Accomplishments of the Trustees and laboratory staff of the Biological Stain Commission, 2002–2013

During the 12 years from 2002 to 2013, the Trustees and laboratory personnel of the Biological Stain Commission (BSC) can claim many accomplishments. These accomplishments are itemized under 11 categories: continuous publication of the official journal, Biotechnic & Histochemistry; production of four special issues of Biotechnic & Histochemistry devoted to specific dyes or stains; standardization of staining and dye purity; mechanisms of staining and prediction of dye behavior; publication of books or book chapters; effects of fixation and processing on staining; cancer research; immunohistochemistry; BSC Laboratory activities; miscellaneous publications; and administrative accomplishments.     

The growth of individual seminal vesicle epithelial cells and their proliferation

Histologically seminal vesicle epithelium (SVE) of the intact adult guinea pig is a discrete and segregated monolayer of highly specialized tall columnar cells. The epithelial layer is so sharply demarcated from its attached stroma (primarily smooth muscle), that blunt dissection alone is sufficient to separate epithelium from muscle. After castration the epithelial cells decrease in both size and number so that by the fifth day, the surviving cells are greatly involuted structurally and comprise only about 12% of the original numerical population normally present in one seminal vesicle. Injected testosterone leads to restructuring of individual cells followed by cell replenishment. The major goal of this effort was to elaborate upon the processes of individual cell growth and cell replenishment during restoration of the tissue to normal cell size and number. The two separate processes were studied using light and electron microscopy, [3H]thymidine incorporation, and Northern blots with labeled histone gene probes. By approximately 48 hr of hormone repletion, parenchymal cell size had returned to normal as the result of a dramatic anabolic process of individual cell growth while cell number remained unchanged. During the subsequent 48-hr period of hormone repletion, the cell population was restored to normal as cell replenishment became the predominant process. Microscopic analysis at intervals throughout the 96-hr period failed to disclose any mitotic events to account for cell replenishment even when Colcemid had been administered. Nor could the increase in cell numbers be correlated with a great increase in [3H]thymidine incorporation or in histone mRNA synthesis. Thus, we could provide no evidence that mitotic division of the parenchymal cells themselves is responsible for cell replenishment. During the 24- to 48-hr interval of hormone repletion, electron microscopic examination disclosed the presence of small epithelial cells lying in a basal position. Some of these cells were seen to insert themselves between the basal regions of parenchymal cells and to expand from the basement membrane into the parenchyma. Possible origins of the cells which replenish the tissue are discussed.     

Concentration of liver and kidney fructose-1,6-diphosphatase determined by specific radioimmunoassay

A radioimmunoassay for liver fructose-1,6-diphosphatase (D-fructose-1,6-bisphosphate 1-phosphohydrlase, EC 3.1.3.11) has been developed based on maintenance of its tetrameric structure and immunologic integrity after iodination by the Bolton-Hunter technique. The assay detected as little as 2 ng of standard enzyme. Nonspecific interference by tissue components did not occur. Enzyme concentration (mumol/1000 g tissue wet weight) was measured in tissue extracts of 49 rabbits subjected to a variety of conditions. In animals fed a 'balanced' diet containing 50--60% carbohydrate (by weight), the concentration in liver was 3.4 microM +/- 0.3. After fasts of 48, 72, or 96 h, the concentration in liver increased approximately 1.4-fold. A high-fat diet did not alter the concentration significantly but a high-protein diet caused an increase of 2.1-fold to 7.2 microM +/- 1.4. The greatest concentrations, 8.7 microM +/- 1.9, were observed in the livers of severely diabetic rabbits. The increase paralleled the increasing severity of diabetes and provides one explanation for the augmented gluconeogenesis which occurs in the diabetic state. Changes were less marked in kidney. The greatest apparent incrase, from 2.6 microM +/- 1.1 in the normal fed rabbit to 4.7 microM +/- 2.8, occurred in the severely diabetic animal. However, variation was sufficiently great in kidney to render apparent increases during fasting, protein feefing and diabetes statistically insignificant. For the most part changes in assayable activity followed changes in enzyme concentration except in the rabbits maintained on high-protein diets. In these, liver enzyme concentration increased by 2.4-fold whereas activity increased by only 1.3-fold, and the kidney enzyme concentration increased 1.3-fold whereas activity decreased by 20%.     

Progress in treatment of ANCA-associated vasculitis

Autoantibodies to neutrophil cytoplasmic antigen-associated vasculitis (AAV) is characterised by inflammation of blood vessels. The introduction of immunosuppressive therapy with glucocorticoids and cyclophosphamide transformed AAV from a fatal condition to a largely treatable condition. Over the past 30 years, considerable progress has been made refining immunosuppressive regimens with a focus on minimising toxicity. There is, however, a high unmet need in the treatment of AAV. A proportion of patients are refractory to current therapies; 50% experience a relapse within 5 years and treatment toxicity contributes to mortality and chronic disability. As knowledge of the pathogenesis of vasculitis grows, it is mirrored by the availability of biological agents, which herald a revolution in the treatment of vasculitis. Lymphocyte-targeted and cytokine-targeted agents have been evaluated for the treatment of AAV and are entering the routine therapeutic arena with the potential to improve patient outcomes. As rare diseases, treatment advances in vasculitis depend on international collaborative research networks both to establish an evidence base for newer agents and to develop recommendations for patient management.     

31P nuclear magnetic resonance spectroscopy study on kidney preservation. Effect of verapamil

31P NMR spectroscopy has been used to evaluate the usefulness of verapamil, a calcium channel blocker, in preventing ischemic renal damage. Phosphorylated metabolites have been investigated before, during and after 48 hrs of hypothermic storage. The rapidity in adenosine triphosphate resynthesis and the phosphomonoesters and phosphodiesters levels after reperfusion at the end of the storage period (48 hrs), were significantly higher in verapamil-treated kidneys. Phosphomonoesters to inorganic phosphate ratio, during the storage period, is even higher. These findings suggest that verapamil may protect against ischemic renal damage and so it can be useful for renal preservation. Furthermore, it has been shown that 31P NMR spectroscopy puts into evidence the biochemical recovery and allows the assessment of the viability of organs.