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Detecting proteins at low concentrations in high-ionic-strength conditions by silicon nanowire field-effect transistors (SiNWFETs) is severely hindered due to the weakened signal, primarily caused by screening effects. In this study,... more
Detecting proteins at low concentrations in high-ionic-strength conditions by silicon nanowire field-effect transistors (SiNWFETs) is severely hindered due to the weakened signal, primarily caused by screening effects. In this study, aptamer as a signal amplifier, which has already been reported by our group, is integrated into SiNWFET immunosensors employing antigen-binding fragments (Fab) as the receptors to improve its detection limit for the first time. The Fab-SiNWFET immunosensors were developed by immobilizing Fab onto Si surfaces modified with either 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde (GA) (Fab/APTES-SiNWFETs), or mixed self-assembled monolayers (mSAMs) of polyethylene glycol (PEG) and GA (Fab/PEG-SiNWFETs), to detect the rabbit IgG at different concentrations in a high-ionic-strength environment (150 mM Bis-Tris Propane) followed by incubation with R18, an aptamer which can specifically target rabbit IgG, for signal enhancement. Empirical results reveal...
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PROBLEM TO BE SOLVED: To obtain the subject new substance in a vaculovirus expression sys tem, consisting of biologically active humanNGF or a derivative therefrom with variation in the amino acrid sequence while retaining the same... more
PROBLEM TO BE SOLVED: To obtain the subject new substance in a vaculovirus expression sys tem, consisting of biologically active humanNGF or a derivative therefrom with variation in the amino acrid sequence while retaining the same activity as that of biologically active humanNGF, and useful for e.g. treating neuropathy. SOLUTION: This new substance is a biologically active humanNGF or a derivative therefrom through addition, insertion, deletion or substitution of amino acid(s) to, into, from or with the humanNGF while retaining essentially the same activity as the humanNGF, being useful for e.g medicinal compositions for treating neuropathy such as Alzheimer's disease. This biologically active humanNGF is obtained by the following process: a human leukocyte genome library is subjected to Southern blotting analysis using a terminal-labeled synthetic oligonucleotide complementary to the fragment of a humanNGF gene as a hybridization probe to select positive clone, its DNA is then...
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ES provided a method for the construction of DNA molecules RECOMBINANT FACTOR CAPABLE OF PRODUCING A NERVE GROWTH OF HUMAN biologically active (hNGF) in insect cells. MATURE protein expression TRIGGERED USandA BACULOVIROSA expression... more
ES provided a method for the construction of DNA molecules RECOMBINANT FACTOR CAPABLE OF PRODUCING A NERVE GROWTH OF HUMAN biologically active (hNGF) in insect cells. MATURE protein expression TRIGGERED USandA BACULOVIROSA expression system. Biologically active hNGF thus produced is a value POTENTIAL IN THE TREATMENT OF PATIENTS SUFFERING FROM DISEASE AND OTHER ALZHEIMAR Neurological Disorders.
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Transcription of the interleukin 1 beta (IL-1 beta) gene was studied by mRNA hybridization with a cDNA probe in the human promonocytic cell line U-937. Phorbol ester and lipopolysaccharide increased the steady-state level of IL-1 beta... more
Transcription of the interleukin 1 beta (IL-1 beta) gene was studied by mRNA hybridization with a cDNA probe in the human promonocytic cell line U-937. Phorbol ester and lipopolysaccharide increased the steady-state level of IL-1 beta mRNA. Glucocorticoids markedly decreased IL-1 beta mRNA levels by two mechanisms. Transcription of the IL-1 gene was inhibited, as shown by in vitro transcription assays with nuclei isolated from glucocorticoid-treated cells. Moreover, kinetic analyses and pulse-labeling of mRNAs showed that glucocorticoids selectively decrease the stability of IL-1 beta mRNA, without affecting the stability of beta-actin and FOS mRNAs. Inhibition of the formation and effects IL-1 is a mechanism by which glucocorticoids can exert antiinflammatory and immunosuppressive effects.
Research Interests: Molecular Biology, Kinetics, Biology, Medicine, Gene expression, and 15 moreMultidisciplinary, DNA, Lipopolysaccharide, Dexamethasone, Cell line, Humans, Nucleic acid hybridization, PNAS, Glucocorticoid, Glucocorticoids, Phorbol ester, Interleukin, In Vitro Transcription, Cycloheximide, and hydrocortisone
A DNA-transfection protocol has been developed that makes use of a synthetic cationic lipid, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA). Small unilamellar liposomes containing DOTMA interact spontaneously with... more
A DNA-transfection protocol has been developed that makes use of a synthetic cationic lipid, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA). Small unilamellar liposomes containing DOTMA interact spontaneously with DNA to form lipid-DNA complexes with 100% entrapment of the DNA, DOTMA facilitates fusion of the complex with the plasma membrane of tissue culture cells, resulting in both uptake and expression of the DNA. The technique is simple, highly reproducible, and effective for both transient and stable expression of transfected DNA. Depending upon the cell line, lipofection is from 5- to greater than 100-fold more effective than either the calcium phosphate or the DEAE-dextran transfection technique.
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Background Human β-amyloid, the main component in the neuritic plaques found in patients with Alzheimer's disease, is generated by cleavage of the β-amyloid precursor protein. Beyond the role in pathology, members of this protein... more
Background Human β-amyloid, the main component in the neuritic plaques found in patients with Alzheimer's disease, is generated by cleavage of the β-amyloid precursor protein. Beyond the role in pathology, members of this protein family are synaptic proteins and have been associated with synaptogenesis, neuronal plasticity and memory, both in vertebrates and in invertebrates. Consolidation is necessary to convert a short-term labile memory to a long-term and stable form. During consolidation, gene expression and de novo protein synthesis are regulated in order to produce key proteins for the maintenance of plastic changes produced during the acquisition of new information. Results Here we partially cloned and sequenced the beta-amyloid precursor protein like gene homologue in the crab Chasmagnathus (cappl), showing a 37% of identity with the fruit fly Drosophila melanogaster homologue and 23% with Homo sapiens but with much higher degree of sequence similarity in certain regions...
Research Interests: Cognitive Science, Biology, Immunohistochemistry, Drosophila melanogaster, Medicine, and 15 moreGene expression, Animals, Male, Homo Sapiens, Central Nervous System, Immunoprecipitation, Brachyura, Associative Memory, Long Term Memory, Fruit Fly, Amino Acid Sequence, Base Sequence, Biochemistry and cell biology, Amyloid Beta Precursor Protein, and BMC
L'invention decrit l'hydrolyse enantioselective d'esters de naproxene racemiques par des esters hydrolases, les hydrolases etant derivees d'un groupe de micro-organismes dont le plus approprie etait le micro-organisme,... more
L'invention decrit l'hydrolyse enantioselective d'esters de naproxene racemiques par des esters hydrolases, les hydrolases etant derivees d'un groupe de micro-organismes dont le plus approprie etait le micro-organisme, Zopfiella latipes. Les esters hydrolases sont utilises dans une hydrolyse d'esters de naproxene racemiques dont le cout est faible et le rendement eleve.
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PURPOSE: To obtain a novel biologically active human nerve growth factor(hNGF) and a derivative thereof for the treatment of human neurological disorders such as Alzheimer's disease or the like, which is obtained by expressing a... more
PURPOSE: To obtain a novel biologically active human nerve growth factor(hNGF) and a derivative thereof for the treatment of human neurological disorders such as Alzheimer's disease or the like, which is obtained by expressing a recombinant DNA of hNGF in insect cells using a baculovirus expression system. CONSTITUTION: The novel biologically active hNGF and a derivative thereof useful as a therapeutic agent for human neurological disorders such as Alzheimer's disease or the like can be obtained by infecting insect cells such as Spodoptera-frugiperda(Sf9) with a baculovirus transfer vector having been integrated with the gene of a human βNGF(hNGF) comprising the prepro-portion of the DNA coding sequence of mouse nerve growth factor(NGF) and the DNA coding sequence of maturation human βNGF(hNGF) or the prepro-portion of the DNA coding sequence of human NGF and the DNA coding sequence of maturation human βNGF(hNGF) or a derivative thereof which has been bonded to the controlli...
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About 14.3, 20.4, and 23.0 ± that the crystalline hydrochloride Raney set theory and a method with the features are provided in a powder x- ray diffraction pattern of 0.2 ° with the peak at 2-theta.
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The biological activity of polyoma viral DNA was evaluated in mice and hamsters. Viral DNA administered parenterally is about 4 to 5 logs less efficient than polyoma virions in establishing infection in mice. Supercoiled viral DNA was... more
The biological activity of polyoma viral DNA was evaluated in mice and hamsters. Viral DNA administered parenterally is about 4 to 5 logs less efficient than polyoma virions in establishing infection in mice. Supercoiled viral DNA was infectious for mice after parenteral administration, giving mean infective doses of 10(-3) to 10(-4) microgram. However, animals fed microgram quantities of polyoma DNA I did not become infected. Linearization of viral DNA with R.EcoRI or R.BamHI, which are single-cut enzymes cleaving in the early and late regions of the genome, respectively, reduced the infectivity for mice approximately fivefold. Approximately 10% of newborn hamsters inoculated intraperitoneally with polyoma DNA I developed tumors. In contrast, the same amount of viral DNA which had been cleaved in the early region with R.EcoRI induced tumors in 50% of inoculated hamsters.
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The internal organization of endogenous xenotropic murine leukemia virus proviruses was determined in a series of blot hybridization experiments in which DNA from several different inbred mouse strains, digested with restriction enzymes... more
The internal organization of endogenous xenotropic murine leukemia virus proviruses was determined in a series of blot hybridization experiments in which DNA from several different inbred mouse strains, digested with restriction enzymes known to cleave xenotropic proviral DNAs at least twice, was annealed to generalized murine leukemia virus or xenotropic env-specific DNA probes. Comigrating bands of variable intensity which hybridized to the xenotropic env probe were identified in all inbred mouse DNA preparations. At least seven classes of endogenous xenotropic proviral DNA with respect to SacI cleavage maps were detected in mouse DNA. Two of the seven classes were indistinguishable from proviruses associated with known infectious xenotropic murine leukemia viruses. These results are consistent with the existence of related but organizationally distinct families of endogenous xenotropic proviral DNA that are present in different relative abundances in mouse genomic DNA.
Research Interests: International organizations, Biology, Virology, Medicine, Biological Sciences, and 11 moreMice, Animals, Nucleic acid hybridization, Enzyme, Genetic Recombination, International Organizations, Relative Abundance, Base Sequence, Virus isolation, DNA Restriction Enzymes, and Medical and Health Sciences
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A series of recombinant plasmids containing polyoma virus (PY) DNA were constructed, and their biological activity was evaluated in mice and in cultured mouse cells. While all of the recombinants studied contain the complete, potentially... more
A series of recombinant plasmids containing polyoma virus (PY) DNA were constructed, and their biological activity was evaluated in mice and in cultured mouse cells. While all of the recombinants studied contain the complete, potentially infectious viral DNA, in no case was the intact recombinant PY-plasmid DNA, or live Escherichia coli containing the recombinant plasmids, capable of inducing PY infection of mice, either by feeding or by parenteral injection.
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Increasing recognition of diffuse plaques has raised questions about the differences between diffuse and neuritic plaques, particularly in regard to the role of amyloid precursor protein (APP) processing in their formation. To address... more
Increasing recognition of diffuse plaques has raised questions about the differences between diffuse and neuritic plaques, particularly in regard to the role of amyloid precursor protein (APP) processing in their formation. To address this issue, corpus striatum (containing almost exclusively diffuse plaques) and cerebral cortex (containing an admixture of plaque types) from patients with Alzheimer's disease (AD) were examined immunohistochemically with antibodies to domain-specific sites of APP (N-terminal, C-terminal, beta A4-related, isoform-specific, and other epitopes). Striatal plaques labeled strongly with beta A4 antibodies as did cortical plaques in AD and the occasional diffuse plaques in cortex from nondemented elderly controls. Weak labeling of some cortical neuritic plaques but not diffuse plaques was observed with antibodies directed against other APP epitopes. Electron microscopy of diffuse plaque-rich striatum in AD cases revealed only rare degenerating neurites without apparent fibrillar amyloid; no changes were noted in the plaque-free striatum of controls. These results suggest that antibodies to beta A4 recognize not only fibrillar amyloid of neuritic plaques but also antigenic determinants of diffuse plaques which lack fibrillar amyloid. Furthermore, the finding that antibodies to non-A4 domains of APP labeled only cortical but not striatal plaques suggests that APP processing mechanisms in cortical and striatal tissues may differ.
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A cDNA clone (designated as GP2-7) encoding a novel 5-hydroxytryptamine (5-HT) receptor was isolated from a guinea pig hippocampal library. The receptor shares amino acid homology within the hydrophobic domains with other cloned 5-HT... more
A cDNA clone (designated as GP2-7) encoding a novel 5-hydroxytryptamine (5-HT) receptor was isolated from a guinea pig hippocampal library. The receptor shares amino acid homology within the hydrophobic domains with other cloned 5-HT receptor subtypes (34-48%). The sequence of GP2-7 is homologous to that described for a novel receptor previously cloned from a rat brain cDNA library and provisionally designated as 5-HT7. mRNA for GP2-7 was detected in cortical and limbic brain regions. Transiently expressed GP2-7 showed high-affinity binding to [3H]5-HT (pKi = 9.0) with the following rank order of affinities: 5-carboxyamidotryptamine (5-CT) > 5-HT = 5-methoxytryptamine (5-MeOT) > methiothepin > 8-hydroxy-2-(dipropylamino)tetralin (8-OH-DPAT) > spiperone > sumatriptan. Adenylyl cyclase activity in CHO-K1 cells transiently transfected with GP2-7 was stimulated by several analogues of 5-HT with the following order of potency: 5-CT > 5-HT = 5-MeOT > dipropyl-5-CT > 8-OH-DPAT. Methiothepin and spiperone were potent antagonists. Preliminary analysis suggests that GP2-7 closely resembles a receptor in the guinea pig hippocampus that exhibits a high affinity toward 5-CT.
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Page 1. J Mol Neurosci (1990) 2:19-27 Journal of Molecular Neuroscience 9 Birkh~iuser Boston 1990 Processing of Alzheimer's Disease-Associated [3-Amyloid Precursor Protein Nazneen N. Dewji, 1 Earl R. Shelton, 2 Mark ...
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Research Interests: Biology, Biological Chemistry, Medicine, Biological Sciences, Neuropeptides, and 15 moreHumans, Animals, Biological, Polymerase Chain Reaction, Neurons, CHEMICAL SCIENCES, Dorsal root ganglia, Molecular cloning, Rats, Tetrodotoxin, Amino Acid Sequence, Oocytes, Molecular Sequence Data, sodium channels, and Medical and Health Sciences
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Research Interests: Biology, Medicine, Quantitative PCR, Humans, Animals, and 15 moreMale, Polymerase Chain Reaction, Ligand Binding, Prostate, Molecular cloning, Human Genome, Radioligand Assay, Amino Acid Sequence, Base Sequence, Biochemistry and cell biology, Molecular Sequence Data, Inositol Phosphates, CHO cells, Cricetinae, and Chinese hamster ovary
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Escherichia coli expression vectors encoding an acid-labile aspartyl-proline (Asp-Pro) dipeptide bridging two protein sequences were constructed and used to synthesize two different bovine growth hormone (bGH) fusion proteins. The codons... more
Escherichia coli expression vectors encoding an acid-labile aspartyl-proline (Asp-Pro) dipeptide bridging two protein sequences were constructed and used to synthesize two different bovine growth hormone (bGH) fusion proteins. The codons GAT-CCX coding for Asp-Pro are provided by the recognition site for Bam HI (GGATCC). Treatment of the bGH fusion proteins at low pH in the presence of guanidine hydrochloride releases the bGH moiety from the fusion protein. The release of the bGH from the fusion protein specifically requires the Asp-Pro dipeptide linking the bGH sequence to the fusion protein. The bGH released from the fusion protein retains anti-bGH immunoreactivity as well as the ability to bind to growth hormone receptor in vitro.
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The biological activity of polyoma viral DNA was evaluated in mice and hamsters. Viral DNA administered parenterally is about 4 to 5 logs less efficient than polyoma virions in establishing infection in mice. Supercoiled viral DNA was... more
The biological activity of polyoma viral DNA was evaluated in mice and hamsters. Viral DNA administered parenterally is about 4 to 5 logs less efficient than polyoma virions in establishing infection in mice. Supercoiled viral DNA was infectious for ...