Purification of interferon from mouse Ehrlich ascites tumor cells. Interferon production was induced in mouse Ehrlich ascites tumor cells by infection with Newcastle disease virus. The interferon produced was purified by precipitation with ammonium sulfate, chromatography on carboxymethyl-Sephadex, treatment with blue dextran and polyethylene glycol, gel filtration on Bio-Gel P-60 and Bio-Gel P-200, chromatography on phosphocellulose, isoelectric focusing, and chromatography on octyl-Sepharose. The specific activity of the product was 1.6 x 10(9) NIH mouse interferon reference standard units/mg of protein. Electrophoresis in polyacrylamide gels in the presence of sodium dodecyl sulfate indicated that the apparent molecular weight of the interferon-active material ranged from 25,000 to 35,000. As revealed by staining the gels with Coomassie brilliant blue, the interferon activity co-migrated with the major, broad protein band. Minor, stainable bands of proteins were free of interferon activity and their apparent molecular weight was smaller than 12,000.


Newly synthesized acetylcholine receptors are located in the Golgi apparatus. Chick skeletal muscle cells in tissue culture were fixed and treated with saponin to allow [125I]alpha-bungarotoxin access into the cells while preserving ultrastructure. The kinetics of binding of iodinated alpha-bungarotoxin to intracellular acetylcholine (ACh) receptors and to surface A Ch receptors were comparable. About half of the intracellular ACh receptors are newly synthesized and in the pathway leading to incorporation into the plasma membrane. Correlated electron microscope autoradiographic and kinetic studies of this receptor population suggest that a substantial fraction of the newly synthesized ACh receptors are located in the Golgi apparatus, where they reside for approx. 2 h.


Sythesis of tropomyosin in cultures of differentiating muscle cells. The accumulation of tropomyosin in cultures of differentiating muscle cells was quantitatively measured. Tropomyosin was isolated from cultured cells during and after myoblast fusion; both alpha- and beta-subunits were present in myotube cultures. During fusion small amounts of tropomyosin were detectable, but, as fusion approached a maximum, tropomyosin accumulation began to increase. The increased synthesis of tropomyosin after the initiation of muscle cell fusion is consistent with the increased synthesis of other proteins characteristic of muscle, including myosin.


The mechanism of killing of mouse fibroblasts by the amino acid analogue 5-fluorotryptophan. The mechanism of killing of A9 fibroblasts by 5-fluorotryptophan has been studied. L-tryptophan competitively relieves the growth inhibition caused by 5FT. After incubation with 5FT, 3H-5FT was incorporated into protein, replacing tryptophan residues. During the initial hours of incubation with 5FT, a specific inhibition was observed of the incorporation of L-tryptophan into protein; later this inhibition was followed by a general inhibition of protein synthesis and cell division. However, nuclear division continued after cell division had ceased. While 5FT was observed to be incorporated into protein after a 1 hour period in MEM + 0.40 mM 5FT in A9, no 3H-5FT was incorporated into protein in a mutant isolated by its resistance to killingy by 5FT. These results support the hypothesis that cell death occurs due to malfunctioning proteins which contain 5FT residues.


Expression of nervous system antigen-3 by lines of mouse fibroblasts and kidney cells and continued expression in hybrid cells. In a survey of the expression on cultured mouse cells of the cell surface antigen known as nervous system antigen-3 (NS-3), it was found that RAG, a renal adenocarcinoma line, expressed that antigen. It was also observed that 3T3, a fibroblast line of unknown tissue origin, expressed NS-3. Cells of these two lines were hybridized with cells of two mouse L cell lines that did not express NS-3. Four hybrid clones were tested for both the 3T3 X L cell cross and the RAG X L cell cross, and all the hybrids were found to be NS-3 positive. All the hybrids had at least 40% as much activity as the NS-3 positive parent. Of the four parental mouse cell lines used, only 3T3 expressed Thy-1.2 antigen on the cell surface. In contrast to the continued expression of NS-3 on hybrid cells, Thy-1.2 antigen was not detectable on two clones of 3T3 X L cell hybrids that were tested.


The regulation by fibroblast growth factor of early transport changes in quiescent 3T3 cells. This study involves the use of fibroblast growth factor (FGF) as a substitute for exogenous serum to examine the early transport changes which occur when quiescent 3T3 cells re-initiate active growth. FGF, in nanogram amounts, together with insulin and dexamethasone, can induce mitogenesis and mitosis in 3T3 cells GO-arrested by holding in growth medium containing 0.8% calf serum. In terms of quiescent cell transport activity enhancement, FGF is 300,000-fold more effective than fresh serum, on a protein basis. In addition, very short exposure of serum-depleted cells to FGF indicates that a distinct temporal or time sequence exists in the transport system activation process. For example, uptake of alpha-aminoisobutyric acid (AIB) and uridine are stimulated very rapidly, whereas hypoxanthine uptake does not respond until much later. Closer analysis shows that AIB uptake is maximally enhanced within zero to two minutes after FGF addition to cells. Finally, the stimulatory effect of FGF on transport system activities is specific in terms of the proliferative state of the cells to which it is added, and in terms of the uptake systems which respond to it.


Reversible inhibition of the hydrocortisone induction of glycerol phosphate dehydrogenase by cytochalasin B in rat glial C6 cells. The hydrocortisone (HC) induction of glycerol phosphate dehydrogenase (GPDH; EC 1.1.1.8) in rat glial C6 cells was inhibited reversibly and in a dose-dependent manner by cytochalasin B (CB). CB had no effect on basal level GPDH, total cellular RNA, DNA or protein content nor did it act as a general inhibitor of the rate of protein synthesis. CB did not appear to be acting via dissociation of microtubules since colcemid had no effect on the induction process. The addition of an alternate energy source (sodium pyruvate) did not relieve the CB inhibition of GPDH induction suggesting that CB is not exerting its effect by blocking glucose utilization. The inhibition by CB is not dependent on the temporal sequence of the induction process since it specifically inhibited GPDH induction at any time it was added. CB did not alter the rate of degradation of GPDH in these cells and direct measurements of the specific rate of synthesis of GPDH demonstrated that CB decreased the induced rate of GPDH synthesis by about 60%. The site of inhibition was more precisely defined  by experiments which demonstrated a 60% decrease in specific nuclear binding of 3H-HC even though total cellular uptake of 3H-HC was unaffected. This effect on nuclear binding of HC is sufficient to account for the decreased accumulation of GPDH activity in CB-treated cells.


Assessment of incompetence in canine aortic and pulmonary valve grafts and comparison of some methods of valve preservation. A new method has been devised to assess the competence of the canine aortic valves, before implantation. This method would provide a better selection of aortic valves before their insertion and may reduce the incidence of postoperative incompetency. Comparison of canine pulmonary and aortic valve function, at various pressures, revealed that the pulmonary valves were regurgitant at higher pressures, in comparison to aortic valves. Formalin preserved valves were found to be significantly regurgitant.


Polyamine metabolism in enucleated mouse L-cells. The distribution of polyamines between the nucleus and the cytoplasm, and the role of the nucleus in polyamine metabolism, have been studied using cells enucleated with cytochalasin B. Spermidine and spermine were found in the nuclear and the cytoplasmic fractions of L929 cells; their concentration was 3-fold higher in the former fraction. Ornithine decarboxylase activity was only found in the cytoplasm, and this activity could be stimulated in enucleated cells by the addition of fresh medium. These cells synthesized putrescine actively, but the putrescine made was not converted to spermidine, and accumulated to relatively high concentrations. Similarly, methionine did not act as a precursor to spermidine in enucleated cells, in contrast to whole cells, although it was incorporated into cell protein. Spermidine synthesis, unlike putrescine synthesis, appears to be completely dependent on a nuclear component.


On the mechanism of the Crabtree effect in mouse ascites tumor cells. The addition of glucose to ELD and ELT/B1 mouse ascites tumor cell suspensions caused a 2.3-fold increase in the phosphorylation state ratio, (ATP)/(ADP) (Pi), because of a decrease in the intracellular Pi concentration. The addition of glucose to these cell suspensions has been reported by Chance and Hess ('59) to cause an increase in the study state reduction of cytochrome b and an increase in the steady state oxidation of cytochrome c. On a quantitative basis these two independent measurements suggest that a near equilibrium exists between the oxidation-reduction state of the mitochondrial electron carriers and the reactions of ATP synthesis (as expressed by the phosphorylation state ratio) both before and after glucose addition. We conclude that the mechanism of the inhibition of respiration by glycolysis (the Crabtree effect) is a decrease in the rate of electron transport caused by the mass action effect of the elevated phosphorylation state ratio.


Growth stimulating activity in bovine pituitary extract specific for a rat mammary carcinoma cell line. We report the isolation of a bovine pituitary growth factor (MGF) for a rat mammary carcinoma cell line, 64-24, which was isolated from a highly hormone-dependent mammary tumor. The MGF has been partially purified by a series of Diaflo ultrafiltration membrane sievings, isoelectric focusing and Sephadex columns. The MGF has a molecular weight of approximately 1,000 to 2,000 daltons and has a U.V. absorption spectrum typical for a polypeptide. Its isoelectric point is approximately pH 3.8-4.0. The factor is heat stable. The growth stimulating activity of the MGF does not stimulate other rat cell lines (22-1, RMG or HTC lines) but is specific for the 64-24 mammary tumor cell line. The MGF is not among previously reported pituitary hormones or growth factors.


ATP, trehalose, glucose and ammonium ion localization in the two cell types of Dictyostelium discoideum. Ultra-microfluorometric techniques were adapted to follow several compounds related to energy metabolism through the developmental cycle of Dictyostelium discoideum. Each compound (ATP, trehalose, glucose, and ammonium ion) was found to be present in stalk and/or spore cells. The accumulation of NH4+ was interpreted as an indication of protein degradation, a source of energy in this organism. During the early stages of differentiation NH4+ was localized only in prestalk cells. However, it accumulated in spore cells during culmination such that levels were comparable in the two cell types by the end of development. Trehalose, an energy source for germinating spores, was found in both cell types but was preferentially degraded in stalk cells late in development. Glucose, the degradation product of trehalose, was localized in prestalk cells and varied inversely with trehalose levels. ATP was not localized in a specific cell type during development. However, ATP declined in stalk cells at an earlier stage of development.


Continuous DNA replication in the nucleus of the dinoflagellate Prorocentrum micans (Ehrenberg). The uptake of tritiated thymine into cells of a heterogeneous population of Prorocentrum micans was investigated using light-microscope and electron-microscope autoradiography. Specificity of thymine uptake into DNA was demonstrated by the specific removal of label from wax-embedded material using DNase and by the high degree of localization of nuclear label to chromosomes in the electron-microscope autoradiographs. All nuclei, including both dividing and non-dividing cells, showed a substantial uptake of label, indicating that nuclear DNA synthesis in Prorocentrum micans is a continuous process. The level of DNA synthesis does show considerable variation, however, with very high levels in some interphase nuclei. The continuous replication of nuclear DNA provides further evidence of dinoflagellate affinity to the prokaryotes, and indicates that Prorocentrum micans is a very primitive eukaryote cell.


Feulgen microdensitometry and analysis of S-phase cells in cervical tumour biopsies. Tissue specimens from the cervical tumours of 70 patients undergoing radiotherapy were examined by Feulgen microdensitometry. Twenty-five of the 70 specimens were also subjected to in-vitro tritiated thymidine autoradiography to determine the proportion of DNA synthesising cells they contained. A spectrum of frequency distributions of nuclear DNA content was obtained from Feulgen microdensitometry, but by inspecting the data the basic DNA content of malignant cells could be established in most cases. Fifty-nine per cent of the tumours were 'diploid', 10% 'tetraploid', 13% 'diploid to tetraploid', and the remaining 18% 'aneuploid'. Grafical analysis of DNA frequencies from 48 'diploid' or 'tetraploid' tumours enabled the proportion of DNA synthesising (S) cells to be estimated by frequency distribution analysis. Estimates of the S component ranged from nil to 30%, were log normally distributed, and comparable to direct measurements of cells in S determined by autoradiography for 25 cases (range 1.2-28.7%). For all paired data the mean difference was 1.2 +/- 1.45% (confidence limits), suggesting that overall Feulgen microdensitometry analysis may be an equally valid technique in providing cellular kinetic information with human tumour material.


Ultrastructure of dyskeratosis in morbus Darier. Biospy specimens from three patients with morbus Darier were studied by electron microscopy: Grains appeared in groups in the midst of dyskeratotic lesions and were recognized as being the result of more advanced acantholysis and precocious keratinization. Corps ronds were formed individually in the regions lateral to that of grains, where hypergranulosis was prominent in contrast to a mild dyskeratosis. Therefore, the formation of grains and corps ronds seem to be independent of each other. While in the literature acantholysis has mainly been ascribed to the separat;on of desmosones into two segments, another initial process seems to be the primary disappearance of the attachment plaque along with the connecting tonofilaments. From the existence of the other contact complexes, it seems likely that a restoration of desmosomes between acantholytic cells should take place. In early stages of dyskeratotic cells, keratinosomes were increased in number and some of them persisted inside the plasma membrane without a release into the intercellular spaces, and thus they were present in grains and corps ronds. The dyskeratosis in morbus Darier is characterized by a premature, incomplete and individual keratinization as evidenced by the existence of keratohyalin granules and keratinosomes as early as at the level of the 3rd epidermal layer and the absence of a keratin pattern in the dyskeratotic horny cells.


Demonstration of antibodies to soluble nuclear antigens by complement fixation test. An application of microtiter technique. The complement fixation test utilizing the microtiter system was applied to study sera from patients with a variety of connective tissue diseases for the presence of antibodies to soluble nuclear antigens. A good correlation was found between microtiter complement fixation and hemagglutination for anti-RNP and anti-Sm antibodies. Anti-Ha antibodies were detected by complement fixation but not by hemagglutination. Different enzyme digestions of the soluble nuclear antigen prior to assay and use of a quantitative microcomplement fixation test were found to improve the analysis of sera with multiple antibodies.


Effect of oestradiol infusion on plasma gonadotrophins and ovarian activity in progesterone-primed and unprimed anoestrous ewes. Plasma gonadotrophin and ovarian responses to physiological infusions of oestradiol were examined in anoestrous ewes with and without progesterone priming. Intravenous infusion of oestradiol (1 microgram/h) for 24 h followed at 96 h by a second infusion for 12 h (low dose) resulted in simultaneous increases in LH and FSH of magnitude and duration similar to those of ewes at natural oestrus. Infusions of 3 microgram/h for 16 h, and again at 96 h for 4 h (high dose) resulted in similar gonadotrophin peaks. Endoscopy revealed that only two out of eight ewes had ovulated despite increased ovarian activity and surges of plasma gonadotrophins. Other anoestrous ewes were primed with 25 mg progesterone daily for 12.5 days before similar infusions of oestradiol on day 15. In the presence of very low plasma progesterone concentrations (less than 1.0 ng/ml) at the onset of infusion, there was no pituitary response to the infusion of a low dose of oestradiol but the gonadotrophins were released after the second high-dose infusion. Progesterone inhibited the positive feedback on both gonadotrophins but not the negative feedback on FSH. Ovulation occurred in all four ewes which had been progesterone-primed and infused with a high dose of oestradiol.


Protein synthesis in the early Drosophila embryo; analysis of the protein species synthesized. The soluble proteins were extracted from Drosophila eggs which had been permeabilitzed and incubated in medium containing [35S]methionine. These proteins were analysed on immunoelectrophoresis plates and on SDS polyarcylamide gels both by staining for total protein and by autoradiography. The radioactive proteins must have been synthesized during the period of incubation with [35S]methionine. In the period covered by this study (O-3 h) there was much protein synthesis but no new proteins were synthesized which had not already been synthesized during oogenesis. We conclude that the considerable protein synthesis that occurs in early Drosophila development is translated from maternal mRNA which is activated both by egg deposition and fertilization. Translation of protein from either masked maternal mRNA, which had not been previously translated, or from mRNA transcribed from the zygote genome must occur after blastoderm formation.


The culture of mouse blastocysts in the presence of uterine flushings collected during normal pregnancy, delayed implantation and pro-oestrus. When day-3 mouse embryos were cultured in a simple medium supplemented with uterine fluids of mice autopsied on day 4 of pregnancy, 48 h after administration of oestradiol, or during pro-oestrus, the percentage of blastocysts hatching from the zona pellucida was significantly greater than in unsupplemented medium. In the presence of uterine fluids recovered during delayed implantation this stimulation of blastocyst hatching was not observed. When the culture medium was supplemented with dialysed uterine flushings containing 20 or 30 microgram protein/ml, both 'day 4' and 'delay' uterine proteins were equally effective in enhancing hatching frequency (P less than 0.05). The results suggested that 'delay' uterine fluids may contain a dialysable inhibitor of blastocyst activity. The putative inhibitor was not effective in the presence of serum, since uterine fluids recovered both on day 4 of pregnancy and during delayed implantation significantly increased the size attained by blastocyst outgrowths in the presence of foetal calf serum (P less than 0.01). The percentage of blastocysts exhibitng giant cell transformation and outgrowth was also increased (P less than 0.02) by these uterine fluids when the concentration of FCS in the medium was minimal (0.25%).


Report of a case of true hermaphroditism with karyotype 45,X/46,XX/46,X,mar/47,XX,mar. The case of a 24-year-old man with hypoplastic external genitalia, lack of the right scrotal testis and gynaecomastia has been described. In the intermitotic cells the cytogenetic investigations revealed the presence of the X body and the absence of the Y body. A 45,X/46,XX/46,X,mar/47,XX,mar karyotype could be established. On laparotomy a rudimentary ovary, uterus and vagina were detected on the right side of the abdominal cavity.


Serum vitamin A in premature and term neonates. Serum vitamin A was determined in premature and term neonates by a specific spectrofluorometric method. Premature neonates (N = 42; gestational age = 32 +/- 0.4 weeks) had a serum vitamin A level (14.9 +/- 0.98 microgram/dl) significantly lower (P less than 0.001) than that of term neonates (N = 51; 22.4 +/- 0.99 microgram/dl). The vitamin A mean serum values of infants of 36 weeks' gestational age were not statistically different from those of the term neonates. Linear regression analysis for serum vitamin A values vs gestational age showed no significant correlation. A linear correlation (P less than 0.05), however, was found between serum vitamin A and serum protein protein concentrations, perhaps indicative of a lower concentration of retinol-binding protein. Since vitamin A is involved in the promotion of mucous-secreting cells, the premature neonate may be at greater risk than the term infant for diseases involving the mucosal epithelium, including necrotizing enterocolitis.


When I was born: perceived parental reactions of adolescents. A 32-item multiple choice test, When I Was Born, was designed to measure adolescents' perceptions of parental reactions at the time of their births. A total of 372 subjects, ages 11 through 20 years, were administered the test. The population included 140 physically normal subjects, 97 adolescents with acquired physical problems, and 135 subjects with congenital physical anomalies. Factor analyses of test data using 100 normal subjects as the criterion population identified four factors: Parental Emotion, Parental Apprehension, Parental Pride, and Parental Nurturance. On all four factors, normal subjects had the highest scores, and subjects with congenital anomalies had the lowest scores. The findings suggest that the test measures both adolescents' projected feelings about parental reactions at their birth and the current impact of a physical problem on the child.


New in vivo evidence for narcotic agonistic property of leucine-enkephalin. Administration of leucine-enkephalin or morphine to mice rendered dependent on morphine by pellet implantation inhibited the naloxone-precipitated abstinence syndrome. The withdrawal jumping response was inhibited by morphine or leucine-enkephalin; however, both failed to inhibit withdrawal defecation and rearing behavior. On a molar basis, leucine-enkephalin was half as potent as morphine in inhibiting the abstinence syndrome. New in vivo pharmacological evidence for narcotic agonist-like activity of leucine-enkephalin is provided.


Chemical constituents of gentianaceae XXIII: tetraoxygenated and pentaoxygenated xanthones and xanthone O-glucosides of Swertia angustifolia Buch.-Ham. The whole plant extract of Swertia angustifolia Buch.-Ham., collected at different stages of growth, contained 14 tetraoxygenated and five pentaoxygenated xanthones and xanthone 1-O-glucosides. Of the eight xanthone 1-O-glucosides isolated, five were previously unreported in nature. The xanthones are broadly based on 1,3,5,8- and 1,3,7,8-oxygenated systems, with an added oxygen function at C-4 in some compounds, and represent a number of methoxylated patterns. The content and relative abundance of the free xanthones and their 1-O-glucosides changed with plant growth. These results are the first demonstration of the variation in chemical characters in the different parts of a Swertia species during its ontogeny. The biolgocial significance of these results is appraised.


Three-dimensional molecular illustrations I: Isoelectron density contours and isoelectrostatic energy contours. A method of depicting dimensional illustrations of molecules in vacuo that are sensitive to small electronic perturbations was attempted. This method would be useful in determining the effects of either perturbing groups from other molecules or changes produced by the addition or modification of an existing atom or chemical group on the same molecule. Isoelectron density contours for small molecules such as benzene, ammonia, and formaldehyde were first considered using the CNDO/2 molecular approximation method and then extended to the use of deorthogonalized CNDO/2 eigenvectors. These methods were similar in molecular projections but insensitive to electronic alterations. Therefore, the electrostatic potential energy was considered in developing contour surfaces of several of the molecules studied. In this case, acute and visually discernible changes were evidenced by electron exchange in the three-dimensional illustration of formaldehyde. The effect on the two-dimensional contour map of ammonia was strikingly altered by the addition of a proton, further substantiating the sensitivity of electrostatic contours to perturbing influences. These methods are considered and amplified in this report.


Potential hypocholesteremic derivatives of styrylacetic acid II: cis- and trans-3-methyl-4-phenyl-3-butenoic acid analogs. The synthesis and preliminary biological testing for in vitro cholesterol biosynthesis inhibitory activity of 2-indeneacetic acid, 2-methyl-1,2-dihydro-2-naphthoic acid, and their 5- and 7-chloro derivatives, respectively are described. These compounds were prepared as trans- and cis-analogs of the known antilipemic agent 3-methyl-4-phenyl-3-butenoic acid. Although both series of compounds showed cholesterol biosynthesis inhibitory properties, chloro substitution enhanced potency only in the cis-system. These findings are discussed in terms of a possible relationship between the cis-compounds and clofibrate-type antilipemic agents.


Effect of general anesthetics on handling- and decapitation-induced increases in sympathoadrenal discharge. The effect of five anesthetics--cyclopropane, pentobarbital, urethane, chloralose or ketamine hydrochloride--on handling- or decapitation-induced increases in adrenergic tone were studied in the intact rat. The anesthetic agents tested prevented or markedly reduced stress-induced increases in levels of plasma total catecholamines and norepinephrine. Similar changes in norepinephrine were seen in corticosterone- treated adrenalectomized rats in which this catecholamine seemed to be the only one present in plasma. During anesthesia with cyclopropane, blood pressure fell; there was no additional decrease in total plasma catecholamines when the concentration of the inhaled anesthetic agent was increased. With increased length of cyclopropane anesthesia, however, total catecholamine and norepinephrine concentrations increased. Thus, the different effects of anesthetics on the cardiovascular system cannot be solely explained by their effects on stress-induced increases in sympathetic neuronal activity as reflected by circulating catecholamine levels.


Effect of cold exposure on mammary circulation oxygen consumption and milk secretion in the goat. 1. Measurements of milk secretion, mammary oxygen consumption and blood flow, cardiac output and blood pressure have been made in lactating goats exposed to a thermoneutral environment and to two levels of cold stress which were sufficient to raise total body oxygen consumption by an average of 18 and 46% above that in a thermoneutral environment. 2. Mammary blood flow and oxygen consumption were not appreciably affected by the milder cold stress but fell to 63 and 70%, respectively, of their thermoneutral value in the moderately cold environment. Resistance to blood flow through the udder was increased by this level of cold stress and the gland took a smaller fraction of an increased cardiac output. 3. Both levels of cold exposure caused a reduction in milk secretion, to 80 and 40% of its thermoneutral value in the mild and moderate cold respectively. The amount of milk secreted per unit volume of blood flowing through the gland decreased as the severity of the cold stress increased. 4. Voluntary food intake was not affected by the change in environmental temperature but the intake of water was reduced to 55% of the thermoneutral value during mild cold exposure and to 40% of the thermoneutral value during the moderate cold stress. 5. It is concluded that a reduction in blood flow to the mammary gland could be one of several possible mechanisms underlying the depression of milk secretion during cold exposure.


Changes in excitability of the cell membrane during 'differentiation without cleavage' in the egg of the annelid, Chaetopterus pergamentaceus. 1. The egg of the polychaete, Chaetopterus pergamentaceus, differentiates parthenogenetically without cleavage after 1 hr in a high K+ solution. The changes in the electrical properties of the membrane during differentiation have been investigated. 2. The treatment with the K solution for 40-60 min made unfertilized eggs become amoeboid cells in 4-5 hr and finally ciliated unicellular embryos in 14-16 hr. 3. In the untreated egg the action potential is Ca dependent and no Na component is found. The steady-state current-voltage relation has a marked inward rectification and shows a less marked outward rectification. 4. There are no significant changes in these properties of the cell membrane immediately after 40-60 min K treatment. 5. In the amoeboid cell stage (4-5 hr) the outward rectification increases significantly. 6. In the ciliated unicellular embryo the action potential is Ca and Na dependent. 7. The result suggests that Ca channels are present in the egg initially, K channels appear next and Na channels appear later.


Effects of postnatal corticosterone treatment on reproductive development in the rat. Reproductive development was studied in rats in which corticosterone (25 mg/100 g body wt) mixed with an equal amount of cholesterol was implanted i.p. at Days 3, 6, 12 or 18 after birth. Vaginal perforation in rats treated on Days 12 or 18 was advanced by 10 days but reproductive function in the treated animals was generally disrupted. A variable incidence of persistent vaginal oestrus was observed in adult females given implants on Days 3 and 6, and of prolonged vaginal dioestrus in those treated on Days 12 or 18. All treated females had lordosis quotients lower than those of untreated controls, the effect being most pronounced at the earlier ages of treatment. The results are believed to indicate two periods in reproductive development which are differentially affected by corticosterone.


Effects of prostaglandins on ovarian blood flow in the bitch. Oestrus and anoestrous mongrel dogs were anaesthetized with chloralose-urethane. In one group, the ovaries were isolated in situ and the effects of a 15-min infusion of PGF-2alpha or PGE-2 on perfusion pressure were measured. In the other group, heated thermocouples were introduced into the stroma of each ovary to measure the changes of local blood flow in response to PGF-2 alpha and PGE-2 infused into the ovarian bursa for 15 min. Intra-arterial infusion of 25.50, 100 or 200 ng PGF-2alpha/kg/min did not affect perfusion pressure; PGE-2 doses of 3.1, 6.2, 12.5 or 25 ng/kg/min caused reductions in proportion to the dose. All doses of PGE-2alpha (50.0, 100 or 200 ng/kg/min) or PGE-2 (25, 50 or 100 ng/kg/min) increased blood flow in the ovarian stroma in proportion to the dose when administered by infusion into the ovarian bursa. There were no differences in the results from oestrous and anoestrous dogs. It is concluded that PGF-2alpha changes intraovarian blood distribution without interfering with the total blood flow while PGE-2 increases both the total and local ovarian blood flow.


Oestradiol-17beta, progesterone, FSH and LH in prepubertal calves induced to superovulate. Fluorogestone acetate (vaginal sponge for 4 days) and PMSG (i.m. injection at the time of sponge insertion) treatment was administered to seven 3-month-old calves to induce superovulation. Samples of peripheral plasma were taken every 4 h during treatment (4 days) and then every 2 h for 7 days. FSH, LH, oestradiol and progesterone were measured by radioimmunoassays. In all calves oestradiol concentrations increased 24 h after PMSG injection and reached the highest levels (41-502 pg/ml) during the preovulatory surge of both gonadotropins. The surge of LH and FSH occurred from 12 to 22 h after cessation of treatment. The maximum levels of LH and FSH were 11-72 ng/ml and 23-40 ng/ml respectively and occurred within 4 h of each other. Between 40 and 68 h after the LH peak the concentrations of progesterone began to increase from basal values, reaching 24.0-101.7 ng/ml when the animals were killed. A quantitative relationship was found between plasma oestradiol concentration and the numbers of ovulating follicles. Progesterone levels seemed to be related to the numbers of corpora lutea and also to the numbers of unovulated follicles. Gonadotrophin output was not quantitatively related to ovarian activity or to steroid secretion.


Fertility, ovulation and maturation of eggs in mares injected with HCG. Pony mares were observed from January to August for incidence of oestrus, duration of oestrus, length of the oestrous cycle and for ovulation and fertility after injection of HCG. From January to 15 May most mares showed oestrus but the duration of oestrus was quite variable and few mares ovulated in response to HCG. From 15 May to 17 August oestrous cycles were more regular and ovulation was induced within 40-50 h by an intramuscular injection of 1500-5000 i.u. HCG. Pregnancy was established by one mating at a fixed time after HCG in 20 of 69 mares. Degenerate eggs were recovered from the oviducts of anoestrous recently ovulated, mated, unmated and pregnant mares. The first polar body was formed before ovulation in 2 eggs and had not formed in 2 recently ovulated eggs flushed from the oviduct. The second polar body formed after sperm penetration 10-12 h after ovulation. After formation of pronuclei, the first cleavage division occurred at 20 h and the second at 32 h after ovulation. Oestrus was inhibited by progesterone administered by vaginal devices but occurred within 1-3 days in 12 of the 20 mares after withdrawal of the devices.


Genetic and non-genetic effects on plasma LH concentrations in lambs at 4 and 8 weeks of age. Plasma LH was measured at 4 and 8 weeks of age in purebred lambs born to Finnish Landrace, Galway, Fingalway and High Fertility ewes. The average litter sizes for these breeds vary from 1.4 to 2.5. Lambs born after the transfer of fertilized eggs (all Fingalway) to Finnish Landrace, Galway and Fingalway ewes were also studied. There was evidence for a significant positive effect of selection for litter size within the Galway breed on plasma LH at 8 weeks of age. Finnish Landrace lambs had significantly higher levels of plasma LH at 8 weeks of age than did Galway Control lambs. These two groups did not differ with respect to plasma LH at 4 weeks of age. Non-additive genetic effects were significant at 4 weeks of age. The ranking of genetic groups on plasma LH was not consistent with their ranking for litter size. There was no evidence for a maternal effect associated with breed of dam on plasma LH and the effects of litter size were generally insignificant. Female lambs had significantly higher levels of LH at 4 weeks of age but there was no sex difference at 8 weeks because LH levels in males had increased. The correlation between the level of plasma LH at 4 and 8 weeks of age was -0.04. It is concluded that plasma LH is unlikely to prove a practical aid to selection for increased fecundity in sheep.


Seasonal variations in post-partum plasma progesterone levels and conception in primiparous and multiparous dairy cows. Progesterone concentrations in peripheral plasma of 18 primiparous and 34 multiparous dairy cows were determined by radioimmunoassay every 4 days, from 10 days post partum until the next conception. The interval from parturition to the first progesterone peak (greater than 4 ng/ml plasma) was significantly longer (P less than 4 ng/ml plasma) was significantly longer (P less than 0.01) in primiparous than in multiparous cows. The progesterone concentrations on Days 4-15 of the oestrous cycle were significantly lower (P less than 0.025) during the summer than in cycles occurring in winter. The conception rate during the summer was lower than in winter. In cows inseminated in summer, conception was related to the shape of the progesterone curve in the cycle preceding insemination.


Effect of prostaglandin F-2alpha and uterine extracts on progesterone secretion in vitro by superfused pig corpora lutea. Corpora lutea obtained from sows in the mid-luteal phase of the oestrous cycle were superfused with various doses of PGF-2alpha, uterine flushings or superfusates from endometrial strips. High doses of PGF-2alpha (1-5 microgram/ml) gave transient stimulation of progesterone secretion; lower doses (0.1-1.0 microgram/ml) inhibited secretion of the steroid, but to a maximum of 55% only and the secretion returned to control levels when superfusion with PGF-2alpha stopped. Uterine flushings from sows in the late luteal phase of the cycle were more effective than mid-cycle flushings in inhibiting progesterone secretion by the luteal tissue: maximal inhibition was 65% and progesterone secretion slowly returned to control levels when superfusion with the flushings was stopped. Superfusate from strips of uterine endometrium from sows in the late luteal phase of the cycle proved most effective in inhibiting progesterone secretion (greater than 90%) and gave a long-lasting inhibition which did not appear to be due to stimulation of PGF-2alpha secretion by the endometrium or corpus luteum.


Seasonal variation in oestrus, the secretion of oestrogen and progesterone and LH levels before ovulation in the ewe. Ewes with cervical ovarian autotransplants were studied after PGF-2alpha-induced luteal regression in November and February (mid- and late-breeding season) and June (anoestrum). Progesteron and oestradiol-17beta secretion rates and LH concentrations were determined in serial ovarian venous blood samples and the ewes were frequently tested for oestrus. All 4 ewes in November and 3 of the 4 ewes in February exhibited oestrus and endocrine changes indicative of ovulation. The remaining ewe in February and the three ewes in June failed to show elevated oestradiol-17beta secretion rates after luteal regression, indicating the absence of follicles in the final stages of maturation. The preovulatory rise in the oestradiol-17beta secretion rate, the LH surge and the display of oestrus all occurred earlier, with respect to the PGF-2alpha infusion, in November than in February, suggesting a greater stimulation of folliculogenesis, and therefore a greater availability of maturing follicles, in November than in February.


A novel synthesis and biological activity of several 5-halo-5'-amino analogues of deoxyribopyrimidine nucleosides. A novel synthetic procedure has been developed for the large-scale synthesis of 5-chloro-, 5-bromo-, and 5-iodo-5'-amino-2',5'-dideoxyuridine (4c-e) as well as of two new analogues, 5-iodo-5'-amino-2',5'-dideoxycytidine and 5-fluoro-5'-amino-2',5'-dideoxyuridine (4a and 4b), in good yield. The starting materials, 5-halo-2'-deoxyuridine and 5-halo-2'-deoxycytidine, are readily available and the method is straightforward. This report describes the synthesis and the biologial activities of these compounds.


Synthesis and biological activity of several amino analogues of thymidine. 3',5'-Diamino-3',5'-dideoxythymidine (7) was synthesized via a nine-step synthesis from thymidine in good overall yield. 3'-Amino-3'-deoxythymidine (8) and 5'-amino-5'-deoxythymidine (12) were prepared with a minor modification of the procedure reported by Horwitz and co-workers. Although the 5'-amino analogue 12 had potent antiviral activity relative to the 3'-amino analogue 8, the latter is a potent inhibitor of the replication of both murine sarcoma 180 cells (ED50 = 5 micrometer) and of murine L1210 cells (ED50 = 1 micrometer) in vitro. Most unexpectedly, however, was the finding of complete lack of either antiviral or antineoplastic activity by the 3',5'-diamino analogue 7 which appears to have acquired the undesirable qualities of both the 3'-amino and 5'-amino analogues of thymidine.


Antineoplastic agents. 1. Synthesis and antineoplastic activities of chloroethyl- and methylnitrosourea analogues of thymidine. A new class of chloroethyl- and methylnitrosourea analogues of thymidine, 5a,b, 6, 10, and 11, has been synthesized from the corresponding amino nucleosides, 2 and 7. The 3'-chloroethyl and 3'-methyl derivatives, 10 and 11, inhibited L1210 cell growth in culture (ED50 = 1.5 and 1.0 micrometer, respectively) more effectively than 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) (ED50 = 4 micrometer) and the 5'-nitrosourea analogues. Neither the alkylating nor the carbamoylating activities of these compounds correlated with their biological activity.


Structural requirements for antileukemic activity among the naturally occurring and semisynthetic maytansinoids. In an effort to determine the structural requirements for the significant antileukemic, cytotoxic, antitubulin, and antimitotic activity exhibited by the novel ansa macrolide, maytansine (1), four new C-3 ester and six new C-9 ether homologues were synthesized. The biological activities of these compounds were assayed and compared to the activities of previously reported, naturally occurring maytansinoids. From the data, it is apparent that presence of the C-3 ester is necessary for significant activity, and variations in the ester group are not accompanied by marked changes in activity. However, elimination of the ester group, as in maytansinol (7), maysine (8), normaysine (9), and maysenine (10), results in a significant decrease in biological activity. Blockage of the C-9 carbinolamide via etherification markedly reduces antileukemic and cytotoxic activity and slightly reduces antitubulin activity but has relatively little effect on antimitotic activity against sea urchin eggs. Thus, a free carbinolamide at C-9 is advantageous for optimal activity.


Predisposition to spina bifida: Search for a relation to maternal gastric acid secretion. The peak acid output of the stomach in a group of 71 mothers of spina bifida children and in 71 matched controls was estimated indirectly by the serum level of group I pepsinogens. The mean levels did not differ significantly, suggesting that the conjectured teratogen is not specially acid-labile. The variance was significantly higher in index subjects than in controls, but the interpretation of this finding is not clear.


Testicular feminisation syndrome: unusual gonadal histology in an elderly patient. The gonads of an elderly patient with a typical testicular feminisation syndrome are described. The unusual histological features consisted of total absence of testicular tubular structures or remnants thereof and distinct proliferation of smooth muscle bundles. In addition, there was fibrous proliferation, areas of ovarian stroma, and rare Reinke crystalloids within Leydig cells. The complete tubular absence may have been the result of fibrous replacement related to patient's advanced age, while the muscular proliferation may have been of hamartomatous nature. Thus, it seems that in elderly patients with testicular feminisation syndrome, the histological appearance of the gonads may vary considerably from that in younger individuals, and in such cases the correct diagnosis should be based mainly on clinical and cytogenetic findings.


Spermatogenesis and spermiogenesis in Ascaris lumbricoides Var. suum. Reorganization of the prophase I nucleus marks the beginning of the first meiotic division. A pair of centrioles is present at each pole at metaphase I and mitochondria are not observed in the spindle area. A chromosomal pellicle, which resembles a kinetochore plate but has no apparent association with microtubules, surrounds each autosome at metaphase I and II. The sex body lags behind the autosomes at anaphase I and segregates differentially to one daughter cell. Mitochondria and a pair of centrioles are present in the spindle during the second meiotic division. Localized condensation of chromatin and fusion of the condensed chromatin of the secondary spermatocyte telophase nucleus results in a compact spermatid nucleus. Loss of spermatid cytoplasm is effected by the ejection of a cytophore vesicle.


Children at birth: effects and implications. The sexual attitudes and beliefs of 20 children who have been present at the labor and delivery of sibs and have observed the birth process are compared with 20 children who have not been present at delivery. The ages of these children ranged from 2 years to 14 years. Several of these children were observed at the actual delivery where naturalistic observations of delivery were carried out. Naturalistic observations of these processes were also carried out as well as observations of play of children who had witnessed the delivery with children who had not. The attitudes of the parents of children present at delivery were characterized and consisted of a feeling that birth was a normal family event of a positive nature that should involve other children of the family. Their children tended to have very accurate notions regarding the proverbial "Where do babies come from?" question and tended to view the birth in a positive, happy manner. Children not present found the idea of birth puzzling or inconceivable and did not seem to understand the concept or be able to relate it to their family. They tended to develop theories about birth that were inaccurate and sometimes fantastic, much as those described earlier by Freud. The results seem to suggest that children's presence at delivery permits an open attitude toward birth as a normal process in families in which this ideology is held and for girls aids in the development of their self-image as women. Preliminary data suggests less sibling rivalry among the children present at the delivery. These results are discussed in regard to current American birth practices.


Fortification variability in rat diets fortified with arsenic, cadmium, and lead. Eleven rat diets fortified with 200 ppm Pb as lead acetate, and/or 50 ppm Cd as cadmium chloride, and/or 50 ppm As as sodium arsenate or p-arsanilic acid for use in a feeding study to determine the interactive effects of the three elements in rats were analyzed to confirm the fortification levels. Gross inhomogeneity was found. After remixing, variability within and among diets was improved but was still unacceptable. Fortification levels of Pb and of As as sodium arsenate could be accurately determined only by taking a very large sample and homogenizing the diet subsamples with a Polytron homogenizer. A review of the recent literature indicates that no standard procedures exist for fortification, mixing, or analysis of animal feeding study diets. Such procedures should be developed and analysis of the diets before and at intervals during a feeding study should be adopted as standard procedure.


The prevalence of human intestinal protozoa in Ibadan, Nigeria. This study shows that the two intestinal protozoa, Giardia lamblia, Balantidium coli in addition to Entamoeba histolytica are prevalent in the Ibadan population and are responsible for many of the non-bacterial diarrhoea seen in patients. A high seasonal prevalence of the intestinal protozoa in the dry months of the year was associated with the use of contaminated water. Both the high infection rates recorded in adults and children under five years old, and the high frequency of association involving Trichomonas hominis, Entamoeba histolytica and Giardia lamblia were discussed.


Binding characteristics of Rauscher leukemia virus envelope glycoprotein gp71 to murine lymphoid cells. The major envelope glycoprotein (gp71) purified from Rauscher leukemia virus (R-MuLV) binds efficiently to murine lymphoid cells but not to either murine nonlymphoid cells or lymphoid cells from other species. Binding of 125I-labeled R-MuLV gp71 was competitively inhibited by unlabeled glycoprotein, as well as by whole R-MuLV, but not by murine xenotropic viruses, R-MuLV p30, and several unrelated proteins. Polyacrylamide gel electrophoresis profiles of iodinated gp71 after binding to lymphoid cells were similar to prebound profiles. Antibody to R-MuLV gp71 prevented binding, whereas normal serum had no effect. Adsorption of the glycoprotein to murine lymphoid cells occurs rapidly and is time and temperature dependent. The procedure described is sensitive for detecting the binding activity of approximately 10(4) cells. Binding was proportional up to 2.5 X 10(5) cells per ml and plateaued above 10(7) cells per ml. In the presence of excess R-MuLV gp71, BALB/c thymocytes bound approximately 2.4 X 10(4) molecules per cell.


Peptide map comparison of infectious pancreatic necrosis virus-specific polypeptides. An investigation of virus-specific protein synthesis in infectious pancreatic necrosis virus (IPNV)-infected rainbow trout gonad cells was undertaken to find a relationship between the coding capacity of the virus genome (two segments of double-stranded RNA of 2.5 x 10(6) and 2.3 x 10(6) molecular weight) and the sizes and relative amounts of virus-specific proteins. Using polyacrylamide slabgel electrophoresis and autoradiography, eight distinct virus-specific polypeptides were detected in infected, [(35)S]methionine-labeled cells. These proteins may be grouped into three size classes on the basis of molecular weight: (i) large, alpha (90,000); (ii) medium, beta(1) (59,000), beta(2) (58,000), and beta(3) (57,000); and (iii) small, gamma(1) (29,000), gamma(1A) (28,000), gamma(2) (27,000), and gamma(3) (25,000). The combined molecular weight of these polypetides (373,000) is beyond the coding capacity of the virus genome. Purified IPNV contained polypeptides alpha, beta(3), gamma(1), and gamma(1A). Pulse-chase experiments and tryptic peptide map comparisons revealed that only four of the eight intracellular proteins were primary gene products, namely, alpha, beta(1), gamma(1), and beta(2), with a combined molecular weight of 205,000. Of these primary gene products only the alpha polypeptide was found to be stable, whereas the other three underwent intracellular proteolytic cleavage during virus morphogenesis. Polypeptide beta(1) was cleaved to generate beta(2) and beta(3); gamma(1) was trimmed to produce gamma(1A), and the only nonstructural primary gene product, gamma(2), was found to be a precursor of gamma(3). These results suggest that IPNV possesses a unique mechanism to synthesize three size classes of proteins using mRNA transcripts from two high-molecular-weight double-stranded RNA genome segments.


Myocardial infarction due to metastasis of choriocarcinoma in a 29-year-old woman. A rare and instructive case of myocardial infarction in a 29-year-old woman was presented. In this case it was clinically suspected that myocardial infarction was due to metastatic choriocarcinoma of the heart. This was confirmed by autopsy. It was emphasized that metastatic choriocarcinoma of the heart could be a cause of myocardial infarction in young women.


Hepatic aminopyrine N-demethylase system: effect of cyanide on microsomal N-demethylase activity. Cyanide, an inhibitor of many hemoproteins, was shown to affect a number of microsomal drug-metabolizing activities catalyzed by cytochrome P-450. The N-demethylation reaction of aminopyrine was inhibited noncompetitively by this inhibitor in microsomal preparations from rats. The binding reaction of aminopyrine with microsomal cytochrome P-450 was also modified by cyanide, and an abnormal aminopyrine-induced difference spectrum of microsomes by cyanide, and an abnormal aminopyrine-induced difference spectrum of microsomes appeared when cyanide was added to the reaction mixture. Partial dissociation of cytochrome P-450. Cyanide complex by aminopyrine was observed by spectrophotometrical and epr spectroscopic methods. These results suggest that aminopyrine and cyanide reciprocally affect binding with cytochrome P-450 and modification by cyanide of aminopyrine binding reaction with the hemoprotein produces an inhibition of N-demethylase activity.


Unusual aggressive behavior in the male golden hamster. An unusually aggressive male golden hamster (Mesocricetus auratus) removed flank glands from other adult male hamsters by biting. Females and immature males were not attacked. No underlying abnormality to account for this aberrant behavior was evident.


[Effect of seduxen on the course of experimental hypokinesia]. It has been found that seduxen at a dose of 5-10 mg/kg/day administered in different ways to hypokinetic animals does not prevent hypoxic developments in the myocardium and the femoral muscle or does not normalize hypokinesia-induced biochemical changes in the liver. These findings make the advisability of seduxen application during hypokinesia doubtful.


Hexose transport regulation in cultured hamster cells. Hamster (nil) cells maintained overnight in culture medium containing cyclohemiximide and either glucose or fructose exhibit strikingly different rates of hexose transport and metabolism (i.e. uptake). Pretreatment of cultures with sulfhydryl reagents makes it possible to determine initial transport rates for a physiological sugar such as galactose which is a catabolite in hamster cells. Using galactose transport as a model, hexose uptake enhancements can now be shown to be due almost entirely to increases in the rate of the transport step. The transport regulation can best be accounted for by a model comprised of 2 antagonizing mechanism. This model involves turnover of transport carriers as well as inhibitory units ("regulators"). The experimental as well as the theoretical model may also apply to the well-known uptake enhancements observed in oncogenically transformed cells.


Distribution of a major surface-associated glycoprotein, fibronectin, in cultures of adherent cells. Fibronectin was present in media and cell layers of cultures of adherent cells from human skin, kidney, lung, chest wall, liver, and heart. Cell-surface fibronectin, visualized by immunofluorescence, was in dense fibrillar (cultures from lung), discrete fibrillar (e.g., cultures from skin), or punctate (some cultures from kidney) structures. The subunit sizes of cell-surface fibronectin and fibronectin soluble in medium appeared identical in sodium dodecyl sulfate-polyacrylamide gels. To explain the polymorphism of cell-surface fibronectin, there must be chemical differences among the fibronectins synthesized by different cell strains or factors in the cell layer which influence fibronectin binding and aggregation.


Characterization of the Fc receptors of the murine leukemia L1210. A glycoprotein extract prepared from the plasma membranes of L1210 cells was passed over columns of Sepharose 4B to which either heat-aggregated human IgG or F(ab')2 fragments has been coupled. The intact IgG column bound 35.7 percent of the applied counts, whereas the F(ab')2 columns bound 2.8 percent. The bound glycoproteins were eluted with citrate buffer (pH 3.2) and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Three peaks with apparent molecular weights of 65,000 45,000, and 28,000 daltons were identified and purified by electroelution from polyacrylamide gels. The isolated proteins were able to bind to the same sub-classes of mouse IgG myeloma proteins as the intact L1210 cells, indicating that these molecules are related to L1210 surface Fc receptors. Amino acid analyses of the 3 proteins were markedly similar suggesting that the observed molecular heterogeneity might be due to carbohydrate differences. Neuraminidase digestion of the isolated proteins resulted in mobility shifts on polyacrylamide gel electrophoresis which were consistent with the interpretation that either the isolated proteins have considerably different sialic acid contents, or that removal of the sialic acid results in disaggregation of an Fc receptor molecule.


Characterization of the alcohol research literature. Bibliometric analyses of the inernational, rapidly increasing and widely dispersed alcohol research literature show three major components: biomedical, biosocial, and psychosocial research; the biomedical component predominates.


Effects of shock and darkness on alcohol consumption by rats. Alcohol consumption by three groups of rats, one subjected to electric shocks, one placed in darkness and a control group, increased as a function of time.


The cyclic changes of the lymph vessels in the uterus of the mouse which has been pregnant. When adapted to the natural possibility of reabsorption of the uterus, using the dye Patent Blue Violet and Japanese ink, the orthograde demonstration of the cyclical behaviour of the lymphatics during the oestrus cycle was successful even with parous mice. As investigations have already shown in nulliparous mice (5), the characteristic variations in all five stages of the sexual cycle are also found here: As a result of the pregnancies the lymphatics of the endometrium are more strongly developed, in contrast to the nulliparous mouse. However, in the stage of pre-oestrus and oestrus the marked or very marked widening, of varying calibre does not show in such a striking way. In the subsequent two stages of the secual cycle prominent elevations are found in the lumen of the uterus, and these have lymphatic capillaries running in them. The further stage, a more or less reticulate arrangement of the lymphatic capillaries, with fine branching ramifications, is equally typical. As a result of the pregnancies, the larger lymphatic vessels are often curled up like a cork-screw, and also show their typical beaded appearance.


Effects of lectins on the hemolysis of rabbit erythrocytes by straphylococcal alpha toxin. When concanavalin A (1 microgram/ml) or wheat germ agglutinin (2 microgram/ml) was preincubated with a suspension of 2% rabbit erythrocytes for 5 min at 20 C, the binding [125I]-labeled staphylococcal alpha toxin to these erythrocytes was greatly inhibited and the hemolytic action of alpha toxin was decreased. The inhibitory effect of concanavalin A on hemolysis by alpha toxin was completely reversed in the presence of 0.1 M alpha-methyl-D-glucoside or alpha-methyl-D-mannoside. Phytohemagglutinin-P from Phaseolus vulgaris and soybean agglutinin inhibited hemolysis by the toxin at concentrations exceeding 20 microgram/ml. The effect of concanavalin A on alpha-toxin hemolysis was studied further to ascertain the nature of the inhibition. Double reciprocal plots were made of hemolysis against alpha toxin concentrations, and the data suggested that inhibition of the initial rate of the hemolysis by concanavalin A is competitive in nature. This was probably due to an interaction with the alpha toxin binding sites on the cell membrane surface.


Simulated patients as a learning resource in the study of reproductive medicine. Simulated patients have been used to foster active learning and problem solving skills in an undergraduate programme of reproductive medicine. These simulators were trained to present a series of clinical problems and were used in a variety of approaches. They served to highlight communication skills and to force attention on the recognition, assessment and management of  patient problems.


An outbreak of phaeohyphomycosis in rainbow trout caused by Scolecobasidium humicola. A previously undescribed host for the opportunistic dematiaceous hyphomycete, Scolecobasidium humicola, is reported. Several epizootics among rainbow trout, Salmo gairdneri, occurred in a Tennessee fish hatchery from 1969 to 1973. Symptoms included surface lesions, blisters and abscesses. The kidneys and other internal organs were invaded by the mycelium of S. humicola. Tissue morphology of the fungus was typical of that associated with phaeohyphomycosis Experimental infections were reproduced in fingerling rainbow trout after intraperitoneal inoculation of S. humicola. Following a change in the hatchery's water supply, no new epizootics have occurred.


Regulation of protein synthesis, intracellular electrolytes and cataract formation in vitro. Control of protein synthesis is associated with changes in the ratio of intracellular Na+ to K+ in the cultured embryonic chick lens. Correlations of intracellular Na+/K+ ratios with crystallin synthesis and cataract formation in vitro suggest that the Na+/K+ ratio may have an important role in the regulation of protein synthesis during cataractogenesis.


[Transmembrane ionic currents in neuroblastoma cells]. Isolated neuroblastoma cells (clone A-1 from clone N-18) were investigated by means of the intracellular dialysis technique. Changes in the ionic composition of intra- and extracellular media show that the fast inward current is carried by Na ions and delayed outward current is carried by K ions. Na inward current was blocked by TTX. Calcium inward current was not observed. Substitution of Na ions for K ions showed that PNa: PK = 7:1. A relatively low potassium conductance was found in the neuroblastoma cells.


Results of dynamic dose-fractionation combined with breathing oxygen in comparison to conventional treatment in head and neck and uterine cervix cancer. A dynamic dose-fractionation regime combined with breathing oxygen at ambient pressure was introduced in the treatment of head- and neck cancer, and uterine cervix cancer. The efficacy of the method was analyzed in relation to the results obtained by treating comparable cases with conventional fractionation regime. Final evaluation of the method will have to await the accumulation of further experience during a prolonged observation period. Encouraging results obtained so far suggest that the new treatment can be accepted as a possible improved routine in the therapy of some cancer sites.


[Clinical evaluation on glucocorticoid administration in the neurosurgical patients (the third report)--serial plasma cortisol assay following betamethasone 8 mg intravenous injection (author's transl)]. The effect of betamethasone on plasma cortisol concentrations was studied in 6 healty adult men. Each of them was administered 8 mg of betamethasone in a single intravenous injection. Blood samples were obtained serially for 2 days before the injection of betamethasone and for a week after the injection. Plasma cortisol was measured by competitive protein binding analysis. Before the injection of betamethasone plasma cortisol levels showed the normal diurnal rhythm in all the cases. After the injection plasma cortisol levels decreased rapidly and a mean half life time was about 1.7 hours. The low levels of plasma cortisol under 2 microgram/ml were observed for 48 hours after the injection, and then the diurnal rythm of plasma cortisol reappeared, but the levels of plasma cortisol at this time were still lower than the control levels. Then plasma cortisol levles increased gradually and they returned to the control levels 5 days after the injection. Thus, it was clarified that the suppressive effect of 8 mg or betamethasone on plasma cortisol concentrations lasted at least for 48 hours. From this point of view the time interval or repetitive glucocorticoid administration for the treatment of brain edema will further be studied.


Stimulation of the Hypothalamus and FSH and LH secretion in the ferret. The acute effects of hypothalamic electrical stimulation on the secretion of FSH and LH have been examined in anestrous and estrous ferrets. Basal levels of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were higher in the anestrous than in the estrous animals; the anestrous animals showed a more marked increase in gonadotropin levels following hypothalamic stimulation than did the estrous animals whether the stimulation current was constant or increased in the course of the experiment. It is concluded that the physiological difference in the control of gonadotropin secretion in the estrous (vs the anestrous) ferret is not reflected in an increased sensitivity of the hypothalamo-hypophysial axis to stimulation.


Return of diurnal plasma corticosterone rhythm long after frontal isolation of the medial basal hypothalamus in the rat. The diurnal rhythm of plasma corticosterone (B) in adult male rats was investigated 6, 12, 18, 24, 30 and 36 weeks after frontal isolation of the medial basal hypothalamus (MBH) and was compared to that of sham-operated controls. Taking blood samples by heart puncture, it was possible to follow the same groups of animals during the entire experimental period. In accordance with earlier reports, frontal deafferentation of the MBH abolished the diurnal rhythm of plasma B 6 weeks after surgery. No change was observed in this respect until the 30th postoperative week. At that time, however, the rhythm tended to return and was present by the 36th week in the operated animals.


[Late neurological sequelae of tick-borne encephalitis in the population of the Olsztyn province]. The authors studied the incidence of late and permanent neurological sequelae of tick-borne encephalitis in the population of the Province of Olsztyn. The investigations were carried out in a group of 103 patients who were affected in the years 1970--1974. The diagnosis was confirmed in all cases with epidemiological, clinical and laboratory investigations and serological evidence of tick-borne encephalitis. It was shown in these studies that tick-borne encephalitis caused development of permanent lesions in the central and peripheral nervous system in 25.3% of cases. It was observed, moreover, that a considerable part of neurological signs found in the acute stage of the disease disappear during hospital treatment but later on after discharge from the hospital the symptoms of nervous system damage reappear and new signs develop which had not been observed in the acute stage. In the light of these studies the authors conclude that a considerable proportion of convalescents have limited physical and intellectual efficiency making impossible their return to their previous work and causing not infrequently permanent loss of working ability which must be kept in mind by physicians from different specialities.


The effect of denervation on compensatory ovarian hypertrophy. Hemiovariectomized rats were randomly assigned to 1 of 5 groups: controls, 6-hydroxydopamine (6-HD)-treated, abdominal vagotomy, 6-HD-treated plus abdominal vagotomy and pelvic parasympathectomy. 15 days later all animals were sacrificed and the amount of compensatory ovarian hypertrophy (COH) was calculated. Vagotomy and vagotomy plus 6-HD treatment interrupted estrous cycles and significantly decreased COH. Vagotomized rats with both ovaries intact had disrupted estrous cycles but ovarian weights were not affected. In a subsequent study, rats in estrus were sham-operated, unilaterally ovariectomized (ULO), vagotomized, or vagotomized + ULO, and serum levels of LH and FSH were determined at 5 and 24 h. ULO caused a significant (p less than 0.05) increase in LH and FSH at 5 h. Vagotomy significantly (p less than 0.05) depressed LH and FSH levels in hemiovariectomized animals at 5 h. By 24 h LH was significantly higher in ULO than in either sham-operated (p less than 0.05) or vagotomy (p less than 0.01) groups. Also, vagotomy significantly (p less than 0.01) depressed FSH levels at 24 h. These results suggest a functional role for the vagus nerve in normal cyclic activity, COH, and gonadotrophin (Gn) secretion.


Ambulatory neurology in residency training programs: a perspective. Inpatient care and teaching have been the central theme in academic medicine, including neurology, for the last several decades. However, pressures from within and outside of medicine are building to create a renewed commitment to ambulatory care and education. Most neurology training programs have not yet made that commitment. They should do so for the benefit of both the patients and residents served by the programs. Effective outpatient clinic training programs should include full-time staff devoted to ambulatory care, teaching, and research. The practicing clinical neurology staff could be used more effectively. Regular chart review, case management conferences, and other teaching strategies, and the reorganization of the clinic to include private patients, as well as group practice and interdisciplinary team concepts, could improve both the patient care and resident training in the neurology clinic.


Narrowing and occlusion of arteries by intracranial tumors. Review of the literature and report of 25 cases. The intracranial arterial obstructions (narrowing or occlusion) found with intracranial tumors are rarely reported. Our 25 cases and those of the literature enable us to draw a statistical view of these facts. Meningiomas and gliomas are often concerned, and they principally involve the main arteries at the base of the brain. The practical interest in such pathology concerns: the risk of a fall in blood pressure during general anesthesia which may result in occlusion of a narrowed artery; the need to estimate the pathological state of one or several arteries passing by a tumor which must be removed; and the possibility of providing an extra-intracranial arterial bypass if necessary.


Diaphragmatic paralysis in motor neuron disease. Report of two cases and a review of the literature. Two patients presenting with respiratory insufficiency had diaphragmatic paralysis secondary to adult-onset motor neuron disease (progressive spinal muscular atrophy). A review of the literature discloses seven similar cases, most of them reported in non-neurological journals.


Kindling of the mesolimbic dopamine system: animal model of psychosis. In order to examine the behavioral and physiologic consequences of chronic activation of the mesolimbic dopamine system, the nucleus of origin in the ventral tegmental area was stimulated electrically for 2 seconds daily through chronically implanted intracranial electrodes in cats at the same point where instillation of the GABA blocking agent bicuculline induced a characteristic fear, staring, searching, and withdrawal response. None of the animals developed sustained after-discharge or seizures following daily stimulation for 2 months. Progressive fearfulness, hiding, loss of social behavior, and EEG spike or slow activity in the ipsilateral nucleus accumbens developed in three of six intact animals. Two cats with prior 6-hydroxydopamine lesions of catecholamine pathways did not develop behavioral change in response to local bicuculline or daily electrical stimulation of the ventral tegmental area but demonstrated pronounced after-discharge or EEG spike propagation during the kindling procedure.


Vitamin B6-dependent seizures: pathology and chemical findings in brain. A 13 1/2-year-old child died with vitamin B6-dependent seizures in progress. Microscopic findings in brain included an abnormally sparse quantity of central myelinated fibers in the cerebral hemispheres. Glutamic acid concentrations were elevated and GABA concentrations reduced in the frontal and occipital cortices but not in the spinal cord. All other amino acid concentrations were normal, except for increased cystathionine in the occipital cortex. Pyridoxal-5-phosphate (PLP) was reduced in the frontal cortex. Glutamic acid decarboxylase activity comparable to that of controls was detected when the PLP concentration was greater than 0.05 mM. These findings suggest that pyridoxine-dependent seizures in man are associated with reduced GABA concentrations in the brain and with diminished central white matter structures.


Potentiation and antagonism of serotonin effects on intracranial and extracranial vessels. Possible implications in migraine. The vasoconstrictor action of serotonin and its interaction with drugs (methysergide, pizotifen, cyproheptadine, ergotamine, dihydroergotamine) that have serotonin-antagonistic properties and that are used in the treatment of migraine were studied on isolated feline and human intracranial and extracranial arteries in vitro. The intracranial vessels were more sensitive to serotonin than the extracranial ones. The serotonin antagonists caused the expected impairment of the serotonin-induced contraction. In minute doses, the various compounds potentiated the contraction produced by serotonin. There is reason to believe that the mechanism responsible for this interaction is selective for tryptaminergic agents. This dual action of the serotonin antagonists may contribute to a beneficial effect in migraine by an interference during both the vasoconstrictory and vasodilatory phase.


Pediatric metrizamide CT cisternography: cerebrospinal fluid circulation and hydrocephalus. The technique of intrathecal enhancement using metrizamide (Amipaque) combined with CT imaging enables both the morphologic and dynamic (serial) evaluation of the intracranial cerebrospinal fluid pathways. Three distinct metrizamide CT cisternographic patterns (delayed, intermediate, and normal) emerge in pediatric patients with hydrocephalus. The findings correlate well with radionuclide cisternographic patterns; however, CT also provides excellent morphologic definition.


Status epilepticus in pregnancy: effect of phenytoin malabsorption on seizure control. In the second trimester of pregnancy in a 26-year-old woman, marked exacerbation of epileptic seizures occurred with somatomotor status epilepticus. The oral requirement of phenytoin varied, and up to 1,200 mg per day were needed to maintain a therapeutic plasma concentration during the second trimester. Intestinal malabsorption was shown to be a causal factor; 56 percent of the daily oral dose of phenytoin was found in the stool. Late in pregnancy and postpartum, therapeutic plasma concentrations of phenytoin were maintained with decreased daily oral doses. Intestinal absorption improved postpartum.


Prevalence of epilepsy in Bogota, Colombia. A combined epidemiologic, clinical, and electroencephalographic study of epilepsy was undertaken in Bogota by the Neurological Institute of Colombia, the Colombian Institute of Health, and the Department of Health of the City of Bogota. The prevalence of epilepsy on December 31, 1974, was 19.5 per 1,000, the highest rate for general populations reported in the literature. The prevalence was significantly higher for females than for males, 22.9 per 1,000 compared with 15.5 per 1,000.


Quantitative determination endogenous acid phosphatase activity in vaginal washings. Vaginal washings obtained in 98 patients were analyzed for acid phosphatase activity. A history of recent sexual intercourse was also obtained. Acid phosphatase values of less than 50 IU (obtained by the alpha naphthol phosphate method) correlated with abstinence from sexual intercourse for more than 30 hours, whereas values greater than 50 IU correlated with sexual intercourse in a period up to and including 30 hours (P less than 0.05). The presence or absence of sperm was found to correlate poorly with the time since intercourse.


Diagnostic significance of summating potential in sensorineural deafness. The polarity of the summating potential wave induced by electrocochleographical examination has an important bearing upon the pathophysiology or the prognostic evaluation of sensorineural deafness. Electrocochleographical examination was performed on cases of sudden deafness, Ménière's disease, progressive sensorineural deafness, sensorineural deafness due to barotrauma, etc. When the polarity of the summating potential showed an increased negative wave, physiological blockage in sensorineural structures was revealed. In other words, under such conditions sensorineural structures have a potential for activity although hearing acuity was damaged. On the other hand, when the summating potential has an increased positive polarity wave, gradual decrease of the hearing acuity was revealed.


The facial nerve coursing across the oval window area. A case of unilateral conductive deafness since birth is described. At exploration of the middle ear, the facial nerve was found to have an anomalous course across the oval window region, the stapedial crura ending on the uncovered facial nerve. This case is believed to represent the 9th case to be recorded. This anomaly may be due to failure of union between the branchial part and otic capsule part of the stapes, in combination with a defective development of the laterohyale resulting in a partially absent facial canal.


Automatic audiometry in stapes motility evaluation. An automatic Grason-Stadler-Békésy 1701 audiometer with continuous frequency progression (1 octave/min) and increase (or attenuation)of intensity of 2.5 dB/sec, has been employed to perfect an investigation technique for obtaining, by the mean difference in dB between absolute bone conduction (ABC) and relative bone conduction(RBC), a quantity evaluation of stapes motility and monitor alterations brought on by various disease processes. A preliminary study performed on subjects with normal hearing was used to establish the influence of age on motility. It was thus possible to establish an index (stapes motility index, SMI) to represent the degree of normal stapes motility as a function of the subject's age and thus, through variations in this index, identify alterations to the motility of the tympano-ossicular system caused by ongoing disease processes and by surgery.


Adenomatosis of accessory salivary glands of the lip. Report of two cases. Two cases of an unusual tumour of accessory salivary glands are presented. The histological examination revealed that this tumour was benign and not malignant as has been suggested by others. Clinical follow-up for 8 years showed no signs of recurrence or metastases. We suggested to call it 'adenomatosis'. No extensive surgery is indicated.


The ultrastructure of penetrating stages of Babesia major infecting the ovary of Haemaphysalis punctata. The ultrastructure of penetrating stages of Babesia major merozoites responsible for causing trans-ovarian infection of the tick Haemaphysalis punctata was studied. These merozoites were different from forms of other Babesia species previously described in the tick in that they were highly pleomorphic. Most of the organelles found in these forms were similar to those found in other stages: however, the apical complex was found to be very active. Micronemes were found to be predominant in the cytoplasm of these merozoites and a cytoplasmic projection anterior to the polar ring and bounded by concentric rings was visible. A polar ring with radiating ribs, and microtubules was present. It is thought that the apical complex of these merozoites plays an important role in assisting the parasite to infect the ovary of the tick.


Passive electrical properties of the atrio-ventricular node. The passive electrical properties of the atrioventricular node and of the atrial muscle of the rabbit heart were determined. To polarize the cells, a small suction electrode was used. The space constant of the atrio-ventricular node was found to be small (430 micron) compared to other cardiac fibers. The small value of gamma in nodal cells is due to a high intracellular resistance (ra, 40.9 +/- 9 M omega/cm) which is higher than in atrial muscle (9.6 +/- 2.2 M omega/cm). The input resistance of cells of the N layer was found equals to 880 K omega while in the right atrium was 320 K omega. The time constant of nodal cells in the AN layer was 3.4 ms, in the N layer 9 ms and in the atrium 5 ms. Assuming a specific membrane capacity (Cm) of nodal and atrial fibers of 1 micron F/cm2, Rm was found equals to 9.000 omega cm2 in N layer. 3.400 omega cm2 in AN layer and 3.800 omega cm2 in the right atrium. Acetylcholine (5 microgram/ml) reduced the space constant of the atrio-ventricular node by 38% and the time constant of nodal cells by 33%. The delay of impulse conduction in the A-V node semms then related to a high intracellular resistance along the pathway of conduction.


A multi-sensor support for in vivo monitoring of blood parameters with in situ sensor calibration. An apparatus permitting continuous simultaneous in vivo monitoring of human arterial blood parameters is described. Sensors are held firmly in position in arterial blood flowing through an extracorporeal shunt. The system is reliable, rugged and inexpensive. Installation and manipulation are easy, and the sensors can be calibrated in situ. The system seems well suited for computer-aided monitoring of intensive care patients.


The role of augmented breaths (sighs) in bronchial asthma attacks. The study is based on plethysmographic, neurophysiological and respiratory mass-spectrographic records established during anaphylactic or histamine-induced bronchial asthma attacks in guinea-pigs. The frequency of occurrence of the augmented breaths (sighs) is correlated with the intensity of the lung deflation reflex (tachypnoea) which arises during the attack. In the acute phase of the asthma attack, the sighs increase in number and reinforce the uneven ventilation which underlies stimulation of the pulmonary deflation receptors. The sigh becomes an essential component of the vicious circle of uneven ventilation and vagal lung deflation  reflex induced. This circle is broken in the recovery phase of the asthma attack, during which the sighs decrease in number, as the conditions underlying their occurrence subside; they now lessen the uneven ventilation and by reopening closed lung units promote a return to normal conditions in lung mechanics. It is concluded that augmented breaths, like the asthmatic tachypnoea, are caused by uneven ventilation resulting in pulmonary self-compression and in turn stimulation of the pulmonary deflation (irritant) endings. Some factors which possibly contribute to the elicitation of augmented breaths are discussed.


Fabrication of well defined micropipette tips by hydrofluoric acid etching. Glass tubes were drawn on a two stage pipette puller such that tips with 0.5-2.0 micron inner diameters were obtained as confirmed by scanning electron imaging. The pipettes were attached to an air reservoir with a pressure set at 0.5 bar to prevent fluid from entering the tip. They were lowered into 2% hydrofluoric acid by means of a micromanipulator. When air bubbles began to escape from the tips, they were withdrawn and immersed into 0.5 molar phosphate buffer, pH 7.4, and then rinsed with tap water and ethanol. Etching during 100 s (mean) yielded inner tip diameters of 5 micron (+/- 5% S.D.). The tips were beveled at the same angle at which they had been dipped into the etching fluid. No continuation of the etching process was noticed even after several days.


Equatorial x-ray reflections from contracting muscle after an applied stretch. The equatorial X-ray reflections were recorded from contracting muscle after a slow stretch. The intensity ratio of the 1,0 to the 1,1 reflections (I10/I11) after the stretch was not significantly different from that during an isometric tetanus at the same sarcomere length, although the tension after the stretch was considerably greater than isometric. This suggests that an almost identical number of cross-bridges produce a greater tension after a slow stretch than during an isometric tetanus.


Force-velocity relations of the portal vein of hyperthyroid rats. The effect of thyroxine on the elementary process of contraction of vascular smooth muscle was tested on 38 hyperthyroid and in 38 control rats. Hyperthyroidism was induced by i.p. injections of triiodthyronine or 1-htyroxine for 2 weeks. By means of afterloaded isotonic contractions of the tetanized portal vein force velocity relations were calculated. There was a slight increase in both the extrapolated velocity of shortening at zero load [from 0.93 +/- 0.04 ML/s (control rats) to 1.03 +/- 0.04 ML/s (hyperthyroid rats; P less than 0.05)], and the peak force generation (from 14.8 +/- 0.4 mN to 16.1 +/- 0.4 mN; P less than 0.005). The maximum of mechanical power development at a distinct extent of afterload was augmented from 1.35 +/- 0.06 muW/ML to 1.68 +/- 0.06 muW/ML (P less than 0.005). The influence of thyroxine on the elementary process of contraction in vascular smooth muscle is discussed in connection with the much stronger effect of hyperthyroidism on cardiac muscle contraction.


Oxygen diffusivity in tumor tissue (DS-carcinosarcoma) under temperature conditions within the range of 20--40 degrees C. The O2 diffusion constants D and K of tumor tissue (DS-Carcinosarcoma in the rat kidney) were determined at temperatures of 20, 30, 37, and 40 degrees C. The following mean values were obtained for the conditions of 37 degrees C: D = 1.75-10(-5) cm2/s and K = 1.9-10(-5) mlO2/cm-min-atm. Within the range of 20-40 degrees C, temperature variations in tumor tissue cause changes in the O2 diffusion coefficient D of 2.0-2.5%/C and in the Krogh O2 diffusion constant K of 0.5-1.5%/C. The measured O2 diffusion constants for tumor tissue correspond to values of normal tissue with similar water content. This indicates that the insufficient O2 supply in DS-Carcinosarcoma is due not to unfavorable O2 diffusivity of the tumor tissue but rather to a decreased convective O2 transport and to insufficient capillarization. An analysis of O2 diffusion in DS-Carcinosarcoma tissue using the determined O2 diffusion constants lead to the result that, under the conditions of arterial normoxia and normocapnia, critical O2 supply conditions are to be expected when the intercapillary distance exceeds approximately 120 micrometer.


Activation of afferent cardiac sympathetic nerve fibers of the cat by pain producing substances and by noxious heat. Afferent discharges of 64 single units were recorded from the left cardiac sympathetic nerve of anesthetized cats. Mechano-sensitive terminals of the afferent fibers were localized in the extrapulmonary part of the pulmonary artery, left atrium, left ventricle and left pericardium, as determined by direct mechanical probing of the heart after death of the animals. Conduction velocity of the fibers ranged from 2.5 to 14.6 m/s. Excitation of these Adelta-fibers with mechanically excitable endings was produced by intravenous injections of acetylcholine, 5-hydroxytryptamine, bradykinin, histamine and veratridine, and/or by topical application of these agents to the receptor region. Noxious heat to the mechanically excitable field in the wall of the pulmonary artery and the left ventricle also activated their afferent fibers. These observations provide evidence for a certain number of afferent units in the cardiac sympathetic nerve with polymodal sensitivity. These afferent fibers can provide the spinal cord with information not only on mechanical changes in cardiac events, but also changes in the chemical environment of the cardiac nerve ending, possibly produced by myocardial ischemia.


Inward current of the rabbit sinoatrial node cell. Inward currents of the rabbit sinoatrial node cell were examined in voltage-clamp experiments using the two-microelectrode technique. A fast and slow inward current could be separated from each other. The slow inward current was blocked by Mn and D 600, but it was insensitive to TTX. On the contrary, the fast inward current was blocked by TTX, but not by Mn and D 600. Both the fast and slow inward currents disappeared on Na removal. The fast inward current system was fully inactivated by holding the membrane potential positive to -40 to -50 mV, while the slow inward current system was recorded with the holding potential up to -20 mV. The voltage dependence of the inactivation of the 2 inward current systems and their dependence on [Na]0 suggests that the rising phase of the spontaneous action potential in the S-A node cell is produced mainly by Na current through the slow inward current system.


Role of calcium in the release of noradrenaline induced by sodium deprivation from the guinea-pig vas deferens. The ability of Na-free media containing isotonic sucrose or equimolar choline chloride to increase the spontaneous release of noradrenaline from adrenergic nerve terminals and the dependence of the increase on extracellular calcium have been examined in the isolated guinea-pig vas deferens. The increase in noradrenaline output induced by the Na deprivation was gradual in contrast to the rapid response produced by excess potassium. The release of noradrenaline induced by the Na-free solution containing sucrose was not significantly reduced by the removal of calcium with or without EGTA in the first 1 h period, but was reversibly inhibited in the second 1 h period. On the other hand, the effect of the Na-free solution containing choline was significantly but reversibly decreased by exposure to the Ca-free media without any chelating agent in both periods of incubation. Lanthanum (0.25 mM) moderately inhibited the response induced by calcium reintroduction following exposure to the Ca-free, Na-free choline medium, but magnesium (10 or 20 mM) did not. It is suggested that Na deprivation may cause an increase in Ca efflux, a decrease in Ca efflux or a release of intracellulary stored calcium resulting in an increase in the level of intracellular free calcium which then stimulates the output of noradrenaline.


Studies on the binding characteristics of antimycin A and albumin in relation to the inhibitor activity of the complex on rat proximal tubular sodium transport. As reported previously [11] Antimycin A is effective in inhibiting sodium transport of the proximal tubule only when applied to the luminal side and in the presence of albumin. Therefore the interaction of Antimycin A with albumin was examined with the technique of equilibrium dialysis. It was found that Antimycin A was bound to albumin at five sites with a dissociation constant of 2.5 X 10(-6) M. This finding suggests that Antimycin A is taken up by the tubular cell as an Antimycin A/albumin complex via pinocytosis. In the pinocytotic vesicle this complex probably dissociates, and free Antimycin A is released into the cytoplasm where it can reach it's sites of action in the mitochondria and at the plasma membrane. This uptake mechanism might provide a general method to incorporate substances into the cell which do not penetrate the plasma membrane.


Effect of exercise on systemic blood pressure and heart rate in horses. Carotid loops were prepared in 3 horses several months prior to the experiments. Systemic blood pressure was recorded at rest and during exercise by insertion of a plastic cannula into the carotid artery. The pressure transducer was fixed at the neck of the animal. The blood pressure signal was transmitted by telemetry. When the horses were standing under the rider, the following results were obtained: heart rate 38 +/- 5 beats-min-1, systolic pressure 115 +/- 15, disstolic pressure 83 +/- 10, mean pressure 97 +/- 12, and pulse pressure 32 +/- 9 mm Hg. During steady gallop at a mean speed of 548 +/- 90 m-min-1, heart rate rose to 184 +/- 23 beats-min-1, systolic pressure to 205 +/- 23, diastolic pressure to 116 +/- 12, mean pressure to 160 +/- 20 and pulse pressure to 89 +/- 19 mm Hg. These values remained stable throughout the exercise period of 5-6 min. When the horses were exercised at stepwise increasing speed from walk through trot to gallop, both the mean arterial blood pressure and the pulse pressure rose in proportion to the running speed.


Nucleic acid and protein content of rat cecum mucosa in dietary adaptation-growth by cellular hyperplasia. The deoxyribonucleic acid (DNA), ribonucleic acid (RNA) and protein content of the isolated cecum mucosa was determined in control rats and rats with adaptive cecum growth induced by dietary polyethylene glycol. Feeding the polymer resulted in an increase in the total amount of mucosal nucleic acids and protein which was statistically significant after 2-4 days and reached twice the control value after 2 months. During growth, the protein/DNA, protein/RNA and RNA/DNA ratios remained unaltered, indicating hyperplasia and not hypertrophy of the epithelial cells. The water content of the mucosa increased from 75 to 81% during adaptation, possibly related to the stimulation in active sodium transport reported earlier. Recalculation of parallel Na-K-ATPase data on the basis of DNA demonstrated a doubling of Na-K-ATPase activity per mg DNA in the fully adapted cell.


[Chronic renal failure following intoxication by Cortinarius orellanus fries type mushrooms. Two cases of familial intoxication (author's transl)]. Poisoning from Cortina orellanus is to be observed when favourable weather conditions allow the spread of the Cortina in the mountains. A failure to recognize toxicity from this mushroom and a few related species explains the occurrence of genuine epiemics of poisoning, such as has been observed in Poland. Two case histories reported by the authors illustrate that such cases of poisoning occur in France, where the most dangerous of the Cortina, Cortina orellanus, although not widespread, is also not rare. The clinical picture is notable by virtue of the long incubation period and predominantly renal involvement, which is paramount in diagnosis. The condition is initially one of an acute interstitial tubular nephritis which is likely to develop later into a chronic condition.


The presence of ovalbumin mRNA coding sequences in multiple restriction fragments of chicken DNA. Chicken DNA has been digested with restriction enzymes and the size distribution of the DNA fragments containing ovalbumin specific sequences has been examined after separation of the fragments on agarose gels and transfer to nitrocellulose sheets. Hybridisation with terminally 32P-labelled ovalbumin mRNA fragments or with RNA populations transcribed from the DNA of a hybrid plasmid containing ovalbumin sequences was used to locate the DNA fragments coding for ovalbumin. Digestion with enzymes which do not cut within the portion of the ovalbumin gene synthesised from ovalbumin messenger RNA in vitro has shown the presence of several defined fragments carrying ovalbumin specific sequences. Possible explanations of these observations are discussed.


Effect of propranolol on glucose tolerance in hyperthyroidism. Glucose tolerance tests were performed on six hyperthyroid patients in the morning and afternoon before, and at the end of treatment with propranolol for 2 weeks. All six subjects showed normal glucose tolerance with a normal diurnal rhythm before treatment and this remained normal in five during treatment with propranolol, while one patient developed abnormal glucose tolerance and the diurnal variation was abolished. No change in insulin levels was noted in any of the patients either before or during treatment. Propranolol may cause deterioration in glucose tolerance in some subjects with hyperthyroidism and this possibility should not be overlooked.


Cycloheximide produces adult-like retention deficits of prior learning in infant mice. Utilizing a dosage of cycloheximide which was found to inhibit cerebral protein synthesis by almost 90% after injection, separate groups of 13-day-old mice received either cycloheximide or saline followed by 0 (control), 15, or 25 training trials in a discriminated shock-escape T-maze. Twenty-four hr later, each mouse was treated with cycloheximide or saline and tested for retention by an additional 25 trails in the T-maze. As reflected by correct choice-point turns, the results suggest that whereas salinetreated mice demonstrated reliable retention of prior learning, cycloheximide treated mice exhibited memory impairment; cycloheximide per se had no effect on performance during either original training or retest. A final experiment indicated that this memory impairment was not due to cycloheximide's general debilitating side effects at the time of retention testing. Taken together, these data suggest that protein synthesis inhibition during training impaired consolidation and/or retrieval processes involved in memory. The biochemical and behavioral effects following cycloheximide injection in 13-14-day-old mice in the present study parallel those reported with adult animals and lend indirect support to the hypothesis that the 24-hr memory capacity exhibited by these young mice reflects the early functioning of those processes involved in adult long-term memory.


Concurrent intracranial self-stimulation and amphetamine self-administration in rats. In a two-lever testing chamber, rats had concurrent access to intravenous amphetamine and brain stimulation reinforcers. Responding for each reinforcer was generally increased above baseline rates taken when only one reinforcer was available. Amphetamine stereotypy was observed, but did not interfere with rapid lever-pressing for brain stimulation.


Derivatives of 3.3-spiro-bi-5-methyl-5'-alkylaminomethyl-tetrahydrofuranone-2 as potential neurotropic agents. II. Dilactone of gamma-bromo-beta-hydroxypropyl-beta-hydroxypropyl malonic acid 2 was prepared. By condensation with secondary or primary amines the tertiary amines 3--7. resp. the secondary amines 8--10 were obtained. The reaction of 2 with tertiary amines gave the corresponding ammonium salts 11--12. All the compounds are subjected to pharmacological examination.


Ethanol dependence as a determinant of fluid preference. Rats made dependent on ethanol by a schedule-induced polydipsia procedure preferred 5% ethanol to an increasing concentration of dextrose solution to a greater extent than animals on a non-dependent, non-polydipsic procedure which allowed an equivalent opportunity to drink ethanol, confirming a previous study. Two corresponding groups of animals drinking isotonic (0.9%) NaCl rather than 5% ethanol behaved similarly to the latter group, changing to a dextrose preference at a lower dextrose concentration than the ethanol polydipsic group. Therefore, neither the intermittent food regimen (polydipsia-generating procedure) in itself, nor a history of isotonic saline polydipsia biased fluid preference against dextrose solution choices. The enhanced preference for ethanol over dextrose solutions shown by the ethanol polydipsic group can be attributed to physical dependence rather than regiment produced artifacts.