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Biomedical subjects

T Nagai

Publications and source records attributed to T Nagai.

At least 163 records · Page 9Linked to original sources
PubMed2000

Both fertilization promoting peptide and adenosine stimulate capacitation but inhibit spontaneous acrosome loss in ejaculated boar spermatozoa in vitro.

Both fertilization promoting peptide (FPP) and adenosine stimulate capacitation and inhibit spontaneous acrosome loss in epididymal mouse spermatozoa; these responses involve modulation of the adenylyl cyclase (AC)/cAMP signal transduction pathway. However, it was unclear whether these responses were restricted to the mouse or possibly common to many mammalian species. To address this question, the response of boar spermatozoa to FPP and/or adenosine was evaluated. FPP is found in nanomolar concentrations in seminal plasma of several mammals, but not the pig. When cultured in caffeine-containing Medium 199 for 2 hr, chlortetracycline fluorescence evaluation indicated that neither FPP nor adenosine stimulated boar sperm capacitation per se but did inhibit spontaneous acrosome loss. However, in caffeine-free medium, FPP and adenosine both stimulated capacitation and inhibited spontaneous acrosome loss, suggesting that boar spermatozoa have receptors for both FPP and adenosine. Gln-FPP, a competitive inhibitor of FPP in mouse spermatozoa, has recently been shown to inhibit mouse sperm responses to adenosine as well, suggesting that FPP receptors and adenosine receptors interact in some way. Used with boar spermatozoa, Gln-FPP also significantly inhibited responses to both FPP and adenosine. These responses suggest that mechanisms whereby FPP and adenosine can regulate sperm function, via AC/cAMP, are of considerable physiological significance. Mouse, human, and now boar spermatozoa have been shown to respond to FPP, suggesting that these mechanisms may be common to many mammalian species. We also suggest that the effects of FPP and adenosine could also be exploited to maximize monospermic fertilization in porcine in vitro fertilization.

Acrosome↗
PubMed2000

Evaluation of skin damage caused by percutaneous absorption enhancers using fractal analysis.

Fractal analysis of the cross-sectional morphology of rat skin was conducted to evaluate pathologic changes evoked by percutaneous absorption enhancers. Male hairless rats (WBN/Ht-ILA), 8 weeks old, weighing 160 to 180 g were used. Under anesthetization, glass cells (10-mm inner diameter) were attached to the rats' abdomens, and test solutions containing various mixtures of the percutaneous absorption enhancers, sodium lauryl sulfate, isopropanol, 2-methyl-1-butanol, and sodium myristate were applied. Six hours after application, the solutions were removed and the abdominal skin was excised. Skin cross sections were analyzed with a charge-coupled device (CCD) camera. Image data taken by the CCD camera were fed into a desktop digital computer; then the fractal dimension of each skin cross section was determined on the basis of the box-counting algorithm. A pathologic study was also performed on the skin treated with the test solution. All sections of skin were examined with an optical photo microscope. Pathologic findings were classified into five levels. The total irritation score (TIS) was defined as the summation of damage levels in all regions. Only with the administration of hydrogel containing 2-methyl-1-butanol or sodium lauryl sulfate were positive values of TIS observed. However, the TIS values were independent of the concentration of these components. The most severe skin damage was evoked by application of sodium lauryl sulfate. Noticeable skin damage was also seen with 2-methyl-1-butanol. No irritation to the skin resulted from treatment with isopropanol or sodium myristate. When test solution containing sodium lauryl sulfate was applied to the skin, a remarkable increment in fractal dimensions was noted. This may suggest that the structure of the skin was greatly compromised as a result of sodium lauryl sulfate application. Although no change in fractal dimension was observed as a result of application of the test solution containing only 25% isopropanol, the fractal dimension of skin cross section gradually increased with increasing concentrations of isopropanol, 2-methyl-1-butanol or sodium myristate in the test solutions. The increment of fractal dimension was around 0.4. Thus, the skin structure was also altered by the application of high concentrations of these compounds. Although the relevance to humans is not known, fractal analysis of skin structure is thought to be useful as a novel method for detecting skin damage that is brought about by the application of percutaneous absorption enhancers.

Animals↗
PubMed2000

A de novo philadelphia chromosome-positive acute mixed-lineage leukemia with both major and minor BCR/ABL mRNA transcripts.

A patient with a Philadelphia chromosome (Ph)-positive acute mixed-lineage leukemia (AMLL) expressing both major and minor BCR/ABL mRNA transcripts is described. Phenotypic analysis of the leukemic blasts revealed positivity for both myeloid and B-cell lineages. Southern blot analysis showed a rearrangement of the immunoglobulin heavy chain (IgH) gene. Reverse transcriptase polymerase chain reaction (RT-PCR) analysis showed the expression of both major and minor BCR/ABL mRNA transcripts. To our knowledge, this is the first report of AMLL expressing both major and minor BCR/ABL mRNA transcripts and rearrangement of the IgH gene.

Adult↗
PubMed2000

Enhanced enteral bioavailability of vancomycin using water-in-oil-in-water multiple emulsion incorporating highly purified unsaturated fatty acid.

The aim of this study was to evaluate the potential of an emulsion incorporating unsaturated fatty acids to improve the mucosal absorption of poorly absorbed drugs from rat intestinal loops in situ, using a water-in-oil-in-water (W/O/W) multiple emulsion. Vancomycin hydrochloride (VCM) was used as a model drug with low oral bioavailability. The entrapment efficiency of VCM in the emulsion was approximately 60% and remained constant over storage for 1 month at 4 degrees C. The emulsion incorporating C18 unsaturated fatty acids or docosahexaenoic acid (DHA) markedly enhanced VCM absorption after colonic and rectal dosing. The effectiveness of DHA on VCM colonic absorption improvement was the same as that of oleic acid, and less than that of linoleic and linolenic acids. For rectal dosing, bioavailability was similar among various emulsions, in the range 40-50%. The effect of the emulsion incorporating oleic acid or DHA on improving VCM enteral bioavailability was not increased proportional to the incorporated amount. The electrical resistance of membranes was not changed by the incorporation of various fatty acids in emulsions. Our results indicated that W/O/W emulsions incorporating C18 unsaturated fatty acid or DHA were useful carriers for improving the absorption of poorly absorbable drugs via the intestinal tract without gross changes to tight junction function.

Administration, Rectal↗
PubMed2000

A novel truncated isoform of the mannose-binding lectin-associated serine protease (MASP) from the common carp (Cyprinus carpio).

The mannose-binding lectin (MBL) and MBL-associated serine proteases (MASPs) play crucial roles in activation of the lectin pathway of the complement system. Mammals and Xenopus possess two distinct MASPs, MASP1 and MASP2, with different substrate specificity. Recently, a truncated form named MAp19 or sMAP, composed of N-terminal C1r/C1s/Uegf/bone morphogenetic protein (CUB)-1 and epidermal growth factor domains of MASP2, has been shown to be generated by alternative polyadenylation and splicing from the MASP2 gene. In the present study, we isolated cDNA encoding a novel MASP-related protein, designated MRP, from carp. MRP is distinct from MAp19/sMAP in containing two additional domains, CUB-2 and short consensus repeat (SCR)-1, followed by a unique C-terminal 21 amino acids, but resembles it by also lacking the serine protease domain, suggesting that carp MRP is a functional homologue of human MAp19/sMAP. Analyses of polymerase chain reaction (PCR)-amplified carp genomic DNA, from CUB-2 to SCR-2 of MASP, indicated that carp possess duplicated MASP genes, designated MASP-A and MASP-B, both of which contain an exon encoding the MRP-specific C-terminal stretch between the exons coding for SCR-1 and SCR-2 domains. Reverse transcription-PCR analysis showed that both MASP genes of carp produce the two MASP isoforms, MASP and MRP, through alternative polyadenylation and splicing. The conservation of MASP isoforms that lack the catalytic domain in both carp and human implies that they meet an essential requirement in the MBL-MASP complex of the lectin pathway.

Amino Acid Sequence↗
PubMed2000

KL-6 as a novel marker for activities of interstitial pneumonia in connective tissue diseases.

The objective of this study was to determine the role of serum KL-6 levels as a marker for the activity of interstitial pneumonia in patients with connective tissue diseases. The serum concentrations of KL-6, a glycoprotein produced mainly by pulmonary type II epithelial cells, were measured in 21 patients with connective tissue disease. The activity of interstitial pneumonia was compared with the associated serum KL-6 concentrations. Serum KL-6 concentrations in patients with interstitial pneumonia were significantly higher than those in the controls. Among patients with active interstitial pneumonia, serum KL-6 concentrations following the treatment (after improvement) were significantly lower than the pretreatment values. The extent of the pulmonary fibrosis correlated positively with the serum KL-6 concentrations during the inactive phase of the interstitial pneumonia. These results suggest that sequential measurement of serum KL-6 levels is a new and useful means for the evaluation of interstitial pneumonia in patients with connective tissue diseases.

Antigens↗
PubMed2000

Botryoid Wilms tumor: case report and review of literature.

A rare case of botryoid Wilms tumor is presented. The main clinical manifestations were persistent low-grade fever, malaise, and proteinuria associated with microhematuria. Ultrasonography revealed an echogenic mass in the right kidney, and a contrast-enhanced mass was found in the dilated collecting system by contrast-enhanced computed tomography. The surgically resected tumor was a polypoid, light-yellow, glistening mass that occupied a large part of the renal pelvis and originated from the pelvicaliceal wall. Part of the tumor extended to the proximal ureter, resulting in hydronephrosis in the involved kidney. No parenchymal lesion was observed. Microscopic examination revealed epithelial, stromal, and blastemal components, which indicated Wilms tumor. Infection had occurred in the hydronephrotic kidney, which presumably had caused the major presenting symptoms. The prognosis of our patient and previously reported cases of botryoid Wilms tumor was good compared with that of typical Wilms tumor, since the botryoid type can be detected at an early stage.

Hematuria↗
PubMed2000

Inhibition of L-leucine methyl ester mediated killing of THP-1, a human monocytic cell line, by a new anti-inflammatory drug, T614.

T614 (3-formylamino-7-methylsulfonylamino-6-phenoxy-4H-1-benzopyran-4-o ne) is a member of the family of methanesulfonanilide non-steroidal anti-inflammatory drugs (mNSAIDs), most of which act as cyclooxygenase (COX)-2 inhibitors. L-leucine methyl ester (Leu-OME) is a reagent which has been shown to kill phagocytes following interaction with intracellular proteases. There are two pathways whereby Leu-OME becomes cytotoxic to phagocytes. Within lysosomes, Leu-OME is converted into free Leu, which causes disruption of the lysosomes and subsequent cell necrosis. The other is the conversion of Leu-OME into (Leu-Leu)(n)-OME, which is associated with the induction of apoptosis. In the present study, we examined the action of T614 on Leu-OME mediated killing of THP-1, a human monocytic cell line. We revealed that T614 and phenylmethyl sulfonyl fluoride (PMSF), a serine protease inhibitor, inhibited Leu-OME mediated killing of THP-1 cells. All the other mNSAIDs, including nimesulide (NIM-03), fluosulide (CGP28238), FK3311 and NS398, also rescued THP-1 from Leu-OME mediated killing, although to a lesser degree. Of the classical NSAIDs tested, a protective effect was observed with diclofenac at high concentration, but not with naproxen or indomethacin. Unlike conventional lysosomal inhibitors, such as chloroquine and ammonium chloride (NH(4)Cl), T614 and PMSF did not raise lysosomal pH, as measured by flow cytometry using fluorescein isothiocyanate dextran (FITC-dextran). Therefore, the mechanism whereby T614 and PMSF inhibit Leu-OME killing is distinct from that of chloroquine or NH(4)Cl. Based on the similarity of T614 and PMSF, we suggest that, besides their roles as COX-2 inhibitors, T614 and other mNSAIDs may act as lysosomal protease inhibitors.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed2000

Neurosteroids ameliorate conditioned fear stress: an association with sigma receptors.

Mice exhibited a marked suppression of motility (conditioned fear stress) when placed in an environment in which they had previously received an electric footshock. This conditioned fear stress response was dose-dependently attenuated by neurosteroids such as dehydroepiandrosterone sulfate (DHEAS; 25 and 50 mg/kg, s.c.) and pregnenolone sulfate (PREGS; 10-50 mg/kg, s.c.), and by a putative sigma(1) receptor agonist, (+)-N-allylnormetazocine ((+)-SKF-10,047; 3 and 6 mg/kg, s.c.). However, progesterone (PROG; 10-50 mg/kg, s.c. ) and allopregnanolone (5 and 20 mg/kg, s.c.) had no effect on this stress response. The attenuating effects of DHEAS (50 mg/kg, s.c.), PREGS (50 mg/kg, s.c.), and (+)-SKF-10,047 (6 mg/kg, s.c.) were reversed by NE-100 (5 mg/kg, i.p.), a sigma(1) receptor antagonist and PROG (5 or 10 mg/kg, i.p.). When DHEAS (25 mg/kg) was co-administered with (+)-SKF-10,047 (3 mg/kg) at doses that do not affect the conditioned fear stress response by themselves, motor suppression was significantly attenuated. In mice showing the conditioned fear stress response, the serum concentration of DHEAS was lower than that in non-shocked mice. These results suggest that the attenuating effects of DHEAS and PREGS on the conditioned fear stress response are mediated via sigma(1) receptors and that PROG has a sigma(1) receptor antagonistic property. Further, the endogenous DHEAS may be involved in the expression of conditioned fear stress response in mice.

Animals↗
PubMed2000

Acid load during total parenteral nutrition: comparison of hydrochloric acid and acetic acid on plasma acid-base balance.

The effects of hydrochloric acid and acetic acid on the plasma acid-base balance were investigated in four rabbits receiving parenteral nutrition. Hyperchloremic metabolic acidosis was observed only in the animals receiving total parenteral nutrition (TPN) whose pH was adjusted with hydrochloric acid. The observed acidosis was due to an excess of hydrogen ions with chloride ions as judged by the plasma-base excess and urinary net-acid excess and not by the infusion of solution having a high titratable acidity. The hydrogen ion released from the acetic acid added to TPN is consumed by the metabolism of the acetate ion and thus does not contribute to the net hydrogen-ion load. A reduction in the chloride load by using acetic acid to adjust the pH of the TPN solution when it is formulated can be safely achieved and prevents acidosis.

Acetic Acid↗
PubMed2000

Pharmacokinetics and antitumor effect of doxorubicin carried by stealth and remote loading proliposome.

PURPOSE: The aim of the study was to prepare stealth and remote loading proliposome (SRP-L) to carry doxorubicin (DXR) and evaluate the pharmacokinetics, acute toxicity, and anticancer effect of DXR carried with SRP-L. METHODS: SRP-L was transparent solution. When SRP-L was injected into 0.9%, NaCl aqueous solution containing DXR. liposomes formed and automatically loaded DXR (SRP-L-DXR). The long circulation of SRP-L-DXR was evaluated using the pharmacokinetics of SRP-L-DXR. cardiolipin liposomal DXR (CL-DXR) and free DXR (F-DXR). The acute toxicity and anticancer effect of SRP-L-DXR were evaluated in C57BL/6 mice and murine hystocytoma M5076 tumor model. RESULTS: The average diameter of SRP-L-DXR in pure water was 112.9 +/- 8.6 (nm) and the encapsulation efficiency of SRP-L-DXR was 96.5 +/- 0.2% in pure water, 95.5 +/- 0.1% in 5% glucose and 98.01 +/- 0.6% in 0.9% NaCl. The plasma concentration of SRP-L-DXR was much higher than those of F-DXR and CL-DXR. Compared with that of F-DXR, the SRP-L-DXR had lower acute toxicity and its anticancer effects depended upon the therapeutic treatment. CONCLUSIONS: A novel proliposome (SRP-L) was developed, which could automatically load DXR and form SRP-L-DXR with excellent characteristics. SRP-L-DXR had lower acute toxicity but was not always more effective for the treatment of the ascitic M5076 than F-DXR.

Animals↗
PubMed2000

Effect of hyaluronan on monospermic penetration of porcine oocytes fertilized in vitro.

An experimental series was carried out to examine the ability of exogenous hyaluronan (HA) to facilitate monospermic penetration of in vitro-matured (IVM) oocytes during conventional porcine in vitro fertilization (IVF). IVM oocytes were co-cultured in fertilization medium with frozen-thawed, ejaculated boar spermatozoa. Addition of 0.5 mg/mL HA to the fertilization medium increased the rates of monospermic penetration (p < 0.05) on cumulus-denuded oocytes, without detrimentally affecting penetration rates (rates of monospermy were 57% and 78% in 2 mM caffeine-containing fertilization medium without and with HA, respectively). Caffeine and cumulus cells affected the rate of monospermic fertilization (p < 0.02-0.03), and caffeine affected the rate at which oocytes were penetrated (p < 0.02). Moreover, exogenous HA (0.5 mg/mL) in the sperm pre-incubation medium promoted monospermic penetration rates without affecting overall penetration rates. The effects of exogenous HA on sperm penetration and monospermy were also compared among semen batches. After semen was pooled from three fertile boars and held in homologous seminal plasma (SP) for 3 (batch A), 10 (batch B) or 20 (batch C) h prior to being cooled, spermatozoa from each semen batch were co-cultured with IVM oocytes. Monospermy of cumulus-denuded oocytes was increased (p < 0.05) by the addition of exogenous HA within the same semen batch (batches B and C). Furthermore, co-culture with cumulus cells did not mask the effects of HA on batch-B semen. The concentration of exogenous HA (0, 0.05 and 0.5 mg/mL) during sperm-oocyte co-culture differentially affected the rates of penetration and monospermy of cumulus-intact oocytes, depending on the time spermatozoa were held in SP before being frozen. The highest penetration rate was obtained by exposing semen to SP for 20 h prior to being cooled (0.05 mg/mL HA). The results suggest that exogenous HA decreases polyspermy during conventional porcine IVF. Furthermore, it appears that the action of HA is dependent upon the priming action of seminal plasma.

Animals↗
PubMed2000

Divergent structure of the ComQXPA quorum-sensing components: molecular basis of strain-specific communication mechanism in Bacillus subtilis.

In Bacillus subtilis, the ComQXPA quorum-sensing system controls cell density-dependent phenotypes such as the production of degradative enzymes and antibiotics and the development of genetic competence. Bacillus subtilis (natto) NAF12, a mutant defective in poly-gamma-glutamate (gamma-PGA) production, was derived from B. subtilis (natto) NAF4 by Tn917-LTV1 insertional mutagenesis. Determination of the mutant DNA sequences flanking the Tn917-LTV1 insert revealed that the insertion had inactivated comP in this mutant, indicating that gamma-PGA synthesis in B. subtilis (natto) is under the control of the ComP-ComA signal transduction system. A comparison of the amino acid sequences revealed striking variation in the primary structures of ComQ (44% identity), ComX (26%) and the sensor domain of ComP (36%) between B. subtilis (natto) NAF4 and B. subtilis 168. In contrast, the amino acid and nucleotide sequences of the kinase domains of ComP and of the ComA response regulator share 95% and 100% identity respectively. The comP genes of NAF4 and 168 restored the impaired competence of B. subtilis BD1658 (comP:cat) and gamma-PGA production of B. subtilis (natto) NAF12 (comP:Tn917-LTV1) to only 15% of the level achieved by the respective parent comP genes. However, when introduced together with the cognate comQ and comX genes, the comP genes restored the relevant defect of the heterologous comP mutants nearly to wild-type levels. Analogous to the comCDE system of Streptococcus strains and the agrBCDE system of Staphylococcus aureus, the concerted variation in the comQXP genes appears to establish specific intercellular communication between B. subtilis strains sharing the same pheromone system.

Amino Acid Sequence↗
PubMed2000

Genetic evidence for a novel gene(s) involved in urogenital development on 10q26.

BACKGROUND: Although the frequent association between distal 10q monosomy and urogenital anomalies suggests the presence of a gene(s) for urogenital development on distal 10q, molecular deletion mapping has not been performed for the putative gene(s). In this study, we examined genotype-phenotype correlations in patients with distal 10q monosomy. METHODS: This study consisted of six karyotypic males (cases 1 through 6) and four karyotypic females (cases 7 through 10) with 10q26 monosomy. Cases 3 through 5 and 7 through 10 had urinary anomalies such as vesicoureteral reflux and hypoplastic kidney, and cases 1 through 6, 8, and 9 exhibited genital anomalies such as micropenis, hypospadias, cryptorchidism, and hypoplastic labia majora. Fluorescence in situ hybridization (FISH) for 10q telomere, whole chromosome 10 painting, and microsatellite analysis for 35 loci on distal 10q were performed in cases 1 through 8. RESULTS: FISH and whole chromosome painting confirmed distal 10q monosomy in cases 1 through 8. Microsatellite analysis revealed that hemizygosity for the region distal to D10S186 was shared by cases with urinary anomalies and that for the region distal to D10S1248 was common to cases with genital anomalies. Furthermore, it was indicated that PAX2, GFRA1, and EMX2 on distal 10q, in which the deletions could affect urinary and/or genital development, were present in two copies in cases 1 through 8. CONCLUSIONS: The results suggest that a novel gene(s) for urinary development and that for genital development reside in the approximately 20 cM region distal to D10S186 and in the approximately 10 cM region distal to D10S1248, respectively, although it remains to be determined whether the two types of genes are identical or different.

Adolescent↗
PubMed2000

Transdermal absorption of L-dopa from a new system composed of two separate layers of L-dopa and hydrogel in rats.

To maintain the stability of L-dopa in hydrogel, a new system composed of two separate layers of L-dopa and hydrogel was developed. L-Dopa sheets were made by immersing L-dopa solution into wiper sheets and by lyophilizing them. Examination for stability of L-dopa in the L-dopa sheet revealed that its stability was maintained for at least 12 weeks, providing the sheet was kept at room temperature in a dark box. In a cutaneous absorption study of L-dopa in rats, an L-dopa sheet was attached to the shaved abdominal skin. A hydrogel composed of cutaneous absorption enhancers, water and ethanol, was spread on vinyl tape (hydrogel sheet), and this sheet was placed over the L-dopa sheet. L-Dopa that was administered transdermally effectively penetrated through the skin: The plasma level of L-dopa peaked at 30 min and remained high between 60 and 180 min after the cutaneous application. Our system, composed of two separated layers of L-dopa and hydrogel, enabled the stability of L-dopa to be maintained without losing transdermal absorption of L-dopa.

Administration, Cutaneous↗
PubMed2000

Interpretation and enantiomer analysis of methamphetamine abusers' urine and illegally brewed methamphetamine crystals.

This study deals with the high-performance liquid chromatographic identification of methamphetamine (MAMP) and amphetamine (AMP) enantiomers (d- and l-forms) in five illicit MAMP crystals and in urine specimens from 30 Japanese MAMP abusers. The analysis revealed that two of the types of crystals have a different optical purity ratio (l/d) and the other three have a single crystal of either the d- or l-enantiomer. The l/d ratios of two types of crystals were 0.04 and 49.4, and no racemic form (l/d = 1.00) was found. The urinary analysis showed that nonmetabolized MAMP and its demethylated metabolite, AMP, were present in urine specimens of all addicts. The stereoisomeric profiles in urine can be classified roughly into five groups according to the detected amount and the l/d ratio of MAMP and AMP enantiomers collected at one time point. In the first group, only d-MAMP and d-AMP were detected (in 16 cases). In the second group, only l-MAMP and l-AMP were detected (in one case). In the third group, the amount of the l-enantiomer detected, for both MAMP and AMP, was less than that of the d-enantiomer, and the l/d ratio was between 0.004 and 0.54 for MAMP and between 0.01 and 0.07 for AMP (in five cases). In the fourth group, the l-enantiomer of MAMP and AMP was found to be more abundant than the d-enantiomer, and the l/d ratio was between 2.63 and 30.11 for MAMP and between 1.23 and 31.30 for AMP (in four cases). In the fifth group, the amount of l-MAMP detected was greater than that of d-MAMP, and less l-AMP than d-AMP was detected. The l/d ratios were between 1.13 and 8.82 for MAMP and between 0.17 and 0.82 for AMP (in four cases). These results might be suitable for identification and the forensic toxicological investigation of AMP analogues.

Adolescent↗