Effect of L-methionine supplementation on plasma homocysteine and other free amino acids: a placebo-controlled double-blind cross-over study.

OBJECTIVE: The essential amino acid L-methionine is a potential compound in the prophylaxis of recurrent or relapsing urinary tract infection due to acidification of urine. As an intermediate of L-methionine metabolism, homocysteine is formed. The objective was to study the metabolism of L-methionine and homocysteine, and to assess whether there are differences between patients with chronic urinary tract infection and healthy control subjects. DESIGN: A randomized placebo-controlled double-blind intervention study with cross-over design. SETTING: Department of Nutritional Physiology, Institute of Nutrition in cooperation with the Department of Internal Medicine III, Friedrich Schiller University of Jena, Germany. SUBJECTS: Eight female patients with chronic urinary tract infection and 12 healthy women (controls). INTERVENTIONS: After a methionine-loading test, the volunteers received 500 mg L-methionine or a placebo three times daily for 4 weeks. MAIN OUTCOME MEASURES: Serum and urinary concentrations of methionine, homocysteine, cystathionine, cystine, serine, glycine and serum concentrations of vitamin B12, B6 and the state of folate. RESULTS: Homocysteine plasma concentrations increased from 9.4+/-2.7 micromol/l (patients) and 8.9+/-1.8 micromol/l (controls) in the placebo period to 11.2+/-4.1 micromol/l (P=0.031) and 11.0+/-2.3 micromol/l (P=0.000), respectively, during L-methionine supplementation. There were significant increases in serum methionine (53.6+/-22.0 micromol/l; P=0.003; n=20) and cystathionine (0.62+/-0.30 micromol/l; P=0.000; n=20) concentrations compared with the placebo period (33.0+/-12.0 and 0.30+/-0.10 micromol/l; n=20). Simultaneously, renal excretion of methionine and homocysteine was significantly higher during L-methionine intake. CONCLUSIONS: Despite an adequate vitamin status, the supplementation of 1500 mg of L-methionine daily significantly increases homocysteine plasma concentrations by an average of 2.0 micromol/l in patients and in control subjects. An optimal vitamin supplementation, especially with folate, might prevent such an increase.

Receptor-ligand interactions studied with homogeneous fluorescence-based assays suitable for miniaturized screening.

Cell membrane receptors play a central role in controlling cellular functions, making them the target of drugs for a wide variety of diseases. This report describes how a recently developed method, fluorescence intensity distribution analysis (FIDA), can be used to develop homogeneous, nonradioactive high throughput screening assays for membrane receptors. With FIDA, free ligand and ligand accumulated on receptor-bearing membrane vesicles can be distinguished on the basis of their particle brightness. This allows the concentration of both bound and free ligand to be determined reliably from a single measurement, without any separation. We demonstrate that ligand affinity, receptor expression level, and potency of inhibitors can be determined using the epidermal growth factor and beta(2)-adrenergic receptors as model systems. Highly focused confocal optics enable single-molecule sensitivity, and sample volumes can thus be reduced to 1 microl without affecting the quality of the fluorescence signal. Our results demonstrate that FIDA is an ideal method for membrane receptor assays offering substantial benefits for assay development and high throughput pharmaceutical screening.

[Enzootic calcinosis in 16 cows from 6 dairy farms in Unterengadin]. / Enzootische Kalzinose bei 16 Kühen aus 6 Milchviehbetrieben im Unterengadin.

Enzootic calcinosis was diagnosed in 16 cows, which originated from 6 dairy farms located in the Unterengadin valley in Switzerland between 1,402 and 1,650 metres above sea level. During the winter, the cows were fed haylage, which contained up to 50 per cent golden oat-grass (Trisetum flavescens), and hay and grain. The first symptoms of enzootic calcinosis were noted in March 1998, when some of the cows developed locomotor abnormalities. Additional cases were diagnosed during the following three months. The most important clinical findings were decreased milk production, weight loss, frequent recumbency, difficulty in rising, kneeling while rising and feeding, stilted gait, arched back, shifting weight from one foot to another and reluctance to remain standing after being roused. Of the 16 cows, 5 had elevated concentrations of serum calcium and 10 had decreased concentrations of serum magnesium. All cows had normal or decreased concentrations of serum phosphorus, because the feeding of haylage containing golden oat-grass had been discontinued two months previously. In all of the cows, the most important postmortem findings were severe calcification of the aorta, iliac artery, brachiocephalic trunk and pulmonary artery.

A prospective study of the risk of transfusion-acquired viral infections.

The risk of transfusion-transmitted viral infections may be estimated by several methods, but only prospective studies of transfusion recipients can directly measure the incidence, with associated 95% upper confidence bound, of these infections. From 1989 through 1995, 764 recipients of allogeneic or autologous red blood cell transfusions were enrolled; 486 (64%) provided both pretransfusion and 6-month follow-up specimens. Both specimens were tested for anti-HBc, anti-HCV, anti-HTLV-I and anti-HIV, with appropriate confirmatory testing. Thirty-nine (8.0%) subjects had seroprevalent anti-HBc, 19 (3.9%) subjects had seroprevalent anti-HCV, three (0.6%) subjects had seroprevalent anti-HTLV-I/II, and one (0.2%) subject had seroprevalent anti-HIV. There were no seroconversions for any agent among the 34 patients who received only autologous blood, and no confirmed seroconversions for anti-HTLV-I or anti-HIV among all subjects. There were three seroconversions for anti-HBc (incidence 1.04 x 10(-3); 95% confidence interval (CI) 2.15 x 10(-4), 3.05 x 10(-3) per allogeneic unit transfused), and two confirmed seroconversions for HCV (incidence 6.94 x 10(-4); 95% CI 8.34 x 10(-5), 2.51 x 10(-3) per allogeneic unit transfused). One of the two anti-HCV seroconversions occurred in March 1994, after the institution of HCV EIA 2.0 screening of donated blood. Transfusion-associated seroconversions to hepatitis B and C markers were observed at low rates in the early 1990s despite testing donors for markers of both viruses, whereas seroconversions to HTLV-I or HIV were less than 1.04 x 10(3) per allogeneic unit transfused, based upon the upper 95% confidence interval of the zero incidence in this study.

[Recent application methods of high dose-rate brachytherapy in head and neck tumors]. / Neuere Applikationsmethoden der Brachytherapie mit hoher Dosisleistung bei Kopf-Hals-Tumoren.

A striking development has been made in intracavitary and interstitial brachytherapy of head and neck tumors by the use of very active and very small ray emitters and, last but not least, by the step movement of the source developed 1974 in Essen and its computer optimization. The present study mentions 22 different sites of the head and neck area which have been treated by 31 different intracavitary or interstitial techniques, frequently under general anaesthesia. All application methods used here follow uniform rules for the dosage of the single application. Furthermore, generally accepted rules can be defined for the contra-indications of brachytherapy which, according to our experiences, should be strictly observed. The observance of these rules as well as the use of a radiobiological gap were among the reasons for the complete recovery of some cases of local recurrence which could be no longer treated by other methods.