Microhetero;geneity and polymorphism of human serum thryroxine-binding globulin. Study by isoelectric focusing and radioprint immunofixation.

A new technique of radioprint immunofixation following isoelectric focusing was applied to the study of microheterogeneity of thyroxine-binding globulin without purification, in individual sera; this protein appeared to give rise to at least seven bands with isoelectric points in the range of pI 4.2--5.0. Furthermore, two different distributions of the intensity of these bands were found in the sera investigated, suggesting the existence of thyroxine-binding globulin polymorphism in man.

Role of sialic acid in the microheterogeneity of serum thyroxine-binding globulin. Study by two-dimensional isoelectric focusing.

A new technique combining a neuraminidase treatment of thyroxine-binding globulin after serum isoelectric focusing and a second-dimensional isoelectric focusing was developed to study the role of sialic acid in the microheterogeneity of native thyroxine-binding globulin. By showing the change of PI occasioned by the desialylation for each of the bands constituting the thyroxine-binding globulin pattern separately, this procedure clearly demonstrated that the microheterogeneity of the native protein could not be imputed to the varying sialic acid content of the bands only. We suggest that at least three molecules of thyroxine-binding globulin, with probably slight differences in their amino acid composition, are present in the serum, and that different degrees of sialylation ensure greater microheterogeneity of this protein.

Radioimmunofixation of human ferritin following serum isoelectric focusing.

Radioimmunofixation of human ferritin following isoelectric focusing of serum was developed to study the microheterogeneity of this protein in native serum without previous purification or concentration. This method requires only 2-10 microliter of serum and can be used with levels of ferritin as low as 10 micrograms/l. In this way, the extensive microheterogeneity of this protein was revealed, since in some cases it produced as many as 35 bands with isoelectric points in a pH range of 4.95-5.9. Very different isoelectric focusing patterns (spectrotypes) of ferritin were observed during the investigation of pathological sera. The high sensitivity of this technique makes it useful for the investigation of serum ferritin in diseases involving modifications of the metabolism of this protein.

Identification of atypical strains of Staphylococcus epidermidis by use of molecular tools.

Four atypical coagulase-negative staphylococcal (CNS) isolates from clinical sources were compared with Staphylococcus epidermidis strains by ribotyping. The ribotypes of the four strains shared close rDNA restriction profiles with those of the S. epidermidis strains used. The DNA sequence encoding 16S rRNA demonstrated 99.9% homology with S. epidermidis. S1 nuclease experiments showed that these atypical strains formed a homogeneous genomic group. DNA-DNA homologies between the S. epidermidis type strain CCM 2124 and the four CNS isolates ranged from 70 to 89%. The guanine-plus-cytosine content of the deoxyribonucleic acid of the four strains ranged from 31 to 32 mol%.

Effect of oral administration of lipids with 67% medium chain triglycerides on glucose homeostasis in preterm neonates.

Since hypoglycemic responses to medium chain triglycerides (MCT) have been reported in adults we studied the effect of an acute oral load of lipids (2,8 g/kg) with 67% MCT on glucose homeostasis in 21 preterm infants in comparison to 14 age-matched control preterm infants. A hyperglycemic response from (mean +/- SEM) 57 +/- 1.1 to 74 +/- 2.5 at 30 min (p less than 0.01) and to 80.5 +/- 2.5 mg/dl at 60 min (p less than 0.01) was observed after administration of the lipids whereas no change in plasma glucose concentration was observed in the control group. After administration of the lipids there was no change in the concentration of insulin and glucagon in plasma. An intravenous glucose tolerance test (1 g/kg) was similar in the control group and 60 min after administration of the lipids. After administration of the lipids free fatty acid concentration remained unchanged while a significant decrease from 304 +/- 56 to 199 +/- 28 muEq/l was observed in 60 min in the control group. At 60 min beta-hydroxybutyrate concentration was higher after lipid administration (630 +/- 86 mumol/l) than in the control group (436 +/- 66 mumol/l) (p less than 0.05). A more rapid decrease in blood lactate concentration was found after lipid administration than in the control group while no change in plasma alanine concentration was observed in either groups. In five additional preterm infants, plasma glucose concentration increased from 56 +/- 0.6 to 75 +/- 0.9 mg/dl (p less than 0.01) 60 minutes after gastric administration of glycerol (0.3 g/kg). These data show that in preterm infants, a lipid load with 67% TCM produces a hyperglycemic response through gluconeogenesis without changing the peripheral rate of glucose disappearance.

The sodium pump of erythrocytes from patients with Duchenne muscular dystrophy: effect of ouabain on the active sodium flux and on (Na+, K+)ATPase.

Active ion exchanges across erythrocyte membrane from patients with Duchenne muscular dystrophy were studied by two methods: --erythrocyte ghosts (Na+,K+)ATPase activity and its sensibility to ouabain, using an enzymatic method; --active sodium outflux and its sensibility to ouabain using a radioisotopic method. (Na+,K+)ATPase activity and active sodium outflux were decreased in dystrophic patients compared to normal sex- and age-matched controls. Enzymatic activity and active sodium outflux were inhibited by ouabain 10(-4) M in patients and in controls. Inhibition of the active sodium outflux was lower in patients than in controls. The results are discussed in light of the literature.

Serum ferritin spectrotypes in patients with heterozygous beta-thalassaemia.

Serum ferritin spectrotypes from patients heterozygous for beta-thalassaemia were determined after agarose isoelectric focusing followed by radio-immunofixation with anti-ferritin antibody. Multivariate analysis demonstrated a specific spectrotype for heterozygous beta-thalassaemia. This spectrotype was shown to be different from those in hereditary spherocytosis and idiopathic haemochromatosis. Statistical discrimination reached 100% of well-classified patients between these pathological conditions.

Assessment of proportional growth of very low birth weight infants fed banked human milk.

Twenty appropriate (mean +/- S.D., gestational age (AGA): 29.9 +/- 1.5 weeks) and 15 small (GA: 34.6 +/- 2.4 weeks) for gestational age (SGA) very low birth weight infants fed banked mature human milk were studied until term for anthropometric parameters: midarm (MAC), chest (CC), head (HC) circumferences, triceps (TSKF) and subscapular (SSKF) skinfold thickness recorded at 15 and 60 s, dynamic skinfold (delta % SKF), muscle (AMA) and fat (AFA) areas, weight and length. In AGA infants, all the parameters at term were significantly lower in extrauterine (EUL) that in intrauterine life (IUL). At term the relative proportion of AFA to total arm area was increased in EUL compared to IUL both in AGA (25.87 +/- 3.8 vs. 23.26 +/- 1.27% respectively, P less than 0.01) and in SGA infants (21.89 +/- 4.63 vs. 18.81 +/- 3.9 respectively, P less than 0.05). SGA infants showed a similar growth in EUL compared to IUL, and a significantly lower AMA and AFA than in AGA infants in EUL. Although HC was in both infants below the 10th centile at term, the ratio weight/HC2 suggests a relative preservation of head growth in EUL compared to IUL (AGA: 20.72 less than 0.87 vs. 22.65 +/- 1.46 respectively, P less than 0.001; SGA; 20.82 +/- 1.16 vs. 21.62 +/- 1.86 respectively, NS). Delta %SKF were negatively correlated with post-conceptional age suggesting a loss of extracellular water in AGA (delta %TSKF: r = -0.287, P less than 0.02) and in SGA infants (delta %TSKF: r = -0.301, P less than 0.02; delta %SSKF: r = -0.316, P less than 0.02). An intrauterine model of discrimination between AGA and SGA infants does not apply to EUL. An equation was established in SGA infants with the best discriminant parameters giving a predictive post-conceptional age: post-conceptual age (PCA) (weeks) = 0.276 HC (cm) + 0.723 CC (cm) - 0.122 MAC (cm) + 0.5 TSKF (mm) + 10.173, (r = 0.867, P less than 0.001) allowing a clear discrimination between AGA and SGA infants. These results suggest that infants show quite different growth patterns between IUL and EUL both for AGA and SGA infants.

Anthropometric assessment of nutritional status in newborn infants. Discriminative value of mid arm circumference and of skinfold thickness.

74 appropriate-for-gestational age (AGA) and 22 small-for-gestational age (SGA) caucasian infants were studied for anthropometric parameters: mid arm circumference (MAC), triceps and subscapular skinfold thickness (TSKF and SSKF) recorded at 15 and 60 s, chest circumference (cc), head circumference, birth weight and length. MAC is highly correlated with birth weight either in AGA (r = 0.936; P less than 0.001) or in SGA infants (r = 0.860; P less than 0.001). MAC is also correlated with gestational age in AGA (r = 0.850; P less than 0.001) and SGA infants (r = 0.76; P less than 0.001). Similar correlations were found between TSKF, SSKF and birth weight or gestational age. Arm muscle and fat areas are also positively correlated with birth weight and gestational age, in AGA and SGA infants. A multiple regression analysis of our data allowed a classification of the best discriminant anthropometric parameters between AGA and SGA infants. MAC, SSKF15, SSKF60 and chest circumference were selected. An equation was established in AGA infants with these four parameters giving a predictive gestational age: gestational age (weeks) = 1.216 MAC (cm)-3.588 SSKF15 (mm) + 0.263 CC (cm) + 17.9. The ratio of predicted gestational age to the real gestational age was 1.0 +/- 0.044 in AGA versus 0.896 +/- 0.034 in SGA infants. Our data suggest that MAC and SSKF provide a simple measure of body composition of neonates and a useful tool for determining the degree of maturity of a newborn independent of birth weight.

Transrectal high-intensity focused ultrasound: minimally invasive therapy of localized prostate cancer.

BACKGROUND AND PURPOSE: Criteria for determining the durability of the response to transrectal high-intensity focused ultrasound (HIFU) ablation of prostate cancer have been established by calculating progression-free probability. PATIENTS AND METHODS: A series of 82 patients (mean age 71 +/- 5.7 years) with biopsy-proven localized (stage T1-T2) cancer who were not suitable candidates for radical surgery underwent transfectal HIFU ablation with the Ablatherm machine. The mean follow-up was 17.6 months (range 3-68 months). The mean serum prostate specific antigen (PSA) value and mean prostate volume were 8.11 +/- 4.64 ng/mL and 34.9 +/- 17.4 cm3, respectively. Progression was rigidly defined as any positive biopsy result, regardless of PSA concentration, or three successive PSA increases for patients with a negative biopsy (PSA velocity > or = 0.75). Times to specific events (positive biopsy and PSA elevation) were analyzed with the Kaplan-Meier survival method. RESULTS: Overall, 62% of the patients exhibited no evidence of disease progression 60 months after transrectal HIFU ablation. In particular, the disease-free rate was 68% for the moderate-risk group of 50 patients (PSA < 15.0 ng/mL, Gleason sum < 8, prostate volume < 40 cm3, and number of positive biopsies < 5). For the low-risk group of 32 patients (PSA < 10 ng/mL and Gleason sum < 7), the disease-free survival rate was 83%. CONCLUSION: Transrectal HIFU prostate ablation is an effective therapeutic alternative for patients with localized prostatic adenocarcinoma.

Non-linear patterns of growth in very low birth weight infants.

The deviations of daily weight, weekly length and head circumference from linear growth were analyzed in 87 very low birth weight infants. The deviations exhibited a highly significant polynomial oscillation not only for weight, but also for length and head circumference. The weight amplitudes were larger for males than for females. They were also larger in infants appropriate for gestational age than in small-for-date infants. The difference with trophicity may be related to the process of adjustment of catch-up in small-for-date infants. However, the mechanisms of these oscillatory deviations could also be explained by clinical events, method of feeding, or homeostatic regulation. Further studies are required to elucidate the role of the different factors. Am. J. Hum. Biol. 10:637-646, 1998. © 1998 Wiley-Liss, Inc.

[Course of the radionuclide left ventricular ejection fraction at rest and during exercise in surgically treated asymptomatic chronic aortic insufficiency]. / Evolution de la fraction d'éjection radio-isotopique ventriculaire gauche au repos et á l'effort dans l'insuffisance aortique chronique asymptomatique opérée.

In order to preserve left ventricular (LV) function, aortic valve replacement may be contemplated in asymptomatic patients with aortic regurgitation when LV dilatation and dysfunction are not too advanced. Our study involved 10 asymptomatic patients with severe, isolated and pure aortic regurgitation. Before, and 6 months after the operation, the LV ejection fraction (LVEF) was measured at rest and during exercise on an ergometric bicycle by radionuclide angiography (multigated technique). Mean preoperative values were: age 52 +/- 14 years; cardiothoracic ratio 0.55 +/- 0.04; end-diastolic LV diameter 69 +/- 9 mm; end systolic LV diameter 47 +/- 7 mm; LV fibre shortening fraction 0.31 +/- 0.03; LVEF 0.55 +/- 0.10 at rest and 0.41 +/- 0.13 at exercise. After surgery, the cardiothoracic ratio value (0.51 +/- 0.03) and the LVEF value at rest (0.60 +/- 0.07) were not significantly different from the corresponding preoperative values, but the LVEF value during exercise was significantly increased (0.58 +/- 0.11, p less than 0.001). Among the 9 patients who before surgery showed a fall in LVEF at exercise, after surgery 5 had a rise (group B) and 4 had a fall (group A) in LVEF at exercise. Before surgery, group A patients had greater LV diameters than group B patients: end-diastolic diameter 76 +/- 6 mm vs 63 +/- 9 mm; end-systolic diameter 53 +/- 4 mm vs 43 +/- 7 mm (p = 0.07). These diameters were the only variables that predicted the postoperative changes in LVEF at exercise.(ABSTRACT TRUNCATED AT 250 WORDS)

[Left ventricular function in chronic asymptomatic aortic insufficiency. Angioscintographic study]. / Fonction ventriculaire gauche dans l'insuffisance aortique chronique asymptomatique. Etude par angioscintigraphie.

More accurate information is needed on the usefulness of radionuclide angiography performed during exercise for the assessment of left ventricular function in chronic aortic regurgitation and on its value compared with echocardiography. Between January, 1985 and January, 1988, we studied 23 asymptomatic patients presenting with severe, isolated and pure aortic regurgitation. Nine patients who were not operated upon during that period (group N) had the following characteristics: age 39.4 +/- 12.3 years, left ventricular end-diastolic diameter 67.3 +/- 4.7 mm, left ventricular end-systolic diameter 43.4 +/- 3.2 mm, left ventricular fibre shortening fraction 0.36 +/- 0.05, left ventricular radionuclide ejection fraction 0.67 +/- 0.10 at rest and 0.66 +/- 0.09 during maximum exercise. Compared with the values obtained in 8 controls of the same age (ejection fraction 0.65 +/- 0.07, p less than 0.05, at rest and 0.76 +/- 0.09, p less than 0.05, during maximum exercise), the behaviour of group N patients during exercise was perturbed. Fourteen patients who underwent surgery presented with the following characteristics: age 53.3 +/- 13.3 years (p less than 0.05), left ventricular end-diastolic diameter 71.4 +/- 8.7 mm (p less than 0.05), left ventricular end-systolic diameter 49.4 +/- 6.5 mm (p less than 0.05), fibre shortening fraction 0.31 +/- 0.03 (p less than 0.01), ejection fraction 0.53 +/- 0.08 at rest (p less than 0.001) and 0.40 +/- 0.08 during maximum exercise (p less than 0.001). These results suggest that radionuclide angiography performed during exercise is effective in the early detection and accurate evaluation of myocardial dysfunction in patients with chronic aortic regurgitation at the asymptomatic stage.(ABSTRACT TRUNCATED AT 250 WORDS)

[Radioimmunofixation: a new technic for characterizing immunoglobulins in unconcentrated cerebrospinal fluid. Preliminary results]. / Radio-immunofixation: une nouvelle technique de caractérisation des immunoglobulines dans le liquide céphalo-rachidien non concentré. Résultats préliminaires.

The restriction of heterogenicity of immunoglobulins G (IgG), or oligoclonal distribution, in the cerebrospinal fluid (CSF) can be observed after electrophoresis or electrofocalization, the IgG nature of each oligoclonal band being confirmed by subsequent immuno-electrophoresis. Using immunofixation, the oligoclonal bands can be visualized and characterized simultaneously, but requires previous concentration of the CSF, which is a source of error. The technique of radioimmunofixation, here described, allows the study of IgG with 20 microliters of non-concentrated CSF. It demonstrates that the oligoclonal characteristic is present in normal CSF and is a more quantitative than qualitative feature. Preliminary results show that the method can be applied to the study of all CSF proteins. By using a viral antigen labelled with I-125, for example, it should be possible to reveal and visualize the antibody activity of each oligoclonal band and to determine whether the quantitative increase in an oligoclonal band corresponds to a definite antigenic stimulation.

Comparison of six serum ferritin immunoassays and isoferritin spectrotypes in malignancies.

Serum ferritin was measured by six enzyme immunoassays in specimens from patients with digestive cancers (n = 30) and hematologic malignancies (n = 33). Most mean comparisons show significant differences in both groups of patients. In digestive cancers correlations between any two methods are very satisfactory (r > 0.99) but a proportional bias is often observed. In hematologic malignancies, correlations are bad (r < 0.80 in 8 out of 15 correlations) because of many discrepant values. Isoelectric focusing separation of isoferritins was performed in most specimens and the pattern of each serum was compared to the between kit CV. We conclude that an 'acid' spectrotype increases between-kit analytical variability. We try to explain the results taking into account the nature of the immunological systems and the cross-reactions with tissular isoferritins. In conclusion, our results indicate that large differences may be observed in sera from hematologic malignancies (leukemias, lymphomas ... ) We recommend that monitoring be achieved by the same method of measurement.

[Enalapril versus digoxin in chronic congestive cardiac failure. Study of variation of the left ventricular ejection fraction determined by radio-isotopic angioscintigraphy at rest and during exertion]. / Enalapril versus digoxine dans l'insuffisance cardiaque chronique congestive. Etude de la variation de la fraction d'éjection ventriculaire gauche déterminée par angioscintigraphie radio-isotopique au repos et à l'effort.

The effects of enalapril were evaluated in a double-blind, controlled study during 8 weeks, in patients with stable, congestive chronic cardiac insufficiency (functional classification II and III of the NYHA), in sinus rhythm, treated with digitalis and diuretics. 20 patients were randomly divided in two groups: one group continuing the digoxin (group A: 10 patients) and one group where enalapril was substituted for digoxin (group B: 10 patients). Patients from group B presented less clinical aggravation during the study. The left ventricular stroke volume (SV) is significantly decreased in group B at rest (0.21 +/- 0.06 at 50; 0.18 +/- 0.04 at 54; p less than 0.05), while it remained stable during stress. No variations of the SVs were noted at rest and during stress in group A. Considering its favorable clinical effects and after evaluation of its longterm side effects, enalapril may be an acceptable alternative to digitalis in cardiac insufficiency with sinus rhythm, except in patients for whom a drop in the systemic blood pressure or an increased kaliemia or creatininemia, could be potentially harmful.

[Colloid substitutes and hematocrit measurement by micromethods]. / Substituts colloïdaux et mesure de l'hématocrite par microméthodes.

A study was carried out to determine whether centrifugation and resistivity were reliable methods for measuring haematocrit of blood diluted with dextran or gelatin. The values obtained with the minicentrifuge Compur M 1100 (Bayer) (group 2) and with the Stat-Crit (Fumouze) (group 3) were compared with those obtained with the reference method (Coulter counter Model S + 2, Coultronics) (group 1). The study included 10 healthy subjects, aged 21 to 43 years. In each, sodium, potassium, chloride and protein concentrations were determined. Two further 10 ml blood samples were taken to be diluted with increasing amounts of either a fluid modified gelatin (Plasmion) or a dextran 40 (Piasmacair). The dilutions carried out were 0, 10, 20, 30, 40 and 50%. For each one, the haematocrit was measured using all three methods. The values obtained with undiluted blood were similar with the three methods. However, when blood was diluted with a dextran, haematocrit values were underestimated, whatever method was used, but by no more than 5% (difference not statistically significant). The result was the same with the centrifugation method for blood diluted with gelatin. On the other hand, values obtained with the resistivity technique for these samples were underestimated up to 15.5% (50% dilution; p less than 0.05). This was due to the negative electric charge of the gelatin, which lowered the haematocrit value in proportion to the concentration of gelatin. Although the technique is easier than centrifugation, measuring haematocrit with a resistivity method should be avoided in patients receiving a fluid modified gelatin.

[Fibrotic interstitial pneumopathies. Collagenolytic activity of the alveolar fluid]. / Pneumopathies interstitielles fibrosantes. Activité collagénolytique du liquide alvéolaire.

Collagenolytic enzyme release in alveolar structures is probably one of the initial events leading to impaired balance between collagen synthesis and degradation in the connective matrix of the lung, resulting in pulmonary fibrosis. The collagenolytic activity was determined in the bronchoalveolar fluid of 40 normal subjects or patients with miscellaneous pulmonary diseases and was found to be present in seventeen, viz.: 7/7 patients with interstitial fibrosis, irrespective of its origin: 4/4 patients with radiation pneumonitis; 4/15 patients with sarcoidosis and 2/2 patients with transient eosinophilic pneumopathy. There was no evidence of fibrosis in the 23 patients who showed no collagenolytic activity. Thus, collagenolytic enzymes are present in the alveolar structures of patients with interstitial pulmonary diseases of diverse origin capable of leading to fibrosis. Monitoring the release of this enzyme by bronchoalveolar lavage could be useful to evaluate the risk of fibrosis in such patients.

[Osteocalcin and smoking]. / Ostéocalcine et tabagisme.

Smoking is a risk factor for osteoporosis. Nicotine and nonnicotine tobacco smoke components have been shown to depress osteoblast activity in a number of in vitro and animal studies. To determine whether smoking is associated with depressed osteoblast activity in humans, we measured serum osteocalcin levels (using a radioimmunological method based on an antibody to human osteocalcin) in 24 male or female smokers and 24 matched nonsmokers. Overall, osteocalcin levels were significantly lower in smokers (15 +/- 6.95 ng/ml) than in nonsmokers (21.27 +/- 8.34 ng/ml) (p = 0.007). The difference between smokers and nonsmokers was significant in males (15.3 +/- 4.5 vs 23.27 +/- 9.7; p = 0.02) but not in females (16.27 +/- 8.9 vs 19.45 +/- 6.7; p = 0.2). These data suggest that smoking may induce osteoblast depression, either directly or via hormonal changes.