Automated method for L-carnitine determination

Because of renewed interest in a possible connection between carnitine, lipid disorders, and myopathy, an automated method of analysis is desirable. Deproteinization of serum by use of membrane filter cones and automated assay with a bichromatic analyzer (the ABA-100) substantially increases efficiency without sacrificing the specificity and accuracy of the original manual enzymatic method. The described procedure allows for analysis of 80 speciments a day and is thus suitable for screening of selected populations. Normal values found in blood sera of adults were in the range of 25.0-73.8 mu mol/liter and the method has sufficient sensitivity to accurately measure concentrations as small as 10 mu mol/liter.     

Decreased antidiuretic response in diabetes insipidus during prolonged treatment with carbamazepine presumably due to enzym-induction

Carbamazepine (CA)-induced antidiuresis and fluorescence in urinary cortisol assay was compared during short-term and prolonged treatment in 3 patients with pituitary diabetes insipidus. The fluorescence induced by excreted CA-metabolite(s) (probably CA-10-11 epoxide and its derivatives) was used for the rough clinical estimation of the rate of metabolic elimination of the drug. A marked decrease was observed both in the antidiuretic response induced by CA and excretion of CA metabolite(s) during prolonged treatment - with a fixed dose of CA. These findings suggest that diminishing antidiuretic effectiveness might be due to auto-induction of CA. Auto-induction of CA may explain the delay in improvement of renal concentrating operation in patients with DI during long-term treatment with fixed doses of CA. Another clinical consequence of this mechanism may be the prevention of the development of water intoxication in neuro-psychiatric patients undergoing chronic treatment with CA.     

Nanosecond-pulse fluorimetry of wheat-germ agglutinin (lectin)

Nanosecond-pulse fluorimetry of wheat germ agglutinin is analyzed as a function of both excitation and emission wavelengths. When excited at 280 nm, wheat germ agglutinin fluorescence exhibited three lifetimes: one corresponding to the tyrosine residues as a whole and two others corresponding to the tryptophyl emission. The tyrosine contribution to the emission spectrum deduced from this method was in good agreement with that reported previously in steady-state fluorescence experiments [Privat, J.P. and Monsigny, M. (1975) Eur. J. Biochem. 60, 555-567]. The fluorescence decay of each tryptophan residue was not a single exponential function when wheat germ agglutinin was excited at 295 nm. This could be related to the microenvironment of the indole chromophores in the protein. The comparison of the quantum yield and of average lifetime showed that some tryptophan residues were completely quenched. Energy transfer from tyrosines to tryptophan residues previously detected in steady-state fluorescence was also revealed by fluorescence decay measurements. Comparison of both methods showed that an important part of transfers occurred with a very fast rate equal to or greater than 10(10) s-1. Both lifetimes and the ratio of the short and the long-lived component were found dependent on tri-N-acetylchitotriose binding.     

Release of renal prostaglandins during endotoxin-induced hypotension

The appearance of prostaglandins in dog's blood during endotoxin-induced hypotension was studied by use of the dialysis modification of the blood bathed organ technique. An increase in prostaglandins, mainly E2 and F2alpha was found in renal venous blood, whereas no such increase was seen in blood from the abdominal aorta, the inferior vena cava or the femoral vein. Three possible trigger mechanisms for this increase i.e. hypotension, reduced flow and reflexogenic sympathetic stimulation, have been investigated. It is suggested that, in addition to these three factors, circulating hormones such as noradrenaline, angiotensin or bradykinin, play a role in this release mechanism. Administration of indomethacin produced a restoration of the systemic blood pressure to its pre-endotoxin value; concomitantly a disappearance of the prostaglandins from the circulation was observed. It is concluded that prostaglandins contribute to the hypotension induced by endotoxin. Whether they are beneficial or detrimental remains to be resolved.     

Triglyceride lipases in acute hepatitis

Two triglyceride lipases in postheparin plasma, the lipoprotein lipase (LPL) and the hepatic triglyceride lipase (H-TGL) were separated by heparin-sepharose affinity chromatography and studied in controls and patients during the course of acute hepatitis. All three patients had increased content of triglycerides in the low density lipoproteins, and two of them had hypertriglyceridemia. Low activities of both lipases were found in the acute stage of the disease in all three patients. Concomitantly one of the patients had absolute low lecithin: cholesterol acyltransferase (LCAT) activity, and in the two other patients a relative LCAT deficiency was present. The increased content of triglycerides in LDL that may be found in liver disease, may not only be due to low H-TGL and LPL, but also LCAT deficiency.     

Study on the relationship between plasma prolactin levels and androgen metabolism in man

The purpose of the study was to investigate the effect of hyperprolactinemia on the metabolism of androgens in man. A group of 6 normal men was treated for 4 consecutive days, on separate periods, with Sulpiride which is known to raise plasma prolactin (PRL) concentration. The effect of the treatment on plasma steroids was verified in basal conditions and under stimulation by HCG. In the controls, a parallel rise in testosterone (T) and dihydrotestosterone (DHT) was observed in response to HCG stimulation. In experimental hyperprolactinemia, the rise in T in response to HCG, similar to that of the controls, was accompanied by a markedly diminished rise in DHT. Similar results were observed in a patient with hyperprolactinemia following apparent accidental section of the pituitary stalk. These data demonstrate the interference of increased levels of PRL in the metabolism of testosterone into the active DHT form by 5alpha-reductase. They suggest that this mechanism could possibly impair the gonadal function in man.     

Rheographic assessment of cerebral blood volume and correlations with changes in intracranial pressure

The authors present a four electrode rheographic technique for simiquantitative continuous measurement of CBV, by examining changes in cerebral tissue impedance induced by changes in blood content of the brain parenchyma. Correlation between the data obtained and ICP measurements permits conclusions regarding the behaviour of the cerebral blood bed. pCO2 regulates CBV, and the phenomenon is reversible according to a hystereris shaped time course in acute experiments. The rebound of ICP after sudden injections of CSF into the cisterna magna is of a vasogenic nature. Changes in CSF volume induce contrary changes in CBV. Pharmacological effects can be studied. The validity of the method is discussed.     

Subendocardial blood flow in children with congenital aortic stenosis

Subendocardial blood flow may be estimated from the ratio of flow to the subendocardium to myocardial oxygen consumption. The first may be estimated from the diastolic pressure time index (area between aortic and left ventricular (LV) pressure during diastole) and the latter by the tension time index (integral of LV pressure during systolic ejection). Subendocardial flow index (SEFI) averaged 1.27 (0.96-1.78) in 13 children with normal aortic valves. SEFI averaged 0.88 (0.43-1.65) in asymptomatic children with congenital aortic stenosis and was never greater than 0.9 in symptomatic children. Aortic valve area and systolic pressure difference did not correlate well with symptoms. SEFI and aortic valve area increased in 26 of 28 patients after surgery. However, 23 of 28 had varying degrees of aortic regurgitation following valvotomy. Since calculation of SEFI is not affected by aortic regurgitation, it would appear to be a more useful measure of surgical success than aortic valve area.     

Occlusion pressures in men rebreathing CO2 under methoxyflurane anesthesia

The effect of general anesthesia on control of breathing was studied by CO2 rebreathing and occlusion pressure measurements in six normal human subjects under methoxyflurane anesthesia. CO2 was found to increase the amplitude of the occlusion pressure wave without changing its shape, so that CO2 responses in terms of the occlusion pressure developed 100 ms after the onset of inspiration (Po/0.1) gave results equivalent to the responses in terms of Po/1.o or any other parameter of the pressure wave. Methoxyflurane depressed the ventilatory response to CO2 but not the occlusion pressure response, implying that the most important action of the anesthetic was to increase the effective elastance of the respiratory system rather than to depress the respiratory centers. The elastance was further increased by CO2, and this mechanical change had the effect of shifting the "apneic threshold" extrapolated from the ventilatory response curve to a lower PAco2. Frequency of breathing, inspiratory and expiratory times were not altered by CO2 in anesthetized subjects.