Development of trophoblast and placenta of the mouse. A reinvestigation with regard to the in vitro culture of mouse trophoblast and placenta. At 5 days post conceptionem (p.c.) shortly after implantation, giant cell transformation starts at the abembryonic pole of the blastocyst, spreading over the mural trophoblast; 1 day later, the first ectoplacental giant cells appear at the base of the fast growing ectoplacental cone (derived from the polar trophoblast). Giant cell transformation expands over it periphery. Thus, by the 8th day p.c., the conceptus is separated from the maternal tissue by a continuous layer of giant cells, variable in thickness. Giant cells reach their greatest size by 10 days p.c. in the mural tophoblast and by 12 days p.c. in the chorioallantoic placenta. They are probably no longer formed after that stage. Around the 8th day p.c., the allantois reaches contact with the ectoplacental cone, which develops into the chorioallantoic (definitive) placenta. At 9 days p.c., its four zones can already be discriminated: chorionic plate, labyrinth, junctional zone (trophospongium), and zone of giant cells, respectively. Within the next day, the chorioallantoic placental circulation is established. The yolk sac placental circulation is established by the 9th day p.c. The villi of the proximal layer of the yolk sac increase in size and number, and their capillary network becomes more dense until the 12th to 14th day p.c. This provides evidence that the yolk sac placenta exerts its function--to a certain extent--beyond the establishment of the definitive placenta. Around the 14th day p.c., the placental labyrinth reaches its definitive features. Fetal capillaries in the labyrinth, branching from unbilical blood vessels within the septa of connective tissue are surrounded by trophoblast cells. They form a dense vascular network bathing in maternal blood. The structures of the placental zones remain almost the same during further development, the borders becoming sometimes little blurred. Adjacent to the chorionic plate, subchorionic clefts appear at the 14th day p.c. These clefts become confluent to form the intraplacental space, regularly communicating with the yolk sac cavity. At the end of gestation (19th day p.c.) there is a considerable amount of eosinophilic material ('fibrinoid') between the zone of giant cells and the decidua, probably produced by the giant cells.


[Origin and function of globular leukocytes]. The morphology of the globule leukocyte (GL) has been investigated by light and electron microscopy in normal (decidua of mouse placenta) and pathologically altered tissues (tumors of epidermis and mucous membrane in men). The results make it probable that the GL my originate from mast cells as well as from eosinophil granulocytes. The high content of basic proteins within the globules of GL has been interpreted as a nutritive function by this cell type to the surrounding tissue.


The innervation of the umbilical cord of the rat. A histochemical study. The umbilical cords of 21 days old rat foetuses were investigated using histochemical methods for acetylcholinesterase and catecholamines. An AChE positive nerve plexus is situated only around the vitelline vessels. At regular intervals the bundles of this plexus exhibit small ganglia. These ganglia are made up of nerve cells, which are AChE positive and show formaldehyde induced fluorescence, thus indicating an adrenergic nature of these cells. No innervation could be found in the allantoic part of the umbilical cord.


Echocardiographic features of combined membranous subaortic stenosis and acquired calcific aortic valvulopathy. The M-mode echocardiographic features of aortic valve structure and motion in a 45-year-old male with combined congenital subaortic diaphragm and acquired deformity of the aortic valve are described. Clinical, hemodynamic, and angiographic studies suggested calcific aortic valve disease with stenosis and insufficiency, but the additional presence of a subaortic diaphragm was not appreciated. Cardiac ultrasonography demonstrated multiple, central diastolic aortic valve cusp echoes consistent with a thickened, calcified, tricuspid aortic valve. Despite calcification of the cusps, however, enough systolic cusp excursion remained to demonstrate an early systolic, rapid movement toward closure of the right coronary cusp-a finding suggestive of fixed subvalvular obstruction. Surgery confirmed a discrete subaortic diaphragm and a tricuspid, thichened, mildly calcified aortic valve with fusion of the cusp commissures at their origins and rolling back of the cusp edges. The value of echocardiography in the evaluation of the left ventricular outflow tract and aortic valve is emphasized.


Trichinosis in bears of western and northcentral United States. Black bears (Ursus americanus) from 10 states were examined for Trichinella spiralis during the 1970-1975 period. Trichinae were found in 14 (3.1%) of 454 bears examined. Infected bears were found from California, seven (13.2%) of 54; Idaho, one (2.3%) of 44; and Wisconsin, six (3.8%) of 163. Trichina counts per gram of tissue obtained by the artificial digestion-Baermann method ranged from 1390 to 0.02. Six California bears and one from Wisconsin contained more than one trichina per gram, a level considered capable of inducing clinical trichinosis in man.


"Prostatic acid phosphatase?" A comparison of acid phosphatase activities in epithelial cells, granulocytes, monocytes, lymphocytes, and platelets purified by velocity sedimentation in isokinetic gradients of Ficoll in tissue culture medium. Numerous investigators have found several substrates and inhibitors to be particularly suited for the demonstration of acid phosphatase of prostatic origin. There has been much controversy over the specificity or lack of specificity of several substrates and inhibitors. We have investigated acid phosphatase activities obtained from several kinds of purified cells. None of the substrates or inhibitors which we studied permitted us to discriminate "prostatic" acid phosphatase from acid phosphatase activities obtained from other kinds of cells.


Rape: a sexual deviation.

Rape: a sexual deviation. Clinical work with offenders and victims is cited to support the concept of rape as a sexual deviation, as well as a sexual offense. It is suggested that rape is directed toward the sexual expression and gratification of needs that are not basically sexual, and that it represents a developmental crisis for the offender, which in turn triggers a situational crisis for the victim.


Influence of personality attributes on abortion experiences. Questionnaire responses of women who underwent legal abortions reveal a range of complex emotional reactions. This study suggests that both the resolution of negative feelings post-abortion and the motivation to use contraception are related to individual personality characteristics. Implications are offered for therapeutic intervention and for contraceptive counseling with young, unmarried women.


Trematode eggs in the peritoneal cavity of man in Honduras. Surgical repair of an inguinal hernia in a 19-year-old man in Honduras revealed massive numbers of small granulomata containing trematode eggs on the omentum and other peritoneal surfaces. The eggs resembled those of Achillurbainia recondita Travassos, 1942, a species found in the maxillary sinuses of the opossum, Didelphis marsupialis. Species of Achillurbainis (syn. Poikilorchis Fain and Vandepitte, 1957) have been reported in retroauricular cysts or abscesses in residents of West Africa and Southeast Asia.


[Influence of ionizing radiation on the formation of chondrocranium in merino-sheep. II (author's transl)]. The influence of ionizing radiation on the formation of chondrocranium in merino-sheep was studied on the 28th, 29th and 30th day of fetus development following the exposition of 250 R. On the 28th day, after the radiation, delayed growth of fetus head was evoked. From the whole of chondrocranium, it was found that only lamina basalis and membranous parts of nasal and optical capsules were developed. On the 29th and 30th day after radiation, a prolonged period of development of chondrocranium was noticed. The greatest changes were seen in capsula nasalis and otica. In all the three groups some abnormal findings were found on the bases of the eyes.


The morphodynamic patterns of the paraventricular nucleus in rats under experimental conditions. I. Cytoarchitectonic studies. An elaboration of the cytoarchitectonics of the PVN and the topography of the neurosecretory cells scattered in the anterior hypothalamus in rats subjected to various experiments is presented. The most distinct and complete patterns were obtained from rats with experimental occlusion of the small intestine, in the early post-laparotomy period and after withdrawal of food and water for periods of 6, 12, 24 hours. A hitherto undiscovered group of neurosecretory cells was found in the stria terminalis and single cells of that type near the anterior commissura.


Histomorphological and morphometric changes in the spleen of ageing goats. The spleens of twenty four goats of both the sex and the females at various phases of estrus cycle aged from 20 days to over five years have been studied for their varying histomorphology with age. The morphometric studies included the computation of percent white pulp, percent red pulp, percent trabecular tissue, size of white pulp follicles and number of white pulp follicles per cm2 area. Agewise and sexual and cyclic changes in these parameters have been presented with the help of statistically analysed data by applying multiple range test and computing the correlation coefficients and regression equations. The interrelationship between different parameters have also been considered. The results have been discussed in light of above with graphic presentation including the scatter diagrams and the regression lines.


The causation of two scent-marking behaviour patterns in female hamsters (Mesocricetus auratus). Social, olfactory, and oestrous cycle influences on the frequency of flank-marking and vaginal marking were studied in female hamsters. Vaginal marking was more frequent in the presence of males or their odours than females or their odours. Vaginal marking frequency was greatest the day before oestrus (day 4), intermediate on days 2 and 3, and zero on the oestrous day. The probable sexual advertisement functions of vaginal marking were discussed. In contrast, flank-marking by females was stimulated more by other female odours than by male odours. The frequency of flank-marking was elevated by agonistic encounters and was reduced by sexual encounters. Flank-marking seems to have functions related to functions of aggression among hamsters.


Space utilization of Peromyscus: social and spatial factors. Space utilization of Peromyscus maniculatus bairdi and P. leucopus noveboracensis was studied in laboratory enclosures. After 1 week, individuals were given access to twice as much space for an additional week. The new rea was either vacant or was occupied by a similarly experienced conspecific of the same or opposite sex. Patterns of space utilization and reaction to the doubling of space were species specific. Each species displayed specific patterns of social behaviour and only P. maniculatus reproduced. Both species exhibited similar patterns of space utilization in the presence of another individual. Social regulation of space utilization did not result from territorial defence of mutually exclusive areas, but from the initial mutual avoidance of two individuals.


Experimental and naturally occurring transplacental transmission of canine distemper virus. In the present study, 2 different effects of experimentally induced infection with virulent canine distemper virus (CDV) on pregnant CDV-susceptible dogs were studied. In 1 bitch, abortion occurred 7 days after viral inoculation and there was no evidence of fetal infection. Another bitch had subclinical infection and delivered 3 CDV-infected pups. Sequential clinical, immunologic, and virologic studies of a litter of gnotobiotic pups (3rd bitch) that were congenitally infected with CDV demonstrated the heightened susceptibility to CDV in the neonatal period. The data presented add canine distemper to the list of transplacental infectious diseases in the canine species.


Alkaline phosphatase, leucine aminopeptidase, and alanine aminotransferase activities with obstructive and toxic hepatic disease in cats. The activities of serum alkaline phosphatase (serum ALP), leucine aminopeptidase (serum LAP), and alanine aminotransferase (serum ALT) were determined in 15 cats before and after treatment by 3 methods: common bile duct occlusion, left hepatic duct(s) occlusion, and carbon tetrachloride administration. Significant increases in serum ALP, LAP, and ALT activities occurred in all cats in the 3 groups. Sustained mean increases of ninefold in ALP and 13-fold in LAP occurred in the cats with common bile duct occlusion. Lesser mean increases of these enzymes (fourfold) occurred in the cats with partial biliary occlusion. Transient mean increases (100-fold) in ALT occurred in the carbon tetrachloride-treated cats. Urine ALP excretion was measured in 3 cats with common bile duct occlusion. There was no significant difference between rates of urine ALP excretion before and after common bile duct occlusion. Specific ALP activities of hepatic extracts from normal cats and biliary-obstructed cats were compared. Mean specific activity was onefold higher in liver from cats with common bile duct occlusion of 21 days' duration. The findings in the present studies were interpreted to indicate that serum ALP and LAP are useful to detect biliary occlusive disease in cats and, in conjunction with serum ALT, may be used to differentiate primary hepatodegenerative disease  and biliary occlusive disease.


[Effect of antibiotics on the structure and function of the ovaries]. Streptomycin, penicillin, levomycetin, florimycin, cycloserin, erythromycin, sekazin, novobiocin, tetracyclines and neomycins had a stimulating effect on the sexual organs of the tests animals during the 1st decade of their administration. With long-term use some of the drugs induced dystrophic changes in the ovary. The signs of stimulation in the ovary were evident from increased activity of the oxidation-reduction enzymes in the inner membrane of the follicles and the interstitial cells of the stroma, as well as in an increase in the number of the maturating follicles. With long-term long use of erythromycin, sekasin or novobiocin, disappearance of the primordial follicles, an increase in the number of the atretic follicles and death of the ovocytes in the mature follicles were observed in the ovary.


Valve replacement with the Starr-Edwards and Hancock prostheses: comparative analysis of late morbidity and mortality. Although the Starr-Edwards caged-ball valve remains a standard of comparison for more recently introduced prostheses, a substantial incidence of thromboembolic and hemorrhagic complications prompted our evaluation of the Hancock glutaraldehyde-fixed porcine xenograft. We have compared the results of 435 aortic valve replacements using the Starr- Edwards valve (SE-AVR), 515 mitral valve replacements (SE-MVR), and 121 double-valve replacements (SE-AVRMVR) with 251 aortic valve replacements using the xenograft aortic valve (X-AVR), 338 mitral valve replacements (X-MVR), and 88 double-valve replacements (X-AVR-MVR). The Starr- Edwards valves were used during the period 1963 through 1973 and the xenograft valves between 1971 and 1976. No significant differences in patient age, sex, or preoperative hemodynamic data were noted between comparable groups. All patients with Starr-Edwards valves received long-term anticoagulation while anticoagulants were used only for specific indications in patients with xenograft valves. Total follow up was 3944 patient years for the Starr-Edwards patients and 947 patient years for the xenograft patients. Hospital mortality was not significantly different for comparable groups: SE-AVR 6.9% vs. X-AVR 6.4%, SE-MVR 9.7% vs X-MVR 8.6%, and SE-AVR-MVR 7.5% vs. X-AVR-MVR 10.2%. Linearized mortality and morbidity data expressed as percent per patient- year are tabulated below. Pairs which differ significantly (p < .05) are italicized.


Ophiocordin, an antifungal antibiotic of Cordyceps ophioglossoides. An unknown antibiotic, ophiocordin, C21H22N2O8, MW: 430, was isolated from submerged cultures of Cordyceps ophioglossoides, strain TU 276, grown in a glycerol soybean meal medium at 27 degrees C. The antibiotic was extracted from acidified culture fluids with n-butanol and purified by subsequent column chromatography on DEAE-Sephadex and cellulose. Studies including nuclear magnetic resonance and mass spectrometry resulted in proposals of partial structures of the molecule. Inhibition by ophiocordin could be demonstrated for a small number of fungi belonging to different taxonomic groups. Bacteria were not inhibited. The antifungal effect was antagonized by ammonia and nitrate ions and by certain amino acids.


Sophorose metabolism and cellulase induction in Trichoderma. The cellulase inducer sophorose was rapidly catabolized to CO2 and H2O by Trichoderma: only small amounts were used to induce the synthesis of cellulase. 3H-sophorose uptake began after a lag of 1 h and its half-life in the medium was less than 5 h. Cellulase activity in the medium did not increase till 6 h after the addition of sophorose and reached a half maximum value at 14 h. The presence of free sophorose in the medium was required for continuous cellulase production. Several small sophorose addition induced much more cellulase than an equivalent single dose. These results are attributed to two pathways of sophorose utilization, a catabolic pathway that has a high capacity but low affinity for sophorose and an inductive pathway having a lower capacity but higher affinity for sophorose.


Influence of specific growth limitation and dilution rate on the phosphorylation efficiency and cytochrome content of mitochondria of Candida utilis NCYC 321. With Candida utilis cells that had been removed directly from a 61 chemostat culture, in steady state, well-coupled mitochondria generally could be isolated. This requires a modified snail-gut enzyme procedure that allowed the total processing time to be decreased to 3 h, or less. Examination of these mitochondria in an oxygraph showed the presence of 3 sites of energy conservation when the cells were grown at various dilution rates between 0.1 and 0.45 h-1 in environments that were, successively, glucose-, ammonia-, magnesium-, phosphate- and sulphate-limited. Potassium-limited cells also apparently possessed 3 sites of oxidative phosphorylation when growing at dilution rates greater than 0.2 h-1, but only 2 sites when growing at lower dilution rates. Analysis of cytochrome spectra obtained with these intact mitochondria revealed large quantitative (but not qualitative) differences, depending on the environmental conditions under which the yeast had been cultured. In particular, comparison of the ratio of cytochrome b to cytochrome a showed a pattern of change with dilution rate in mitochondria from potassium-limited cells that was distinctly different from those evident in mitochondria from cells that had been limited in their growth by the availability of other nutrients.


Cell wall formation in zoospores of Allomyces arbuscula. III. Carbohydrate composition of cell walls during development from meiospores to hyphae. Carbohydrate composition was determined in isolated cell walls of meiospores of Allomyces arbuscula after incubation for 15 min (encysted meiospores: cysts), 150 min (germlings: cysts + rhizoids) and 24 h (cysts + rhizoids + hyphae). The principal constituent in all cell wall samples is chitin, accounting for about 75% of the recovered carbohydrates. In addition, cell walls of all stages examined contain polysaccharides which release galactose, glucose, mannose, arabinose, xylose, fucose, and rhamnose on acid hydrolysis. While different developmental stages show minor quantitative changes in chitin, the ratio of galactose to glucose decreases sharply during differentiation of ungerminated cysts into germlings with rhizoids and hyphae. The increase in glucose is accompanied by a decrease in the amount of xylose and/or fucose and of galactose.


An alkali-soluble polysaccharide from the cell walls of Coprinus lagopus. An alkali-soluble polysaccharide was isolated from the purified mycelial walls of Coprinus lagopus. The hydrolysis products, optical rotation, and infrared spectrum indicate a beta-glucan. Hydrolysis of the glucan after permethylation gave only 2,3,4,6-tetra-, 2,4,6-tri-, and 2,4-di-O-methyl-D-glucose. These methylated sugars and their relative quantities reveal that the glucan is a polysaccharide containing beta-1,3-linked glucose units with about 14% of the sugars having 1,6-linked branch points. Partial hydrolysis of the product derived from Smith degradation of the glucan released laminaribiose and gentiobiose suggesting that the branches are generally longer than a single glucose unit.


Feedback inhibition of IgM memory cells by high antigen dose. The ability of high and low antigen doses (SRBC) to recruit IgM memory cells has been compared in several strains of mice. In intact animals priming with low doses was more efficient than priming with high doses. If, however, mice of different strains were X-irradiated two days after priming and repopulated, regardless of whether syngeneic or allogeneic splenocytes were used, they fell into two categories--those in which more memory cells were found after low antigen priming and those in which the reverse was true (Swiss mice). Two possible explanations are offered to explain these interstrain differences--antibody mediated suppression, and generation of suppressor T cells. Our data favor the latter, and we assume that suppressor T cells appear at different intervals after priming in different strains of mice, and that these cells are radioresistant.


The phosphoglucomutase group system of human erythrocytes. Frequency of PGM1 phenotypes and their corresponding genes in the Polish population was determined in two samples: in 760 adults and 240 children. The frequency of the PGM1 1 gene in adults was 0-748, and in children 0-765. A statistically significant deficiency of heterozygotes was found in adults. Heredity of the phosphoglucomutase system was studied in 52 families with 162 children, in 223 mother-child pairs, and in 73 twin pairs of the same sex. In all groups, phenotypes of PGM1 agreed with the hypothesis of heredity, assuming a pair of alleles of the PGM1 system. In the parent combinations PGM1 1-1 X PGM1 2-1 a deficiency of PGM1 2-1 children was found. Analysis of linkage of genes of the PGM1 system with eight group systems provided data indicating a possibility of linkage of the PGM1 system with the Rh system. The theoretical usefulness of the PGM1 system in paternity investigations in the Polish population was estimated to be 14-24%.


Biological and biophysical characteristics of mouse adenovirus, strain FL. Mouse adenovirus, strain FL (MAV), is best propagated in primary rather than secondary cultures of mouse kidney cells. Virus release from the cells into the medium is fairly efficient; the average yield per cell is about 1000 TCID50. Morphological and other biophysical characteristics are those of a typical adenovirus: the icosahedral shell with a diameter of 74 nm, fiber projections of 29 nm length, its localization inside the infected kidney cells, a buoyant density in CsC1 of 1.34 g/ml, inhibition of multiplication by inhibitors of DNA synthesis. The thermostability is higher than that of human adenoviruses, whereas, in contrast to these, MAV is inactivated to a great extent by trypsin. Complete or incomplete hemagglutinin or toxin-like activity were not detected. The virus shows no cross-neutralization with human adenoviruses and a one-side cross-reaction with another murine adenovirus strain K-87. Soluble complement-fixing antigen of MAX exhibits a sedimentation rate of 12S identical to the hexon component of human adenoviruses; both antigens show a partial antigenic relationship in tests with appropriate antisera.


Improved method for the purification of hog cholera virus grown in tissue culture. A method for purification of Hog Cholera Virus (HCV) is presented. Cell-associated virus was extracted from PK15 cells 17 hours p.i. by fluorocarbon treatment. The virus was concentrated by polyethylene glycol precipitation and partially purified by pelleting on a fluorocarbon cushion. Final purification was achieved by rate zonal centrifugation in a 7--35 per cent sucrose gradient. This procedure permits a recovery of approximately 40 per cent of viral infectivity. A specific infectivity of about 2 X 10(7) PFU/microgram of protein was achieved. An apparent density d=1.13 g/ml in sucrose and a Sw20 value of about 150 was determined for purified HC virions.


The effect of chemical treatments of albumin and orosomucoid on rate of clearance from the rat bloodstream and rate of pinocytic capture of rat yolk sac cultured in vitro. Portions of a 125I-iodinated bovine serum albumin preparation were exposed to freezing, acetic acid (pH 3.5, 3.0 or 2.5), urea or formaldehyde, and the effect of these treatments on the rates of pinocytic uptake by yolk sacs from 17.5-day-pregnant rats cultured in vitro and of clearance from the rat bloodstream were studied. Uptake of albumin by the yolk sac was followed by rapid release of [125I]iodo-L-tyrosine into the culture medium. Similarly clearance of albumin in vivo was accompanied by the appearance of trichloroacetic acid-soluble radioactivity in the bloodstream. In both systems the rates of uptake of modified albumin preparations formed a series: formaldehyde or urea greater than acetic acid greater than freezing. The increased rates of uptake of modified albumin preparations could not be ascribed to the formation of aggregates nor, in the yolk-sac system, to an increase in the rate of pinosome formation. It is concluded that the various treatments to which the albumin was subjected increase to varying degrees the affinity of the albumin molecule for binding sites on that region of the plasma membrane from which pinocytic vesicles are formed. Some comparable experiments with native and desialylated human orosomucoid indicate that the rat yolk-sac epithelial cells do not possess the recognition system for uptake of asialoglycoproteins that exists on the surface of hepatic parenchymal cells.


Methods of recording fetal movement.

Methods of recording fetal movement. Patient, observer and instrumental methods for recording fetal movements are described. Significant correlations between both patient and observer, and patient and instrumental fetal movement counts, were found. Significant diurnal, daily and weekly variation in fetal movement counts occurred which need to be considered in the interpretation of the frequency of fetal movements as an indication of fetal welfare.


Conformation of DNA modified with a dihydrodiol epoxide derivative of benzo[a]pyrene. The conformation of calf thymus DNA modified by reaction with (+/-)-7beta,8alpha-dihydroxy-9alpha,10alpha-epoxy7,8,9,10-tetrahydrobenzo[a]pyrene, which binds covalently mainly to the 2-amino group of guanosine residues, was studied. With samples in which 1.5 or 2.2% of the bases were modified, there was a slight decrease in Tm during heat denaturation and a slight increase in susceptibility to the single strand specific nuclease S1. In a DNA sample in which 4.5% of the bases were modified, there was an appreciable decrease in Tm and a marked increase in susceptibility to S1 nuclease. The kinetics of the reaction of the modified DNAs with formaldehyde provided evidence for locally destabilized regions ranging from 1 to 7 base plates, depending on the extent of modification. Alkaline and neutral sucrose gradient analyses revealed no evidence for strand breakage in the 1.5 and 2.2% modified samples, although single-strand breaks were found in the 4.5% modified samples. Taken together, these results suggest that DNA molecules containing a covalently bound benzo[a]pyrene derivative have an altered conformation characterized by small localized regions which are destabilized and easily denatured. The conformational changes associated with the covalent binding of the benzo[a]pyrene derivative to native DNA appear to be different from, and less marked, than those associated with the covalent binding of N-2-acetylaminofluorene to native DNA.


Mutants of CHO cells resistant to the protein synthesis inhibitors, cryptopleurine and tylocrebrine: genetic and biochemical evidence for common site of action of emetine, cryptopleurine, tylocrebine, and tubulosine. Stable mutants resistant to the protein synthesis inhibitors cryptopleurine and tylocrebine can be isolated in Chinese hamster ovary (CHO) cells, in a single step. The frequency of occurrence of cryptopleurine (CryR) and tylocrebrine (TylR) resistant mutants in normal and mutagenized cell populations is similar to that observed for emetine resistant (EmtR) mutants. The CryR, TylR, and EmtR mutants exhibit strikingly similar cross-resistance to the three drugs used for selection, to tubulosine and also to two emetine derivatives cephaeline and dehydroemetine, based on assays of in vivo cytotoxicity and on assays of protein synthesis in cell-free extracts. The identity of cross-resistance patterns of the CryR, TylR, and EmtR mutants indicates that the resistance to all these compounds results from the same primary lesion, which in the case of EmtR cells has been shown to affect the 40S ribosomal subunit. This conclusion is strongly supported by the failure of EmtR, TylR, and CryR mutants to complement each other in somatic cell hybrids. Based on these results it is suggested that the above group of compounds possesses common structural determinants which are responsible for their activity. The above mutants, however, do not show any cross-resistance to other inhibitors of protein synthesis such as cycloheximide, trichodermin, anisomycin, pactamycin, and sparsomycin, either in vivo or in vitro, indicating that the site of action of these inhibitors is different from that of the emetine-like compounds.


Structural transitions of calf thymus DNA in concentrated LiCl solutions. The solubility, sedimentation, circular dichroism, and absorption spectral characteristics of calf thymus DNA have been examined in concentrated solutions of LiCl (6-13 m) at 25 to 27 degree C. At all concentrations of LiCl, the DNA is base stacked and exhibits normal hypochromicty, At the upper end of this range of LiCl concentrations, DNA aggregates and ultimately precipitates completely from solution between 13 and 14 m LiCl. This aggregation process is dependent on concentration, base composition, and molecular weight of DNA. The sedimentation velocity data taken together with the absorbance spectral data suggest that the aggregation process leading to the formaiton of large structures beings at approximately equal to 9 m. Prior to the onset of aggregation, the circular dichroism (CD) spectra can be adequately fitted by a linear combination of contributions of the B, C, and A forms of DNA (Hanlon, S., Brudno, S., Wu, T. T., and Wolf, B. (1975), Biochemistry 14, 1648). Above 9 m LiCl, both factor analysis and a primitive version of matrix rank order analysis indicate that at least one additional spectral component is required to account for the observed CD spectra above 260 nm. The general shape of this additional component or distortion resembles the psi form of DNA.


Kinetic properties of carboxypeptidase B in solutions and crystals. The correlation of the structure of crystalline enzymes with their activities in solution assumes that the catalytic properties are identical in the two physical states. The present data demonstrate that in bovine carboxypeptidase B they differ significantly. Normal Michaelis-Menten kinetics characterize the hydrolysis of several esters and peptide substrates in both physical states. Crystallization reduces kcat 16 to 320-fold, while it affects KM variably and less dramatically. Small molecules inhibit catalytic activity both in solution and in crystals, but the carboxypeptidase inhibitor from potatoes (molecular weight 4200) does no inhibit the crystals. The activities of bovine carboxypeptidases A and B toward identical substrates are more similar in their crystals than in their solutions. This suggests that, over and above the structural dissimilarity of their crystals, conformational differences may additionally determine the activities of the two enzymes in solution. The findings demonstrate that the catalytic properties of carboxypeptidase B depend critically on its physical state.


Stimulation of lymphocytes by concanavalin A. Temperature-dependent effect of fatty acid replacements. The membrane fatty acyl composition of lymphocytes was altered by growth in lipid-depleted serum containing fatty acid supplements, as well as avidin to block endogenous synthesis of fatty acids. Under these growth conditions over 50% of the total fatty acid in membrane phospholipid were derived from the added fatty acid. Enrichment of lymphocyte membranes with oleate (cis C18:1) or elaidate (trans C18:1) shifted the optimum temperature for mitogenic stimulation by concanavalin A as measured by [3H]thymidine incorporation. These results suggest that the fluidity of the membrane lipid phase plays a role in the process of lymphocyte stimulation by lectins.


Spontaneous reactivation of acetylcholinesterase following organophosphate inhibition. II. Characterization of the reactivating components. Repeated cycles of inhibition by a variety of organophosphates followed by spontaneous reactivation reveal a component of electric eel acetylcholinesterase (acetylcholine hydrolase, EC 3.1.1.7) which preferentially reactivates. That the observed enzymatic activity truly resides in acetylcholinesterase is indicated by its sensitivity to a specific inhibitor and by molecular weights for subunits and native enzyme which are approximately the same as those for the major fraction of enzymatic activity which behaves in the classical manner. The Km values for phenyl acetate of the two components are similar but the rate constant for covalent bond formation, k2, with isopropyl m-nitrophenyl methylphosphonate is greatly reduced in the spontaneously reactivating species. The molecular basis for these observations is discussed.


Physical and kinetic distinction of two ornithine decarboxylase forms in Physarum. Two forms of ornithine decarboxylase (L-ornithine carboxy-lyase, EC 4.1.1.17) can be isolated from crude plasmodial homogenates of Physarum polycephalum. Both forms catalyze the stoichiometric production of putrescine and CO2 from ornithine, yet they are distinguished by (a) a large difference in their affinity for coenzyme (apparent Km values of 0.13 and 33 muM); (b) a differential stability to extended dialysis of crude homogenates at 4 degrees C; and (c) the tendency of the low affinity form to polymerize when suspended in low ionic strength borate and phosphate buffers. These forms appear to be alternate states of a basic catalytic subunit in that (a) they both demonstrate monomer and dimer molecular forms of 80 000 and 160 000 daltons, respectively, depending on the buffer content; (b) they coelute from DEAE-Cellulose ion-exchange columns; and (c) they vary in activity in approximately equivalent yet opposite directions in response to factors which alter this organism's growth or metabolism. These data suggest that ornithine decarboxylase activity may be modulated by the control of the transition of this enzyme between the active and the relatively less active form.


Substrate-inhibiton by acetyl-CoA in the condensation reaction between oxaloacetate and acetyl-CoA catalyzed by citrate synthase from pig heart. Deviations from Michealis-Menten kinetics in the pig-heart citrate synthase (citrate-oxaloacetate-lyase(pro-3S-CH2-COO-leads to acetyl-CoA), EC 4.1.3.7) system have been characterized and analyzed in view of the kinetic theory described in the preceding paper. The enzymic condensation reaction between acetyl-CoA and oxaloacetate is subject to substrate-inhibition by acetyl-CoA. This can be attributed to the formation of a productive enzyme-acetyl-CoA complex with a dissociation constant of 110 uM. The binding of acetyl-CoA to the enzyme decreases the on-velocity constant for oxaloacetate-binding from 4000 min-1- micrometer-1 to 1700 min-1-micrometer-1. The affinity of citrate synthase for oxaloacetate increase at least 20-fold on the binding of acetyl-CoA. The latter cooperativity effect can be attributed to a more than 45-fold decrease of the off-velocity constant for oxaloacetate-binding.


The enzymatic synthesis of phosphatidylserine and purification by CM-cellulose column chromatography. Phosphatidylserine has been prepared from phosphatidylcholine by a one-step transphosphatidylation catalyzed by phospholipase D in the presence of L-serine. The resulting mixture of phosphatidylserine and phosphatidic acid is easily and rapidly separated by CM-cellulose column chromatography using step=wise elution with solvents containing increasing percentages of methanol in chloroform. The over-all yield of the procedure is 40-50% depending on the scale of the preparation. CM-Cellulose column chromatography proved to be extremely useful in separating phospholipid mixtures obtained by phosphatidyltransferase reactions of phospholipase D and is also suitable for fractionation of other lipid extracts.


Conformational transitions of antibodies induced by hapten binding. The conformational changes of antibody structure induced by hapten molecule binding were investigated by means of thermal perturbation difference spectroscopy. The studies of the free rabit anti-dinitrophenyl antibodies show the conformational transition at temperatures between 25 and 35 degrees C. The changes occurring at the higher temperature are accompanied by the screening of the significant part of exposed tyrosine residues. Binding of the hapten molecules induces a similar transition to that which occurs between the two temperature dependent states of the free antibody. In contrast to our previous results with anti-dansyl rabbit antibodies the dinitrophenyl lysine stabilizes the "low temperature" native state of the protein. The investigation of the MOPC-315 mouse immunoglobulin A myeloma protein possessing anti-dinitrophenyl activity indicates no conformational transition at temperatures between 25 and 35 degrees C and only a small decrease of tyrosine exposure induced by the hapten binding. Our present and previous results indicate that most of the free immunoglobulins exist in two native conformational states which have a small difference in free energy. Hapten binding causes the transition in equilibrium between the two states towards the one of better binding. It is possible that this transition is necessary but not sufficient step for inducing the effector function of antibodies.


Purification of a bovine counterpart to human prealbumin and its interaction with a bovine retinol-binding protein. A bovine counterpart to human prealbumin was purified from bovine serum by thiol-disulfide exchange chromatography on thiol-Sepharose 4B and affinity chromatography on human retinol-binding protein linked to Sepharose 4B. The bovine prealbumin had alpha1-mobility on agarose gel electrophoresis at pH 8.6. It has the same molecular weight as human prealbumin on gel filtration and consisted of subunits with a molecular weight of 12 500. This is compatible with a tetrameric structure for the bovine protein. Antiserum against human prealbumin cross-reacted with bovine prealbumin and vice versa. The bovine prealbumin formed at high ionic strength complexes with another bovine serum protein which were dissociated at low ionic strength. This property was used to isolate a protein from bovine serum, by chromatography on bovine prealbumin linked to Sepharose which cross-reacted with antiserum against human retinol-binding protein; had a molecular weight of 21 000 and alpha 2-mobility on agarose gel electrophoresis. It was concluded that the latter protein was a bovine retinol-binding protein.


Developmental changes in collagen glycosyltransferase activities in chick embryos. The developmental pattern of collagen galactosyltransferase and collagen glucosyltransferase activities was determined in chick embryos between the 4th and 21st day of growth. Both enzyme activities increased up to the 16th day and decreased thereafter in whole chick embryos and in most tissues studied. The highest collagen glycosyltransferase activities were found in the leg tendons of the 16-day-old embryos, and the activities found in cartilage were higher than those noted in either skin or skull, indicating that the the activities of the collagen glycosyltransferases may play a part in the regulation of the carbohydrate content of the collagen synthesized by a given tissue. The changes observed in the collagen glycosyltransferase activities agree with previous data on the development of prolyl and lysyl hydroxylase activities and also with findings on collagen turnover in the developing chick embryo.


Effect of nitrite upon leghemoglobin and interaction with nitrogen fixation. Nitrite (0.4 mM) added to soybean bacteroid preparations strongly inhibited C2H2 reduction. In the presence of leghemoglobin (0.1mM), a 3-fold enhancement of nitrogen fixation occurred but the inhibitory effect of nitrite was delayed. Spectra of leghemoglobin showed a rapid disappearance of the 574 nm and 541 nm peaks of oxyleghemoglobin the presence of nitrite. Concomitant oxidation of this hemoprotein gave ferric leghemoglobin as the single final product. High nitrite levels could depress nitrogen fixation both by inactivation of nitrogenase and by conversion of leghemoglobin into an inactive form. Nitrite present at low concentrations reacts with this hemoprotein and is then no longer able to penetrate into bacteroids.


Bioaffinity chromatography of thyrotropin using immobilized concanavalin A. The behavior of crude preparations of whale and bovine thyrotropins was studied on an affinity column packed with concanavalin A-Sepharose. Only a small portion of the proteins applied was adsorbed to the column and eluted quantitatively with 0.5 M methyl-alpha-D-glucoside or -mannoside. Immunoreactive as well as hormonally active thyrotropin was recovered exclusively in the absorbed fraction. The usefulness of this chromatographic procedure for the group separation of pituitary glycoprotein hormones is discussed.


Alteration of human placental lactogen by mammary gland. Human placental lactogen was prepared in high purity and in good yield applying a minimum of purification steps. The isolated hormone was characterized with respect to isoelectric and electrophoretic properties, molecular size (estimated mol. wt. 23 000) and stability to heat, pH and organic solvents. Investigation of in vitro interaction between placental lactogen and mammary gland (mouse, rat) revealed a rapid alteration of hormone with loss of immunoreactivity resulting. The target organ as a selective alteration site of placental lactogen was suggested by a lack of similar action on the hormone by a number of other tissues tested, including liver, kidney and lung. The reaction involving hormone alteration by mammary gland was localized to a particulate-bound enzyme, sedimentable at 10 000 X g and undissociated by sonication in 0.5% Triton X-100. Examination of the reaction products revealed hormone degradation with formation of diffusible components and loss of original electrophoretic identity as well as immunoreactive properties. The reaction characteristics included: pH optimum between 7.5 and 8.0, an absolute salt requirement (NaCl, KCl, at concentration greater than 0.15 M for maximal activation), inhibition by Cleland's reagent and lack of reaction interference by pituitary prolactin.


Purification and specificity of antibodies to inosine 5'-monophosphate. Antibodies to inosine 5'-monophosphate elicited in rabbits by immunization with a conjugate of IMP (oxidized with periodate) and bovine serum albumin have been purified by affinity chromatography. By the use of two affinity columns, Sepharose-IMP and Sepharose-oligo(I), the antibodies have been fractionated into three fractions. By gel diffusion, the three fractions were found to react with the conjugates of bovine serum albumin and IMP, GMP and AMP respectively. The association constants for the binding of the Fab fragments purified on the Sepharose-oligo(I) column and several haptens have been deduced from fluorescence experiments. It is shown that the base and the phosphate group play an important part in the binding of IMP to Fab fragments. No reaction has been found between the antibodies and poly(I).poly(C) by gel diffusion. However, the antibodies interact with poly(I).poly(C) since they decrease the thermal stability of poly(I).poly(C).


Isolation of nematode inhibitor from hemolymph of the snail, Helix aspersa. Hemolymph plasma of the snail Helix aspersa which inhibits maturation and reproduction of its mantle cavity-inhabiting nematode, Rhabditis maupasi, was separated biochemically for the active proteinaceous component. Isolation of the active inhibitor was performed using ion-exchange chromatography in combination with subsequent gel filtration. The isolated peaks were bio-assayed in vitro on nematode larvae. The fractions harboring inhibitory protein suppressed larval growth and adult reproduction in vitro. The isolated fraction was purified by gel filtration and characterized on the basis of a single band on starch zone electrophoresis and positive reaction only with folin-phenol reagent.


[Comparative study of histones from slime mold Physarum polycephalum and calf thymus]. The histones from slime mold Physarum polycephalum and calf thymus were characterized in terms of some physico-chemical properties. The molecular weights of six principal histone fractions of Ph. polycephalum were found to be the following: P1--22 700, P3--15 700, P4a--15 000, P4b--14 300, P5--12 800 and P6--10 500. Electrophoretically homogenous histone fractions H1, H2b and H4 of calf thymus and histones P1, P3, P4b and P6 of slime mold were obtained by gel-filtration on Acrylex P-60. These findings suggest that fractions P1, P4a, P4b, P5 and P6 of slime mold Ph. polycephalum are homologus with respect to the histone fractions H1, H3, H2b, H2a and H4 of calf thymus. Only fraction P3 has no corresponding fraction in the calf thymus histones; a fraction corresponding to histone P3 of slime mold was absent.


Agglutinins and lysins in the molluscan family Planorbidae: a survey of hemolymph, egg-masses, and albumen-gland extracts. 1. Thirty-one strains representing eight species and four genera of the molluscan family Planorbidae were surveyed for the presence of hemagglutinins and hemolysins. Extracts prepared from albumen-glands and egg-masses, as well as hemolymph, were assayed by a micro-hemagglutination technique in which human erythrocytes were used as receptors. 2. Hemagglutinins and hemolysins for human erythrocytes were not uniformly present in all members of the family and detection of these substances depended, in part, on the material tested. 3. Neither heating to 56 degrees C, dialysis, storage at -20 degrees C for up to a year, nor repeated freezing and thawing appeared to effect the agglutinating activity of egg or albumen-gland extracts. 4. Inhibition of the agglutinating activity of Helisoma hemolymph could be accomplished with N-Acetyl-D-Galactosamine, but this sugar had no effect on the activity of Biomphalaria agglutinins. 5. There is evidence to suggest that the source and nature of agglutinins from Helisoma and Biomphalaria species are different. 6. Lectins may be of some value in characterizing snail populations and as an aid in the taxonomic discrimination of species.


[Homology of histones H2B from calf thymus and P4B from slime mold Physarum polycephalum]. A comparative study of the amino acid composition of histone fractions P4b from slime mold Physarum polycephalum and H2B from calf thymus was carried out using peptide mapping. It was shown that 75% of peptides are common for both proteins. The slime mold histones contain two fractions (P4B and P3), which are homologous to the H2B histone fraction of calf thymus. The data of amino acid analysis, peptide mapping and some physico-chemical properties of the histones revealed the following correlation of the two types of histone fractions: P1--H1, P4a--H3, P4b and P3--H2B, P5-H2A, P6--H4.


[A method for determination of alkaline sucrose gradient sedimentation patterns of DNA for nondividing and slowly dividing cells]. A radiometric method for developing the alkaline sucrose gradient sedimentation patterns of DNA from non-labeled cells is described. The method is based on labeling of DNA in the gradient fractions by means of a labeled amino acid binding to DNA in the presence of formaldehyde. The procedure involves incubation of the fractions with a mixture of the labeled amino acid and formaldehyde, filtration of the incubation mixtures through nitrocellulose filters and radiometry of the filters. Relationship between the radioactivity on the filter and DNA concentration in the sample is linear. DNA detection sensitivity and specificity are satisfactory for the use under gradient ultracentrifugation condition. The non-labeled cells DNA sedimentation patterns developed by the method described and those of DNA from the same cells labeled with 3H-thimidine in vivo are found to be similar.


Intensification of the alcohol withdrawal syndrome following periodic electroconvulsive shocks. Repeated electroconvulsive shocks (ECSs) administered once every 3 days to rats at either of two current intensities led to a progressive intensification of the motor seizure pattern. Moreover, the incidence of convulsive symptoms elicited by subsequent alcohol exposure and withdrawal was greatly increased by prior exposure to the repeated ECSs. These results illustrate a treatment--drug interaction which could have hazardous consequences for patients undergoing electroconvulsive therapy. Thus, until the appropriate tests can be conducted on human patients, drug intake following electroconvulsive therapy should be carefully regulated.


Maternal alcohol and neonatal health.

Maternal alcohol and neonatal health. The disappearance of alcohol from blood following a single dose is a function of age, but by the time the animals are 90 days old the results are consistent from animal to animal. A single dose given to a pregnant female is distributed between fetus and maternal tissues and fluids, with a portion of the alcohol trapped in the amniotic fluid. Kittens born to mothers receiving alcohol during the last 2 weeks of pregnancy demonstrate hyperactivity, small size with unimpaired growth rate, and slow maturation of the righting reflex. It would require more experimentation to decide whether these changes are due to alcohol withdrawal or to toxic effects of the alcohol per se.


Lithium inhibition of norepinephrine and dopamine receptors. Sprague-Dawley rats (150-200 g), serving as their own controls, were evaluated for hyperactivity (HyA) and stereotyped behavior (SB) produced by chronic (1 week) ip administration of different doses of drugs that have differential NE and DA " agonist" properties. Drugs utilized were d- and l-amphetamine, metamphetamine, cocaine, and the specific and direct DA receptor agonist apomorphine. This was followed by 1 week of rest after which all rats received 3 weeks of lithium chloride (Li) subcutaneously. In the last week of Li administration, each rat also was given the same drug administered previously at the same dose. In all cases, Li decreased both HyA and SB, suggesting Li effects both at the pre- and postsynaptic neuron (receptor).


Histochemical study of cholinergic activities in exocrine pancreas of dogs. Modifications related to chronic alcoholism. An histochemical study has been made of pancreatic biopsies from 5 dogs with gastric and pancreatic Thomas cannulae. 3 dogs received 2 g kg-1 day-1 ethanol for 3, 4 and 48 months; 2 dogs which did not received alcohol served as controls. Acetylcholinesterase has been studied by the histochemical method of Koelle. Choline acetyl transferase by the method of Burt. Our results suggest that there is less acetylcholinesterase activity in the pancreas of alcoholic dogs than in control dogs. This is as more pronounced when the alcohol consumption is prolonged. On the contrary, the intensity of the histochemical reaction for choline acetyl transferase is greater in alcoholic dogs than in controls. Although histochemical methods are only semi-quantitative, these results suggest that the cholinergic tone of the exocrine pancreas is increased in chronic alcoholic dogs. This has already been suggested by previous physiological experiments performed on the same animals.


Spectral studies on the conformation of rhodopsin. A summary is given of visible and ultraviolet spectral studies which deal with the conformation of rhodopsin in situ and solubilized with detergent. Emphasis is placed on studies which give specific information about the macromolecular structure and which set quantitative limits on the magnitude of light-induced conformational changes.


Mass spectrometry of Alternaria mali toxins and related cyclodepsipeptides. The structures of AM-toxins I, II and III, host specific phytotoxic metabolites of Alternaria mali, can be readily deduced from low and high resolution mass spectral data, since the amino acids and their sequences are demonstrated by this technique. Additionally, the general fragmentation of these compounds by electron impact is discussed by comparing the spectra of analogous synthetic compounds.


[Antagonism between L-DOPA and apomorphine in their effects on rat behavior]. The effect of simultaneous injections of apomorphine and l-DOPA on stereotypy, aggressiveness, on the exploratory motor activity, the threshold of emotional reactivity and aggressiveness elicited by painful electrical stimulation was studied in experiments on male albino rats. When injected separately, in control experiments, both compounds had similar effects on the exploratory-motor activity and the emotional behaviour, but when injected simultaneously in various doses a distinct antagonism between l-DOPA and apomorphine, according to all the behaviour tests, was noted (a decrease of sterotypy, aggressiveness and emotional reactivity). Against the background of l-DOPA apomorphine (5 mg/kg) increased the dopamine content in the forebrain and in the diencephalon even more. It is suggested that the increased level of the functionally active mediator suppressed the activity of postsynaptic receptors sensitive to it.


[Low-molecular homogeneous thymus fraction stimulating immunogenesis]. The influence of purified acetic acid thymus extract (thymarin) and its three fractions isolated by ion-exchange chromatography upon immune response to SRBC in mice was studied. Each fraction was identified as homogeneous individual material of polypeptide nature. Crude thymus extract and one of its fractions with an approximate molecular weight of 5000 exerted a strong stimulating effect on the thymus-dependent response: formation of IgM- and IgG-antibody producing cells and on the circulating antibody level.


[Antigen-dependent induction of a non-specific humoral factor blocking rosette-forming cells in experiments in vitro and in vivo]. Isologous serum of CBA mice immunized with rabbit immunoglobulins (ARIS) contained a factor capable of inactivating rosette-forming splenocytes (RFC) in vitro from the same strain of mice immunized with SRBC. When mouse SRBC immunization was carried out against the background of ARIS injection the court of RFC to SRBC at the peak of immune response was 30% of that of mice injected with SRBC and normal isologous serum. A decrease of RFC count was the result of disappearance of the theta-negative RFC. Passive ARIS immunization failed to influence the antigen-induced proliferation of the antibody-forming cells and the synthesis of antibodies against SRBC.


C-fragment of lipotropin--an endogenous potent analgesic peptide. 1 A series of peptides derived from porcine lipotropin was examined for analgesic and other morphine-like properties on infusion into the cannulated third ventricle of cats.2 Lipotropin (LPH 1-91) itself produced no analgesia or other morphine-like effects when infused in a dose of 150 mug.3 C-fragment (LPH 61-91) produced strong long-lasting analgesia when infused in a dose of 10 or 20 mug; on a molar basis the potency was between 90 and 180 times that of morphine. The following morphine-like effects were also produced: shivering leading to fever, vasodilatation of the pinnae, mydriasis, opening of the palpebral fissures, tachypnoea with bouts of panting, vocalization, hyperexcitability, restlessness and catalepsy. All the effects, including analgesia, were abolished by an intraperitoneal injection of naloxone (1 mg/kg).4 Hyperglycaemia, another central effect produced by morphine, was obtained with C-fragment infused in a dose of 60 mug.5 On intravenous injection, C-fragment produced analgesia with a dose of about 200 mug/kg. Administered by this route, C-fragment was again more potent than morphine.6 C'-fragment (LPH 61-87), LPH 61-78 and LPH 61-69, either had no analgesic effect or produced weak short-lasting analgesia when infused in doses up to 100 mug.7 Methionine enkephalin (LPH 61-65) either produced very weak short-lasting analgesia or had no analgesic effect when infused in doses of between 30 and 400 mug.8N-methyl methionine enkephalin amide in which both termini of methionine enkephalin were protected against degradation by exopeptidases produced long-lasting analgesia when infused in doses of 150 to 180 mug; its analgesic potency was approximately 100 times less than that of C-fragment. Blocking only one terminus of methionine enkephalin did not appear to endow the peptide with analgesic properties. The N-methyl pentapeptide amide produced other morphine-like effects of which the most striking was catalepsy. All the effects were abolished by intraperitoneal naloxone (1 mg/kg).


Electrochemically stimulated release of luteinizing hormone and ovulation after surgical interruption of lateral hypothalamic connections in the rat. Plasma LH and FSH were studied in adult female rats following bilateral electrochemical stimulation (ECS) of the medial preoptic area (MPOA). By stereotaxic surgery frontal (FC) and frontal-lateral (LFC) retrochiasmatic "deafferenting" cuts were made with a Halász knife (1.5 mm radius). At 3 and 10 weeks after surgery rats were given pentobarbital (32 mg/kg, i.p.) at 13:30h and stimulated at 15:00h with anodal direct current (20 muA for 60 sec) via concentric bipolar steel electrodes placed bilaterally 0.9 mm from the midline. Stimulation at 3 weeks after FC increased plasma LH from a prestimulation level of 95 +/- ng/ml to 227 +/- 51 ng/ml 80 min after ECS, from which it fell to 111 +/- 29 ng/ml at 160 min, with 2 of 7 animals ovulating the next day. LFC females had similar pre-ECS plasma LH levels 3 weeks after surgery (71 +/- 10 ng/ml), but LH concentrations at 80 min (104 +/- 21 ng/ml) and 160 min post-ECS were significantly lower than those of FC rats, and 0 of 5 rats ovulated. Following a similar protocol 10 weeks after surgery, stimulating the MPOA resulted in comparable elevations in plasma LH and 4 of 10 FC animals ovulated; however, the LFC group still retained a significant blocking effect on ovulation (only 1 of 12 ovulated) when compared with controls the next day. Plasma FSH concentrations were not significantly altered by MPOA stimulation at the parameters employed, either before or after deafferentation. However, LFC resulted in reduced ovarian and uterine weights when compared with controls at both 3 and 10 weeks, whereas FC exerted no observable effect on these organs at these intervals of study. The results of these studies suggest that lateral input to the media basal hypothalamus contributes to MPOA mediated release of LH and ovulation as well as to tonic maintenance of ovarian and uterine function.


Urease activity in the digestive tract of the chick and its role in the utilisation of urea as a source of non-amino nitrogen. Urease activity was found in caecal contents (about 10 mg urea metabolised/g h) and crop contents (about 0-5 mg urea metabolised/g h):there was very low activity in the contents of the colon but none in the rest of the digestive tract. 2. The urease activity of the crop contents was not bacterial in origin but the soyabean meal contained in the diet was found to have comparable activity. 3. Diets low in non-essential nitrogen and based on soyabean, fish meal or fish meal plus 0-2% jack bean urease, did not support higher growth rates when supplemented with urea. 4. The livers of chicks fed on the diet containing fish meal, urea and urease had significantly higher concentrations of free non-essential amino acids than those of chicks fed on the same diet but with urease excluded This suggested that dietary urease affects the availability of ammonia for the synthesis of non-essential amino acids but not for growth.


Use of 13C in biosynthetic studies. The labelling pattern in tenellin enriched from isotope-labelled acetate, methionine, and phenylalanine. The biogenetic origin of the carbon atoms in tenellin has been established by adding 13C-enriched compounds to cultures of Beauveria bassiana, and determining the isotopic distribution in the metabolite by 13C nuclear magnetic resonance spectrometry. Tenellin is formed by condensation of an acetate-derived polyketide chain with a phenylpropanoid unit that may be phenylalanine. Alternate carbon atoms of the polyketide chain were labelled with sodium [1(-13C)]- and [2-(13C]-acetate; sodium [1,2-(13C)]acetate was incorporated as intact two-carbon units, the presence of which in tenellin was apparent from coupling between adjacent 13C-enriched carbons. Substituent methyl groups of the polyketide-derived alkenyl chain were labelled with L-[Me-13C]methionine. The labelling patterns from DL-[carboxy-13C]phenylalanine and DL-[alpha-13C]phenylalanine indicated a rearrangement of the propanoid component at some stage in the synthesis. The mass spectrum of tenellin from cultures administered L-[15N]phenylalanine showed isotopic enrichment similar to that obtained with 13C- or 14C-labelled phenylalanine. During incorporation of L-[carboxy-14C, beta-3H]phenylalanine 96% of the tritium label was lost, discounting the possibility of a 1,2-hydride shift during biosynthesis of the metabolite.


Biosynthesis of gibberellins in Gibberella fujikuroi. Gibberellin A47. Two minor radioactive products in cultures of G. fujikuroi strain ACC917 supplemented with labeled gibberellin A4 have been identified as the 2alpha-hydroxy derivative, gibberellin A47, and the hydrate, gibberellin A2. In addition, evidence was obtained by combined gas chromatography--mass spectrometry for the formation of 3-O-acetylgibberellin A1,3-O-acetylgibberellin A3, and gibberellin A20 by the culture. The time course of gibberellin synthesis in defined and complex media, as well as changes in the relative amounts of radioactive gibberellins formed when [4C]gibberellin A4 was added to cultures at different ages, suggest that the composition of the gibberellin mixture produced is determined by the balance between synthesis and decay of key enzymes in the branching pathway.


Role of orthophosphate concentration in the regulation of ribose phosphate synthesis and purine metabolism in Ehrlich ascites tumor cells. Concentrations of intracellular orthophosphate were determined in Ehrlich ascites tumor cells incubated with glucose, inosine, or uridine in media of different orthophosphate concentration. The effects of orthophosphate concentration on the accumulation of lactate and of phosphoribosyl pyrophosphate and on concentrations of ribose 1-phosphate and ribose 5-phosphate in tumor cells incubated with glucose were also determined. Both the phosphorolysis of inosine and the rate of catabolism of ATP in cells incubated with 2-deoxyglucose were also influenced by the orthophosphate concentration of the medium.


The relationship between rhythmic hyphal growth and circadian formation of sclerotia in Sclerotium rolfsii Sacc. The mycelium of the phytopathogenic fungus Sclerotium rolfsii type R (ATCC 26326) grown on a solid medium supplemented with 70 mM l-threonine exhibits a synchronous formation of sclerotia in circles whereas both leading and lateral hyphae enter into a rhythmic growth rate. A rhythmic activity of malate synthetase (malate synthase, EC 4.1.3.2) which accompanied the hyphal rhythm and preceded the formation of sclerotial circles was demonstrated. Both translocation of L-[14C]threonine and incorporation of L-[14C]leucine were significantly related to the circadian formation of sclerotia.


Effect of rj1rj1 (non-nodulating) soybeans on nodulation of near isogenic Rj1Rj1 plants in nutrient culture. An earlier proposal (Can. J. Microbiol. 7: 851; 1961) that rj1rj1 (non-nodulating) soybeans (Glycine max (L.) Merr.) excrete a substance that inhibits nodulation of Rj1 Rj1 (nodulating) plants was tested. Using near isogenic lines (isolines) of "Clark" and "Harosoy" soybeans, we consistently found nonsignificant reduction in nodule number and acetylene reduction per Rj1Rj1 plant grown in association with their rj1rj1 counterparts: these results suggest that a nodulation inhibitor is not associated with the rj1 gene. Reducing the number of plants grown in each pot produced significant (P = 0.05) reductions in nodule number per Rj1Rj1 plant, and resembled the observations of the earlier report. On this basis, we suggest that the reported inhibition of nodulation was due to a failure to detoxify or remove an inhibitor (possibly nitrate) already present in the nutrient solution. Both Clark isolines removed nitrate from their nutrient solutions at similar rates. Harosoy rj1rj1 plants removed nitrate at a significantly (P - 0.05) slower rate than Harosoy Rj1Rj1 plants, but the differences were not correlated (P = 0.05) with the small observed decreases in nodulation. These differences in nitrate uptake were highly correlated (P = 0.01) with reduced dry weight per Harosoy rj1rj1 plant.


Growing human sarcomas in culture.

Growing human sarcomas in culture. Efforts to establish human soft tissue and bone sarcomas in long-term cultures are described. Of 49 attempts 22 (45%) were successful, yielding cells that proliferated in vitro for 6 months or longer. Infrequent medium changes, maintenance of pH in physiologic range and avoidance of early trypsinization all appear to be important for survival of explanted cells and their initial growth under culture conditions. Cytologic criteria can be used to confirm that cells in established lines derive from malignant mesenchymal cells present in the tumors of origin.


Population kinetics of an Ehrlich ascites tumor following treatment with methylglyoxal-bis(guanylhydrazone), a polyamine synthesis inhibitor. Population kinetics of Ehrlich ascites tumor cells grown in vivo were studied following treatment with methylglyoxal-bis(guanylhydrazone) (MeGAG; NSC-32946), a potent chemotherapeutic agent and a specific polyamine synthesis inhibitor. MeGAG-treatment resulted in a continuous accumulation of cells in the S and G2 phases of the cell cycle, and, as a consequence, the rate of cell proliferation decreased. This finding emphasizes the importance of the polyamines for progression through the cell cycle.


Potentiation of the action of azaserine on the rat pancreas by raw soya bean flour. Rats were treated with raw soya flour to produce pancreatic hypertrophy and also received azaserine, a pancreatic carcinogen. The combined treatment resulted in the development of large numbers of hyperplastic nodules in the acinar tissue of the rat pancreas. We conclude that the pancreas of the rat stimulated to proliferate by raw soya flour provides a sensitive model for detecting and studying pancreatic carcinogens.


RNA metabolism of murine leukemia virus: size analysis of nuclear pulse-labeled virus-specific RNA. A system for excess DNA hybridization of Moloney murine leukemia virus (M-MuLV)-specific RNA from infected mouse cells with M-MuLV cDNA immobilized on nitrocellulose filters was developed. In the presence of unlabeled heterologous rabbit liver RNA, 0.3-0.5% of labeled, infected cell nuclear RNA bound to the filters, while 0.05% or less of nuclear RNA from uninfected cells bound. Sedimentation analysis of pulse-labeled nuclear RNA was performed, and hybridization across sucrose gradients indicated that the major pulse-labeled, virus-specific RNA was 38S, similar or identical in sedimentation to the virion subunit RNA. A minor component of pulse-labeled, virus-specific RNA larger than 38S, was detected (40-60S), but kinetic experiments indicated that it was not an obligate precursor to 38S virus-specific RNA. Simultaneous analysis of steady state and pulse-labeled, virus-specific nuclear RNA across sucrose gradients indicated that the 38S virus-specific RNA was not detectably different from the steady state "35S" nuclear RNA previously identified. More detailed resolution on agarose gels also showed no difference. Thus the primary transcript of M-MuLV-specific RNA appears to be 38S, the same size as stable cellular virus-specific RNA, and no evidence for a higher molecular weight precursor was found.


Heterogeneity of the ribosomal genes in mice and men. The structures of mouse and human ribosomal DNA were studied using the restriction enzymes Eco R1 and Hind III. Individual mice or humans showed a heterogeneous pattern of restriction fragments resulting from differences in the non-transcribed spacer DNA. Six individual mice from the inbred strain CBA/H-T6 had identical patterns. The same pattern was shown by another CBA strain and by C3H. These strains were originally derived from a BALB X DBA cross made in 1920. Different patterns were found for BALB/c, C57BL and Mus poschiavinus. Cultured cells derived from C3H mice (L cells) showed a pattern quantitatively different from that of the parent strain, but two myeloma cell lines derived from BALB/c showed the same pattern as BALB/c mice. Ribosomal DNA in man is also heterogeneous. Differences were observed between human DNAs in the amounts of the different spacer classes. Studies on mouse-human cell hybrids suggest that some spacer classes are present on more than one of the five human nucleolus organizers.


Mouse L cell mitochondrial DNA molecules are selected randomly for replication throughout the cell cycle. The number of mitochondrial DNA molecules in a cell population doubles at the same rate as the cell generation time. This could occur by a random selection of molecules for replication or by a process that ensures the replication of each individual molecule in the cell. We have investigated the rate at which mouse L cell mitochondrial DNA molecules labeled with 3H-thymidine during one round of replication are reselected for a second round of replication. Mouse L cells were labeled with 3H-thymidine for 2 hr, chased for various periods of time and then labeled with 5-bromodeoxyuridine for 4 hr immediately before mitochondrial DNA isolation. A constant fraction of 3H-thymidine-labeled mitochondrial DNA incorporated 5-bromodeoxyuridine after chase intervals ranging from 1.5-22 hr. This result demonstrates that mitochondrial DNA molecules replicated in a short time interval are randomly selected for later rounds of replication, and that replication of mitochondrial DNA continues throughout the cell cycle in mouse L cells.


A very large repeating unit of mouse DNA containing the 18S, 28S and 5.8S rRNA genes. The organization of the 18S, 28S and 5.8S rRNA genes in the mouse has been elucidated by mapping with restriction endonucleases Eco RI, Hind III and Bam HI. Ribosomal DNA fragments were detected in electrophoretically fractionated digests of total nuclear DNA by in situ hybridization with radioiodinated rRNAs or with complementary RNA synthesized directly on rRNA templates. A map of the rDNA which includes 13 restriction sites was constructed from the sizes of rDNA fragments and their labeling by different probes The map indicates that the rRNA genes lie within remarkably large units of reiterated DNA, at least 44,000 base pairs long. At least two, and possibly four, classes of repeating unit can be distinguished, the heterogeneity probably residing in the very large nontranscribed spacer region. The 5.8S rRNA gene lies in the transcribed region between the 18S and 28S genes.


'Cleavage fields': hypothesis on early embryonic development. The hypothesis is put forward that events of the early embryonic development can be interpreted on the basis of a radial distribution of cytoplasmic components, i.e. of a 'cleavage field', progressively established during the growth of the oocyte. The orientations of the cleavage spindles and the corresponding furrows' positions are assumed to be correlated to the field's temporal evolution which, in turn, is determined by flows of cytoplasmic components originated by the changes in the membrane shape. From this viewpoint, a simple explanation of egg regulation is proposed, and the particular case of the sea urchin embryo is briefly discussed.


Differences in agglutinability of adult and fetal human fibroblasts using phytohemagglutinin. Whereas Concanavalin A (Con A) and Wheat Germ Agglutinin (WGA) detect differences in the agglutinability of transformed, established and secondary cultures, Phytohemagglutinin (PHA) detects differences between cultured adult and fetal human fibroblasts. Adult cells agglutinate with PHA to the same extent as transformed cells, whereas fetal cells show significant agglutination only after trypsinization. Differences in cell size, growth rate, surface architecture or binding of fluorescent PHA could not be demonstrated between adult and fetal cells. Although the basis for this apparent difference in agglutinability remains unknown, it is the first demonstration that fetal cells (even after prolonged in vitro culture) retain at least some surface properties not shared by adult or transformed cells.


Effects of mercuric chloride on synchronized Chinese hamster ovary cells: survival and DNA replication. Chinese hamster ovary (CHO) cells in vitro were treated with HgCl2 at various stages in the cell cycle and the effects of this chemical on cell survival, DNA replication, and cell division were observed. In terms of survival the early G1 cells were the most sensitive to treatment, followed by late G1 and early S, while mid S and late S-G2 treated cells were the least sensitive. Treatment with HgCl2 also resulted in reduced rates of DNA replication and delays in cell division. The early G1 treated cells showed substantially reduced rates of DNA replication followed by 4--5 h division delay. The early S and late S-G2 treated cells had some reduction in their rates of DNA replication followed by corresponding division delay of 2.5 h in the early S treated cells and 1 h in the late S-G2 treated cells.


Quantitative method for estimating myoglobin in urine. A simple, rapid, and quantitative immunochemical technique is described for quantitation of urinary myoglobin. The test is specific for myoglobin in the presence of hemoglobin or tissue constituents, can be completed within 1 h or less, and can detect 1 to 2 mg/liter. Normal urinary myoglobin concentrations are less than this.


Genetic variance of erythrocyte parameters in adult male twins. Erythrocyte count, hematocrit, mean corpuscular volume and reticulocyte counts were measured in 59 pairs of monozygotic (MZ) and 69 pairs of dizygotic (DZ) adult, Caucasian male twins. The means of MZ and DZ twins were not significantly different for any of the traits measured. Erythrocyte count and mean corpuscular volume had significant estimates of genetic variance (P less than 0.05).


Humoral response in blood group P phenotype persons subjected to an avian environment. The avian environment contains among many antigens, the blood group substance P1-P1 antigen is present in certain Enterobacteriaceae found in bird droppings. Examples include Enterobacter, Klebsiella, E. coli, Enterobacter Haffniae and Proteus. P2 persons exposed to the avian environment may develop anti-P1 activity. A greater percentage of precipitins to bird antigens was found in P2 than in P1 persons. In P1 persons seven out of twenty-eight (25%) had precipitins to bird droppings. In P2 persons, ten out of thirteen (77%) had anti-P1 activity and nine out of thirteen (69%) had precipitins to bird droppings. This greater response in P2 persons may be explained by the constant stimulation of these persons lacking the P1 antigen that triggers more response with other antigens in addition to the P1 antige. In the study of immunological responses to inhaled antigens, the blood group of the patients should be determined in order to assess the importance of constant stimulation by foreign material in persons lacking some blood group antigens.


Metabolism of pentaerythritol trinitrate. The absorption, excretion, and biotransformation of 14C-labeled pentaerythritol (PE) trinitrate was studied in man. The administration of a single sublingual dose was followed by rapid absorption and extensive biotransformation. Six drug metabolites were identified. Final excretion of the drug and its metabolites was almost totally through the kidney. Low levels of unchanged drug were present in plasma and urine. PE mononitrate was the major drug metabolite in plasma and urine. Glucuronides of PE trinitrate, dinitrate, and mononitrate were identified for the first time in man. PE trinitrate glucuronide appeared in plasma rapidly and about 8% of the dose was excreted in urine. Reversible and irreversible pathways are proposed for the formation of the metabolites. The reconversion of PE trinitrate glucuronide to PE trinitrate is postulated to explain the duration of drug activity and excretion.


Lipid hemodialysis versus charcoal hemoperfusion in imipramine poisoning. Previous experimental results have demonstrated the possibility of eliminating imipramine (14 C-IP) by hemodialysis. A simultaneous uptake of the substance by the polyvinyl chloride extracorporeal blood lines could be shown. Based on these results the imipramine absorption capacity of the blood lines and of the artificial kidney (Hollow Fiber Artificial Kidney, HFAK, Model 4) were studied. Imipramine (IP) absorption capacity of a usual blood-line set (arterial and venous, surface area 86,000 mm) was estimated to be 43 mg, and that of the HFAK to be 207 mg. Charcoal hemoperfusion (300 gm of coated activated coconut charcoal) eliminated more than 90% of IP from the blood within 3 hr if the initial IP blood concentration was 2 mg/ml. In comparison, lipid hemodialysis using 20% soybean oil as dialysate eliminated 95% of IP from the blood when the initial IP blood concentration was 1 mg/ml and 98% when the initial IP blood concentration was 2 mg/ml. In vivo studies on the elimination of IP by lipid (10%) hemodialysis demonstrated a substantial removal of the substance. Within 2 hr of treatment, 12% of the administered dose (75% of the LD50) was eliminated.


Design and synthesis of parathyroid hormone analogues of enhanced biological activity. Three analogues of bovine parathyroid hormone (bPTH), [Tyr-34]bPTH-(1--34) amide, [Nle-8,Nle-18,Tyr-34]bPTH-(1--34)amide, and [Nle-8,Nle-18, o-NPS-Trp-23,Tyr-34]bPTH-(1--34)amide were synthesized by the solid-phase method. The synthetic peptides were found to be homogeneous in multiple analytical systems. These analogues represent the NH2-terminal one-third of the hormone, previously shown to contain all the structural requirements necessary for biological activity, but containing several structural modifications associated with enhancement and stabilization of biological activity. When tested in the in vitro renal adenylylcyclase assay, in which unsubstituted bPTH-(1--34) has a potency of 5400 MRC Units/mg, [Tyr-34]bPTH-(1--34)amide had a potency of 16,1000 MRC Units/mg, [Nle-8,Nle-18,Tyr-34]bPTH-(1--34)amide was 10,100 MRC Units/mg, and [Nle-8,Nle-18,o-NPS-Trp-23,Tyr-34]bPTH-(1--34)amide was 10,600 MRC Units/mg. The diverse structural features incorporated in these hormone analogues, resulting in a several-fold increase in biological activity in vitro, demonstrate additive effects on biological activity and should prove valuable in certain biological applications, as well as in the design of other parathyroid hormone analogues of enhanced potency.


A new concept for control of the estrous cycle of the ewe based on the temporal relationships between luteinizing hormone, estradiol and progesterone in peripheral serum and evidence that progesterone inhibits tonic LH secretion. The temporal relationships between LH, estradiol and progesterone in peripheral serum of the ewe were characterized throughout the estrous cycle. Between successive preovulatory LH sruges, serum concentrations of LH fluctuated markedly in a manner indicative of pulsatile discharges. Mean serum LH and progesterone concentrations were inversely related, LH being highest during the early and late luteal phases of the estrous cycle and lowest in the mid-luteal phase. A progressive, 5-fold increase in serum LH concentrations occurred between the onset of the precipitous fall in circulating progesterone attendant to luteolysis and the initiation of the preovulatory LH surge. Two major increments in circulating estradiol were observed in each cycle, both occurring when serum LH concentrations were relatively high. One estradiol increment occurred during the early luteal phase, the other during the 2-3 days prior to onset of the preovulatory LH surge. The latter estradiol increment thus accompanied the progressive, 5-fold increase in circulating LH which precedes the LH surge. The observations are inconsistent with the view that tonic LH secretion in the ewe is solely a function of a negative feedback action of estradiol. Rather, the temporal relationships between circulating hormones, in conjunction with recent findings that progesterone can inhibit tonic LH secretion in the ewe, lead to the conclusion that progesterone plays a major role in the regulation of tonic LH secretion during the estrous cycle of sheep.


Joint-ill and other bacterial infections on thoroughbred studs. From a perinatal survey carried out on Thoroughbred studs, information was obtained on the incidence, mortality and long-term effects of joint-ill and other bacterial infections. Many of these infections occurred in foals with a history of other perinatal abnormalities, and in foals in which receipt of colostrum was delayed. There was no direct relationship between the incidence of infection and maternal age, but prenatal lactation and loss of colostrum were more frequent in older mares. There was a significant reduction in the incidence of infections in foals given neomycin and framomycin during the first 30 days of life.


Immobilised lipoamide dehydrogenase. 2. Properties of the enzyme immobilised to agarose through spacer molecules of various lengths. 1. Pig heart lipoamide dehydrogenase (NADH: lipoamide oxidoreductase, EC 1.6.4.3) has been immobilised to Sepharose by thiol-disulphide interchange via a series of thiolated spacer molecules of increasing length. A number of properties of the immobilised enzyme have been investigated in order to ascertain the effects of proximity to the matrix backbone. 2. Proximity to the matrix backbone reduced the specific activity for lipoamide as substrate but enhanced by 3-8-fold the diaphorase activity with 2,6-dichloroindophenol. These observations are explained in part by an increase in the apparent Km for lipoamide when the enzyme is covalently attached to Sepharose via a short spacer molecule. 3. Both the thermal stability at 90 degrees C and the stability in 30% (v/v) dioxane are enhanced by up to 200% when the enzyme resides close to the matrix but approach those of the native enzyme as the length of the spacer molecule is increased. 4. These data have been correlated with measures of the accessibility of the enzyme as the nominal length of the spacer arm was increased. Thus, as the chain length increased, the rate of cleavage of the disulphide linkage between the enzyme and spacer increased and the enzyme became more susceptible to proteolysis by thermolysin. In contrast, increasing the chain length of the spacer made the enzyme less amenable to inhibition by a specific antibody. 5. These data are discussed in terms of the effect of the matrix on the conformation of the bound enzyme.


Coprophagia as seen in thoroughbred foals. Four Thoroughbred foals were seen to quickly eat part of the faeces deposited by their own dams on some 40 per cent of the mare-defaecating occasions observed between the second and fifth week after birth. They did not do it before or after this period. This behaviour was thought to be a feeding pattern which formed a normal part of the foal's development.


The efficacy of fenbendazole in the control of immature strongyle infections in ponies. The efficacy of fenbendazole against immature stages of Trichonema spp., Strongylus vulgaris and Strongylus edentatus was evaluated. Naturally infected 6 to 12 month old ponies were given single, oral doses of 0, 15, 30 and 60 mg/kg of body weight. A dose response relationship was noted between increasing dose levels and efficiency against larval trichonemes and migrating stages of S. vulgaris and S. edentatus. Dose levels of 30 mg/kg and higher removed 93 per cent of mucosal stages of Trichonema spp., while doses of 60 mg/kg removed 83 per cent and 89 per cent of the migrating larvae of S. vulgaris and S. edentatus respectively.


Two dopamine receptors: supportive evidence with the rat rotational model. Di-isobutyryl apomorphine (5 mg/kg i.p.) and lergotrile (5 mg/kg i.p.) produce long lasting turning behaviour in rats with 6-hydroxydopamine lesions at the level of the substantia nigra. Haloperidol (1.5 and 3.0 mg/kg p.o.) blocks the rotational behaviour due to the apomorphine ester but has no effect on lergotrile turning. Clozapine (10 and 20 mg/kg p.o.) blocks the lergotrile turning but stimulates the rotational behaviour produced by the apomorphine ester. The results support the concept of two anatomically separate dopamine receptors and their relevance to the study of antipsychotic activity is discussed.


Initiation of RNA synthesis in isolated nuclei. Ribonucleotide triphosphates, labeled at the beta position, were synthesized and used directly to quantify RNA chain initiation in nuclei isolated from Friend cells grown in tissue culture. At the optimal salt concentration, low-molecular-weight RNAs (4-5 S) synthesized by RNA polymerase III were the predominant species initiated. Less than 5% of the molecules were initiated by polymerase II. We calculate that 50-80% of the small RNA molecules synthesized in vitro were also initiated in vitro. Assuming that a substantial fraction of the nuclei were active in vitro, the number of 4-5 S RNA molecules initiated per nucleus was about 100 molecules/min.


Changes in striatal dopamine metabolism during precipitated morphine withdrawal. Precipitation of withdrawal in morphine tolerant/dependent rats by either naloxone or the partial agonist ZK 48491 caused a significant increase in the contration of striatal DA, which persisted for at least 1 h. During the same time the probenecid-induced accumulation of HVA and DOPAC was reduced in the striatum in relation to probenecid-treated tolerant/dependent controls. 20 min after precipitation of withdrawal by naloxone, the striatal concentration of 3-methoxytyramine was decreased by about 40%, while the activity of the DA metabolizing enzymes, MAO and COMT, remained unchanged. Naloxone-precipitated withdrawal was, further, found to delay the depletion of striatal DA caused by inhibition of synthesis 90 min after alpha-methyl-p-tyrosine treatment. All these results provide evidence for a decreased release of DA from the striatum during precipitated morphine withdrawal.


Effects of carmine and carminic acid on embryonic tissue cell cultures. The biological reaction to carmine and carminic acid at cellular level on 'in vitro' cultures was tested and significant variables were controlled. Results suggested that proliferation and metabolism of these cultures were not affected by the 2 stains.


Binding of indole-acetic acid to cytosol proteins of embryonic chicken liver. 3H-Indole acetic acid bound to cytosol proteins of embryonic chicken liver in vitro to an average capacity of at least 20 pmoles/mg binding protein. The auxin-binding protein complexes could be resolved into 4 major zones by anion exchange chromatography; they sedimented at 3-7 S in sucrose density gradients, and were also heterogenous in agarose gel electrophoresis.


How to view stereoscopic pictures of crystal structures and molecular models. Methods are presented for practicing the viewing of stereoscopic pictures of crystal structures and molecular models without optical aids.


Effect of antiblastokinin on rabbit pregnancy. On different days following coitus, i.e., during the first stages of pregnancy, adult female rabbits were treated with antiblastokinin obtained from a chicken. A high prenatal mortality was observed and a reduction of the weight in the stillborn of the rabbits treated. The antiblastokinin produces evident biological effect about the 4th day following coitus, during the period of maximum production of blastokinin by the endometrium in vivo.


Desmosomal abnormalities in the liver of methotrexate-treated psoriatics. Electron microscopy of the liver of methotrexate-treated psoriatic patients revealed junctional abnormalities consisting of detachment of desmosomal plaques between hepatocytes. Mitochondria anchoring desmosomal microfilaments were frequently noted.


Effects of the antibiotic netropsin on mouse ascites tumour chromosomes in vitro. The antibiotic netropsin was found to induce an increase of the aberration frequency of up to 10% and a decondensation and elongation of centromeric regions of the chromosomes in mouse ascites tumour cells cultivated in vitro.


Effect of cholinesterase inhibitors on differentiation of cultured sympathicoblasts. The 2 cholinesterase inhibitors, eserine and BW 284C 51, inhibited the nerve fibre growth and differentiation of immature sympatheticoblasts at low concentrations. The effect was nerve cell-specific and appeared in the course of the in vitro development, indicating that cholinesterases might play an important role in the early differentiation of sympathicoblasts.


Androgen-binding activity in the human testis and epididymis. The quantitation of androgen-binding activity in human testes, epididymides, and serum is reported and compared with that of similar tissue in the rat. By using steady-state polyacrylamide gel electrophoresis and a tritiated 0.5 nM testosterone label, values for specific binding activity, reported as picomoles per milligram of protein, were found to be 0.21 +/- 0.01 in normal human, epididymides, 0.03 in testes, and 0.44 +/- 0.07 in human serum. Tissues from men receiving exogenous estrogen were also similarly analyzed and found to possess significantly higher androgen-binding activity: 4.10 +/- 0.48 pmoles/mg of protein (epididymides), 4.01 (testes), and 5.63 +/- 1.15 (serum). Since man possesses a sex steroid-binding globulin, attempts were made to differentiate Ion-exchange chromatography, steroid specificity, and dissociation half-time determinations were suggestive but did not conclusively demonstrate the existence of two distinctly different androgen transport macromolecules in man.


[Regional redistribution of blood during emotional excitation in rats]. Shifts in general circulation (GC) and changes of relative blood volume in 46 vascular areas were studied in unanesthetized rats closed in a small cell for 10 min, 1, 3, 4.5 hrs as well as in rats tied with their backs down for 1 hr. The GC increased in the latter case. Regional redistribution of the blood volume was not related to the position of the body but its intensity was related to the degree of emotional excitation. All the tests reduced the relative blood volume in a number of abdominal organs and in small pelvis; in muscles and bones of limbs, abdomen and pelvis; in skin of the breast and hindlimbs. The percent blood volume increase occurred in the brain, heart, lungs, muscles and bones of the head, neck, and breast. Regional blood redistribution in rats tied to a bar with their backs down for 4 hrs involved a decrease of the relative blood volume in the skin, muscles and bones, in a number or organs of the small pelvis and abdomen, while the volume increased in the liver, heart, and lungs.


[2 cases of subaortic stenosis with subvalvular diaphragm complicated by postendocarditis aortic insufficiency]. Two cases of patients with discrete subaortic stenosis complicated by aortic regurgitation resulting from the sequelae of bacterial endocarditis and treated by valvular replacement are described. A correct preoperative evaluation was due to echocardiographic data and pressure curves; left ventriculography did not show any evidence of subaortic obstruction in any of the cases. In one patient, bacterial endocarditis involved mitral valve too, which was replaced by an artificial prostheses. Bacterial endocarditis is very rare in patients with discrete subaortic stenosis, but it must be regarded as a factor complicating the natural history of cardiopathy.


Influences of ulcerogenic stimulations in the stomach with the clamping ulcers in rats. Ulcerogenic stimulations such as histamine and restraint water immersion stress were given to rats after the gastric ulcers were made by the clamping method. Development of the stress and histamine ulcers were distinctly inhibited after the clamping ulcers were made, and this inhibition reached a maximum at the 7th clamping ulcer day and lowered gradually thereafter. In control rats, the stress and histamine ulcers usually develop in the lesser curvature, but in animals with the clamping ulcers they develop in the greater curvature where the clamping ulcer was made. The histological changes of the stress ulcers in the stomach with the clamping ulcer were slighter compared with those of the control (without clamping ulcers) rats. Furthermore, the relationships between the development of the stress ulcers and the changes of the uronic acid content in the stomach were observed. The uronic acid content increased after the clamping ulcer was made and reached a maximum at the 7th clamping ulcer day. Thereafter it began to decrease and returned to the normal level. From the results of the studies, it was strongly suggested that the mechanism of inhibition to development of the stress ulcers might be attributed to an increase of acid mucopolysaccharide in the gastric wall after the clamping ulcer was induced.


[Hybridization of Chinese hamster cells sensitive to ultraviolet irradiation]. Resistance to UV-light was studied in two UV-sensitive aneuploid Chinese hamster cell clones to different origin and degree of sensitivity, their respective polyploids and somatic cell hybrids. The karyotype of the parental clones, cell hybrids and polyploids was analyzed in parallel. A great variability of karyotypes was detected in hybrid cells. Serial cultivation of hybrids was accompanied by chromosome loss. Soon after fusion the hybrid clones proved to be more resistant to UV than the parental sensitive cells. However, their sensitivity increased with passages. The comparison of UV-sensitivity with data on karyotype analysis allowed to assume that the increase in sensitivity was correlated with the loss of particular chromosomes or chromosome regions. The results obtained indicated the existence of a polygenic control of UV-sensitivity, the multiple genes being assigned to different chromosomes. A reverse effect of ploidy was detected, i.e. a decrease in the resistance to the lethal action of UV-light in polyploids as compared to the parental clones.


The metabolism of caffeine by a Pseudomonas putida strain. 1) A bacterium capable of growing aerobically with caffeine (1,3,7-trimethylxanthine) as sole source of carbon and nitrogen was isolated from soil. The morphological and physiological characteristics of the bacterium were examined. The organism was identified as a strain of Pseudomonas putida and is referred to as Pseudomonas putida C1. 15 additional caffeine-degrading bacteria were isolated, and all of them were also identified as Pseudomonas putida strains. The properties of the isolates are discussed in comparison with 6 Pseudomonas putida strains of the American Type Culture Collection. 2) The degradation of caffeine by Pseudomonas putida C1 was investigated; the following 14 metabolites were identified: 3,7-dimethylxanthine (theobromine), 1,7-dimethylxanthine, 7-methylxanthine, xanthine, 3,7-dimethyluric acid, 1,7-dimethyluric acid, 7-methyluric acid, uric acid, allantoin, allantoic acid, ureidoglycolic acid, glyoxylic acid, urea, and formaldehyde. Formaldehyde has been demonstrated to be the product of oxidative N-demethylation mediated by an inducible demethylase. A pathway of caffeine degradation is proposed.


Are globin coding sequences expressed in nuclear RNA of chick embryo brain? The content of globin coding sequences in nuclear RNA from chicken embryo red blood cells and chicken embryo brain was determined by hybridization with globin cDNA to be 0.27% for embryonic red blood cells and 0.001% for embryo brain, i.e. 0.37% of globin coding sequences relative to embryonic red blood cells. By transfusion of [3H]uridine labelled red blood cells it could be shown that the brain nuclear RNA preparation was contaminated by 0.41% RNA originating from blood cells. As this value is in the same range as the hybridization rate there is no evidence for globin transcripts in brain nuclear RNA. It has also not been possible to detect a short-lived globin mRNA precursor in pulse labelled brain nuclear RNA. In further experiments the RNAs were translated in an Ehrlich ascites cell-free system. No globin synthesis could be observed with brain RNA.