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Biomedical subjects

E Nagao

Publications and source records attributed to E Nagao.

At least 19 recordsLinked to original sources

Risk evaluation for staphylococcal food poisoning in processed milk produced with skim milk powder.

The growth of S. aureus and the production of staphylococcal enterotoxin A (SEA) in skim milk concentrates stored at inappropriate temperatures in a recovery milk tank (tank for excess concentrated skim milk) used in the manufacture of skimmed milk powder were investigated. Also, it was estimated if a possible outbreak of food poisoning would occur if the contaminated skimmed milk powder was used in the manufacture of processed milk. Skim milk concentrates with milk solid content of 15, 25, and 35% were inoculated with S. aureus at 1-2 log CFU/ml and incubated at 15, 25, or 35 degrees C for 0 to 24 h with or without shaking. Bacterial growth and the level of SEA production were measured. At 35 degrees C with shaking, there was a significant difference (p<0.05) in one way layout analysis of variance, and it was demonstrated that the growth of S. aureus and SEA production could be milk solid content-dependent. Shaking accelerated the growth of S. aureus and SEA production at 35 degrees C. Generally, skim milk powder is produced by mixing a set percentage of skim milk concentrates (recovery milk) from the recovery milk tank into raw milk. If recovery milk contaminated with S. aureus at levels of 1-2 log CFU/ml is kept at 15 to 35 degrees C due to a power failure, it was estimated that processed milk consumption of 670-1200 ml, 420-1500 ml and 18-83 ml would trigger the onset of food poisoning symptoms when skim milk concentrates (recovery milk) are stored at 25 degrees C for 24 h, 35 degrees C for 10 h, and 35 degrees C for 24 h, respectively, during the production of the skim milk powder. Based on these consumption levels, it was concluded that, if recovery milk cannot be refrigerated and is stored at room temperature (25 to 35 degrees C), it must be used within 8 h and preferably within 6 h.

Animals↗

Comparison between ultrafiltration and trichloroacetic acid precipitation method for concentration of Staphylococcus aureus enterotoxin in dairy samples.

Staphylococcus aureus enterotoxin that may be contained at low concentrations in milk and dairy products can cause food poisoning. To detect this enterotoxin at low concentrations, samples should be concentrated. We evaluated the performance of centrifugal ultrafiltration method (UF) in comparison with trichloroacetic acid precipitation method (TCA) for the concentration of S. aureus enterotoxin in milk and dairy products. S. aureus enterotoxin A (SEA) were added at various concentrations to ultra high-temperature heating process (UHT) milk, UHT concentrated skim milk, UHT skim milk powder, low heat-treated (LH) skim milk powder, and raw milk. SEA was concentrated by TCA and UF once a day on a total of 3 days by different researchers to prepare test solutions. The fluorescence value (TV) of test solutions was determined using an immunofluorescence autoanalyzer (miniVIDAS), and the linearity and slope of the regression line, relative standard deviation (RSD(RW)) at each added concentration, detection limit (DL), quantification limit (QL), and the recovery rate by each concentration method were obtained according to the guidelines of the International Conference on Harmonization (ICH). The slope of the regression line obtained by UF was steeper than that by TCA for all dairy samples excluding LH (74 degrees C, 20 s) skim milk powder. RSDRW, DL, and QL obtained by UF were comparable to or more excellent than those obtained by TCA. The procedure of UF was simpler than that of TCA. The recovery rate and rapidity were similar between the two methods. The DL and QL of enterotoxins other than SEA in dairy products by UF or TCA were estimated based on the DL and QL of SEA. In this estimation, consideration was given to reactions between each enterotoxin and its antibody, and also to the immunoactivity maintenance rate of each enterotoxin after addition of trichloroacetic acid in TCA. The estimated values were similar to those obtained by experiments using enterotoxin C1 (SEC1). UF using a centrifugal ultrafiltration membrane can be more readily performed and similar to or more reliable than TCA. UF combined with a miniVIDAS can be used for quantitative routine analysis.

Animals↗

Gluteal muscular and sciatic nerve metastases in advanced urinary bladder carcinoma: case report.

We present a case of gluteal muscular and sciatic nerve metastases from urinary bladder carcinoma. T2-weighted magnetic resonance images demonstrated diffuse swelling and an increase in the signal of the right gluteus maximus muscle without destruction of the original arrangement of muscular fibers. Further, remarkable thickening of the right sciatic nerve showing a relatively hypointense signal was detected. Postcontrast T1-weighted images showed strong enhancement of these structures. Fine-needle aspiration biopsy with ultrasonographic guidance confirmed metastatic carcinoma cells in the right gluteal muscle and the sciatic nerve. These radiologic findings may represent a rare pattern of metastasis from urinary bladder carcinoma.

Adenocarcinoma↗

Rapid determination of bovine lactoferrin in dairy products by an automated quantitative agglutination assay based on latex beads coated with F(ab')2 fragments.

UNLABELLED: This study evaluated an automated immunoassay for bovine lactoferrin (LF) in dairy products based on latex beads coated with F(ab')2 fragments. METHODS: F(ab')2 fragments were obtained by pepsin digestion of rabbit anti-bovine LF (IgG fraction) and polystyrene latex beads were coated with the F(ab')2 fragments. We used the beads to develop a rapid and homogeneous light scatter immunoassay employing an autoanalyzer (the Automated Latex assay). The Automated Latex assay was easy to perform and could rapidly determine bovine lactoferrin in lactoferrin-supplemented products. It was sensitive enough for testing products and showed good precision.

Animals↗

Differences in recombination frequencies during female and male meioses of the sex chromosomes of the medaka, Oryzias latipes.

In the medaka, Oryzias latipes, sex is determined chromosomally. The sex chromosomes differ from those of mammals in that the X and Y chromosomes are highly homologous. Using backcross panels for linkage analysis, we mapped 21 sequence tagged site (STS) markers on the sex chromosomes (linkage group 1). The genetic map of the sex chromosome was established using male and female meioses. The genetic length of the sex chromosome was shorter in male than in female meioses. The region where male recombination is suppressed is the region close to the sex-determining gene y, while female recombination was suppressed in both the telomeric regions. The restriction in recombination does not occur uniformly on the sex chromosome, as the genetic map distances of the markers are not proportional in male and female recombination. Thus, this observation seems to support the hypothesis that the heterogeneous sex chromosomes were derived from suppression of recombination between autosomal chromosomes. In two of the markers, Yc-2 and Casp6, which were expressed sequence-tagged (EST) sites, polymorphisms of both X and Y chromosomes were detected. The alleles of the X and Y chromosomes were also detected in O. curvinotus, a species related to the medaka. These markers could be used for genotyping the sex chromosomes in the medaka and other species, and could be used in other studies on sex chromosomes.

Animals↗

Plasmodium falciparum-infected erythrocytes: qualitative and quantitative analyses of parasite-induced knobs by atomic force microscopy.

We used the combination of an atomic force microscope and a light microscope equipped with epifluorescence to serially image Plasmodium falciparum-infected erythrocytes. This procedure allowed us to determine unambiguously the presence and developmental stage of the malaria parasite as well as the number and size of knobs in singly, doubly, and triply infected erythrocytes. Knobs are not present during the ring stage of a malaria infection but a lesion resulting from invasion by a merozoite is clearly visible on the erythrocyte surface. This lesion is visible into the late trophozoite stage of infection. Knobs begin to form during the early trophozoite stage of infection and have a single-unit structure. Our data suggest the possibility that a two-unit structure of knobs, which was reported by Aikawa et al. (1996, Exp. Parasitol. 84, 339-343) using atomic force microscopy, appears to be a double-tipped image. The number of knobs per unit of host cell surface area is directly proportional to parasite number in both early and late trophozoite stages. These results indicate that knob formation by one parasite does not influence knob formation by other parasites in a multiply infected erythrocyte. In addition, knob volume is not influenced by either parasite stage or number at the late trophozoite stage, indicating that the number of component molecules per knob is constant throughout the parasite maturation process.

Animals↗

Two distinct hemolysins in Trichomonas tenax ATCC 30207.

An oral protist Trichomonas tenax ATCC 30207 was investigated for the ability to lyse erythrocytes of sheep, rabbits, horses and humans. Five fractions, including intact cells, culture supernatant, culture filtrate, cell debris and lipid-enriched fractions, were prepared from the protozoan cells, and their hemolytic activities were assayed under various conditions. All the samples except culture supernatant had hemolytic activities, which were due to two different kinds of hemolysins. One hemolysin was protein-like and mainly found in cell-free fractions: culture supernatant and culture filtrate. It was heat-labile and inhibited by various cysteine-proteinase inhibitors. The other hemolysin was lipid-like and found in cell-associated fractions: intact cells, cell-debris and lipid-enriched fractions. It was heat-stable, organic solvent-tolerant and unaffected by various proteinase inhibitors and stimulators. These results suggested that T. tenax ATCC 30207 possessed two distinct hemolysins, protein and lipid.

Animals↗

Characterization of the cathepsin B-like proteinases of Trichomonas tenax ATCC 30207.

An oral parasite Trichomonas tenax ATCC 30207 synthesizes and secretes various proteinases. By gelatin-SDS-PAGE, we found five proteinases bands (30, 37, 46, 51 and 60 kDa) in cell lysate and four bands (37, 45, 52 and 60 kDa) in culture filtrate. The proteinases hydrolyzed acid soluble type I collagen as well as gelatin. The enzymes were suggested to possess typical characteristics of cysteine proteinases based on the patterns of inhibition and activation by various factors. Based on relative efficiencies of synthetic substrates, most of them were most likely cathepsin B-like enzymes.

Animals↗

Structure determination of [Arg8]vasopressin methylenedithioether in dimethylsulfoxide using NMR.

The structure of [Arg8]vasopressin methylenedithioether ([AVP]CH2) has been determined in dimethylsulfoxide-d6. Two-dimensional DQF-COSY and NOESY spectra were measured and used to derive angle and distance constraints for restrained molecular dynamics (MD) calculations. In the MD trajectory, two types of beta-turn structure were found in the region from Tyr2 to Asn5, suggesting an equilibrium between type-I and type-II' beta-turn structures. When Halpha chemical shifts were used as an additional constraint, the type-I turn was favoured. To validate this result, an independent energy minimization procedure was used, using differences between calculated and observed chemical shifts. The two approaches gave essentially identical results. It is therefore concluded that the type-I turn predominates in solution. Analysis of calculated chemical shift contributions suggests that the beta-turn structure found in AVP is well preserved in [AVP]CH2, although the pressin ring size is expanded.

Amino Acid Sequence↗

The application of the atomic force microscope to studies of medically important protozoan parasites.

Both living and fixed specimens of the medically-important parasitic protozoa, Trypanosoma cruzi, Toxoplasma gondii, Giardia lamblia, Entamoeba histolytica, and Acanthamoeba spp. were studied by atomic force microscopy (AFM). The preparation of fixed specimens was similar to methods used for either scanning or transmission electron microscopy. AFM scanning was performed using both contact and tapping modes. A classical fixation procedure utilizing glutaraldehyde followed by ethanol dehydration was not suitable for all parasite species. AFM images could not be obtained from fixed samples of T. cruzi, T. gondii or E. histolytica. However, excellent topographic images could be obtained from specimens of G. lamblia and Acanthamoeba under identical conditions. Critical point drying permitted AFM imaging of both trypomastigote and epimastigote stages of T. cruzi. Phase imaging of T. cruzi elucidated unique surface details at a level of resolution not visible using any other imaging modalities. AFM elasticity map imaging of T. cruzi-infected and T. gondii-infected cells demonstrated that both parasites were markedly firmer than the surrounding host cell cytoplasm. The parasitophorous vacuole surrounding replicating T. gondii tachyzoites was also visualized by elasticity map imaging. These data suggest that although much remains to be learned about preparing parasitic protozoa for AFM imaging, the technique has the potential of providing unique and important insights into these disease causing organisms.

Amoebida↗

The cell biological application of carbon nanotube probes for atomic force microscopy: comparative studies of malaria-infected erythrocytes.

We describe the first cell biological application of carbon nanotube (CN) probes for atomic force microscopy studies. Topographic and phase images were collected from Plasmodium falciparum malaria-infected erythrocytes using both TappingMode Etched Silicon Probes (TESP probe) and CN probes. We estimate that the lateral resolution of a CN probe-generated topographic image is at least four-fold higher than that of the TESP probe. Carbon nanotube probe-generated phase images of P. falciparum-induced knobs on the surface of erythrocytes also show a markedly higher lateral resolution than comparable TESP probe-generate phase images of the same area. We conclude that CN probes are useful for cell biological atomic force microscopy studies and should play an increasingly important role in the future of this evolving discipline.

Animals↗

[Guideline for the treatment of Hansen's disease in Japan].

Ad hoc committee of Japanese Leprosy Association recommends standard treatment protocol of leprosy in Japan, which is a modification of World Health Organization's multidrug therapy (WHO/MDT, 1997). For paucibacillary (PB) leprosy, 6 months treatment by rifampicin and dapsone (MDT/PB) is enough. However, for high bacterial load multibacillary (MB) leprosy, 12 months treatment seems insufficient. Thus, (A) For MB with bacterial index (BI) > or = 3 before treatment, 2 years treatment by rifampicin, dapsone and clofazimine (MDT/MB) is necessary. (A-1) When satisfactory decrease of BI (BI value decrease > or = 2 steps, or final BI < 3) is obtained after completion of 2 years MDT/MB, maintenance therapy by dapsone and clofazimine is recommended until BI negativity and loss of active lesions. (A-2) When BI decrease is not satisfactory (BI value decrease < 2 steps, or final BI > or = 3), MDT/MB should be continued until BI negativity and loss of active lesions. (B) For MB with BI < 3 or fresh MB (less than 6 months after the onset of the disease) with BI > or = 3, 1 year treatment by rifampicin, dapsone and clofazimine (MDT/MB) is necessary. (B-1) When BI become negative and active lesion is lost within one year, no maintenance therapy is necessary. (B-2) When BI is still positive or active lesion is remaining, additional therapy with MDT/MB for one more year is recommended. Brief summary of diagnosis, purpose of therapy, character of drugs, and prevention of deformity is also described.

Drug Therapy, Combination↗

Phase imaging by atomic force microscopy: analysis of living homoiothermic vertebrate cells.

Atomic force microscope-based phase imaging in air is capable of elucidating variations in material properties such as adhesion, friction, and viscoelasticity. However, the interpretation of phase images of specimens in a fluid environment requires clarification. In this report, we systematically analyzed atomic force microscope-derived phase images of mica, glass, and collagen under the same conditions as used for living cells at various tapping forces; the resulting data provide critical information for the interpretation of phase images of living cells. The peripheral regions of COS-1 cells consistently show a more negative phase shift than the glass substrate in phase images at set-point amplitude: free amplitude (Asp/A0) = 0.6-0.8. In addition, at all Asp/A0 values suitable for phase imaging, tapping frequency appears to be high enough to ensure that phase shifts are governed primarily by stiffness. Consequently, phase imaging is capable of high resolution studies of the cellular surface by detecting localized variations in stiffness. We demonstrate that phase imaging of a bifurcating fiber in COS-1 cell cytoplasm is readily capable of a lateral resolution of approximately 30 nm.

Aluminum Silicates↗

Kinetic analysis of the mitotic cycle of living vertebrate cells by atomic force microscopy.

The atomic force microscope (AFM) is becoming an important tool for qualitative and quantitative analyses of biological material. However, the difficulties involved in maintaining long-term, steady-state physiologic conditions and the problems associated with analyzing force curves generated from highly viscoelastic biological structures impede the use of the AFM for studies of kinetic processes in living vertebrate cells. In this report, we describe a simple method to track reproducibly kinetic changes in the localized stiffness of vertebrate cells. We tested our method on a study of vertebrate cells in mitosis and found a marked but transient decrease in stiffness occurs in the mitotic spindle region during anaphase. We propose that physical-chemical changes in the mitotic apparatus, most probably, changes in the state of polymerization of interzonal spindle fibers which also have been reported to undergo a marked reduction in birefringence during anaphase, are responsible for the observed decrease in stiffness. Our methodology affords a new approach to studying mitotic events and should be applicable to studies of a variety of viscoelastic properties of living cells.

Anaphase↗

An integrated approach to the study of living cells by atomic force microscopy.

We describe a technique for studying living cells with the atomic force microscope (AFM) in tapping mode using a thermostated, controlled-environment culture system. We also describe the integration of the AFM with bright field, epifluorescence and surface interference microscopy, achieving the highest level of integration for the AFM thus far described. We succeeded in the continuous, longterm imaging of relatively flat but very fragile cytoplasmic regions of COS cells at a lateral resolution of about 70 nm and a vertical resolution of about 3 nm. In addition, we demonstrate the applicability of our technology for continuous force volume imaging of cultured vertebrate cells. The hybrid instrument we describe can be used to collect simultaneously a diverse variety of physical, chemical and morphological data on living vertebrate cells. The integration of light microscopy with AFM and steady-state culture methods for vertebrate cells represents a new approach for studies in cell biology and physiology.

Animals↗

[The authentication of modern Japanese acupuncture].

The Meiji government that modernized Japan decided the direction which adopted western medicine positively and eliminated Oriental medicine, and carried out the policy to inhibit acupuncture (1874). However, there was no concrete system until 1885, when the Ministry of Home Affairs issued a report to entrust the permission and superintendance of acupuncture to each prefecture. On the other hand, modern education for the blind was begun in various parts of the country in 1878, and acupuncture was adopted as a vocational course. The Rakuzenkai Blind School began Anma and acupuncture in 1881, but in 1885 acupuncture was removed from the curriculum and put under the direct control of the Ministry of Education. The principal of the school, Ryokichi Yatabe, had doubts about this and sent a question to Tokyo Imperial University. The answer to it (1887) was to recognize acupuncture, and this recognition became an admission of the revival of acupuncture in the course of blind education and the basis of the thought in legislation for modern Japanese acupuncture. There is a high possibilty that the reply to Yatabe was influenced by the article of Sansaku Okumura (1864-1912), a blind man in Kanazawa City, which appeared in the 157th issue of "Iji Shimbun" (1885).

Acupuncture Therapy↗