W. W. "Mo" Cleland: a catalytic life.

Professor W. Wallace Cleland, the architect of modern steady-state enzyme kinetics, died on March 6, 2013, from injuries sustained in a fall outside of his home. He will be most remembered for giving the enzyme community Ping-Pong kinetics and the invention of dithiothreitol (DTT). He pioneered the utilization of heavy atom isotope effects for the elucidation of the chemical mechanisms of enzyme-catalyzed reactions. His favorite research journal was Biochemistry, in which he published more than 135 papers beginning in 1964 with the disclosure of DTT.

In situ measurements of brain tissue hemoglobin saturation and blood volume by reflectance spectrophotometry in the visible spectrum.

Before the development of near-infrared spectroscopy (NIRS) for monitoring of hemoglobin and cytochromes in situ, the Jobsis laboratory designed a visible light reflectance spectrophotometer. The method was not as useful for cytochrome oxidase measurements, which stimulated the search for a better method that culminated in NIRS. Visible light reflectance spectrophotomery was, however, usefully applied in several experimental applications, such as the study of brain capillary hemoglobin saturation during changes in inspired gas mixtures in awake and anesthetized animals, and to record transient increases in total hemoglobin (blood volume) after local neuronal activation by direct cortical electrical stimulation, demonstrating a response that is fundamental to functional magnetic resonance imaging blood oxygen level-dependent methods. A third application of the instrumentation was for brain capillary red cell mean transit time analysis, estimated by recording the passage of a red cell-free bolus through the cerebral cortical optical monitoring field. Taken together with his previous application of fluorescence detection of nicotinamide adenine dinucleotide, the visible and near-infrared spectroscopy demonstrate that Frans Jobsis was a pioneer in the application of optical techniques to the study of intact organs in situ. These methods have been used to illuminate the basic function of the cerebrovascular and metabolic pathways in both physiological and pathological conditions.

How hydrides misled chemists.

Hydrogen-containing molecules are simple enough to be attractive subjects in experimental diffraction and spectroscopic studies and in quantum computations. Yet, the inferences about molecular structure and force fields originally drawn from studies of these subjects were significantly flawed. In recent developments the original models of structure invoked, such as hybridization, have been superseded. The reasons for this are briefly reviewed. What has emerged to account for molecular geometry, prevailing even over the popular VSEPR theory, is a model of geminal nonbonded interactions.

Spectroscopy of succinate dehydrogenases, a historical perspective.

An attempt is made to retrace, from personal experience, the discovery of redox-reactive non-heme iron in living matter, which turned out to occur in the form of iron-sulfur (Fe-S) clusters, and then to recount the immediate application of this knowledge in exploring the composition of the mitochondrial respiratory chain, and in the rather detailed description of the workings of its components and, for the purposes of the present volume, of succinate dehydrogenase. The relationship of these events to the general status of technology and the available methodology and instrumentation is considered in some detail, with the conclusion that there scarcely was a way that these discoveries could have been made earlier. It is then shown how methods, techniques and interpretations of results were developed and evolved during the applications that were made to a complex problem such as that of the composition, structure and functioning of succinate dehydrogenase. A tabulation of the most significant events--concerning specifically spectroscopy and its interpretations--in this development is given up to the year 2000.

Albumin standards and the measurement of serum albumin with bromcresol green. 1971.

A rapid and reliable method for measuring serum albumin employing bromcresol green is described. The addition of albumin to a solution of bromcresol green in a 0.075 M succinate buffer pH 4.20 results in an increase in absorbance at 628 nm. The absorbance-concentration relationship is linear for samples containing up to 6 g/dl albumin. Bilirubin, moderate lipemia, and salicylate do not interfere with the analysis. The use of nonionic surfactant (Brij-35) reduces the absorbance of the blank, prevents turbidity and provides linearity. The results by this method agree very well with those obtained by electrophoresis and salt fractionation. The method is simple, it has excellent precision and the reagents are stable. A protein standard is introduced which can be employed for both the total serum proteins and albumin determinations.

History of blood gas analysis. VI. Oximetry.

Oximetry, the measurement of hemoglobin oxygen saturation in either blood or tissue, depends on the Lambert-Beer relationship between light transmission and optical density. Shortly after Bunsen and Kirchhoff invented the spectrometer in 1860, the oxygen transport function of hemoglobin was demonstrated by Stokes and Hoppe-Seyler, who showed color changes produced by aeration of hemoglobin solutions. In 1932 in Göttingen, Germany, Nicolai optically recorded the in vivo oxygen consumption of a hand after circulatory occlusion. Kramer showed that the Lambert-Beer law applied to hemoglobin solutions and approximately to whole blood, and measured saturation by the transmission of red light through unopened arteries. Matthes in Leipzig, Germany, built the first apparatus to measure ear oxygen saturation and introduced a second wavelength (green or infrared) insensitive to saturation to compensate for blood volume and tissue pigments. Millikan built a light-weight ear "oximeter" during World War II to train pilots for military aviation. Wood added a pneumatic cuff to obtain a bloodless zero. Brinkman and Zijlstra in Groningen, The Netherlands, showed that red light reflected from the forehead could be used to measure oxygen saturation. Zijlstra initiated cuvette and catheter reflection oximetry. Instrumentation Laboratory used multiple wavelengths to measure blood carboxyhemoglobin and methemoglobin is cuvette oximeters. Shaw devised an eight-wavelength ear oximeter. Nakajima and co-workers invented the pulse oximeter, which avoids the need for calibration with only two wavelengths by responding only to the pulsatile changes in transmitted red and infrared light. Lübbers developed catheter tip and cuvette fiberoptic sensors for oxygen tension, carbon dioxide tension, and pH.