA subtype, H15. Sequence analysis of the HA genes confirmed the serologic findings. When compared at the amino acid level, the HA1 region of the H15 subtype differs from those of the other subtypes by 30% and more. This degree of heterogeneity is also found among HA genes of other subtypes. Thus we propose that amino acid sequence data should be evaluated when determining the HA subtypes of influenza A viruses. Sequence comparison and phylogenetic analysis suggested that the HA subtype H15 is most closely related to the H7. Compared to the H7 HA, the H15 acquired a 30-nucleotide insertion within HA1 at position 253 which is located in the globular head of the molecule. This finding suggests that RNA recombination, although a rare event in nature, may play an important role in the evolution of influenza viruses.

Amino Acid Sequence↗
PubMed1996

Airborne emissions at skin surfaces: a potential biological exposure index.

Dermal exposures of methanol were administered in a clinical study designed to compare several biological indicators. Four subjects were exposed in five exposure sessions of varying length. In each session, a sequence of measurements of methanol concentrations in blood, breath, and headspace samples of air at exposed and unexposed skin were collected before and after dermal exposures. Skin headspace samples, collected in gas sampling bags, were designed to reflect equilibrium skin: air partitioning. At exposed skin, headspace samples were highly elevated for at least 8 h following exposure, indicating the presence of a methanol reservoir in skin. After exposure, methanol concentrations at exposed skin showed a rapid initial decline, then a slower first-order decrease. Methanol concentrations were clearly detectable in headspace samples at unexposed skin. Substantial transfer from exposed skin occurred due to mechanical contact and washing. When transfer was restricted, surface concentrations at unexposed skin were similar to levels in breath and were strongly correlated to methanol concentrations in blood. While results are preliminary due to the small sample sizes and several unresolved experimental issues, the simple, rapid, and noninvasive skin headspace measurements appear useful as a biological exposure indicator that clearly shows the presence and site of a dermal exposure, and measurements at unexposed skin reflect concentrations in blood.

Adult↗
PubMed1996

Influenza infection in humans and pigs in southeastern China.

The three last pandemic strains of influenza A virus-Asian/57, Hong Kong/68 and Russian/77-are believed to have originated in China. The strains responsible for the 1957 and 1968 human pandemics were reassortants incorporating both human and avian influenza viruses, which may have arisen in pigs. We therefore undertook a population-based study in the Nanchang region of Central China to establish the prevalence, types and seasonal pattern of human influenza infection and to screen serum samples from animals and humans for evidence of interspecies transmission of influenza viruses. Two definite influenza seasons were demonstrated, one extending from November to March and the other July to September. The profile of antibodies to commonly circulating human influenza viruses was no different in Nanchang and neighboring rural communities than in Memphis, Tennessee, USA. In particular, Chinese women who raised pigs in their homes were no more likely to have been exposed to influenza virus than were subjects who seldom or never had contact with pigs. However, we did obtain evidence using isolated H7 protein in an enzyme-linked immunoabsorbent assay for infection of pig farmers by an avian H7 influenza virus suggesting that influenza. A viruses may have been transmitted directly from ducks to humans. The results of the serological survey also indicated that pigs in or near Nanchang were infected by human H1N1 and H3N2 influenza viruses, but not with typical swine viruses. We found no serological evidence for H2 influenza viruses in humans after 1968.

Adolescent↗
PubMed1995

Cobalt-permeable non-NMDA receptors in developing chick brainstem auditory nuclei.

Kainate, an agonist at non-NMDA glutamate receptors, evoked strong concentration-dependent cobalt accumulation in the chick brain stem auditory nuclei angularis, laminaris (NL) and magnocellularis (NM). This effect could be blocked completely by the AMPA/kainate receptor antagonist CNQX and the AMPA antagonist GYKI 53655 but not by antagonists of NMDA receptors or voltage-sensitive calcium channels. Kainate (30 microM) evoked cobalt uptake from embryonic day (E)9 through E21, with a peak intensity at E15. Before E13, uptake occurred mainly in NL but declined markedly in NM after E15 and ceased in NL by E21. We conclude that calcium-permeable non-NMDA receptors are transiently expressed at the time when mature neuronal number, form and synaptic connectivity are established.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1995

High expression of HBV S gene in Bombyx mori cell culture and in silkworms.

The coding sequence of human hepatitis B virus (HBV) subtype adr surface antigen S gene was inserted into the genome of the nuclear polyhedrosis virus of Bombyx mori (BmNPV) under control of the polyhedrin promoter, and a recombinant BmNPVS virus was constructed. Upon infection of the recombinant virus, the yield of HBsAg was 35.5 micrograms per 10(6) cells, 750 micrograms per larva, and 690 micrograms per pupa. Western blot and electron microscope observation of the preliminary purified HBsAg particles showed that they were well glycosylated and assembled as regular 22-nm (diameter) particles. The CsCl buoyant density was 1.2 g/mL, which is consistent with that of HBsAg in the blood of patients.

Animals↗
PubMed1993

Maintenance of pharmacologically-immature glutamate receptors by aberrant synapses in the chick cochlear nucleus.

Surgical destruction of the otocyst in chick embryos prevents formation of the *** ear, abolishes normal cochlear input to the cochlear nucleus (nucleus magnocellularis, NM) and results in axons from the contralateral NM forming (in addition to their normal bilateral endings in nucleus laminaris, NL) a novel and functional aberrant projection to the deafferented NM. We studied the pharmacology of synaptic transmission at aberrant synapses in an in vitro preparation of the brainstem in chick embryos and hatchlings. Transmission at the aberrant synapses (as with cochlear nerve synapses in NM and NM synapses in NL) is blocked by the quinoxalinedione antagonists CNQX and NBQX, confirming the presence of excitatory amino acid receptors of the non-NMDA subtype. At cochlear nerve synapses in NM, the antagonist potency of NBQX normally decreases rapidly after embryonic day (E)18 (IC50 = 0.69 +/- 0.06 microM, mean +/- S.E.M.), reaching an asymptotic value by E21 (IC50 = 2.7 +/- 0.4 microM) that is maintained at least through posthatching day (P)14 (IC50 = 3.6 +/- 0.3 microM). In the case of the aberrant endings, the potency of NBQX remained (from E21 [IC50 = 0.6 +/- 0.1 microM] through at least P14[IC50 = 0.5 +/- 0.1 microM]) at levels that are statistically indistinguishable from the E18 value for normal cochlear nerve synapses.(ABSTRACT TRUNCATED AT 250 WORDS)

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1993

Two-dimensional NMR and restrained molecular dynamics studies of the hairpin d(T8C4A8): detection of an extraloop cytosine.

The 1H and 31P NMR resonances of the partly self-complementary 20-mer DNA d(T8C4A8) were assigned by two-dimensional HOHAHA, NOESY, and heteronuclear COSY NMR spectroscopy. The chemical shifts, NOEs, and H-H coupling patterns are indicative of the formation of a hairpin structure with the four C residues forming a loop and the T8.A8 portion of a double-stranded stem. The observation of unusual across-strand NOEs between the A H2 and the T H1' of the corresponding 3'-end neighboring base pairs of the stem residues suggests that the structure of the hairpin stem deviates from regular B-DNA. A total number of 296 interproton NOEs were used as approximate proton-proton distance constraints in restrained molecular dynamics calculations. Several different starting models, all generated manually from standard B-DNA coordinates, gave rise to virtually the same refined hairpin structure. In the final structure, the interior A-T base pairs of the hairpin stem show a high degree of propeller twist as well as base pair buckle, while the minor groove is slightly narrower compared with a normal B-DNA structure; these features are all common to bent DNA. The first three A-T pairs from the end of the hairpin have a propeller twist and base pair buckle which more closely resemble those of regular B-DNA. The four-residue loop was formed mainly by variations in the phosphate backbone torsion angle epsilon at the loop-stem junctions (residues 8 and 13) and at the first C residue (C 9). The base of the first C residue is positioned outside of the loop.(ABSTRACT TRUNCATED AT 250 WORDS)

DNA↗
PubMed1993

Solution structure of the parallel-stranded hairpin d(T8 C4A8) as determined by two-dimensional NMR.

The structure of the oligodeoxynucleotide (3')T8(5')-(5')C4A8(3') hairpin in aqueous solution was studied by two-dimensional (2D) proton and phosphorus nuclear magnetic resonance (NMR) spectroscopy. At 2.5 mM and 10 degrees C, the molecule exists predominantly as a monomolecular hairpin with a C4 loop. At higher concentrations and lower temperatures, NMR signals from multimers are obvious. They account for approximately 25% of the total population at 4 mM and 10 degrees C. Nearly all of the proton NMR signals for the hairpin could be assigned using 2D COSY, HOHAHA, and NOESY experiments. 2D 1H-31P correlation experiments were used to assign all the phosphorus resonances and to provide an additional check for the sequential assignments. A parallel-stranded T8.A8 stem can be formed in the hairpin due to the presence of the unusual 5'-5' linkage in the loop. 2D NOESY experiments indicate that the A H2 and its 5'-end neighbor base pair T methyl protons are within 5 A of each other. This is in accord with reverse Watson-Crick base pairing between T and A, which locates the A H2 and the T methyl protons in the same groove of the duplex. The chemical shifts of A H1', H2', and H2" sugar and the H2 base protons are quite different compared to normal B-DNA. Analysis of the 2D COSY and NOESY cross peak patterns indicates that the deoxyribose rings are mainly in the C2'-endo conformation and that the stem forms a right-handed helix, with the two strands held together by eight reverse Watson-Crick A.T base pairs to form a parallel-stranded duplex. The backbone torsion angles, as determined from the 31P chemical shifts, are slightly different for the A and the T residues. A molecular model was constructed, using a total of 336 proton NOE cross peak intensities as proton-proton distance constraints. In the refined structure, the conformations of the sugar-phosphate linkage, the deoxyribose rings, and the glycosyl bonds for the two parallel strands of the hairpin are close to a regular B-DNA structure. The base-stacking and the hydrogen-bonding interactions are well optimized; however, the two grooves are of approximately equal width. Thus, compared to B-DNA, the parallel-stranded duplex has a very different surface shape, and because of the reverse Watson-Crick base pairing, it has different groups exposed in each groove.

DNA↗
PubMed1993

Gamma-D-glutamylaminomethyl sulfonic acid (GAMS) distinguishes kainic acid- from AMPA-induced responses in Xenopus oocytes expressing chick brain glutamate receptors.

The effects of the glutamate receptor antagonist gamma-D-glutamylaminomethyl sulfonic acid (GAMS) on inward currents induced by bath application of kainic acid (KA) or alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) were studied with single-electrode voltage clamp methods in Xenopus oocytes injected 3-5 days previously with mRNA from the brain of E16-17 chick embryos. Both AMPA and KA induced smooth inward currents, with Hill coefficients of 1.5 (AMPA) and 2.1 (KA). GAMS, at concentrations up to 1 mM, produced no reliable antagonism of AMPA-induced currents but showed a consistent, dose-dependent and reversible antagonism of KA-induced responses; the slope of the Schild plot was 0.76 and the pA2 value 4.32. In the presence of GAMS, however, the Hill coefficient for AMPA is reduced significantly and approaches unity, suggesting that AMPA interacts with both KA and AMPA binding sites on chick brain glutamate receptors. The selectivities of three quinoxalinedione antagonists (6,7-dinitroquinoxaline-2,3-dione [DNQX], 6-cyano-7- nitroquinoxaline-2,3-dione [CNQX] and 6-nitro-7-sulfamoyl-benzo(F)quinoxaline-2,3-dione [NBQX]) were then compared with that shown by GAMS. DNQX, CNQX and NBQX all blocked the effects of both KA and AMPA completely, competitively, reversibly and dose-dependently, with Schild-plot slopes very close to 1.0. Against AMPA, observed pA2 values were 6.58 for DNQX, 6.43 for CNQX and 6.77 for NBQX. Against KA, pA2 values were 6.42 for DNQX, 6.56 for CNQX and 7.21 for NBQX.(ABSTRACT TRUNCATED AT 250 WORDS)

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1992

Developmental changes in the effects of drugs acting at NMDA or non-NMDA receptors on synaptic transmission in the chick cochlear nucleus (nuc. magnocellularis).

The developmental pharmacology of excitatory amino acid (EAA) receptors in the chick cochlear nucleus (nucleus magnocellularis, NM) was studied by means of bath application of drugs and recording of synaptically-evoked field potentials in brain slices taken from chicks aged embryonic day (E) 14 through hatching (E21). The abilities of various EAA agonists (N-methyl-D-aspartate [NMDA], kainic acid, and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid [AMPA]) to suppress postsynaptic responses by depolarization block and of EAA antagonists ((3-[RS]-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid [CCP], dizocilpine [MK-801], 6-nitro-7-sulfamoyl-benzo(F)quinoxaline-2,3 dione [NBQX], 6-cyano-7-nitroquinoxaline-2,3-dione [CNQX] and 6,7-dinitroquinoxaline-2,3-dione [DNQX]) to suppress these responses directly were assessed quantitatively. The results support the existence of NMDA receptors in NM and suggest that the ability of these receptors to influence synaptically-evoked responses declines dramatically during the last week of embryonic life. The results similarly suggest that the non-NMDA receptors in NM undergo changes in density and/or function during a period of development when the cochlear nucleus is undergoing a variety of morphological and functional transformations.

6-Cyano-7-nitroquinoxaline-2,3-dione↗