Culture of rabbit middle ear epithelial cells. A method for primary culture and subculture with identification, characterization and growth specification

During the last decade middle ear epithelium has been cultured from various species. Until now, subcultivation has been achieved only with the use of a feeder-cell layer or conditioned medium. These factors are possible confounders in the in vitro model. On the other hand, subcultivation is necessary for exact quantitative studies. We present a reproducible culture method allowing subcultivation without feeder-cells or conditioned medium. The main features in our method are a low-serum, hormone-supplemented medium, an incubation temperature of 34 degrees C, fixation of explants, gentle trypsinization and replating with high cell density. Cells were identified by immunohistochemistry through a battery of monclonal antibodies. The percentage of epithelial cells in the subculture was 99.2%. To our knowledge, this is the first report describing subcultivation of middle ear epithelial cells exclusively in a completely controlled environment. These are optimal circumstances for future investigation and quantification of various factors influencing proliferation and differentiation of middle ear epithelium.     

New technologies for amperometric biosensors

Amperomeric-based detectors have successfully been used as personal monitors for blood glucose levels. However, there is a desire to increase the number of compounds measured in a small blood sample, the speed of detection and enhance the reliability of the measurement. Furthermore, with the increasing use of microdialysis as a clinical sampling method in metabolic medicine, paediatric medicine and neurointensive care, there is a need for rapid on-line detection of analytes such as lactate, glucose and glutamate in low microlitre volume samples. Two approaches to these problems are described. The first uses enzymes immobilized in a packed bed with electrochemical detection of a ferrocene mediator as a flow-injection assay for use with microdialysis. Results from microdialysis of the brain of freely moving rats are described. In the second approach, thin-film techniques are used to fabricate arrays of microdisk and micro line electrodes. The properties of these arrays in free solution and in a flow cell are presented together with an example using multiple arrays to identify an analyte by oxidation potential. Finally, different enzymes are entrapped onto the surface of two arrays by electrochemical polymerization of o-phenylenediamine. The resulting device detects glucose and lactate in real-time.     

The preparation and hydrolysis of silicon isopropoxides

The preparation of tetraisopropoxysilane by the liquid phase reaction between tetrachlorosilane and isopropanol is described, together with the preparation of ‘isopropyl silicate’ by treating tetrachlorosilane or crude tetraisopropoxysilane with a mixture of isopropanol and water in which the molar ratio SiCl₄: H₂O is less than 1:1. This ‘isopropyl silicate’ is a mixture of tetraisopropoxysilane and isopropoxypolysiloxanes. The formation of isopropoxypolysiloxanes from pure tetraisopropoxysilane saturated with hydrogen chloride is also described. The hydrolysis of silicon isopropoxides is discussed. Under suitable conditions, hydrolysates which may be used to bind powdered refractories can be obtained.     

Selection for agonistic begavior in wild female Mus musculus

Artificial selection was used to establish different levels of agonistic behavior in laboratory-reared wild female Mus musculus. A within-family selection design with replicated high and low lines and two nonselected control lines was employed. Females only were tested at 8 weeks of age on 2 consecutive days. Testing consisted of placing a C57BL/6 female mouse in the home cage of the isolation-reared wild mouse for 7 min or until an attack occurred. The wild females were rated on a 5-point scale for agonistic behavior, and the sum of the scores over the 2 test days was the criterion for selection. The six lines, each containing ten breeding pairs, were selected for four generations. By the fourth selected generation, the responses of the high and low lines had diverged in the expected directions. One-way analyses of variance indicated reliable differences among high, low, and control lines in the second, third, and fourth generations. It was concluded that female agonistic behavior is influenced by genotype and that the level of this behavior can be manipulated by means of artificial selection.     

Reaction between chlorophosphazenes and some alkyl phosphates: Formation of temperature-stable thermosetting materials

The reaction between chlorophosphazenes and triethyl, tri-n-butyl and acid ethyl phosphates has been studied as a means of preparing temperature-stable thermosetting resinous materials. Ethyl or n-butyl chlorides were eliminated (together with hydrogen chloride in the case of acid ethyl phosphate) and thermosetting resinous products were obtained. The products are film forming when stoved in air. Asbestos laminates, which retain up to 64% of their strength when heated to 500 °C for 50 h in air, have been made with the thermosetting resinous products obtained from the reaction between a chlorophosphazene and triethyl or tri-n-butyl phosphates.