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Recently, a new technique was developed for non-catalytically mixing microdroplets. In this method, an alternating-current (AC) electric field is used to promote the antigen–antibody reaction within the microdroplet. Previously, this technique has only been applied to histological examinations of flat structures, such as surgical specimens. In this study, we applied this technique for the first time to immunofluorescence staining of three-dimensional structures, specifically, mammalian eggs. We diluted an antibody against microtubules from 1:1,000 to 1:16,000, and compared the chromatic degree and extent of fading across dilutions. In addition, we varied the frequency of AC electric-field mixing from 5 Hz to 46 Hz and evaluated the effect on microtubule staining. Microtubules were more strongly stained after AC electric-field mixing for only 5 minutes, even when the concentration of primary antibody was 10 times lower than in conventional methods. AC electric-field mixing also alleviated microtubule fading. At all frequencies tested, AC electric-field mixing resulted in stronger microtubule staining than in controls. There was no clear difference in a microtubule staining between frequencies. These results suggest that the novel method could reduce antibody consumption and shorten immunofluorescence staining time.

A protective glycolipid antigen (PAg) was extracted from Leptospira interrogans serovar canicola with chloroform/methanol/water (1 : 2: 0.8, by vol.) and partially purified by silica gel column chromatography. The PAg elicited a protective response in hamsters and in cyclophosphamide-treated mice subsequently challenged with homologous Lepfospira. The PAg band was detected as a single smear-like band, corresponding to a protein of 23-30 kDa, by silver-staining in SDS-PAGE. In immunoblots, this band reacted with a monoclonal antibody, A5, which agglutinated serovar cunicola and recognized a serovar-specific antigen. Furthermore, the PAg did not migrate on silica gel TLC, but was detected at the origin as a ninhydrin-and naphthol-positive spot. This suggests that PAg is a hydrophilic molecule with a carbohydrate chain that contains amino groups, possibly as amino sugars.

AimTo describe the registry design of the Osaka Emergency Information Research Intelligent Operation Network system (ORION) and its profile of hospital information, patient and emergency medical service characteristics, and in‐hospital outcomes among all patients transported to critical care centers and emergency hospitals in Osaka Prefecture, Japan.MethodsThe Osaka Prefecture Government has developed and introduced an information system for emergency patients (the ORION system) that uses a smartphone application (app) for hospital selection by on‐scene emergency medical service personnel and has been accumulating all ambulance records. Since January 2015, medical institutions have obtained information on the diagnosis and outcome of patients transported to medical institutions, and the ORION system merged these data with ambulance records including smartphone app data.ResultsFrom January 2015 to December 2016, 753,301 eligible patients were registered. The mean age was 58.7 years, and 51.5% of patients were male. After hospital arrival, 39.7% were hospitalized, 58.2% were discharged from hospital, 1.1% changed hospital, and 1.0% died. The most common diagnoses were injury, poisoning, and certain other consequences of external causes. Among the hospitalized patients, 29.2% were continuously hospitalized, 59.0% discharged, 5.2% changed hospital, and 5.8% were dead at 21 days after hospitalization. The most common confirmed diagnosis was diseases of the circulatory system.ConclusionUsing the ORION system developed and operated by Osaka Prefecture since January 2015, we described the epidemiological data of all emergency patients transported to emergency hospitals. Analysis using the ORION database in the future could lead to improvements in the emergency transport system and patient outcomes.

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