Spontaneous and estrogen-produced tumors in Nb rats and their behavior after transplantation.

Tumors in rats of the Nb strain, arising either spontaneously or after prolonged treatment with s.c. pellets of estrogen, were transplanted to establish whether hormone conditioning was required for their growth. Whereas all spontaneous tumors arising in males and many of those in females were autonomous on transplant, most of those arising in estrogenized rats continued to require hormones for growth after transplantation. The latter included carcinomas of the adrenal cortex, breast, pituitary ectopic tissue, ovary (thecomas), Leydig cells of testis, thymus, pancreas,salivary glands, oribital gland (fibroadenoma), liposarcoma, and lymphoma. Many of the tissues of origin of the tumors have not been considered to be under theinfluence of estrogens. A type of hormone-responsive tumor that was inhibited by estrogen and that grew only in normal rats is described. Ali estrogens tested, including estriol , were interchangeable in action. The incidence of the more common tumors of the adrenal, breast, and pituitary was very low in normal rats, but higher in females. All tumors were more common after estrogenization in both sexes, particularyly in older animals. The secretion of steroids and pitiutary hormones by many tumors led to obvious biological effects. Pituitary secretion led to severe lesions frequently associated with diseases in humans, but the signs of such diseases in the rat apparently were hormone dependent and disappeared if the tumor was removed. The overall results raised the possiblity that estrogens were not carcinogenic per se but stimulated the growth of previously altered cells and that, following their transplantation, this hormone requirement was retained. Irrespective of the mechanism of carcinogenesis, hormone-dependent tumor growth was not irreversible but was controlled in an unexpectedly wide spectrum of organs by exogenous estrogen. Host factors may play a major role in controlling the growth of many tumors and the ultimate course of the disease.


Damage to mammalian cell DNA by nitrofurans.

Maximum rates of nitrofuran reduction by intact mammalian cells and homogenates of mouse liver are obtained under anaerobic conditions, although significant reduction does occur with gas mixtures containing 5% oxygen and less. Single-strand breaks in DNA, measured as a decrease in the sedimentation constant on alkaline sucrose gradients, are produced in mammalian L929, KB, AND BHK-21 cells in vitro and Ehrlich ascites cells in vivo by several nitrofuran derivatives under hypoxic conditons.


In vitro transformation of BHK21 cells grown in the presence of calcium chromate.

Concentrations of 0.25 and 0.5 mug/ml calcium chromate (CaCrO4-2H2O)dissolved in Dulbecco's medium were found to alter the growth behavior of BHK21 cells in culture. Treated cells grew as shortened fibroblasts and in random orientation. The changes detected during the first two weeks of culture in the presence of the metal became more pronounced as the number of growth passages increased. In addition to the alterations noted above, chromate-treated cells grew into large clusters in Methocel (an alternative technique to the agar suspension system), while untreated cells underwent, at most, only one or two divisions in Methocel. These alterations in growth properties were irreversible and persisted after removal of the treated cells from chromate-containing medium, suggesting that a heritable change had occurred as opposed to a transient, chromate-dependent alteration of cell growth. This experimental observation suggests that chromate salts and perhaps salts of other metals can transform BHK21 cells in vitro or can select for spontaneously transformed cells.


Procollagen peptidase: its mode of action on the native substrate.

Procollagen peptidase was recovered from the medium of human and mouse fibroblast cultures by precipitation with ammonium sulfate. The test substrate for the in vitro enzymatic reaction was radioactively-labeled, disulfide-linked procollagen prepared from the medium of human fibroblast cultures. The enzymatic digests were analyzed by electrophoresis in polyacrylamide gets containing sodium dodecyl sulfate and urea. The human and mouse enzymes reacted with the substrate to generate the same intermediates and final products. Procollagen peptidase acts as an endopeptidase which cleaves each of the three procollagen chains in turn. The final products of the reaction are collagen and a three-chain, disulfide-linked fragment derived from the nonhelical aminoterminal residues of procollagen.


Modification of ventricular fibrillation latency following coronary artery occlusion in the conscious pig.

Abrupt occlusion of the left anterior descending coronary artery was performed on 45 unanesthetized farm pigs in order to evaluate the relative effects on the latency to ventricular fibrillation (VFL) of 1) adaptation of the animals to the laboratory, and 2) beta-receptor blockade by propranolol. Compared to control values, VFL was greatly lengthened (i.e., VF delayed or prevented) by adaptation (P smaller than 0.01), was shortened by large (2 MG/KG) doses of racemic propranolol in unadapted animals (P smaller than 0.02), and was again increased while under the influence of the drug, by adaptation (P smaller than 0.051). Neither a lower (0.02 mg/kg) dose of racemic propranolol, 2 mg/kg of dextropropranolol, nor ventricular pacing to a higher heart rate had an effect on VFL. The results suggest that reduced psychological stress was very effective in retarding or preventing the onset of VF, that low doses of propranolol were ineffective, and that higher doses were deleterious in unadapted animals.


Linear working graphs in blood lead determinations with the Beckman flameless atomic absorption cuvet.

Working graphs are generally nonlinear when the longitudinal flameless atomic absorption cuvet is used.  Such nonlinear curves make it difficult to determine the amount of lead in blood by the method of additions, because of inaccuracies in extrapolating the graph to zero absorbance.  This paper describes a method for obtaining linear graphs by making use of opacity (the reciprocal of transmittance, or antilog of the absorbance) instead of absorbance.  Because no deuterium arc background corrector is available for the Beckman Model 444, background was correlated by reading the opacity at a nonabsorbing line, 287.3 nm, and subtracting the opacities.  A separate reagent-blank correction for lead must also be made.


Cultivated cells from diagnostic amniocentesis in second trimester pregnancies. II. Cytogenetic parameters as functions of clonal type and preparative technique.

From a total of 418 primary amniotic fluid colonies, 5.5% were fibroblast-like (F), 33.7% epithelioid (E) and 60.8% had characteristics of what was previously shown to be the principal class of clonable amniotic fluid cells (AF). Polyploidy occurred in all three categories, although both pure tetraploidy and mixoploidy were more frequent in E colonies. The incidence of non-constitutional chromosomal changes was identical in AF and E type colonies if primary spreads were analyzed in situ without prior trypsinization. Spreads from pooled cell suspensions showed higher base-line levels of both aneuploidy and structural changes. Analysis of individual colonies employing an in situ preparative technique is clearly the method of choice for a reliable cytogenetic prenatal diagnosis within the shortest possible period of time.


Parasympathetic pathways, renin secretion and vasopressin release.

1. The interrelationship between parasympathetic neural tone, renin secretion and vasopressin release was examined by observing the effect of bilateral cervical vagotomy on renin secretion in intact and acutely hypophysectomized dogs undergoing a water diuresis. 2. In intact dogs bilateral cervical vagotomy decreased the mean renin secretion from 1245 to 682 units/min (P less than 0.01) as urinary osmolality increased from 95 to 414 mosmol/kg (P less than 0.001). In contrast, in acutely hypophysectomized dogs cervical vagotomy failed to alter renin secretion significantly (834 to 893 units/min) and urinary osmolality was also unchanged (78 to 71 mosmol/kg). 3. The results suggest that a diminution in vagal tone may significantly alter renin secretion by stimulating vasopressin release. Exogenous vasopressin was associated with changes in urinary osmolality and renin secretion which were qualitatively similar to those seen after servical vagotomy. 4. We suggest that there is a neurohumoral reflex mechanism by which a fall in parasympathetic tone increases the release of vasopressin, which, in turn, suppresses renin secretion. The results are also compatible with the hypothesis that vasopressin inhibits renin release by a direct effect on the juxtaglomerular cells.


Deoxycorticosterone excretion in normal, hypertensive and hypokalaemic subjects.

1. Radioimmunoassay has been used to detect and estimate the urinary excretion of deoxycorticosterone (DOC) in normal, hypertensive and hypokalaemic subjects. The range of excretions in ten healthy normal subjects was 41-232 pmol (13.7-76.7 ng) daily, with a mean of 124 pmol (41 ng). 2. In fourteen subjects with essential hypertension without metabolic disturbance the range found was 29-144 pmol (9.7-47.7 ng) daily, with a mean of 87 pmol (28.8 ng), which is not significantly different from that in normal subjects. 3. In twelve patients with Cushing's syndrome due to adrenal cortical hyperplasia the range found was 26-542 pmol (8.7-179 ng). Ten of these twelve patients had values within normal limits. 4. Of nine subjects showing hypokalaemia, eight had elevated excretion of deoxycorticosterone with values from 263 to 5515 pmol (87-1820 ng) daily. Seven of these were hypertensive and two were normotensive. The elevated excretion of deoxycorticosterone found in hypokalaemic subjects is thus not confined to those with hypertension. 5. No correlation has been found between excretion rates for aldosterone and deoxycorticosterone. Raised excretion of the latter provides an indicator of disturbed adrenal cortical metabolism.


Effects of abdominal breathing on distribution of ventilation in obstructive lung disease.

1. The regional distribution of ventilation was studied with a 133Xe wash-out technique both after rebreathing and after intravenous injection of the isotope in six patients with chronic obstructive lung disease. Measurements were performed in the sitting position during spontaneous and augmented abdominal breathing respectively. The contribution to ventilation of the rib-cage and abdomen was measured with magnetometer recordings of the respiratory variation in the antero-posterior diameters. 2. The relative contribution of the abdomen to ventilation increased from about 40% during spontaneous breathing to about 67% during augmented abdominal breathing. 3. There was no systematic difference in the ventilation distribution between the two breathing patterns.


Maximal oxygen uptake, lung volume and ventilatory response to carbon dioxide and hypoxia in a pair of identical twin athletes.

1. Maximal oxygen uptake (VO2 max), lung volumes, and ventilatory responses to carbon dioxide and hypoxia have been measured in identical twin athletes, who were trained to a similar high degree. 2. The results confirm previous findings for VO2 max. and lung volumes in identical twins, and are in keeping with the suggestion that genetic factors play a major part in determining the ventilatory response to carbon dioxide and hypoxia.


Naproxen absorption in children.

A study was carried out in 9 children (mean age 10.8 years) suffering from rheumatic disorders to determine peak plasma levels and plasma curve of naproxen following the administration of a single oral dose of a suspension formulation. The results suggest that there is no significant difference in naproxen peak blood levels and half-life between adults and children and a twice-daily dosage regime, as in adults, should be suitable, therefore, for therapeutic use in children. The formulation was well-tolerated and no untoward effects were noted.


Altered secretion of gonadotropins and steroids resulting from delayed ovulation in the rat.

Rats were killed by decapitation at 3-h intervals during a 48-h delay of ovulation induced by sodium pentobarbital, as well as during the ensuing delayed proestrus, estrus and the first 4 days of gestation. Control animals were killed at the same intervals following injections of vehicle. Blood was collected and analyzed for LH, FSH, prolactin, progesterone and estradiol-17beta to determine if alterations in hormonal levels could account for the abnormal embryonic development which follows delayed ovulation. Hormonal concentrations in plasma were measured by radioimmunoassay except for progesterone, which was determined by competitive protein binding. Embryos were examined to verify the occurrence of abnormal development. Rate of oocyte maturation was studied in serial sections of ovaries from all animals killed at 30-min intervals on the afternoon of proestrus. Oocytes remained in meiotic arrest during the 48-h delay of ovulation, but resumed maturation at the expected time on the afternoon of the preovulatory surge of gonadotropins. Following delayed ovulation there was a significant increase in fertilized ova that were undergoing degeneration (6.5 vs 1.9%), abnormal development (3.7 vs 0.7%), and retarded development (3.1 vs 0.2%). A number of alterations in hormonal levels occurred during the prolonged preovulatory period. The proestrous surge of LH in plasma was completely suppressed on both days of treatment with Nembutal, but exhibited a normal preovulatory peak on the following day. FSH showed a small but significant rise on both days of treatment. Peak concentrations of FSH on the following day were not different in magnitude from the normal preovulatory surge, although the duration of this surge was shortened by 12 h. Prolactin and progesterone concentrations in plasma were only partially suppressed on the days of treatment, while on the following day preovulatory levels of prolactin were significantly greater than in controls. The proestrous peak levels of estradiol occurred prior to the first Nembutal injection and declined more gradually after this treatment. Plasma levels of estradiol remained above basal levels during the period of delayed ovulation with peaks of short duration at 15u0 h on the day of second treatment and the day prior to ovulation. Plasma levels of these 5 hormones during the first 4 days of gestation were not altered by delayed ovulation.


Investigation of choriocarcinoma clonal cell lines in vitro and choriocarcinoma transplants in the hamster for the secretion of a thyroid-stimulating factor.

Six choriocarcinoma cell lines that secrete chorionic gonadotropin (hCG) in tissue culture were investigated for the production of thyroid stimulators. In the mouse thyrotropin bioassay, no thyroid-stimulating activity was found in cell culture media even after 30-fold concentration by the method of Bates. Further, concentrates of culture media did not react in a porcine thyrotropin radioimmunoassay in which hCT cross-reacted completely. Thyroid function was also assessed in hamsters bearing hCG-secreting human choriocarinoma transplants. Even though the thyroid-stimulating factor present in commercial urinary hCG preparations caused an increase in PB131I in the hamster, the presence of the choriocarcinoma had no effect on PB131I. From these results, it appears that human choriocarcinoma cell lines maintained in tissue culture and human choriocarcinoma serially transplanted in the hamster are not suitable models for the study of the secretion of a thyroid-stimulating factor by trophoblastic tissues.


Protein composition of human uterine fluid.

Polyacrylamide gel electrophoresis of uterine washings and the corresponding serum from 35 patients revealed the presence of at least four characteristic uterine proteins and large amounts of the serum proteins, albumin and transferrin. The appearance of these characteristic proteins was studied during specific phases of the menstrual cycle. At lease one of the four uterine proteins was observed in postovulatory samples only, with a maximum frequency of occurrence during the midsecretory phase. In contrast, one of these proteins predominated in preovulatory uterine washings while another appeared randomly throughout the menstrual cycle. Substantial variations in protein patterns were observed in patients sampled during different menstrual cycles. The presence of the major antigenic components of serum in uterine washings was confirmed; however, immunochemical demonstration of the existence of uterine-specific antigens was unsuccessful.


In vitro contractile activity of the mesotubarium superius from the rabbit oviduct in various endocrine states.

We investigated the influence of estrogen on the contractile activity of smooth muscle in the nonpregnant female reproductive tract, using as a model isolated strips of mesotubarium superius (MTS) removed from the rabbit oviduct. We studied four physiologic states in which muscular activity was influenced to varying degrees by endogenous estrogen. Normal estrous (estrogen dominated) rabbits were used as controls. We observed decreased activity in MTS from rabbits chronically deprived of estrogen (60) days postovariectomy). However, a dramatic increase in activity occurred in response to acute withdrawal of ovarian function (12 hours postovariectomy). An increase in activity also occurred at the time of ovulation. We concluded that acute withdrawal of endogenous estrogen is a potent stimulus to reproductive smooth muscle in the female rabbit and that it might contribute to increased muscular activity at ovulation.


Effects of aging on basal levels of serum gonadotropins, ovarian compensatory hypertrophy, and hypersecretion of gonadotropins after ovariectomy in female rats.

Young adult (four months old) and old (14 months old) female rats were used in a series of experiments to determine the effects of aging on serum gonadotropin levels and on ovarian compensatory hypertrophy. Basal levels of gonadotropins were similar for rats of both ages. On the day after hemiovariectomy, serum FSH levels were elevated in both old and young rats. On that same day, serum LH levels were not elevated and the mean level for old rats was significantly lower than that for young rats. At two weeks and at two months after complete ovariectomy, the serum levels of both LH and FSH were lower in old than in young rats, suggesting imparied hypersecretion of gonadotropins in aged animals. After a suppressive dose of estradiol benzoate, ovariectomized old rats showed lower serum LH levels than did young ovariectomized rats. A comparison of the weights of ovaries removed two weeks after hemiovariectomy with those of the ovaries initially removed suggests that compensatory hypertrophy occurred in both groups of rats. However, ovarian weights, both before and after compensation, were lower in the old than in the young. After hemiovariectomy in old rats, the proportion of ovaries with corpora lutea increased, suggesting activation of some previously inactive ovaries.


Temporal evolution of the sensitivity to tetranitromethane of the active site of anti-Tobacco mosaic virus antibodies.

Rabbit anti-TMV antibody taken 9 days after antigen injection is inactivated by much lower concentrations of tetranitomethane (TNM) than anti-body taken after 30 days. This change in sensitivity to TNM occurs in a very heterogeneous population of molecules, including IgM and IgG classes, but the affinity of the antibody remains unchanged. Thus, the phenomenon is not the result of selection of antibody-producing cells under the influence of antigen. Inactivation of early anti-TMV antibody appears to result from the nitration of one tyrosyl residue per Fab fragment. This tyrosine is probably located in the heavy chain and most probably in the variable portion. The moderate sensitivity of late anti-body to TNM is attributed to the lower reactivity of a tyrosine in the active site, and to the fact that nitration of this tyrosine does not inactivate the binding site. The suggested interpretation is that the majority of molecules of a very heterogeneous anti-TMV antibody pool may have a common structure in their active site during the early stage of the antibody response. This may later be replaced in most molecules by different structures.


Immunoelectron microscopic identification and localization of Streptococcus sanguis with peroxidase-labeled antibody: localization of Streptococcus sanguis in intact dental plaque.

Streptococcus sanguis has been localized ultrastructurally within intact dental plaque by means of an indirect technique which utilizes horseradish peroxidase-labeled antibody. The technique allows for complete diffusion of the reagents to all portions of the plaque specimens. Control procedures can be carried out on serial sections of plaque with a bacterial composition similar to that of the experimental specimen. The 30-mum-thick sections can be examined in the light microscope to localize areas specifically labeled with peroxidase prior to cutting ultra-thin sections for electron microscopy. This study demonstrated that specific bacteria can be localized within intact dental plaque. The results also indicated that S. sanguis grows in dental plaque as columnar shaped microcolonies perpendicular to the tooth surfaces. Growth appears to be by cell division rather than deposition of new cells at the surfaces. Despite their relatively good structural preservation, the cells in the deeper (older) layers of plaque appear to have lost some of their antigenic activity in comparison to the cells near the surface.


Application of the cilia dyskinesis test in the diagnosis of cystic fibrosis in an adolescent.

In certain cases of obscure chronic lung disease, the possibility of a genetic disorder, such as cystic fibrosis of the pancreas, should be considered. When the genetic defect fails to cause pancreatic involvement, the partial expression of cystic fibrosis is encountered. Elevation of sweat electrolyte levels is not sufficient to confirm the diagnosis of cystic fibrosis in adults since normal levels vary widely in both adolescents and adults. Other tests should, therefore, be utilized. The cilia dyskinesis test is positive in both homozygous and heterozygous patients with cystic fibrosis. Utilization of this technique in a family study confirmed the diagnosis of partial expression of cystic fibrosis in an adolescent female.


Cyclic hematopoiesis in a colony of dogs.

A laboratory-maintained colony of dogs heterozygous for cyclic hematopoiesis (ch) has been established. The colony consists of Collies and Collie-Beagle mixed dogs. Matings between dogs heterozygous for ch resulted in 110 pups, 28 of which were homozygous for ch. Each of the 28 homozygous ch pups (15 females and 13 males) was gray and white. Ninetten of them survived the preweaning period and were found to have cyclic fluctuations of neutrophils, platelets, and reticulocytes.


Treatment of basophilic leukemia in a dog.

Basophilic leukemia in a 6-year-old dog was characterized by marked splenomegaly, anemia, and leukocytosis in which 89% of the circulating leukocytes were basophilis. After an attempt at busulfan therapy, the dog was treated with hydroxyurea and was maintained on this drug for 2 months. After withdrawl of hydroxyurea, the dog remained in remission and was still in remission at the time of writing (8 months later).


Endonuclease from Dictyostelium discoideum that attacks ultraviolet-irradiated deoxyribonucleic acid.

An endonuclease activity yielding single-strand breaks in ultraviolet light-irradiated phiX-174 RF I DNA in vitro has been detected in homogenates of the amoebae of the cellular slime mold, Dictyostelium discoideum.


Heat stabilization produced by protein-protein association. A differential scanning calorimetric study of the heat denaturation of the trypsin-soybean trypsin inhibitor and trypsin-ovomucoid complexes.

The irreversible thermal denaturation of the association complexes of bovine beta-trypsin with soybean trypsin inhibitor or ovomucoid was observed with a differential scanning calorimeter. Association of trypsin with either inhibitor results in increased heat stability. The largest effect is observed with beta-trypsin and soybean trypsin inhibitor. At pH 6.7, first order rate constants (s-1) for denaturation at 72 degrees, determined at a heating rate of 10 degrees per min, are: beta-trypsin, 30 times 10-3; soybean trypsin inhibitor, 9 times 10-3; trypsin-soybean trypsin inhibitor complex, 0.4 times 10-3. Under equivalent conditions, rate constants for ovomucoid and trypsin-ovomucoid complex are 4 times 10-3 and 1 times 10-3 s-1, respectively. These changes in rate correspond to heat stabilization of trypsin equivalent to an increase of 16 and 9 degrees, respectively, in its observed denaturation temperature. Rate constants determined for beta-trypsin and trypsin-soybean trypsin inhibitor complex are independent of heating rate; those for soybean trypsin inhibitor and ovomucoid are a function of heating rate. This suggests that predenaturational conformational alterations may be important steps in the denaturation of the inhibitors. Activation energies for denaturation of the complexes and their components are all similar, averaging 70 kcal per mol. The large activation energies observed suggest that denaturation of the complexes is not rate-limited by their dissociation.


The participation of lipid-linked oligosaccharide in synthesis of membrane glycoproteins.

Membrane preparations from hen oviduct catalyze the transfer of mannose from GDP-mannose into three components: mannosyl phosphoryl polyisoprenol, oligosaccharide-lipid, and glycoprotein. Eivence that mannosyl phosphoryl polyisoprenol serves as a mannosyl donor for synthesis of both oligosaccharide-lipid and glycoproteins was previously reported (Waechter, C.J., Lucas, J.J., and Lennarz, W.J. (1973) J. Biol. Chem. 248, 7570-7579). In this study the oligosaccharide-lipid has been isolated, and the oligosaccharide has been partially characterized. Based on paper chromatography the oligosaccharide chain contains 7 to 9 glycose units. The glycose at the reducing terminus is N-acetylglucosamine, whereas mannose is found at the nonreducing end. When UDP-N-acetyl[14C]glucosamine is incubated with oviduct membranes in the absence of GDP-mannose, a 14C-labeled chitobiosyl lipid, but little oligosaccharide-lipid is synthesized. When GDP-mannose is also present in the incubation mixture an oligosaccharide-lipid is formed containing N-acetyl[14C]glucosaminyl residues. This oligosaccharide-lipid is chromatographically identical with the [14C]mannose-containing oligosaccharide-lipid isolated in the earlier study cited above. When the N-acetyl[14C]glucosamine-oligosaccharide released from the oligosaccharide-lipid by mild acid is treated with partially purified alpha-mannosidase the major radioactive product is [14C]chitobiose. Evidence that the [14C]mannose-containing oligosaccharide-lipid serves as an oligosaccharide donor for glycoprotein synthesis was obtained by incubation of partially purified oligosaccharide-lipid with the membranes. The products of this incubation were shown to be glycoproteins on the basis of their sensitivity to pronase, as determined by both gel filtration and paper electrophoresis. Similar experiments, using oligosaccharide-lipid doubly labeled with [14C]mannose and N-acetyl[3H]glucosamine, provided evidence that the oligosaccharide chain of the oligosaccharide-lipid is transferred en bloc to glycoprotein s.


The control of adenylate cyclase by calcium in turkey erythrocyte ghosts.

The adenylate cyclase of turkey erythrocytes is inhibited by low concentrations of calcium. Calcium binds to the enzyme system so tightly that the enzyme can compete with ethylene glycol bis(beta-aminoethyl ether)-N, N1-tetraacetic acid (EGTA) for the metal. The calcium binding site is shown to be distinct from the magnesium binding sites required for activity. Thus Ca2+ functions as a negative allosteric effector. Calcium decreases dramatically the V max of the catecholamine-stimulated activity without affecting the affinity for the hormone or for the substrate ATP. The cooperativity in the response toward Mg2+ dependence (Hill coefficient, nH equals 3) is also unaffected by Ca2+ where as the S0.5 (concentration yielding one-half V max) for Mg2+ is affected only slightly. The Ca2+ effect is cooperative (nH equals 2) and therefore brought about by a cluster of Ca2+ binding sites. Mn2+ can substitute for Mg2+ as the enzyme activator but the Mn2+-activated enzyme is no longer inhibited by Ca2+. The possible physiological significance of the Ca2+ effect is discussed.


Identification and purification of collagen-synthesizing polysomes with anti-collagen antibodies.

Antibodies against embryonic chick bone collagen were prepared in rabbits and were purified by affinity and ion exchange chromatography until collagen-specific and RNase-free. 125I-anti-collagen antibodies were used to locate the collagen-synthesizing polysomes of 8-day chick embryo wings and legs on sucrose gradients by measuring the polysome associated radioactivity. The 125I-anti-collagen antibodies bound predominantly to polysomes in the heavy region of sucrose gradients. These binding sites could only be saturated with homologous anti-collagen antibodies. Further evidence for the specificity of this reaction was provided by a correlation of the amount of anti-collagen antibodies bound in the heavy regions of sucrose gradients with the amount of collagen being synthesized by a particular tissue. The validity of this immunochemical method was confirmed by localizing collagen-synthesizing polysomes by an independent method which utilizes their ability to incorporate [3H]proline into collagen peptides in a cell-free system. The collagen-synthesizing polysomes are found in a single, rather broad peak in these gradients. The results of shortening the centrifugation time indicate that larger species of collagen-synthesizing polysomes are not present in these tissues. Partial purification of the collagen-synthesizing polysomes may be achieved by specifically sedimenting them after treatment with anti-collagen antibodies followed by goat anti-rabbit antibodies.


Laser Raman spectroscopic studies of ocular lens and its isolated protein fractions.

The water-soluble proteins of the bovine lens were separated on a column of Sephadex G-200 into five fractions designated as alpha-, beta1-, beta2-, and gamma-crystallin. Laser Raman scattering studies on these isolated proteins (both in the lyophilized state and in solution) and insoluble albuminoid reveal that they contain predominantly antiparallel pleated sheet structure in the main chains and that sulfhydryl groups are highly localized in gamma-crystallin. This light-scattering technique was also applied to probe the homogeneity of protein structure in the intact lens. The analysis of the scattered light selectively collected from various parts of the lens indicated that these proteins also exist in an antiparallel beta structure throughout the entire lens. However, the central (nucleus) and outer (cortex) portions have somewhat different amino acid composition. Based on the relative intensities of the lines at 624 (phenylalanine) and 644 cm-1 (tyrosine), it is concluded that the nuclear part has the highest concentration of gamma-crystallin and that the content of alpha-crystallin increases significantly from the nucleus to the cortex. By examining the Raman spectra in the 2582 cm-1 and the amide I and III regions, we have demonstrated that the sulfhydryl groups and the beta conformation of the lens proteins are unaffected in the conversion of transparent to totally opaque lens by heat denaturation at 100 degrees. This means that the opacification of a lens does not necessarily involve the oxidation of sulfhydrul groups or conformation changes.


Rotational motions in myoglobin assessed by carbon 13 relaxation measurements at two magnetic field strengths.

Proton-decoupled Fourier transform nuclear magnetic resonance spectroscopy of natural abundance 13C was used to obtain spectra of cyanoferrimyoglobin of sperm whale (Physeter catadon) at 14.1 and 23.5 kG. Comparison of the spin lattice relaxation times at these two field strengths allowed the unambiguous assignment of a rotational correlation time of 22 plus or minus 5 ns for the alpha carbon resonances. The spin lattice relaxation time value for a major band attributable to aromatic carbon atoms also corresponded to a single correlation time, attributable to over-all tumbling of the molecule. Certain narrower resonances reflect other modes of rotational motion in addition to the over-all tumbling. Observations of nuclear Overhauser enhancement and line widths accord with these conslusions.


The role of mitochondrial cytochrome P-450 from bovine adrenal cortex in side chain cleavage of 20S,22R-dihydroxycholesterol.

The role of cytochrome P-450 in the side chain cleavage of 20S,22R-dihydroxycholesterol was investigated by examining the effect of carbon monoxide on the conversion of this substance to pregnenolone by cytochrome P-450 from bovine adrenocortical mitochondria; the effect of carbon monoxide on the conversion of cholesterol to pregnenolone by the same enzyme also was examined. Fifty per cent inhibition of side chain cleavage was produced by gas mixtures with the following ratios: CO:O2,1.5 for cholesterol and 1.2 for 20S, 22R-dihydroxycholesterol. Photochemical action spectra revealed that light of wavelength 451 nm decreased the inhibition of side chain cleavage of both substrates to a greater extent than light of other wavelenghts. It is concluded that the heme moiety of P-450 is involved in the cleavage of 20S,22R-dihydroxycholesterol.


Normal plasma renin activity in low renin hypertension.

Plasma renin activity (PRA) was measured every 4 h during a 24-h period of continuous recumbency in 10 patients with essential hypertension. All had maximum values at 12 midnight, 4 AM or 8 AM. Analysis of our date and that of others indicates that in some patients with "low renin" hypertension (LRH) these noctural peaks are of normal magnitude, occasionally exceeding values obtained after four hours of erect posture. Sleep-induced renin release in these patients is not suppressed despite blunted responses to other stimuli. This suggests that the ability to synthesize and release renin may be normal in these patients. Such peaks were not obserived in all LRH subjects, however. The area enclosed by the recumbent PRA curve was also normal in some, but not all patients with LRH. Low renin hypertension may not be a homogeneous disorder.


The thyrotropin in hydatidiform moles is human chorionic gonadotropin.

Thyrotropic activity (TSH), measured by the McKenzie mouse bioassay, has been correlated with human chorionic gonadotropin (hCG) activity, measured by radioimmunoassay, in serum and tissue samples from 11 patients with hydatidiform mole and in partially and highly purified preparations of urinary hCG. Serum samples, taken at various times before and after removal of the moles, gave a ratio of 0.42 plus or minus 0.24 muU TSH/U hCG (mean plus or minus SD) (N)=43). In all cases where hCG activity fell below 150-175 U/ml (n=49), thyroid stimulating activity was undetectable (smaller than 40 muU/ml). We extracted lyophylized molar tissue by a modification of the Bates alcohol-saline method and purified the resultant extract by a combination of gel chromatography, affinity chromatography using Concanavalin A coupled to Sepharose, and isoelectrofocusing. Following extraction, an approximately 20-fold purification was achieved without significant alteration of the ratio of the two activities. Using results from all phases of purification the ratio of muU TSH/U hCG was 0.51 plus or minus 0.35(n = 23). Both activities were in the same position on disc gel electrophoresis. Activity ratios were less constant when partially purified preparations of urinary hCG were assayed for both thyrotropic and hCG activities. The presence of an hCG immunoreactive species, presumably hCG-beta subunit, which contains no thyrotropic activity but has an approximately 10-fold greater activity on a weight basis than intact hCG, may be a partial explanation for this observation. Isoelectrofocusing of a urinary hCG preparation showed that all hCG immunoreactive species with pl's between 3. 5 and 5.0 contained thyrotropic activity in proportion to their hCG content. Seven highly purified hCG preparations had thyrotropic activity with a ratio of 0.48 plus or minus 0.18 muU TSH/U hCG. These results indicate that hCG has intrinsic thyrotropic activity. On a molecular basis it is calculated that hCG contains approximately 1/4000 the thyrotropic activity of human pituitary TSH. In conditions of grossly elevated serum hCG levels, such as hydatidiform mole, this thyrotropic activity can be sufficient to produce hyperthyroidism.


pH dependence of acid secretion and gastrin release in normal and ulcer subjects.

By use of a recently described method, which estimates the rate of gastric acid secretion by measuring the rate of sodium bicarbonate infusion needed to keep intragastric pH constant, gastric acid secretion rates and changes in serum gastrin were measured in five normal subjects while gastric pH was kept at 5.5, 4.0, 3.0, or 2.5. Preliminary experiments revealed that the method did not accurately measure acid secretion at a pH lower than 2.5. Stimulation of acid secretion was produced by gastric instillation of a solution of amino acids and cornstarch. The secretion rate with the amino acid meal was highest at pH 5.5 and was 60% of that produced by a steak meal at the same pH. As the pH of the amino acid meal was decreased, there was a stepwise reduction in acid secretion so that at pH 2.5 the rate was only half as great as at pH 5.5. The amino acid meal produced increases in serum gastrin that were also less marked than those produced by a steak meal. With amino acid stimulation, serum gastrin responses were similar at pH 5.5, 4.0, and 3.0, but no increase in gastrin could be measured when the meal was maintained at pH 2.5. A group of six patients with duodenal ulcers was compared with seven normal subjects at pH 5.5 and 2.5. Ulcer patients released more gastrin and secreted more acid at each time period at both pH values. More important, the degree of inhibition at pH 2.5 was significantly less in ulcer patients. For example, during the 2nd h after stimulation acid secretion was inhibited by only 30% in ulcer patients compared with 70% in normal subjects. These findings suggest a defect in autoregulation of gastrin release and gastric acid secretion at low pH in ulcer patients which may play a role in pathogenesis of this disease.


Globin synthesis in fractionated Normoblasts of beta-thalassemia heterozygotes.

Globin chain synthesis was examined in erythroid cells of increasing maturity, fractionated from the whole bone marrow of beta-thalassemia heterozygotes by a density gradient centrifugation procedure. In experiments using total cell "globin," a gradient of alpha/beta chain ratios was observed, increasing with erythroid cell maturation from unity in the basophilic cells up to 2.0 in reticulocytes. Gel filtration of the lysates from these marrow fractions revealed the presence of free alpha chains even in the most immature cells, the amount of which increased with erythroid cell age; the total alpha/beta ratio derived from gel filtration experiments showed a gradient similar to that observed in the total globin experiments. However, the alpha/beta ratio of the hemoglobin fraction obtained by gel filtration remained constant throughout maturation at an average of 0.65. This latter finding is incompatible with balanced synthesis at any stage of red cell development and excludes the possibility that total beta chain production is higher in the early cells than in the later cells or that alpha chain production in the early cells is reduced to the level of beta chain synthesis. Furthermore, in a Hb S/beta-thalassemia marrow examined, the beta A/beta S ratio remained constant throughout maturation while the alpha/non-alpha ratio showed an increase like that observed in the simple beta-thalassemia heterozygotes. This argues strongly against increased synthesis from either the thalassemic or nonthalassemic beta chain gene being responsible for the balanced synthesis in the immature cells. These findings lead us to suggest that, in beta-thalassemia heterozygotes, a large alpha chain pool is present throughout erythroid cell maturation and that the observed increase in alpha/beta ratios is a function of the ability of those cells to degrade the excess alpha chains.


Chemotactic activity derived from interaction of factors D and B of the properdin pathway with cobra venom factor or C3B.

Interaction of D (the activated form of D) and B, factors of the properdin pathway, with C3b (the major cleavage fragment of C3) generates a convertase, C3B, which cleaves C3 and initiates the terminal complement sequence C5-C9. A functionally analogous more stable C3 convertase, CoVFB, ir formed by substituting cobra venom factor (CoVF) for C3b. Mixtures of highly purified CoVF, B, and D were chemotactic for human neutrophil polymorphonuclear leukocytes as assessed in Boyden chambers either by microscopic enumeration of migrating cells or by counting of 51Cr-labeled cells. Control mixtures containing CoVF, B, and D, reacted in the absence of Mg++, were hemolytically inactive and devoid of chemotactic activity. Over a range of doses, the chemotactic activity of mixtures yielding CoVFB correlated with their hemolytic activity. Pretreatment of neutrophils with mixtures containing CoVFB rendered them unresponsive to subsequent chemotactic stimulation by kallikrein of C5a, indicating cross-deactivation to other chemotactic factors. Similar neutrophil deactivation occurred after exposure to a mixture of C3b, B, and D in which C3B was formed; with short incubation times and high cell concentration C3B also exhibited some chemotactic activity. The chemotactic activity of C3B and CoVFB is an example of a biologic function arising from interactions among factors of the properdin pathway per se, as distinguished from the capacity of this pathway to activate C3 and the terminal complement sequence.


Potential sources of copper contamination of farm milk supplies measured by atomic absorption spectrophotometry.

A method was developed to use atomic absorption spectrophotometry for copper measurements in milk filtrates prepared from equal volumes of milk and 20% trichloracetic acid. Results are reported for 11 farms for water supplies, wash and rinse solutions, and milk. Copper below 10 mug/liter was found at the well for the 5 farms with their own spring supply. Averages and ranges for all 11 farms in mug/liter were 33 (0 to 156) at the cold water tap in the milk house, 220 (8 to 1,360) at the hot water tap, 246 (8 to 1,722) in the wash sink with added detergent and hot water, and 275 (9 to 1,823) after using the wash solution. Rinse solution with hypochlorite in cold water showed a rise from 31 (1 to 157) before use to 51 (5 to 206) after use. Copper in milk averaged 99 mug/liter (72 to 163). In spite of relatively high copper in wash solutions, copper in milk was not correlated postively, giving rise to question whether adsorbed copper from such solutions can contribute much copper to milk. Thorough rinsing with low copper cold water and a high dilution ratio in milk were seen as mitigating factors. Direct contact of milk with copper bearing metals presented a greater opportunity for contamination. Removal of substantial amounts of copper from pipe lines and bulk tanks with sulfamic acid was demonstrated.


The histochemical demonstration of hydrolase activities with films of homopolyribonucleotides.

Films of polycytidylic acid, polyuridylic acid and polyguanylic acid were exposed to tissue sections and the results were compared with those obtained in previous studies on polyadenylic acid and ribonucleic acid. Important variations were observed in the distribution of the hydrolases acting on the different polyribonucleotides, suggesting that a variety of nucleases with marked proclivity for particular nucleotide residues can be demonstrated by the use of films of homopolymers.


The cytochemical localization of succinic dehydrogenase in the sperm mitochondria of the gastropod Biomphalaria glabrata. A comparison of two methods.

The mitochondrial derivative of the sperm of the gastropod pulmonate Biomphalaria glabrata was studies to ascertain succinic dehydrogenase localization cytochemically. Two techniques were compared. One technique depends on a tetrazolium salt that yields an osmiophilic formazan upon reduction. The other technique is dependent on the reduction of copper ferricyanide. The effects of several electron transport inhibitors were studied. The reaction product observed in the matrix of the mitochondrial derivative using the former technique is sensitive to rotenone and is believed to be nicotinamide adenine dinucleotide-dependent. The reaction product observed in the intracristal spaces using the copper ferricyanide method is insensitive to rotenone and is believed to cytochemically demonstrate succinic dehydrogenase in this material.


Localization of a thrombin-sensitive calcium pool in platelets.

Electron microscopic x-ray microprobe analysis of pyroantimonate precipitates in platelets fixed in osmium tetroxide-pyroantimonate revealed calcium localization in the nucleoids of alpha-granules. This pool of calcium had largely disappeared within 10 sec after stimulation of platelets by thrombin. Such a rapid change suggests that this calcium pool may have a regulatory role in stimulus-response coupling.


Dipotassium tetramethyl osmate: a stain for electron microscopy.

Methanol solutions of dipotassium tetramethyl osmate (DTMO) have been found to be useful as general stains in electron microscopic studies of plant and fungal ultrastructure. The stain solutions are easy to prepare, stable when anhydrous and convenient to use. Although generally similar in staining to lead citrate stains, some elements of cell ultrastructure appear different with dipotassium tetramethyl osmate staining, particularly the outer cell walls of fungi. Indications of specific precipitate-producing reactions in cell storage areas are observed.


An expression for the calculation of relative affinities of antibody-ligand interactions.

An expression is derived which allows the calculation of K-a without a value for total binding sites of antibody. The expression yields unambigous K-a values for homogeneous populations of antibody molecules. When heterogeneous populations are used, the value of calculated K-a rises as the fraction of sites bound falls. This expression should  be useful in the determination of relative affinity with data obtained from Farr assays of the binding of anti-hapten antibody to hapten. Since values for total binding sites are unnecessary, this equation should be expecially applicable for the study of heterogeneous populations of relatively low affinity antibodies present at low concentrations.


Immunochemical quantitaion of the third component of guinea pig complement in fluid phase and bound to cell surfaces.

A method is described for the determination of the absolute amount of guinea pig (GP) C-3 bound to cell surfaces. The test is based on the inhibition of anti-C-3 by C-3. The C-3 to be measured is added to a standard amount of anti-C-3 antibody and the residual activity is determined by its capacity to sensitize EC43-gp for lysis by GPC. By comparing the observed lysis with inhibition-of-lysis curves obtained with known C-3 antigen, the amount of test antigen can be determined. The method is simple, rapid, and reproducible and can detect nanogram amounts of C-3. The C-3 content of six normal guinea pig sera was found to be 1365 plus or minus 56 mug/ml (ranges 1120 to 1510 mug/ml) by the inhibition method described in this paper. Guinea pig hepatoma cells, line-1 and line-10, harvested from the peritoneal cavity of guinea pigs contained approximately 6.7 x 10-3 to 6.2 x 10-4 GPC-3 per cell whereas tumor cells sensitized with anti-Forssman or specific anti-tumor antibody contained from 1 to 9 x 10-5 C-3 per cell. Susceptibility to killing by antibody and GPC could not be correlated with the amount of GPC-3 bound to the surfaces of the tumor cells. Comparisons made between the amounts of C-3 bound to antibody-sensitized sheep erythrocytes and nucleated cells suggest there is a fundamental quantitative difference in the fixation of selected C components.


Complement receptor in synchronized cultures of human hematopoietic cell lines.

Three previously established human lymphoid cell lines (HLV-1, SWB-16A, and B-1) were synchronized by double thymidine block and the lymphocyte surface receptor for complement was studied by the EAC-rosetting technique during the cell cycle. It was found that the proportion of cells with EAC rosettes remained unchanged throughout the various stages, indicating that the presence of the complement receptor in cultured lymphoid cells is independent of the cell cycle.


Immunologic and physicochemical properties of enhancing soluble factors for IgG and IgE antibody responses.

Rabbits were immunized with dinitrophenyl-coupled Ascaris antigen (DNP-Asc) or ragweed antigen (DNP-Rag) included in aluminum hydroxide gel and their mesenteric lymph node cells were cultured for 24 hr in vitro in the presence of free homologous carrier. The cell-free supernatant thus obtained enhanced both IgG and IgE antihapten antibody responses of DNP-primed cells to DNP-heterologous carrier conjugate (DNP-keyhole limpet hemocyanin). Since the cell-free supernatant obtained from Rag-specific cells enhanced antibody response of hapten-primed cells raised by immunization with DNP-Asc, no carrier specificity was involved in the enhancement. It  was found that treatment of primed cells with 10-5 M pactamycin suppressed the formation of the enhancing soluble factor, whereas the factor was readily formed in the presence of 2 mug/mol of  cytosine arabinoside in the culture. The results indicated that cell proliferation was not required but de novo synthesis of protein was essential for the formation of soluble factor(s). The enhancing factor  was not absorbed by either carrier-coated or anti-carrier antibody-coated immunosorbent. It was also found that the enhancing factor was formed by incubating primed cells with carrier-coated Sepharose. The cell-free supernatant containing no free carrier enhanced both IgG and IgE anti-hapten antibody responses. The activities of the cell-free supernatant to enhance IgG and IgE antibody responses were not absorbed by anti-Fab, anti-gamma-or anti-mu-chain antibody immunosorbent, indicating that the nonspecific enhancing factor did not possess immunoglobulin determinant. The cell-free supernatant was fractionated by sucrose density gradient ultracentrifugation and by gel filtration with three radiolabeled proteins, i.e., IgG, ovalbumin, and cytochrome C as markers. Enhancing activity for IgG antibody response was recovered in a fraction between ovalbumin peak (40,000 m.w.) and cytochrome C peak (20,000 m.w.). The activity for IgE antibody response was recovered in a fraction containing IgG marker (150,000 m.w.). By block electrophoresis, both activities were detected in beta globulin fraction. The results suggested that different T cell factors are involved in the IgG and IgE antibody responses.


A study of the association of human secretory component with IgA and IgM proteins.

Human secretory component (SC) was isolated from colostral whey, and the binding of 125I-SC to purified IgA and IgM monoclonal proteins was studied using two methods to separate free from immunoglobulin-bound 125I-SC: a) gel filtration on Sephadex G-200, and b) precipitation of 125I-SC-Ig complexes with anti-Ig antibody. Both IgA dimeric proteins and IgM pentamers bound 125I-SC with approximately one SC-binding site per mole of polymer and similar affinity. Assuming a reversible equilibrium, an apparent association constant congruent to 10-8 M-1 was calculated to govern the binding of 125I-SC to immunoglobulin polymers. The assignment of a single association constant may be an oversimplication, particularly for the case of IgA polymers, since evidence was obtained that disulfide bonds were formed in the 125I-SC-IgA complex. Despite the complexity of the reaction, binding of 125I-SC to both IgA and IgM polymers could be analyzed by standard methods of saturation analysis, and both were shown to have a similar affinity for 125I-SC. No differences were noted in the affinity of 125I-SC binding to the IgA1 and IgA2 subclasses. Binding of monomeric IgA and IgM proteins could not be measured and was at least 100-fold lower than that found for IgA and IgM polymers. Complexes of 125I-SC with IgA dimers were presumed to involve covalent bond formation, since these complexes did not dissociate in guanidine-HCl. One IgA2 trimer did not form a covalent bond since it was completely dissociated in guanidine. In contrast, 125I-SC-IgM complexes were dissociated in denaturing solvent, indicating that such complexes were held together primarily by non-covalent bonds. Experiments with (Fc)5 mu isolated by high temperature tryptic digestion of IgM showed that binding of 125I-SC was to the Fc region of IgM proteins. It was suggested that the binding of SC with similar affinity to both IgA and IgM polymers may be important in the biologic function of both these immunoglobulin classes.


Purification of DNP specific antibodies using sepharose bound DNP-para-aminobenzoylglutamate.

An immunoabsorbent column employing a DNP derivative having restricted molecular freedom (dinitrophenylated-para-aminobenzoylglutamate--Sepharose 4B) was prepared in order to isolate and purify DNP specific antibodies in high yield. The antibodies were recovered from the immunoabsorbent column in yields from 80 to 90% using 3 M sodium thiocyanate, and these antibodies retained their immunologic activity. There appeared to be limited selection of antibodies with specific affinities as was noted in a parallel experiment comparing a second antibody purification procedure.


Comparison of scrapie and transmissible mink encephalopathy in hamsters. II. Clinical signs, pathology, and pathogenesis.

Scrapie and transmissible mink encephalopathy were studied in hamsters; clinical signs, pathology, and the replication of the agents of each disease in brain and spleen were compared. The most noticeable clinical sign in scrapie-affected hamsters was a distinct cerebellar ataxia beginning 16 weeks after inoculation. Ataxia was not prominent in animals affected with transmissible mink encephalopathy; these animals gradually became more and more lethargic. The pathology in the central nervous system in both diseases consisted of astrocytic hypertrophy, microvacuolation of the neuropil, and neuronal degeneration. The scrapie agent appeared to have a greater effect on nuclear masses, especially those present in brain stem and the central white matter of the cerebellum. The earliest lesions in both diseases were detected near pia arachnoid surfaces and adjacent to the ventricular system. These initial sites of involvement suggest that the cerebrospinal fluid may be an important route by which inocula are disseminated to susceptible cells after intracerebral inoculation. Both agents multiplied rapidly in brain, reaching titers greater than 10-8 ld-50/0.05 ml before the onset of clinical signs. Titers in spleen were 4-6 logs lower than titers in brain at every point measured during the asymptomatic or clinical course of disease.


Comparison of scrapie and transmissible mink encephalopathy in hamsters. I. Biochemical studies of brain during development of disease.

A series of 15 different biochemical measurements were made on brains taken at stages throughout the development of scrapie and of transmissible mink encephalopathy in hamsters. With both diseases biochemical abnormalities were found only after the development of early histologic lesions, when animals showed clinical signs of disease. Changes were recorded in body weight, in the activities of six glycosidases, and the rates of incorporation of DNA and of glycoprotein precursors. The profiles of changes in hamster brain were almost indistinguishable,qualitatively and quantitatively, in the two diseases, an  observation suggesting a close similarity in the way both disease-producing agents interact with this particular host species, . However, there were some major differences between the profile of changes in hamster scrapie and that previously observed in mouse scrapie. Thus it would appear that many of the well-characterized abnormalities of mouse scrapie are not fundamentally involved in the development of disease but represent mainly secondary changes.


A sensitive, rapid, and economic radioimmunoassay of human growth hormone using ethanol-ammonium acetate.

An aqueous solution of 66 per cent ethanol and 6.6 per cent ammonium acetate has been used in the development of a relatively simpler and rapid as well as highly reproducible method of separation of antibody-bound and free hormones in the radioimmunoassay of human growth hormone (HGH). The present assay is sensitive to detect 20 pg. of hormone with a precision of plus or minus 5 per cent, hence, as little as 20 mul of plasma is required to perform the radioimmunoassay. Serum levels of HGH have been found to be approximately 70 per cent of the plasma concentrations in the same subjects.


Relationship of the sex of the foetus to the amount of human chorionic gonadotrophin in placentae; single and dizygotic twin placentae compared.

Human chorionic gonadotrophin (HCG) was assayed by biological and radioimmunological methods in placentae from 16 women with a normal twin pregnancy. When the concentration and total amount of HCG in placentae was related to the sex of the twin foetus, no significant difference between 'male' and 'female' placentae was found. This is contrary to findings that there is a significant (P less than 0-005) difference in the concentration of HCG per g and per placenta of singletons at term. A comparison between the grouped geometric mean data from bioassays shows that the amount of HCG per g and per placenta falls between the geometric mean values for 'male' and 'female' singleton placentae.


Relatively greater increase of oxytocin than vasopressin in the supraoptic nucleus of rats after hypophysectomy.

The vasopressor and milk ejection activities were estimated in the supraoptic region of the hypothalamus of rats which had been hypophysectomized 30 min, 4 and 8 days earlier. Both pressor and milk ejection activities were significantly greater 4 days after hypophysectomy than those in sham-operated control rats. Eight days after hypophysectomy, pressor activity decreased below control values but milk ejection activity was still significantly raised. Increased biological activity after hypophysectomy is thus associated with the previously observed increase in neurosecretory material within the supraoptic nucleus so these results provide additional evidence that neurosecretory material represents stored hormone. Milk ejection activity increased relatively more than pressor activity which supports the suggestion that the final stage in the formation of neurohypophysial hormones occurs as they pass from the hypothalamus to the neural lobe. If this final maturation normally occurs more slowly for oxytocin than for vasopressin, the obstruction to the flow of hormones down the neural stalk caused by hypophysectomy would result in a greater increase in milk ejection than pressor activity.


Effects of postpubertal oestrogen injections on mitotic activity of vaginal and uterine epithelial cells in mice treated neonatally with oestrogen.

Cell cycles of vaginal and uterine epithelial cells were studies using [3H]thymidine autoradiography in adult ovariectomized mice given oestrogen injections neonatally. The mice were in a 'persistent-oestrous' state, showing ovary-independent, continued proliferation and cornification of the vaginal epithelium. The duration of different stages of the cell cycle could not be assessed in such mice, since the percentage of labelled mitoses failed to rise to 100%. In neonatally oestrogenized, adult mice the vaginal epithelium appeared to contain a mixed population of cells. After an oestrogen injection, almost all mitoses of vaginal epithelial cells became labelled, with a generation time of about 17 h. By contrast, the generation time was about 15 h in vaginal epithelial cells of ovariectomized 'normal' mice injected with oestrogen when adult. The uterine epithelium of neonatally oestrogenized, ovariectomized mice also consisted of a mixed population of cells. A single oestrogen injection produced an increase in both the mitotic rate and cell number in the vaginal and uterine epithelium of ovariectomized 'normal' adults but not in neonatally oestrogenized, ovariectomized adults. These studies show that in mice given oestrogen neonatally, uterine and vaginal epithelial cells were not responsive to oestrogen or at least less sensitive to oestrogen than ovariectomized 'normal' controls.


Levels of follicle-stimulating hormone, luteinizing hormone, oestradiol-17 beta and progesterone, and follicular growth in the pseudopregnant rat.

Throughout a period of pseudopregnancy the peripheral blood levels of progesterone, oestradiol-17 beta, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), as well as the size-distribution of ovarian antral follicles were estimated in the rat. The progesterone concentrations, as measured by a competitive protein-binding technique, exceeded metoestrous values (25 ng/ml plasma) from day 3 of pseudopregnancy onwards. The highest levels were found on days 6 and 8 (91 ng/ml). From day 8 onwards the levels decreased gradually but were still above metoestrous values on the day of pro-oestrus after pseudopregnancy. Concentrations of oestradiol-17 beta, as measured by radioimmunoassay, were within the range of those at metoestrus (about 5 pg/ml plasma) until day 10. Thereafter levels increased to a value of 57 pg/ml. Concentrations of FSH, measured by radioimmunoassay, were within the range of metoestrous values until day 10 (about 100 ngNIAMD-rat-FSH RP-1/ml serum), but declined to a level of 33 ng/ml on day 12. Concentrations of LH, measured by radioimmunoassay, were generally within the wide range of metoestrous values (9-60 ng NIAMD-rat-LH RP-1/ml serum), but concentrations found on days 4, 8 and 10 were significantly lower than those found on preceding or subsequent days. Histological determination of the number of follicles present in various volume-classes, showed an increase in antral follicles on days 1 and 2, comparable to the increase observed during metoestrus and dioestrus 1 of the normal cycle. There was no change in the follicles between days 3 and 10 and they resembled those of early dioestrus. Preovulatory growth had occurred by day 12. Injection of human chorionic gonadotrophin (HCG) on days 2, 4 or 6 showed that ovulation could be induced only in some of the larger follicles. On the basis of these results it is suggested that during pseudopregnancy the high progesterone levels present result in a decreased plasma LH level which is insufficient to cause full maturation of the follicles and to stimulate oestrogen secretion to the levels required for induction of an ovulatory surge of LH release.


The influence of serum on the uptake, conversion and action of dihydrotestosterone in rat prostate glands in organ culture.

The influence of serum on the uptake, conversion and action of dihydrotestosterone in relation to the sex steroid binding protein, TeBG, has been investigated in rat ventral prostates in organ culture. The organs were incubated with (1,2-3H)dihydrotestosterone in: (1) serum-free medium, (2) horse serum, foetal and newborn bovine serum of (3) human male and human pregnancy serum. With all sera the uptake of dihydrotestosterone fell with rising serum concentration, at first steeply and then more gradually. At the same concentration, the uptake was significantly lower in explants incubated with human pregnancy serum than in those kept with human male serum. The conversion of dihydrotestosterone to androstanediol followed the same pattern and less androstanediol was formed in the presence of pregnancy serum. Since pregnancy serum contains higher amounts of TeBG than male serum, the lowered uptake suggests that only the free hormone was available to the target organ. Addition of unlabelled dihydrotestosterone resulted in a higher uptake than that measured in explants incubated with the labelled steroid only. The effect of the human sera on uptake and conversion was correlated with the androgenic activity of dihydrotestosterone applied at physiological concentrations and expressed as the percentage of secretory columnar cells present. The degree of maintainance closely corresponded to the uptake of the hormone. In serum-free medium, the number of columnar cells approached the values found in vivo, with male serum their number, though reduced, was still substantial, with pregnancy serum it was extremely low. It is concluded that the amounts of TeBG present in serum regulate the supply of the hormone to the target tissue and thus control its biological action.


The effects of dexamethasone and indomethacin on the outcome of pregnancy in the rabbit.

Pregnant rabbits were treated with indomethacin (8-10 mg/kh/day) or dexamethasone (1-2-1-8 mg/kh/day) during late gestation. The effects of these treatments on the concentrations of progesterone and prostaglandin F (PGF) in the peripheral plasma, and the outcome of gestation was studied. Treatment with indomethacin significantly prolonged the length of gestation (P smaller than 0-01) compared iwth control, untreated animals. In these treated animals, the plasma progesterone levels declined at a similar time to that in control rabbits but the increase in systemic PGF normally seen during late pregnancy was reduced. Dexamethasone treatemnt reliably induced premature delivery within 3-6 days. The plasma progesterone concentration fell rapidly during the first 24 h of dexamethasone administration, but in no animal was this associated with a significant increase in the plasma levels of PGF. These results are consistent with the suggestion that prostaglandins are involved in the normal initiation of parturition in the rabbit. They do not support the hypothesis that the effect of dexamethasone on the length of gestation is mediated through an increase in the production of prostaglandin F.


Frequency of reinforcement as a determinant of extinction-induced aggression during errorless discrimination learning.

Seven pigeons were trained to discriminate without errors between a green keylight and a dark key. The key-pecking response was reinforced in the presence of green, and extinction was in effect in the presence of the dark key. The opportunity to attack a restrained target pigeon was present only during extinction. Both variable-interval 30-sec and fixed-ratio 1 schedules of reinforcement during the positive stimulus induced a higher rate of attack during extinction than a variable-interval 5-min schedule. The highest rate of attack during extinction occurred during the first 20 sec after the positive stimulus terminated. Hence, the withdrawal of the positive condition, rather than the consequences of the pecking response during extinction, appears to be one of the primary factors responsible for attack between pigeons during extinction. Behavioral contrast, defined as a decrease in the rate of responding when the positive stimulus was presented alone, was obtained from the four birds that displayed the lowest overall rates of attack while the three birds with the highest attack rates did not display behavioral contrast. For the birds without contrast, components of the attack response during the positive stimulus presumably competed with and reduced the rate of pecking the key, thereby recluding behavioral contrast.


A simple, rapid 125I radioimmunoassay for the detection of barbiturates in biological fluids.

A simple, rapid radioimmunoassay employing 125-I-labeled secobarbital derivative has been developed and has been shown to be capable of detecting at the nanogram level a variety of barbiturates in urine as well as in plasma.


The fine structure of staggerer cerebellum.

The cerebellar cortex of staggerer mice, ranging in age from 12 days to three months, was studied in the electron microscope. In general, the cerebellum showed a much delayed developmental pattern with regard to Purkinje cell spine formation. Spines did not appear on the soma until about 6 days after birth and appeared on the dendritic branchlets at only 23 days. Climbing fibers were present and they were in synaptic contact with both somatic and dendritic spines. By the third month, spines were only rarely obserbed and some of these were unattached to any presynaptic element.


Spontaneous intracerebral hematoma caused by cerebral neoplasms. Report of eight verified cases.

In eight of 590 consecutive patients operated on by the author for a proven brain tumor, the preoperative diagnosis was complicated by a spontaneous intracerebral hematoma caused by the neoplasm. The presenting symptoms were those of hypertensive intracerebral hemorrhage or brain tumor. The pathology underlying spontaneous bleeding from a cerebral neoplasm is reviewed and diagnostic suggestions discussed.


Utilization of nitrogen from soybean meal, casein, zein, and urea by mature sheep.

The utilization of nitrogen contained in soybean meal, casein, zein, and urea was studied in 12 mature wethers. Net microbial synthesis rates during the 6-hour period after feeding corresponded to ruminal ammonia concentrations, suggesting that the ruminal ammonia level in these experiments was a limiting factor in microbiol protein synthesis. Maximum conversion of dietary nitrogen to microbial nitrogen in the rumen during a 24-hour period was estimated to be 62.5, 85.3, 40.9, and 90.1%, respectively, for soybean meal, casein, zein, and urea. Estimated production rates of acetic, propionic, butyric, isovaleric, and valeric acids during the first 6 hours after feeding suggested that zein supported the lowest microbial activity in the rumen, and that urea supplementation resulted in isovaleric and valeric acids production rates equivalent to or greater than rates when the other nitrogen supplements were present in the diet. Daily endogenous urinary and metabolic fecal nitrogen determined by regression analysis utilizing soybean meal as the only nitrogen supplement were 72 mg/kg body weight0.75 and 340 mg/100 g of dry matter intake. Biological values of 84.8, 78.3, 84.3, and 76.4 were obtained for soybean meal, casein, zein, and urea, respectively.


Acetylmethionine as a source of methionine for the rat.

A-Acetyl-L-methionine and N-acetyl-D-methionine were compared with L-methionine and D-methionine as sources of methionine. These derivatives were added to a sulfer amino acid-limited diet containing 10% soybean protein isolate. Weight gains, food intake, and protein efficiency ratios (PER) were determined in growing rats. N-Acetyl-L-methionine, L-methionine, and D-methionine produced an equivalent growth response and increase in PER above that of the basal diet. There was no response to supplementation with N-acetyl-D-methionine. An equivalent maximum growth response of rats fed-L-methionine or N-acetyl-D-methionine. An equivalent maximum growth response of rats fed L-methionine or N-acetyl-L-methionine was obtained when the total dietary sulfur amino acids compromised 0.36-0.41% of the diet. The nutritional similarities of methionine and N-acetyl-L-methionine suggest that the latter may be useful as a supplement to diets containing vegetable proteins that are deficient in sulfur amino acids.


Population biology of Camallanus oxycephalus Ward and Magath, 1916 (Nematoda: Camallanidae) in white bass in western Lake Erie.

The distribution and abundance of the nematode Camallanus oxycephalus infecting white bass, Morone chrysops, in western Lake Erie was studied for over 2 years. Infection was generally more frequent and of higher intensity in large fish. The frequency distributions of nematode abundance in all segments of the fish population followed the negative binomial distribution. The data show seasonal cycles in population structure, site selection, intensity of infection, maturation, and reproduction. Infection occurs during July and August with a resulting peak in population density; during late summer and autumn, mortality, probably density-dependent, reduces the population by 30 to 60%; surviving worms are eliminated at 1 year of age. Growth and development of female worms is arrested from November to April, then proceeds at a rapid rate until the worms release their larvae and die. This growth pattern is probably related to temperature but may also involve host hormone cycles. The dispersal period of the nematode coincides with the annual maximum density of the intermediate host, a cyclopoid copepod,and is interpreted as an adaptation which increases the probability of successful transmission. Because the number of larvae produced by each female worm is a function of body volume, natural selection has favored rapid spring growth and attainment of large body size relative to the male worm. Both seasonal timing in the life cycle and the number of larvae produced are important factors in determining the abundance of this and perhaps other parasites. Evidence is presented suggesting that fluctuations of environmental parameters may disrupt the timing of transmission and alter the distribution and abundance of the parasite. It is hypothesized that the magnitude of such changes in parasite abundance may be related to the complexity of the host-parasite system.


Ultrastructure of cytoplasmic and nuclear changes in Eimeria tenella during first-generation schizogony in cell culture.

Eimeria tenella sporozoites were inoculated into primary cultures of chick kidney cells. Cells fixed from 1 1/2 to 54 hr later were examined with the electron microscope. At 1 1/2 and 24 hr, most intracellular sporozoites were fusiform and retained organelles typical of extracellular sporozoites. However, at 35 hr, rounded trophozoites were present without these structures; only a refractile body, nucleus, mitochondria, and endoplasmic reticulum remained. Binucleate parasites were also present at that time, but at 48 hr many multinucleate schizonts were present. Nuclei, with adjacent conoids, were at the periphery of these schizonts. Partly developed merozoites, each containing a conoid and a nucleus, protruded into the parasitophorous vacuole. At 54 hr, fully developed merozoites were separated from the residual body. Merozoites resembled sporozoites but lacked the large refractile bodies seen in sporozoites. Linear inclusions were present near the merozoite nucleus and in the residual body. Round vacuoles and ribosomes were also found in the residuum. Nucleoli were first seen in sporozoite nuclei at 1 1/2 hr. They were also present in merozoites but were more prominent in trophozoites and schizonts. Peripheral and scattered nuclear heterochromatins were prominent in intracellular sporozoites and diminished in trophozoites, but increased after several nuclear divisions and were again prominent in the merozoite. Small, distinct interchromatin granules were found in all stages. Intranuclear spindles, centrocones, and centrioles were found in connection with nuclear divisions. Ultrastructure of first-generation schizogony in cell culture was similar to that described for second-generation E. tenella in the chicken and to schizogony of other species of Eimeria.


The involvement of corticosteroids in the supersensitivity produced in the rat anococcygeus muscle by morphine withdrawal, thyroidectomy or a single dose of reserpine.

The effects of chronically administered corticosterone on the responses of the rat isolated anococcygeus muscle to agonists (acetylcholine, norepinephrine) were compared with those of morphine withdrawal in intact or metyrapone-pretreated rats and with those of a single dose of reserpine in intact or adrenalectomized rats.  The possibility that the similar supersensitivity, which occurs in thyroidectomized rats, might be associated with elevated corticosterone levels was investigated by measuring fluorimetrically the plasma levels of corticosterone in thyroidectomized rats.  Corticosterone, morphine withdrawal or a single dose of reserpine produced similar supersensitivities, characterized by increased maximum responses to both agonists (acetylcholine, norepinephrine) and an increased pD2 value for acetylcholine.  This effect of morphine withdrawal together with the weight loss characteristic of this condition was absent in metyrapone-pretreated rats.  A single dose of reserpine had no effect in adrenalectomized rats.  Plasma corticosterone levels were elevated in thyroidectomized rats. These results suggest that the common factor underlying the identical supersensitivities produced by corticosterone, morphine withdrawal, a single dose of reserpine or thyroidectomy is the ability of each treatment to raise plasma corticosterone levels.


Uterine response to ovariectomy during the proliferative and luteal phases in the marsupial, Trichosurus vulpecula.

The uterine luteal phase in T. vulpecula is not dependent upon the secretions of the CL throughout its duration. Ablation of the CL or ovariectomy after Day 7 of the 26-day oestrous cycle does not result in the termination of the uterine secretory phase. The dependence of the luteal phase on the secretions of the CL is demonstrated by ablation of the CL or ovariectomy on Days 2, 4, 8, 12 and 24 of the oestrous cycle. Ablation of the CL before Day 8 resulted in the inhibition of the impending luteal phase, and the commencement of a follicular phase resulting in oestrus 8 to 9 days later. Removal of the CL or ovariectomy on Days 8 or 12 does not completely inhibit the uterine luteal phase since sufficient precursor of uterine milk is stored in the uterine basal glandular epithelium, thus enabling the endometrium to maintain the secretion of uterine milk.


Changes in blood histamine and bradykininogen levels during different periods of the oestrous cycle in goats.

Blood histamine and plasma bradykininogen levels were estimated in eighteen normally cyclic goats at different periods of the oestrous cycle. The histamine level increased significantly (P smaller than 0.05) on Day 18 of the oestrous cycle. The bradykininogen level decreased significantly (P smaller than 0.05) towards the end of oestrus.


Isolation of a Sindbis virus variant by passage on mouse plasmacytoma cells.

A variant of Sindbis virus has been isolated by growing a stock of virus, previously passaged on chicken embryo cells, in mouse plasmacytoma (MOPC 315) cells in suspension culture. An indirect immunofluorescence test and infective center assay showed that only a small fraction of cells could be infected by the stock wild-type virus, but that the population of virus accumulating after a few passages on the mouse cells had host-range properties distinct from the stock virus. The mouse-passaged virus retained its virulence for the original host and back-passaging of this virus on chicken cells did not change its newly acquired properties. Thus, this variant appears to be a genetically distinct form of Sindbis that adsorbs to and grows much better than the stock virus on several types of mouse cells including cultures of mouse macrophages.


Protein metabolism during the steady state of Newcastle disease virus infection. I. Kinetics of amino acid and protein accumulation.

Pulse-chase experiments and studies on the effects of varying pulse lenghts on radioactive amino acid and protein accumulation have been carried out to evaluate several possible mechanisms for the inhibition in cellular protein accumulation during infection of chicken embryo cells by Newcastle disease virus. The inhibition is probably at the level of synthesis of cellular protein since no evidence for either increased degradation of protein or alterations in cellular permeability to protein was found in infected cultures. The magnitude of the reduction in the rate of cellular protein accumulation and consequently total protein accumulation depend upon the length of the radioisotopic labeling period. In contrast, the rate of viral protein accumulation is independent of the length of the labeling period. A double-label difference analysis of polyacrylamide gels was used in all of the kinetics studies to distinguish between viral and cellular protein accumulation. An unstable fraction which could be labeled with radioactive amino acids was detected in both infected and uninfected cultures. This material migrated mainly in the 50,000- to 60,000-dalton region of polyacrylamide gels, exhibited saturation kinetics during accumulation studies, and turned over rapidly during a chase. The relative amount of this fraction was not affected by infection. Gel analysis of the radioactive protein recovered from the medium from both infected and uninfected cultures revealed a major component with an apparent molecular weight of 33,000. None of the major viral polypeptides could be detected in the medium after a 30-min chase following a brief labeling period.


Structural proteins of two salmonid rhabdoviruses.

Purified infectious hematopoietic necrosis (IHN) virus and the virus of haemorrhagic septicaemia (VHS) (Egtved virus) each contain five structural proteins which were designated L, G, N, M-1, and M-2. The IHN viral polypeptides have molecular weights estimated to be 157,000, 72,000, 40,000, 25,000 and 20,000, respectively, whereas those of VHS viral polypeptides are estimated to be 157,000 74,000, 41,000, 21,500, and 19,000, respectively. The carbohydrate composition of the glycoprotein (G) was confirmed by demonstrating selective incorporation of [3H]glucosamine into the designated G protein of both viruses. Phosphoproteins were identified by incorporation of [32P]orthophosphate into the N and M-1 proteins of IHN virus and into the N protein of VHS virus. The glycoprotein of each virus was selectively solubilized by treatment with Triton X-100 in low salt buffer, whereas the M-1, and M-2 proteins along with the G protein were solubilized by Ttition X-100 in 0.43 M NaCl. The protein composition of the salmonid rhabdoviruses resembles that of the rabies virus group more closely than the vesicular stomatitis virus group.


Differential accumulation of virus-specific RNA during the cell cycle of adenovirus-transformed rat embyro cells.

In adenovirus type 2-transformed rat embryo cells there is a threefold greater incorporation of [3-H]uridine into virus-specific RNA early in S phase than in late S or G2. This heightened accumulation of labeled RNA is true for both nuclear and cytoplasmic virus-specific labeling. Inhibition of DNA synthesis decreases the virus-specific RNA labeling, whereas reversal of inhibition again allows the elevated level of virus-specific RNA labeling.


Vascular patterns of testicular tumors: a microangiographic study.

A microangiographic study of 12 cases of testicular tumors was done. In seminoma the blood supply was meager and centripetal, and embryonal carcinoma of adult type was hypervascular with coiling vessels. Vascular patterns of embryonal carcinoma of infants were sinusoidal. Choriocarcoma was hypervascular and there were multiple defects within vessels by its invasion. In a teratomatous cyst vessels of the wall were sometimes irregular, as indicated by surviving teratomatous elements, while in an epidermoid cyst fine vessels of the wall ran parallel in a regular manner.


The heritability of blood pressure: an investigation of 200 pairs of twins using the cold pressor test.

Blood pressures in 200 pairs of twins having a mean age of 14.0 (S.D. equals 6.5) years were measured at basal levels and in response to the cold pressor test. Intraclass correlations were consistently larger (and intrapair variances consistently smaller) in monozygotic as compared to dizygotic pairs, substantially more so in females than in males. Sex differences in heritability were somewhate greater for systolic than for diastolic blood pressure. Indices of heritability (H) at basal levels were, in descending order: 0.78 (female systolic), 0.61 (female diastolic), 0.56 (male diastolic), and 0.41 (male systolic). The same descending order for H was found for blood pressure responses to the cold pressor test. It is concluded that in the population studied, genetic factors play an important role in the control of blood pressure over the normal range of vascular reactivity. In females, such factors may be the major determinant of the variability observed.


[Dynamics of carbohydrate metabolism in the heart muscle in early ontogenesis].

The dynamics of glycolysis and glycogenolysis in the postnatal period, determined from the accumulation of lactate in the homogenate of the heart muscle, indicates that the glycolytic activity begins to subside right after the birth and becomes constant after a lapse of 15-20 days. The activity of the phosphorylase, phosphohexoisomerase, enolase and pyruvate-kinase in the heart muscle extract was found to remain virtually unchanged. Some decline is noted in the activity of the aldolase and hexokinase, while that of the phosphofructokinase and lactate-dehydrogenase rises by 50-60 per cent. These data suggest that in the early ontogenesis the increasing capacity of the mitochondrial system in the heart muscle is paralled by an adjustable conditioned inhibition of the glycolytic phosphorylation, with concurrently rising activity of the phosphofructokinas and, consequently, also of the potential capacity of glycolysis and glycogenolysis.


Decomposition of unsaturated fatty acid hydroperoxides by hemoglobin: Structures of major products of 13L-hydroperoxy-9,11-octadecadienoic acid.

13L-hydroperoxy-9, 11-octadecadienoic acid was decomposed rapidly in the presence of hemoglobin. The product consisted of five major compounds, i.e. 13-keto-9, 11-octadecadienoic acid, 13L-hydroxy-9, 11-octadecadienoic acid, erythro-11-hydroxy-12, 13-epoxy-9-octadecenoic acid, threo-11-hydroxy-12, 13-epoxy-9-octadecenoic acid, and 9 DL-hydroxy-12, 13-epoxy-10-octadecenoic acid.


Calcitonin: a general survey.

Since the initial discovery of calcitonin, a considerable amount of information has been obtained concerning structure, function, secretion, and effects of the hormone from a number of species. The purpose of the present report is to review the studies of structure and function, the factors that influence secretion and synthesis, and the effects and mechanism of action of calcitonin. The clinical uses of the hormone and syndromes of calcitonin excess are discu,sed.


[Diagnostic and Therapeutical problems of extrahepatic portal vein stenosis].

In seven children the clinical course of extrahepatic portal vein stenosis was studied angiographically. In all children with splenomegaly it was deemed necessary to perform an X-ray barium examination of the oesophagus. Transsplenic portal venography was performed when oesophageal varices were detected to set up the best scheme for emergency treatment. In our patients with hypersplenism there were no specific bone marrow alterations. In contrast to the literature most of our cases had fibrosis of the liver.


[Diseases caused by drug therapy in newborn babies].

A drug may reach the newborn indirectly from the mother via the umbilical cord or brest milk and by direct application. If the Apgar score after 5 or 10 min decreases in relation to the score of the 1st minute, a depression by drugs has to be considered if other causes are eliminated. Some drugs and typical symptoms are described as for instance diazepam, addiction causing drugs, reserpin and imidazol derivatives. With regard to the direct effect of drugs the danger of the simultaneous application of several remedia is especially pointed out.


DiC-like syndrome after envenomation by the snake, Crotalus horridus horridus.

After envenomation by the timber rattlesnake (Crotalus horridus horridus) a young man showed massive swelling and ecchymoses of the involved extremity, generalized petechiae and a large hematoma of the left upper eyelid. Two weeks later he was completely well. The first blood sample was incoagulable and showed high titers of fibrin split products by the MISFI and staphylococcal clumping tests. Immounoelectrophoresis showed both D and E fragments, but only D was present at 18 hours. The profound thrombocytopenia and hypofibrinogenemia gradually improved, and split products disappeared. In vitro the venom had thrombin-like activity, clotting fibrinogen or plasma and aggregating platelets. These activities could be inhibited by antivenom but not by heparin. There was no evidence that thrombin formation was actually involved: hence, the syndrome is called DIC-like.


Twenty-four-hour secretory patterns of gonadotropins and prolactin in a case of Chiari-Frommel syndrome.

Plasma LH, FSH and prolactin secretory patterns were derived from the measurement of 20-min interval plasma samples obtained during a complete 24-h period in a patient with persistent postpartum amenorrhea and galactorrhea (Chiari-Frommel syndrome), before and after clomiphene citrate therapy. During nocturnal sleep, polygraphic monitoring was carried out to precisely identify sleep onset, specific sleep stages and waking periods. During the evening and nighttime hours, LH and FSH concentrations were markedly reduced, compared to the daytime patterns both before and after clomiphene therapy. A sleep associated rise of prolactin concentration was present, similar to the pattern found in normal subjects but at higher concentrations. The reciprocal nature of the nocturnal secretory patterns for LH and FSH and prolactin in this patient suggests an alteration in hypothalamic dopaminergic mechanisms which are thougt to control the secretion of these hormones.


Enhancement of the electrical excitability of neuroblastoma cells by valinomycin.

Mouse neuroblastoma cells in stationary phase of growth display partially developed electrical properties. Addition of the K+ selective carrier valinomycin to these cells causes rapid enhancement of electrical excitability. We suggest that the appearance of molecules with properties similar to valinomycin is essential for the full expression of electrical excitability in differentiating neuroblastoma.


[Pancreatic cysts].

True and pseudo-cysts of the pancreas are described and their aetiology, pathology, laboratory tests, radiological examination, differential diagnosis, symptomatology and surgical management are illustrated. A series of 22 cases of pancreatic cyst is presented. Surgical management consisted of 14 cystogastrostomies, 3 cystoduodenostomies, 2 resections of the tail of the pancreas, 1 internal drainage between the fistular segment of the gland and the gastric cavity, and 2 external drainages with a Pezzer tube. It is felt that internal drainage is the operation of choice. Of the surgical techniques available, a preference is expressed for cystogastrostomy and cystoduodenostomy.


A diagnostic device for stress incontinence.

Many methods of evaluating anatomic deformity of the urethrovesical angle are available. A new diagnostic device has been developed to determine the degree of rotation of the urethral axis, thus differentiating between type I and type II deformities causing stress incontinence. Preliminary study in 62 patients indicates that the instrument is accurate, simple, and easily interpreted. The device also has the advantage of being easily constructed, disposable, and inexpensive.


Symposium on sensorineural hearing loss in children: early detection and intervention. Evaluation of the child with sensorineural hearing impairment.

When the physician is confronted with a child with sensorineural hearing impairment, he should do everything in his power to establish the cause of the hearing loss in order to diagnose those conditions that might be corrected or controlled and to counsel the parent concerning the child's future. Certain physical characteristic should alert the physician to the fact that there may be an accompanying sensorineural hearing loss. The family, gestational, and postnatal history may help establish this diagnosis. Finally, laboratory and x-ray examinations should be performed in an attempt to arrive at a definitive diagnosis.


Nippostrongylus brasiliensis infections in mice: the immunological basis of worm expulsion.

Nippostrongylus brasiliensis infections in mice were terminated more rapidly than in rats and immunologically induced damage occurred earlier. Like rats, mice expelled damaged worms more rapidly than normal worms. Recipients of cells from the spleen or mesenteric node of immune mice expelled their worms by day 8 of the infection. Recipients of cells alone or antiserum alone did not expel their worms by day 5 but mice given both cells and antiserum expelled their worms by this stage of the infection. Damaged worms were expelled more rapidly than normal worms from mice given immune cells, This work indicated that antibodies and cells collaborate to expel N. brasiliensis from mice as has been shown to occur in rats.


Development of paw preference in rats following unilateral cortical ablations in infancy.

Rats which received unilateral anterior cortical lesions in infancy manifested a preference for the paw contralateral to the intact hemisphere when tested at 5 wk. and at 10 wk. of age. That this effect was locus specific and was not due to a change in the over-all balance of activity between the hemispheres was indicated by the fact that similar lesions made in the posterior cortex did not influence which paw the animals subsequently preferred. The posterior lesions, however, may have lowered the frequency of ambidexterity, a finding interpreted in terms of theoretical arguments concerning bilateral symmetry and the discrimination and memory of left and right.


Handedness as a function of test complexity.

Handedness as a function of test complexity was investigated with two tests, Dotting and Tapping-on-squares. For each one three levels of complexity were established by varying radius (Dotting) and square sides (Tapping-on-squares). Administration of the different complexity levels to a total of 310 third and fourth grade boys and girls showed that the rate of left-handedness increased from 6% to 16% with decreasing level of test complexity. It is concluded that optimal classification of handedness is only made possible by maximal levels of test complexity, which have to be established empirically for different populations.


Electrical activity in embryonic heart cell aggregates. Developmental aspects.

Action potential parameters were measured in beating heart cell aggregates which were formed from trypsin-dissociated cells of embryonic chick heats aged 2 1/2, 4 or 7 days. 1. In aggregates composed of cells from the whole heart there was an increase in the maximum diastolic potential, overshoot, maximum rate of rise of the action potential (V max), and action potential duration between days 2 1/2 and 7. 2. Action potential parameters from 4- or 7-day aggregates composed exclusively of atrial or ventricular cells were similar to those in whole heart aggregates of the same age with the exception of the action potential duration in which atrial less than whole heart less than ventricular. Between days 4 and 7 the increases in duration were approximately 14% in atrial, 35% in whole heart, and 50% in ventricular aggregates. Differences in action potential duration, within or between ages, were not due solely to differences in the rate of beating. 3. Action potentials in whole heart aggregates aged 2 1/2 days were insensivitive to TTX (10-5 g/ml) but abolished by D600 (1 MUG/ML). Conversely, at 7 days activity was suppressed by TTX (2 X 10-8 G/ML) WHILE D600 (1 mug/ml) shortened the action potential duration and reduced the overshoot without influencing V max. 4. Adrenaline (1 mug/ml) restored the action potential overshoot and duration in 7-day aggregates treated with D600. 5. Action potential development in embryonic heart cells appears to be characterized by the functional appearance of fast inward channels. The slow channel mechanism, previously utilized in action potential generation, may gradually assume its adult role of carrying inward current during the plateau phase. 6. In contrast to monolayer cultures, embryonic heart cells cultured in aggregate form seem to have membrane properties similar to those of intact tissue.


Diseases of the breast.

As a result of health education programs that have stressed self-examination and the importance of early diagnosis and treatment, less than 5 percent of breast lesions seen in clinical practice today are malignant. Nevertheless, breast cancer is the leading cause of deaths due to cancer in women and the second most common breast lesion. Chronic cystic mastitis, the most common breast lesion seen in women, can often be treated conservatively by aspirating the cystic fluid. Close follow-up is necessary so that if carcinoma develops, it can be treated early. Carcinoma is unpredictable but not hopeless. Although controversy exists regarding treatment, radical mastectomy is still the preferred treatment in most cases. Other breast diseases include adenofibroma, a common lesion requiring excisional biopsy for positive diagnosis; nipple discharge, which should be carefully examined cytologically; and relatively rare breast lesions, including intraductal papilloma, adenosis, traumatic fat necrosis, and cystosarcoma phylloides.


The radiosensitization of hypoxic tumor cells by hyperthermia.

The influence of hyperthermia on the survival of irradiated oxic and hypoxic HeLa cells was determined. A significant reduction in OER was obtained for irradiated cells subsequently cultivated at 42-degrees C for two hours compared to irradiated cells cultivated at 37-degrees C. These results suggest that heating irradiated hypoxic cells is an effective means of reducing the oxygen dependence factor in radiation effects.