Biomedical subjects

N Yokoyama

Publications and source records attributed to N Yokoyama.

At least 19 recordsLinked to original sources

PubMed2006

Amount of cholesterol in host membrane affects erythrocyte invasion and replication by Babesia bovis.

Cholesterol is a major component of the erythrocyte membrane. In the present study, we investigated the effects of cholesterol reduction in host bovine erythrocytes (RBC) on the growth of Babesia bovis, a major bovine haemoprotozoon. An in vitro growth assay with bovine RBC that had been prepared by pre-treatment with a cholesterol depletion agent (methyl-beta-cyclodextrin, MCD) showed that the culture with 5 mM MCD-treated RBC inhibited the growth of B. bovis significantly as compared with that with the control RBC. In further experiments, the treatment with 5 mM MCD was proved to suppress both activities of the parasite, erythrocyte invasion and replication within the infected RBC. In contrast, a slight reduction in the membrane cholesterol by 1 mM MCD treatment promoted both their growth and erythrocyte invasion activity. These results indicate that erythrocyte invasion and replication by B. bovis are affected by the amount of cholesterol in the host erythrocyte membrane.

Animals↗

PubMed2006

Number of positive lymph nodes independently affects long-term survival after resection in patients with ampullary carcinoma.

AIM: The nodal status is an established prognostic factor in ampullary carcinoma. The aim of this study was to compare the prognostic power of the anatomic location of positive nodes with that of the number of positive nodes. METHODS: Of 73 consecutive patients treated for ampullary carcinoma, 62 underwent pancreaticoduodenectomy with regional lymphadenectomy. A survival analysis of these 62 patients by nodal status was conducted retrospectively. A total of 1942 lymph nodes taken from the patients were examined histologically for metastasis. The location of positive regional nodes was classified into 4 categories, according to the Japanese staging system. The number of positive regional nodes was recorded for each patient. The median follow-up period was 124 months. RESULTS: Nodal disease was found in 31 patients, of whom 23 had 1-3 positive regional nodes and 8 had >or=4 positive regional nodes. Univariate analysis revealed that both the location (p<0.0001) and the number (p<0.0001) of positive nodes were significant prognostic factors. Multivariate analysis revealed that the number of positive nodes was an independent prognostic factor (p=0.007), while the location failed to remain as an independent variable. The median survival time was 59 months with a 5-year survival rate of 48% in patients with 1-3 positive nodes, whereas all patients with >or=4 positive nodes died of the disease within 29 months of resection (p=0.0001). CONCLUSION: The number, not the location, of positive regional lymph nodes independently affects long-term survival after resection in patients with ampullary carcinoma.

Adult↗

PubMed2006

The role of surface charging during the coadsorption of mercaptohexanol to DNA layers on gold: direct observation of desorption and layer reorientation.

We study the coadsorption of mercaptohexanol onto preimmobilized oligonucleotide layers on gold. Monitoring the position of the DNA relative to the surface by optical means directly shows the mercaptohexanol-induced desorption of DNA and the reorientation of surface-tethered strands in situ and in real time. By simultaneously recording the electrochemical electrode potential, we are able to demonstrate that changes in the layer conformation are predominantly of electrostatic origin and can be reversed by applying external bias to the substrate.

Adsorption↗

PubMed2006

Calcium-ions are involved in erythrocyte invasion by equine Babesia parasites.

Ethylene glycol bis (beta-aminoethylether)-N,N,N,N-tetraacetic acid (EGTA) is a chelating agent capable of binding to positively-charged metal ions, including a calcium-ion (Ca2+). Here, we demonstrated the inhibitory effect of the chemical on the in vitro asexual growth of the equine protozoan parasites, Babesia caballi and Babesia equi. The growth of both B. caballi and B. equi was significantly inhibited in the presence of EGTA (IC50=1.27 and 2.25 mM, respectively). Under microscopical observation, increased percentages of extracellular merozoites in the total parasites were detected in both of the cultures treated with high concentrations of EGTA. In contrast, further addition of Ca2+ to the EGTA-treated cultures prevented the parasites from clearing and the percentages of extracellular merozoites from increasing. As for B. caballi, an invasion test using high-voltage pulsing proved that EGTA has an inhibitory effect to their erythrocyte invasion. These results suggest that Ca2+ is involved in erythrocyte invasion by equine Babesia parasites.

Animals↗

PubMed2006

Effects of protein kinase inhibitors on the in vitro growth of Babesia bovis.

Staurosporine, Ro-31-7549, and KN-93, which are inhibitors of serine/threonine protein kinase, protein kinase C, and calcium-modulin kinase, respectively, were tested for their effects on the in vitro growth of Babesia bovis. Staurosporine was the most effective inhibitor, completely clearing the parasitaemia as early as the first day of exposure at a concentration of 100 microM. Moreover, staurosporine caused a significant increase in the percentage of extracellular merozoites, most likely due to the inhibition of erythrocyte invasion by the parasite. Although 5 mM Ro-31-7549 and KN-93 had a suppressive action, this was not enough to destroy the parasite. Interestingly, concentrations of 0.5 to 5 mM KN-93 influenced the parasitic development within the infected erythrocytes. The present study suggests that B. bovis requires, to a certain extent, the phosphorylations mediated by parasite- or host erythrocyte-protein kinases, in particular, for the processes of successful invasion of erythrocytes and intraerythrocytic development.

Animals↗

PubMed2005

Few-electron molecular states and their transitions in a single InAs quantum dot molecule.

We study electronic configurations in a single pair of vertically coupled self-assembled InAs quantum dots, holding just a few electrons. By comparing the experimental data of nonlinear single-electron transport spectra in a magnetic field with many-body calculations, we identify the spin and orbital configurations to confirm the formation of molecular states by filling both the quantum mechanically coupled symmetric and antisymmetric states. Filling of the antisymmetric states is less favored with increasing magnetic field, and this leads to various magnetic field induced transitions in the molecular states.

Journal Article↗

PubMed2005

Observation of the effects of radially sheared electric fields on the suppression of turbulent vortex structures and the associated transverse loss in GAMMA 10.

Vortexlike turbulent structures in hot-ion mode plasmas with several keV are observed in the case with a radially produced weak shear of electric fields E(r). However, a strong E(r) shear formation due to a high ion-confining potential phi(c) production clears up these vortices together with plasma-confinement improvement and disappearance of both drift-wave and turbulencelike Fourier spectral signals. These findings are based on three-time progress in phi(c) in comparison to phi(c) attained 1992-2002. The significant advance of phi(c) is well extended in line with proposed potential-formation physics scalings.

Journal Article↗

PubMed2005

Clinical significance of Ki-67 proliferation index in disease progression and prognosis of patients with resected colorectal carcinoma.

BACKGROUND: The prognostic role of proliferation markers in patients with colorectal neoplasia is inconclusive. The aim of this study was to evaluate proliferation markers in patients with colorectal carcinoma, to relate these to standard clinicopathological findings and to investigate their potential role in the prediction of long-term survival. METHODS: Archived formalin-fixed, paraffin-embedded tissue samples from a cohort of 106 patients who underwent curative resection for colorectal carcinoma were analysed to determine a Ki-67 (MIB-1)-derived proliferation index (PI) using a double immunostaining technique. The relationship between PI and standard clinicopathological variables was assessed and its association with long-term survival evaluated. RESULTS: A significant association was found between PI and tumour (T) status (P = 0.001), lymph node (N) involvement (P = 0.0098), the presence of distant metastases (P < 0.010) and advanced stage of disease (P < 0.001). On multivariate analysis, PI was shown to be an independent prognostic factor for long-term survival (hazard ratio 2.1 (95 per cent confidence interval 1.1 to 4.1); P = 0.032). CONCLUSION: Cell proliferation is significantly associated with tumour progression and may be used to identify patients with a predicted adverse outcome after resection of colorectal carcinoma.

Adult↗

PubMed2005

Host serum modifies the drug susceptibility of Babesia bovis in vitro.

Babesia parasites generally require a defined percentage of serum in the culture medium for their in vitro growth. In this study, we attempted to culture Babesia bovis in a serum-free condition. The growth pattern and morphology of B. bovis in serum-free (plain) GIT medium were unaltered as compared to those of the standard growth condition containing 40% bovine serum in M199. When exposed to the test drugs, the parasite in plain GIT medium showed clearly lower IC50 values than those in 40% serum-containing G IT medium, indicating that several serum components may interfere with the drug bio-availability. Therefore, the serum-free culture system is useful for standardizing drug test protocols and understanding the roles of serum factors in the drug test.

Animals↗

PubMed2005

Molecular characterization of a putative protein disulfide isomerase from Babesia caballi.

We produced a mAb against the Babesia caballi extracellular merozoite termed mAb 2H2 and used it to screen a cDNA expression library prepared from B. caballi merozoite mRNA for highly expressed proteins. The complete nucleotide sequence of the cloned gene had 1547 nucleotides and contained a 36-nucleotide intron. The 1398 nucleotide open reading frame predicts a 51 kDa protein showing similarity to protein disulfide isomerase (PDI) from other species. The PDI gene had a predicted N-terminal signal sequence of 19 amino acids and a C-terminal tetrapeptide sequence (His-Thr-Glu-Leu; HTEL) for retention in lumen of the endoplasmic reticulum (ER). The recombinant protein expressed in baculovirus showed an apparent mass of 51 kDa, identical to that the native B. caballi protein. Moreover, the ER retention signal site (HTEL) of the recombinant protein retained its function in ER of insect cells. This 51 kDa protein was strongly expressed by extracelluar B. caballi merozoites in indirect immunofluorescence antibody tests, and was not expressed in the early phase of trophozoite development. Interestingly, detailed observation showed that the reaction of anti-P51 antibody and mAb 2H2 against pear-shaped forms was very erratic, some displaying one or two brightly fluorescent patterns.

Amino Acid Sequence↗

PubMed2005

Left ventricular myocardial mass determination by contrast enhanced colour Doppler compared with magnetic resonance imaging.

OBJECTIVE: To assess the feasibility of using contrast enhanced colour Doppler echocardiography to determine left ventricular (LV) mass and to compare its accuracy with LV mass obtained by magnetic resonance imaging (MRI). METHODS: Images were acquired in the short axis plane of the heart, derived from coronal and sagittal scout views and double oblique angulation. The LV mass was calculated by two methods: Simpson's rule and the area-length method. Levovist (Schering AG, Berlin, Germany) 2.5 g was given by slow intravenous bolus or infusion over about 45 seconds for contrast imaging. LV images were captured in the apical two chamber, four chamber, and three chamber views. Each contrast harmonic colour Doppler image was converted to a cavity-only image by simple image mathematics. RESULTS: 27 (77.1%) of the patients (mean (SD) age 66.2 (8.9) years) were men. There was a mean (SD) interval of 6.6 (8.6) days (range 0-27 days) between echocardiography and MRI. The mean (SD) LV mass determined by MRI Simpson's rule method was 171.0 (52.4) g (range 105.1-318.7 g). The mean LV mass (SD) determined by the echocardiographic Simpson's rule method was 178.2 (47.0) g (range 112.6-307.6 g). The mean (SD) MRI area-length LV mass was 187.3 (64.5) g (range 109.0-393.6 g). The linear regression correlation between LV mass determined by MRI Simpson's and echocardiographic Simpson's methods was excellent (y = 1.022x, R2 = 0.986) with a mean (SD) difference of 7.20 (20.9) g. The linear regression correlation between the MRI area-length LV mass and MRI Simpson's LV mass was excellent (y = 1.101x, R2 = 0.989) with a mean (SD) difference of 16.3 (22.3) g. CONCLUSIONS: LV mass may be obtained reliably by contrast enhanced colour Doppler and two dimensional echocardiography. The contrast Doppler method accurately determines LV mass with excellent agreement with the MRI technique.

Aged↗

PubMed2004

Single-dot spectroscopy via elastic single-electron tunneling through a pair of coupled quantum dots.

We study the electronic structure of a single self-assembled InAs quantum dot by probing elastic single-electron tunneling through a single pair of weakly coupled dots. In the region below pinch-off voltage, the nonlinear threshold voltage behavior provides electronic addition energies exactly as the linear, Coulomb blockade oscillation does. By analyzing it, we identify the s and the p shell addition spectrum for up to six electrons in the single InAs dot, i.e., one of the coupled dots. The evolution of the shell addition spectrum with magnetic field provides Fock-Darwin spectra of the s and p shells.

Journal Article↗

PubMed2004

High risk of gallbladder carcinoma in elderly patients with segmental adenomyomatosis of the gallbladder.

The clinical significance of adenomyomatosis of the gallbladder remains unclear. This study aimed to clarify the relationship between segmental adenomyomatosis and gallbladder carcinoma, and to elucidate the histogenesis of gallbladder carcinoma associated with segmental adenomyomatosis. A total of 4,560 consecutive patients underwent cholecystectomy. The specimens were examined grossly and histologically. Adenomyomatosis of the gallbladder was divided into segmental, fundal, and diffuse types. Sixty noncancerous gallbladders with segmental adenomyomatosis were examined for epithelial metaplasia. The incidence of gallbladder carcinoma was higher in patients with segmental adenomyomatosis (22/334, 6.6%) than in those without (181/4226, 4.3%; P=0.049). This difference was more marked among patients equal to or older than 60 years of age (15/96,15.6% versus 147/2407, 6.1%, respectively; P<0.001). The other types of adenomyomatosis did not show any significant increases in the incidence of gallbladder carcinoma. In all 22 patients with both segmental adenomyomatosis and carcinoma, the tumors developed only in the fundal mucosa. Epithelial metaplasia was more marked in the fundal mucosa of segmental adenomyomatosis than in the neck mucosa (P=0.003). Segmental adenomyomatosis is a high-risk condition for gallbladder carcinoma, especially in elderly patients. Epithelial metaplasia appears to be related to increased carcinogenesis in the fundal mucosa of segmental adenomyomatosis.

Adenomyoma↗

PubMed2003

Morphological studies on the extracellular structure of the midgut of a tick, Haemaphysalis longicornis (Acari: Ixodidae).

In the present study, morphological observations on the extracellular structures found on the apical surface of the midgut epithelium, known as the peritrophic membrane (PM) or glycocalyx, are described in Haemaphysalis longicornis females and larvae. These structures have been hypothesized to provide protection to the microvilli of epithelial cells of the digestive tract. Our aim was to determine whether the extracellular structures are important in the digestion of the blood meal and/or as a protection against infection or injury. The PM was detectable in the midgut of engorged larvae by electron microscopy, but not in engorged females. However, a PM-like structure, stainable with toluidine blue, was observed in females by light microscopy. From the results of confocal laser scanning and electron microscopic observations with wheat germ agglutinin (WGA lectin) staining for chitin of the PM, however, the structure was clearly recognized. The structure in the female is likely to be PM because staining with WGA lectin in the presence of GlcNAc indicates the presence of chitin and various morphologies of PM have been reported in insects and ticks. These results show morphologically that different types of PM-like structure are formed in larvae and females of H. longicornis.

Animals↗

PubMed2003

Clotrimazole, ketoconazole, and clodinafop-propargyl inhibit the in vitro growth of Babesia bigemina and Babesia bovis (Phylum Apicomplexa).

We evaluated the growth inhibitory efficacy of the imidazole derivatives, clotrimazole (CLT) and ketoconazole (KC), and the herbicide clodinafop-propargyl (CP), in in vitro cultures of Babesia bovis and B. bigemina. Clotrimazole was effective in a dose range of 15 to 60 microM (IC50: 11 and 23.5 microM), followed by KC (50 to 100 microM; IC50: 50 and 32 microM) and CP (500 microM; IC50: 265 and 390 microM). In transmission electron microscopy, extensive damage was observed in the cytoplasm of drug-treated parasites. Combinations of CLT/KC, CLT/CP and CLT/KC/CP acted synergistically in both parasites. In contrast, the combination of KC/CP was exclusively effective in B. bovis, but not in B. bigemina.

Animals↗

PubMed2003

Hepatitis viral status affects the pattern of intrahepatic recurrence after resection for hepatocellular carcinoma.

AIM: To define whether the patterns of intrahepatic recurrence after resection for hepatocellular carcinoma differ according to hepatitis viral status. METHODS: One hundred and eleven patients undergoing a curative resection for hepatocellular carcinoma were divided into three groups: the C-viral group (n=55), which tested positive for hepatitis C antibody; the B-viral group (n=32), which tested positive for hepatitis B surface antigen; and the non-B non-C (NBNC) group (n=24), which tested negative for both hepatitis B surface antigen and hepatitis C antibody. The long-term outcomes were analyzed retrospectively. RESULTS: The pattern of development of intrahepatic recurrence differed between the NBNC group and the other groups: the cumulative probability of intrahepatic recurrence reached a plateau at 2.4 years after resection in the NBNC group, while it continued to increase steadily in the hepatitis viral groups. The C-viral group showed a higher incidence of intrahepatic recurrence than the other groups by univariate (P=0.0306) and multivariate (relative risk=1.69, P=0.0429) analyses. Multiple intrahepatic recurrent lesions were more common in the C-viral group (P=0.0457). CONCLUSIONS: Multicentric carcinogenesis in the remnant liver was less common in the NBNC group than in hepatitis viral groups. Hepatitis C virus infection is a significant risk factor for intrahepatic recurrence after resection and is also associated with multiple intrahepatic recurrent lesions.

Adolescent↗

PubMed2003

Ultrastructural studies on the pheromone-producing cells in the silkmoth, Bombyx mori: formation of cytoplasmic lipid droplets before adult eclosion.

In Bombyx mori, pheromone-producing cells accumulate a number of lipid droplets in the cytoplasm preceding the production of the sex pheromone, bombykol. The process of lipid droplet formation in the pheromone-producing cells was investigated by using light and electron microscopy. Light microscopy revealed that the lipid droplets appeared from 2 days before adult eclosion and dramatic accumulation took place between 2 days and 1 day before eclosion. Electron microscopical studies revealed that smooth endoplasmic reticulum and numerous vesicles, their sizes being less than 1 microm, were detectable 2 days before eclosion, and some vesicles were fused with mitochondria at this stage. These characteristic changes in the pheromone-producing cells suggest that fatty acyl-CoA synthesis following de novo fatty acid synthesis takes place at this time. Involutions in the basal plasma membrane of the cells occurred throughout the observed period, which were extensive on the day before adult eclosion. Besides extensive basal involutions, immature lipid droplets appeared and then mature fully electron-dense lipid droplets were observed on the day of adult eclosion. These ultrastructural observations, combined with recent physiological studies suggest, that the basal involutions presumably reflect the uptake of lipidic components required for the construction of lipid droplets, the function of which is to store the bombykol precursor and to provide it for bombykol biosynthesis in response to pheromonotropic stimuli by pheromone biosynthesis activating neuropeptide (PBAN).

Animals↗

PubMed2002

cDNA cloning of calcineurin heterosubunits from the pheromone gland of the silkmoth, Bombyx mori.

Pheromone biosynthesis activating neuropeptide (PBAN) stimulates the step of fatty acyl reduction in the pheromone biosynthetic pathway of the silkmoth, Bombyx mori. It has been suggested that the intracellular signal transduction of PBAN in B. mori involves Ca(2+), calmodulin, and calcineurin (also known as protein phosphatase 2B). We have cloned two cDNAs encoding calcineurin heterosubunits from a pheromone gland cDNA library of B. mori. The 2,996-bp clone predicts a 495-amino acid protein homologous to the catalytic subunit calcineurin A (CnA) with a molecular mass of 55,968. The deduced amino acid sequence well conserves the calcineurin B (CnB)-binding domain and two subdomains, a calmodulin-binding and an autoinhibitory domain, showing 77-85% and 82% identities to the isoforms of Drosophila melanogaster CnA and human CnA, respectively. On the other hand, the 820-bp clone predicts a 170-amino acid protein homologous to the regulatory subunit CnB with a molecular mass of 19,357. The deduced amino acid sequence well conserves four EF-hand type calcium-binding structures, showing 95% and about 85% identities to D. melanogaster CnB and mammalian CnBs, respectively. A yeast two-hybrid system has demonstrated the molecular interaction between B. mori CnA and CnB. Northern blot analyses revealed that both CnA and CnB genes were expressed in various larval and adult tissues of B. mori. Both transcripts detected in the pheromone gland three days before adult eclosion increased by the day before eclosion and the mRNA levels were found to be high even two days after adult eclosion. Immunohistochemical analysis has revealed that B. mori calcineurin is localized in the cytoplasm of the pheromone-producing cells.

Amino Acid Sequence↗