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Biomedical subjects

Jun Kobayashi

Publications and source records attributed to Jun Kobayashi.

At least 37 records · Page 2Linked to original sources

Phantom encodes the 25-hydroxylase of Drosophila melanogaster and Bombyx mori: a P450 enzyme critical in ecdysone biosynthesis.

We have reported recently the identification and characterization of the last three mitochondrial cytochrome P450 enzymes (CYP) controlling the biosynthesis of 20-hydroxyecdysone, the molting hormone of insects. These are encoded by the following genes: disembodied (dib, Cyp302a1, the 22-hydroxylase); shadow (sad, Cyp315a1, the 2-hydroxylase); and shade (shd, Cyp314a1, the 20-hydroxylase). Employing similar gene identification and transfection techniques and subsequent biochemical analysis of the expressed enzymatic activity, we report the identity of the Drosophila gene phantom (phm), located at 17D1 of the X chromosome, as encoding the microsomal 25-hydroxylase (Cyp306a1). Similar analysis following differential display-based gene identification has also resulted in the characterization of the corresponding 25-hydroxylase gene in Bombyx mori. Confirmation of 2,22,25-trideoxyecdysone (3beta,5beta-ketodiol) conversion to 2,22-dideoxyecdysone (3beta,5beta-ketotriol) mediated by either Phm enzyme employed LC, MS and definitive NMR analysis. In situ developmental gene analysis, in addition to northern, western and RT-PCR techniques during Drosophila embryonic, larval and adult development, are consistent with this identification. That is, strong expression of phm is restricted to the prothoracic gland cells of the Drosophila larval ring gland, where it undergoes dramatic changes in expression, and in the adult ovary, but also in the embryonic epidermis. During the last larval-larval transition in Bombyx, a similar expression pattern in the prothoracic gland is observed, but as in Drosophila, slight expression is also present in other tissues, suggesting a possible additional role for the phantom enzyme.

Amino Acid Sequence↗

Force deficits in skeletal muscle after delayed reinnervation.

Using a rat hindlimb model, the authors tested the hypothesis that, in muscles reinnervated after long-term denervation, atrophy-dependent and atrophy-independent mechanisms operate independently to produce force deficits. In adult rats, gastrocnemius muscles were subjected to denervation via tibial nerve transection. Reconstruction of the nerve lesion was delayed for periods ranging from 2 weeks to 1 year. After a minimum recovery period of 6 months after nerve repair, muscle mass and maximum isometric tetanic force were measured and specific force was calculated for each muscle (n = 40 muscles from 23 animals). After recovery, observed deficits in muscle mass and maximum tetanic force were directly proportional to the denervation interval. On the other hand, the deficit in specific force was not proportional to the denervation interval; all groups in which the nerve reconstruction was delayed for a month or longer demonstrated a deficit of 30 percent to 50 percent. These data support our hypothesis that, after prolonged denervation followed by reinnervation, the magnitude of the deficit in whole muscle force does not parallel the deficit in specific force. These data support the idea that mechanisms governing muscle atrophy are independent of those resulting in specific force deficits.

Animals↗

Metal elution and content of fused slags produced from incinerated ash.

In recent years, trials to reuse fused slags, obtained through the further treatment of incinerated ash produced in the process of the disposal of garbage or sewer sludge, such as for building materials including bricks, have drawn attention. To establish the safety of using such substances, elution tests for metals and metal contents were investigated in this study. As the result of examining 13 slags, which were made by different methods, little elution of toxic metals was found in the elution tests when water was used as a solvent. However when acid was used, there was a tendency for the elution of many kinds of metals to become greater. On other hand, the results of content measurements showed that arsenic and selenium, which were not seen to be eluted from any of the slags tested, were detected; and that aluminum and manganese existed in the slags in high concentrations.

Air Pollutants↗

[Physical properties and metal elutions of the thirteen molten slags produced from incinerated ash].

In recent years, there have been many attempts to reuse molten slag. Molten slag is obtained through further treatment of incinerated ash produced in the disposal process of garbage or sewage sludge and is used in building materials, such as brick. To establish the safety of such slag, its physical properties and metal contents were investigated in this study. After examining the physical and chemical characteristics of 13 forms of slag made using different methods, no differences were observed and it was judged that reuse is possible. There was little elution of toxic metals in the elution test when water was used as a solvent. On the other hand, when acid and alkali were used, the elution of toxic metals tended to become greater.

Chemical Phenomena↗

Cross-linked thermoresponsive anionic polymer-grafted surfaces to separate bioactive basic peptides.

Cross-linked, thermoresponsive poly(N-isopropylacrylamide-co-acrylic acid-co-N-tert-butylacrylamide) [poly(IPAAm-co-AAc-co-tBAAm)] thin hydrogel layers on silica beads were used as new column matrix modifiers for LC separation of basic bioactive peptides, angiotensin subtypes I, II, and III. Terpolymer poly(IPAAm-co-AAc-co-tBAAm) showed both phase transition and apparent carboxylate pKa shifts in water, depending on temperature. Polymer-grafted silica bead surfaces exhibited simultaneous thermally modulated changes in hydrophilic/hydrophobic properties and charge densities. More effective separation of angiotensin peptide subtypes was achieved on columns of these terpolymer thin hydrogel grafted surfaces, as compared to an uncharged control binary copolymer of IPAAm and tBAAm. Although hydrophobic interactions effect separation of angiotensin subtypes, combined electrostatic and hydrophobic interaction resulted in more pronounced retention. At temperature below the terpolymer phase transition, hydrophobic interactions predominated, and minimal changes in electrostatic interactions were supported by little shift in the apparent AAc carboxylate pKa values. Above the phase transition temperature, electrostatic interactions were dramatically reduced as a result of the decreased charge densities of the polymer grafted surfaces. Therefore, peptide retention times were also reduced, exhibiting a maximum at near 30-35 degrees C. Interestingly, column retention behavior of angiotensins is dramatically modulated by applied step temperature gradients. Thermoresponsive surface property alteration is a very rapid, reversible phenomenon, allowing step temperature gradients on thermoresponsive columns to enable the analogous performance advantages as gradient elution in reversed-phase HPLC. More importantly, injected peptides were recovered completely from the columns from calculation of peak area. In conclusion, these anionic thermoresponsive polymer-modified surfaces are good candidates for improved separation of bioactive peptides under exclusively aqueous conditions.

Amino Acid Sequence↗

New bulky chiral Lewis acid catalyst: 3,3'-di(2-mesitylethynyl)binaphthol-titanium(IV) complex.

A new chiral Lewis acid catalyst 9 was prepared in situ from a 1:2 molar mixture of (R)-3,3'-di(2-mesitylethynyl)binaphthol (6) and titanium(IV) isopropoxide at ambient temperature. The 3- and 3'-substituents on 6 were effective for preventing undesired aggregation between Ti(IV) complexes and increasing the enantioselectivity (up to 82% ee) in the Diels-Alder reaction of methacrolein with cyclopentadiene.

Journal Article↗

Expansion of unconventional T cells with natural killer markers in malaria patients.

Immunological states during human malarial infection were examined. In parallel with parasitemia and anemia, granulocytosis was induced in the blood of patients, especially those infected with Plasmodium (P.) falciparum. At that time, the level of lymphocytes remained unchanged or slightly increased in the blood. However, the distribution of lymphocyte subsets was modulated, showing that the proportion of CD56(+)T cells, CD57(+)T cells, and gammadeltaT cells (i.e. all unconventional T cells) had increased in patients infected with P. falciparum or P. vivax. This phenomenon occurred at the early phase of infection and disappeared in the course of recovery. The data from patients with multiple attacks of P. vivax infection showed that there was no augmentation of these responses. In adult cases, the increase in the proportion of unconventional T cells seemed to closely parallel disease severity. However, all these responses were weak in children, even those infected with P. falciparum. In conjunction with accumulating evidence from mouse malaria experiments, the present results suggest that the immunological state induced by malarial infection might mainly be an event of unconventional T cells and that the immunological memory might not be long-lasting, possibly due to the properties of unconventional T cells.

Adolescent↗

[Successful treatment with cyclosporine in a case of progressive interstitial pneumonia associated with systemic sclerosis].

A 55-year-old man was admitted to our hospital with dyspnea on effort and sclerodactyly. The chest radiograph showed reticulonodular shadows in both lower lung fields. A diagnosis of systemic sclerosis was made based on the characteristic skin lesions, interstitial pneumonia and the positivity for anti-Scl-70 antibody. Surgical biopsies were performed, which showed various features characteristic of organizing pneumonia and non-specific interstitial pneumonia. Prednisolone and cyclophosphamide were given to the patient, but clinical symptoms and chest radiographic findings worsened. After cyclophosphamide was replaced with cyclosporine, his symptoms, chest radiographic findings and pulmonary function improved and stabilized. It is suggested that cyclosporine is useful for the treatment of progressive interstitial pneumonia associated with systemic sclerosis.

Cryptogenic Organizing Pneumonia↗

Field application and evaluation of a rapid immunochromatographic test for detection of Plasmodium falciparum infection among the inhabitants of Lao PDR.

Field application and evaluation of a rapid immunochromatographic test (ICT) for detection of Plasmodium falciparum infection were performed in 13 villages in a southern province of Lao PDR in 1999. More than 2,000 inhabitants, accounting for 61.8% of the total estimated population, were examined. Malaria infection was confirmed in all villages surveyed by ICT and microscopic diagnosis. The positive rates of P. falciparum malaria by microscopy ranged from 9.7% to 59.2% (mean 27.2%), whereas by ICT they were from 11.6% to 64.5% (mean 29.8%). The positive rates by ICT were generally higher in 8 out of 13 villages. However, a significant difference between the positive rates by microscopy and ICT was not observed in all villages. Plasmodium falciparum infection was actually confirmed by microscopy in 84.1% of specimens that tested positive by ICT. The results by ICT were consistent with those of the microscopic diagnosis, the discrepancy of the results was less than 10% (141/2,066). The ICT was falsely-positive in 4.7% and falsely-negative in 2.1% of the test cases. These results showed the efficacy of ICT not only in the diagnosis of the respective cases, but also in the mass-examination in the field.

Animals↗

Asymmetric intramolecular [3 + 2] cycloaddition reactions of acylhydrazones/olefins using a chiral zirconium catalyst.

Catalytic asymmetric intramolecular [3 + 2] cycloaddition of hydrazone/olefins has been attained. In the presence of a chiral zirconium catalyst prepared from zirconium alkoxide and a BINOL derivative, the desired pyrazolidine derivatives were obtained in high yields with high selectivities. The products were easily converted to 1,3-diamine or beta-aminonitrile derivatives by N-N bond cleavage.

Journal Article↗

Catalytic enantioselective addition of propionate units to imines: an efficient synthesis of anti-alpha-methyl-beta-amino acid derivatives.

Optically active anti-alpha-methyl-beta-amino acid derivatives have been prepared based on catalytic enantioselective addition of propionate units to simple and inert imines using a chiral zirconium complex. High reactivity and selectivity with wide substrate scope were attained by using a new chiral ligand, (R)-6,6'-bis(pentafluoroethyl)-1,1'-bi-2-naphthol ((R)-6,6'-C(2)F(5)BINOL). The reactions using geometrically isomeric ketene silyl acetals gave excellent anti-selectivity with high enantiomeric excess in both cases. Synthetic utility of this reaction has been demonstrated by the preparation of various anti-alpha-methyl-beta-amino acid and trans-3,4-disubstituted beta-lactam derivatives.

Amino Acids↗

Ventricular afterload and ventricular work in fontan circulation: comparison with normal two-ventricle circulation and single-ventricle circulation with blalock-taussig shunts.

BACKGROUND: Recent studies have indicated that there are inherent limitations associated with Fontan physiology. However, there have been no quantitative analyses of the effects of right heart bypass on ventricular afterload, hydraulic power, and resultant overall hemodynamics. Methods and Results- During routine cardiac catheterization, aortic impedance and ventricular hydraulic power were determined, both at rest and under increased ventricular work induced by dobutamine, in 17 patients with Fontan circulation, 15 patients with a single ventricle whose pulmonary circulation was maintained only by Blalock-Taussig shunts, and 13 patients who had normal 2-ventricle circulation. Both vascular resistance (nonpulsatile load on the ventricle) and pulsatile components of ventricular afterload (represented by low-frequency impedance) were significantly higher in the Fontan group than in the other groups (P<0.01), and this was associated with decreased cardiac output in the Fontan patients. In addition, hydraulic power cost per unit forward flow was 40% lower in the 2-ventricle circulation than in the single-ventricle circulation, suggesting lower ventricular efficiency in single-ventricle circulation attributable to the lack of a pulmonary ventricle. Furthermore, in the Fontan group, beta-adrenergic reserve was markedly decreased because of a limited preload reserve. CONCLUSIONS: Fontan physiology is associated with disadvantageous ventricular power and afterload profiles and has limited ventricular reserve capacity. Thus, to improve the long-term prognosis of patients after Fontan surgery, future research should be conducted into medical interventions that can overcome these limitations inherent in Fontan circulation.

Adrenergic beta-Agonists↗

Aqueous chromatography utilizing hydrophobicity-modified anionic temperature-responsive hydrogel for stationary phases.

A new pH-/temperature-responsive poly(N-isopropylacrylamide-co-acrylic acid-co-N-tert-butylacrylamide) (poly(IPAAm-co-AAc-co-tBAAm)) hydrogel grafted on silica beads was evaluated as column matrix for a cation-exchange thermoresponsive chromatography. The stationary phase showed simultaneous changes in temperature-responsive surface charge density and hydrophobicity by incorporation of anionic AAc and hydrophobic tBAAm into IPAAm sequences. Thermoresponsive polymer property alterations were confirmed by temperature-responsive phase transition and shift in apparent pKa values. Catecholamine derivatives were retained on poly(IPAAm-co-AAc-co-tBAAm)-modified column at pH 7.0. Analyte retention was primarily due to the electrostatic interaction. It was noted that the temperature-induced phase transition of poly(IPAAm-co-AAc-co-tBAAm) hydrogel layer on the stationary phases was evidenced by the apparent inflection point in van't Hoff plots around 36 degrees C. This suggests that solute interactions should be changed below and above the stationary phase transition temperature, reducing electrostatic interaction above the transition temperature.

Anions↗

EGF and beta1 integrin convergently regulate migration of A431 carcinoma cell through MAP kinase activation.

We found that the convergently epidermal growth factor (EGF)-induced signal and the collagen-induced signal activate mitogen-activated protein kinase (MAPK), which induces migration. We examined the signaling mechanisms of EGF-induced cell migration on collagen using the A431 carcinoma cell. EGF (10 ng/ml) induced migration on collagen, but inhibited proliferation. Using a MAPK cascade inhibitor, PD98059, it was shown that EGF-induced migration on collagen was mediated by MAPK whereas EGF-induced migration on fibronectin and vitronectin was not. PD98059 also showed that activation of MAPK induced by EGF enhanced the adhesiveness of A431 cells to collagen. By Western blotting analysis, the kinetics of MAPK phosphorylation induced by EGF and collagen was examined separately, and convergently. First of all, EGF without collagen caused transient MAPK phosphorylation. Collagen without EGF caused MAPK to be immediately and transiently dephosphorylated, and rephosphorylated followed by sustained hyperphosphorylation. EGF together with collagen caused an immediate, and sustained, hyperphosphorylation. These facts suggest that the transient MAPK dephosphorylation induced by collagen is required for migration in order to maintain an appropriate level of sustained phosphorylation. Furthermore, we found that adhesion of A431 cells to collagen was blocked by the anti-beta1 integrin antibody or by the mixed antibodies composed of anti-alpha1, -alpha2, and -alpha3 antibodies, indicating that collagen-induced MAPK phosphorylation was mediated through alpha1beta1, alpha2beta1, and alpha3beta1 integrins.

Cell Movement↗

Genome mapping and gene analysis of Antheraea pernyi nucleopolyhedrovirus for improvement of baculovirus expression vector system.

We have constructed a genome DNA map of the Antheraea pernyi nucleopolyhedrovirus (AnpeNPV) and used it to identify target genes for deletion in order to improve the newly developed baculovirus expression vector system. Initially, 50 independent PstI fragments of viral DNA were obtained by shotgun cloning, and both termini of each cloned fragment were sequenced. Then, the sequence data were used for homology search against both nucleotide and amino acid sequences of other NPVs in databases. This homology search allowed us to construct a nearly complete restriction map of a viral DNA with several assumed gaps. Four additional PstI fragments covering the gaps were obtained by PCR amplification, and a complete map of a circular viral DNA, which consisted of 54 PstI fragments, was constructed. The map indicated that the AnpeNPV genome is approximately 130.2 kbp in size and possesses high similarity to the Orgyia pseudotsugata multicapsid NPV (OpMNPV) genome in both sequence and arrangement of genes. Utilizing the genome-wide high similarity between AnpeNPV and OpMNPV, we identified two target genes on the map, namely, cathepsin and chitinase genes, whose products have been proved to be involved in the degradation of recombinant proteins and the liquefaction of virus-infected insect tissues. Comparative sequence analysis of the map also revealed the lack of certain OpMNPV open reading frame (ORF) homologs and the presence of ORFs, whose homologs do not exist in OpMNPV but in other group I NPVs, providing an insight into the position of AnpeNPV in the baculovirus phylogeny.

Journal Article↗

An ecological survey of dengue vector mosquitos in central Lao PDR.

An ecological survey of dengue vector mosquitos was carried out in June 2000 in central Lao PDR. Two areas in Khammouane Province, Nongbok and Thakhek, were selected for the survey. Of the 7 mosquito species identified, Aedes aegypti was dominant in both study areas. The container index for Ae. aegypti in Nongbok was 51.8% and was significantly higher than that of Thakhek (40.2%); moreover, significant differences between the study areas were found with records to containers and to the conditions surrounding the houses. The key containers in Nongbok were water jars, whereas drums or small or discarded containers had the highest occurrence rate of Ae. aegypti in Thakhek. Mesocyclops aspericornis was found in large water jars and cement water tanks; no Aedes larvae were found at these sites. Strategy to control dengue vectors in the study areas was discussed.

Aedes↗

Knowledge and behavior relating to malaria in malaria endemic villages of Khammouane Province, Lao PDR.

In order to provide basic data for evaluation of malaria control measures, a study on the knowledge and behavior of people regarding prevention of malaria was carried out in 8 malaria endemic villages in Khammouane Province of Lao PDR from 1999 to 2000. The total valid questionnaire respondents were 932, with a mean age of 32.3 +/- 14.9. 43.7% of the respondents were illiterate. About 44% of the respondents suffered from malaria in the past. About 55% of the illiterate group slept in mosquito nets, compared to 75.4% for the educated group. About 29% of the illiterate respondents had knowledge of malaria transmission by mosquito bites, compared to 48.8% for the educated groups. Out of 167 non-impregnated mosquito nets examined in two villages, 13 were in bad condition having holes or leaks and 39 female mosquitos including Anopheles spp were collected in these nets by early morning catches. Knowledge of malaria and behavior in relation to the prevention of malaria were significantly related to educational level. Health education as well as general education must be taken into account for communities in malaria endemic areas to become more involved in malaria control strategies.

Adult↗