[The classical or laparoscopic operation in inguinal hernias]. / Operatie clasica sau laparoscopica în herniile inghinale.

AIM: Results evaluation in two trials of patients undergoing classical or laparoscopic surgery for inguinal hernia. MATERIAL AND METHOD: We compared 2 homogeneous trials of 80 patients with inguinal hernias treated by classic procedures: Bassini, Shouldice, Lichtenstein (trial I) or by laparoscopic approach with Plastex, Mercilene or Prolene prosthesis (trial II) between 1995-1997. RESULTS: Postoperative morbidity consisted in trial I in 5 seromas, 2 hematomas, 4 cases with neuralgic pain, 1 with testicular hypotrophy and 4 recurrences. In this trial the mean operative time was 22 min. and the mean hospitalization was 7 days. In trial II we registered a parietal bleeding at a lateral port imposing the conversion, 3 serohematomas, 2 recurrences by displacement of the prosthesis and 2 cases of neurologic pain. The mean operative time was 50 minutes and the mean hospitalization was 3 days. CONCLUSION: In spite of the longer operative time and the higher cost (the price of the prosthesis), in trial II the benefits of shorter hospitalization, lower morbidity and rapid socioprofessional reintegration are significant.

Growth hormone (GH) release by arginin-vasotocin (AVT) given intranasally in sexually immature children.

Eighteen healthy children were given 50 micrograms synthetic AVT intranasally and samples of serum for GH, magnesium and alpha-amino-nitrogen were drawn 30, 60 and 120 minutes later. There was a significantly different mean of GH peak level (13.1 +/- 3.02 microU/ml) which occurred generally at 30 minutes. The number of GH responders (i.e. peak level greater than 15 microU/ml) was of 7/18. The means of chemical determinations elicited little though opposite changes: slight increases in magnesium and slight decrease in alpha-amino-nitrogen. In a separate day, a sleep test of 30 minutes-duration performed in the morning resulted in a significantly greater GH peak level (28.2 +/- 9.7 microU/ml) but the number of GH responders was nearly the same (6/18). The serum magnesium levels following sleep decreased slightly and unsignificantly. The involvement of serotonergic neurons in mediating GH-releasing effects of AVT is briefly discussed.

Study on the innocuity of the hypophyseal human growth hormone.

The hypophyseal human growth hormone (hGH), a Raben type laboratory preparation, was re-evaluated as regards its innocuity for therapeutic use. Besides the usual control tests recommended by the Romanian Pharmacopoeia, the contamination of the hGH for clinical use with acute and slow viruses, was investigated taking into account the withdrawal of this hormone in many developed countries. The contamination was absent both with acute viruses as resulted from hemadsorption on cell cultures and counterimmunoelectrophoresis, and with slow viruses as observed from a two year-follow up of guinea pigs injected intracerebrally with the hGH preparation. Further, the content of the growth hormone itself as well as the contamination degree with other pituitary hormones was examined. The hGH-RIA content was 2.23 +/- 0.13 IU/mg (means +/- SEM), range: 1.38-2.80 IU/mg (1st International Standard hGH 80/505-1982). The prolactin contamination assessed by RIA was 187.34 +/- 37.66 ng/mg hGH, range: 28.44-385.20 ng (International Standard WHO: 80/541). The LH and FSH contamination as quantified by the Isocommerz (DDR) RIA kits was with two orders of magnitude lower than 10 IU-LH/IU-hGH, the upper LH contamination limit considered as acceptable. Moreover, the proportion of the large molecular forms in the lyophilized hGH preparation was investigated by polyacrylamide gel electrophoresis. Corroborating the data obtained by these control tests with our previous experience on 149 pituitary dwarfs treated with this hGH preparation during the interval 1964-1984, resulted minor risks of some dangerous side effects of hGH administration in children with growth hormone deficiency by possible contamination with pathogenic agents or with other disturbing hormones.

Thyroid hormone-induced reduction of urinary 5-hydroxyindole acetic acid (5 HIAA) in obese children. Comparison with hypothyroid patients of similar age having either pituitary dwarfism or congenital myxedema.

Urinary excretion of monoamine metabolites (noradrenaline-NA, adrenaline-A, 3-methoxy-4-hydroxyphenyl glycol-MHPG, homovanillic acid-HVA, 5-hydroxyindole acetic acid-5 HIAA) was studied in four groups of children as follows: Group I consisting of obese children subjected to caloric restriction and to a short term course of thyroid extract in "low" dosage (1-2 mg/kg bwt), Group II consisting of obese children subjected to diet alone, Group III consisting of children myxedema and subjected to a short term course of thyroid extract given in the "high" dosage (3-5 mg/kg bwt) and Group IV consisting of GH deficient short children having (many of them) thyrotropin deficiency and subjected to a short term course of thyroid extract in "very high" dosage (5-10 mg/bwt). In obese, calorie-restricted children, the previously low mean level of 5 HIAA excretion was further lowered by thyroid extract. In obese children subjected to calorie restriction alone no urinary abnormality was noted. The congenitally hypothyroid patients had low levels of basal 5 HIAA when compared to controls. The degrees of 5-hydroxy tryptamine (5 HT) deficiency in Group III was similar to the obese groups. The thyroid extract course did not influence, at least in short term administration, the low 5 HIAA levels in group III. In GH deficient, short children (group IV) thyroid extract had no significant effect on urinary pattern of monoamine metabolites. A central 5 HT deficiency may tentatively explain the mood disturbances and possibly the other psychic disorders in both the obese and myxedematous patients. The different effects of thyroid extract on 5 HIAA may also witness the differences in the food intake behaviour in these two conditions.

Growth deficiency with high circulating growth hormone levels: the insulin stimulation test in 39 children.

Thirty nine patients with abnormal high basal hGH levels were selected and analysed as a part of a retrospective study of the results of 1,500 insulin stimulation tests (IST), applied in children and adolescents with growth deficiency. Their height, weight, and bone age were lower than their corresponding chronological age. Both in girls and in boys groups, responders and nonresponders subgroups were detected as judging by the results of the secretagogue action of insulin on hGH. The hGH basal levels were 43.88 +/- 18.27 microU/ml (X +/- SD) in boys (no = 22) and 56.61 +/- 35.21 microU/ml in girls (no = 17). It is to be noted that the hGH nonresponder group had deeper hypoglycemia at 30 minutes post-insulin injection than the responder group: 53.6 +/- 13.0 mg/100 ml (X +/- SD) vs 66.0 +/- 11.5 mg/100 ml respectively (p less than 0.01). Two siblings, a girl and a boy, had the highest basal and stimulated hGH, either during the IST or starvation. One of them, the boy, during the starvation test, had a paradoxical fall of about two orders of magnitude of the serum hGH 4 hr after basal sample collection. These two siblings are similar to the familial Laron type dwarfism. The possible mechanisms of growth deficiency in children with constant high but variable hGH values are discussed, as well as the aspects concerning the therapeutic ways to improve their linear growth.

The serum osteocalcin levels in adult and aged hypothyroid patients.

Osteocalcin (OC) or the bone protein containing gamma-carboxyglutamic acid (BGP or Gla-P), is a specific and sensitive marker of bone turnover. A radioimmunoassay (RIA) system for human osteocalcin was developed with the sensitivity of 0.5 ng/ml. The osteocalcin was measured in sera from 33 hormonally and/or clinically hypothyroid patients: 12 adult and 21 aged patients. For comparison, blood samples were collected from 14 hormonally hyperthyroid adult patients in whom the OC levels were 16.23 +/- 7.57 ng/ml (mean +/- SD) and from hormonally euthyroid adult patients (previously treated hyperthyroid patients) having OC 9.76 +/- 5.32 ng/ml. Abnormal low OC levels were noted in the hypothyroid adult patients group: 1.04 +/- 0.23 ng/ml by comparison to the hypothyroid aged patients 3.76 +/- 2.38 ng/ml (p less than 0.001). Moreover, great variability of the OC serum levels was observed in the aged group, four patients hormonally eu- or hypothyroid having high OC levels in the range: 13.29-55.45 ng/ml and other three patients although hormonally euthyroid but clinically hypothyroid had low OC levels 0.88-2.27 ng/ml. The abnormalities of the OC levels in hypothyroid adult and aged patients are discussed.

Methods of isolation and purification of bovine and rat osteocalcin.

Measurement of osteocalcin (a bone protein containing gamma-carboxyglutamic acid, GLA-P) in the biological liquids and tissues is of interest for studying the mechanisms of bone diseases. It is also helpful in making a diagnosis and in following up the patients with diseases in which the osseous system is affected. Methods have been worked out for isolation and purification of osteocalcin from bovine and rat femur in order to use the purified substance as a starting material for developing radioimmunoassay (RIA) systems for each of these osteocalcins. The work went through the following stages: the processing of the femur bone, preparation of the 200 microns bone powder, demineralization of the powder, and concentration of the bone extract, purification of the extract by molecular exclusion chromatography on Sephadex G-100 and repurification to the product by ion exchanger chromatography in NaCl linear gradient, on DEAE Sephadex A-25. The purity of the substance obtained was tested by disk electrophoresis in polyacrylamide gel. The bovine and rat osteocalcins obtained had the adequate purity for being used as RIA reagents.

The serum osteocalcin levels in patients with thyroid diseases.

The mechanism of bone, calcium, phosphorus and proteins abnormalities observed in hyperthyroidism is rather complex and as yet not wholly understood. Increased serum osteocalcin was recently reported in hyperthyroid patients and its decrease after 4-8 months of treatment. Osteocalcin was measured by RIA in the sera of 211 women and 18 men with thyroid diseases. The patients were divided into 3 groups according to diagnosis: I. polynodular goitre and subacute thyroiditis (59 women, 5 men); II. Graves' disease (70 women, 3 men) and III. thyroid cancer, after treatment by surgery and 131I (82 women, 10 men). The osteocalcin levels in the sera of these patients were: 2.97 +/- 2.63 ng/ml (mean +/- SD) for the women and 3.56 +/- 2.10 ng/ml for the men in the 1st group; 16.31 +/- 11.34 ng/ml for the women and 12.75 +/- 6.09 ng/ml for the man in the IInd group and, 1.01 +/- 0.60 ng/ml for the women and 0.78 +/- 0.46 ng/ml for the men in the IIIrd group. No differences were found between the osteocalcin concentrations in the hyperthyroid female patients treated with antithyroid drugs (no = 58) and the non-treated hyperthyroid women (no = 12): 16.22 +/- 11.40 ng/ml vs 16.74 +/- +/- 11.53 ng/ml. These data suggest that bone resorption stimulated by endogenous thyroid-hormones is a rather resistant processus, persisting even after 6-8 mos of associated anti-thyroid therapy. Further are analyzed the possible causes of the subnormal osteocalcin levels observed in patients with thyroid cancer treated by surgery and radioisotope, whose suppression therapy was discontinued 2-3 weeks before blood sampling.(ABSTRACT TRUNCATED AT 250 WORDS)

Sleep-induced GH release in otherwise GH deficient children having short stature; a possibly new variant of GH neurosecretory dysfunction (GHND).

Sleep and exercise are considered physiologic stimuli of growth hormone (GH) release from the pituitary. They were used to some extent for the assessment of GH reserve in growth disorders of supposedly pituitary origin. In order to better establish the diagnostic value of the sleep test, we studied 34 children of both sexes aged 6-14 having various degrees of shortness of stature associated with bone age retardation (range of the latter: 3-10 years). Eight healthy controls of the same age were also subjected to the sleep test. The latter was begun at 10:00-11:00 a.m. generally in a dark room under polygraphic control. The subjects were fed a standard breakfast at 8:00-8:30 a.m. After thirty minutes of steep or after reaching the IVth stage of slow-wave sleep, the children were awakened and samples of venous blood for GH determinations were drawn. Two other GH reserve tests were also performed in other days, usually insulin-induced hypoglycemia and glycine intravenous loading. Except sleep, no other GH provocative tests were performed in controls. In normal GH reserve children (21/34) according to other tests, the sleep test was positive (i.e. peak values greater than 15 microU/ml) in 7 cases and the mean GH response was not significantly smaller than the peak recorded after insulin-induced hypoglycemia. In abnormal GH reserve, presumably hypopituitary children (13/34) according to other tests, the sleep confirmed the other tests in 8 cases. In five cases of clinically and endocrinologically "hypopituitary" dwarfism, the sleep test revealed a normal response.(ABSTRACT TRUNCATED AT 250 WORDS)

Radioreceptor assay of human prolactin using rabbit mammary receptors.

The purpose of this work is to develop the procedures for the preparation of the reagents suitable for the radioreceptor assay (RRA) of human prolactin (Prl). Human purified Prl (NIAMDD-hPrl-16) was labelled with 125I by the Chloramine-T or alternatively by the lactoperoxidase method. As reference preparation we used Prl isolated from the ethanolic step of the routine procedure for the preparation of human growth hormone (hGH) for clinical purposes. The lactogenic receptors were prepared from the pregnant rabbit mammary gland previously stimulated with insulin, cortisone and dried thyroid extract. The final receptor preparations obtained by ultracentrifugation contained 8.85-39.36 mg protein per ml. The prolactin was measured in the human sera and in our hPrl preparations by a double antibody radioimmunoassay (RIA) system using the NIAMDD reagents. We developed a RRA system for hPrl using rabbit mammary receptor preparations with a protein concentration of about 4 mg/ml. The comparative competition showed the same magnitude of the inhibition of the tracer receptor binding of hPrl and hGH. This interference of hGH makes difficult the assessment of the specificity of the hPrl-RRA system otherwise accountable by the structural and/or biological relationship of the three lactogenic hormones: hPrl, hGH and human placental lactogen (hPL). Studies concerning the preparation of a purified and solubilized rabbit mammary receptor and of an antiserum for it are in progress in our laboratory with the objective to provide a useful tool for the investigation of the lactogenic receptor structure and function relationship.

Serum-to-saliva transfer of the immunoreactive insulin (IRI) in children with obesity associated with insulin-resistance.

Assuming that the serum-to-saliva transfer of insulin reflects internalization and re-cycling of the hormone in the membrane-located binding sites of salivary epithelial cells and that these cells have in obesity a'marked decrease in insulin receptor content, it has been postulated that insulin resistance in infantile obesity can be detected by the changes in the salivary immunoreactive insulin during the oral glucose tolerance test (OGTT). The study included 31 obese children and adolescents of both sexes, subjected to OGTT. Samples of blood and saliva were collected at 30, 60, 120, 180 and 240 minutes for determinations of glucose and IRI. The blood glucose values were generally normal whereas IRI was excessively high. The dynamics of salivary IRI was similar (easy peak followed by slow descent) with the mean serum values but lower by about two-thirds, and the peak was 30-60 minutes delayed. The serum IRI values correlated significantly with the saliva ones at all time-intervals except for the 30-minute ones. The serum IRI values were significantly lower at the 30-minute time interval, whereas the salivary IRI were the lowest (and of borderline significance) at the 60-min. time interval. The mean glucose/kg doses given orally were not significantly different in the two groups. It was concluded that a hormonal activity detectable by IRI assay through the PEG separation method does exist, with a concomitant variation of serum-to-saliva transfer as shown by the OGTT test. It was also concluded that since the salivary values are lower, the direction of the flow is from serum to saliva and not the reverse. Finally, on the basis of our data, an "in situ" synthesis of insulin (hormonogenic exocrinism) can not be ruled out.

A heterologous radioimmunoassay system for human osteocalcin. Applications in performance sports girls.

Taking into account the involvement of the measurement of osteocalcin, a protein-containing gamma-carboxyglutamic acid (Gla), in investigating the physiopathology of bone and calcium metabolism, a radioimmunoassay (RIA) method useful in human clinics was developed. The reagents for the RIA of human osteocalcin namely, the radioiodination preparation, the reference preparation and the antiserum were obtained starting from bovine bone on account of the structure similarities between bovine and human osteocalcin. The reagents were incubated at 4 degrees C for 48 h followed by the polyethylene-glycol (PEG) separation technique. The sensitivity of the osteocalcin RIA method is 2.0 ng/ml, the intraassay precision is 3-5% and the interassay precision, 8-12% (coeficient of variation). Osteocalcin was measured in the sera of 3 groups of performance sports girls under basal conditions and being of more or less similar ages: rowing girls (n = 10, mean age 16 yr 6 mos), gymnast girls (n = 13, mean age 14 yr 8 mos) and table tennis girls (n = 9, mean age 16 yr). The same samples were used for the measurement of myoglobin. Osteocalcin quantification showed similar levels in the rowing and the table tennis girls, i.e. 11.91 +/- 3.39 ng/ml (X +/- SD) and 9.50 +/- 4.15 ng/ml, respectively. By contrast, in the gymnast girls, the osteocalcin levels were about 4 times higher (44.31 +/- 7.27 ng/ml) than in the other two groups. Myoglobin measurements showed lower levels in the gymnast group, than in the rowing and the table tennis groups, the latter two having similar levels. The multiplicative increase of osteocalcin in the gymnast girls is interpreted and commented in relation to their age and sexual development.

The measurement of the peptide hormones in saliva. 2. Chorionic gonadotropin subunits in pregnancy.

The alpha and beta subunits of the human chorionic gonadotropin (HCG) were measured during pregnancy in the saliva of a first group (no. = 23) and in the serum and saliva of a second group (no. = 25). Only ten pregnant women had normal gestation, the remaining 39 being diagnosed as miscellaneous pathologies, the threatened (or impending) abortion being the most frequent (no. = 19). The mean serum levels of alpha-HCG increase from 75.97 +/- 18.59 ng/ml (X +/- SEM) during the first trimester to 341.98 +/- 65.09 ng/ml during the third trimester of the gestation. The mean salivary concentration of the alpha HCG seems unmodified during pregnancy (approximately 10 ng/ml), but large interindividual variations were observed (limits 0.10-28.78 ng/ml) possibly due to the non-homogeneity of the investigated groups. The presence of the beta-HCG subunit in saliva could not be assessed, the great part of the values being aggregated around the sensitivity limit of the RIA technique (0.2 ng/ml). The physiological significance of the presence of the HCG and of its subunits in saliva as well as the ways to elucidate the possible selective role of the blood-saliva barrier are discussed.

A radioimmunoassay system for LH based on the antigenic similitude of LH with HCG: methodology and applications.

A double antibody radioimmunoassay (RIA) system for LH (LH-IEP Kit) was developed using the antigenic similitude of LH with HCG. The first antibody (Ist Ab) is rabbit anti-HCG serum, initial dilution 1:200 000. The tracer is 125I-HCG (code MJ-14 Swierk Poland). The standard curve is calibrated with the reference preparation hLH-Ist-IRP 68/40 kindly offered by WHO. The IInd Ab is pig anti-rabbit IgG serum. The incubation conditions: volume-0.3 ml; time-24 hrs with Ist Ab and 24 hrs with the IInd Ab at ambient temperature. The sensitivity of the RIA system for LH is 1.5 mIU/ml. To validate our RIA system the LH was measured in the serum samples collected from 9 women during the menstrual cycles, from 2 boys during the GnRH test, from 2 amenorrheic women and from 20 children, adolescents and adults with miscellaneous pathologies. In all these samples, parallel measurements of LH and FSH were performed using DDR commercial RIA Kits-SSW. It is to be mentioned that the LH-RIA Kit-SSW is not completely homologous, the Ist Ab being rabbit serum anti-HCG. The results obtained during 4 menstrual cycles, in which the LH peak is observed around the mid point of the interval are: follicular phase 17.92 +/- 5.58 mIU/ml (means +/- SD) with LH-IEP-Kit and 5.51 +/- 2.77 mIU/ml with the LH-SSW-Kit, peak: 26.07 +/- 22.13 mIU/ml and 9.13 +/- 5.60 mIU/ml respectively; luteal phase: 12.55 +/- 5.46 mIU/ml and 3.4 +/- 2.38 mIU/ml, respectively. The LH values observed by the two kits through all 9 menstrual cycles are well correlated ("r" values in the range 0.7-0.9) but high discrepancies were observed in the remaining 3 cycles ("r" between 0.07 and 0.6). These discrepancies as well as those observed in some adolescents with genetic anomalies and in a patient at climacterium are suggesting that the two LH-RIA systems measure not only a common molecular area but also different areas of the LH circulating molecules.

Radioreceptorassay of insulin using human erythrocytes. I. Methodological aspects.

It is known that human erythrocytes have specific receptors for insulin. This works describes a radioreceptor assay (RRA) suitable for the determination of the insulin binding to the surface receptors of erythrocytes. The erythrocytes were isolated from heparinated blood collected in the morning after overnight fasting. In the incubation mixture are 4-5 X 10(9) cells/ml, labelled porcine insulin and nonlabeled (cold) insulin in serial concentrations, range 10-10(5) ng/ml. Incubation (150 min. at +15 degrees C) is ended by suspending the incubation mixture in cold buffer. After washing, the tracer insulin bound to erythrocytes is measured on a gamma counter and the percentage of the specific binding is calculated against the total radioactivity determined prior to the washing steps. By this procedure, the maximal binding of 125I-Insulin was 10.55 +/- 0.78% (mean +/- SD) in five normal volunteers and 6.79 +/- 1.77 (mean +/- SD) in five obese nondiabetic patients, having an enhanced insulin response in the oral glucose tolerance test. The slight hemolysis that happened accidentally in some tubes after incubation caused rude degradation of the tracer, the binding being decreased at last by 25%. The method described (according to Gambhir K. K. et al 1977 slightly modified) is specific and thus suitable for estimating the prognosis of obese children, patients with acute and chronic hepatitis and also for the study of the role of erythrocytes in insulin metabolism and possibly in the metabolism of other hormones.

A radioimmunoassay system for human myoglobin: method development and applications.

double antibody radioimmunoassay (RIA) system for human myoglobin (hMb) was developed using our own reagents. The antigen (hMb) was isolated from human muscle, purified and stored frozen until needed for immunization, radiolabeling or reference preparation. The anti-hMb serum raised in rabbits was used at 1:2.10(4) dilution (initial). The Chloramine-T method was used for the hMb labeling obtaining at 10-15 muCi/micrograms (370-550 KBq/micrograms) specific activity. Working standards were prepared having concentrations in the range of 2.0 to 500 ng/ml. The reagents were incubated at +4 degrees C for 48 plus 24 hrs. The specificity and accuracy of our hMb-RIA system were validated using for parallel assays an already validated immunochemical system, the hemagglutination inhibition (HI) technique and the parallelism test using serum dilutions from patients with acute myocardial infarction (AMI). The serum hMb concentration in normal subjects (no = 23) was 54.14 +/- 15.08 ng/ml (X +/- SD), being higher in short-term hypothyroidism (no = 13), 87.95 +/- 20.90 ng/ml (p less than 0.0005) or in treated hyperthyroidism (no = 5), 80.03 +/- 21.81 ng/ml. In AMI (no = 6) the serum hMb concentration varied in the range of 123 to 1510 ng/ml. The sensitivity of our hMb-RIA system is 2 ng/ml and the intraassay average error (coefficient of variability % in %B) is 2.26%. Trials to shorten the incubation time showed that adequate binding of labelled Mb may be obtained with 2 plus 4 hr intervals at room temperature. It is necessary to establish, in our conditions, the variation limits for serum hMb in normal subjects according to sex and age as a comparison basis for the study of its physiological and pathological variations.

The simultaneous isolation of the human pituitary hormones. II. The luteinizing hormone.

The main purpose of the work is the isolation of luteinizing hormone (LH) from frozen human pituitaries. The procedure for the processing of the glands included the steps described by P. Roos [22] for the preparation of the follicle stimulating hormone (FSH) slightly modified and adapted to our conditions. After thawing, the pituitaries were homogenized in 0.03 N Naphosphate buffer, pH 5.7. The protein bulk containing growth hormone (GH) was separated by precipitation with saturated ammonium sulfate (v/v). The supernatant containing gonadotrophins, obtained by centrifugation, was fractionated by chromatography on DEAE cellulose, the first peak being further fractionated by column gel-filtration on Sephadex G-100 after a previous concentration by dialysis against polyvinylpyrolidone 10%. The first peak obtained by this last fractionation was considered for the time being, as pure LH. The extraction and fractionation steps were followed by disc polyacrylamide gel electrophoresis and by radioimmunoassay measurement of the LH concentration. Studies concerning the homogeneity and the biological activity of our LH preparation by comparison to the LH preparations recommended by WHO, are in progress in our laboratory.

An improved procedure for the preparation of the human thyroglobulin and the development of a thyroglobulin autoantibody kit.

Human thyroglobulin has various applications as a diagnosis reagent or for studying the physiopathology of the protein and hormonal biosynthesis in the thyroid gland. The previous preparation procedure for isolating the human thyroglobulin currently used in our laboratory, has some inconveniences as regards the low yield and the noxious influence of the ammonium sulfate precipitation upon its molecular integrity. Therefore, we describe an improved fractionation and purification procedure whose main steps are extraction of the thyroid tissue homogenate in 0.15 M NaCl followed by a double gel filtration on Sephadex G-200 column. In this way are obtained thyroglobulins A and B grade using the two chromatographic steps, respectively. Thyroglobulin B grade is further submitted to preparative polyacrylamide gel electrophoresis for separating some thyroglobulin isomers as recognized by other groups using the analytical ultracentrifugation procedure. The different fractionation and purification steps were checked by double diffusion in gel using rabbit anti-thyroglobulin serum and horse antihuman serum protein. The homogeneity and the molecular weight of the different fractions we evidenced were analyzed by the aid of disc and plate electrophoresis in polyacrylamide gel. The authors developed the technique for thyroglobulin autoantibody detection by the passive haemaglutination method using stabilized erythrocytes coated with thyroglobulin A-grade. Thyroglobulin B-grade used as a tracer and a reference preparation in a RIA system offered a sensitivity of 1.5 micrograms/liter for thyroglobulin detection in biological fluids.

Studies of the reproduction function in hyper- and hypoadrenocorticism.

The investigation was carried out on 66 patients with hypo- or hyperfunctional syndromes of adrenocorticism, hospitalized and treated by our team. The patients were grouped into 5 lots according to their diseases. Lot one consisted of 34 female patients with Cushing's syndrome, lot two of 10 males with Cushing's syndrome, lot three of 10 males with Addison's disease and lot four, of 6 females with androgenic hypercorticism. The morphofunctional disorders of reproduction were followed up clinically and by complex hormone assays, before and after treatment. The incidence of these disorders is very high, and the results of clinical observations and of laboratory data demonstrate that both the excess of adrenocortical hormones and the decrease in their circulating level have a negative influence on the reproduction function or represent a risk factor in cases of pregnancy. By the curative treatment of the adrenal cortex a preventive treatment of abortion and/or sterility is achieved.