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Biomedical subjects

N Kurita

Publications and source records attributed to N Kurita.

At least 19 recordsLinked to original sources

Multiple organ damage caused by a new toxin azaspiracid, isolated from mussels produced in Ireland.

A new type of food poisoning resulting from ingestion of mussels produced in Ireland occurred in the Netherlands in 1995 and then reoccurred in Ireland in 1997. As the causative agent, azaspiracid, was isolated in pure form and revealed to have a structure entirely unlike other known algal toxins, in vivo studies with mice were carried out to elucidate the pathological injuries caused by the toxin. By per os administration, the toxin caused necrosis in the lamina propria of the small intestine and in lymphoid tissues such as thymus, spleen and the Peyer's patches. Both T and B lymphocytes were injured. Additionally a fatty change was observed in the liver. These injuries distinctly differed from those caused by the representative diarrhetic shellfish toxin, okadaic acid.

Administration, Oral↗

Density functional MO calculation for stacked DNA base-pairs with backbones.

In order to elucidate the effect of the sugar and phosphate backbones on the stable structure and electronic properties of stacked DNA base-pairs, we performed ab initio molecular orbital (MO) calculations based on the density functional theory and Slater-type basis set. As a model cluster for stacked base-pairs, we employed three isomers for the dimer unit of stacked guanine-cytosine pairs composed with backbones as well as base-pairs. These structures were fully optimized and their electronic properties were self-consistently investigated. By including the backbones, the difference in total energy among the isomers was largely enhanced, while the trend in relative stability was not changed. The effect of backbones on the electronic properties is remarkable: the MOs with the character of the PO4 parts of backbones appear just below the highest-occupied MO. This result indicates that the PO4 parts might play a rule as a reaction site in chemical processes concerning DNA. Therefore, we conclude that the DNA backbones are indispensable for investigating the stability and electronic properties of the stacked DNA base-pairs.

Base Pairing↗

Effect of cytokines on antifungal activity of human polymorphonuclear leucocytes against yeast cells of Paracoccidioides brasiliensis.

In our previous study, it was observed that human peripheral blood polymorphonuclear leucocytes (PMNs) exhibited a fungistatic effect on yeast cells of Paracoccidioides brasiliensis, and that interferon-gamma (IFN-gamma), but not tumor necrosis factor-alpha or interleukin-8 (IL-8), enhanced the antifungal activity of PMNs. In the present study, granulocyte-macrophage colony-stimulating factor (GM-CSF) also enhanced the PMN activity. GM-CSF-activated PMNs exhibited a killing effect on P. brasiliensis isolate Bt-4 and an enhanced fungistatic effect on isolate Aoki. IL-1beta activated PMNs to kill isolate Bt-4. Granulocyte colony-stimulating factor had no effect. Combinations of IFN-gamma with GM-CSF or IL-1beta, but not a combination of GM-CSF and IL-1beta, exhibited a synergistic effect in enhancing the antifungal activity of PMNs. These results strongly suggest that PMNs activated with IFN-gamma, GM-CSF and/or IL-1beta might play an important role in host defense in early infection with P. brasiliensis due to their enhanced antifungal activity.

Colony Count, Microbial↗

Fungistatic and fungicidal activities of murine polymorphonuclear leucocytes against yeast cells of Paracoccidioides brasiliensis.

Recently, a novel culture medium for detecting live yeast cells of Paracoccidioides brasiliensis was developed by Kurita et al. Using this culture medium, murine peritoneal polymorphonuclear leucocytes (PMN) were examined for fungistatic and fungicidal activities against P. brasiliensis yeast cells. The magnitude of the antifungal effect of PMN varied depending upon the fungal isolates used. PMN exhibited a killing effect on P. brasiliensis isolate Bt-4 in 2 h of coculture. In contrast, the other three fungal isolates employed were resistant to killing by PMN. However, PMN considerably suppressed the growth of isolates Tatu and Recife in a long-term assay (approximately 72 h). The growth of isolate Bt-9 was also suppressed by PMN during the first 24 h, but was found to be considerably promoted at 72 h of coculture. Interferon-gamma (IFN-gamma), but not tumour-necrosis factor-alpha, significantly augmented the antifungal activity of PMN. IFN-gamma-treated PMN exhibited a killing effect on isolates Tatu, Recife and Bt-9 after 24 h of coculture, and showed an enhanced killing effect on isolate Bt-4. Contact between PMN and fungal cells was required for PMN to exert the antifungal effect. Our results suggest that PMN, whether activated with cytokines or not, might play a critical role in host resistance in early infection with this fungus by buying time for development of more effective immunologic responses.

Animals↗

Antifungal activity of human polymorphonuclear leucocytes against yeast cells of Paracoccidioides brasiliensis.

Human peripheral blood polymorphonuclear leucocytes (PMN) were examined in vitro for antifungal activity against yeast cells of Paracoccidioides brasiliensis. The yeast cell of this fungus was resistant to killing by PMN. However, PMN exhibited a fungistatic effect on the fungal isolates employed in a long-term ( approximately 72 h) assay. Lysates of PMN did not show a fungistatic or fungicidal effect, indicating that live PMN are necessary for the antifungal effect. Interferon-Gamma (IFN-Gamma) enhanced the antifungal activity of PMN. IFN-Gamma-treated PMN killed isolate Bt-4 within 2 h of coculture, and after 24 h a still greater killing effect was observed. By contrast, IFN-Gamma-treated PMN did not show a significant killing effect on isolate Tatu, but did exhibit an enhanced fungistatic effect on this isolate. In contrast, tumor necrosis factor-alpha and interleukin 8 had no effect on the antifungal activity of PMN. Scanning electron microscopy revealed that the surface structure of the fungal cell was apparently damaged by PMN within 24 h of cocultures. Based on these novel findings, we speculate that human PMN might play a role in host resistance in early infection with this fungus due to their antifungal activity.

Cell Division↗

The effect of methyl linoleate hydroperoxides and radical initiator 2,2'-azobis (2-amidinopropane) dihydrochloride on antibody production by mouse spleen cells.

Methyl linoleate hydroperoxides (MLHPO) and 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH), a widely used free radical initiator, were examined for effects on antibody production by spleen cells using plaque-forming cell response against sheep red blood cells (SRBC). The in vitro primary antibody response was enhanced in the presence of MLHPO at a concentration range of 2-20 microM or AAPH at 0.1 microM, but was suppressed with higher concentrations of these compounds. In the in vitro secondary antibody responses, both MLHPO and AAPH failed to increase plaque-forming cell response above that of the control culture. Following the oral administration of MLHPO (2.29 g/kg) four times to mice, in vivo primary plaque-forming cell response was significantly suppressed. After, a single intraperitoneal injection of AAPH (60 mg/kg) to mice, in vitro primary plaque-forming cell response was also suppressed. These findings suggest that primary antibody response can be affected by lipid hydroperoxides and oxygen radicals in vivo.

Amidines↗

A comparative study of four different staining methods for estimation of live yeast form cells of Paracoccidioides brasiliensis.

A comparative study of four different staining methods for estimation of live yeast form cells of Paracoccidioides brasiliensis was carried out. The staining methods used were fluorescent staining, vital dye exclusion tests with erythrosin B and by Janus green and lactophenol cotton blue staining. Colony forming units (cfu) of the yeast form of eight P. brasiliensis isolates on brain heart infusion agar (BHIA) supplemented with 4% horse serum plus 5% P. brasiliensis cell extract (BHIA + HS + EXT) were examined for reliability of staining in determining the number of live fungal units in eight different isolates. Cfu on BHIA + HS + EXT plates showed an excellent plating efficiency over 96% in all isolates tested. The percentage of the live cells indicated by fluorescent staining (FL) or vital dye exclusion test with erythrosin B (EB) or Janus green (JG-1) was lower than that of cfu. By contrast, the percentage due to modified dye exclusion test with Janus green (JG-2) and that due to lactophenol cotton blue staining (LPCB) showed a close correration to that of cfu. Our results indicate that the modified dye exclusion test with Janus green and lactophenol cotton blue staining are useful for estimating cell viability of yeast form cells of P. brasiliensis.

Azo Compounds↗

An improved culture medium for detecting live yeast phase cells of Paracoccidioides brasiliensis.

The plating efficiency of standard mycological media such as brain heart infusion (BHI) agar is poor for Paracoccidioides brasiliensis. We prepared a water-extract of yeast phase cells of P. brasiliensis and examined it for growth-enhancing activity for the fungus. The water-extract, when added to BHI agar to a concentration of 5%, improved the plating efficiency of the medium for the fungus to some extent, but the degree of improvement was considerably varied among P. brasiliensis isolates. By contrast, when the water-extract was added in combination with horse serum (4%), the plating efficiency was highly improved (to 94-99%) for all the P. brasiliensis isolates employed. The growth-enhancing factor(s) in the water-extract was heat-stable and heating at 120 degrees C for 15 min had little, if any, effect on growth-enhancing activity.

Culture Media↗

Comparative studies on the postantifungal effect produced by the synergistic interaction of flucytosine and amphotericin B on Candida albicans.

A turbidometric method was used to measure Candida albicans yeast cell growth and to quantitate the postantifungal effect (PAFE) after exposure to various concentrations of flucytosine and amphotericin B, alone and in combination, for 2 hr at 30 degrees C. The drug concentrations used in the PAFE assays were determined by initial MIC and FIC (fractional inhibitory concentration) evaluations. The PAFE was calculated by the difference in time (hr) required for growth of the control and test cultures to reach the 0.5 absorbance level following removal of the drug by dilution. A synergistic PAFE was evidenced with combinations of the two drugs at concentrations below their individual MICs. Combinations of flucytosine (0.012 to 0.049 micrograms ml-1) and amphotericin B (0.195 to 0.39 micrograms ml-1) produced PAFEs ranging from 6.3 to 21.8 hr. These PAFEs persisted from 0.3 to 14.7 hr longer than those achieved when each of the two agents was assayed separately.

Amphotericin B↗

An early effect of aflatoxin B1 administered in vivo on the growth of bone marrow CFU-GM and the production of some cytokines in rats.

Our data demonstrate the granulopoietic toxicity of aflatoxin B1 (AFB1) in vivo and show an impact of this mycotoxin on the production of some humoral regulatory factors dealing with the granulopoietic developmental pathway (CSA, IL-1, IL-2). The dose of AFB1 studied represented approximately 1/5 of LD50 for young male rats. An early suppressive effect of AFB1 towards CFU-GM was transient in treated animals. The peak in granulopoietic activity was preceded in time by an increased CSA and IL-1 formation. Elevated IL-2 synthesis and increased T cell activation paralleled the peak in granulopoietic activity.

Aflatoxin B1↗