Ethyl pyruvate diminishes the endotoxin-induced inflammatory response of bovine mammary endothelial cells.

The endotoxin-induced inflammatory response during coliform mastitis is difficult to control with the currently available therapeutics. Endothelial cells are among the first cell type to be engaged in the inflammatory response and can modulate the severity of inflammation by producing proinflammatory mediators upon endotoxin exposure. Ethyl pyruvate, an ethyl ester of pyruvic acid, can ameliorate endotoxin-induced inflammatory responses by inhibiting the production of proinflammatory mediators in several in vitro and in vivo endotoxemia models. The objective of this study was to determine the effect of ethyl pyruvate on the production of vascular proinflammatory mediators that are associated with the pathogenesis of coliform mastitis. The ability of ethyl pyruvate to reduce the expression of proinflammatory mediators was evaluated in cultured bovine mammary endothelial cells (BMEC) stimulated with endotoxin. Treatment of endotoxin-stimulated BMEC with ethyl pyruvate significantly reduced gene expression of IL-6, IL-8, and intercellular adhesion molecule 1 as well as expression of eicosanoid-producing enzymes, including cyclooxygenase 2 and 15-lipoxygenase 1. This is the first time that the effect of ethyl pyruvate was evaluated in an in vitro BMEC model of coliform mastitis. The ability of ethyl pyruvate to effectively inhibit gene and protein expression of potent vascular proinflammatory mediators in vitro warrants further investigations to assess in vivo efficacy. Ethyl pyruvate is safe for human consumption, and it may be an attractive candidate as a therapeutic in ameliorating the severe pathogenesis associated with coliform mastitis.

A portable multi-flash kinetic fluorimeter for measurement of donor and acceptor reactions of Photosystem 2 in leaves of intact plants under field conditions.

A newly-developed field-portable multi-flash kinetic fluorimeter for measuring the kinetics of the microsecond to millisecond reactions of the oxidizing and reducing sides of photosystem 2 in leaves of intact plants is described and demonstrated. The instrumental technique is a refinement of that employed in the 'double-flash' kinetic fluorimeter (Joliot 1974 Biochim Biophys Acta 357: 439-448) where a low-intensity short-duration light pulse is used to measure the fluorescence yield changes following saturating single-turnover light pulses. The present instrument uses a rapid series of short-duration (2 µs) pulses to resolve a complete microsecond to millisecond time-scale kinetic trace of fluorescence yield changes after each actinic flash. Differential optics, using a matrix of optical fibers, allow very high sensitivity (noise levels about 0.05% Fmax) thus eliminating the need for signal averaging, and greatly reducing the intensity of light required to make a measurement. Consequently, the measuring pulses have much less actinic effect and an entire multi-point trace (seven points) excites less than 1% of the reaction centers in a leaf. In addition, bu combining the actinic and measuring pulse light in the optical fiber network, the tail of the actinic flash can be compensated for, allowing measurements of events as rapidly as 20 µs after the actinic flash. This resolution makes practical the routine measurement of the microsecond turnover kinetics of the oxygen evolving complex in leaves of intact plants in the field. The instrument is demonstrated by observing flash number dependency and inhibitor sensitivity of the induction and decay kinetics of flash-induced fluorescence transients in leaves of intact plants. From these traces the period-two oscillations associated with the turnover of the two-electron gate and the period-four oscillations associated with the turnover of the oxygen evolving complex can be observed. Applications of the instrument to extending our knowledge of chloroplast function to the whole plant, the effects on plants of environmental stress, herbicides, etc, and possible applications to screening of mutants are discussed.

Extrasplanchnic effect of sulfobromophthalein (BSP) on plasma estrogen levels in the dog.

The effect of the drug sulfobromophthalein (BSP) on plasma estrogens in the dog were studied during intravenous infusions of 3H-labeled estrogens. During [3H]estrone infusion, BSP administration caused a marked increase in arterial plasma levels of the radioactive conjugated estrogens, estrone glucosiduronate, estradiol-17 beta glucosiduronate(s), and estrone sulfate. Levels of the unconjugated estrogens, estrone and estradiol-17 beta, were substantially unaltered. Possible mechanisms were investigated. Splanchnic extraction of the conjugates did not change significantly during BSP administration, and renal excretion rose promptly in proportion to the plasma levels, thus virtually excluding decreased biliary or renal excretion. There was no net discharge of estrogen glucosiduronate radioactivity from adipose tissue or muscle following BSP. During [3H]estrone glucosiduronate infusion, BSP again caused an increase in plasma estrone glucosiduronate, thus excluding increased formation (of this conjugate, at least). BSP caused decreased extraction of estrone glucosiduronate by the hindlimb, indicating that decreased metabolism was the probable cause of the elevated plasma levels. BSP also caused decreased formation of unconjugated estrogens by the lungs, indicating that the decreased metabolism includes decreased hydrolysis of estrogen glucosiduronates.