Connective tissue changes and mast cell variations in benign and malignant lesions of the uterine cervix.

We have studied mast cell variations and stromal changes in 100 benign and 100 malignant lesions of the uterine cervix. Mast cells were found in close proximity to the cervical glands or young proliferating fibroblasts. Comparison of mast cell densities in benign and malignant conditions revealed an increase in inflammatory processes, while in cancers there is diminution in number or total absence of mast cells. There is no conclusive correlation between the age of the patients and the density of these cells. An inverse relationship existed between the mast cell population and degree of anaplasia as well as of mitotic figures.


Seasonal incidence of estrus and interestrous interval for bitches of seven breeds.

For 7 breeds of bitches (Toy Poodle, Cocker Spaniel, Basset Hound, Boston Terrier, German Shepherd Dog, Pekingese, and Beagle), estrous activity occurred in all seasons. Differences in seasonal incidence of estrus were not statistically significant when compared within the total group or within and between breeds. The German Shepherd Dog breed had the shortest interestrous interval (149 +/- 28.5 days), which was significantly different from that of the other breeds. Means interestrous interval in parous Beagle bitches was not significantly different from that in nonparous Beagle bitches. In Beagle bitches, the mean interval between fertile estrous periods was not significantly different from the mean interval between nonfertile estrous periods.


Cytochrome P450-linked p-nitroanisole O-demethylation in the perfused lung.

Oxidative demethylation of p-nitroanisole, a cytochrome P450-linked mixed-function oxidation, was evaluated in isolated perfused rat and rabbit lungs. The product, p-nitrophenol, was monitored continuously in the lung effluent by spectrophotometric measurement. Pulmonary p-nitrophenol production in mumol/h per g dry wt was 6.2+/-0.4 by rabbits and 2.0+/-0.3 by rats (mean+/-SE). Maximal activity of the reaction required pulmonary perfusion rates in excess of 60-80 ml/min per g of dry lung. The half-maximal rate of p-nitrophenol production was observed with p-nitroanisole concentration of 13 micron. Pretreatment of rabbits with chlorpromazine increased p-nitroanisole O-demethylation activity by 63% but phenobarbital pretreatment had no effect. Ventilation with 75% carbon monoxide plus 20% O2 reversibly inhibited the reaction. Specific activity of p-nitroanisole demethylase in the microsomal fraction was 0.5 nmol/min per mg protein in rabbit lungs and 0.1 nmol/min per mg protein in rat lungs. Other rabbit lung subcellular fractions compared with microsomes had significantly lower specific activity. This study demonstrates that p-nitroanisole O-demethylation can be continuously monitored in the intact lung and describes conditions necessary for maximal activity of this pathway.


Survey of 1975 wheat and soybeans for aflatoxin, zearalenone, and ochratoxin.

Wheat samples (102 lots) were collected from Virginia, North Carolina, southeastern Missouri, southern Illinois, and Kentucky. Soybean samples (180 lots) were collected from Virginia, Illinois, Iowa, Minnesota, Nebraska, Alabama, Arkansas, and Texas. Samples of both commodities were analyzed for zearalenone, aflatoxin, and ochratoxin by the Eppley method. None of the 3 mycotoxins was detected in soybeans. Aflatoxins and ochratoxin A were not detected in wheat, but zearalenone was detected in 19 of 42 samples collected in Virginia. Half of the Virginia samples were collected because they were mold-damaged. Zearalenone levels ranged from 0.36 to 11.05 ppm; the identity of the zearalenone was confirmed by gas-liquid chromatography and mass spectroscopy. Gibberella zea infection (6-60%) was detected in all of the zearalenone-positive samples; 6-60% of the kernels in the samples tested contained G. zea.


Rapid digestion and flameless atomic absorption spectroscopy of mercury in fish: collaborative study.

A collaborative study of the determination of mercury in fish has been completed in which wet oxidation of fish tissue in nitric acid, using vanadium as a catalyst, is compared with the AOAC official final action digestion technique, 25.103-25.105, involving a nitric-perchloric acid mixture. The study used tuna fish samples of known mercury content and included spike recovery studies in which methyl mercury solutions of known composition were provided to each laboratory. The study was designed to provide recovery information that bracketed the regulatory level of mercury in fish. The results indicate that the proposed digestion method is at least as precise and accurate as the AOAC method. The proposed method is also more rapid and less hazardous. It has been adopted as official first action.


Collaborative study of gas-liquid chromatographic determination of methaqualone in pharmaceutical and clandestine preparations.

Eight laboratories collaboratively studied a procedure for the quantitative determination of methaqualone. HC1 in pharmaceutical and clandestine preparations. Methaqualone is extracted from an aqueous bicarbonate solution into chloroform and quantitated by gas-liquid chromatography on a 3% OV-1 column. Tetraphenylethylene is used as an internal standard. Two commerical preparations and 4 sample mixtures prepared by the author were studied. Recoveries for the 4 prepared samples ranged from 100.0 to 102.6%, and the coefficients of variation ranged from 1.58 to 4.15% for the 6 samples studied. The method has been adopted as official first action.


Preliminary collaborative study of modified spectrophotometric determination of pyrantel tartrate in swine feeds by the method of standard additions.

A modified spectrophotometric method for determining pyrantel tartrate in swine feeds was subjected to a preliminary collaborative study. Two small-scale commercial pyrantel-medicated feed samples (0.0881 and 0.0106%) were assayed in replicate by 4 collaborators. The mean results of all laboratories were 0.0862 and 0.0112%. The mean coefficients of variation were 10.57 and 6.48%, repectively. Suggestions for improving recovery include the following: complete dissolution of standard, use of analytical grade KI, careful phase separation, thorough mixing, and minimum exposure of compound to light.


High pressure liquid chromatographic determination of sulfanitran and dinsed in medicated feeds and premixes.

A simple reverse phase high pressure liquid chromatographic method is described for determining sulfanitran (acetyl-p-nitrophenylsulfanilamide) and Dinsed (dinitrodiphenylsulfonylethylenediamine) in a variety of feed premixes and formulations. Feed premixes are extracted with dimethylformamide, and formulated feeds are extracted with hot methanol. The extract is filtered through medium porosity paper and injected into a liquid chromatograph equipped with a 254 nm ultraviolet detector and a 30 cm column packed with muBondapak C18. The mobile phase is acetonitrile-water (45 + 55) at a flow rate of 1.0 ml/min. Chromatography was complete in 10 min and peak heights were used for quantitation. Comparison of analyses of commercial samples by this method and by the AOAC colorimetric method, 42.176-42.179, showed close agreement. Recovery of spiked feed samples ranged from 98 to 105%. Butynorate and roxarsone, 2 other drugs which are normally found in combination with sulfanitran and Dinsed, do not interfere.


Extraction, gas-liquid chromatographic detection, and quantitation of hexachlorophene residues from plant tissues high in lipid content.

A method has been developed for the extraction, cleanup, derivatization, detection, and quantitation of hexachlorophene (HCP) residues from 2 types of plant storage tissue high in lipid content. Wet soybean or peanut tissue was homogenized and extracted with ethyl ether and chromatographed on silica gel to remove the neutral lipids. The cleaned up sample was methylated with diazomethane and the dimethoxyhexachlorophene was eluted from a second silical gel column and chromatographed on a 6' glass column packed with 3% OV-1 or 3% SE-30 on Gas-Chrom Q. The instrument detection limit for the 63Ni electron capture detector was less than 0.1 ng for dimethoxyhexachlorophene and about 1 ppb HCP residue in plant issue. Recovery of 10-420 ppb HCP added to tissue averaged 90.9 +/- 5.7%. Interfering substances were removed, column life was increased, peak sharpness was increased, and tailing of the parent compound was decreased by using appropriate column chromatography.


Fluorometric determination of histamine in tuna: development of method.

Tuna extracts are treated with an anion exchange resin to remove interfering materials, histamine is derivatized with o-phthaladehyde, and the fluorescence of the resulting compound is measured fluorometrically. Replicate analyses of acceptable and decomposed tuna packed in oil or water agreed within 1 mg at a level of 10 mg/100 g and within 12 mg at a level of 100 mg/100 g. Recoveries of histamine added to fish were greater than 90 and greater than 83% at levels of 10 and 100 mg/100 g, respectively. The new method is more rapid and specific and is simpler than previous methods because no liquid-liquid extractions or chromatographic separations of histamine are required. The sensitivity of the method allows quantitation of less than 10 mg histamine/100 g sample. The accuracy and precision of the fluorometric method are comparable to those of the official AOAC colorimetric method.


Fluorometric determination of histamine in tuna: collaborative study.

Six samples of canned tuna, albacore, yellowfin, and skipjack, in water or oil pack were analyzed in duplicate by a fluorometric method and the AOAC colorimetric method. For the fluorometric method, recoveries of histamine added to acceptable tuna averaged 99% with a range of 91 to 107%. Agreement between laboratories for the analyses of decomposed tuna containing 20-200 mg histamine/100 g sample was excellent. Results from the fluorometric method are comparable with those from the AOAC colorimetric method; the fluorometric method has been adopted as official first action.


Differentiating the saw-toothed grain beetle, Oryzaephilus surinamensis (L.), and the merchant grain beetle, O. mercator (Fauv.), by morphology of the male genitalia.

Male saw-toothed grain beetles (Oryzaephilus surinamensis) and merchant grain beetles (O. mercator) were dissected, and the major differences in their genitalia were reviewed. Differences in the morphology of the internal sac, setae of the lateral lobes, chitinous rods of the median lobe, and setae on the eighth sternite are illustrated with photomicrographs. The only other difference between the 2 species is the outline of the adult head. The dissection procedure is described briefly.


Calcium-binding subunit of myosin light chain kinase.

Myosin light chain kinase from frozen rabbit skeletal muscle was separated into two protein components by DEAE-cellulose column chromatography. One component (MW 20,000) was a Ca2+-binding protein. Both components and Ca2+ were essential for the enzyme activity.


The complete amino acid seqeunce of mitochondrial asparatate aminotransferase from pig heart.

The complete amino acid sequence of the mitochondrial aspartate aminotransferase from pig heart was determined by analyses of the fragments obtained from tryptic digestion and cyanogen bromide treatment of the protein. The sequence analyzer was useful for establishing the primary structure of the N-terminal portion of the whole protein. There are 401 amino acid residues in the molecule. The sequence was compared with that of the cytoplasmic isozyme, showing 48% homology.


Calcium-sensitivity of the microtubule reassembly system. Difference between crude brain extract and purified microtubular proteins.

Microtubule reassembly in crude extracts of porcine brain was inhibited by 10(-5) M Ca2+, wherease 10(-3) M Ca2+ was required for inhibition of the reassembly from purified microtubular proteins. This accounts for the apparent discrepancies between the results reported by other investigators. Furthermore, the Ca-sensitivity of the purified microtubular proteins was nearly completely recovered on addition of a fraction obtained from crude brain extract.


Nuclear ribonucleoprotein particles in the cellular slime mold Dictyostelium discoideum.

The nuclear ribonucleoprotein (RNP) particles containing rapidly labeled RNA were isolated from interphase cells of the cellular slime mold Dictyostelium discoideum and characterized. The size of the isolated RNP particles was small (10S to 50S) in comparison with that of nuclear RNP particles found in higher eukaryotes. These small RNP particles do not seem to be artifacts due to degradation during the preparation of nuclear extracts. The rapidly labeled RNA of the nuclear RNP particles was heterogeneous in size and a considerable amount contained polyadenylic acid sequences. Synthesis of RNA in the nuclear RNP particles was resistant to a relatively high concentration of actinomycin D. The protein component of the RNP particle consists of at least four proteins with molecular weights of 80,000, 66,000, 60,000, and 42,000. Thus it is suggested that almost all of the nuclear RNP particles containing rapidly labeled RNA in interphase cells are RNP complexes consisting of Heterogeneous nuclear RNA and several protein species.


Trifluoroacetyl tripeptides as potent inhibitors of human leukocyte elastase.

Trifluoroacetylated peptides are much more potent inhibitors of human leukocyte elastase than the corresponding unblocked, acylated or benzyloxycarbonylated peptides. The most active compound was trifluoroacetyl-Val-Tyr-Val (Ki = 1.3 micron). A number of free and NH2-terminal-substituted peptides exhibited similar affinities for porcine pancreatic and human leukocyte elastase, indicating that these two enzymes must have similar specificity sites.


Regulation of chondroitin sulfate synthesis. Effect of beta-xylosides on synthesis of chondroitin sulfate proteoglycan, chondroitin sulfate chains, and core protein.

Monolayer cultures of embryonic chick chondrocytes were incubated with 35SO42- in the presence and absence of 1.0 mM p-nitrophenyl-beta-d-xyloside for 2 days. The relative amounts of chondroitin sulfate proteoglycan and free polysaccharide chains were measured following gel filtration on Sephadex G-200. Synthesis of beta-xyloside-initiated polysaccharide chains was accompanied by an apparent decrease in chondroitin sulfate proteoglycan production by the treated cultures. When levels of cartilage-specific core protein were determined by a radioimmunoassay, similar amounts of core protein were found in both beta-xyloside and control cultures, indicating that decreased synthesis of core protein is not responsible for the observed decrease in chondroitin sulfate proteoglycan production. Activity levels of the chain-initiating glycosyltransferases (UDP-D-xylose: core protein xylosyltransferase and UDP-D-galactose:D-xylose galactosyltransferase) as well as the extent of xylosylation of core protein were found to be similar in cell extracts from both culture types. Furthermore, beta-xylosides did not inhibit the xylosyltransferase reaction in cell-free studies. In contrast, the beta-xylosides effectively competed with several galactose acceptors, including an enzymatically synthesized xylosylated core protein acceptor, in the first galactosyltransferase reaction.


DNA-directed in vitro synthesis of beta-galactosidase. Studies with purified factors.

The phage DNA-directed synthesis of beta-galactosidase has been examined in a system containing the following purified Escherichia coli factors: RNA polymerase; cyclic AMP receptor protein; N10-formyltetrahydrofolate Met-tRNAf transformylase; initiation factors 1, 2, and 3; elongation factors Tu, Ts, and G; release factors 1 and 2; 20 aminoacyl-tRNA synthetases; L factor (Kung, H. F., Spears, C., and Weissbach, H. (1974) J. Biol. Chem. 250, 1556-1562); and Lalpha (Kung, H.-F., Spears, C., and Weissbach, H. (1976) Fed. proc. 35, 1537). Under these conditions, beta-galactosidase synthesis occurs at less than 1% of the rate obtained with unfractionated extracts, which suggested that other required components were lacking. The difficulty in obtaining large amounts of the purified aminoacyl-tRNA synthetases for these studies made it necessary to modify the system. It was possible to conserve many of the purified aminoacyl-tRNA synthetases since at least 13 of them could be replaced by an Ehrlich ascites extract. The ascites extract plus other E. coli purified factors was used as a basic system to search for additional components required for beta-galactosidase synthesis. The present report describes the purification from E. coli extracts of three fractions, called Lbeta, Lgamma, and Ldelta, that are needed to restore enzyme synthesis.


Stress-generated potentials in bone: effects of collagen modifications.

The effect of collagen modification on the magnitude of stress-generated potentials has been studied. A whole bone model has been developed for determining the effects of collagen crosslinking on the electrical signals. Increased crosslinking results in larger electrical signals, and such increases result whether in vivo or in vitro changes in the collagen are made.


The effect of TLCK on transcription and its role in modifying cell growth.

The synthetic protease inhibitor N-tosyl-L-lysine-chloromethyl ketone (TLCK) acts to inhibit transcription when added to cell lines growing in vitro. This inhibition of transcription is most pronounced in transformed cells where TLCK is very toxic at concentrations as low as 25 mug/ml of culture medium. Non-transformed cells are more resistant to the effect of TLCK, requiring ten times more TLCK to produce a comparable inhibition of transcription. The effect of this protease inhibitor on transcription can be prevented by preincubation of the cells in reduced glutathione or cysteine; however, the cells can not be rescued from the effect of TLCK even if glutathione or cysteine are added to the culture medium within five minutes of the addition of TLCK.


Effects of temperature, metabolic and cytoskeletal inhibitors on the rate of BHK cell adhesion to polystyrene.

Adhesion of baby hamster kidney fibroblasts (BHK cells) to a Falcon tissue culture flask was measured under various physiological conditions. While 75-80% of the fibroblasts adhere at temperatures from 19-50 degrees, cellular adhesion decreased dramatically below 19 degrees. Less than 10% of the cells adhere to the substratum even after prolonged incubations at temperatures of 8 degrees or below. This lack of adhesion at low temperatures cannot be overcome by the application of increased gravitational force to the cells. No correlation exists between cellular ATP concentrations or respiration rates and the rate of cell adhesion to the substratum. One millimolar Na F and 1 mM 2,4 dinitrophenol together lower cellular ATP concentration by 95% but adhesion is reduced by only 50%. NaN3 and KCN greatly lower cellular ATP concentrations without a corresponding inhibition of adhesion. Inhibition of cellular respiration by these compounds occurs at lower concentrations than does the inhibition of adhesion. Two micrograms/milliliters of cytochalasin B inhibits adhesion by 90%, 0.1 mM vinblastine sulphate or colchicine by less than 50% and 50 microgram/ml colcemid by less than 30%. Fixing the cells with formaldehyde, hardening their membranes with ZnCl2 or treating the cells with toluene, all cause an inhibition in adhesion. Again, application of increased gravitational force cannot overcome these latter inhibitions of BHK cell adhesion to the surface of the flasks.


Effect of calcitonin on the phosphorylation of non-histones from cultured bone cells.

32P-uptake into non-histones from bone cell cultures was selectively stimulated in the presence of calcitonin. Comparison of the control and experimental radioactivity profiles of non-histones fractionated by SDS gel electrophoresis showed that, in response to calcitonin stimulation, there was a 2- to 3-fold increase in the specific activity associated with non-histone proteins in the molecular weight range of 10,000 to 45,000 daltons while that of bands between 50,000 to 200,000 decreased.


Rapid cell surface-related stimulation of alkaline phosphatase in HeLa cells by dimethyl DL-2,3-distearoylotypropyl-2'-hydroxyethylammonium acetate (Rosenthal's inhibitor).

Cultures of HeLa cells, strains S3G (HeLa65) and S3K (HeLa71) were grown in plastic dishes until firmly attached and were then treated with sonic dispersions of Rosenthal's phospholipase inhibitor. A rapid increase in alkaline phosphatase activity occurred following this treatment in the S3G strain (low inducible alkaline phosphatase) but not in the S3K strain (high constitutive alkaline phosphatase). The stimulatory effect was dependent on the presence of bovine serum in the medium. No stimulation of alkaline phosphatase was observed in a variety of soluble preparations of this enzyme.


Plasma membrane intramembranous particle topography in 3T3 and SV3T3 cells: the effect of cytochalasin B.

The topography of plasma membrane intramembranous particles in contact-inhibited and transformed cells is a topic of current controversy. We have reported that particles are aggregated in 3T3 cells and that they are randomly distributed in SV3T3 cells. Other investigators have, however, proposed that these differences result from the effect of cryoprotectants and are blocked by aldehyde fixation. The experiments described in this paper re-examine PMP topography in large samples of 3T3 and SV3T3 cells prepared by a variety of preparative methods. The results confirm that 3T3 cells contain aggregated intramembranous particles and that native SV3T3 cells do not, regardless of whether cells are prepared in glycerol, sucrose, tissue culture medium or following prefixation in paraformaldehyde. Data presented in this paper also show that cytochalasin B induces dose- and time-dependent disaggregations of particles in contact-inhibited 3T3 cells. Particle disaggregation results from exposure of cells in situ for 1-3 h to 1-500 ng/ml cytochalasin B. Removal of the drug by washing with reincubation of cells in medium containing serum results in particle reaggregation within 1 to 3 h. These observations suggest that intramembranous particles in 3T3 cells are aggregated in the native state and that membrane-active drugs modulate their distribution by a mechanism possibly involving disruption of linkages between membrane components and the cytoskeleton.


A model of pattern formation in insect embryogenesis.

A model is proposed in which the interaction of an autocatalytic substance with a short diffusion range--the activator--and its more diffusible antagonist--the inhibitor--leads to a local high concentration of activator at the posterior pole of the egg. The inhibitor, which is then produced mainly in this activated region, diffuses into the rest of the egg, where it acts as a 'morphogen', forming a concentration gradient which supplies positional information. This model can account quantitatively for the patterns resulting from a large number of different experiments performed during early insect development, including ligation, u.v.-irradiation and microsurgical manipulations. The formation of additional posterior structures is interpreted as the result of the appearance of a new activator peak. Omission of segments after ligation of the egg is explained as the result of accumulation of morphogen (the inhibitor) at the posterior side of the ligation and a decrease of morphogen on the anterior side. In order to account for certain quantitative features of the ligation experiments it is necessary to assume that determination in response to the morphogen gradient is a slow, stepwise process, in which the nuclei or cells first pass through determination stages characteristic for more anterior structures until they ultimately form a given structure. The mutual interactions of activator and inhibitor are expressed as a set of partial differential equations. The individual experiments are simulated by solving these equations by use of a computer.


Mechanochemical coupling in flagella. V. Effects of viscosity on movement and ATP-dephosphorylation of Triton-demembranated sea-urchin spermatozoa.

Improved techniques have been used to measure movement-coupled ATP-dephosphorylation by suspensions of motile, reactivated sea-urchin spermatozoa. The results confirm that the movement-coupled ATP-dephosphorylation is proportional to beat frequency under conditions where the bend angle remains constant. Some observations suggest that the rate of ATP-dephosphorylation is also related to the amplitude of sliding between flagellar tubules, but our attempts to measure this relationship have been unsuccessful.


Studies on the function and composition of the 10-NM(100-A) filaments of vertebrate smooth muscle.

The extraction of isolated vertebrate smooth muscle cells at high and low ionic strength yields cell ghosts which are seen in the electron microscope to be composed of a complex network of 10-nm filaments, together with residual actin. After SDS-gel electrophoresis of the cell ghosts only 2 bands may be recognized, one corresponding to actin and the other migrating at about 55 000 mol. wt that arises from the 10-nm filaments. The 10-nm filaments are extremely sensitive to proteolysis and are absent from cells exposed to crude collagenase in the presence of Triton X-100. Such cells, lacking 10-nm filaments, still contract in response to ATP. The data indicate that the 10-nm filaments are not essential for contraction, but rather form a specialized intracellular cytoskeleton. While completely insoluble in concentrated salt solutions the 55 000 mol. wt protein is readily extracted with acetic acid from homogenized and salt-extracted smooth muscle residue. The extracted protein reassembles, on dialysis, into filaments of about 10-nm diameter and has an amino acid composition almost identical to that deduced for vertebrate neurofilaments. From the cytoskeletal role that the 10-nm filaments play in smooth muscle and, as appears likely, in other cell types the filament protein has been tentatively termed 'skeletin'. Results relating to the proportion of skeletin in smooth muscle and the structure of the 10-nm filaments are described and discussed.


Ultrastructural and radioautographic investigation of the nucleolar cycle in Physarum polycephalum. Characterization of DNA-containing subunits.

The present study has been mainly focused on the nucleolar cycle in the slime mould Physarum polycephalum. The ultrastructural characteristics of the interphase nucleolus, in this species, are quite similar to those of nucleoli in other organisms: it is essentially constituted of large particulate zones surrounding denser regions which are predominantly fibrillar in texture. The latter nucleolar zones, following fixation with osmium tetroxide, are characterized by the presence of opaque granules approximately 25 nm in diameter. Contrary to the situation which generally prevails in other eukaryotes, the late prophase nucleolus fragments into numerous globular bodies which are recognizable by the presence of opaque particles. These fibrillogranular nucleolar fragments persist during mitosis and are observed to become incorporated in the newly formed nucleolus. High-resolution radioautographic observations reveal that these nucleolar remnants contain DNA. The present observations together with recent biochemical data from other authors on the characteristics and mode of duplication of nucleolar DNA in P. polycephalum have led us to the hypothesis that the nucleolus, in this organism, contains several distinct globular subunits each containing ribosomal DNA as a key component. The existence of such morphological subunits appears to account for the unusual behaviour of the nucleolus during the cell cycle.


Separation and biochemical characterization of the two cell types present in the pseudoplasmodium of Dictyostelium mucoroides.

The pseudoplasmodium (slug) of the cellular slime mould, Dictyostelium mucoroides consists of prestalk and prespore cells. These 2 differentiated types of cells were separated by modification of the previous methods using density-gradient centrifugation. Major improvements made in the present study were the use of a density column of different specific gravities and the use of a discontinuous gradient rather than a continuous one. With these improvements, it became possible to obtain efficiently a large number of prestalk and prespore cells. After separation of the 2 types of cells, activities and electrophoretic patterns of some developmentally regulated enzymes were compared. The hydrolases such as beta-glucosidase, beta-galactosidase, acetylglucosaminidase and alkaline phosphatase showed higher activities in the prestalk than in the prespore cells. The results are consistent with the fact that more autophagic vacuoles are present in the prestalk than in the prespore cells. On the other hand, UDP-galactose polysaccharide transferase was almost exclusively found in the prespore cells. Electrophoresis on polyacrylamide gels of slug, prestalk and prespore extracts showed that one among 4 isozymes of beta-galactosidase recognized in the slug extract was present only in the prestalk extract. Electrophoretic patterns of acid phosphatase revealed that one of the two isozymes present in the slug was specifically found in the prestalk cell. Finding of such prestalk specific isozymes was significant, since no specific markers have been known for the prestalk cell.


The structure of nuclear ribonucleoprotein of amphibian oocytes.

Nuclear RNP from Triturus oocytes is organized as strings of beads which can be converted into 20-nm-diameter monoparticles with mild RNase treatment or into 5-nm-thick linear fibrils with low salt treatment. The protein component comprises a heterogeneous size-range of polypeptides which differ from the polypeptides of the other nucleoproteins of oocytes. The RNA is of high molecular weight, sediments mostly in excess of 50 S, and is capable of assuming considerable secondary structure. Duplex regions in the form of hairpin loops are present and may serve as focal points in the condensation of the RNP transcript fibres to generate the periodic beaded structure. The structure of the beads may be maintained by means of protein-protein interaction since at salt concentrations between 1 and 2 M NaC1 all of the proteins are released in a cooperative manner as various sized aggregates which sediment at 15-30 s. There are no specific proteins obviously peculiar to either the beaded or the fibrillar RNP configuration. The various properties of nuclear RNP are compared with those of chromatin.


The isolation of plasma membrane from protoplasts of soybean suspension cultures.

A procedure for the isolation of plasma membranes from protoplasts of suspension-cultured soybean is described. Protoplasts were prepared by enzymic digestion of the cell wall and the plasma membrane was labelled with radioactive diazotized sulphanilic acid. The membrane systems from broken protoplasts were separated by continuous isopycnic sucrose gradient centrifugation. Radioactivity was localized in a band possessing a buoyant density of 1-14 g ml-1. The activities of NADPH- and NADH-cytochrome c reductase, fumarase, Mg2+-ATPase, IDPase and acid phosphodiesterase in the various regions of the density gradient were determined. A plasma membrane fraction was selected which was relatively uncontaminated with membranes derived from endoplasmic reticulum, tonoplasts and mitochondria. The results indicated that Mg2+-ATPase and possibly acid phosphodiesterase were associated with the plasma membrane.


Studies on isolated smooth muscle cells: The contractile apparatus.

Smooth muscle cells may be isolated from the taenia coli muscle of the guinea pig which, when made permeable by treatment with Triton X-100 (0-05%) show a sensitivity to Ca for contraction with MgATP. The rate of contraction, about 10 micron s-1, corresponds closely to the maximum velocity of shortening of the intact muscle. Electron microscopy of such partially demembranated muscle cells shows that myosin filaments of about 16-nm diameter are present in both the rigor and the relaxes states. In addition, the actin and myosin filaments are commonly seen to be associated in groups corresponding approximately in size to the fibrils recognizable in cells in rigor in the light microscope. The dense bodies and the 10-nm filaments are found located between the actin-myosin filament groups. The thick myosin filaments may be isolated by fragmentation of the cells under relaxing conditions. These native filaments range up to about 8 micron in length and show the same structural organization as filaments aseembled from purified smooth muscle myosin: there is no central bare zone and bare edges, about 0-2 micrin long, occur at the filament ends. The lack of bipolarity of the native smooth muscle muosin filaments and the absence, in the contractile apparatus, of actin-associated structures equivalent to Z-lines suggests that the amount of shearing that can occur between the actin and myosin filaments is considerably greater than in skeletal muscle.


Nuclear DNA content and chromosome numbers throughout the life cycle of the Colonia strain of the myxomycete, Physarum polycephalum.

Nuclear DNA content and ploidy have been determined at different stages of the life cycle of the Colonia strain of the myxomycete Physarum polycephalum. Analyses at the plasmodial stage showed that (a) Burton and Fuelgen DNA analyses agreed within 15% with strains which ranged from 0-6 to 3-6 pg of DNA per nucleus; (b) S-phase in Colonia is during the early part of interphase as in the Wisconsin strain; (c) in heterothallic and heterothallic x Colonia crossed strains there are 1-0-1-2 pg of DNA and 70 chromosomes per nucleus and in Colonia 0-6 pg of DNA and 40 chromosomes. Germinating spores of all strains contained one population of cells with about 0-5 pg of DNA and 40 chromosomes and another of larger cells with up to 2-5 pg of DNA and 200 chromosomes. The polyploid nuclei comprised 2-20% of the total in heterothallic strains, 2-65% in heterothallic x Colonia crosses and 25-75% in Colonia. A method was devised for making chromosome spreads of amoebae grown on bacterial lawns. Cells were first exposed to dilute formaldehyde at 26 degrees C for 30 min, then spread on slides with hot lactic acid and strained. Such spreads of CLd (Colonia) and RSD4 (heterothallic) amoebae both contained about 40 chromosomes. The data are consistent with the view that Colonia is haploid throughout its life cycle and that chromosome number is neither halved during sporulation nor doubled during plasmoidal formation. However, the possibility exists that an alternance of ploidy occurs by way of the few diploid nuclei present in the plasmodium.


The nucleolar cycle in man.

Tissue cultures of human embryonal kidney and ovary were examined. In the nuclei of both tissues, one to ten nucleoli have been found. The maximum number of nucleoli is connected with the gene expression of rDNA of the 10 nucleolus organizers of chromosome pairs Nos. 13, 14, 15, 21 and 22, which have secondary constrictions and are the satellite chromosomes in man. The small percentage of cells with 10, 9 and 8 nucleoli is attributed to the rapid association of 3 of the homologous acrocentrics (perhaps of group D). Two of the satellite (SAT) pairs probably associate later after mitosis. The process of fusion is dynamic, resulting in one interphase nucleous--a manifestation of the association of all SAT chromosomes. Dissociation of the nucleolus occurs upon entering prophase, due to the condensation of the chromosomes and retreat of rDNA to the respective secondary constrictions. As a result, the nucleolar number increases again. The pattern of the nucleolar kinetics within the course of one mitotic division is described.


Gene transfer by means of cell fusion I. Statistical mapping of the human X-chromosome by analysis of radiation-induced gene segregation.

Hybrid cells were obtained by virus-induced fusion of hamster cells with irradiated human cells. The analysis of such hybrids permits a study of the effects of lethal doses of radiation on human cells and provides a method of sub-chromosomal genetic mapping that is independent of karyological analysis. Radiation-induced chromosome exchanges are shown to be extremely localized, and a map of 4 X-linked genes is presented.


High-density induction of a quiescent cell state in Physarum polycephalum.

The non-histone protein complement of Physarum polycephalum changes rapidly when microplasmodia are subjected to conditions of high density. The changes in these proteins induced by high density are similar to the changes observed during starvation-induced encystment. A 50% decrease in DNA synthesis, observed after 7 h of starvation, is observed after only 1 h of high density. High density also results in a decrease in RNA synthesis comparable to decreases induced by prolonged starvation. Total heterochromatin increases in response to either high density or starvation. Increased heterochromatization is preceded by an increase in nuclear actin. Mitochondrial morphology and cytoplasmic organization are also similarly altered by starvation and high density. These observations suggest the possibility of a generalized mechanism for cellular transition from active growth to a non-proliferative cell state.


Computer evidence concerning the chemotactic signal in Dictyostelium discoideum.

Observed pulsatile aggregation of cellular slime mould amoebae is simulated on a computer. One spatial dimension is considered. In the simulation, attractant is rapidly secreted by the cells, after a delay period, when a superthreshold attractant concentration is sensed. Cells are refractory to further signals after secretion. Once secreted, the attractant diffuses and is hydrolysed. Movement results if a cell's extending pseudopods sense a supercritical increase of attractant; if increases are sensed on both sides, a sufficiently large difference can also initiate movement. The movement continues for a period independently of further signals, but then can be reversed by a attractant increase (at the back of the cell) that surpasses a high threshold. After 100s, motion stops and the threshold for movement reverts to normal. With the above rules, and with parameter values taken, as far as possible, from the literature, the simulation provides the observed pattern of aggration. Outward moving waves of attractant and organized inward pulsatile 'steps' of cell movement surround a cell that sevretes autonomously every few minutes. Other rules fail to give this picture, or give it only for a relatively narrow range of parameter values. It appears that of the various possible signals for chemotaxos, the most likely to be used by the amoebae is a temporal increase of attractant as sensed by extending pseudopods. Nonetheless, we connot rule out the 'classical' hypothesis that cells directly sense concentration differences.


A study of the arrangement of osteoblasts of rat calvarium cultured in medium with, or without, added parathyroid extract.

The osteoblast layers on the inner surfaces of the half-calvaria of neonatal rats of 2 litters were studied by scanning electron microscopy. Some halves were fixed immediately on dissection. Others were carried through incubation in an Eagle's medium, with or without the addition of parathyroid extract (PTE): for 2, 4, 8 or, most importantly, 24 h. In their initial state, presumed as like the o-h material, the osteoblasts presented as a dense tesselation. After 24 h of culture in the medium, osteoblasts were more sparse in the fields examined and although, in a set of 4 preparations, they retained significant alignment, their orientation had become more random. In addition, they were broader and larger in area. By contrast, with PTE added to the medium, osteoblasts almost maintained their number per field, did maintain their cell area, but became longer and narrower and strikingly parallel in their array. The aligning effect of the PTE preceded, and thus was independent of, its elongating effect.


Gene transfer by means of cell fusion. II. The mapping of 8 loci on human chromosome 1 by statistical analysis of gene assortment in somatic cell hybrids.

A method is described which should permit determination of the order and spacing of genes on all human chromosomes by the analysis of just one set of man-mouse hybrid cells. This method is used to determine the map of 8 loci on human chromosome I. A comparison of the statistical maps of chromosome I and of the X-chromosome with the cytogenetic maps of these chromosomes at metaphase indicates that the statistically derived distances between genes are related to the amount of Giemsa light-band material between the genes.


Rapid micro-method for the measurement of ethchlorvynol in blood plasma and in urine by gas-liquid chromatography.

A rapid gas-liquid chromatographic method has been developed for use in the measurement of the hypnotic drug ethchlorvynol in small (50 microliter) volumes of either blood plasma or urine. Neither solvent transfer nor evaporation steps are used in the procedure and sources of interference have proved to be minimal. The method has been applied primarily to the analysis of specimens obtained from patients who had ingested an overdose of this drug. However with slight modification, the technique may be used in the measurement of the plasma concentrations of ethchlorvynol attained during therapy.


A simple procedure for combined gas chromatographic analysis of neutral sugars, hexosamines and alditols. Determination of degree of polymerization of oligo- and polysaccharides and chain weights of glycosaminoglycans.

A reliable and reproducible method that allows the combined, simultaneous gas chromatographic (GC) determination of neutral sugars, hexosamines, alditols, identification and quantitation of the reducing aldose end-group in oligo and polysaccharides and glycosaminoglycans has been described. It involves the following steps: release of the reducing end-group from its protein linkage in glycosaminoglycans and reduction of this reducing end-group into alditol, release of the components of the reduced polymer by resin-catalysed hydrolysis, nitrous acid deamination of the resin-bound hexosamines in this hydrolysate into anhydroaldoses and a combined derivatization and GC determination of the neutral sugars as aldononitrile acetates, anhydroaldoses as peracetylated oximes and alditols as alditol acetates. Application of the method to determination of degree of polymerization of oligo-and polysaccharides and chain weights of proteoglycans has been described. This method has several advantages over the previous methods.


Quantitative analysis of tricyclic antidepressants in serum from psychiatric patients.

A method for the quantitative analysis of tricyclic antidepressants in the serum of psychiatric patients is described. The method can be used for determining amitriptyline, nortriptyline, imipramine, demethyllimipramine, clomipramine, demethylclomipramine, trimipramine and protriptyline. The method consists in a series of extraction steps followed by gas chromatography with flame-ionization detector. The drugs are determined in their native state. The internal standard method is used for the quantitation.


Analysis of formaldehyde in tobacco smoke by high performance liquid chromatography.

Formaldehyde in tobacco smoke is allowed to react with 2,4-dinitrophenylhydrazine to form the dinitrophenylhydrazone. This derivative is extracted with chloroform and concentrated. The residue in methylene chloride is separated and analyzed by high performance liquid chromatography. A column packed with 10-micron silica is used for separation. The internal standard is triphenylene. This method can also be used for the analysis of other carbonyl compounds in tobacco smoke that react to form hydrazone derivatives.


Pathogenesis of alcohol-induced accumulation of protein in the liver.

Alcohol feeding to rats produced hepatomegaly, associated with enlargement of the hepatocytes. The increase in liver dry weight was accounted for not only by fat but also by protein accumulation, primarily in microsomes and cytosol, with a selective increase in export proteins: concentrations of both immunoreactive albumin and transferrin were augmented in liver microsomes and cytosol of ethanol-fed rats. To investigate the mechanism of this protein accumulation, [14C]leucine was injected intravenously and its incorporation into both liver and serum proteins was measured after various time intervals. Rates of synthesis and export were assessed from protein labeling and specific activities of leucyl-tRNA. Synthesis of liver protein and proalbumin were enhanced by chronic ethanol feeding, but this was not associated with a corresponding rise in serum albumin output. Actually, there was a significant retention of the label in liver albumin and transferrin with delayed appearance in the serum of ethanol-fed rats. This indicated that, regardless of the changes in synthesis, the export of protein from the liver into the plasma was impaired. This alteration in export was associated with a decreased amount of polymerized tubulin in the liver of ethanol-treated animals. Thus, both enhanced protein synthesis and defective export contribute to the ethanol-induced accumulation of liver protein, and the decrease in liver microtubules represents a possible site for impairment of protein export.


Hormonal control of sexual attraction in pseudohermaphroditic female dogs.

Groups of female dogs exposed to different degrees of androgenic stimulation during development and a control group of ovariectomized females were tested for their attraction to tethered male and female stimulus animals. Attraction to the male was measured before and after administration of estradiol, and attraction to an estrous female was tested before and after administration of testosterone propionate (TP). Time spent visiting the tethered male was approximately equal for all groups prior to hormone treatment, but after receiving estradiol, control females exhibited a pronounced increase in visiting time; the second longest visits were paid by females that had received moderate amounts of androgen before birth; more heavily androgenized females exhibited no increase in attraction to the male despite estradiol injections. Visits to the estrous female before administration of TP were longer for some groups than for others, but there was no relation between the degree of perinatal androgenization and mean visiting time. After injections to TP the most pronounced increase in visiting was shown by females that had received the largest amounts of androgen during development, and the second largest increase occurred in the prenatally androgenized group. Control females showed the smallest increase in visiting time.


Gastric secretion and activity-stress lesions in the rat.

In Experiment 1, gastric samples were obtained by means of a pylorus ligation procedure on either the first or last day of the activity-stress procedure. Experimental rats had more stomach lesions and showed a drop in gastric acid on the last day collection. In Experiment 2, rats were surgically prepared with gastric cannulas and pyloric cuffs, and first and last day collections were obtained from the same animal. Experimental activity rats revealed more stomach lesions and a corresponding drop in gastric acid on the last collection than did control rats, which thereby suggests that acid was not a significant etiological variable. The destruction of acid-bearing parietal cells or the back diffusion of hydrogen ions could also explain the low acid values in rats with stomach lesions.


Symmetrical gangrene of the extremities associated with the use of dopamine subsequent to ergometrine administration.

The occurrence of symmetrical gangrene of all 4 extremities after the use of dopamine, subsequent to the administration of egometrine, is described. It is suggested that the concomitant use of these 2 agents should be avoided.


Partial purification of antigens from eggs of Schistosoma mansoni that elicit delayed hypersensitivity.

Schistosoma mansoni egg antigens that elicit delayed hypersensitivity in appropriately sensitized guinea pigs were partially characterized by using ion exchange chromatography and preparative electrophoresis. At least three skin-reactive antigens were found, one of which was purified to homogeneity, as analyzed by polyacrylamide gel electrophoresis (PAGE). This antigen was not adsorbed to CM cellulose, migrated cathodal to guinea pig albumin on electrophoresis, and was adsorbed to DEAE cellulose. A second pool of antigenic activity was obtained by adsorption to CM cellulose and subsequent elution. DEAE cellulose chromatography and preparative electrophoresis of this pool indicated the presence of more than one antigen.


On the mechanism of membrane damage by C: exposure of hydrophobic sites on activated C proteins.

In previous papers we have presented evidence that peptides from C proteins C5b, C7, C8, and C9 become inserted in the lipid bilayer membranes and form a transmembrane channel. Presumably, this insertion follows exposure of hydrophobic domains by C activation. In the present experiments liposomes were made with 14C-phosphatidyl choline (PC) and Forssman antigen in the bilayer, and with 86Rb+ in the aqueous compartments. When such liposomes were incubated with anti-Forssman antibody (A) and guinea pig serum (GPS) as a source of C, substantially more 14C-PC and 86Rb+ were released than from liposomes treated with A and C4-deficient GPS, or with A and heated C, or with C alone, or with A alone. The specific release of PC was dependent on the dose of C. Prior treatment of GPS with cobra venom factor abolished its capacity to release PC. The release of PC by A and C7-deficient human serum (C7D-HS) was the same as that of GPS alone, i.e., there was no specific release. A and C8D-HS produced much less specific release than A and GPS; addition of purified guinea pig C7 or C8 to C7D-HS or C8D-HS, respectively, restored the PC release to its full extent. Hence, part of the PC removal is mediated by C5b,6,7; the remainder is attributable to C8 and/or C9.


Abnormal polyclonal B cell activation in NZB/NZW F1 mice.

Spleen cells from autoimmune (10-mont-old) NZB/NZW (B/W) mice failed to generate appreciable numbers of antibody-forming cells (AFC) in vitro to TNP-substituted sheep erythrocytes in response to the polyclonal B cell activators (PBA), LPS and PPD, despite normal DNA synthetic responses to these agents and normal AFC responses to TNP-Ficoll. The failure to respond to PBA in old B/W mice was not due to suppressor T cells since anti-brain-associated-theta-treated spleen cells still failed to generate AFC in response to PBA. The defect was age-related since cells from young B/W mice generated vigorous AFC responses to PBA. It is suggested that the failure of the spleen cells of old B/W mice to generate AFC is a result of in vitro polyclonal B cell activation in the course of autoantibody formation.


Hydration characteristics and electrical resistivity of stratum corneum using a noninvasive four-point microelectrode method.

We have used a four-microelectrode probe to study the relationship between electrical resistivity (p), water content (wf) water binding, and electrolyte levels in samples of plantar stratum corneum. Untreated samples characteristically showed an exponential relationship between log p and wf. This characteristic was retained but altered quantitatively by factors which increase and decrease the ratio of bound to unbound water, by lipid extraction, and (following extraction) by electrolyte presoak. The theoretical aspects of obtaining similar data from intact stratum corneum are discussed.


Streptococcal M protein: an antiphagocytic molecule assembled on the cell wall.

After extraction with nonionic detergent, type 6 streptococcal M protein was found to be composed of multiple proteins ranging in molecular weight from 35,000 to 6,000 daltons. The antiphagocytic proteins, however, were found to be limited to three species having molecular weights of 28,000, 31,000, and 35,000 daltons. These molecules which removed opsonic antibodies from immune serum could be separated from those proteins that had only type specificity. Pulse chase experiments supported by chemical and immunological data suggest that the smaller, type-specific molecules are used to assemble the larger, antiphagocytic proteins. Type 6 M protein was radialabeled and used in a binding assay for the measurement of opsonic antibodies in human serum. Good correlation was observed between binding and the presence of opsonic antibodies in both systems. However, certain sera did exhibit binding but lacked type-specific opsonic activity. Results of competitive inhibition experiments demonstrated that the nonopsonic serum was deficient in certain antibodies that were present in opsonics serum and that the anitphagocytic molecules contained the sites necessary to bind these antibodies.


Capillary blood flow in the genital tracts of conscious ewes: cyclic changes and the effect of ovarian hormones.

The indicator fractionation technique with [86Rb]rubidium chloride as the indicator was used in conscious animals to determine the relative blood flow (RBF) as a measure of capillary blood flow in regions of the genital tract of cyclic ewes and of hormonally treated ovariectomized ewes. The RBF values in the ampulla, isthmus, uterus and cervix of cyclic ewes were markedly raised at pro-oestrus and declined thereafter to low levels during the remainder of the oestrous cycle. On Day 4 a small secondary rise in flow occurred in the ampulla and isthmus. In ovariectomized ewes, oestradiol induced high levels of RBF in the ampulla, isthmus, uterus and cervix whether the animals had been treated previously with progesterone or given concurrent small doses of progesterone. Though progesterone given alone caused a significant rise in RBF in the uterus, this effect appeared to be one occurring only during active re-growth of regressed tracts. The findings are discussed in relation to oestradiol inducing a stimulatory effect on RBF in the genital tract of the ewe around oestrus but not at Days 3-4 of the oestrous cycle.


Binding of methyltrienolone (R1881) to a progesterone receptor-like component of human prostatic cytosol.

The synthetic steroid methyltrienolone (R1881, 17beta-hydroxy-17alpha-methyl-estra-4,9,11-trien-3-one) binds with high affinity to protein in cytosols prepared from human hyperplastic prostate. R1881 also binds to the androgen receptor of rat prostate and the progesterone receptor of rabbit uterus. Other steroids compete with R1881 for unoccupied binding sites in the human prostatic cytosols in a manner similar to that observed with the rabbit uterine progesterone receptor, rather than the rat prostatic androgen receptor. The progesterone receptor-like binding sites are a feature of the prostatic stroma rather than the epithelium.


Sensitivity of anterior hypothalamic areas to gonadal steroid implantation in androgenized female rats.

A single injection of 1 mg of a complex of testosterone esters on day 5 of life was used to prepare constantly oestrous rats. Such androgenized female rats were then ovariectomized and submitted to stereotaxical implantation of 1 microgram oestradiol benzoate, 5 microgram testosterone isobutyrate or, as a control, 10 microgram cholesterol in the anterior hypothalamic areas. The effects of the steroids on plasma and pituitary FSH and LH were assessed by radioimmunoassay. As reported previously by us in normal female and male rats, the preoptic-suprachiasmatic area (POA) was able to control synthesis and secretion of both gonadotrophins and did not lose its sensitivity to oestradiol and testosterone in andorgenized rats. Evidence for enhanced prolactin secretion in androgenized rats was derived from immunofluorescence studies of the pituitary gland and from histology of the mammary glands. In this respect the condition of the androgenized females was opposite to that of the males. The present work demonstrated that stimulation of prolactin secretion in androgenized female rats resulted from oestrogen action due to permanent oestrus rather than from impairment of hypothalamo-hypophysial relationships. Indeed, prolactin stimulation was suppressed when the androgenized rats were ovariectomized and restored when they were subsequently implanted with oestradiol in the POA.


Oscillating contractions in protoplasmic strands of Physarum: simultaneous tensiometry of longitudinal and radial rhythms, periodicity analysis and temperature dependence.

1. The construction of a 'twin-tension transducer' allows the simultaneous measurement of the same or different contraction rhythms at any selected sites of living plasmodia of Physarum polycephalum. This method has been used to analyse the relation of longitudinal and radial contraction activity within migrating plasmodia and plasmodial veins, under isometric as well as under isotonic conditions of measurement. 2. A periodicity analysis of the oscillating contraction rhythms revealed average period values for the longitudinal contraction cycle of 2-1 min and for the radial contraction cycle of 1-3 min at a temperature of 22 degrees C. 3. The periods of longitudinal contraction depend on the environment of the strands. The mean value under submerged conditions was 2-9 min. 4. The temperature dependences for both longitudinal and radial contraction cycles were determined to provide reliable values for the normal reaction range of the contractile system (cytoplasmic actomyosin). The values for radial contraction activity are 2-0 min at 16 degrees C, 1-5 min at 20 degrees C, and 1-2 min at 24 degrees C. The range between 16 degrees and 24 degrees C can be regarded as physiological. 5. The possibility is discussed that only one 'genuine' contraction frequency of cytoplasmic actomyosin exists in Physarum.


Biosynthesis of pro-C3, a precursor of the third component of complement.

A precusor of the third component of complement, pro-C3, was detected in studies of cell-free synthesis and intracellularly in homogenates of liver tissue cultures. The molecular weight of pro-C3 was indistinguishable from that of intact native C3 secreted in vitro by liver or peritoneal macrophages, but its structure was different. Pro-C3 is a single polypeptide chain, whereas C3 secreted by cells in culture consists of two polypeptide chains (mol wt 120,000 and 76,000) linked by disulfide bonds.


Cellular and genetic control of antibody responses in vitro. II. Ir gene control of primary IgM responses to trinitrophenyl conjugates of poly-L-(Tyr,Glu)-poly-D,L-Ala--poly-L-Lys and poly-L-(His,Glu)-poly-D,L-Ala--poly-L-Lys.

The in vitro primary IgM anti-hapten responses to trinitrophenyl (TNP) conjugates of poly-L-(Tyr,Glu)-poly-D,L-Ala-poly-L-Lys (T,G)-A--L and poly-L(His,Glu)-poly-D,L-Ala--poly-L-Lys (H,G)-A--L were shown to be T-cell dependent and under autosomal dominant H-2-linked Ir gene control which mapped within the K or I-A regions of the H-2 complex. The in vitro response to TNP-keyhole limpet hemocyanin, while T-dependent, was not under demonstrable genetic control. The genes governing the in vitro primary IgM anti-hapten responses to TNP-(T,G)-A--L and TNP-(H,G)-A--L resemble the Ir genes controlling the in vivo secondary IgG responses to (T,G)-A--L and (H,G)-A--L in that they are autosomal dominant, map identically within the H-2 complex, and have identical responder and nonresponder haplotypes. It is concluded that Ir genes can govern the ability to generate an IgM response upon initial exposure to antigen.


Synthesis and distribution of H-2 antigens in preimplantation mouse embryos.

Synthesis of H-2 antigens by preimplantation mouse embryos is first detectable at the late blastocyst stage. These antigens were detected using immune precipitation assays of extracts of embryos labeled by incorporation of radioactive amino acids but not by surface iodination. Experiments using isolated inner cell massess and trophoblast vesicles indicate that it is the cells of the inner cell mass that synthesize these antigens. H-2 antigens were not detected in either early blastocysts or at earlier cleavage stages.


Nucleic acid synthesis in preimplantation rabbit embryos. III. A "dark period" immediately following fertilization, and the early predominance low molecular weight RNA synthesis.

This report stems from efforts to characterize the products of transcription during the initial stages of cleavage in the rabbit embryo, since previous studies have suggested that RNA synthesis is required during this period for the continuation of normal development. Early cleaving embryos were exposed to radiolabeled precursors of nucleic acids in vitro and RNA extracted and analyzed by polyacrylamide gel electrophoresis and BD-cellulose chromatography. No unequivocal evidence of RNA synthesis could be obtained during the first three cleavage divisions; all apparent "embryonic" RNA synthesis could be ascribed to contaminating follicle cells. After the fourth cleavage, however, there is readily detectable incorporation of radioisotope into RNA by embryos which have been denuded of follicle cells. Two-thirds of the cytoplasmic RNA labeled during a 4-hour period is 25-30 X 10(3) daltons in size, and some of this low molecular weight RNA can be aminoacylated and shown to be transfer RNA. A low level of ribosomal RNA synthesis is detectable at this stage. The contribution, if any, of the rabbit embryonic genome to development prior to the 16-cell stage remains to be defined.


The estrous cycle in golden hamsters: a circadian pacemaker times preovulatory gonadotropin release.

In a population of cycling female hamsters entrained to an LD 6:18 light cycle (lights 1000-1600 hours), preovulatory release of luteinizing hormone and follicle-stimulating hormone occurred in some animals at 1300-1400 hours and in others at 1900 hours. In every case peak release was phase-locked (2-3-hour positive phase angle) to the circadian rhythm of locomotor activity. The pattern of entrainment of gonadotropin release on LD 6:18 is fully explicable in terms of the hamster's phase response curve to light. We conclude that periodic gonadotropin release in cycling females is timed by a circadian oscillator (biological clock) that is probably the same oscillator driving the circadian rhythm of locomotor activity.


Homozygous mouse embryos produced by microsurgery.

Homozygous mouse blastocysts have been produced following microsurgical removal of one pronucleus from the fertilized egg, diploidization in cytochalasin B, and culture in vitro. This combination of techniques should greatly shorten the time required to obtain homozygous strains of mice.


A reassessment of decreased amino acid accumulation by Ehrlich ascites tumor cells in the presence of metabolic inhibitors.

This study was undertaken to examine the mechanism by which metabolic inhibition reduces amino acid active transport in ehrlich ascites tumor cells. At 37 degrees C the metabolic inhibitor combination 0.1 mM 2,4-dinitrophenol (DNP) + 10 mM 2- deoxy-D-glucose (DOG) reduced the cell ATP concentration to 0.10- 0.15 mM in less than 5 min. This inhibition was associated with a 20.6 percent +/- 6.4 percent (SD) decrease in the initial influx of alpha-aminoisobutyric acid (AIB), and a two- to fourfold increase in the unidirectional efflux. These effects could be dissociated from changes in cell Na(+) or K(+) concentrations. Cells incubated to the steady state in 1.0-1.5 mM AIB showed an increased steady-state flux in the presence of DNP + DOG. Steady- state fluxes were consistent with trans-inhibition of AIB influx and trans-stimulation of efflux in control cells, but trans- stimulation of both fluxes in inhibited cells. In spite of the reduction of the cell ATP concentration to less than 0.15 mM and greatly reduced transmembrane concentration gradients of Na(+) and K(+), cells incubated to the steady state in the presence of the inhibitors still established an AIB distribution ration 13.8 +/- 2.6. The results are interpreted to indicate that a component of the reduction of AIB transport produced by metabolic inhibition is attributable to other actions in addition to the reduction of cation concentration gradients. Reduction of cell ATP alone is not responsible for the effects of metabolic inhibition, and both the transmembrane voltage and direct coupling to substrate oxidation via plasma-membrane-bound enzymes must be considered as possible energy sources for amino acid active transport.


Anomalous permeabilities of the egg cell membrane of a starfish in K+-Tl+ mixtures.

The electrical properties of "inward" rectifying egg cell membranes of the starfish mediastera aequalis have been studied in the presence of K(+)-Tl(+) mixtures. When the ratio of the external concentrations of these ions is changed while their sum is kept constant, both the conductance and the zero-current membrane potential go through a minimum, showing clear discrepancies from theoretical results based on conventional electrodiffusion models (E.g., Goldman's equation). By contrast, when the ration of the two concentrations is fixed and their sum varied, the potential follows an ideal Nernst slope, consistent with Goldman's equation. The membrane conductance which, according to previous studies on similar membranes, is to be viewed as a function of the displacement of the membrane potential from its resting value deltaV, shows marked differences between the cases in which K(+) or Tl(+) are the predominant ions: when K(+) is the predominant permeant ion in solution, the addition of small amounts of Tl(+) inhibits the current, while corresponding blocking effects of K(+) on the current are not observed when Tl(+) is the predominant permeant ion. Also, the time course of the conductance during voltage clamp is different in the two cases, being much faster in Tl(+) than in K(+) solution for comparable values of deltaV. Most of the above features are accounted for by a model in which it is assumed that the ionic channels have external binding sites for cations and that their permeability properties depend on the species of the cation bound (K(+)or Tl(+) in the present experiments).


Characterization of virus-specific messenger RNAs from avian fibroblasts infected with fowl plague virus.

In cell-free protein synthesizing systems from wheat embryos, messenger RNAs extracted from chick embryo fibroblasts infected with fowl plague virus direct the synthesis of nine virus-specific polypeptides, two of which may be related to the virus-specific glycopolypeptides. All of the mRNAs are complementary in sequence to virion RNA, and RNAs which do not contain poly A appear to be translated as efficiently as their polyadenylated counterparts. Under certain conditions of incubation, virion RNA also directs the synthesis of discrete polypeptides but these products are not detected in infected cells.


Amantadine-resistance as a genetic marker for influenza viruses.

The infection of eggs, cell cultures or mice with a mixture of amantadine-resistant and amantadine-sensitive strains of influenza virus resulted in the transfer of amantadine-resistance or sensitivity between strains. The response of a recombinant virus to amantadine was not related to either of its surface antigens. Resistance to amantadine was transferred as an all-or-none character. It is concluded that amantadine-resistance is a useful genetic marker for influenza viruses.


Failure of the RA 27/3 strain of rubella virus to induce intrinsic interference.

Unlike other strains tested, the RA 27/3 vaccine strain of rubella virus, attenuated in human fibroblasts, failed to inhibit superinfection with Newcastle disease virus (i.e. induce intrinsic interference). Since proteins of the input virion are known to lead to intrinsic interference, these may differ in the RA 27/3 strain from those in other rubella strains.


Sensory factors which affect the acceptance of raw and heated defatted soybeans by rats.

The acceptability of heated soybean meal (HS) compared to raw soybean flakes (RS) was studied by means of diet selection in free choice situations. The selection by rats of diets containing HS over those containing RS was shown in short- and long-term preference tests. Texture did not seem to be an important factor in the acceptability difference between HS and RS. Rats selected a less nutritious diet containing RS + sodium saccharin (appealing taste) over a diet containing HS + sucrose octaacetate (aversive taste) for an initial 6 to 7 days, suggesting that post-ingestional factors did not markedly affect the acceptability during that period. However, after this initial period, the rats selected the diet of higher nutritive value, even though it had added sucrose octaacetate. Preference tests before and after producing anosmia indicated that RS contains an aversive odor(s) which could be removed by vacuum treatment. Heated soybean meal was found to contain an appealing taste stimulus (i) which could also be removed by vacuum treatment.


Colitis, persistent diarrhea, and soy protein intolerance.

Four infants with suspected soy protein intolerance were studied prospectively. All had vomiting, diarrhea, hematochezia, and weight loss of more than 10%. Laboratory abnormalities included anemia (3), acidosis (3), hypoalbuminemia (3), hypernatremia (1), hyponatremia (3), and mild eosinophilia (2). Symptoms developed between three hours and five days after systematic testing with increasing volumes of soy formula. After challenge with soy protein formula, proctosigmoidoscopy and rectal biopsy were performed within 24 hours of developing stools with gross blood. Proctosigmoidoscopy showed both spontaneous and induced friability and loss of vascular pattern in mucosa. Rectal biopsy showed acute colitis with crypt abscesses, depletion of mucus from rectal glands, and polymorphonuclear leukocytes within the lamina propria. Soy protein-induced colitis should be suspected in those infants who present with persistent and bloody diarrhea after being fed soy formula.


Congenital thyroid cyst of ultimobranchial gland origin.

The ultimobranchial gland (UBG) is an endocrine gland which contributes C cells to the thyroid gland in mammals. Dysembryonic tissues derived from the UBG have been described as cystic bodies. The following case illustrates the criteria which characterize dysembryogenesis of the UBG.


Plant anticancer agents III: Isolation of indole and bisindole alkaloids from Tabernaemontana holstii roots.

Certain active antileukemic and cytotoxic fractions prepared from Tabernaemontana holstii roots were investigated, resulting in the isolation of the known indole alkaloids conoduramine, conodurine, coronaridine, gabunine, 19-oxocoronaridine, pericyclivine, perivine, and vobasine. Two new alkaloids were assigned the structures 19-oxoconodurine and 19-(2-oxopropyl)conodurine. Both gabunine and 19-(2-oxopropyl)conodurine showed significant inhibitory activity against P-388 cell culture. All of the alkaloids are reported for the first time from T. holstii; conodurine, conoduramine, gabunine, perivine, and pericyclivine are reported for the first time from any Tabernaemontana species.


Antitumor agents XXIII: Helenalin, and antitumor principle from Anaphalis morrisonicola HAY.

A chloroform extract of the whole plant of Anaphalis morrisonicola HAY, showed significant antitumor activity against Ehrlich ascites carcinoma, Walker 256 carcinosarcoma, and P-388 lymphocytic leukemia. Systematic fractionation of the extract led to isolation and characterization of helenalin, a pseudoguaianolide, as the major active principle.


Psoriasis associated with anterior uveitis.

A patient with psoriasis and anterior uveitis is described. In her family the two entities of iridocyclitis and psoriasis occurred separately in two generations and appeared associated in the third generation. Tissue typing of the family revealed HL-A 27 antigen in two members and HL-A 17 in the other two members. The possible relevance of HL-A antigens to the two clinical entities is discussed.


A twin study of temperament in young children.

Temperament was assessed in 182 young children (91 pairs of twins, average age of 3.6 years) with use of the Colorado Childhood Temperament Inventory (CCTI). The CCTI represents a merger of dimensions suggested by two approaches to the study of the development of temperament: the New York Longitudinal Study and Buss and Plomin's temperament theory. Of the six traits measured by the CCTI, genetic factors were implicated in the etiology of five (sociability, emotionality, activity, attention span-persistence, and soothability). One trait, reaction to food, showed no genetic influence and indicated strong between-family environmental effects. A negative correlation between emotionality and soothability may also have a genetic basis. These results suggest that the development of certain personality traits--those that we include as temperament--are substantially influenced by heredity.


Prevalence of venereal disease in indigent pregnant adolescents.

The prevalence of venereal disease in an indigent adolescent maternity population was determined. Of 740 teenagers examined for gonorrhea and syphilis, 2,4% were found to have gonorrhea, whereas 0.6% had primary syphilis. A concomitant study of 740 adult maternity cases at the same hospital revealed a somewhate higher incidence. Some factors in venereal disease prevalence in pregnant teenagers are discussed briefly.


Synthesis and biological activity of 8-arylergolines.

9,10-Didehydro-6-methyl-8beta-arylergolines 2, in which the carboxyl group of lysergic acid and isolysergic acid is replaced by various aryl groups, were prepared in two steps by alkylation of aromatic substrates with the tetracyclic allylic alcohol 3, followed by aromatization with MnO2. The new ergolines 2 have modest prolactin-inhibiting and rat antimuricidal activities and possess significant alpha-blocking and antiserotonin properties.


Intercellular communication and tissue growth: IX. Junctional membrane structure of hybrids between communication-competent and communication-incompetent cells.

The structure of the membrane junctions of the hybrid cell system, examined in the companion paper in respect to competence for communication through cell-to-cell membrane channels, is here examined by freeze-fracture electron microscopy. The junctions of the channel-competent parent cell and of the channel-competent hybrid cells present aggregates of intramembranous particles typical of "gap junction"; those of the channel-incompetent parent cell and channel-incompetent segregant hybrid cells do not. Competence for junctional communication and for gap junction formation are genetically related. The junctions of the intermediate hybrid cells with incomplete channel-competence (characterized by cell-to-cell transfer of small inorganic ions but not of fluorescein), present special intramembranous fibrillar structures instead of discrete gap-junctional particles. The possibility that these structures may constitute coupling elements with subnormal permeability is discussed in terms of incomplete dominance of the genetic determinants of gap junction.


Role of extracellular virus on the maintenance of the persistent infection induced in Aedes albopictus (mosquito) cells by Sindbis virus.

Sindbis virus infection of cultured mosquito cells was found to have no effect on the growth of these cells; instead, a persistent infection of the culture followed an initial acute phase of rapid virus synthesis. Nearly all of the cells in the acute stage of infection were found to actively release virus in an infectious-center assay and to contain significant amounts of virus antigen as determined by immunofluorescence. Cells in the persistent phase of infection released few virions into the media, and only a small percentage of the cultured cells could be demonstrated to contain detectable amounts of virus antigen by immunofluorescence assay. In spite of the fact that nearly 100% of the cells in the persistent phase of infection were found to be virus negative by the two assays described above, the culture as a whole totally excluded the expression of superinfecting virus, as did cells in the acute phase, suggesting that most of the persistently infected cells did, indeed, contain virus information. Prevention of reinfection of the cells in the persistent phase by eliminating extracellular virus resulted in a curing of the culture such that it responded to infection by added virus much as would an uninfected culture.


Mechanism of murine leukemia virus-induced fusion in rat myoblasts defective in differentiation.

fu-1 cells, a line of rat myoblasts defective in differentiation, can be fused into multinucleate syncytia by Moloney murine leukemia virus. The effects of treating the virus with specific antibody, UV irradiation, and elevated temperature and the requirements for cellular RNA and protein synthesis have been studied as they relate to this virus-induced fusion. The results indicate that intact, but not necessarily infectious, virions are required to promote fusion of fu-1 cells. Neither actinomycin D nor cycloheximide altered the formation of syncytia; thus, neither viral nor cellular RNA or protein synthesis is required for fusion. fu-1 cells infected with the ts3 temperature-sensitive mutant of Moloney murine leukemia virus accumlate large amounts of budding  virus on their cell membrane; however, this membrane-associated virus failed to induce syncytia. Upon release of the virus at the permissive temperature, fusion did occur. We conclude that contact or attachment of the immature virus to the cell membrane is not sufficient to promote murine leukemia virus-induced cell fusion; complete virions are required. From these data, we propose that adsorption and penetration of the virus may induce a change in the cell membrane that subsequently promotes the fusion of susceptible cells.


Effect of interferon on dimethyl sulfoxide-stimulated Friend erythroleukemic cells: ultrastructural and biochemical study.

Treatment of dimethyl sulfoxide-stimulated Friend erythroleukemic cells (clone 745) with mouse interferon (50 U/ml) led to the following changes: (i) a net decrease (40 to 60%) in both the total number of apparently newly synthesized virion particles per cell section and in the average number of cell sections containing one or more virion particles; (ii) a large decrease (80 to 90%) in the number of particles released into the supernatant fluid, as assayed by reverse transcriptase activity; (iii) an initial increase in the number of "immature" or "enveloped A-type" virions followed by an increase in the accumulation of empty, core shell-like particles; and (iv) a decrease in the number of cytoplasmic vacuolar structures, which have been implicated as a major site of virus production and which we show here by serial sectioning to be, in several instances, invaginations of the plasma membrane. The effects on virus production were noticeable 2 h after interferon addition and reached their full extent by 13 h. We conclude from these observations that interferon acts upon the late stage(s) of virion maturation, leading both to a decrease in virion production as well as to the formation of defective particles. In contrast, a small but significant increase in the rate at which globin mRNA and hemoglobin accumulate is observed after interferon treatment.


Synthesis of influenza virus polypeptides in cells resistant to alpha-amanitin: evidence for the involvement of cellular RNA polymerase II in virus replication.

Influenza virus polypeptides were not synthesized in wild-type CHO-S-infected cells in the presence of alpha-amanitin, but were synthesized in CHO-Amal cells, a mutant cell line whose DNA-dependent RNA polymerase II is specifically resistant to this drug, indicating that this cellular enzyme is involved in influenza virus replication. The results of experiments designed to detect viral polypeptides synthesized from primary transcripts suggest that the synthesis of a cellular RNA species by RNA polymerase II is required for primary transcription of the influenza virus genome.


The bacterial flora of the vaginal vestibule, urethra and vagina in the normal premenopausal woman.

A study of 100 healthy, premenopausal women demonstrated that lactobacilli and staphylococci are the predominant normal bacterial flora in the vaginal vestibule, urethra and vagina, with diphtheroids, streptococci and micrococci occurring in decreasing order. Gram-negative enterobacteria are a rare occurrence in the normal bacterial flora of the vaginal vestibule (7 per cent), urethra (9 per cent) and vagina (6 per cent). A second study of 10 healthy premenopausal women in whom serial examinations were done confirmed the findings of the first study and revealed that when gram-negative enterobacteria do colonize the normal vaginal vestibule, urethra and vagina colonization is of a transitory nature since enterobacteria tend to disappear spontaneously.


[Correlation between the indicators of blood coagulation and lipid metabolism in patients with stenocardia during the 24-hour period].

The correlation between the main parameters of blood coagulation and lipid metabolism were examined dynamically during 24 hours in 49 patients with angina pectoris. Circadian fluctuations of varying degrees of direct and inverse correlation were found in the concentration of cholesterol, beta-lipoproteins, non-esterified fatty acids, lecitin, total lipids, triglycerids, on the one hand, and the level of free heparin, the fibrinolytic activity, fibrinogen, thrombine time, platelets count in 1 mm3 of blood, on the other hand.


[The indications for the preventive treatment of retinal detachment (author's transl)].

The author describes the genuine indications for the preventive treatment of retinal detachment. He was above all guided by the minute observations of the eye fundus of two eyes with the Goldmann-glass. The author's long experience and his auto-observation of the retinal detachment operation and of preventive treatment show the confidence which can be given to the ideas published in this article.


[Retinopathy in the course of a homozygous haemoglobinopathy (author's transl)].

Fundus examination of a young African female affected with acute benign unilateral iridocyclitis showed lesions similar to those of sickle cell disease. She had a homozygous C/C hemoglobinosis, which was previously unknown. Certain hematological lesions confirmed the pathogenesis of these pathological entities in absence of sickle cell formation.


[Experience with cryotherapy of iridocyclitis (author's transl)].

As a method of treating iridocyclitis, cryotherapy, combined with the universally accepted systemic and topical treatment, is becoming important since it is able to alleviate the symptoms and to speed up the healing of the inflammatory condition of the iris and ciliary body.


[Simultaneous occurrence of juvenile diabetes and Addison's disease (author's transl)].

The occurrence of diabetes mellitus and Addison's disease together is discussed by the authors. The female patient described here had diabetes at the age of four; Addison's disease appeared when the patient was 19 years old. Four years prior to the appearance of Addison's disease, an ophthalmoscopic examination revealed microaneurysms. The examination of the fundus of the eye was negative when Addison's disease, was diagnosed. An HLA-B8 antigen positivity was established. The authors discussed the pathogenetic relationship of HLA-B8 antigen positivity, diabetes mellitus, and Addison's disease.


[Transcutaneous PO2 measurement during balloon atrioseptostomy in an infant with transposition of great vessels].

During ballonatrioseptostomy PO2 was measured transcutaneously and continuously. This allows to control the increasing of the left to right shunt at atrial level and other changes of haemodynamic.


[Timing of a letal intracranial hemorrhage by means of the HbA/HbF ratio (author's transl)].

Possibilities and limits of the method for timing an intracranial hemorrhage by means of the HbA/HbF erythrocyte ratio are investigated in six newborns. The effect of labeling HbF erythrocyte containing newborn blood by transfusion and exchange transfusion of HbA erythrocyte containing donor blood as well as "self-labeling" after the first transfusion as a result of HbA--and HbF erythrocytes decaying at a different rate, is described. If a coagulum can be clearly allocated to the blood where it originates from, depends a) on the alteration of the HbA/HbF erythrocyte ratio by the above mentioned ways of labeling and b) on the size of the sample i.e. on the number of erythrocytes counted to determine the HbA/HbF erythrocyte ratio. The confidence limits of the HbA/HbF erythrocyte ratio for a given number of counted erythrocytes have been determined. It was taken into account whether the blood was labelled by a transfusion or by "self-labeling". The analysis of the period before, during and after the intracranial hemorrhage made a preliminary evaluation of etiological factors and some symptoms possible. The hypothesis that sodium bicarbonate dosages of 8 mEqu/kg/24 h are an important cause of intracranial hemorrhage has to be rejected on the grounds of this study's results.


[Agammaglobulinemia and secretory IgA (author's transl)].

After serum IgA has been intravenously administered, it appears immediately in secretions, sputum, and urine. At about the same time, a mechanism is employed which leads to the concentration of IgA in the sputum. In urine, 7sIgA and SC were separately eliminated; a linkage with 11sIgA did not occur. Similar relationships are probably present in the sputum, but exact proof could not be provided. By administrating immunoglobulin concentrates which contain IgA in 7s-serum form, the IgA concentration in external secretions can be raised; replacing the physiological 11s-IgA however, is not possible. The extent to which 7sIgA can take over the function of 11sIgA in these secretions has not yet been clarified. In this connection, the use of local 11sIgA inhalation therapy rather than ultrasound atomazation seems to be well founded theoretically and also promising. Whether such therapy can be carried out practically and is successful remains to be seen.


[Xanthogranulomatous pyelonephritis in a young child (author's transl)].

Case report on a boy aged 4. The whole of the right kidney showed changes of xanthogranulomatous pyelonephritis. dense granulomatous infiltrations of xanthome cells, epitheloid and plurinuclear giant-cells, lymphocytes and plasma cells. Diagnostic proof was based on the xanthome cells, large histocells with partially pycnotic nucleus and wide margin of cytoplasm. This condition has, so far, been seen only very rarely in children.


[Tri-iodine-thyronin (T3)-hyperthyreosis (author's transl)].

Report on a girl aged 12 years and 2 months and a boy aged almost 14. The first always had normal T4- with always raised T3, belongs, therefore, to the entity T3-hyperthyreosis described by Hollander in 1968. The boy had clear clinical signs of hyperthyreosis, but was untreated for 14 months, because a nuclear medical institute had found T4 to be normal and did not investigate T3. One suspects that proper treatment would have been given, if T3 would have been examined early. Discussion of the entity: T3 hyperthyreosis and of the circumstances under which T3 may be found to be raised even in the absence of the disease.


Discrete subvalvular aortic stenosis. An evaluation of operative therapy.

Angiocardiographic and operative observations support the validity of classifying the spectrum of congenital subvalvular aortic stenosis into a membrane, fibromuscular collar, and tunnel. Our current operative method is to excise a thin membrane or thick fibrous ridge, and, if a fibromuscular collar or tunnel is identified, to effect a left ventricular myomectomy as described by Morrow for hypertrophic subaortic stenosis. Data from experience with 35 children indicate that this approach is effective and safe. Gradients are substantially reduced and residual obstruction acceptable. Successive clinical evaluations (100 percent of 33 survivors) over an interval of 1 to 13 years (mean of 6) affirm that amelioration of the obstruction endures.


[Epidemica of hand-foot and mouth disease].

After describing the affections caused by the virus Coxsackie, the characteristics of disease in mouth-hands-feet are looked over reporting the principal epidemic outbreak as well as the isolated forms of exhibition. The illness can be shown in a complete, incomplete or moderate way. This outbreak occurred in Mendoza (Argentina) in 1975-1976: 25 school aged children were controlled and no other relatives of different ages fell ill. No predisposition for either sex. They didn't come from the same school. A sudden eclosion and without general display. In some of them the buccal were absent. All of them had palm and sole lesions and only four of them had buttock lesions. Everybody recovered after a short eruptive period and without any complications.


[Epidermotropic reticulosis (Woringer-Kolopp disease)].

The Woringer-Kolopp's disease must be considered a clinical-pathologic variant of mycosis fungoides. It is characterized by heterogenous clinical features and a peculiar histological aspect given by the cellular infiltration of the epidermis. The orgin of the reticular elements is discussed. Though some investigators think they originate in the dermis, others consider their origin to be "in situ". The evolution and prognosis are not well defined, as is the case with the hyperplastic, heteromorphic and polymorphic reticuloses. The case presented has had Hodgkin's disease and then developed a picture of pagetoid reticulosis.


[Esophageal localizations of Darier's disease. Radiological description apropos of 4 cases].

The oesophagic affectation seems very frequent in Darier's disease. The authors have found oesophagic lesions in 4 cases examined. The radiologic aspect of this localization shows an oesophagic border in "post stamp" and a definite irregularity of the fasciculation. This fibroscopic examination shows the presence of keratotic papules similar to the skin lesions of this disease.


[Uncombable hair syndrome].

Two cases of new hair defect syndrome, recently described by French dermatologists, are reported. They are characterized by clinically dry, bright and uncombable hair. Pathologic and scanning electron microscope studies, show: -- Angulation of the hair bulb with adherence between the inner root sheath and the hair shaft. -- Canalicular formations longitudinally disposed on the cuticular surface with irregular stratification of the external sheath. A malformation of the hair bulb of unknown aetiology is postulated. This malformation, probably familial, is not associated with any other abnormalities. This defect would result in hair shaft surface abnormality.


[Cowden's disease].

We report a typical case of Cowden disease or Multiple Hamartoma Syndrome in a 32 year old female without a family history of this illness. The patient suffered bilateral fibrocystic disease of the breast, having been subjected to mastectomy because of carcinomatous degeneration. She complained of gastric symptoms, and the x-ray examination showed multiple polyposis. On physical examination an asymptomatic thyroid nodule was noted. The skin showed multiple wart-like lesions, lichenoid papules, localized hyperkeratosis and pigmented spots. She had many papillomas in her mouth a rasberry tongue and bilateral perleches.


[EMG syndrome. Review apropo of 2 cases].

Two cases of EMG syndrome are reported (exomphalos, macroglossia, gigantism). One of them died some hours after the birth. A review of clinical findings, pathogenesis and evolution is presented. Only 60 cases of this rare disease are reported in the world literature.


[A case of lichen nitidus].

A case of Lichen Nitidus exclusively localized on the arms and palms of an 8 year old girl with ten years of evolution and a good response to treatment, is described.


[Multiple trichoepithelioma with variable evolution of its lesions (malignancy and spontaneous remission)].

A case of Multiple Trichoepithelioma with extensive skin involvement without familial history is reported. Some of the lesions showed spontaneous remission over the years, and in two, the microscopcal examination, demonstrated malignant changes (Basal cell Epithelioma).


Central auditory imperception.

The development of clinically applicable techniques for the evaluation of hearing impairment caused by lesions of the central auditory pathways has increased clinical interest in the anatomy and physiology of these pathways. A conceptualization of present understanding of the anatomy and physiology of the central auditory pathways is presented. Clinical tests based on reduction of redundancy of the speech message, degradation of speech and binaural interations are presented. Specifically performance-intensity functions, filtered speech tests, competing message tests and time-compressed speech tests are presented with the emphasis on our experience with time-compressed speech tests. With proper use of these tests not only can central auditory impairments by detected, but brain stem lesions can be distinguished from cortical lesions.


Suitability of antidigoxin antiserum for digoxin in plant extracts.

Antiserum specificity for digoxin was demonstrated by means of three tests: cross-reactivities test, dilutions test and thin-layer chromatography test. The cross-reactivities test examined quantitatively the cross-reactivities with specific compounds and provided an explanation for the basis of the dilutions test. The dilutions test was used to demonstrate the suitability or unsuitability of the antiserum for use in the radioimmunoassay of digoxin in a crude biological sample. The tlc test examined for antiserum cross-reactivity with digoxin and related compounds in a crude extract.


Heart valve replacement by porcine aortic valve (Hancock prosthesis).

Glutaraldehyde preservation of heterograft valves has increased their durability. Sixty-nine Hancock prostheses have been implanted, 30 in the mitral position and 39 in the aortic position. Anticoagulants have not been administered. There has been one important thromboembolic complication and periprosthetic mitral incompetence has occurred in three instances, but prosthetic incompetence has not developed. Two asymptomatic aortic diastolic murmurs have been noted. After a follow-up ranging between two and 19 months, it appears that the Hancock prosthesis offers a valuable alternative to mechanical valve replacement and anticoagulants are not required as a routine.


[Several parameters of the growth of chemostatic culture of Candida utilis in the presence of optimal and submaximal temperatures].

Candida utilis BKM Y-1668 was cultivated in the chemostat (limitation with glycerol) with the rate of flow D from 0.05 to 0.3 hr-1; the economic coefficient Y and Ks were constant at the optimum temperature of growth (30 degrees C). The maximum growth rate was 0.35 hr-1. The content of ATP in the cells and the energy charge of the cell decreased, and the content of ADP and AMP and the activity of phoshohydrolases increased in the cell, with an increase in D from 0.05 to 0.3 hr-1. Small amounts of glycerol and phosphorus were expended for maintaining life without multiplication (m) at 30 degrees C. At the submaximum temperature (40 degrees C), growth of the cells was inhibited, the rate of assimilation of glycerol and phosphorus, and m, increased. The content of ATP in the cells and their energy charge also increased.


[Common action of limiting and inhibiting factors on the growth of yeasts in a continuous culture].

Candida utilis was cultivated under conditions of chemostat and continuous growth which was limited by phosphates. An excess of glucose inhibited the growth. The limiting and inhibiting factors produced simultaneous cooperative action. At a constant velocity of the medium flow, the concentration of biomass decreased, as a result of the inhibition of growth with residual glucose (0.96 and 1.36 per cent), by 5.5 and 15.6 per cent of the absolute value, respectively. The economic coefficient decreased in this case from 40 to 28 per cent. The inhibition of yeast growth resulted also in a decrease of phosphorus content in the cells; moreover, the cells could not utilize all oxygen dissolved in the medium.


[Regulation of synthesis of GTP-cyclohydrolase participating in yeast falvinogenesis by iron].

Pichia guilliermondii, Schwanniomyces occidentalis, Torulopsis candida and several riboflavin-dependent mutants of Torulopsis candida were grown in a medium with a low concentration of iron. In these conditions, the activity of GTP-cyclohydrolase which catalyzes the first step of flavinogenesis increases. The activity of the enzyme increases also when the cells of T. candida and P. guilliermondii with a high content of iron are incubated with alpha, alpha'-dipyridyl which induces overproduction of riboflavin; this action of alpha, alpha'-dipyridyl is eliminated by cycloheximide. Therefore, iron deficiency in the cells of these yeasts causes derepression of GTP-cyclohydrolase participating in riboflavin biosynthesis. The activity of the enzyme is inhibited by FAD but not by FMN and riboflavin.


Redistribution of polycations bound to lymphocytes.

The redistribution of surface-bound polycations on human lymphocytes and mouse-spleen lymphocytes was studied by fluorescence microscopy. A redistribution pattern after incubation with protamine, polylysine and DEAE-dextran resembling patching and capping processes was observed both at 20 degrees C and at 4 degrees C. The interaction between polycations and the cell surface is considered to represent non-specific binding to the membrane proteins. The redistribution process is regarded as precipitaion processes. This is in accordance with the finding that no effect on the binding and redistribution of polycations was noticed in the presence of NaN3, vinblastine sulphate, vincristine sulphate, colchichine, cytochalasin B or trypsin treatment. When incubation periods longer than 5 min. were employed beginning internalization of flourescent material is observed. Interaction with the nuclear membrane similarly resulted in rearrangements and capping.