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P J Geiger

Publications and source records attributed to P J Geiger.

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Radioimmunoassay data handling and calculations with a graphics-statistics computer program.

This paper discusses the application of the data handling-graphics-statistics program Stata (Computing Resources Center, Santa Monica, CA) to radioimmunoassay. We have found that this program is more powerful and easier to use than a spreadsheet for analyzing various kinds of laboratory data generated from chromatography, radiolabeling experiments, enzyme-linked immunosorbent assays, and radioimmunoassays, to name several examples. Data from a radioimmunoassay procedure, originally analyzed using a spreadsheet, Lotus 1-2-3, have been processed with Stata. Simple programs (batch files) have been devised for computations and graphics. The original data and a comparison of results are presented.

Radioimmunoassay

Lack of hormonal stimulation of pyridoxine metabolism in isolated rat hepatocytes.

Isolated hepatocytes obtained from Sprague-Dawley rats (145-175 g) were incubated for 15 min at 30 degrees C in Krebs-Henseleit bicarbonate buffer, pH 7.4, containing 0.5 mM concentration of each of the 20 natural amino acids and either 4.5 or 23 microM [U-14C]pyridoxine. Pyridoxine, pyridoxal, pyridoxal phosphate, and pyridoxic acid separated by an anion-exchange chromatographic technique were quantified using a phosphate analyzer and a liquid scintillation counter. The conversion of [U-14C]pyridoxine to its metabolites was more than doubled by increasing the amount of pyridoxine (4.5 to 23 microM) in the incubation medium. Insulin (10 mU/ml), glucagon (1 nM), or epinephrine (10 microM) did not have any significant effect on the conversion of [14C]-pyridoxine to pyridoxal, pyridoxal phosphate, or pyridoxic acid. Our earlier observations of a large decrease in serum pyridoxal phosphate in the diabetic rat cannot be explained by any direct hormonal effects on pyridoxine metabolism.

Animals

Prevalence of Corynebacteria in diabetic foot infections.

OBJECTIVE: Microbiological flora of diabetic foot infections are usually polymicrobial and frequently include bacteria of the Corynebacterium sp. (diphtheroids). The purpose of this study was to determine the prevalence of these bacteria in both deep and superficial cultures in diabetic patients with foot infections. RESEARCH DESIGN AND METHODS: The charts of 50 patients of successive admissions to the Orthopedic-Diabetes Service at our hospital were reviewed to obtain the following data: age, sex, ethnic origin, method of treatment of diabetes, blood glucose level, prior antibiotics, and reports of cultures taken from bedside and intraoperative sites. Data were analyzed to compare the prevalence of diphtheroids in reliable versus nonreliable cultures and the influence of other parameters on the presence of these organisms. RESULTS: Fourteen of 19 (74%) of the intraoperative specimens grew diphtheroids compared with 25 of 65 (39%) of the bedside cultures, a highly significant difference. In addition, there was a somewhat greater occurrence of diphtheroids in women compared with men. The likelihood that contamination is the cause for the presence of diphtheroids is highly unlikely, because one arm of the study included cultures derived from deep tissue at the time of the surgical procedure (i.e., the intraoperative cultures). Cultures always grew at least one other organism in addition to the diphtheroid. CONCLUSIONS: Corynebacteria, commonly known as diphtheroids, are present as a part of the polymicrobial flora in a large percentage of diabetic patients with foot infections. Because the diphtheroids were identified in culture material taken in the operating room or at the time of incision and drainage in a higher percentage of patients than in specimens from superficial cultures, it is highly unlikely that they are contaminants.

Corynebacterium

Extracellular phosphate requirement for insulin action on isolated rat hepatocytes.

Isolated rat hepatocytes were prepared in KHB buffer, pH 7.4; were centrifuged and washed twice in KHB buffer containing various amounts of phosphate and calcium; and were incubated at 30 degrees in the presence of tracer [2,3-14C]succinate and a 0.5 mM concentration of each of the 20 natural amino acids. Hepatocytes washed and incubated in KHB buffer containing less than 0.1 mM phosphate failed to show any insulin stimulation of [2,3-14C]succinate oxidation or protein incorporation of tracer carbons. The absence or presence of extracellular phosphate did not alter the specific activity of 32P-adenine nucleotides; they remained the same in the presence or absence of insulin. The maximal insulin stimulatory effect on succinate oxidation and tracer incorporation into protein was observed in the presence of 1.18 mM phosphate and 1.9 mM calcium ion. The lack of external phosphate did not prevent the stimulation of succinate oxidation by either glucagon on epinephrine, whereas removal of calcium from the medium abolished their hormonal effects. The lack of medium calcium also prevented the insulin stimulation of succinate oxidation and protein synthesis. Our data indicate that a diminished insulin responsiveness in hypophosphatemic patients may be due to the insensitivity of mitochondria to insulin in the hypophosphatemic state.

Animals

The relation of diabetic control to in vivo pH of soft tissue abscesses.

It has been shown that induced soft tissue abscesses have a lower intra-abscess pH in the uncontrolled diabetic host than in the nondiabetic control. These differences were felt to be secondary to alterations in white cell metabolism. The current study compares the intra-abscess pH in three groups of mice: (I) nondiabetic, (II) untreated diabetic, and (III) insulin-treated diabetic. Diabetes was induced with streptozotocin in male white mice. The bacteria used to induce the abscesses were a combination of B. fragilis and Enterococcus. The blood glucose values of groups I, II, and III were 189 mg% (+/- 20.3), 256 mg% (+/- 121.9), and 712.8 mg% (+/- 169.7), respectively. None of the animals were ketotic, and peritoneal pH (reflecting systemic pH) showed no significant differences between groups. There were no significant differences in colony counts between any groups. The intra-abscess pH values of groups I, II, and III were 6.97 (+/- 0.26), 6.85 (+/- 0.41), and 6.08 (+/- 0.70). The differences in intra-abscess pH and blood glucose levels were all significantly different from each other when all three groups were compared. The insulin-treated mice tended to return to normality but had the widest spread of values. Since a decrease in intra-abscess pH has been felt to be a reflection of white cell activity, our studies may be the first to demonstrate an in vivo effect of insulin on white cell activity.

Abscess