Surface Visualisation of Tissue Interfaces by Scanning Electron Microscopy. Methods for Exposure of the Basal Lamina and Associated Structures in Human Amnion

Tissue interfaces such as basal lamina have been traditionally investigated in transmission electron microscopy by sections cut vertical to the lamina, presenting information restricted to a single ultrathin plane. In order to overcome this limitation, a methodology for surface visualisation of the underside cell membranes of the amniotic epithelium, the upper and lower basal lamina surfaces, and their relationship to the stromal collagen has been devised. This involves alkaline, detergent or enzymatic loosening and/or removal of the epithelial monolayer prior to fixation, followed by dry fracture after critical point drying. In this way we have visualised large areas of all interfaces and the inter-relationships between these elements during the process of stromal collagen production by the amniotic epithelial cells

Aspirated capacitor measurements of air conductivity and ion mobility spectra

Measurements of ions in atmospheric air are used to investigate atmospheric electricity and particulate pollution. Commonly studied ion parameters are (1) air conductivity, related to the total ion number concentration, and (2) the ion mobility spectrum, which varies with atmospheric composition. The physical principles of air ion instrumentation are long-established. A recent development is the computerised aspirated capacitor, which measures ions from (a) the current of charged particles at a sensing electrode, and (b) the rate of charge exchange with an electrode at a known initial potential, relaxing to a lower potential. As the voltage decays, only ions of higher and higher mobility are collected by the central electrode and contribute to the further decay of the voltage. This enables extension of the classical theory to calculate ion mobility spectra by inverting voltage decay time series. In indoor air, ion mobility spectra determined from both the novel voltage decay inversion, and an established voltage switching technique, were compared and shown to be of similar shape. Air conductivities calculated by integration were: 5.3 +- 2.5 fS/m and 2.7 +- 1.1 fS/m respectively, with conductivity determined to be 3 fS/m by direct measurement at a constant voltage. Applications of the new Relaxation Potential Inversion Method (RPIM) include air ion mobility spectrum retrieval from historical data, and computation of ion mobility spectra in planetary atmospheres.Comment: To be published in Review of Scientific Instrument

Poison sedge can kill stock

POISON SEDGE was first suspected of being toxic to livestock in Western Australia nearly 80 years ago. Sudden deaths of sheep grazing areas on which poison sedge grew have been reported from many regions from Geraldton to Scott River. This article reports a case of poisoning in the field, and the experimental reproduction of poison sedge toxicity in pen-fed sheep

Automated longwall guidance and control vertical control subsystem, volume 2

Software documentation and a computer program listing for a horizon control of a longwall shearer are presented

Characterisation of Osteopontin in an In Vitro Model of Embryo Implantation

At the onset of pregnancy, embryo implantation is initiated by interactions between the endometrial epithelium and the outer trophectoderm cells of the blastocyst. Osteopontin (OPN) is expressed in the endometrium and is implicated in attachment and signalling roles at the embryo–epithelium interface. We have characterised OPN in the human endometrial epithelial Ishikawa cell line using three different monoclonal antibodies, revealing at least nine distinct molecular weight forms and a novel secretory pathway localisation in the apical domain induced by cell organisation into a confluent epithelial layer. Mouse blastocysts co-cultured with Ishikawa cell layers served to model embryo apposition, attachment and initial invasion at implantation. Exogenous OPN attenuated initial, weak embryo attachment to Ishikawa cells but did not affect the attainment of stable attachment. Notably, exogenous OPN inhibited embryonic invasion of the underlying cell layer, and this corresponded with altered expression of transcription factors associated with differentiation from trophectoderm (Gata2) to invasive trophoblast giant cells (Hand1). These data demonstrate the complexity of endometrial OPN forms and suggest that OPN regulates embryonic invasion at implantation by signalling to the trophectoder

Osmotic stress induces JNK-dependent embryo invasion in a model of implantation

In vitro culture during assisted reproduction technologies (ARTs) exposes pre-implantation embryos to environmental stressors, such as non-physiological nutritional, oxidative and osmotic conditions. The effects on subsequent implantation are not well understood but could contribute to poor ART efficiency and outcomes. We have used exposure to hyperosmolarity to investigate the effects of stress on the ability of embryos to interact with endometrial cells in an in vitro model. Culturing mouse blastocysts for 2 h in medium with osmolarity raised by 400 mosmol induced blastocoel collapse and re-expansion, but did not affect subsequent attachment to, or invasion of, the endometrial epithelial Ishikawa cell line. Inhibition of stress-responsive c-Jun N-terminal kinase (JNK) activity with SP600125 did not affect the intercellular interactions between these embryos and the epithelial cells. Four successive cycles of hyperosmotic stress at E5.5 had no effect on attachment, but promoted embryonic breaching of the epithelial cell layer by trophoblast giant cells in a JNK-dependent manner. These findings suggest that acute stress at the blastocyst stage may promote trophoblast breaching of the endometrial epithelium at implantation and implicates stress signalling through JNK in the process of trophectoderm differentiation into the invasive trophoblast necessary for the establishment of pregnancy. The data may lead to increased understanding of factors governing ART success rates and safety