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The adenosine receptors in the plasma membrane of chromaffin cells from bovine adrenal medulla were characterized. The presence of A1 receptors was discounted owing to the absence of R-[3H]phenylisopropyladenosine (R-PIA) and [3H]8-cyclopentyl-1,3-dipropylxanthine ([3H]-DPCPX) binding. The binding of the specific A2a ligand CGS-21680 was low. In contrast, the binding of 5'-(N-[3H]-ethylcarboxamidoadenosine ([3H]NECA) was relatively high (1.7 pmol/mg of protein at a ligand concentration up to 90 nM). This binding did not correspond to non-adenosine receptor NECA binding sites because the specific [3H]-NECA binding was similar when unlabeled adenosine, NECA, or R-PIA was used to measure the nonspecific binding. The rank order of potency of different ligands for the displacement of specific [3H]NECA binding was DPCPX greater than NECA greater than chloroadenosine greater than R-PIA greater than theophylline = CGS-21680. These results indicate that the receptors present on the plasma membrane of chromaffin cells are exclusively of the A2b subtype.

Administration of Benzo(a)pyrene (BP, 50 mg/kg/d) to pregnant rats significantly increased Glutathione S-transferase (GST) activity in placental tissue-extract (Vmax = 40 nmol/min/mg protein and 69 nmol/min/mg protein in controls versus treated animals respectively; P less than 0.01) and total fetal tissue-extract (Vmax = 51 nmol/min/mg protein and 82 nmol/min/mg protein in controls versus treated animals respectively; P less than 0.01) indicating an induction effect of BP on the GST system. An increase in the Km values was also observed: 1.61 x 10(-3) M and 2.84 x 10(-3) M in control versus treated placentae; 1.38 x 10(-3) M and 2.05 x 10(-3) M in control versus treated fetuses. A competitive effect on the enzyme by the BP present in the sample may also be involved. The glutathione content in both tissues did not show any changes after the treatment with BP. This increase in the GST system was not sufficient to protect the fetus. BP affected the reproductive performance of pregnant rats by significantly increasing the number of resorptions and fetal wastage, and, also, by decreasing the fetal weight.

We present our experience of 44 cases with unilateral vesicorenal reflux that had been treated by ureteroneocystostomy using the Cohen technique. The reflux disappeared in 32 cases, it appeared in the contralateral ureter in 7 patients, it was ipsilateral in 2 and bilateral in 3. Of the 10 cases with contralateral reflux, 2 required surgery, while 8 were amenable to medical treatment. In our series, contralateral reflux appeared at random. Since the spontaneous cure rate was 80%, we believe that bilateral antireflux surgery for unilateral reflux is unwarranted. Urodynamic studies can be important in the indication for conservative treatment.

Pig brain cerebral cortex was subfractionated by isopycnic centrifugation in sucrose gradients. In each subfraction the content of the agonist [3H]R-PIA binding, the activity of adenosine metabolizing enzymes (5'-nucleotidase and adenosine deaminase) and the activity of membrane marker enzymes were determined. The fractions were also examined by electron microscope. In general, the results suggest a widespread distribution of A1 adenosine receptors in membranes from different origins. Marker enzyme profile characterization indicated an enrichment of A1 adenosine receptor in pre-synaptic membranes isolated from the crude synaptosomal fraction (P2B subfraction) as well as in membranes of glial origin such as myelin. The receptor is also present in the endoplasmic reticulum and in membranes isolated from the microsomal fraction that seem to have a post-synaptic origin (P3B). In subfractions having a high content of adenosine receptor the equilibrium binding parameters were obtained as well as the proportion of high- to low-affinity sites. From the values of the equilibrium constants it was not possible to find differences between the receptor in the different subfractions. Analysis of the affinity state distribution showed a diminished percentage of high-affinity sites in fraction P3A, which can be accounted by the existence of myelin membranes; in contrast the percentage of high-affinity states was higher in P2 and P3B, indicating that in these fractions the receptor is present in synaptosomal membranes. The close correlation shown between the enzyme 5'-nucleotidase specific activity and the specific ligand binding distributions led us to postulate an important role for the enzyme in the regulation of adenosine action in pig brain cortex.

A method of obtaining estimates of the maximum binding and association and dissociation rate constants for a receptor-ligand interaction is described. This new procedure, the association-saturation method, is based on an exact mathematical equation which defines the model without simplifications. The method proposed is readily applicable to any system consisting of one ligand and one receptor. With only four determinations, each involving one ligand concentration and an association time, it is possible to determine the number of specific binding sites, the dissociation and association rate constants and the equilibrium dissociation constant with accuracy. No dissociation curve is required, and only one point of the association curve for each ligand concentration is necessary. In addition to the higher confidence in the estimates of the parameter values obtained, this method leads to important savings in radiolabelled compounds and experimental time. The results were compared with those obtained with standard association-dissociation curves and saturation isotherms. The optimum number of replicates of each experimental point to obtain reliable estimates of different parameters is discussed on the basis of simulation studies. The performance of the procedure is analysed by means of association-saturation experiments with the agonist [adenine-2,8-3H,ethyl-2-3H]N6-phenylisopropyladenosine and solubilized A1 adenosine receptors from pig brain cortex.

The binding of [3H]N6-(R)-phenylisopropyladenosine ([3H]R-PIA) to pig brain cortical membranes was investigated as a function of membrane fluidity and of the physicochemical characteristics of the medium. The two affinity states of the A1 adenosine receptor behave differently in the experiments performed. Increases (up to 6.7 poises) or reduction (up to 1.3 poises) in membrane microviscosity did not lead to significant variation of equilibrium parameter values except for a slight modification of the low-affinity Kd value at 1.3 poises. Addition of cholesterol to raise the microviscosity up to 9.0 poises led to a marked decrease of binding to the high-affinity state without modifying the Kd. Furthermore, the Kd value for the low-affinity state rose markedly. Addition of sucrose, which modifies the conductance, viscosity and density of the aqueous medium, did not lead to significant changes when used at a concentration of 0.25 M, either in isolation or in radioligand binding assay medium. The presence of 0.32 M sucrose in the isolation medium led to a 30% fall in the total binding without affecting the distribution and Kd values for either affinity state very much. However, the presence of greater than or equal to 0.32 M sucrose in the binding assay medium produced the disappearance of the low-affinity states and the appearance of high-affinity states; only one affinity state was found to have a somewhat increased Kd value. On the other hand, increases of the medium conductance did not lead to the disappearance of the low-affinity state although some decrease of the binding was observed at high Tris concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)

By immunocytochemical methods, using colloidal gold particles coupled to antibodies, it is possible to label the occupied nuclear T3 receptors and observe them by electron microscopy. We carried out some experiments to validate these methods by testing the absence of non-specific binding of the antibodies to subcellular structures different from the T3 receptors. So, we have verified that the number of occupied T3 receptors in tissues from normal rats is lower in kidney cells than in heart and liver ones, a finding which is an agreement with data reported by others. The occupation of the receptors increases after T3 administration and decreases by triiodothyroacetic acid (Triac) administration to the rats, due to the displacement of T3 from the receptors with the latter. Our results are similar to those of the other authors using radioactive tracers. Non-specific binding was not detected. The occupied T3 receptors labelled with colloidal gold particles are located most commonly in the hetero/euchromatin interface. From our results we conclude that the colloidal gold particles observed by electron microscopy specifically label occupied T3 receptors, and the determination of the thyroid status at cellular level can be achieved using electron microscopy after labelling the receptors by immunocytochemistry with colloidal gold particles coupled to antibodies.

The effect of sulphydryl reagents (N-ethylmaleimide-NEM-4- hydroxymercuriobenzoate-HMB- and 5-5'-dithio-bis-2-nitrobenzoate-DTNB-) on agonist and antagonist binding to A1 adenosine receptors from pig brain was studied. The action of the mercurial agent HMB was found to be strong and seemed to be nonspecific. The effects of either NEM or DTNB were milder and more specific. The characterization of the agonist binding in membranes pretreated with moderate concentrations of DTNB and NEM led to reduced affinities for both high- and low-affinity sites without marked modifications of maximal binding or of proportion of affinity states. These results for NEM are surprising since the compound is usually used to mimick the effects of Gpp(NH)p, i.e. to shift high-affinity states to low-affinity states. It was found that this Gpp(NH)p-like effect of NEM is only possible when the compound is included in the assay medium. Similarly, Gpp(NH)p produces the uncoupling of the receptor molecule from G protein if included in the assay medium. Thus, membranes pretreated with Gpp(NH)p exhibited both affinity states and with similar equilibrium binding parameter values to those of the crude membranes.

The effect of phospholipases and proteases on the membrane-bound and solubilized A1 adenosine receptor has been studied. Phospholipids modulate the [3H]N6-(R)-phenylisopropyladenosine binding to A1 adenosine receptors in crude membranes and in soluble preparations, because changes in the phospholipid environment decrease both the binding capacity and the affinity for the ligand. It has become clear that 1) there is co-solubilization of receptor and phospholipids; 2) the phospholipid requirements are different for the coupled and the uncoupled receptor; 3) a net charge in the polar head produced by phospholipase D prevents the agonist binding to the receptor-G protein complex; alternatively, when the whole polar head is removed by phospholipase C the uncoupled receptor is altered; and 4) the protease action upon the receptor suggests that receptor coupled to G protein is more protected by the membrane than the uncoupled receptor. In kinetic experiments performed on membranes it was demonstrated that phospholipase C and trypsin increased the Kd value of the high-affinity state by modifying both k1 and k-1. In contrast they only modified the dissociation constant of the low-affinity state. In conclusion it should be noted that phospholipids play a key role for the binding of R-PIA to A1 adenosine receptor. Also, a different disposition within the membrane of the coupled and uncoupled receptor is encountered.

In this article the existence of A1 adenosine receptors and the absence of A2 adenosine receptors in myelin membranes purified from pig brain white matter are demonstrated. The characterization of (R)-[3H]phenylisopropyladenosine ([3H]R-PIA) binding to purified myelin fractions was performed. The distribution of high- and low-affinity species of the A1 adenosine receptor was different in heavy, medium, and light myelin. The fluidity of myelin subfractions and of pig brain cortical membranes was estimated; the microviscosity of heavy myelin (5.4 poises) and of cortical membranes (5.1 poises) was similar and less than that of medium (7.8 poises) and light (8.2 poises) myelin. It was also demonstrated that the agonist R-PIA modifies the microviscosity of myelin membranes and that the degree of modification depends on the fluidity of the membrane assayed. These results suggest that adenosine receptors may have an important role in the functionality of myelin membranes.

Kinetic evidence for negative cooperativity on the binding of [3H]R-PIA to A1 adenosine receptors was obtained from dissociation experiments at different ligand concentrations and from the equilibrium isotherm. The dissociation curves indicate that there is an apparent ligand-induced transformation of high- to low-affinity states of the receptor. At concentrations of 18.2 nM R-PIA or higher there was only found the low-affinity state of the receptor. In view of these results equilibrium binding data were analyzed by the usual two-state model (assuming that there is an interconversion between them) and by the negative cooperativity model employing the Hill equation.

The polluted environment found in highly industrialized areas and in big cities contains a great quantity of electrophilic (EC) and proelectrophilic (PEC) compounds, which largely contribute to the development of several pathological processes in humans. EC and PEC can be coupled to the cysteine moiety of glutathione spontaneously or by the glutathione S-transferase system (GST), giving nontoxic metabolites that can be eliminated as urinary thioethers (UT). Therefore one approach to establishing the degree of impact of EC and PEC on the human body is the analysis of UT in the population living in polluted environments. The work presented here has been carried out in the city of Barcelona with a group of 50 children living in a polluted area, over a 12-mo period. Our results demonstrate that UT are significantly higher when the amounts of air pollutants (AP) increase; although the level of contamination never exceeded the established "safe limits," UT reached values similar to those found in people smoking more than 10 cigarettes/d. These results may contribute to establishing the maximal levels of contamination compatible with a healthy life, on the basis of patterns of true salubrity rather than on political and economic criteria.

Synopsis A prophylactic antistriae cream (Centella asiatica extract, alpha-tocopherol, and collagen-elastin hydrolisates) was assessed by a double blind trial in 80 pregnant women. In the placebo group 22 women (56%) presented striae, whereas in the treated group only 14 women (34%) developed striae in this pregnancy; this difference was significant (p < 0.05; chi(2) test). An arbitrary score was designed to assess the intensity of striae (from 0 to 3); this score was 1.42 (sd 0.5) in the treated group and 2.13 (sd 1.32) in the placebo group and this difference was also significant (p= 0.014; Mann-Whitney test). In women with a history of striae during puberty, the active cream induced a significant absolute prevention in 89% of the cases whereas in the placebo group all the women developed striae (p= 0.00014; chi(2) test).

Tuberculin test with PPD RT-23 with Tween 80 (2 TU strength), was performed to 228 infants under two years of age. None of them had any history of contact with tuberculosis. All were healthy, well-nourished infants, and had been vaccinated with BCG at birth. A positive PPD reaction (greater than 6 mm), was found only in 8.8%, of them and 16.2% had no BCG scar on examination; 14.9% of the studied infants had negative PPD (0-5 mm) reactions together with absent BCG scars. These findings are significantly different from those previously reported by chilean authors, which showed higher proportion of positive (greater than = 6 mm) reaction to 2 TU PPD in infants from similar populations that had been vaccinated with different BCG preparations than our patients. These results suggest the need to evaluate the efficacy of the BCG vaccines that are currently being used in our country to determine the factors that may affect it and the protection that they afford.

Regional differences in the androgen metabolism were established in alopecic and non alopecic areas of patients affected by male pattern baldness (MPB). 5-alpha-reductase (5-alpha-R) activity was measured by the formation of dihydrotestosterone (DHT), using 3H-testosterone as substrate: this activity was higher in the alopecic areas (3.4 pmol/g tissue/h) than in the non alopecic skin (1.5 pmol/g tissue/h). 3-alpha,beta-hydroxysteroid oxoreductase (3-alpha, beta-HO) was studied using 3H-DHT as precursor and measuring the corresponding formed 3-alpha- and 3-beta-androstanediols (alpha DIOL and beta DIOL). The beta DIOL was the predominant metabolite and total 3-alpha, beta-HO activity was higher in alopecic skin (12.4 pmol/g tissue/h) than in non alopecic areas (8.4 pmol/g tissue/h). Also 17, beta-hydroxysteroid oxoreductase was measured using either testosterone or DHT as substrates: androstenedione formed from testosterone was higher in hairy skin (12 pmol/g tissue/h) than in alopecic areas (6 pmol/g tissue/h); androstanedione formed from DHT was also higher in non alopecic areas (8.1 pmol/g tissue/h) than in alopecic skin (2.8 pmol/g tissue/h). The greater formation of beta DIOL in the sebaceous glands-enriched alopecic skin supports the hypothesis for a specific role of this metabolite in the control of the sebaceous activity.

Dihydrotestosterone (DHT) does not seem to be the active specific metabolite of testosterone in hypertrophic sebaceous glands of subjects affected by male pattern baldness (MPB) and several results indicate that probably 3-beta-androstanediol (beta DIOL) could be an active form of testosterone in those glands. Cytosol and serum from several patients affected by MPB and subjected to hair autotransplantation, was incubated with both beta DIOL and 3-alpha-androstanediol (alpha DIOL). Binding patterns indicate that alpha DIOL binds to cytosolic proteins probably due to the contaminating sex hormone binding globulin (SHBG), whereas beta DIOL exhibits an atypical binding process in cytosol in the presence of high concentrations of non radioactive beta DIOL. This binding increases progressively up to 2 pmol/mg protein at the limit solubility conditions for the non radioactive steroid. This pattern is not observed in serum from the same patients, where the binding of beta DIOL is typically restricted to the SHBG. These results strongly suggest the existence of a specific beta DIOL-binding protein in the hypertrophic sebaceous glands and explain the lack of specific receptor for DHT in these tissues.

The present solubilization strategy recognizes the important role of detergent cocktails in the solubilization and subsequent stability of adenosine A1, receptors from pig brain cortical membranes. The 3-[3-(cholamidopropyl)dimethylammonio]-1-propane-sulfonate-digitonin mixture produced the extraction of up to 52% of the receptor with an enrichment of 1.2-fold with respect to crude membranes. The binding activity of the soluble extract was very stable even in the absence of glycerol. In crude membranes the existence of high- and low-affinity states was detected, but in the soluble extract and in the detergent-treated membranes only the high-affinity state was detected. Association-dissociation curves showed that in crude membranes no interconversion between high- and low-affinity sites is produced by the association of the ligand [3H]R-N6-phenylisopropyladenosine. These results suggest that the high- and low-affinity states are different conformations induced by the structure of the membrane. The modulation of the binding activity by (Gpp(NH)p) 5'-guanylylimidodiphosphate and Mg2+ was studied. In crude membranes Gpp(NH)p shifted the high-affinity state to the low-affinity state, whereas the contrary occurred when Mg2+ was used. The effect of both Mg2+ and Gpp(NH)p was also assayed with the soluble extract and with the detergent-treated membranes. In addition to a decrease of the overall binding capacity, Gpp(NH)p promoted a conversion to all low-affinity states in the detergent-treated membranes or to all very-low-affinity sites in the soluble extract. Mg2+ and Gpp(NH)p counteracted their effects in intact membranes, whereas Mg2+ could not reverse the uncoupling effect of Gpp(NH)p with solubilized or detergent-treated membranes. Thus, it is suggested that Mg2+ acts at sites other than guanine-nucleotide-sensitive sites. If high-affinity states correspond to receptor/G protein complexes and low-affinity states correspond to the uncoupled receptor, we should conclude that Mg2+, as well as the loss of membrane integrity, favours the interaction of A1 receptor molecule with G protein.

Glutathione-S-transferase (GST) activity and glutathione (GSH) content have been studied in human urinary bladder (UB) specimens obtained from healthy controls (HC) (n = 8) and from patients with superficial transitional cell carcinoma (TCC) (n = 9), either in TCC and in adjacent normal (ANE) tissues of the same patient. The GST activity was significantly higher in TCC in comparison with ANE (ten fold) and with HC (five fold). This activity was also significantly higher in HC than in ANE (two fold). The Km values obtained in the whole population (1.26 +/- 0.3 X 10(-3) mol/l) suggest that a unique form of isoenzyme is present in the UB epithelium and that it is the same acidic form "rho" described in erythrocytes. The GSH content was significantly higher in TCC than in ANE (2.5 fold) and also that in HC (three fold). A good correlation between GST activity and GSH content was observed in HC but not in TCC or ANE. These results demonstrate the relation between the activity of the GST system and the development of the TCC as well as its role in the cellular resistance to chemotherapy. A possible decrease of the GST activity before the development of the tumor is also discussed.

The effect of adenosine in rat kidney under normoxic conditions has been studied. It is demonstrated that adenosine modulates cell nucleotide levels. HPLC analysis of the purine compounds inside the cell indicates that adenosine improves the ATP/ADP ratio, whereas it diminishes the adenine content. This behaviour is not due to mediation by specific receptors, as agonists at P1 purinoceptors did not have any effect. Further evidence using inosine as well as dipyridamole and deoxycoformycin indicates that all effects are dependent on the previous uptake of adenosine. The origin of free adenine in the kidney has been investigated, and it appears to come from the phosphorolysis of 5'-methylthio-adenosine. This report is the first to describe the activity of methylthioadenosine phosphorylase (E.C. 2.4.2.28) in the kidney. It is concluded that 1) extracellular adenosine improves guinea pig renal function by increasing the ATP level and the ATP/ADP ratio; and 2) there exists a functional pathway in the kidney that produces adenine and AMP coming from methionine and ATP. This latter pathway probably produces spermine and spermidine, which are likely to be important for renal function.

A quantum chemical study of 10 substrates of adenosine deaminase is performed. The conformational preference around the glycosidic bond of several 8-substituted derivatives of adenosine is studied using semiempirical modified neglect of diatomic overlap (MNDO) and Austin model 1 (AM1) methods. All the compounds studied show preference for the anti conformation; the syn - anti energetic differences calculated are small and in excellent agreement with experimental data. A relationship between the ab initio molecular electrostatic potential minimum energy of N3 and the syn - anti energetic difference is found. A highly significant relationship is also found between the ab initio net charge over the purine and pyrimidine rings and the logarithm of the maximum rate of deamination (log Vm) of the nucleosides by adenosine deaminase. In contrast, no significant relationship is found between the anti preference of 8-substituted derivatives of adenosine and their log Vm of deamination.

Working with pig brain striatum in which A1 and A2 adenosine receptor subtypes coexist, we describe an uncomplicated method for unequivocally obtaining the equilibrium parameters (KD and binding capacity) of A1 receptor without interference from ligand binding to A2 receptor. Also, the equilibrium parameter estimation method we propose avoids the experimental determination of nonspecific binding by the inclusion of the corresponding unknown parameter in the function. This not only saves time but also avoids the use of expensive radioligands in saturation experiments. The method is suitable for any system with two different receptor subtypes for the same physiological ligand, and good estimates of the equilibrium parameters corresponding to the subtype displaying the higher affinity for the ligand can be obtained.

All the boys and girls between six and fourteen years old regularly attending school in three villages from an area in Spain known ass Las Hurdes were studied with respect to thyroid size, body weight and height. Casual urine samples were obtained for the determination of iodine and creatine concentrations. Capillary blood was spotted on filter paper and used for the determination of thyroxine and thyrotropin by radioimmunoassay. The same survey was carried out in 354 school children from Madrid, whose data were used as a reference of the adequacy of the methods employed in Las Hurdes. Goitre prevalence was very high, the overall frequency being 86 per cent. The concentration of iodine in the urine was less than 20 micrograms/1 in 71 per cent of the school children. The concentration of creatine in the urine was also half that found in children from Madrid, and the daily creatine excretion in children from Las Hurdes was lower than expected from their body weight. Their somatic development, as measured by height and weight, was retarded markedly, both as compared to the Madrid reference group and to international charts. Serum T4 was less than 78 nmol/l ( micrograms/kl) in 46 per cent, of the children from Hurdes, and serum TSH greater than 7.5 mU/l in 40 per cent. A relatively low stature (below the 10th percentile) was associated with either a low serum T4, an elevated serum TSH, or both, in 30 per cent of the children. These results indicate that the persistence of endemic goitre in Las Hurdes is of great significance and that there is a continuing risk of the birth of cretins. Observations made a decade earlier in the same area show that the goitre endemic is not diminishing, and that an iodization programme is urgently needed. It also appears that a high proportion of the "normal' schoolchildren have features found in hypothyroidism.

Chloral hydrate is commonly used to sedate infants for pulmonary function tests and other investigations. However, sedation is generally not recommended for infants with acute wheezing illnesses. The commonly used dose range exceeds the maximum recommended dose; however, the effects of this dosage regimen during pulmonary function testing have not been studied. The present study shows that 70-100 mg/kg of chloral hydrate, a dose commonly used to sedate infants for pulmonary function testing, causes a fall in arterial oxygen saturation and a decrease in clinical score of infants recovering from acute viral bronchiolitis, but not in infants with clinically stable cystic fibrosis. These findings suggest that wheezy infants with baseline SaO2 less than or equal to 94% are more susceptible to central respiratory depression following sedation with chloral hydrate. The results of the present study raise serious doubts about the advisability of sedation with chloral hydrate, in the currently used doses, in wheezy infants.

In order to evaluate the clinical usefulness of serum and urinary beta 2 microglobulin (beta 2-m) determination as a marker of renal damage following perinatal asphyxia, twenty asphyxiated and twenty healthy full term newborns were studied. Renal function was monitored on the first and third day after birth by traditional tests such as creatinine (Cr), endogenous creatinine clearance (Ccr), and fractional Na excretion (FeNa), as well as by serum and urinary beta 2 microglobulin. The value of different tests for the diagnosis of oliguria and of acute renal failure was determined. Eleven asphyxiated neonates developed oliguria and five ARF in contrast to none of the controls. Both traditional tests of renal function, and determinations of beta 2-m with the exception of serum beta 2-m, were significantly different (p less than 0.01) between controls and asphyxiated neonates. When stratified analysis was performed, only serum cr, urinary beta 2-m/cr ratio, and Fe beta 2-m were able to discriminate oliguria from preserved diuresis on the first day of life. For ARF, only Ccr and Fe beta 2-m were different, again on the first day of life. Urinary beta 2-m/creatinine ratio and Fe beta 2-m appear to be more sensitive and specific for the early detection of proximal tubular renal dysfunction following perinatal asphyxia than usual tests of renal function.

Synopsis A new formulation for dermatological use and hair care was studied. Phosphate diesters of the nonyl-phenol type in association with cocoimidazoline derivatives are compared with four commercial formulations. Foaming capacity, eye and skin irritation, keratin solubilization and defatting properties were studied. The mixture was the best foamforming formulation and the best defatting agent. Also, it was non-irritant for skin and eyes at adequate dilutions and did not dissolve keratin. It is a good formulation for dermatological use and hair care, due to its detergent properties. It ranks high among safe products.

Female rats were fed a diet with a low iodine content (LID), or the same LID supplemented with KI, and mated. Fetuses were obtained at 17 and 21 days of gestation, or pups were killed at different ages after birth. The dams on LID were markedly iodine deficient and developed a large goiter. Their thyroidal iodine content was only 4% of that of LID + I dams. The iodine deficiency of the LID mothers was severe enough to result in very low plasma T4 levels and in hepatic and cerebral T3 deficiency, despite normal circulating levels of T3. The fetuses from LID dams had low concentrations of iodine in their placentas and thyroid glands, and were deficient both in T4 and T3 in all tissues studied, including the brain. After birth, however, suckling LID pups were able to increase the plasma T4 to levels which were higher than those found in either LID fetuses or in adult LID progeny, although plasma T4 was always lower than in age-paired LID + I animals. This increase in T4 was probably due to an approximately 5-fold increase in iodine intake while suckling. Milk from LID mothers was found to contain 22% of the amount of iodine found in milk from LID + I dams, in contrast to their iodine intake, which was about 4% that of the LID + I rats. Cerebral T3 levels were the same for LID and for LID + I pups throughout most of the postnatal period of brain development. This finding might explain the difficulties encountered in obtaining an experimental model of neurological cretinism in rats.

Myelin was isolated from bovine brain by several published procedures and modifications of these procedures. High activity of the myelin marker (2',3'-cyclic nucleotide 3'-phosphohydrolase) and low activity of contaminants markers in white matter homogenates in respect to cerebral cortex showed the white matter to be better than the cerebral cortex or the whole brain for myelin isolation. A procedure is described for the preparation of purified myelin from bovine white matter which yielded a content of protein (40%), myelin marker (51%), and 5'-nucleotidase (25%) in purified myelin higher than by any used method. Acetylcholinesterase or succinate dehydrogenase was lower than 7% of its activity in the white matter homogenate, and monoamine oxidase and NADPH:cytochrome c reductase were not recovered in myelin fraction. Morphologically, myelin fraction was shown to mainly consist of multilamellar membranes of different sizes. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of myelin fraction showed a characteristic protein pattern of myelin. When our procedure was applied to frozen white matter, lower protein (32%) and myelin marker (34%) and similar 5'-nucleotidase activity (24%) were recovered in myelin, increasing its recovery in denser fractions of white matter.

In order to evaluate the clinical usefulness of serum and urinary beta-2-microglobulin (beta 2-m) determination as a marker of renal damage following cardiopulmonary bypass surgery (CPB), 37 children, with an age range of 6 months to 13 years undergoing CPB under hypothermia and cardioplegia for congenital malformations, were studied. Renal function was monitored at baseline and on the first and third day after CPB by traditional tests such as creatinine (Cr), endogenous creatinine clearance (Ccr), and fractional sodium excretion (FeNa), as well as by serum and urinary beta 2-m measured by radioimmunosorbent assay. Data were analyzed for the group as a whole and after stratification for the presence of acute renal failure (ARF) defined by an increase in Cr greater than 0.5 mg/dl and/or FeNa greater than 2% and of renal hypoperfusion which was considered in case of hypotension resistant to volume repletion and/or dopamine infusion and of oliguria (less than 1.0 ml/kg/h). The incidence of ARF was 18.9%, mortality 21.7% and among those who developed ARF, 71.4%. Although Cr and FeNa increased significantly on the first postoperative day, values returned to baseline thereafter, probably because of the high mortality rate observed among those who developed ARF and were therefore lost to follow-up. In contrast, Ccr was significantly decreased during the whole study. Even though serum beta 2-m displayed a similar profile to Cr and FeNa returning to baseline by the third day, urinary and fractional excretion of beta 2-m were significantly increased at any time of the study.(ABSTRACT TRUNCATED AT 250 WORDS)

The physiopathological role of androgen binding proteins in male pattern baldness (MPB) has been studied by using tritiated dihydrotestosterone (DHT) and methyltrienolone (R 1881) as ligands. DHT binding in bald scalp from subjects suffering from MPB is high (53 +/- 12 fmol/mg protein) in cytosol obtained from bald areas, being undetectable in hairy areas from the same subject. Since methyltrienolone does not bind in bald scalp cytosol, there must be no specific DHT receptor in this material. Several kinetic and molecular parameters of DHT binding in bald scalp cytosol and serum were similar in both samples. Only the association rate constant (k+1) was significantly higher in serum (8.8 X 10(6) M-1 min-1) than in cytosol (3.08 X 10(6) M-1 min-1). DHT binding in serum as well as the evaluation of plasma contamination in the skin samples (by nephelometric analysis) strongly suggests that DHT binding in skin cytosol is merely due to the presence of contaminating SHBG but it does not explain the lack of DHT binding in non bald areas. Thus, the possibility arises of there being a specific mechanism for the uptake of the plasmatic testosterone SHBG-complex taking place only in the hypertrophic sebaceous gland as well as the existence of active T metabolites other than DHT, probably 3 beta-androstanediol.

Purified myelin from fresh calf brain white matter was subfractionated in a discontinuous sucrose gradient; significant recovery of protein and 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNP) and 5'-nucleotidase (5'N) activities occurred in all three obtained subfractions, the highest recovery being in the light subfraction; highest 5'N and CNP specific activities were in medium myelin. Purified myelin was also subfractionated in a continuous sucrose gradient, with a similar localization of protein; CNP activity and 5'N activity maxima suggest that myelin may be a predominant locus of 5'N in bovine brain white matter. Freezing of brain white matter caused an increase in protein and in CNP and 5'N total activity recoveries in denser myelin subfractions. Cytochemistry showed the reaction product of 5'N in the whole myelin fraction to be associated with the innermost, outermost and medial compact myelin layers. Effects of non-ionic detergent (LUbrol WX) on 5'N activity were studied, and the results also suggest the intrinsic nature of 5'N as an ectoenzyme in myelin membranes. Lubrol WX was viewed as an advisable detergent for the stimulation of myelin 5'N activity, but not for the solubilization of this enzyme.

Quantitative and qualitative modifications of the specific binding sites for [3H]quinuclidinyl benzylate (QNB), a muscarinic antagonist, were studied during rat cerebellar postnatal development. Specific binding sites for QNB (QNB-sbs), regardless of whether they correspond to muscarinic acetylcholine receptors, are present with the highest density in the archicerebellar cortex, but the total amount per region is about the same in the archi -, paleo-, and neocerebellar cortex regions. Large amounts of QNB-sbs are also present in a cerebellar fraction including central white matter and deep cerebellar nuclei. QNB-sbs are low but present at birth and then accumulate during ontogenic development according to a curve which duplicates, with a delay of a few days, the curve of DNA accumulation. Dissection studies indicated that this curve does not depend on the preferential localization of QNB-sbs in a specific cerebellar region nor on the particular development of this region. The similarity of the QNB-sbs and the DNA developmental curves might indicate that the QNB-sbs are present on granule cells; however, a comparative analysis of the data in the literature suggests that a great many QNB-sbs are located on the Purkinje cell dendrites in the molecular layer, where all or some of them might correspond to the extrajunctional muscarinic acetylcholine receptor detected there by electrophysiology. It would appear that only a small percentage of cerebellar QNB-sbs corresponds to the cholinergic synapses present in cerebellar cortex; hence, the question of muscarinic receptors in the cerebellum should be re-examined.

Reserpine (0.5 mg/kg i.m.) produced emesis in pigeons with 60% of the animals responding. Metoclopramide HCl at 10, 20 and 40 mg/kg p.o. administered 30 min before or after reserpine injection was effective in blocking reserpine emesis. Metoclopramide was unable to antagonize reserpine-induced sedation and hypotension in rats, thus inviting discussion of its possible mechanism in blocking reserpine emesis.

During the last decade, many new methods of assessing pulmonary function in infants have been introduced. However, considerable controversy has arisen when these different techniques are used to assess the efficacy of various inhaled medications. The currently employed measures of pulmonary function have been reported to have coefficients of variation of less than 10%. However, these figures refer to repeated measurements made in individual babies over a short period of time, with the infant in a constant position. They may not be applicable to studies on the effects of drugs when a significant time interval elapses between measurements, particularly if the infant is repositioned during this time. In the present study, we found that the variability of measurements of thoracic gas volume (TGV), passive respiratory mechanics, and forced expiration approximately doubled if the infant was taken out of the plethysmograph, or repositioned within the plethysmograph, between sets of measurements. The between-test coefficients of variation ranged from 4.8% to 18.9% for TGV, 7.8% to 23.5% for respiratory system resistance, from 5.0% to 24.1% for respiratory system compliance, and from 10.8% to 36.1% for maximal flow at functional residual capacity. These results demonstrate that the commonly employed infant pulmonary function tests have wide variability in infants with acute wheezing illnesses. Before studying the efficacy of therapeutic interventions in such infants, it is necessary to establish the individual variability of the tests in the infants under the conditions of the study. Failure to do so may result in erroneous conclusions and recommendations.

The main purpose of this study was to evaluate four different forms of treatment in young infants admitted for acute wheezing (AW). Seventy-nine infants less than one year of age were randomly assigned to one of five groups. Group 1 received nebulized fenoterol plus ipratropium bromide, group 2 fenoterol, group 3 fenoterol plus steroids, and group 4 aminophylline, IV, plus steroids and oral fenoterol; the control group, or group 5, received nebulized normal saline solution. Clinical evaluation was done by means of a scoring system. The effectiveness of treatments was estimated by a score decrease in the first 24 hours, by the percentage of patients whose scores did not decrease during the same period, and by the number of days in the hospital. All infants had significantly decreased scores, except those in the control group; the aminophylline group included a greater percentage of patients who did not abate their scores, and they stayed in the hospital for more days than those in the other groups. The fenoterol group had the shortest hospital stay. All four treatments produced objective clinical improvement in bronchial obstruction. However, the nebulized bronchodilator treatments were more effective than aminophylline IV in decreasing scores on the first day, and they resulted in shorter hospitalization.

We studied ten patients on hemodialysis (HD) treated in sequence with cuprophan (CU), ethylenevinylalcohol (EVAL), polyacrylonitrile (A-69) and polysulphone (PSP) membrane dialyzers. beta 2-microglobulin (beta 2m) was measured by radioimmunoassay in plasma and dialysate samples. Plasma concentrations were corrected for changes in extracellular volume (ECV). We also studied adsorption in vitro by incubating the above membranes with I-125-labelled beta 2m. There were no changes in beta 2m plasma concentration after HD with CU dialyzers, but a significant decrease was observed with the other membranes tested. Filtration of beta 2m across the dialyzer was absent with CU and minimal with EVAL. However, large amounts were recovered from dialysate with the high-permeability dialyzers, AN-69 and PSP. In vitro studies showed that maximal adsorption capacity was obtained with AN-69 (13%) compared to 9% with CU, 4% with EVAL and 7% with PSP. In summary, beta 2m clearance with PSP is achieved through greater removal of this protein by mass transport across the membrane. The mechanism by which beta 2m is removed from blood during AN-69 dialysis seems to include both adsorption to and filtration by the membrane itself.

The 5'-nucleotidase activity of the purified cytoplasmic fraction preparation of bovine brain does not depend on the presence of the divalent metal ions Mg2+, Ca2+, and Cu2+ in the incubation medium. The Zn2+ ion (0.5 mM) causes total enzyme inhibition. Although EDTA and 8-hydroxyquinoline inhibit the 5'-nucleotidase from this source, it has not been possible to show the existence of metal ions in the enzyme molecule. The inhibition of 5'nucleotidase by EDTA is progressive and irreversible; when the enzyme is not preincubated with EDTA, the inhibition is overridden by metal ions. The purines (except xanthine, 0.3 mM), pyrimidines, and their nucleosides do not affect the 5'-nucleotidase activity. The nucleoside di- and triphosphates are competitive enzyme inhibitors against 5'-AMP as substrate. The Ki values of the diphosphates are lower than those determined for the corresponding triphosphates. The inhibition caused by the above nucleotides is reversed, partly or wholly, by Mg2+, depending on the molar ratio between the effectors. The inhibitory action of the -SH group reagents on the 5'-nucleotidase activity is weak and reversible.