Histological and ultrastructural study on the subcommissural organ of Bradypus tridactylus.

The subcommissural organ (SCO) of the Bradypus tridactylus was studied from the histological and ultrastructural point of view. The methods of GOMORI (alum-chromic-hematoxylin-phloxine), AZAN (M. Heidenhain), KLUVER and BARRERA (for nerve cells and myelinated fibers) and silver impregnation according to PALMGREN and CAJAL-DE CASTRO, were used. The material prepared for ultrastructural analize was submitted to the sequence of gluteraldehyde, osmium tetroxide, uranyl acetate and lead citrate. In function of the secretory phenomenon, a systematic study is made of different hystological and ultrastructural characteristics of the SCO and hypendymal rosettes, which were considered in the B. tridactylus as a morphofunctional unit. Fine vascular and nerve networks were observed, by the basal parts of the compact tubule alveolar system (hypendymal rosettes) which composes part of the SCO. Nerve fibers were seen between its cells reaching even up to the lumen, and they may end there in bulbar dilatations which seem to show secretory characteristics at the ultrastructural examination.


An autoradiographic study of long-term follicular development in the cyclic hamster.

The time required for ovarian follicular development in the cyclic hamster was determined by marking follicles with [3H]-thymidine on day 1 of the estrous cycle (day of ovulation) and by following their fate over the next three cycles by autoradiography. A model of follicular development was formulated, based on the total number of follicles and the changing percentage of labelled follicles with time. It was estimated that 20 days elapsed before a preantral follicle with two to three layers of granulosa cells (stage 1) was able to ovulate. Follicles spent eight days in stage 1 and an additional eight days in stage 2 (4-5 granulosa layers). Preantral follicles entering stage 3 (6-7 granulosa layers) and 4 (greater than 8 layers) on day 1 of the cycle ovulated four days later. Approximately 30 follicles per ovary entered and left stage 1 each cycle, representing a 50% turnover. Of the follicles leaving stage 1, about 50% were transformed into stage 2 follicles; the remaining ones underwent atresia. Stage 2 follicles also had a 50% turnover each cycle. Half of the stage 2 follicles underwent atresia; the remainder developed into stages 3 and 4 follicles. Fifty percent of the stages 3 and 4 follicles present on day 1 of the cycle ovulated at the next estrus with the remainder undergoing atresia.


Plasma cortisol in glaucoma.

Plasma cortisol level and intraocular tension were determined diurnally in 40 normal and 40 glaucomatous patients at 8 AM and 8 PM. It was observed that (1) plasma cortisol level was higher in glaucomatous patients at both the periods; (2) the diurnal variation in plasma cortisol was parallel to diurnal variation in intraocular tension in both glaucomatous and normal persons; and (3) the diurnal variation in plasma cortisol level also was more marked in glaucomatous patients as compared to normal persons.


[Gongylonema pithyusensis n. sp. (Nematoda: Spiruridae), a parasite of the oesophagus of the dormouse Eliomys quercinus ophiusae Thomas, 1925 (Rodentia: Gliridae) in Formentera (Balearic Islands) (author's transl)].

A new Nematode, Gongylonema pithyusensis n. sp., parasitizing the oesophagus mucosa of the dormouse Eliomys quercinus ophiusae Thomas, 1925 in the balearic island of Formentera (Spain) is described. The male of the new species is characterized by the size of spicules and gubernaculum; the female by the morphology of the tail, the posteriour situation of the vulva and the dimensions of the eggs. It is the only Gongylonema species to be known parasitizing Rodents of the family Gliridae.


Effect of prostaglandins F2alpha and E2 on milk ejection, blood pressure and mammary artery blood flow in the cow.

Injections of prostaglandins F2alpha (threshold dose: 1 to 5 microgram) and E2 (threshold dose: 1000 to 300 microgram) in the jugular vein or the carotid artery induced milk ejection in lactating cows after a relatively long latency period. In several cases with PGF2alpha and in one case with PGE2, milk ejection was accompanied by a simultaneous increase in blood flow through the mammary artery. Both routes of administration showed the same response. It is suggested that the effect of the prostaglandins on milk ejection is indirect, possibly secondary to a release of oxytocin from the neurohypophysis.


Mitral annular calcification. Clinical and echocardiographic correlations.

Clinical, echocardiographic, and radiographic findings in 41 patients with mital annular calcification are presented. Clinical diagnosis included rheumatic heart disease (32%), noninflammatory calcific disease (34%), and chronic renal failure (32%). Mitral annular calcification is recognized echocardiographically as a dense echo band beneath the posterior mitral leaflet with motion paralleling that of the free left ventricular wall. Only two patients of 41 (5%) with mitral annular calcification had calcification on routine x-ray films, and six of 16 (38%) on cinefluoroscopy. The left atrial size is enlarged, and the mitral leaflets are thickened, with reduced motion (D-E) in rheumatic heart disease, but the leaflet motion is well preserved in the nonrheumatic patients. The association of mitral annular calcification with idiopathic hypertrophic subaortic stenosis, chronic renal failure, and bacterial endocarditis is discussed. Echocardiography appears to be a sensitive method of detecting mitral annular calcification and may be helpful in differentiating rheumatic and nonrheumatic etiologies.


Infective endocarditis caused by Streptococcus mutans. A complication of idiopathic hypertrophic subaortic stenosis.

Three patients with endocarditis caused by Streptococcus mutans were seen during a six-month period. All had clinical features of subacute bacterial endocarditis, including fever, heart murmurs, and positive blood cultures. One had underlying aortic insufficiency and two had idiopathic hypertrophic subaortic stenosis. All patients were treated with parenteral antibiotics and were cured. Streptococcus mutans is a pleomorphic, microaerophilic organism that is associated with dental caries and plaque. Differentiation of S mutans from enterococcal endocarditis is important because the former condition can be treated for a shorter period of time with penicillin alone, without the addition of aminoglycoside antibiotics.


[Some comments on the biological properties of calcitonin: an eventual new therapeutic utilization (author's transl)].

The calcitonin-adrenal antagonism brings the authors to propose a new therapeutic use of calcitonin. That hormone could minimize the effects of externe stimulations and its use as sedative is proposed.


[Ultrastructure of the lymphatic capillaries of the diaphragm and their interrelationship with peritoneal mesothelium].

In 28 mature rats, ultrastructure of the diaphragmal lymphatic capillaries and mesothelial tegmen was studied. Interrelation of their cellular elements and possible ways of metabolic absorbtion from the abdomen were clarified. The diaphragmal lymphatic capillaries were demonstrated to situate under mesothelial cells and they are separated by a thin layer of longitudial collagenic fibers. As cross section of the lymphatic capillary demonstrates, the capillary wall consists of 3--6 endothelial cells with a set of organelles, among which are: mitochondria, Golgi's complex, some pinocytotic vesicles.


Evolution of neovascularization in sickle cell retinopathy. A prospective fluorescein angiographic study.

With prospective observations we have documented by fluorescein angiography the development of retinal neovascularization in the eyes of patients with sickle cell-hemoglobin C disease (hemoglobin SC). In each patient, arteriolarvenular (AV) anastomoses occurred before the development of neovascular sea fans. To our knowledge, this report constitutes the first such documentation in the literature. New sea fans from AV anastomoses appeared throughout an average interval of 18 months (range, 8 to 36 months) in approximately 21% of young adults with hemoglobin SC. Each year approximately 14% of young patients with hemoglobin SC may develop retinal neovascularization.


Concentrations of luteinizing hormone in ovariectomized ewes and the effect of disturbance, starvation and vascular cannulation.

Neither routine experimental procedures, including vascular cannulation and venipuncture, nor starvation or disturbance caused significant fluctuations in luteinizing hormone (LH) concentrations in ovariectomized ewes. Samples obtained at 5-min intervals revealed episodic LH releases occurring every 10-35 min. Hourly sampling over 3 days revealed large LH variations but no distinct diurnal variation.


Studies of the antigenicity and immunogenicity of bromelain-pretreated red blood cells.

The effects of the proteolytic enzyme bromelain (Br) on the antigenicity and immunogenicity of sheep and mouse red blood cells (RBC) have been investigated. The results presented support the previous claim that there are antigens present on Br RBC that are not present in an exposed form on untreated RBC and that Br RBC have lost some of the antigens present on the surface of normal RBC. The susceptibility of Br RBC to osmotic lysis was very similar to that of normal RBC, implying that the modified RBC were not more fragile than normal RBC. Injection of mice with Br mouse-RBC did not increase the unusually high "background" number of cells producing IgM antibodies against Br mouse-RBC and mice did not mount delayed-type hypersensitivity reactions against Br mouse-RBC, either before or after sensitizing injections of Br mouse-RBC. However, mouse-RBC and Br mouse-RBC elicited similar antibody responses in rabbits and guinea pigs. Although mice appeared unresponsive to Br mouse-RBC injections, delayed-type hypersensitivity responses and antibody production in primary and secondary responses were of similar levels irrespective of whether sheep-RBC or Br sheep-RBC were used as immunogens. From these studies it appears that mice have B-cells producing antibodies against the "new" antigens on Br mouse-RBC, but there are no T-cells that respond to these antigens by way of "helper" activity in antibody production or by way of cell-mediated immune reactions.


Eumycotic mycetoma due to Drechslera rostrata infection in a cow.

A case of eumycotic mycetoma with lesions in the skin, nasal cavity and lymph nodes of a cow is reported. The case history, clinical course, macroscopic and microscopic appearance of lesions, and fungal isolation are described. Drechslera rostrata was isolated from pathological tissue and Koch's postulates were satisfied on mouse inoculation. This is the first infection of its type recorded in Australia and features contrasting with cases reported in other countries are discussed.


Mycotic nasal granuloma in cattle.

Seven cases of eosinophilic granulomas containing fungal chlamydospores and short septate hyphae in the submucosa of the anterior nasal cavity were described from Queensland beef cattle during 1966-75. Drechslera rostrata was isolated from 2 cases and unidentified dematiaceous fungi from 2 others.


[Absence of cutaneous phenomena of delayed hypersensitivity in the use of zymosan as adjuvant in comparison with Freund's complete adjuvant in autoimmune experimental orchitis].

In previous researches some of the Authors proved that Zymosan acts as adjuvant in determining the experimental autoimmune aspermatogenesis, in absence of skin reactivity of delayed type. The aim of the present investigations was to ascertain if Zymosan possess some components analogous to those of mycobacteria, which can be considered responsible in determining hypersensitivity reactions of delayed type. The results, obtained also in cross experiments, don't confirm this hypothesis.


Carbon-13 nuclear magnetic resonance studies of the binding of selectively 13C-enriched oxytocins to the neurophypophyseal protein, bovine neurophysin II.

Complex formation between bovine neurophysin II and oxytocin molecules containing 85% 13C enrichment in specific amino acid residues was studied using 13C nuclear magnetic resonance spectroscopy. Chemical shift and relaxation time values of the analogue [13C-Leu3]oxytocin, [13C-Gly9]oxytocin, and the doubly labeled [13C-Ile3 Gly9]oxytocin were obtained for the hormones in the absence and presence of neurophysin. The results showed that certain 13C nuclear magnetic resonance parameters of residue 3 but not of residue 9 of oxytocin are altered upon binding to neurophysin. These observations suggest that residue 3 but not residue 9 is involved in the protein-hormone interaction and they demonstrate the general applicability of selective 13C enrichment for the study of peptide-protein interactions.


Surface potential effects on metal ion binding to phosphatidylcholine membranes 31P NMR study of lanthanide and calcium ion binding to egg-yolk lecithin vesicles.

31P NMR of phosphatidylcholine (lecithin) from egg-yolk in sonicated vesicles has been measured in the presence of various ions. Addition of Ln3+ or Ca2+ shifted the 31P resonance of the phosphate groups of the outer surface of the vesicles. These shifts were measured at varied lanthanide or Ca2+  concentration at different ionic strengths obtained by addition of NaCl. The shifts induced by Tb3+ and Ca2+ have been analyzed using the theory of the diffuse double layer. Corrections were introduced for the effect of the ionic strength on the activities of the ions. The binding efficiency is shown to be controlled by the electrostatic potential produced by the bound cations at the membrane surface. This potential is slightly modified due to weak chloride binding. Binding constants have been derived.


Effects of wheat germ agglutinin on membrane transport.

(1) Low concentrations of wheat germ agglutinin are cytotoxic toward several tissue culture lines, including Chinese hamster ovary cells, Swiss 3T3 cells, mouse L cells and baby hamster kidney cells. The LD50 ranged from 1 to 5 microgram wheat germ agglutinin per ml. Similar concentrations of the lectin inhibited the transport of the non-utilizable amino acids alpha-aminoisobutyric acid and cycloleucine and inhibited the uptake of thymidine. In contrast, 2-deoxy-D-glucose uptake was not altered and colchicine uptake was enhanced. (2) The inhibition of alpha-aminoisobutyric acid uptake occurred within minutes after lectin addition and was maximal by 1 h. Maximal inhibition ranged from 50 to 70% of control values. Studies of the kinetics of the uptake demonstrated that wheat germ agglutinin decreased the V of the uptake by 70% without affecting the apparent Km. Ovomucoid, a haptene inhibitor of wheat germ agglutinin-binding to cell surface receptors, prevented the wheat germ agglutinin-induced inhibition of alpha-aminoisobutyric acid transport. Three other lectins (Concanavalin A, Phaseolus vulgaris E-phytohemagglutinin and L-phytohemagglutinin) inhibited the uptake by 20% or less at doses up to 50 microgram/ml. (3) We propose that the cytotoxicity of wheat germ agglutinin probably results in part, if not totally, from membrane alterations which impair multiple membrane transport systems.


Does the non-saturable cell entry apply to the charge-free form of amino acids?

The slow cellular entry shown by neutral alpha-amino acids at very high concentrations appears not to arise from diffusion of the totally uncharged species through the plasma membrane of the Ehrlich cell, judging from a similarity of the rates observed for the two conformational isomers of 1-amino-2-hydroxy-cyclohexane-carboxylic acid. One of these isomers provides in neutral solution 4 times as large a proportion of the charge-free species as the other, and 5 times the proportion calculated for alanine.


Electrophysiology of phagocytic membranes. I. Potassium-dependent slow membrane hyperpolarizations in mice macrophages.

Electrophysiological properties of activated mouse macrophages cultured in vitro were studied using microelectrode techniques. In a high percentage of the individual cells analysed a slow hyperpolarization (SH) was observed with a concomitant decrease (2--4 times) of the input resistance. Increasing doses of tetraethyl ammonium progressively reduce the amplitude of the SH and at a concentration of 15 mM complete blockade of the phenomena is observed. Valinomycin, at a concentration of 10(-7) M produces rapid and permanent hyperpolarization, with a shift in the membrane potential to about --50 mV. These data strongly support the previously proposed hypothesis that the development of SH is due to an increase in the membrane permeability to potassium ions.


Calcium metabolism in Ehrlich Ascites tumour cells.

Ehrlich ascites tumour cells are able, under the proper experimental conditions, to extrude a substantial amount of Ca2+ from the intracellular space. The Ca2+ extrusion mechanism, probably located at the plasma membrane level, appears to be similar to that found in red blood cells. It is energy-dependent and both respiration and glycolysis are able to drive it. The use of some inhibitors and uncouplers, besides showing that this activity is different from that linked to the mitochondrial Ca2+ pump which acts in the opposite direction, proposes some speculations on the energy compartmentation in the Ehrlich ascites tumour cells.


Developmental regulation of nuclear glycosyl transfer in Cictyostelium discoideum.

Glycosyltransferases are active in isolated cell nuclei of Dictyostelium discoideum. Since the biological role of these nuclear enzymes is unknown we assayed their activity in nuclear fractions from vegetatively growing and differentiating amoebae. The nuclear N-acetylglucosaminyl transfer is highest in the vegetative stage and shortly after induction of differentiation, then declines gradually. This result is discussed with respect to a possible role for glycoproteins and mucopolysaccharides in the structure or in the transcriptional and replicative functions of the genome.


Isolation of the porcine ileal intrinsic factor receptor by sequential affinity chromatography.

An extract containing solubilised receptor was passed through four columns containing Sepharose to which had been covalently coupled anti-cobalophilin IgG, vitamin B-12-intrinsic factor, vitamin B-12, and free intrinsic factor, respectively. Following a wash the receptor was eluted with EDTA, then residual Triton X-100 micelles and vitamin B-12-intrinsic factor were removed by Sephadex G-200 filtration. The receptor was purified 84 000-fold, sodium dodecyl sulphate electrophoresis indicated two subunits and gel filtration of its vitamin B-12-inttrinsic factor complex resolved it into two molecular species.


Differential effect of pre- and postnatal litter size reduction on body weight and development of stress response in the rat.

The postnatal development of pups belonging to the small, prenatally reduced litters (PRN) was compared to that of pups of the postnatally reduced litters (PST) and to that of pups of intact, normal-size litters (CON). Body adrenal and pituitary weight gain as well as maturation of the pituitary-adrenal system were enhanced in the PRN and PST pups as compared to the CON pups in the 3 postnatal weeks. However, significant differences in some data between the PRN and PST pups were found, indicating the more pronounced beneficial effect of the prenatal reduction of litter size. The postnatal litter size reduction abolished the weight gain of the testes. The prenatal reduction of litter size resulted in a similar, although less pronounced negative effect.


[Hypnogenic action of a modulated electromagnetic field].

Behavioristic and electroencephalographic changes in rats under long-lasting modulated electromagnetic field (frequency of 40 MHz, modulation frequency 50 Hz, intensity 100--120 V/m) were studied. Some phasic disorders in the conditioned feeding and defense reactions were observed, including cataleptic state as a result of the action of modulated electromagnetic field. These behavioristic disorders are determined by the changes in the normal cortico-subcortical relationships.


Activation of bovine factor VII by hageman factor fragments.

During the early events of coagulation of human blood by the intrinsic pathway, factor XII is activated to a form which can activate factor XI, and is proteolytically fragmented to smaller species (30,000 daltons and 70,000 daltons) which have lost most of the ability to activate factor XI but which can activate prekallikrein rapidly. The effect of these fragments on factor VII was studied. It was found that these Hageman factor fragments promoted rapid proteolysis of one-chain factor VII to a more active two-chain form. The amino-terminal sequences of the chains of activated factor VII were found to be Ala-Asx-Gly- and Ile-Val-Gly-, the same as were earlier observed after activation of factor VII by activated factor X. This finding indicates that initiation of coagulation by the intrinsic pathway also primes the extrinsic pathway.


Erythroid cell growth from normal and W/WV murine bone marrow on macrophage-coated membranes.

Cellulose acetate membranes (CAM) placed in the peritoneal cavity of mice develop a macrophage layer capable of supporting in vivo hematopoietic colonies from intraperitoneally injected bone marrow cells. Modifications allowing for routine morphologic identification of colonies showed that both erythrocytic (E) and granulocytic (G) colonies occur with a consistent E:G ratio of 0.19 +/- 0.037. Stimulating recipients by bleeding or phenylhydrazine injection did not produce a significant change in the total number of colonies and a reduction in granulocytic colonies so that the E:G ratio significnatly increased. Hypertransfusion of donor animals had no effect on the number of erythroid colonies that grew on CAM of average recipients. The total colony-forming ability of bone marrow cells from genetically anemic W/WV mice was found not to differ from that of normal +/+ littermates; however, the E:G ratio of W/WV marrow in bled recipients was significantly lower (p less than 0.01) then that of +/+ marrow. These studies suggest that a CAM system supports an erythroid progenitor which is not affected by hypotransfusion of the donor animal, yet is dependent upon erythropoietin for colony formation, and that it is defective in the W/WV mouse.


Cellular fluxes of vitamin B12.

Incorporation of vitamin B12 into L1210 cells requires the protein binder transcobalamin II (TCII). The process is saturable, follows Michaelis-Menten kinetics (Km = 2.5 X 10(-9) M at 37 degrees C), is both temperature and calcium (K50 - 1 X 10(-6) M) dependent, and is inhibited by apo-TCII, indicating the presence of a TCII specific receptor on the cell membrane. B12 also leaves the cell by a calcium-independent pathway bound to either TCII or to a protein with chromatographic properties similar to those of TCIII. Since intact TCII-B12 can be found  in the cytosol and can promote B12 uptake by mitochondria, it is proposed that the B12 released from the cell bound to the TCIII-like protein is derived by mitochondrial processing of incorporated TCII-B12. The slower time course of release of the latter B12 is consistent with this postulate.


Diurnal plasma free fatty acid profiles in normal and diabetic pregnancies.

Diurnal plasma free fatty acid (FFA) and glucose concentrations were measured in 23 normal women, 13 chemical diabetics, and 11 insulin-dependent diabetics in late pregnancy. The mean diurnal FFA value was lowest in the insulin-requiring diabetics and highest in the chemical diabetics. Although no association was found between percentile birthweight and maternal mean fasting or diurnal FFA, there was a positive correlation between percentile birthweight and both mean maternal fasting plasma glucose and mean diurnal glucose in normal and chemical diabetic women. These observations support the view that glucose is the major substrate used by the fetus, but birthweight may be influenced more by the total substrate crossing the placenta than by maternal levels of either glucose or FFA alone.


Effects on germ-free and conventional quail of substituting isolated soya protein for free amino acids in a purified diet.

1. Germ-free and conventional Japanese quail reared on a diet in which the nitrogen was supplied solely in the form of free amino acids grew well, but growth was significantly better in both environments when some of the amino acids were replaced by an equivalent quantity of isolated soya protein. 2. Although the small intestine was shorter in the birds given the soya protein supplement, its weight per unit of length was greater; histological examination suggested that its circumference and wall thickness were both increased. 3. Results of a small experiment with conventional quail given the diets marked with 14C-polyethylene glycol indicated that the free amino acid diet passed through the gut more rapidly than the supplemented diet. 4. It was concluded that the growth-promoting effect of the isolated soya protein was unrelated to the activities of the gut microflora but might be due to changes in the digestive and absorptive capacities of the small intestine induced by the different physical nature of the supplemented diet.


Formiminotransferase-cyclodeaminase from porcine liver. A sulfhydryl essential for the deaminase activity of the bifunctional enzyme.

Reaction of the bifunctional enzyme formiminoglutamate:tetrahydrofolate formiminotransferase (EC 2.1.2.5) - formiminotetrahydrofolate cyclodeaminase (EC 4.3.1.4) with the sulfhydryl reagent 5,5'-dithiobis (2-nitrobenzoic acid) selectively inactivates the cyclodeaminase. Loss of activity correlates with the modification of two sulfhydryl groups per subunit. The inhibitor folic acid reduces the rates of inactivation and sulfhydryl modification, and protection experiments demonstrate that only one of the two sulfhydryls modified is important for enzyme activity. The results indicate the presence of a cyclodeaminase site on each polypeptide, assuming one sulfhydryl per site, in agreement with a quaternary structure containing identical polypeptides. Modification does not cause dissociation of the enzyme and is reversible with dithiothreitol.


Comparison of colony morphology, salt tolerance, and effectiveness in Rhizobium japonicum.

Four strains of Rhizobium japonicum, two of which produce slimy and non-slimy colony types and two others which produce large and small colony types, were isolated and cloned. All were infective and nodulated Lee soybean host plants. Each colony type was characterized as to its salt sensitivity to Na+ and K+ ions, relative level of symbiotic nitrogen fixation, and relative level of free-living nitrogen fixation. Growth studies performed in the presence of salts demonstrated that the non-slimy or small colony types were sensitive to salt with significantly depressed growth rates and cell yields. Growth rates and cell yields of slimy, large, colony types were relatively unaffected by salt. Both symbiotic and free-living (non-associative) nitrogen fixation analyses (by acetylene reduction) revealed that the non-slimy, small colonies were significantly more effective than slimy, large colonies.


Contrasting effects of rabbit and human platelets on chikungunya virus infectivity.

Chikungunya virus infectivity was markedly stabilized in the presence of washed suspensions of human platelets but rapidly disappeared in similar preparations of rabbit platelets. Supernatant fluids collected from human platelets had some stabilizing effect on chikungunya virus over a 6-day incubation period at 37 degrees C. Rabbit platelet supernatant fluid had no virus-stabilizing effect, nor did it demonstrate any capacity to inactivate virus as compared to whole rabbit platelet preparations. Thin-section election microscopy demonstrated that chikungunya virus formed an associated with human platelets by becoming entrapped in platelet aggregates; during this process some of the platelets appeared to have undergone degranulation and lysis. Rabbit platelets exposed to chikungunya virus for 24 h demonstrated a considerable amount of platelet degranulation and lysis but virus was not visualized either in association with platelet membranes or within phagocytic vacuoles in the platelet cytoplasm. Human platelets, which appear to be more stable under these incubation conditions, may protect chikungunya virus infectivity from heat inactivation by surrounding viruses with large platelet aggregates whereas rabbit platelets, which appear to be more fragile, do not afford this type of protection. Thus, chikungunya virus in the presence of rabbit platelets may become inactivated by heat or may become bound irreversibly to membranes in such a fashion that infectivity assay and electron microscopy techniques may prove to be too insensitive for detection of virus.


Deoxyribonucleic acid base composition of certain species of the genus Bacteroides.

The moles percent guanine plus cytosine content of the DNA (% G + C) of 15 Bacteroides strains representing six species was determined. One group, including three strains of Bacteroides ruminicola, two strains of B. melaninogenicus, two strains of B. succinogenes, and one strain of B. oralis (JI), had a % G + C of 47.6--50.3 and a second group including two strains of B. amylophilus, four strains of B. fragilis, and one strain of B. succinogenes had a lower % G + C of 40.3--42.7. The taxonomic relationships among these bacteroides species were discussed.


Characterization and radiobiologic parameters of medulloblastoma in vitro.

Cultured cells from human medulloblastomas were demonstrated to have distinctive features and a "glial" potential from the morphological and ultrastructural parameters. The in vitro radiobiological survival curve parameters of cultured human medulloblastoma and non-malignant skin fibroblasts obtained under the same conditions of exponential growth were similar. In vitro conditions may underestimate the shoulder region of the survival curve. Since the shoulder region is the most important in clinical fractionated radiotherapy, multiple dose experiments might be useful in examining this portion of the survival curve more closely.


Ultrastructure of gonadoblastoma and disgerminoma (seminoma) in a patient with XY gonadal dysgenesis.

The gross and fine morphology of gonadoblastoma and disgerminoma in a young phenotypic female with male karyotype and gonadal dysgenesis is described. Both the rudimentary gonads contain gonadoblastomas on the left side in association with a disgerminoma. In the light microscope the gonadoblastomas show peculiar hyaline masses, surrounded by Sertoli-like cells. In some areas a direct continuation between hyaline masses and basement membranes of round nests or large anomalous seminiferous tubules is obvious. At the ultrastructural level the hyaline masses consist of densely packed, largely concentric membranes closely similar to basement membranes or basal laminae of seminiferous tubules. These observations and the finding of transitional zones with gonadoblastoma elements in enlarged tubular structures suggest that gonadoblastomas originate inside tubules of dysgenetic testes. The peculiar hyaline masses with their concentric membranes might reflect hyperplasia and death of the Sertoli cells, each new generation of cells producing a basement membrane outside the preceding one.


Lymphangiography in patients with ovarian epithelial cancer: an evaluation of 289 consecutive cases.

From January, 1973, to June, 1976, 226 patients with palpable ovarian masses were evaluated preoperatively by lymphography. Histology showed 166 cases of malignant epithelial tumors, 26 benign tumors, and 34 malignant special tumors (not included in this report). Furthermore, the group of patients included 99 recurrences of ovarian epithelial cancer and 24 patients who underwent restaging diagnostic procedures without clinical evidence of disease. Lymphography was negative in all patients with benign tumors. In the 289 cases of epithelial cancer, lymphangiography gave evidence of nodal metastases in 88 (30%). When the histological subtype was considered, the highest incidence of metastases was in undifferentiated carcinoma (50%) and the lowest, in mesonephroid carcinoma (14%). According to the stage before lymphography, nodal metastases were found in 8% of Stage I, 0% of Stage II, 29% of Stage III, and 53% of Stage IV cases. The incidence of metastases was 46% in patients studied for recurrent disease and 17% in patients studied for restaging. Fifty-four percent of patients had metastases only in the pelvic nodes and 18% only in the para-aortic chains; in 28% both chains were involved simultaneously. Bilateral involvement was found in 63% of the positive cases. Retroperitoneal node biopsies were performed in 68 patients (36%). The radiologic/histologic correlation was 100% in the lymphangiographically positive cases; 81% in the negative cases, with nine false-negative reports; and 87% in all cases.


Fibrosarcoma in infants and children: a review of 110 cases.

This retrospective study summarizes the clinicopathologic findings in 110 children with fibrosarcoma. Seventy cases were selected from previous reports and 40 were from our files. The histologic criteria used for diagnosis are the same as those used for adults. Sixty-eight patients were in the first quinquennium of life, 13 in the second, and 29 in the third. Eleven patients died of their tumor, three in the first quinquennium and eight in ght third. Our findings indicate that children who are less than 5 years old have a 7.3% chance of developing metastatic spread even though the local recurrence rate is 43%. Children who are 10 years old or older had a metastatic rate of 50% at 5-year follow-up, a figure closely approximating that for adults. We believe that infants and children less than 5 years old may be treated initially by local excision of the lesion, without sacrificing significant function of the part. Recurrent tumor does not seem to predispose to metastatic disease.


Induction of tumors in mice with the herbicide succinic acid 2,2-dimethylhydrazide.

A solution of 2% succinic acid 2,2-dimethylhydrazide was given continuously in the drinking water of 6-week-old randomly bred albino mice for the remainder of their lives. The treatment gave rise to tumors of blood vessels, lungs, and kidneys. The tumor incidences in these tissues in the controls were 6, 18, and 0%, whereas in the treated groups the corresponding tumor incidences were 73, 73, and 5%. Light microscopic examination revealed typical angiomas and angiosarcomas of blood vessels, adenomas and adenocarcinomas of lungs, and adenomas of kidneys. The study thus demonstrates the tumorigenicity of the herbicide, succinic acid 2,2-dimethylhydrazide. Since the residues of this chemical occur in fruit, the human population is exposed to it. The environmental implication of this finding and the fact that the hydrazines as a class have tumorigenic properties are discussed.


Growth stimulation of endothelial cells by simultaneous culture with Sarcoma 180 cells in diffusion chambers.

Endothelial cells derived from pig aorta were cultured in Petri dishes containing Sarcoma 180 cells growing in Millipore diffusion chambers. The presence of tumor cells consistently increased the colony-forming ability of the endothelial cells, an effect not produced by control-killed cells or 3T3 fibroblasts. This observation suggests that the cultured tumor cell retains the ability to produce diffusible substances that stimulate endothelial cell growth. These products may be similar to the angiogenic factors elaborated by malignant cells in vivo. The use of such a method may allow a closer study of the effect of tumor cells on the growth of a variety of tissues.


Transferrin promotion of 67Ga and 59Fe uptake by cultured mouse myeloma cells.

Radiotracer 67Ga-citrate is used as a tumor-seeking agent in clinical imaging investigations although fundamental reasons for its high uptake in certain malignant lesions remain unexplained. The mechanism by which 67Ga becomes concentrated in tumor cells has been investigated by comparing 67Ga and 59Fe uptake by cultured mouse myeloma cells with particular reference to uptake stimulation by transferrin. Concentrations of human transferrin down to 2 microgram/ml greatly stimulated cellular uptake of both tracers, whereas bovine transferrin proved relatively inactive. The rates of stimulated uptake of both tracers were similar as was their high degree of retention by cells, but their quantitative dependencies on transferrin concentration showed characteristic differences. Pretreatment of human transferrin with saturating amounts of nonradioactive Fe3+ canceled its ability to promote 59Fe uptake, but it had little effect on its promotion of 67Ga uptake. Further increase in the amount of added Fe3+ did cause a progressive depression of 67Ga uptake, but this effect probably relates to the iron distribution in the whole-cell culture system including the fetal calf serum component of cell growth medium. The results suggest that 67Ga and 59Fe reveal different aspects of the interaction of transferrin with cells.


Sensitivity of different cell lines and of different phases in the cell cycle to hyperthermia.

The sensitivity of different cell lines (Chinese hamster ovary, HeLa, L1210, and P388) to 43 degrees was compared Chinese hamster ovary and HeLa cells were much less temperature sensitive than were L1210 or P388 cells. This difference persisted even when HeLa and L1210 cells were grown in the same medium and suggested that this was an inherent difference between the cell lines. Of all the cell lines, 7-day L1210 ascites, maintained by transfer in mice, were the most sensitive. The greater sensitivity of these cells, as compared to 4-day L1210 ascites or cells in culture, may be explained by the difference in their growth stages. The 7-day ascites cells would be in stationary growth as compared to the exponentially growing 4-day ascites or the cells in culture. The temperature sensitivity of Chinese hamster ovary cells in different parts of the cell cycle was determined. Mid and late-S-phase cells were more sensitive than cells in mitosis or early S; G1 and G2 cells were the least sensitive. In a partially synchronized culture, the heat sensitivity of L1210 cells increased with increased percentage of cells in S phase.


Mantel-Haenszel analyses of litter-matched time-to-response data, with modifications for recovery of interlitter information.

The Mantel-Haenszel procedure for comparing sets of time-to-response data is adaptable to data that can be stratified on other variables. A particular adaptation, which we have used, is one in which animals from the same litter have been assigned to different treatment groups, e.g., some to a control group and some to a drug treatment group. The time to response used was that of tumor appearance; death from other causes was considered a loss to observation. The initial litter-adjusted analysis seemed to have only limited advantages compared to analysis that ignored litters and could be interpreted as suggesting that litter matchihg was not advantageous. Contributing to the difficulty was the fact that in the litter-matched analysis no further information was forthcoming from the remaining similarly treated animals in a litter when there were no remaining contrastingly treated littermates. Several devices for recovering interlitter information from such remnants and for combining it with intralitter information are examined and applied.


Retinoic acid-binding protein in experimental tumors and in tissues with metastatic tumor foci.

Screening for retinoic acid-binding protein (RABP) in experimental tumors revealed the presence of this protein in three mammary tumors, two metastatic colon tumors, B16 melanoma. Lewis lung carcinoma, Ridgway osteogenic sarcoma, and keratoacanthoma. RABP was below the limits of detection in two weakly metastatic colon tumors and in Sarcoma 180. After s.c. implantation of RABP-containing tumors into mice, this protein could be traced in the lungs due to pulmonary metastasis. Following implantation of Lewis lung tumors, RABP was detected in the lung on the 6th day. On the 15th day after implantation, RABP was present in lung and brain, but not in other tissues where this protein was normally lacking. In primary cultures of Lewis lung carcinoma, the lower limit for detection of RABP by sucrose gradient sedimentation technique corresponded to 0.12 mg protein that was extractable from 3 X 10(5) cells. Both chick embryo skin and rabbit ear skin extracts contained RABP; the level of cellular retinol-binding protein was high in chick embryo skin but only marginal in rabbit ear. The amounts of these proteins on chick embryo skin and rabbit ear skin correlate with the biological potency of retinol and retinoic acid, as observed by others.


Influence of histone H1 on chromatin structure.

Removal of histone H1 produces a transition in the structure of chromatin fibers as observed by electron microscopy. Chromatin containing all histone proteins appears as fibers with a diameter of about 250 A. The nucleosomes within these fibers are closely packed. If histone H1 is selectively removed with 50-100 mM NaCl in 50 mM sodium phosphate buffer (pH 7.0) in the presence of the ion-exchange resin AG 50 W - X2, chromatin appears as "beads-on-a-string" with the nucleosomes separated from each other by distances of about 150-200 A. If chromatin is treated in the presence of the resin with NaCl at concentrations of 650 mM or more, the structural organization of the chromatin is decreased, yielding fibers of irregular appearance.


A pseudogene structure in 5S DNA of Xenopus laevis.

The 5S DNA of Xenopus laevis, coding for oocyte-type 5S RNA, consists of many copies of a tandemly repeated unit of about 700 base pairs. Each unit contains a "pseudogene" in addition to the gene. The pseudogene has been partly sequenced and appears to be an almost perfect repeat of 101 residues of the gene. The order of components in the repeat unit is (5') long spacer--gene--linker--pseudogene (3') in the "+" strand (or H strand) of the DNA. The possible function of the pseudogene is discussed.


Recombinant DNA formation in a cell-free system from Xenopus laevis eggs.

A cell-free system is described which formed very high levels of recombinant DNA structures in 4 hr at 26 degrees C. It consisted of a single fraction of a high speed supernatant prepared from an extract of unfertilized eggs of the frog Xenopus laevis. This fraction eluted at 0.16-0.18 M Tris homogenization buffer from a DEAE-cellulose column. When two partially homologous supercoiled DNA molecules of different contour lengths were incubated simultaneously in this system, high levels of heterologous figure eight DNA structures were formed and observed by electron microscopy. Subsequently cleavage of the newly formed figure eight structures with Bam HI and Eco RI restriction endonucleases gave rise to "alpha structures" and "chi structures." The observed figure eight structures presumably represent the recombination intermediate predicted by the Holliday model for genetic recombination.


Translational control of protein synthesis during the early stages of differentiation of the slime mold Dictyostelium discoideum.

As analyzed by two-dimensional polyacrylamide gel electrophoresis, no new proteins are synthesized during the first 60 min of differentiation of Dictyostelium discoideum. The major change observed is the cessation of synthesis of five polypeptides and the reduction in the relative rates of synthesis of several more. We show here that this specific inhibition of protein synthesis is under translational control; the mRNAs for these proteins persevere in the cell in a translatable form for as long as 4 hr of differentiation, but these proteins are not synthesized by the cells after 2 min of development. As determined by analysis of the subcellular distribution of ribosomes and messenger RNA, there is a precipitous drop in the overall rate of polypeptide chain initiation during the first 5 min of differentiation. To interrelate and explain these phenomena, we show that a recent kinetic analysis of mRNA translation can explain how a reduction in the activity of a component of the initiation machinery required for translation of all mRNAs, such as an initiation factor, could result in a reduction in the overall rate of chain initiation and also a preferential inhibition of translation of certain mRNAs.


Animal dander allergy.

In a group of twenty-three atopic patients skin tests were performed with the dander allergens of horse, cat and guinea-pig, and with house dust and Timothy pollen allergens. A good agreement was observed between positive skin reactions and the results of RAST with these various allergens. In a number of cases, positive skin reactions were not related to the clinical histories. Likewise, positive RAST scores in several instances proved clinically meaningless. In patients with history-related skin reactions better correlation with RAST was observed. In such patients, skin reactions with photo-inactivated allergens remained strongly positive.


Pulmonary hypersensitivity to Alternaria and Aspergillus in baker's asthma.

In two cases of baker's asthma pulmonary hypersensitivity was found to the fungi Alternaria and Aspergillus. Provocative bronchial challenge revealed a dual response; an immediate and an Arthus type hypersensitivity to Aspergillus in the first case. A primary binding assay revealed high titres of anti-Aspergillus antibody in the serum. In the second case intradermal and bronchial challenge suggested an immediate type I hypersensitivity response to Alternaria. The suspected organisms were present in the room air of the bakeries. It is suggested that an immunological response to these airborne fungi may have contributed to the pathogenesis of baker's asthma.


A semi-automated fluorometric method for total estrogens in pregnancy urine.

We report a continuous-flow fluorometric method for total urinary estrogens that involves the Kober reaction, with extraction of the reaction product into dichloroethane containing trichloroacetic acid as described by Hahnel and Jones [Clin. Chim. Acta 16, 185 (1967)]. The dichloroethane extraction gives greater stability to the Kober color and sharper separation of aqueous and organic phases. The analytical system is adjusted to give maximum response to estriol 16alpha,beta-D-glucuronide, the principal estrogen conjugate in urine from late pregnancy, when calibrated with estriol standards. An initial 20-fold dilution of the sample with water increased analytical recovery of estriol conjugates from urine while maintaining adequate fluorescent response. Glucose interference was reduced by dilution and eliminated by treatment with sodium borohydride. Urinary protein up to 20 g/liter did not interfere. Total estriol, as determined by gas-liquid chromatography, comprises about 70% of total urinary estrogens in late pregnancy as measured by our continuous-flow fluorometric method. A reference range is presented based on 209 randomly collected urine specimens in which total urinary estrogens are expressed as a ratio to creatinine.


The foetus in Duchenne muscular dystrophy: muscle growth in tissue culture.

In tissue culture of muscle from five normal foetuses and four male foetuses at risk for Duchenne muscular dystrophy, no differences were found with regard to the ease with which cultures could be established and maintained, or in gross morphology or rate of growth and differentiation in culture. Yet before culture, muscle histology was clearly abnormal in at least one of the at-risk foetuses. The identification of the particular conditions of culture which prevent the full expression of the dystrophic process might be profitably pursued as a possible first step to finding a treatment.


Study on the binding of dihydrotestosterone, testosterone and oestradiol with sex hormone binding globulin.

Saturation studies on sex hormone binding globulin (SHBG) were performed with increasing levels of the three steroids: dihydrotestosterone (DHT), testosterone (T) and oestradiol (E2) in normal female sera. The curves obtained showed the clearly increasing SHBG affinity for E2, T and DHT, respectively. With T as saturating ligand, the mean values of SHBG binding capacity in normal and pathological sera, expressed as 10(-8) M SHBG were, for normal women, 5.87 +/- 0.2, for normal men, 3.69 +/- 0.1 and for hirsute women, 5.05 +/- 0.2. In hirsutism, SHBG levels were measured together with T and DHT levels. Hyperandrogenia was often better reflected by the T/SHBG ratio than by T levels alone. SHBG binding capacity, measured in pregnancy from the 7th to the 40th week, increased progressively as far as the 30th week. Values up to 5 times those of the mean value of the cycle were observed at the end of pregnancy. Variations in DHT, T and E2 binding as functions of SHBG concentrations, were also studied by adding a constant amount of each steroid to increasing SHBG concentrations in serum. DHT showed the greatest binding capacity, followed by T and then E2. Since the unbound fractions of T and DHT are believed to be biologically active, and since T and DHT are more sensitive than E2 to SHBG variations, the results obtained further support the hypothesis that SHBG plays an important role in the sex steroids balance.


Acute abdominal pain.

Acute abdominal pain is a daily problem in a busy pediatric practice and in hospital emergency rooms. An initial, carefully performed history and physical examination should differentiate the child who has a surgical disease from one who requires medical therapy or only reassurance. Whenever there is doubt about the diagnosis, the child must be admitted to the hospital for observation and appropriate laboratory examinations. This will allow for the early diagnosis of appendicitis and other diseases requiring an operation and will prevent needless operations on children with medical illnesses.


Meiotic studies of translocations causing male sterility in the mouse. I. Autosomal reciprocal translocations.

A new meiotic phenomenon is described in male heterozygous for the male-sterile translocations T(10;13)199H, T(16;17)43H, and T(7;19)145H. The phenomenon consists of a nonrandom contact between the C bands of the X chromosome and the translocation configuration in diakinesis/metaphase I plates. Translocation configurations with positively heteropycnotic regions, oftern associated with the allocyclic X chromosome, are found in some early diakineses that have not been overtreated with alkali. Such heteropycnosis of a part of translocated autosome, apparently in phase with the allocyclic X, is typical for all three male-sterile translocations. In contrast to these findings, neither nonrandom contacts nor positive heteropycnosis of the translocation configuration can be found in males heterozygous for the translocation T(9;17)138Ca, which does not impair spermatogenesis. Dissociation of the X and Y at diakinesis is significantly enhanced in sterile males, though the occurrence of dissociation is evidently not related to the presence of the C-band contact between translocated chromosomes and the X. A working hypothesis is proposed, relating the observed nonrandom C-band contact and heteropycnosis of translocated chromosomes to a presumed impairment of X inactivation in primary spermatocytes and to consequent failure of spermatogenesis. An alternative explanation cannot be excluded, however, which would account for the hitherto available data wihtout postulating any causal relationship between the meiotic findings and male sterility. Both alternatives are amenable to experimental verification.


Nonpairing of the X and Y chromosomes in the spermtocytes of BDF1 mice.

In the hybrid mouse strain BDF1, some 35--40% of spermatocytes had unpaired X and Y choromosomes in stages ranging from diplotene to first meiotic metaphase. This phenomenon varied significantly from mouse to mouse. In pooled material from Swiss Albino and CF1 mice, the corresponding frequency was 5.7%. In C57 BL/6 mice, one of the parent strains of BDF1 mice, the X and Y were separate in 7.7% of the spermatocytes. Based on the behavior of the X and Y in the BDF1 strain, it is concluded that they do not pair, rather than initially pairing and then precociously separating. The factor interfering with the pairing of the X and Y does not affect the autosomes; possibly it is an incompatibility of the two sex chromosomes, which come from different inbred lines.


The distribution of sequences complementary to human satellite DNAs I, II and IV in the chromosomes of chimpanzee (Pan troglodytes), gorilla (Gorilla gorilla) and orang utan (Pongo pygmaeus).

Human satellite DNAs I, II and IV were transcribed to yield radioactive complementary RNAs (cRNAs). These cRNAs were hybridised to metaphase chromosomes of man, chimpanzee (Pan troglodytes), gorilla (Gorilla gorilla) and orang utan (Pongo pygmaeus). The results of this in situ hybridisation were analysed quantitatively and compared with accepted chromosome homologies based on Giemsa banding patterns. The cRNA to satellite II (cRNAII) did not hybridise to chimpanzee chromosomes, although its hybridisation to chromosomes of gorilla and orang utan yielded more autoradiograph grains than hybridisation to human chromosomes, and cRNAIV hybridised to many chromosomes of gorilla and chimpanzee but was almost entirely restricted to the Y chromosome in orang utan. Most sites of hybridisation were located on homologous chromosomes in all four species, but there were a number of sites which showed no correspondence between satellite DNA location and chromosome banding patterns, and others where a given chromosomal location hybridised with different cRNAs in each species. These results are in contrast to those found for many transcribed DNA sequences, where the same sequence is usually located at homologous chromosome sites in different species, and appear to cast doubt on many proposed models of satellite DNA function.


The case of the numb testicles.

Both psychiatrists and neurologists must be able to recognize illnesses that have atypical presentations so as to insure prompt and accurate diagnosis and treatment. The disease described in this case report graphically illustrates this point.


FSH and LH stimulation of ornithine decarboxylase activity: studies with porcine granulosa cells in vitro.

Under defined conditions in vitro FSH and LH caused a dose-dependent stimulation of ornithine decarboxylase (ODC) activity in granulosa cells isolated from porcine ovarian follicles. In cells from small follicles, FSH was at least twice as potent an inducer of the enzyme activity as LH. The cells from medium-sized follicles were more responsive to both hormones than cells from small follicles. In addition, the cells from medium-sized follicles were approximately equally responsive to maximal FSH and LH stimulation. Incubation of cells from either small follicles or medium-sized follicles with maximum effective doses of LH plus FSH caused no additive effects.


Various effects of arsenic in Japan depending on type of exposure.

Since 1955, a number of outbreaks of arsenic poisoning have occurred throughout Japan among industrial workers and the general population. The sources and types of exposure differ according to the incident, and the victims vary in sex and age. These incidents include arsenic poisoning in milk, soy sauce, and well water, pollution originating from the Toroku Mine on the island of Kyushu, the Matsuo Mine in Shimane Prefecture and Saganoseki Smelter on Kyushu. The type, routes, and dose of exposure as well as major symptoms are given. The clinical signs and symptoms are discussed in relation to the various types of exposure which occurred in these incidents. Dose-response relationships will be considered where data are available.


X-ray investigation of the binding of 1,10-phenanthroline and imidazole to horse-liver alcohol dehydrogenase.

We have studied the binding of two inhibitor molecules, imidazole and 1,10-phenanthroline, to liver alcohol dehydrogenase by crystallographic methods. X-ray data for the imidazole complex were collected to 0.29-nm resolution and for the 1,10-phenanthroline complex to 0.45-nm resolution. In both cases we found only one peak in the difference electron density maps close to the active zinc atom. The peak corresponding to 1,10-phenanthroline overlaps the site of the density of the zinc-bound water in the apoenzyme and the imidazole density partly overlaps this density. We can not discern any additional peaks close to the zinc atom which would correspond to new positions of bound water. We thus conclude that both these inhibitors bind to the catalytic zinc atom and that upon binding they displace the water molecule that is firmly bound to this zinc atom in the apoenzyme. We do not see any structural changes in the remaining part of the molecule.


Demonstration of angiogenesis-activity in the corpus luteum of cattle.

The angiogenetic potency of corpus luteum tissue and of extracts of it was tested in three different biological model systems: the chorioallantoic membrane of the chicken (CMA), the ventral subcutaneous pouch of the mouse and the cheeck pouch of the Syrian hamster. The formation of new capillaries after transplantation of the materials was scored macroscopically and stereomicroscopically. In all three systems strong capillary reactions at the periphery of the transplants could be observed indicating the presence of a capillary inducing potency in the tissue of corpus luteum of the cattle.


The effect of a low protein diet on the adipocytes of rat perigenital fat.

Weanling rats were fed a low protein diet for 6 weeks and their weights were 50% less than controls. There were significantly fewer adipocytes per microgram adipose tissue, but estimates of the number of adipocytes per rat indicated that the diet had much less effect on adipocyte number than on b. wt.


Induction of rotational behaviour by intranigral baclofen suggests possible GABA-agonist activity.

In rats, unilateral injections of the GABA-derivative baclofen into the zona reticulata of the substantia nigra produced a contralateral rotation that was translated to ipsilateral rotation under the influence of amphetamine. These results mimic those following unilateral elevation of GABA levels in the substantia nigra and suggest that baclofen may have some GABA agonist activity following intracerebral injection.


The effects of intramuscular injections of 15(S)-15-methyl prostaglandin F2alpha in failed abortions.

Intramuscular injections of 15(S)-15-methyl prostaglandin F2alpha (15-Me-PGF2alpha) induced abortion in 38 patients who had failed to abort with other techniques, such as intra-amniotic instillation of saline or PGF2alpha and intravaginal insertion of prostaglandin-impragnated Silastic devices. The intramuscular injections of 15-Me-PGF2alpha were initiated when the original abortion techniques, even when augmented by intravenous oxytocin, failed to produce expulsion of the fetus. The dose schedule was 250 microgram or 500 microgram every 2 to 4 hours, and the concomitant intravenous oxytocin was continued at a rate of 167 mU/minute. Of the 38 patients, 26 aborted with two or fewer injections of 15-Me-PGF2alpha, and 30 patients required only 1 mg of the drug to expel the fetus successfully. The mean time from the first injection of 15-Me-PGF2alpha to the expulsion of the fetus was 5.25 hours; one-half of the patients aborted in less than 4 hours. The placenta was expelled spontaneously in 15 patients, removed manually from the vagina in 18, and removed by sponge forceps in 3. Two abortions were incomplete and surgical intervention was required. Twenty-eight patients (74%) experienced gastrointestinal disturbances, chiefly vomiting and diarrhea. Intramuscular administration of 15-Me-PGF2alpha eliminates the need for repeated amniocentesis, and the dose may be adjusted to meet the precise requirements of the clinical situation.


Early first-trimester abortion induction by Silastic vaginal devices for continuous release of 15(S)=15-methyl prostaglandin F2alpha methyl ester.

The abortifacient efficacy of four Silastic devices containing 15(S)-15-methyl prostaglandin F2alpha methyl ester (15-Me-PGF2alpha methyl ester) was evaluated by single vaginal insertion and retention for 24 hours in four groups of ten patients each at 31 to 49 days beyond the 1st day of the last menstrual period. There were eight complete and two incomplete abortions in the group receiving a device with a drug concentration of 0.5% and a surface area of 20 sq cm. Four complete abortions, four incomplete abortions, and two failures occurred with a 0.2%, 10 sq cm device. Seven complete and two incomplete abortions were obtained with a 0.5%, 10 dq cm device; one patient in this group was found to have aborted prior to treatment. A 0.5%, 2o sq cm device with one-half the thickness of the other 0.5% devices yielded five complete abortions, one incomplete abortion, and four failures. Side effects were minor and well tolerated in each group. Serial determinations of plasma progesterone, human chorionic gonadotropin, and 15-Me-PGF2alpha were obtained and related to the outcome of therapy. Because of failure to achieve sustained blood levels of drug or satisfactory rates of complete abortions, this technique, with the devices used in this study, is not believed to be acceptable for routine early first-trimester pregnancy termination.


Abortion of early pregnancy on an outpatient basis using Silastic 15(S)-15-methyl prostaglandin F2alpha vaginal devices.

Hormonal changes associated with the use of 15(S)-15-methyl prostaglandin F2alpha in a Silastic device as an abortifacient were evaluated during very early first-trimester pregnancy. Five of six patients aborted completely with drug alone. Measurement of plasma samples demonstrated rapid drug absorption with apparent effective concentrations of 600 to 3000 pg/ml but without a pattern of sustained release and absorption. Routine serial determination of both serum chorionic gonadotropin (hCG) and beta-subunit hCG were used to assess the efficacy of treatment. Although more sensitive, beta-hCG measurement offered no advantage clinically over the standard hCG assay. Plasma progesterone values also reflected success or failure of the technique. Vomiting and diarrhea continue to be problems associated with the use of these compounds. The Silastic device may be the forerunner of a self-administered early abortifacient. Continued efforts are necessary to elucidate mechanisms of action and to find an analog specific for myometrial stimulation.


[Effect of alcoholic intoxication on the activity of several dehydrogenases and aminotransferases in rat brain tissue].

An acute intraperitoneal administration of ethanol (4-8 g/kg) produced a fall in the activity of malate-dehydrogenase (MDG), alanine- and aspartate-aminotransferase in homogenates and mitochondria of the cerebellum, large hemispheres and liver of the rats by 25 and 50 per cent, respectively. The activity of succinate-dehydrogenase (SDG) increased by 24 per cent in the large hemispheres and fell by 20 per cent in the liver and in the cerebellum homogenate. The activity of lactate-dehydrogenase (LDG) of the large hemispheres and liver mitochondria decreased by 30 per cent. A chronic (1 1/2-2 month long) ethanol intoxication produced a drop of the LDG and MDG in the mitochondria of the nerve structures and of aminotransferase in the liver. The SDG activity rose in the homogenate of the cerebellum, large hemispheres and of the liver by 80, 50 and 20 percent, respectively. The dehydrogenases and aminotransferases activity in the liver and nerve structures of rats in the state of abstinence was somewhat below the normal, but higher than in animals chronically consuming alcohol.


[Pharmacokinetics of 35S-tomizin in mice with leukemia L-1210].

A comparative study of the distribution of 35S-tomisin in the organism of intact (without neoplasia) mice and those with L-1210 leucosis was carried out. In both animal groups the tagged drug was found to be largely accumulating early after its introduction in the tissues of the liver, kidneys, intestines, spleen and lymph nodes. In 30 minutes time following introduction of 35-S-tomison 7.3 +/- 0.7 per cent of its amount was detected in the ascitic fluid. On centrifugation of the tagged drug its bulk (92-95 per cent) remained in the supernatant fluid. During the first 24 hours the drug leaves the body: 25-35 per cent with urine, 3-5 per cent--with feces and 0.1-0.3 per cent--with bile. In the urine of mice with leucosis the products of the 35S-tomisin biotransformation appear earlier than in intact animals. The most marked immunodepressive effect of tomisin was marked with its administration 24 and 48 hours after immunization.


[Effect of single and chronic administration of ethanol on several metabolic indices in animals].

The effect of one-time and 10-day long administration of ethanol (2 ml of  a 20% solution per 100 g of body weight) on the activity of alcohol-, lactate-dehydrogenase, aspartate- and alanine-aminotransferase in cytostructures of different rats' organs was studied. One-time introduction of ethanol intensifies the processes of the amino acids transamination in individual cytostructures, reduces the activity of alcohol- and lactate-dehydrogenase. A course-wise administration of ethanol produces a still greater reduction of their activity.


Pregnant mare's serum gonadotropin. III. Hemispaying and the reversal of the antifertility faculty of pregnant mare's serum gonadotropin.

A single injection of 10 IU of pregnant mare's serum gonadotropin (PMSG) on day 5 of pregancy caused wastage of the fetoplacental unit by day 16 of pregnancy. Semispaying, which seems to subtract approximately 50% of the ovarian steroid contribution, at 24, 48, and 72 hours following the PMSG schedule prevented the antifertility faculty of the hormone preparation. However, hemicastration at 96 hours following the gonadotropin regimen was found to be ineffective, and 100% of the test animals showed complete termination of pregnancy. Experimental data collectively tempt us to propose that an estrogen excess, particularly of follicular origin in both ovaries, is essential for more than 72 hours following gonadotropin sensitization before the antifertility faculty of PMSG can be demonstrated.


Action of A RNase and AS RNase on growth of cells in vitro.

The effect of bovine pancreatic A RNase and bull seminal vesicle AS RNase on proliferation of HeLa cells and LEP cells in vitro was studied. The ribonucleases were used in doses of 0.1, 1.0, 10.0, and 100.0 microgram per ml of medium and 100 000 inoculated cells. Proliferation was evaluated by the growth curves. Both in single and long-term experiments. A RNase in all doses used had no effect on cell proliferation, whereas AS RNase exerted inhibitory activity beginning with the dose of 10 microgram, and affected more markedly the LEP cells (a diploid non-malignant cell line) than the malignant heteroploid HeLa cells. When added to the cell suspension, AS RNase was shown by indirect immunofluorescence to bind to HeLa cell membranes in all phases of growth, whereas AS RNase was found on membranes of LEP cells only when they were derived from the log phase of growth.


[Monitoring of the results of the treatment of chorionic carcinoma (author's transl)].

Because of the high sensitivity and specificity of the method, radio-immunologic determination of the Beta Subunit HCG in the serum is superior to other methods of monitoring the therapeutic results of the treatment of chorionic carcinoma. A ratio of 225 to 1 of HCG concentrations in the serum versus the cerebral spinal fluid is evidence against the probability of a metastasis of chorionic carcinoma in the central nervous system. Following successful treatment of chorionic carcinoma it is advisable to determine the serum HCG with the Beta Subunit method weekly during the first two months and montly in the succeeding two years and thereafter at three months intervals.


[Report of a case of trichophyton fungus granulomatosis of the breast and its differential diagnosis to inflammatory carcinoma (author's transl)].

The case of a 34 year old patient is reported who suffered from a chronic dermatatomycosis. The trichophyton fungus granula matosis secondarily invaded the parenchyma of the breast and clinically first presented with signs and mammographic findings suggestive of an inflammatory carcinoma. A similar case report was not located in the international literature. Therefore the clinical and mammographic details and the conclusive microscopic findings of the biopsy of this case are reported for the purpose  of differential diagnosis.


[Studies on the physical dependence liability of chlorphenesin carbamate (author's transl)].

Physical dependence liability of chlorphenesin carbamate (CPC) was studied in parallel with phenobarbital-Na (PB). Beagle dogs were used and the overall duration of the experiment was 85 days, i.e. the first dosing period was 42 dyas (6 weeks) in which drugs were repeatedly administered orally once daily, followed by a withdrawal period (7 days), the second dosing period was continued from the 50th-78th day in which the form and schedule of drug administration was as in the first dosing period. The last 79th to 85th days were used for substitution experiments. In both dosing periods, PB but not CPC showed signs of tolerance formation. Severe withdrawal syndrome was observed in PB administered dogs whereas there were no changes of behavior observed in CPC-dogs by withdrawal and substitution procedures, respectively. CPC apparently does not have a physical dependence liability.


Mechanism of histopathological changes of nerve cells experimentally induced by chronic alcohol poisoning.

Experimental alcoholism was produced in rats by supplying them with 15% ethanol as the only source of liquid for a whole year. Histopathological examination revealed that Purkinje cells and granule cells in the cerebellum mainly showed such changes as decrease of ER, ribosomes and severe atrophy of the nerve cells. It might be speculated that these changes were caused by the disturbance of protein synthesis in the nerve cells induced by chronic alcohol effect.


Radioimmunoassay of melatonin: human serum and cerebrospinal fluid.

Using a newly characterized anti-melatonin serum it has been possible to establish human serum melatonin concentrations at short time intervals during 24h. A clear circadian rhythm with peak values during the dark phase was demonstrated in both men and women. Values (pg/ml;mean +/- SE) were as follows: females 02.00h: 130 +/- 7, 18.00h: 45 +/- 5 males 02.00h: 140  +/- 11, 18,00H: 70 +/- 5. The estimation (pg/ml; mean +/- SE) of melatonin in human serum (males: 63 +/- 22 and females: 100 +/- 45) and cerebrospinal fluid simultaneously taken has shown that melatonin is lower in cerebrospinal fluid (males: 59 +/- 33 and females: 57 +/- 28). Blanks are not subtracted.


Isochromosome Yq in a woman with atypical Turner's syndrome.

A female with 46,X,i(Yq) in all cells and a survey of previous cases of isochromosome Yq is presented. She was first admitted to hospital 15 years old due to nanismus and retarded sexual development. Gonadal dysgenesia was observed, and the diagnosis 'atypical Turner's syndrome' was applied. The patient, who presents only a few Turner stigmata, has been given cyclic estrogen treatment since the age of 16. She has developed normal secondary sex characteristics, cyclic bleedings and has attained normal height (161 cm). Since the age of 18 the patient has suffered various periods of anemia caused by gastrointestinal hemorrhage. This hemorrhage is probably due to intestinal teleangiectasiae which are found with increased frequency in patients with Turner's syndrome.


Estimating the parameters of the incompletely penetrant single locus model using multiple populations.

A method involving the comparison of two or more populations is suggested as a means of obtaining a unique solution to the parameters of the incompletely penetrant single locus model. The proposed method allows a test of the assumptions of the model when three or more populations are compared. Equations that allow the inclusion of data on twin concordance rates and/or the proportion of affected children given neither, one or both parents affected are also given. Finally, some implications of fitting the model are discussed in terms of genetic counseling, residual environmental variance and the concept of heritability as applied to dichotomous traits.


Palmar dermatoglyphic patterns in twins.

The role of genetic factors in the determination of palmar dermatoglyphic patterns was investigated in a series of 49 MZ and 51 DZ twins, using Spearman's rank correlation and analysis of variance. Both methods indicated that the genetic effect in the distribution of patterns is highest in the interdigital III and lowest in the interdigital IV regions, the hypothenar and thenar showing intermediate values. As for interdigital II, no evaluation of genetic effects was possible using the nonparametric test, but the estimates of genetic variance indicate that inherited factors may play a relatively minor role in the pattern distribution of this area.


Long-term organ culture of mouse mammary gland.

A method for maintaining mouse mammary gland in organ culture for periods of at least 30 days is described. Strips of the number four mammary glands were cultured in individual tubes while fully submerged in Medium 199 supplemented with insulin, aldosterone, ovine prolactin and bovine growth hormone. Exchange processes were aided by slowly rotating the tubes during culture. Mammary tissue from midpregnant BALB/c and virgin GR/A mice was induced to undergo lobulo-alveolar development, secrete and remain differentiated and metabolically active for the period of culture. Cells of both the ductal and alveolar epithelium continued to synthesize DNA and divide. The submerged roller-tube culture allows the use of larger pieces of tissue than can be accommodated in static culture, and the technique may prove applicable to culture of a variety of tissues.


Intracellular levels of adenosine triphosphate in hamster trachea organ cultures exposed to Mycoplasma pneumoniae cells or membranes.

The amount of adenosine triphosphate (ATP) in hamster trachea organ cultures was determined with a technique based on light emission from a luciferin/luciferase/ATP reaction. The amount of ATP, expressed as ng per mg dry weight, was consistent in tracheal explants prepared from various animals and changed negligibly when explants were cultivated in vitro for several days. The amount of ATP was related directly to cellular activity and integrity in the epithelium since inactivation by heat or freeze-thaw rapidly depleted measurable ATP, and ciliary activity and ATP content were related directly. When tracheal explants were infected with 10(5) to 10(7) CFU of virulent Mycoplasma pneumoniae cells, both ciliary activity and ATP content in the tissue dropped dramatically after approximately 5 to 8 days (up to 85% and 60% decreases, respectively). Exposure of explants to 50 to 200 microgram per ml of purified M. pneumoniae membranes also caused significant decreases in ciliary activity and ATP. When explants were infected with attenuated or nonvirulent mycoplasmas, ciliary activity was only slightly decreased, while ATP values often rose slightly. The technology associated with the determination of ATP levels in tracheal explants should prove useful as a new, objective, analytical approach to cell viability in organ cultures.


An improved organ culture method for adult mammalian lung.

An improved method for maintaining adult rat lung in submerged organ culture is described in which the alveoli were inflated with agar and 200-micron-thick hand-cut sections were mounted in Rose chambers. The conventional single-compartmented Rose culture chamber was modified by adding a second chamber separated from the first by a gaspermeable membrane. One compartment functioned as an air reservoir and the other housed the explants submerged in nutrient medium. Visking dialysis membrane used underneath the explants prevented cell outgrowth and facilitated the exchange of nutrients and waste products at the glass-tissue interface. Because of the excellent optical properties of the Rose chamber and the thinness of the explants, individual cell types can be identified in the living tissue. The explants were studied with time-lapse cinematography, light microcoscopy, histology, and erythrosine B for dye exclusion. With this modified system the functional life span of the explants was increased from 1 week to 1 month.


[Preventive-medicine screening methods in the examination of blood donors].

From a blood transfusion service with ca. 7000 donors a year, there was a loss of donors whose blood, having undergone medical tests, was found to be unsuitable. Details of all such donors were carefully documented. The results were ascertained by attentive control of the health of the donors which, in part, exceeded the minimum requirements laid down by the existing regulations. The relatively high loss of donors and the kind of disease underline the importance of these control checks, as an aspect of preventive medicine. This is of particular importance with regard to the results of so-called "new donors", who register for the first time. They had to undergo examination, and were only allowed to give blood when all the results of the tests had been submitted. The question then arises as to whether the regulations for the differing minimum requirements for the examination of "occasional donors" and "regular donors" can be maintained. A reduction of the expenditure on the present regulation examination of donors is not advocated because of both responsibility towards the blood donors, and in view of the increasing significance, to the medical care of patients, of the proximity of a clinic to an expedient transfusion service.


[Perioperative infusion therapy during childhood. II. Balance studies on pre-, intra- and postoperative infusion of basic solutions].

For a period of 63 hours totally, 4 different infusion solutions were administered to infants and children of 3 age groups (I = 10th day to the 6th month of age, II = 6th month to the 3rd year of age and III = above 3 years of age) during the pre-, intra- and postoperative period. The serum sodium, potassium-, chloride-, phosphate- and blood urea concentrations remained completely unchanged as well as the acid base parameters. Even between the age groups, no significant differences could be observed, as far as the above mentioned criteria are concerned. But the blood glucose concentrations increased within all age group in the immediate postoperative phase. The potassium concentrations in the urine decreased throughout in age group I. From these results, one can conclude, that these specific infusion solutions for paediatric surgical procedures with exception of solution I contained the most important electrolytes adaequately and that the amounts of fluids, which were given can be considered as sufficient. Nevertheless, the question remains, whether these different types of infusion therapy can be simplified by using more standardized solutions. This problem was subject to more sophisticated studies, which will be reported in the 3rd part of this publication, to be published at a later date.


[Faulty nutrition as viewed by the hepatologist].

Looking back over the past centuries, a survey shows that for the past 2000 years definite dietetic rules have existed for the treatment of liver diseases, which to a great extent have disregarded the distribution of protein and carbohydrates. Deviations from the principles of nutrition followed as a result of intensive physiological-chemical research during the 19th century. This new knowledge, however, lead to misinterpretation and exaggerations in the recommended dietary proportions of fat, protein and carbohydrates. Protein deficiency, which is particularly disadvantageous in cases of hepatitis during pregnancy, and lack of vitamins, along with other types of malnutrition, became apparent. Various attempts providing diets rich in protein and fat, for the treatment of hepatitis, proved rather the tolerance of these nutrients than discovering a new, more efficient therapy. In spite of some resignation, recommendations made on the basis of experience should rather be dispensed with, as regards diet, and the search for further knowledge of the supply of nutrients in liver diseases should be continued. In addition, the study of endotoxins and toxic amines in the metabolism of the brain offers a source of future research.


Mitogenic stimulation of murine spleen cells by brief exposure to Staphylococcus aureus enteroxin B.

Staphylococcal enterotoxin B rapidly and firmly attached to external membrane surfaces of C57BL/6 spleen cells and, unless neutralized by specific antitoxin, committed the spleen cells to strong blastogenic responses.


An abnormal luteal-phase evening peak of plasma prolactin in women with a family history of breast cancer.

In earlier study an abnormally high lutealphase evening peak of plasma prolactin in women with a family history of breast cancer was first noted, but there were few observations. The current study confirms the original findings. Other groups, e.g. nulliparous women and tall and heavy women also appear to show the same abnormality. Further work is required to establish this finding. However, the abnormally high luteal-phase evening peak of plasma prolactin in the family history group is still present when such women are disregarded. The possible relevance of the restriction of elevated prolactin levels, in enhanced-risk women, to the luteal phase, when both oestradiol and progresterone levels are also elevated, is discussed briefly as is the question why a minor peak at 19.00 h is important when plasma prolactin levels show a massive increase between 03.00 and 09.00 h.


Secondary amyloidosis and the serum amyloid precursor in leprosy: geographical variation and association with leukocytosis.

The prevalence of the amyloid-related serum component, protein SAA, was investigated in two groups of leprosy patients from different areas of Papua New Guinea. Protein SAA was more prevalent in coastal leprosy patients (49% positive) than in highland patients (21% positive). Paradoxically, many more cases of amyloidosis were diagnosed in the highland group (17 of 199) than in the coastal group (3 of 112). In the highland patient group, SAA was found to correlate with the leprosy disease spectrum, being more prevalent in patients toward the lepromatous pole. Borderline and tuberculoid patients who had detectable SAA usually had neurotrophic ulcers. No such relationships were observed in the coastal patient group, probably because other infections, more common on the coast, were also responsible for causing increased concentrations of SAA which is known to behave as an acute phase reactant. A correlation was observed between SAA positivity and neutrophil leukocytosis. This suggests that various inflammatory stimuli such as erythema nodosum leprosum reactions, neurotrophic ulcers and intercurrent infections, all contribute to the prevalence of SAA in leprosy patients.


Effect of urethral stretch on urethral pressure profile.

Urethral pressure profiles were recorded in anesthetized dogs with the urethra in both the relaxed and stretched positions. Similar studies were done with fresh specimens of intact dog bladder and urethra removed en bloc. Clinical studies were performed on women during surgical correction of stress incontinence. The pressure profiles reflected the intrinsic smooth muscular activity of the urethra, inasmuch as they were all obtained under full relaxation by anesthesia and with the abdomen open. In all cases, functional as well as anatomical lengths of the urethra were increased with stretch. The rise in urethral closure pressure was most marked in the proximal third of the urethra. Changes were more notable in women with stress incontinence than in the normal dog. Similar results were noted in the in vitro studies. Length-tension properties of smooth and striated muscles offer a clear explanation for the rationale and effectiveness of surgical procedures done for the correction of stress incontinence.


Effect of pelvic and retroperitoneal surgery on the urethral pressure profile and perineal floor electromyogram in dogs.

Study of the urethral pressure profile and perineal floor electromyogram in 12 dogs subjected to colonic resection, abdominal vascular stripping, and pudenal nerve transection suggest that specific defects in urethral continence function may follow pelvic and retroperitoneal surgical dissection. These alterations, attributable to damage of the pudendal nerves and the sympathetic fibers in the posterior peritoneum, must be accurately evaluated before treatment of postoperative urodynamic problems.


Acute renal failure in leprosy.

Three patients having lepromatous leprosy developed acute renal failure. Two patients completely recovered and one was left with a moderate degree of renal insufficiency. Renal tissue obtained by percutaneous biopsy revealed acute tubular necrosis in two and diffuse crescentic glomerulonephritis in the third case.


Striated muscle determinants of intraurethral resistance.

Urethral resistance was measured in dogs subjected to alterations in periurethral striated muscle activity before and after muscle paralysis. Changes in periurethral striated muscle activity that could be abolished by curare significantly affected intraurethral resistance measurements. Control of striated muscle is necessary for true assessment of other urinary sphincter components.


Atresia coli in a twin calf.

Atresia coli was diagnosed in a newborn calf. The calf was an identical twin, as evidenced by identical blood type and identical electrophoretic patterns for hemoglobin, transferrin, and amylase. The other calf was clinically normal. The calves resulted from embryo transfer and represented cleavage of a single embryo in the recipient animal. These findings indicate atresia coli in this calf was probably not hereditary.


A histopathologic study of renal biopsies in fifty cases of leprosy.

Renal biopsies from 50 cases of leprosy, including 45 cases of lepromatous and 5 cases of tuberculoid, have been studied in detail histopathologically with special reference to any specific leprosy lesion such as the presence of leproma or granuloma, the presence of acid-fast bacilli and the occurrence of amyloid deposit. Leproma or granuloma, acid-fast bacilli and amyloid deposit could not be detected in any of these cases. Pathologic features of nephritis of various types were seen in only 40% of cases. Similar observations made by previous authors have been reviewed. The question is raised as to why kidney tissue should escape from developing specific leprosy lesions in either type of leprosy when other tissues such as liver, striated muscles and lymph nodes are known to develop such lesions. A greater immunologic resistance of the renal tissue to lepra bacilli or local physiochemical factors which may render renal tissue an unfavorable site for the settling and multiplication of lepra bacilli are considered as possible related factors.


Structure of the tubulin dimer.

Microtubules are formed from a 110,000-dalton dimeric subunit called tubulin. Two forms of 55,000-dalton monomer, alpha and beta, are found in all microtubule preparations. The dimers could thus theoretically be either heterodimers (alphabeta) or homodimers (alphaalpha and betabeta). This problem was investigated by stigated by chemical cross-linking using several bifunctional reagents, of which one, dimethyl-3,3-(tetrame thylenedioxy) dipropionimidate dihydrochloride (DTDI), was able to make intradimer bonds in tubulin. When soluble chick brain tubulin was cross-linked with DTDI and analyzed by electrophoresis in an acrylamide gel system capable of resolving alphaalpha, alphabeta, and betabeta, 60 to 90% of the cross-linked dimer was alphabeta. If tubulin was incubated at 24 degrees prior to cross-linking with DTDI the total yield of cross-linked dimer increased with time, indicating that tubulin was forming loose aggregates. The relative amounts of cross-linked dimer alphaalpha and betabeta also increase with time, indicating that soluble tubulin is largely alphabeta, and suggesting that cross-linked alphaalpha and betabeta arise from nonspecific aggregation during tubulin purification. The aggregation observed by cross-linking with DTDI was strongly influenced by colchicine and Vinca alkaloids in a pattern similar to the effects of these drugs on tubulin polymerization.


The amino acid sequence of Clostridium pasteurianum iron protein, a component of nitrogenase. I. Tryptic peptides.

A total of 25 tryptic peptides was isolated from the S-beta-carboxymethyl derivative of Clostridium pasteurianum iron protein (N2). In order to obtain the various peptides in pure state, a combination of gel permeation, cation and anion exchange column chromatographic methods, as well as various ascending paper chromatographic methods were adopted. Sequence studies of the tryptic peptides were carried out mainly by a modified manual Edman degradation procedure and also by automated analysis, carboxypeptidase digestion, and by hydrazinolysis. Thus, 242 residues (88.6%) out of a total of 273 amino acid residues were sequenced in the present study. The sum of the amino acid residues in the tryptic peptides isolated from iron protein (N2) accounted for the 273 amino acid residues present in the iron protein.


The amino acid sequence of Clostridium pasteurianum iron protein, a component of nitrogenase. II. Cyanogen bromide peptides.

A total of 10 cyanogen bromide peptides were isolated from the S-beta-carboxymethyl iron protein of nitrogenase. Purification of these peptides was performed mainly by gel filtration on Sephadex G-50; by ascending paper chromatography using the solvent system of pyridine, isoamyl alcohol, 0.1 M ammonium hydroxide; and also, in some cases, with additional steps such as anion exchange column chromatography on Dowex 1-X2 or ascending paper chromatography in an acidic solvent system or by pyridine precipitation of the cyanogen bromide fragment. Sequenator analyses of three large cyanogen bromide peptides (53 to 72 residues) provided tryptic peptide overlap data for the inner portion of the protein. The cyanogen bromide peptides accounted for all of the 273 amino acid residues which were present in the tryptic peptides isolated from carboxymethyl-iron protein (Tanaka, M., Haniu, M., Yasunobu, K. T., and Mortenson, L. E. (1977) J. Biol. Chem. 252, 7081-7088).


Decarboxylation-dependent transamination catalyzed by mammalian 3,4-dihydroxyphenylalanine decarboxylase.

In addition to the usual decarboxylation, pig kidney 3,4-dihydroxyphenylalanine (dopa) decarboxylase catalyzes a decarboxylation-dependent transamination which converts dopa into 3,4-dihydroxyphenylacetaldehyde and sinultaneously converts enzyme-bound pyridoxal-P into pyridoxamine-P. Similar reactions occur when this enzyme acts on m-tyrosine, alpha-methyldopa, and alpha-methyl-m-tyrosine. The transamination occurs in about 0.02% of decarboxylations of dopa and m-tyrosine and in about 2% of decarboxylations of alpha-methyldopa and alpha-methyl-m-tyrosine. The fraction of decarboxylations proceeding by the transamination pathway is independent of pH. This reaction appears to result from a divergence in the normal mechanism of decarboxylation; the quinoid intermediate which is formed by decarboxylation of the substrate-pyridoxal-P-Schiff base ordinarily protonates on the alpha carbon of the amino acid, but protonation occasionally occurs at the benzylic carbon of the coenzyme, and this latter route leads to transamination.


The compressive behavior of bone as a two-phase porous structure.

Compression tests of human and bovine trabecular bone specimens with and without marrow in situ were conducted at strain rates of from 0.001 to 10.0 per second. A porous platen above the specimens allowed the escape of marrow during testing. The presence of marrow increased the strength, modulus, and energy absorption of specimens only at the highest strain rate of 10.0 per second. This enhancement of material properties at the highest strain rate was due primarily to the restricted viscous flow of marrow through the platen rather than the flow through the pores of the trabecular bone. In specimens without marrow, the strength was proportional to the square of the apparent density and the modulus was proportional to the cube of the apparent density. Both strength and modulus were approximately proportional to the strain rate raised to the 0.06 power. These power relationships, which were shown to hold for all bone in the skeleton, allow meaningful predictions of bone tissue strength and stiffness based on in vivo density measurements.


Heat-induced reversible hexagonal packing of spindle microtubules.

Epithelial cells cultured from the lung of the Northwest rough-skinned newt (Taricha granulosa granulosa) were subjected to brief (10-15 min) elevated temperature shocks of 33 degrees-36 degrees C during metaphase. Electron microscope studies on these cells reveal that the spindle microtubules (Mts) are differentially stable to heat treatment. The great majority of nonkinetochore Mts are destroyed within the first few minutes of the shock while kinetochore and adjacent Mts rearrange to form hexagonal closely packed structures before disassembling, the latter occurring only after prolonged heat treatment. The significance and theoretical implications of the formation of hexagonal closely packed Mt structures and of the differential stability of spindle Mts to heating are discussed. The data suggest the existence of one or more heat-sensitive structural component(s) which maintain the individual minimum spacing seen between spindle Mts. To our knowledge, this is the first reported instance of the experimental rearrangement of kinetochore Mts into reversible, hexagonal closely packed bundles.


Homogeneous cell populations from rabbit kidney cortex. Proximal, distal tubule, and renin-active cell isolated by free-flow electrophoresis.

A single-cell suspension has been prepared from rabbit kidney cortex by using a Ca-binding medium and gentle mechanical forces. The suspension was subjected to carrier-free electrophoresis, and several cell fractions were obtained. Proximal and distal tubule cell populations could be identified by their morphology. Renin-containing cells were located by means of radioimmunoassay. The morphology of the cells and their vitality (uridine incorporation) are discussed.


Membrane transport by guinea pig peritoneal exudate leukocytes: effect of phagocytosis on hexose and amino acid transport.

Short term, carrier mediated transport of D-glucose, L-leucine and L-lysine by guinea pig peritoneal macrophages was characterized. Analysis of the amino acid transport demonstrated two-limbed double reciprocal plots suggesting two transport systems for each amino acid. The low concentration limb of the curves established a Km of 0.1 mM for L-leucine and 0.05 mM for L-lysine; Vmax values were 2.0 and 2.85 nmole/mg protein/90 seconds, respectively. Leucine and lysine were shown to be competitive inhibitors of each other. Further competition studies revealed that other amino acids also had affinity for these carriers. Amino acid transport was found to be sensitive to sulfhydryl active compounds. Colchicine treatment of peritoneal macrophages did not inhibit the transport of the amino acids tested. Preloading macrophages with latex beads or heat-killed staphylococci by phagocytosis stimulated 2-deoxy-D-glucose (2-dOG) uptake markedly, but had no measurable effect on amino acid transport. Although total transport of 2-dOG increased in post-phagocytic macrophages, the kinetics of the system were not altered significantly. The Km for both pre- and post-phagocytic transport of 2-dOG was shown to be 1.2 mM and the Vmax was shown to increase from a pre-phagocytic value of 20 nmoles/mg protein/90 seconds to a post-phagocytic 27 nmoles/mg protein/90 seconds. Phagocytosis of heat-killed staphylococci by guinea pig polymorphonuclear leukocytes (PMNs), however, did not cause an augmentation in hexose transport in the cells. The presence of colchicine during phagocytosis did not alter subsequent uptake of amino acids by the macrophages.


Repetitive DNA in differentiating chick tissues.

Embryonic chick DNA from different tissues was examined for differences in relative content of highly repetitive DNA which might indicate specific DNA amplification in somatic cells. The content of repetitive sequences in DNA isolated from cerebrum, muscle, and neural retina tissues, at the same and at different embryonic stages, was determined by hydroxyapatite fractionation of partially reassociated DNA samples. An unrenatured marker DNA (C14-labeled E. coli DNA) was added to each chick DNA sample in order to monitor the nonspecific single-stranded DNA retention by each hydroxyapatite column. When chick DNA samples were sheared to a double-stranded length of 1,300 nucleotide pairs, an average of 20.2% +/- 2.2% of the DNA was found to reassociate at a Cot value of 10. The quantity of the fast reassociating sequences was found to constitute the same fraction of the DNA in all the tissues studied. In addition, all the reassociated DNA samples exhibited the same CsCl density classes. The studies also indicated that most chick DNA repetitive sequences are interspersed with nonrepetitive sequences.


Separation of proximal tubule cells from suspensions of rat kidney cells by free-flow electrophoresis.

Suspensions of rat kidney cells obtained by disaggregation of the kidney with 0.25% trypsin were separated by electrophoresis. Previously, we found a correlation between cells with histochemically demonstrable alkaline phosphatase (HDAP) and cells with brush borders which established that HDAP is a useful marker for rat proximal tubule cells (Kreisberg et al., '77). The starting suspension of cells for electrophoresis consisted of 38.4 +/- 5.7% nucleated cells with HDAP, 39.8 +/- 5.7% nucleated cells without HDAP, and 20.8 +/- 9.2% red blood cells. After electrophoresis, the purest fraction contained 85.8 +/- 3.5% nucleated cells with HDAP, 8.4 +/- 2.2% nucleated cells lacking HDAP, and 5.8 +/- 2.8% red blood cells; 91.9 +/- 2.4% of the nucleated cells in the purest fractions had HDAP.