Isolation of Acanthamoeba with Sabouroud's glucose agar plate

Isolation of Acanthamoeba or finding out the cysts from corneal leasions is necessary to confirm the diagnosis of Acanthamoeba keratitis. However, we cannot always find out the cyst from corneal scrapings of the lesions. In these cases the isolation of Acanthamoeba is very important to confirm the diagnosis. Non-nutrient agar plates are generally used to isolate Acanthamoeba, but it is difficult to get them easily at many hospitals. To find out another agar that can be used to isolate Acanthamoeba and got easily, Sabouroud's glucose agar plates were compared with non-nutrient agar plates for the isolation of Acanthamoeba. Both agar plates seeded with Escherichia coli showed a nice growth of trophozoites first and cysts later. A clinical material, stock solution of contact lens used by a suspected patient of Acanthamoeba keratitis, was put on Sabouroud's glucose agar plate seeded with Escherichia coli. After three days of incubation at 25°C Acanthamoeba was found on the plate. Sabouroud's glucose agar plate was a very useful agar plate for the isolation of Acanthamoeba

Protection of mice from LPS-induced shock by CD14 antisense oligonucleotide.

CD14 is a pattern recognition receptor on myeloid cells and plays a pivotal role in an innate immune system that is responsible for Gram-negative and Gram-positive bacteria infection. Lipopolysaccharide (LPS), a cell wall component of Gram-negative bacteria, can induce production of a large quantity of proinflammatory cytokines into the circulation mediated by CD14-mediated macrophages and monocytes. These cytokines eventually cause septic shock. Several in vitro and in vivo studies have shown that suppression of a CD14 function by a CD14 antibody led to an inhibition of the production of proinflammatory cytokines such as TNF-alpha, IL-1 beta, and IL-8. In the present study, we found that CD14 antisense oligonucleotide (ODN) can prevent lethal LPS shock in D-galactosamine-sensitized mice. This ODN inhibited CD14 expression in a mouse macrophage cell line, RAW264.7, and suppressed production of TNF-alpha in LPS-stimulated RAW264.7 cells. Furthermore, we designed a consensus antisense ODN that could hybridize human and mouse CD14 RNA, and we evaluated its efficacy. The consensus antisense ODN rescued mice primed with Mycobacterium bovis bacillus Calmette-Guerin (BCG) from the LPS-induced lethal shock. In this model, the CD14 antisense ODN down-regulated LPS-elicited CD14 expression in the liver, resulting in a decrease in LPS-induced TNF-alpha production. These findings suggest that the CD14 antisense ODN is distributed in the liver and efficiently suppresses LPS-induced TNF-alpha production by reducing CD14 expression on Kupffer cells. This CD14 antisense ODN may be useful for the development of a therapeutic agent against sepsis and septic shock.</p

Electron microscopic studies of Acanthamoeba : 2 Morphological changes of cultured Acanthamoeba due to anti-amoebic agents

Morphological changes of Acanthamoeba in Peptone-Yeast-Glucose medium containing anti-amoebic agents were studied with transmission electron microscope. In fradiomycin-treated amoeba, decrease of acanthopodia, vacuolar formation in the ectoplasm and cell collapse were observed. In polyhexamethylene biguanide (PHMB)-treated amoeba, rupture of plasma membrane and degeneration of mitochondria were observed at the early stage. These results indicate that PHMB is more effective than fradiomycin for the treatment of Acanthaoeba keratitis

Electroencephalographic changes during piano playing and related mental tasks.

Theta activity in EEG was found to be augmented in the frontal midline area in 5 young women while they played classical piano pieces and during related mental tasks. This activity was considered to be a frontal midline theta rhythm with the maximal amplitude mostly in Fz and of the frequency ranging from 5 to 7.5 Hz. This theta activity was observed to increase depending on the degree of the subjects' concentration on piano playing or related tasks. In bilateral parietal derivations, increases in the power value of alpha activity were observed in some subjects while they were listening to music, suggesting that alpha activity was involved in appreciation of music.</p

Papillo-Choledochectomy in the Operative Management of Mucosal Neoplasms of the Periampullary Region

Two patients with mucosal cancer of the periampullary region were treated with papillocholedochectomy, which entails removal of the papilla of Vater and the whole length of the common bile duct. The neoplasm is dissected out through the plane between the duodenal circular and longitudinal muscles, deep to the sphincter of Oddi and the fibromuscular layer of the bile duct. Pathological examination showed that cancer was confined to the mucosal layer without stromal invasion, and that the operation achieved radical cure. For mucosal cancer, papillo-choledochectomy is an alternative to pancreatoduodenectomy, provided that repeated frozen-section studies confirm the completeness of excision

Relevance of Some Damage Factors to Structures Damage in the 1995 Kobe Earthquake

The 1995 Kobe Earthquake (Hyogoken-nanbu Earthquake) caused a severe damage to various kinds of structures. As damage factors of these structures, the characteristics of seismic motion, distance from the earthquake source fault, ground conditions, liquefaction, and strength of structures can be considered. In this paper, paying attention to the distance from the earthquake source fault and ground conditions among them, the relevance to structures damage in wooden houses (on-ground structure) and water supply pipelines (under-ground structure) in Nishinomiya-City area was examined. As the results, the relationship between wooden houses damage and the distance from the fault can be approximately represented as a unique exponential function. In liquefied areas, however, the rate of completely collapsed wooden houses decreases 5 to 20% from the average value. This might be because the damping of earthquake motion brought the decrease of the damage rate. While, the relationship between water supply pipelines damage and the distance from the fault completely differs from the above-mentioned for wooden houses. A characteristic value Tg estimated from the distribution of N value at each location can be used for the ground classification in earthquake-proof design. The damage rate of water supply pipelines increases as increasing Tg , while that of wooden houses decreases as increasing Tg

AKARI infrared imaging of reflection nebulae IC4954 and IC4955

We present the observations of the reflection nebulae IC4954 and IC4955 region with the Infrared Camera (IRC) and the Far-Infrared Surveyor (FIS) on board the infrared astronomical satellite AKARI during its performance verification phase. We obtained 7 band images from 7 to 160um with higher spatial resolution and higher sensitivities than previous observations. The mid-infrared color of the S9W (9um) and L18W (18um) bands shows a systematic variation around the exciting sources. The spatial variation in the mid-infrared color suggests that the star-formation in IC4954/4955 is progressing from south-west to north-east. The FIS data also clearly resolve two nebulae for the first time in the far-infrared. The FIS 4-band data from 65um to 160um allow us to correctly estimate the total infrared luminosity from the region, which is about one sixth of the energy emitted from the existing stellar sources. Five candidates for young stellar objects have been detected as point sources for the first time in the 11um image. They are located in the red S9W to L18W color regions, suggesting that current star-formation has been triggered by previous star-formation activities. A wide area map of the size of about 1 x 1 (deg^2) around the IC4954/4955 region was created from the AKARI mid-infrared all-sky survey data. Together with the HI 21cm data, it suggests a large hollow structure of a degree scale, on whose edge the IC4954/4955 region has been created, indicating star formation over three generations in largely different spatial scales.Comment: 23 pages, 7 figures, accepted for publication in PASJ AKARI special issu

Substance use and sexual behaviours of Japanese men who have sex with men: A nationwide internet survey conducted in Japan

BACKGROUND: Japanese men who have sex with men (MSM), especially those living in large metropolitan areas such as Tokyo and Osaka, are facing a growing HIV/AIDS epidemic. Although the Internet is used as a new venue for meeting sex partners, it can also serve as a useful research tool for investigating the risk behaviours of Japanese MSM. This Internet survey explored the extent of substance use and its association with sexual risk behaviours among Japanese MSM. METHODS: Between 28 February 2003 and 16 May 2003 MSM were recruited through 57 Japanese gay-oriented Web sites, gay magazines, and Internet mailing lists. Participants completed a structured questionnaire anonymously through the Internet. RESULTS: In total, 2,062 Japanese MSM completed the questionnaire. The average age of participants was 29.0 years and 70.5% identified as gay, 20.8% as bisexual, and 8.7% as other. Overall, 34.5% reported never using a substance, 45% reported ever using one type of substance (lifetime reported single substance users), and 19.6% had used more than 1 type of substance (lifetime reported multiple substance users) in their lifetimes. The substances most commonly used were amyl nitrite (63.2%), 5-methoxy-N, N-diisopropyltryptamine (5MEO-DIPT) (9.3%), and marijuana (5.7%). In the multivariate analysis, unprotected anal intercourse, having had 6 or more sexual partners, visiting a sex club/gay venue in the previous 6 months, a lower education level, and being 30 to 39 years of age were associated with both lifetime single and lifetime multiple substance use. Lifetime reported multiple substance use was also correlated with having a casual sex partner, having symptoms of depression, being diagnosed as HIV-positive, and greater HIV/AIDS-related knowledge. CONCLUSION: This is the first Internet-based research focused on the sexual and substance use behaviours of MSM in Asia. Our findings suggest a compelling need for prevention interventions to reduce HIV risk-related substance use behaviours among Japanese MSM. The results also suggest that the Internet is potentially a useful tool for collecting behavioural data and promoting prevention interventions among this population

Quantitative Analysis of Serum Procollagen Type I C-Terminal Propeptide by Immunoassay on Microchip

BACKGROUND: Sandwich enzyme-linked immunosorbent assay (ELISA) is one of the most frequently employed assays for clinical diagnosis, since this enables the investigator to identify specific protein biomarkers. However, the conventional assay using a 96-well microtitration plate is time- and sample-consuming, and therefore is not suitable for rapid diagnosis. To overcome these drawbacks, we performed a sandwich ELISA on a microchip. METHODS AND FINDINGS: The microchip was made of cyclic olefin copolymer with straight microchannels that were 300 µm wide and 100 µm deep. For the construction of a sandwich ELISA for procollagen type I C-peptide (PICP), a biomarker for bone formation, we used a piezoelectric inkjet printing system for the deposition and fixation of the 1st anti-PICP antibody on the surface of the microchannel. After the infusion of the mixture of 2.0 µl of peroxidase-labeled 2nd anti-PICP antibody and 0.4 µl of sample to the microchannel and a 30-min incubation, the substrate for peroxidase was infused into the microchannel; and the luminescence intensity of each spot of 1st antibody was measured by CCD camera. A linear relationship was observed between PICP concentration and luminescence intensity over the range of 0 to 600 ng/ml (r(2) = 0.991), and the detection limit was 4.7 ng/ml. Blood PICP concentrations of 6 subjects estimated from microchip were compared with results obtained by the conventional method. Good correlation was observed between methods according to simple linear regression analysis (R(2) = 0.9914). The within-day and between-days reproducibilities were 3.2-7.4 and 4.4-6.8%, respectively. This assay reduced the time for the antigen-antibody reaction to 1/6, and the consumption of samples and reagents to 1/50 compared with the conventional method. CONCLUSION: This assay enabled us to determine serum PICP with accuracy, high sensitivity, time saving ability, and low consumption of sample and reagents, and thus will be applicable to clinic diagnosis