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Biomedical subjects

S Inouye

Publications and source records attributed to S Inouye.

At least 19 recordsLinked to original sources

Cloning and characterization of the homologous genes of firefly luciferase in the mealworm beetle, Tenebrio molitor.

Three homologous genes of firefly luciferase were cloned from the non-luminous beetle Tenebrio molitor. Three gene products for homologues, TmLL-1, TmLL-2 and TmLL-3, showed fatty acyl-coenzyme A (acyl-CoA) synthetic activity, but not luciferase activity with firefly luciferin. The transcripts were detected through the developmental stages in T. molitor. These results suggested that firefly luciferase was evolved from a fatty acyl-coenzyme A synthetase by gene duplications in the insect.

Amino Acid Sequence↗

Direct nondestructive imaging of magnetization in a spin-1 Bose-Einstein gas.

Polarization-dependent phase-contrast imaging is used to resolve the spatial magnetization profile of an optically trapped ultracold gas. This probe is applied to Larmor precession of degenerate and nondegenerate spin-1 87Rb gases. Transverse magnetization of the Bose-Einstein condensate persists for the condensate lifetime, with a spatial response to magnetic field inhomogeneities consistent with a mean-field model of interactions. In comparison, the magnetization of the non-condensed gas decoheres rapidly. Rotational symmetry implies that the Larmor frequency of a spinor condensate be density independent, and thus suitable for precise magnetometry with high spatial resolution.

Journal Article↗

Retrons, msDNA, and the bacterial genome.

Retrons are distinct DNA sequences that code for a reverse transcriptase (RT) similar to the RTs produced by retroviruses and other types of retroelements. Retron DNAs are commonly associated with prophage DNA and are found in the genomes of a wide variety of different bacteria. The retron RT is used to synthesize a strange satellite DNA known as msDNA. msDNA is actually a complex of DNA, RNA, and probably protein. It is composed of a small, single-stranded DNA, linked to a small, single-stranded RNA molecule. The 5' end of the DNA molecule is joined to an internal guanosine residue of the RNA molecule by a unique 2'-5' phosphodiester bond. msDNA is produced in many hundreds of copies per cell, but its function remains unknown. Although retrons are absent from the genome of most members of a population of related bacteria, retrons may not be entirely benign DNAs. Evidence is beginning to suggest that retron elements may produce small but potentially significant effects on the host cell. This includes the generation of repeated copies of the msDNA sequence in the genome, and increasing the frequency of spontaneous mutations. Because these events involve the retron RT, this may represent a source of reverse transcription in the bacterial cell. Thus, the process of reverse transcription, a force that has profoundly affected the content and structure of most eukaryotic genomes, may likewise be responsible for changes in some prokaryotic genomes.

DNA Transposable Elements↗

Observation of heteronuclear Feshbach resonances in a mixture of bosons and fermions.

Three magnetic-field induced heteronuclear Feshbach resonances were identified in collisions between bosonic 87Rb and fermionic 40K atoms in their absolute ground states. Strong inelastic loss from an optically trapped mixture was observed at the resonance positions of 492, 512, and 543+/-2 G. The magnetic-field locations of these resonances place a tight constraint on the triplet and singlet cross-species scattering lengths, yielding (-281+/-15)a(0) and (-54+/-12)a(0), respectively. The width of the loss feature at 543 G is 3.7+/-1.5 G wide; this broad Feshbach resonance should enable experimental control of the interspecies interactions.

Journal Article↗

Sodium Bose-Einstein condensates in the F = 2 state in a large-volume optical trap.

We have investigated the properties of Bose-Einstein condensates of sodium atoms in the upper hyperfine ground state. Condensates in the high-field seeking [F=2, m(F)=-2> state were created in a large volume optical trap from initially prepared [F=1, m(F)=-1> condensates using a microwave transition at 1.77 GHz. We found condensates in the stretched state [F=2, m(F)=-2> to be stable for several seconds at densities in the range of 10(14) atoms/cm(3). In addition, we studied the clock transition [F=1, m(F)=0> --> [F=2, m(F)=0> in a sodium Bose-Einstein condensate and determined a density-dependent frequency shift of (2.44+/-0.25+/-0.5) x 10(-12) Hz cm(3).

Journal Article↗

Analysis of sequential immunoglobulin E-binding epitope of Japanese cedar pollen allergen (Cry j 2) in humans, monkeys and mice.

BACKGROUND: Japanese cedar (Cryptomeria japonica; CJ) pollinosis has been reported to occur naturally in Japanese monkeys (Macaca fuscata) as well as in humans. Most human patients and monkeys with pollinosis have specific IgE for Cry j 2, a major allergen of CJ pollen. OBJECTIVE: The main purpose of this study was to identify IgE B cell epitopes of Cry j 2 using a synthetic peptide in humans, monkeys and mice. METHODS: We synthesized 38 overlapping peptides that span the entire length of Cry j 2. We examined the B cell epitopes of Cry j 2 that are recognized by IgE in the sera of human patients and monkeys with pollinosis and immunized mice using synthetic peptides of Cry j 2. We also examined the reaction of Cry j 2-specific mouse monoclonal IgG antibodies to the peptides. Furthermore, we conducted a histamine release assay with leucocytes from a pollinosis patient using human serum albumin (HSA) conjugated with the peptides as a B cell epitope. RESULTS: We found that 16 of the 20 pollinosis patients who had specific IgE to Cry j 2 also exhibited IgE reaction with some Cry j 2 peptides. Of these 16 patients, 10 exhibited IgE reaction with Cry j 2 peptide no. 13 (121GQCKWVNGREICNDRDRPTA140). Five of the seven monkeys with CJ pollinosis exhibited a reaction with peptide no. 13. Furthermore, IgE in mice immunized with Cry j 2 and two mouse monoclonal IgG antibodies reacted with peptide no. 13. Peptide no. 13-conjugated HSA showed the release of histamine from basophils. Furthermore, to determine the minimum epitope in peptide no. 13, we conducted an enzyme-linked immunosorbent assay inhibition test. The core of the epitope in humans, monkeys and mice was 124KWVNGREI131. CONCLUSION: We found that 124KWVNGREI131 is an important B cell epitope recognized by IgE in humans, monkeys and mice.

Animals↗

Analysis of HLA in children with gelatin allergy.

Systemic immediate reactions including anaphylaxis to gelatin in vaccines have been reported. However, the number of such reports is very small compared with the number of children exposed to gelatin. The present study was designed to investigate whether susceptibility or resistance to gelatin allergy is associated with human leukocyte antigen (HLA) class II gene. Blood samples were obtained from 49 patients with gelatin allergy and specific IgE to gelatin. DNA-based HLA class II typing was performed to determine the DRB1, DQB1 and DPB1 alleles. Genotype frequencies were compared with those found in 240 unrelated controls. The frequency of DQB1*0303 (55.1%) was significantly higher in the patients than in the control subjects (32.1%). The frequency of DPB1*0402 was also significantly higher in the patients (32.7%) than in the control subjects (15.4%). On the other hand, the frequency of subjects carrying DRB1*15 (DRB1*1501 and DRB1*1502) was significantly lower among the patients group (18.4%) than among the controls (40.8%). We found that DQB1*0303 and DPB1*0402 were positively associated with the IgE response for gelatin, while DRB1*15 was negatively associated with it.

Child↗

Realization of Bose-Einstein condensates in lower dimensions.

Bose-Einstein condensates of sodium atoms have been prepared in optical and magnetic traps in which the energy-level spacing in one or two dimensions exceeds the interaction energy between atoms, realizing condensates of lower dimensionality. The crossover into two-dimensional and one-dimensional condensates was observed by a change in aspect ratio and by the release energy converging to a nonzero value when the number of trapped atoms was reduced.

Journal Article↗

Observation of vortex phase singularities in Bose-Einstein condensates.

We have observed phase singularities due to vortex excitation in Bose-Einstein condensates. Vortices were created by moving a laser beam through a condensate. They were observed as dislocations in the interference fringes formed by the stirred condensate and a second unperturbed condensate. The velocity dependence for vortex excitation and the time scale for re-establishing a uniform phase across the condensate were determined.

Journal Article↗

The role of nitric oxide in the gastric acid secretion induced by ischemia-reperfusion in the pylorus-ligated rat.

In a rat model of the ischemia-reperfusion with pylorus ligation, gastric ulcer was formed, although gastric acid secretion was reduced. When the polymorphonuclear leukocytes were inactivated in advance, gastric ulcer was not formed, but acid secretion was increased, indicating that gastric acid is not a cause of the ulcer formation in this model. The mechanism of gastric acid suppression accompanied by ischemia-reperfusion was examined in relation to the role of oxygen-free radicals in this rat model. Prior administration of superoxide dismutase did not modulate acid secretion, but N-nitro-L-arginine methyl ester (L-NAME) increased acid secretion. The action of L-NAME was antagonized specifically by L-arginine, but not by D-arginine. S-nitroso-N-acetylpenicillamine did not inhibit basal acid secretion but antagonized the action of L-NAME. Aminoguanidine increased significantly the gastric acid output that was suppressed by ischemia-reperfusion. When polymorphonuclear leukocytes were inactivated by treatment with their antibody, the gastric acid output recovered to the level in the pylorus-ligated rat without ischemia-reperfusion. These results suggested that nitric oxide (NO) produced by the infiltrated polymorphonuclear leukocytes plays an important role in the suppression of acid secretion induced by ischemia-reperfusion.

Animals↗

Does matter wave amplification work for fermions?

We discuss the relationship between bosonic stimulation, density fluctuations, and matter wave gratings. It is shown that enhanced stimulated scattering, matter wave amplification, and atomic four-wave mixing do not require macroscopic occupation of a single quantum state. These processes are in principle possible for fermionic or nondegenerate samples, if they are prepared in a cooperative state. In practice, there are limitations due to short coherence times.

Journal Article↗

Cloning and characterization of adenylate kinase from Chlamydia pneumoniae.

Chlamydiae proliferate only within the infected host cells and are thought to be "energy parasites," because they take up ATP from the host cell as an energy source. In the present study, we isolated from Chlamydia pneumoniae the gene encoding adenylate kinase (AK). Using the enzyme produced in Escherichia coli, its properties were characterized. K(m) values for AMP and for ADP of the purified C. pneumoniae AK (AKcpn) were each 330 microm, which is significantly higher than the reported values of other AKs, whereas K(m) for ATP was 24 microm, which was rather lower than others. AKcpn contains 1 g atom of zinc/mol of 24,000-dalton protein. Mass spectrometric analysis of AKcpn and analysis of properties of mutated AKcpn strongly suggested that zinc is associated with four cysteine residues in the LID domain of the enzyme. The apo-AKcpn that lost zinc retained AK activity, although K(m) for AMP of apo-AKcpn increased about 2-fold and V(max) decreased about one-half from that of holo-AKcpn. The apo-AKcpn was more thermolabile and sensitive to trypsin digestion than the holo-AKcpn. Moreover, the recovery in vitro of the AK activity during the renaturation process of the denatured apo-AKcpn was dependent on zinc. A mutated protein in which cysteine residues in the LID domain were substituted by other amino acids lost both zinc and enzyme activity. The mutated protein was more sensitive to protease than the apo-AKcpn. These results indicate that zinc in AKcpn, although not essential for the catalysis, stabilizes the enzyme and probably plays a crucial role in proper folding of the protein. Furthermore, the catalytic properties of AKcpn suggest a distinctive regulatory mechanism in the metabolism compared with AKs in other organisms.

Adenylate Kinase↗

Analysis of the canine IgE-binding epitope on the major allergen (Cry j 1) of Japanese cedar pollen with anti-Cry j 1 monoclonal antibodies.

In our previous study [Immunology 91 (1997) 161] using monoclonal antibodies (mAbs) specific to Cry j 1, a major allergen in Japanese cedar (Cryptomeria japonica) pollen, we identified five independent epitopes (EP-1-EP-5) on the molecule and found that EP-1 and EP-5 are the predominant allergic epitopes for humans and monkeys, respectively. In this study, we analyzed the epitopes recognized by IgE in the sera of 10 dogs sensitive to C. japonica pollen allergen using an IgE-ELISA inhibition method with these mAbs. The IgE reaction patterns varied among dogs. In eight of the 10 dogs, IgE recognized EP-5 which is a predominant allergic epitope for monkeys with the pollenosis. In four dogs, IgE recognized EP-1 which is a predominant allergic epitope for human patients with the pollenosis. In three dogs, IgE recognized EP-4 which is a heat-stable epitope. EP-5 is a predominant allergic epitope for dogs and some, but not all, dogs have IgE reaction patterns to the epitopes similar to those of humans.

Allergens↗

Screening of the antibacterial effects of a variety of essential oils on respiratory tract pathogens, using a modified dilution assay method.

The purpose of this study was to examine the antibacterial effects of a wide variety of essential oils on major respiratory tract pathogens. The antibacterial activity of 14 essential oils and their major components was evaluated by agar-plate dilution assay under sealed conditions, with agar used as a stabilizer for homogeneous dispersion. Of the selected strains of four major bacteria causing respiratory tract infection, Haemophilus influenzae was most susceptible to the essential oils, followed by Streptococcus pneumoniae and Streptococcus pyogenes. Staphylococcus aureus was less susceptible. No cross-resistance was observed between penicillin-sensitive and penicillin-resistant S. pneumoniae. Escherichia coli, used as a control bacterium, showed the lowest susceptibility. Essential oils containing aldehyde or phenol as a major component showed the highest antibacterial activity, followed by the essential oils containing terpene alcohols. Other essential oils, containing terpene ketone, or ether, had much weaker activity, and an oil containing terpene hydrocarbon was inactive. Based on these findings, thyme (wild, red, and geraniol types), cinnamon bark, lemongrass, perilla, and peppermint oils were selected for further evaluation of their effects on respiratory tract infection.

Agar↗

The msDNAs of bacteria.

msDNAs are small, structurally unique satellite DNAs found in a number of Gram-negative bacteria. Composed of hundreds of copies of single-stranded DNA--hence the name multicopy single-stranded DNA--msDNA is actually a complex of DNA, RNA, and probably protein. These peculiar molecules are synthesized by a reverse transcription mechanism catalyzed by a reverse transcriptase (RT) that is evolutionarily related to the polymerase found in the HIV virus. The genes, including the RT gene, responsible for the synthesis of msDNA are encoded in a retron, a genetic element that is carried on the bacterial chromosome. The retron is, in fact, the first such retroelement to be discovered in prokaryotic cells. This report is a comprehensive review of the many interesting questions raised by this unique DNA and the fascinating answers it has revealed. We have learned a great deal about the structure of msDNA: how it is synthesized, the structure and functions of the RT protein required to make it, its effects on the host cell, the retron element that encodes it, its possible origins and evolution, and even its potential usefulness as a practical genetic tool. Despite the impressive gains in our understanding of the msDNAs, however, the simple, fundamental question of its natural function remains an enduring mystery. Thus, we have much more to learn about the msDNAs of bacteria.

Bacteria↗

Specific IgE and IgG to gelatin in children with systemic cutaneous reactions to Japanese encephalitis vaccines.

BACKGROUND: Systemic allergic reactions to Japanese encephalitis (JE) vaccine that include urticaria, angioedema, and rash have been reported. In Japan, children who suffered from allergic immediate-type reactions to JE vaccine had antigelatin IgE in their sera. However, the immunologic mechanism of allergic nonimmediate-type reactions that consist of cutaneous signs appearing several hours or more after JE vaccination has not been defined. METHODS: Serum samples were taken from 28 children who showed allergic nonimmediate-type cutaneous reactions to JE vaccine. Furthermore, serum samples were taken from 10 children who showed allergic immediate-type reactions with cutaneous signs and/or respiratory symptoms to JE vaccine. We have defined an immediate-type reaction as one occurring within 1 h after vaccination. RESULTS: Of 10 children who showed immediate-type reactions, all had antigelatin IgE and IgG. Of 28 children who showed systemic nonimmediate-type reactions, one had antigelatin IgE and nine (32%) had antigelatin IgG. The child who had antigelatin IgE showed urticaria 2 h after JE vaccination. CONCLUSION: These results suggest that some children who showed allergic nonimmediate-type reactions to JE vaccine were sensitized to gelatin.

Antibody Specificity↗