Effects of ovariectomy of the dogfish Scyliorhinus canicula L. on circulating levels of androgen and oestradiol and on pituitary gonadotrophin content

Levels of androgens and oestradiol in dogfish, Scyliorhinus canicula , plasma, measured by radioimmunoassay, decrease markedly after ovariectomy (OVX). The magnitude and latency of these responses varies according to length of captivity preceding the operation and the time of year at which ovariectomy is performed. Androgens are cleared more rapidly than oestradiol from plasma following OVX, and levels of both steroids are reduced after laparotomy (LAP) in November but not in March. An extract of ventral lobes of the dogfish pituitary induces a significant increase in the circulating androgen levels in LAP fish but not in OVX fish, indicating that the ovary is the major site of androgen production in response to stimulation by dogfish gonadotrophin. Intramuscular injections of oestradiol benzoate (2.5 mg per fish) into LAP fish produce a significant reduction in the gonadotrophin content of the pituitary ventral lobe, measured in the quail testicular cell bioassay, when compared to LAP and unoperated controls. However, no significant changes in the gonadotrophin content of the ventral lobe were seen five days after OVX in June.     

Spatial co‐localisation of extreme weather events: a clear and present danger

Extreme weather events have become a dominant feature of the narrative surrounding changes in global climate with large impacts on ecosystem stability, functioning and resilience; however, understanding of their risk of co‐occurrence at the regional scale is lacking. Based on the UK Met Office’s long‐term temperature and rainfall records, we present the first evidence demonstrating significant increases in the magnitude, direction of change and spatial co‐localisation of extreme weather events since 1961. Combining this new understanding with land‐use data sets allowed us to assess the likely consequences on future agricultural production and conservation priority areas. All land‐uses are impacted by the increasing risk of at least one extreme event and conservation areas were identified as the hotspots of risk for the co‐occurrence of multiple event types. Our findings provide a basis to regionally guide land‐use optimisation, land management practices and regulatory actions preserving ecosystem services against multiple climate threats.     

Isolation and characterisation of human pulmonary microvascular endothelial cells from patients with severe emphysema

Background

Loss of the pulmonary microvasculature in the pathogenesis of emphysema has been put forward as a credible alternative to the classical inflammatory cell driven proteolysis hypothesis. Mechanistic studies in this area have to date employed animal models, immortalised cell lines, primary endothelial cells isolated from large pulmonary arteries and non-pulmonary tissues and normal human pulmonary microvascular endothelial cells. Although these studies have increased our understanding of endothelial cell function, their relevance to mechanisms in emphysema is questionable. Here we report a successful technique to isolate and characterise primary cultures of pulmonary microvascular endothelial cells from individuals with severe emphysema.

Methods

A lobe of emphysematous lung tissue removed at the time of lung transplantation surgery was obtained from 14 patients with severe end-stage disease. The pleura, large airways and large blood vessels were excised and contaminating macrophages and neutrophils flushed from the peripheral lung tissue before digestion with collagenase. Endothelial cells were purified from the cell mixture via selection with CD31 and UEA-1 magnetic beads and characterised by confocal microscopy and flow cytometry.

Results

Successful isolation was achieved from 10 (71%) of 14 emphysematous lungs. Endothelial cells exhibited a classical cobblestone morphology with high expression of endothelial cell markers (CD31) and low expression of mesenchymal markers (CD90, αSMA and fibronectin). E-selectin (CD62E) was inducible in a proportion of the endothelial cells following stimulation with TNFα, confirming that these cells were of microvascular origin.

Conclusions

Emphysematous lungs removed at the time of transplantation can yield large numbers of pulmonary microvasculature endothelial cells of high purity. These cells provide a valuable research tool to investigate cellular mechanisms in the pulmonary microvasculature relevant to the pathogenesis of emphysema.