New antibiotics from the fungus Epicoccum nigrum. II. Epicorazine A: structure elucidation and absolute configuration.

An antibiotic, epicorazine A, isolated from a particular strain of the fungus Epicoccum nigrum, was shown to be a new epidithiodiketopiperazine from its UV, IR, mass, NMR and CD spectra. X-Ray diffraction measures confirmed this structure and its absolute configuration.


New antibiotics from the fungus Epicoccum nigrum. III. Epicorazine B: structure elucidation and absolute configuration.

Comparison of UV, IR, PMR and CD spectra of epicorazine B with those of epicorazine A, a previously isolated metabolite of Epicoccum nigrum, showed that they were isomers with the same epidithiodiketopiperazine skeleton. X-Ray determination of epicorazine B indicated that the difference is related to a cis-trans configuration.


Rapid screening method for alkaline phosphatase activity in cheese: collaborative study.

The method developed for developed for determining alkaline phosphatase activity in cheese, in which phenolphthalein monophosphate is used as the substrate, was collaboratively studied. A 7.5% butanol extract of cheese is reacted with phenolphthalein monophosphate; phenolphthalein is released and yields a red solution that is compared visually with a standard (s) prepared from the same extract. Seven collaborators analyzed 8 samples of cheese, in duplicate, by the screening method and Scharer I method. Of the 208 observations returned, only 4 were incorrect. The alkaline phosphatase method has been adopted as official first action.


Quantitation of added water in milk by using vapor pressure osmometry.

A vapor pressure osmometer (VPO) was successfully studied in 1975. An improved osmometer with greater range and temperature stability was evaluated. Repeatability on 290 mOs/kg standard solution was 289.5 +/- 1.08. The mean and standard deviation on 36 water-free milk samples was 280.5 +/- 4 compared to 280.1 +/- 3 in the previous study. Instrument data from 2 osmometers and 2 cryoscopes produced acceptable chi-square values, and the correlation coefficients between methods were better than 0.99 for samples containing 1--21% added water. In a survey involving 760 samples,  more than 37% of producer samples in one cheese plant contained greater than 3% added water. Additional evidence of added water in market milk samples suggests the immediate application of the VPO to help prevent milk adulteration. The vapor pressure osomometer method for quantitating added water in milk has been adopted as official first action.


Endogenous estradiol-17beta in bovine tissues.

The development of an assay for the estrogen estradiol-17beta in bovine muscle, kidney, liver, and uterine endometrium is described in this report. The tissue was homogenized, extracted, partitioned, and subjected to routine radioimmunoassay. Chromatography was not used. The first part of the report presents data on the accuracy, reproducibility, and sensitivity of the assay. The concentration of estradiol-17beta in the tissues compared with muscle was ranked as follows: uterine endometrium (10-fold higher), liver (3-fold), and kidney (3-fold). The estrogen concentration was not significantly different in the muscle between heifers, but was significantly greater (P less than 0.05) in the liver and kidney of heifers and steers. In heifers pre- and post-estrous phase endometrium (a target tissue of E2beta) contained significantly greater concentrations of estradiol-17beta than did luteal phase endometrium (P less than 0.05), but mean levels in the edible tissues were not significantly different (P greater than 0.05) between these 2 of the 4 stages of the estrous cycle.


Inactivation of the chemoattractant folic acid by cellular slime molds and identification of the reaction product.

Enzymes inactivating the chemoattractants folic acid and pterin were detected in extracellular, intracellular, and particulate fractions obtained from Dictyostelium discoideum strains NC4 and AX-2 and Polysphondylium violaceum. The products of the inactivation reaction were analyzed by thin-layer chromatography and UV spectroscopy. Results obtained indicate that folic acid and pterin were deaminated to 2-deamino folic acid and lumazine, respectively.


Detection of histones by DNA binding ability after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate.

Histones can be detected on the basis of their binding to DNA after electrophoresis in sodium dodecyl sulfate-polyacrylamide gels containing DNA. The method depends on the ability of individual histone components to 1) renature, 2) bind to DNA, and 3) prevent ethidium bromide binding and fluorescence. This technique can provide qualitative and, possibly, quantitative information on histones in crude extracts.


Separate amino and carboxyl procollagen peptidases in chick embryo tendon.

Procollagen synthesized by freshly excised chick enbryo leg tendons is efficiently processed by proteolytic removal of first the amino propeptides and then the carboxyl propeptides. The same processes proceed in confluent short term cell cultures derived from such tendon explants; in sparse cultures cleavage of the amino propeptides predominates. Separate amino and carboxyl procollagen peptidase activities were demonstrated by specific assays in enzymes obtained from cell culture media by ammonium sulfate precipitation, ion exchange chromatography, and velocity sedimentation. Both enzymes are inhibited by EDTA and 1:10 phenanthroline but not by inhibitors of serine proteases. Evidence is provided that the proteolytic scissions are specific and similar to the physiologically occurring processes. The collagen telopeptides left after cutting by the enzymes can participate in lysyl oxidase-induced cross-linking. The enzymes can remove propeptides from cross-linked procollagens without destroying these links which occur through telopeptides. The enzymes act on the separated amino and carboxyl portions of procollagen fragmented by vertebrate collagenase and can act on procollagens which have been associated as well as on molecules in solution.


Substitution of nucleoside triphosphates for ascorbate in the thymine 7-hydroxylase reaction of Rhodotorula glutinis.

Nucleoside di- and triphosphates substituted for ascorbate in the thymine 7-hydroxylase reaction in studies carried out with purified preparations from Rhodotorula glutinis. The stimulations brought about by ascorbate and ATP were found not to be additive. Studies with analogues of ATP indicated that hydrolysis may not need to occur in order for the nucleotide effect to be expressed. The stoichiometry of the production of 5-hydroxymethyluracil and CO2 was not changed by the substitution of ATP for ascorbate. The 7-hydroxylase was found to have considerable thermal stability, and inactivation at 98 degrees C resulted in a parallel loss of the activities effected by ascorbate and ATP. This and the retention of the nucleotide effect upon purification suggest the effect is not mediated through another protein co-purified with the 7-hydroxylase.


Adhesion among neural cells of the chick embryo. IV. Role of the cell surface molecule CAM in the formation of neurite bundles in cultures of spinal ganglia.

The cell adhesion molecule (CAM) is involved in adhesion among embryonic retinal and brain cells and has been detected in a variety of neural tissues. This paper describes the use of spinal ganglion cultures and specific anti-CAM antibodies to determine the distribution of CAM on plasma membranes of nerve processes, and to assess the results of perturbation of its function during the growth of neurites from ganglia. The results indicate that CAM is distributed over the entire surface of nerve processes, and that specific anti-CAM Fab' fragments alter the morphology of neurite outgrowth. In particular, it was observed that anti-CAM inhibits formation of nerve bundles, so that the ganglion becomes surrounded by a tangled net of fine processes. Growth cone functions, such as neurite elongation, motility, and attachment to the substratum, did not appear to be affected by the antibody. These studies suggest that one of the major functions of CAM is to mediate side-to-side adhesion between neurites to form fascicles, and raise the possibility that this molecule serves a key role in embryogenesis of nerve tissues.


Adhesion among neural cells of the chick embryo. III. Relationship of the surface molecule CAM to cell adhesion and the development of histotypic patterns.

We have previously identified a molecule (named cell adhesion molecule [CAM]) that is involved in the in vitro aggregation of neural cells from chick embryos. In the present report, specific anti-CAM antibodies have been used to demonstrated that CAM is localized in neural tissues, and is associated with the plasma membrane of retinal cells and neurites. Furthermore, it has been shown by antibody absorption techniques that the decreased adhesiveness of cultured retinal cells obtained originally from older embryos is correlated with a decrease in the density or accessibility of cell adhesion molecules on the surface of these cells. The central role of CAM in neural cell aggregation has been established by the observation that anti-CAM Fab' fragments inhibit adhesion between neural cells in a variety of assays. To investigate the function of CAM and cell adhesion in developing tissues, aggregates of retinal cells that are capable of forming histotypic patterns in vitro were cultured in the presence and absence of anti-CAM Fab'. The Fab' was found to inhibit sorting out of cell bodies and neurites and to decrease the number of membrane-membrane contacts, suggesting that CAM is associated with cell-cell, cell-neurite, and neurite-neurite interactions.


Localized mast cell degranulation induced by concanavalin A-sepharose beads. Implications for the Ca2+ hypothesis of stimulus-secretion coupling.

Concanavalin A (Con A) covalently linked to Sepharose 4B beads induced localized degranulation of sensitized rat peritoneal mast cells in regions of contact between beads and cells. This degranulation was Ca2+ dependent and was not seen when sensitized mast cells bound to beads conjugated with a nonstimulating lectin, wheat germ agglutinin, or when unsensitized mast cells bound to Con A-Sepharose. The finding that sensitized mast cells which had adhered to Con A-Sepharose beads degranulated in regions of the cell away from the area of bead contact if exposed to soluble Con A excluded the possibility that the localized release was due to a redistribution of the IgE receptors or putative Ca2+ channels to the region of bead contact. The results suggest that, if an influx of Ca2+ is the mechanism for initiating mast cell degranulation, then the opening of Ca2+ channels in the plasma membrane of activated mast cells is a localized event and that Ca2+ acts locally within the cell to initiate exocytosis.


Structure and biochemical composition of desmosomes and tonofilaments isolated from calf muzzle epidermis.

Complexes of plasma membrane segments with desmosomes and attached tonofilaments were separated from the stratum spinosum cells of calf muzzle by means of moderately alkaline buffers of low ionic strength and mechanical homogenization. These structures were further fractionated by the use of various treatments including sonication, sucrose gradient centrifugation, and extraction with buffers containing high concentrations of salt, urea, citric acid, or detergents. Subfractions enriched in desmosome-tonofilament-complexes and tonofilament fragments were studied in detail. The desmosome structures such as the midline, the trilaminar membrane profile, and the desmosomal plaque appeared well preserved and were notably resistant to the various treatments employed. Fractions containing desmosome-tonofilament complexes were invariably dominated by the nonmembranous proteins of the tonofilaments which appeared as five major polypeptide bands (apparent molecular weights: 48,000; 51,000; 58,000; 60,000; 68,000) present in molar ratios of approx. 2:1:1:2:2. Four of these polypeptide bands showed electrophoretic mobilities similar to those of prekeratin polypeptides from bovine hoof. However, the largest polypeptide (68,000 mol wt) migrated significantly less in polyacrylamide gels than the largest component of the hoof prekeratin (approximately 63,000 mol wt). In addition, a series of minor bands, including carbohydrate-containing proteins, were identified and concluded to represent constituents of the desmosomal membrane. The analysis of protein-bound carbohydrates (total 270 microgram/mg phospholipid in desmosome-enriched subfractions) showed the presence of relatively high amounts of glucosamine, mannose, galactose, and sialic acids. These data as well as the lipid composition (e.g., high ratio of cholesterol to phospholipids, relatively high contents of sphingomyelin and gangliosides, and fatty acid pattern) indicate that the desmosomal membrane is complex in protein and lipid composition and has a typical plasma membrane character. The similarity of the desmosome-associated tonofilaments to prekeratin filaments and other forms of intermediate-sized filaments is discussed.


The in vitro assembly of flagellar outer doublet tubulin.

Flagellar outer doublet microtubules were solubilized by use of sonication, and the tubulin was reassembled in vitro into single microtubules containing 14 and 15 protofilaments. The tubulin assembly was dependent on both the KCl and tubulin concentrations, exhibiting a critical concentration of 0.72 mg/ml at optimum solvent conditions. Flagellar tubulin was purified by cycles of temperature-dependent assembly-disassembly and molecular sieve chromatography, and characterized by two-dimensional gel electrophoresis. Although doublet microtubules were not formed in vitro, outer doublet tubulin assembled onto intact A- and B-subfibers of outer doublet microtubules and basal bodies of Chlamydomonas; the rate of assembly from the distal ends of these structures was greater than that from the proximal ends. Microtubule-associated proteins (MAPs) from mammalian brain stimulated outer doublet tubulin assembly, decorating the microtubules with fine filamentous projections.


Electrical coupling and uncoupling of exocrine acinar cells.

The electrical communication network in the mouse pancreatic acinar tissue has been investigated using simultaneous intracellular recording with two separate microelectrodes and direct microscopical control of the localizations of the microelectrode tips. All cells within one acinus were electrically coupled, and the coupling coefficient (the electrotonic potential change in a cell neighboring to the cell into which current is injected [V2] divided by the electrotonic potential change in the cell of current injection [V1]) between two cells near each other (less than 50 micron) was always close to 1. Cells farther apart (50-100 micron) were, in some cases, coupled; in other cases, there was no coupling at all. Coupling coefficients varied between 0 and 1. There was rarely electrical coupling over distances of more than 110 micron. Using microiontophoretic acetylcholine (ACh) application, it was possible to evoke almost complete electrical uncoupling of two previously coupled pancreatic or lacrimal acinar cells from different acini or within one acinus. The effects were fully and quickly reversible. While the ACh-evoked uncoupling in the pancreas was associated with membrane depolarization, ACh caused hyperpolarization in the lacrimal acinar cells. The uncoupling was associated with a very marked reduction in electrical time constant, indicating a reduction in input capacitance (effective surface cell membrane area). The concentrations of stimulants needed to evoke reduction in pancreatic cell-to-cell coupling were 1 micron for ACh, 0.14 nM for caerulein, and 3 nM for bombesin. These concentrations are smaller than those required to evoke maximal enzyme secretion.


Nascent chicken ovalbumin contains the functional equivalent of a signal sequence.

Highly purified mRNA for chicken ovalbumin has been translated in a cell-free protein synthesizing system from rabbit reticulocytes in the presence or absence of EDTA-stripped microsomal membranes from dog pancreas. Nascent--but not completed--ovalbumin was transferred across the microsomal membrane, as demonstrated by cotranslational core glycosylation of ovalbumin nascent chains, by resistance to posttranslational proteolysis of only the glycosylated ovalbumin chains, and by cosedimentation with the membrane of exclusively the glycosylated form. Furthermore, nascent chains of bovine prolactin were observed to compete with nascent ovalbumin for transfer across the microsomal membrane. However, no competition for membrane sites was observed between nascent chains of rabbit globin and either nascent ovalbumin or prolactin. We interpret these results to suggest that nascent ovalbumin contains the functional equivalent of a signal sequence for transfer across membranes, and that membrane components involved in the segregation of secretory proteins with cleaved signal sequences also function in the segregation of ovalbumin.


Cyclic production of tension force in the plasmodial strand of Physarum polycephalum and its relation to microfilament morphology.

Cyclic contraction and relaxation of plasmodial strands of Physarum polycephalum were measured under both isotonic and isometric conditions, and their relation to changes in microfilament (MF) morphology was investigated. The contraction-relaxation rhythm of a strand segment was insignificant and irregular immediately after isolation from the mother plasmodium. It became regular half an hour later when local minute rhythms were synchronized spontaneously. If a strand kept under isotonic conditions was loaded with a heavier weight or a strand kept under isometric conditions was stretched a few times, the amplitude of each contraction wave was enhanced. After a strand had been thus conditioned, it was fixed at a selected phase of the contraction-relaxation cycle under both isotonic and isometric conditions. The state of MFs changed strikingly according to the phase of the contraction cycle. In the shortening phase of the strand under isotonic contractions, MFs with a diameter of 6--7 nm were arranged parallel to each other to form large compact bundles in which adjacent filaments were bridged with cross linkages. Among these MFs, thicker filaments were sporadically scattered. At about the phase of minimal strand length, most of the MFs became kinky and formed networks. In the elongating phase, new loose bundles of MFs developed from the network. These loose bundles became compact again when the strand reached its maximal elongation phase. In the isometric contraction, MFs in the increasing tension phase were nearly the same as those in the shortening phase in isotonic contraction. Around the maximal tension phase, dense areas of MFs appeared along the bundles in place of the network formed in the isotonic contraction phase. These areas were closely packed, with MFs arranged parallel to each other. In the decreasing and minimal tension phases in isometric contraction, MFs were arranged similarly to those in the elongating and maximally elongated phases, respectively, in isotonic contraction. Alternation between the straight bundle and fine network configuration of the MFs observed in isotonic contraction was inconspicuous in isometric contraction. This was probably due to spatial restriction of shortening under isometric contraction. The results are interpreted in terms of cyclic changes of the aggregation pattern of the MFs in the form of F-actin, as opposed to the possibility that the contraction-relaxation cycles depend on cyclic G-F transformation of actin.


Divalent cation stimulation of in vitro fibre assembly from epidermal keratin protein.

Keratin was extracted from purified cornified cells of newborn rats in Tris-HCl-buffered 8 M urea containing beta-mercaptoethanol. Microfilaments were assembled in vitro by reducing the ionic strength of buffer and the urea concentration. One millimolar concentration of KCl and NaCl did not affect filament formation, but the same concentration of divalent cations greatly altered this process. CaCl2 and MgCl2 induced gelation of keratin by formation of bundles of birefringent macrofilaments. ZnCl2, CuSO4 and HgCl2 formed greater numbers of macrofilaments and the protein aggregated.


Recirculation: a uremic syndrome complicating the use of prosthetic arteriovenous fistulas for hemodialysis.

Malfunction of prosthetic arteriovenous fistulas is described in two patients, unheralded by abnormalities of arterial flow or venous resistance. In each case, significant graft recirculation due to stenosis of the venous limb caused the insidious onset of uremic symptoms in patients apparently receiving adequate hemodialysis. The uremic syndromes disappeared after correction of the faulty vascular access. Recirculation may be a "silent" cause of prosthetic graft malfunction which is promptly diagnosed by a simple mathematical formula. Routine estimates by a simple mathematical formula. Routine estimates of recirculation should be performed in any chronic hemodialysis patient with relapse of uremic symptoms or suspicious serum chemistries despite regular, uncomplicated hemodialysis. Patients found to have significant recirculation should be considered for angiography and graft replacement or revision.


Computer analysis of cervical cells. Automatic feature extraction and classification.

A prescreening instrument for cervical smears using computerized image processing and pattern recognition techniques requires that single cells in the specimen can be automatically isolated and analyzed. This paper describes a dual wavelength method for automatic isolation of the cytoplasm and nuclei of cells. Density-oriented, shape-oriented and texture-oriented parameters were calculated and evaluated for more than 600 cells. It is shown that the computer can be taught to distinguish between normal and atypical cells with an accuracy of ca. 97%, while human classification reproducibility is ca. 95%. In addition, an attempt to assign a measure of atypia to individual cells is described.


Effect of ionizing radiation of the antigenic composition of typhoid bacteria.

Changes in the antigenic composition of typhoid bacteria occurring during the exposure of microbial suspension to different doses of gamma irradiation [Co60] ranging between 0.5 and 3.0 Mrad were studied. Immunoelectrophoresis in agar was used to determine the antigenic composition of different samples of irradiated bacteria. The antigenic composition of bacteria irradiated with doses up to 2.5 Mrad was found to be similar to that of non-irradiated bacteria. Antigens demonstrated by means of Vi, H and O ontisera are preserved in these bacteria. However, all irradiated bacteria in general slightly differ from non-irradiated bacteria; this is manifest in a different configuration and position of the precipitation lines in the cathodic part of the immunophoreograms. The content of the component migrating rapidly towards the cathode, evidently the O antigen in the R form, in the irradiated bacteria increases with the dose of radiation. No new serologically active substances, non-existent in non-irradiated bacteria, were found to appear in the process of irradiation.


Investigation of common and specific components of Cl. septicum and Cl. histolyticum toxins.

Interrelations between the common and specific components in the toxins of several strains of Cl. septicum and Cl. histolyticum were investigated. The method of tissue culture, which yields more stable results than biological tests on animals, was used. It has been demonstrated that native toxins of Cl. seticum (7 strains) and Cl. histolyticum (7 strains) cause cytotoxic changes in chick embryo fibroblasts. These changes are similar to each other and identical with changes occurring under the effect of concentrated toxins of the mentioned microorganisms. Cross reactions of neutralization with antitoxic and species-specific sera against Cl. septicum and Cl. histolyticum have shown that the strains of Cl. septicum and Cl. histolyticum synthesize toxins with components possessing common antigenic properties. The strains of Cl. histolyticum synthesize a greater amount of components common with Cl. septicum than the strains of Cl. septicum in which the amount of heterologous antigens varies.


Experimental investigation of changes in the antigenic structure of influenza viruses during their persistence.

Systems of persistent influenza infection produced in the cultures of human embryonal kidneys and in the diploid cells of human embryonal lungs under the effect of A/Hong-Kong/1/68 and A/Victoria/35/72 viruses were characterized. The cell systems were studied in the course of 4--3u passages, i. e., from 40 to 289 days. A total of 102 viruses were isolated during the period of observation, out of which 44 were examined for the antigenic profile of HA and NA. The antigenic structure of the initial viruses was preserved in 31 variations isolated while in 13 variants it was pronouncedly and permanently changed. The following mechanisms of the mentioned phenomenon are discussed: mutations, intrapopulation recombinations and integrations.


Changes in lactate and malate dehydrogenase isoenzymes in salmonella under the effect of potassium dichloroisocyanurate.

The method of enzyme-electrophoresis in agar gel according to Wieme (1959) was used for the study of lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) isoenzymes of 24-hour and 48-hour Salmonella cultures exposed to a 0.02% solution of potassium dichloroisocyanurate (PDIC). Severe repression of LDH and MDH isoenzymes was observed immediately after the exposure of the culture to the disinfectant solution. A significant decrease in the content of the isoenzyme LDH1 and of the cytoplasmic fraction (C1) of MDH simultaneously with the appearance of the fractions LDH4, LDH1a and LDH1b were established in the strains cultured on MPA in the course of 24 hours following the exposure. A tendency to a decrease in the LDH1 content was preserved in the experimental cultures after 48 hours, but the spectrum of MDH isoenzymes showed almost no differences in comparison with that of MDH isoenzymes in 48-hour cultures of the control strains.


Studies of local reactivity to B. pertussis suspensions by means of the rat paw oedema.

In the present study, the authors verified earlier experience with the rat-paw-oedema test and tried to improve the method and to increase its sensitivity so as to make possible, by means of this test, to screen vaccines producing different reactivity. Toxic and non-toxic B. pertussis suspensions were used as test material. The sensitivity of the animal to the suspension was increased by sensitization of the rats using Alditepera and by influencing the pituitary using different doses of Hydrocortisone. Influencing the pituitary by means of cortisone was found to be more suitable for screening assessment of the reactivity to suspension than sensitizing the animals.


Allergy to insect stings. IV. Diagnosis by radioallergosorbent test (R.A.S.T.).

Radioallergosorbent tests (RAST(s)) have been developed and assessed for the diagnosis of insect hypersensitivity by using a purified allergen from honeybee venom, phospholipase A, and crude yellow jacket venom. Sera from 193 patients positive both by history and skin test to one of these insects were compared with various groups of control sera. Eighty percent of sera from skin test-positive patients were RAST positive; positive RAST were found in 16% of sera tested from skin test-negative patients. A highly positive RAST correlates well with a positive skin test and clinical sensitivity, but serum IgE is not measurable in many patients with mast cell or basophil bound antibody. Since biologically important reactions of antigen with IgE require that the antibody be cell bound, skin testing would be preferred to RAST if one were limited to a single test for the diagnosis of insect allergy.


Complement lysis: evidence for an amphiphilic nature of the terminal membrane C5b-9 complex of human complement.

The terminal, membrane-derived C5b-9 complex of human complement (C) is an apparently hollow, cylindrical macromolecule vertically oriented on the target membrane. In the present study, an antiserum to the complex has been used to probe its immunobiochemical properties. "Neoantigenic" determinants characteristic of the complex have been detected, which are absent on native C5-C9 molecules. Evidence that the C5b-9 complex is an amphiphilic molecule that possesses apolar, detergent-binding surfaces has been obtained by using charge-shift crossed immunoelectrophoresis, and by direct demonstration of Triton X-100 binding to the complex in quantitative immunoelectrophoresis. By the same criteria, serum C5, C6, and C9 are hydrophilic molecules. The results indicate that assembly of C5-C9 into the terminal membrane C5b-9 complex is accompanied by conformational changes in the individual C components that lead to the exposure of apolar molecular regions in the complex. It is proposed that this constitutes the basis for the lipid-binding properties of the macromolecule, which enable it to become inserted into biologic and artificial lipid membranes with apparent generation of a transmembrane pore.


Detection of specific antibody producing cells in porcine colostrum by in ovo translation of their mRNA.

An improved method is described for the determination of antibody producing cells in sows colostrum. The test system comprises in ovo translation of mRNA from swine colostral cells and analysis of the translation products by radioimmunoassay with specific antibodies and antigen.


A scanning electron microscopic study of the neural crest cell cytoskeleton after triton x-100 demebranation.

Triton x-100, which solubilizes cell membranes, was used to expose the detergent resistant cytoskeletal systems of cultured day 4 (angulated) and a day 8 (STELLATE) NEURAL CREST cells. This procedure reveals a network of fibers which correlates structurally with the morphological state of neural crest cell differentiation.


DNA sequences in influenza virions.

During the propagation of A (H3N2) influenza virus in chick embryos, incorporation of 3H-thymidine into virions takes place, whereas no such incorporation occurs with Newcastle disease virus. Incorporation of 3H-thymidine is a result of DNA synthesis. This virion-associated DNA is present in cores obtained after treatment of virions with bromelain.


Effect of interferon on the cell cycle of BALB/c 3T3 cells.

The effect of interferon on the exponential growth phase of BALB/c 3T3 cells was studied. Although interferon reduced the growth rate and the proportion of cells in both the M and S phases (mitotic index and labelling index), there were no appreciable differences in the duration of these phases between control and interferon-treated cells. Moreover, the shape of the first peak of the fraction of labelled mitoses (FLM) curve was not altered by treatment with interferon, which indicates that the duration of the S and G2 phases was not affected. However, the height of the second peak of the curve in interferon-treated cells was extremely reduced as compared to control culture. These results are compatible with the idea that the suppressive effect of interferon is exerted mainly in the G1 phase (A-state) of the cell cycle of BALB/c 3T3 cells.


Ultrastructural study of virus-like particles in Chinese hamster lung cells.

Virus-like particles were found in the E36 cells. One type, associated with centrioles, consisted of two concentric shells with a diam. of 50 to 60 nm. Some of these particles were seen budding through the plasma membrane giving rise to a free immature particle showing two concentric shells and an outer envelope. Concentration of the inner shell into a nucleoid results in a mature particle characterized by a nucleoid, mainly eccentrically located, surrounded by an intermediate layer and wrapped in an envelope. The diameter of the nature and immature particles was 75 to 85 nm. The morphogenesis of this virus-like particle resembles that of the oncoviruses.


Characterization of adenovirus type 12 tumour antigen produced in chick fibroblasts.

Tumor (T) antigen was characterized in non-permissive chick fibroblasts and permissive HEp-2 cells infected with Ad12 either in the presence or in the absence of cytosine arabinoside. Antiserum against T antigen specifically immunoprecipitated two polypeptides of apparent mol. wt. 50 000 and 11 000.


The mutagenic assay of some hair dye components, using the thymidine kinase locus of L5178Y mouse lymphoma cells.

Five hair dye components were tested for their mutagenicity at the TK +/- locus of L5178Y cells. Three of the components, m-phenylenediamine, 2 nitro-p-phenylenediamine, and 4-nitro-o-phenylenediamine, gave a positive response that was dose related; 2,4-diaminoanisole gave a questionable response; and 2,5-diaminoanisole gave a negative response. The assay was carried out in vitro without metabolic activation during a 24-hr chemical exposure period. It is concluded that further in vivo tests are necessary to establish the safety of the hair dyes.


Presence of a covalently linked protein on calicivirus RNA.

The infective RNA of the calicivirus, vesicular exanthema virus, has been shown to contain a protein which is apparently linked to the RNA by a covalent bond. The protein remained bound to the RNA after boiling with SDS-mercaptoethanol-urea or treating with formamide-dimethylsulphoxide but was removed by incubating with proteinase K. The mol. wt. of the protein was estimated to be about 1o X 1O(3) by electrophoresis in highly cross-linked polyacrylamide gels. The infectivity of the RNA was destroyed by removal of the protein with proteinase K.


Correlation between actin polymerization and surface receptor segregation in neuroblastoma cells treated with concanavalin A.

In response to concanavalin A (Con A), neuroblastoma cells undergo marked morphological changes which involve the retraction of neurites and the induction of broad and extensive lamellar regions around the cell periphery. From the use of FITC-Con A it was shown that the membrane formed on the induced lamellar regions lacked receptors to Con A from the onset of lamella formation. These receptors were confined to the cell body; they initially showed a uniform distribution and were subsequently collected into patches and finally into aggregates or caps. When the aggregates occurred on the cell periphery their position coincided with areas free of lamellae. Investigations of the lamellar regions in Triton-extracted cell monolayers showed them to consist of a meshwork of actin filaments containing radiating thin filament bundles or microspikes. With increasing time in the presence of Con A there was a progressive increase in the number of radiating microspikes. Previous studies have shown the actin in these lamellar regions to be singly polarized with respect to the cell body. From the segregation of Con A receptors away from areas of actin polymerization in the lamellae it is concluded that actin is involved in some indirect way in surface receptor movement.


Effect of dietary protein on urinary excretion of 3-methylhistidine in rat.

The excretion of the amino acid 3-methylhistidine in urine has been shown to be correlated with protein catabolism in skeletal muscle. In rats, 3-methylhistidine is partly acetylated (N-acetyl-3-methylhistidine) and it has been proposed that the relative amounts of 3-methylhistidine and N-acetyl-3-methylhistidine in urine is age dependent. In this experiment the effect of dietary protein quality on urine excretion of 3-methylhistidine and N-acetyl-3-methylhistidine was studied. Six groups of rats (mean weight 80 g) were fed diets containing 10% protein of different quality, the net protein utilization ranging from 76.7 for egg albumin to 28.9 for wheat gluten. The excretion of non-acetylated 3-methylhistidine was not dependent on the diet. There was, however, a good correlation between protein quality and total urine 3-methylhistidine (3-methylhistidine plus N-acetyl-3-methylhistidine), the higher the protein quality, the greater being the excretion of total 3-methylhistidine. The relative amounts of 3-methylhistidine and N-acetyl-3-methylhistidine correlated with the mean body weight, but not the age, of the animals. This study therefore demonstrates that a relationship exists between the nutritive quality of the dietary protein and the urinary excretion of 3-methylhistidine in rats.


The effect of chronic ethanol ingestion on myocardial glucose and energy metabolism.

To determine the effect of chronic alcohol ingestion, rats were given 15 or 25% v/v of alcohol in water for a period of 6 months. The activities of some key enzymes involved in the metabolism of glucose, mitochondrial respiratory rates, and efficiency of oxidative phosphorylation were studied in the hearts of alcohol-treated and untreated rats. In the group receiving 15% alcohol, glucose-6-phosphate dehydrogenase (G-6-PDH) was elevated. In rats given 25% alcohol, activities of G-6-PDH, aldolase, and glyceraldehyde phosphate dehydrogenase were elevated but isocitrate dehydrogenase was reduced. Mitochondrial respiratory rates and the efficiency of phosphorylation were depressed in rats given 25% of alcohol. Except for mitochondrial oxidation of pyruvate and alpha-ketoglutarate, all biochemical parameters studied were within normal limits a month after alcohol was discontinued.


Duodenal carcinoid with cytoplasmic whorls of microfilaments.

A unique ultrastructural feature in a duodenal carcinoid is described, viz., large juxtanuclear accumulations of microfilaments arranged in whorls. Similar microfilaments are found in the D1 cell of the gastro-entero-pancreatic endocrine system. Apparently identical whorl-like accumulations of filaments have been previously described in bronchial cells of animals treated with nitrosamines and in human giant cell carcinoma of the lung. We suggest that the D1 cell is the cell of origin for the described carcinoid as well as for the above-mentioned tumours of the lung.


Hodgkin's disease: an immunohistochemical and histological study.

The distribution of immunoglobulin (Ig) in Hodgkin's tissue has been demonstrated in paraffin and cryostat sections by the unlabelled antibody peroxidase-antiperoxidase (PAP) method and the two-stage fluorescein isothiocyanate (FITC)-based technique. The morphology and histogenesis of the Reed-Sternberg (RS) and Hodgkin (HD) cells has been studied with the metalophil method of Marshall (1948) and this revealed a population of dendritic histiocytes which corresponded in number, sizes and distribution to the RS and HD cells in adjacent sections stained by haematoxylin and eosin. The largest RS cells, however, appeared to be non-dendritic. A notable feature of Hodgkin's tissue was the tendency for the dendritic cells to form "nodules", in combination with a population of small lymphocytes. The PAP technique reveals Ig in the form of mu and delta heavy chains, as well as kappa and lambda light chains, in close association with and probably on the surface of a high proportion of these lymphocytes, suggesting that they are immature B cells. Similar reactions were given by the mantle of lymphocytes of surviving normal lymphoid follicles, and metalophil dendritic histiocytes were also demonstrated within the mantle and subjacent part of the germinal centre. Numerous immunocytes containing Ig were present in all lesions; in the majority of cases, more cells contained gamma than alpha or mu heavy chains, although in these cases alpha-positive cells outnumbered those containing gamma or mu heavy chains. In two-thirds of the cases, one-third of the RS cells contained cytoplasmic immunoglobulin. This was exclusively gamma heavy, although both light chains were present. In about half the cases a minority of the HD cells contained gamma chain. The results suggest that HD and RS cells are dendritic histiocytes of the type normally found in the lymphoid follicles and that their tendency to form nodules in assoication with B lymphocytes is a manifestatin of this origin. It is suggested that the presence of Ig most probably results from absorption of antigen-antibody complexes on the cell surface.


Amino acid, heme, and sterol requirements of the nematode, Rhabditis maupasi.

Rhabditis maupasi, a nematode that occurs in the mantle cavity of Helix aspersa and related North African food snails, requires 5 amino acids (lysine, methionine, phenylalanine, tryptophane and valine) for maintenance of the stage-3 survival larvae, and 5 additional amino acids (arginine, isoleucine, leucine, threonine and, marginally, histidine) for development of these larvae into adults and for reproduction. Tyrosine is beneficial but not absolutely required for reproduction. These results were obtained with axenic R. maupasi grown in a chemically defined medium containing salts, trace metals, purines and pyrimidines, Krebs cycle intermediates, a fatty acid (butyric), vitamins, urea, and a carbohydrate (dextrose). The complete medium contained 18 amino acids; each of 18 test media was deficient in one of the amino acids. In the complete medium and in a medium lacking the "nematode nonessential" amino acids, stage-3 R. maupasi developed into adults and produced one generation of offspring. For continuous cultivation, however, the nematode also required hemin or another iron porphyrin as well as a sterol such as cholesterol.


Polyamine metabolism III: urinary acetyl polyamines in human cancer.

Polyamine levels were determined by high-pressure liquid chromatography in the unhydrolyzed 24-hr urine obtained from 15 cancer patients and nine normal subjects. N1-Acetylspermine, N4-acetylspermine, N4-acetylspermidine, and N-(3-aminopropyl)acetamide were not detected in any samples. N1-Acetylspermidine, N8-acetylspermidine, acetylputrescine, and acetylcadaverine were present in all samples. Furthermore, acetylspermidine and acetylputrescine were excreted in much greater quantities than the respective amines in the urine of both cancer patients and normal subjects. The levels of N1-acetylspermidine were considerably higher than the levels of N8-acetylspermidine in the 24-hr urine of cancer patients, resulting in a ratio of N1- to N8-acetylspermidine in the urine of cancer patients significantly higher than that in normal subjects. The urinary levels of acetylputrescine were also significantly higher in cancer patients. The urinary polyamines in 13 of the 15 cancer patients were outside the 95% confidence limits of the normal mean for the ratio of N1- to N8-acetylspermidine and acetylputrescine. All cancer patients showed values outside the 95% confidence limits of the mean for either of these two parameters. Diurnal variation was observed in the urinary excretion of the acetyl polyamines but not for the free amines in two normal subjects.


Magnetic microspheres: synthesis of a novel parenteral drug carrier.

The synthesis and characterization of a novel parenteral drug carrier capable of area specific localization by magnetic means are described. The carrier consists of human serum albumin microspheres, average of 1 micrometer in diameter, in which a magnetizable material (magnetite) and a prototype drug (doxorubicin) are entrapped. Stabilization of the microsphere matrix by formaldehyde, 2,3-butanedione, and heat conferred equal stability to the matrix but differentially affected the in vitro drug release rate. Release doxorubicin was chemically identical to the starting material.


Activity of enzymes involved in pyrimidine nucleotide synthesis during uterine growth in cyclic rats.

The pyrimidine nucleotide salvage enzymes, uridine and thymidine kinases, and the enzyme, aspartate carbamyltranferase, involved in de novo synthesis, were investigated to assess their relative contributions to the RNA and DNA precursor requirements of uterine growth in cyclic rats. Only aspartate carbamyltransferase reflected the known fluctuations in plasma oestradiol-17beta and uterine mitotic activity, being minimal at dioestrus 1 and maximal at pro-oestrus. Treatment of ovariectomized rats with oestradiol-17beta stimulated carbamyltransferase activity, but cycloheximide prevented this response, indicating the association between the enzyme and this hormone. Uridine kinase activity did not vary during the oestrous cycle, but there were peaks of thymidine kinase activity on the days of pro-oestrus and the 1st day of dioestrus.


Storage of spermatozoa in the female genital tract of the vespertilionid bat, Scotophilus heathi.

Spermatozoa were observed in all parts of the genital tract in females killed in January, February, March and April although copulation occurs only until early February. The spermatozoa were arranged linearly with their heads orientated towards the epithelial lining.


Factors affecting reproduction in red deer (Cervus elaphus) hinds on Rhum.

In a population of free-ranging red deer hinds on the Isle of Rhum (Inner Hebrides) we investigated relationships between four aspects of reproductive performance (fertility, calf birth weight, birth sex ratio and calving date) and four variables likely to affect the mother's condition: age, reproductive status, home range area and year of calving. Fertility was significantly related to mother's age, reproductive status and home range area as well as to year of calf's birth. Stag calves were heavier than hind calves and birth weight was significantly related to mother's age, home range area and year of (calf's) birth but not to mother's reproductive status. Birth sex ratio did not differ from parity, and was not significantly associated with any of the four variables examined. Birth date was significantly related to the mother's reproductive status, home range area and year of (calf's) birth but not to mother's age or the sex of the calf.


Maternal age and the incidence of aneuploidy in first-cleavage mouse embryos.

Eggs obtained from young virgin and aged parous female mice were fertilized in vitro to compare the incidence of aneuploidy in the resulting first-cleavage embryos. There was a significantly higher incidence of aneuploidy in the aged group (7.5% versus 3.3%) and this was due solely to a higher proportion of trisomic female chromosome complements; there was no difference in the incidence of monosomics. These results thus suggest that non-disjunction occurs more frequently in eggs from older females, leading to the production of aneuploid embryos.


Cephradine and epicillin in body fluids of lactating and pregnant women.

The passage of two antibiotics, cephradine and epicillin, into the milk of 12 lactating women and the amniotic fluid of 48 pregnant women was investigated. All women were given a 500-mg capsule every six hours for at least two days prior to our taking multiple biologic samples from them. Constant levels of both antibiotics were reached in the milk of lactating women as well as in the amniotic fluid of mid-term and full-term pregnant women during the sample period. The ratio of drug concentrations in serum to that in milk was about 5.0 for each antibiotic. In the midtrimester women, the ratio of concentrations in serum to amniotic fluid of both antibiotics was approximately 1.0, suggesting the development of an equilibrium between these two compartments. This ratio was 0.2 for both drugs in the full-term women, demonstrating that the antibiotics concentrated in the amniotic fluid compartment.


Synthesis and pharmacological activity of a phosphate ester of delta8-tetrahydrocannabinol.

A water-soluble phosphate ester of delta8-tetrahydrocannabinol (delta8-THC) was synthesized and its pharmacological activities were examined. The cataleptogenic and thiopental sleep-potentiating effects of delta8-THC phosphate in the mouse was approximately 10 and 7% of those of delta8-THC, respectively. However this phosphate showed almost the same potency and a longer duration of hypothermic effect, as compared with delta8-THC in the mouse. The acute toxicity of this phosphate was far lower than that of delta8-THC. delta 8-THC phosphate was difficultly hydrolyzed by alkaline phosphatase or mouse liver homogenate in vitro. The mode of action of the phosphate derivative is discussed in connection with this enzymatically difficult hydrolysis.


2,5-Bis(3,4-dimethoxybenzyl)cyclopentylamine, a peripheral dopamine blocking agent.

2,5-Bis(3,4-dimethoxybenzyl)cyclopentylamine hydrochloride has been synthesized. The intermediate 2,5-bis-(3,4-dimethoxybenzyl)cyclopentanone was formed in 91.8% yield using a sodium methoxide catalyzed aldol condensation and catalytic reduction. The oxime of this ketone was catalytically hydrogenated to the amine which was converted to the hydrochloride (76%). The amine hydrochloride was found to be an effective antagonist to the low-dose hypotensive effect of dopamine; the half-life of this effect was 18 min. At dopamine doses of 3 mg/kg in the atropinized and phenoxybenzamine treated dog, the ED50 for blockade was 4--5 mumol/kg. In direct contrast to its peripheral dopamine blocking activity, the compound potentiates apomorphine-induced stereotypy.


The jelly envelopes and fertilization of eggs of the newt, Notophthalmus viridescens.

Fertilization in Notophthalmus viridescens is internal and involves passage of the sperm through five layers of egg jelly (J5-J1, from outermost to innermost), each of which is secreted by a discrete region of the oviduct. Polyspermy is normal. Passage of the sperm through the jelly and into the egg was studied by a technique of artificial insemination similar to natural insemination, in that undiluted fluid from the vas deferens was applied directly to eggs with various layers of jelly present, followed by flooding with water three to five minutes later. In general, successful fertilization increased as the number of jelly layers increased; jellyless coelomic eggs were not fertilizable. Sperm passage through the jelly and into the egg usually occurs within one to three minutes. Upon hydration of the jelly, barriers to sperm penetration develop in layers J5 and J3. Changes in the egg jelly thus seem to be involved in the restriction of polyspermy to a low level.


Sexual dysfunction in women who were molested as children: one response pattern and suggestions for treatment.

A common pattern of women who were sexually molested as children and seek treatment for sexual dysfunction is described. The arousal, rather than the orgasmic, component is involved. The reasons for the evolution of this particular dysfunction are discussed. Treatment should extend the period of sensate focus and address the issues of the woman's anger, her need for control, and her guilt. Excellent results can be achieved in helping the patient to experience physical intimacy and the pleasure of sexual arousal.


Interdisciplinary treatment of male sexual dysfunction in a military health care setting.

An interdisciplinary approach to the rapid treatment of male sexual dysfunction is presented. Clinical results for 36 sexually dysfunctional men, ages 20 to 61 are stated. Increased communication between physicians and sex therapists results in improved patient care and increased satisfaction for staff and patients. Interdepartmental cooperation within a health care setting maximizes effective use of personnel and knowledge resources.


Effects of erotic stimulation and masturbatory training upon situational orgasmic dysfunction.

Six single women, aged 22 to 29 years, were treated in a laboratory situation through erotic stimulation with masturbatory training for the disorder of situational orgasmic dysfunction. With single subject designs, three conditions of treatment were counterbalanced to estimate component effects. Intervention conditions included exposure to selected erotic stimuli, self-masturbation, and the preceding simultaneous combination. Frequency of orgasm was monitored via heart rate and verbal confirmation. Erotic stimulation with masturbatory training proved adequate to establish and maintain orgasmic responsiveness. Follow-up measures, conducted 6 to 12 months thereafter, partially supported generalization of treatment effects across environments and into existing heterosexual patterns of behavior.


The treatment of sexual dysfunction in gay male couples.

This paper reports a 2-year experience in treating sexual dysfunction in 22 gay male couples. The full range of arousal, orgasmic and desire phase disorders as well as case examples and variations in treatment techniques are discussed. To date, this comparative neglected area of research and treatment has yielded encouraging results.


[The treatment of urinary retention in the woman by partial urethrectomy (author's transl)].

Urinary retention due to a urological cause is relatively rare in the woman. As in the male, it is associated with dysuria, frequency, bladder distension and renal failure, and nevertheless is often unrecognised. Attributed to bladder neck disease, treatment by endoscopic resection gives few good results. The abnormal length of the urethra (35 to 60 mm) in women with retention led us to shorten the urethra by ablation of the meatus and resection of 10 to 15 mm of the terminal urethra in order to obtain a final length a little shorter than the average, less than 30 mm. This shortening involves opening of the urethrovesical angle and restores micturition. This simple technique, free of mortality, resulted in micturition without residue in 12 of the 15 patients operated upon and seen at long term follow-up. It may be used without difficulty when another technique has failed; endoscopic resection, YV plasty, dilatations.


Mutants of Sindbis virus. III. Host polypeptides present in purified HR and ts103 virus particles.

The amounts of host-encoded protein present in purified Sindbis virions of both the HR strain and of a mutant (ts103) which makes multicored particles were examined. Cells were labeled with [35S]methionine before infection and with [3H]methionine postinfection. Virions were purified by velocity sedimentation and isopycnic banding, and their polypeptides were examined by polyacrylamide gels in a sodium dodecyl sulfate-containing discontinuous buffer system. Host prelabeled material was found principally in a small number of discrete polypeptides in HR virions, which contained as little as 0.2% host-encoded protein. Virus-sized particles of mutant ts103 contained significantly more host material, and multiploid particles from ts103 infection contained up to 12% host prelabeled protein.


Charcot-Marie-Tooth disease and schizophrenia in identical twins.

Charcot-Marie-Tooth disease and schizophrenia occurred in monozygotic twins. Dermatoglyphics and blood grouping determined monozygocity. Charcot-Marie-Tooth disease appeared to be an autosomal dominant trait transmitted from the mother. No definite evidence of inheritance was seen for schizophrenia.


Preference for morphine and drug-seeking behavior in morphine dependent rats.

We have already reported that morphine pretreated rats prefered morphine-admixed food during choice trials with the two cup method. In the present work, we utilized both the five and two cup methods and observed the preference for morphine in rats forcedly pretreated with the drug and the increasing rate of preference for the drug in rats where the feeding time was limited. In morphine pretreated rats, preference rate for morphine was 61.2 +/- 3.0% with the five cup method and 61.8 +/- 3.3% with the two cup method during the choice trials. In rats that were limitedly treated with morphine, each preference rate for morphine during choice trials rapidly increased in the five cup method, i.e. 5.5 leads to 14.4 leads to 31.7 leads to 43.6 leads to 61.2%, and the preference rate for morphine stabilized at approximately 60 percent. Findings with the two cup method were similar. After the preference for morphine was stabilized at the 60 percent level, morphine was given subcutaneously and it was found that the preference rate was dependent on the dose injected. When the drug-admixed concentration was changed from 1 mg/g food to 0.5 and 2 mg/g food, the preference rate changed in parallel with the concentration. When the number of food cups containing morphine-admixed food was changed from 1/5 to 2/5, 3/5 and 4/5 food cup, the preference rate was not effected. These studies clearly demonstrate drug-seeking behavior in rats. In the process of preference for morphine, morphine treatment enhances spontaneous intake of morphine-admixed food.


Control of oxyurids in mice using thiabendazole.

Mice with a natural infection of Syphacia obvelata and Aspiculuris tetraptera were continuously medicated with 0.1% thiabendazole in the diet. No oxyurids were found in test animals after 24 days of treatment though control animals remained infected. Growth rates in treated animals increased. 2 generations of mice were studied and, although overall production fell by 1.45%, the use of this drug in total eradication of the worms is suggested.


Metabolic stability of the nucleoside transport system of Novikoff rat hepatoma cells.

Rates of transport of uridine and thymidine, estimated with a rapid sampling technique, did not change with culture age. Inhibition of cellular RNA and protein synthesis for periods up to 6 h, did not lead to a loss of nucleoside transport activity. Mild treatment of cell suspensions with trypsin or neuraminidase had no effect on the kinetics of thymidine transport. Thus we conclude, contrary to previous reports, that nucleoside transporters are metabolically stable and that the decreases in nucleoside uptake rates observed with decreased protein synthesis reflect loss of nucleoside kinase activities. These kinases (which have narrow substrate specificity) rather than the membrane-associated, transport apparatus (which has broad substrate specificity) are the most likely sites for regulation of nucleoside uptake.


A sensitive solid phase radioimmunoassay for secretory IgA: its application for the determination of urinary levels of secretory IgA in children and adults.

A sensitive solid phase radioimmunoassay method was established for the specific quantitative determination of secretory IgA (sIgA) by taking advantage of the dual antigenicities of sIgA, one specific for alpha-chain and the other for secretory component (SC). The sIgA and IgA in the sample were first bound by anti-IgA antibodies coated on the polystyrene tube, then the amount of bound sIgA was quantified by the use of 125I-labeled anti-SC antibodies. This method is quite sensitive and allows us to distinguish sIgA from IgA and free SC which usually coexist in exocrine secretions. Linear relationship was observed between the bound radioactivity of radioiodinated anti-SC and the amount of sIgA in the range of 5 to 60 ng of sIgA. With this method, the urinary sIgA levels in normal children and adults were measured. Urinary sIgA was detected in half of infants within 7 days after birth, while it could be demonstrated in almost all of the infants by 14th day. Then, it gradually increased and reached about a half of the adult level by the age of puberty. This method will be useful for the sensitive and specific measurement of sIgA in various exocrine secretions.


Scleroderma (progressive systemic sclerosis): progress and course based on a personal series of 118 cases.

A follow-up study has been made of a personal series of 118 cases of patients with scleroderma (progressive systemic sclerosis) followed for periods up to 25 years. These were classified according to the early distribution of skin changes as Type 1 (skin changes in fingers only), 48 cases; Type 2 (skin changes beyond the fingers but mainly in extremities), 47 cases; and Type 3 (diffuse), 19 cases, and atypical, four cases. Eighty per cent of the total series have survived five years, and 68% have survived 10 years. Survival is related to the type of scleroderma, being much better for Types 1 and 2 (acrosclerotic) than for Type 3 (diffuse). Actual compared with expected survival rate was better for younger than for older patients and for females than for males. Although the disease type gives an indication of survival in general, the caution must be used in the individual cases, as the course of the disease is very variable even in patients of the same type, and is subject to remission and relapse.


[Human anthrax in Tougan district (Upper-Volta) (author's transl)].

23 cases of human anthrax observed in a limited district of Upper-Volta are reported with comments on clinical and epidemiological features and treatment.


[A new pathophysiological explanation of ciguaterra (author's transl)].

After pointing out clinical, biochemical and biological similarities between paralytic shellfish poisoning and ciguaterra fish poisoning, the authors describe their recent discoveries in regard to ciguaterra origin. The causal agent searched without results since many years, might be a dinoflagellate, of the genus Diplopsalis.


Role of cell-associated pathogen metabolism in infection of tracheal explants by Mycoplasma pneumoniae.

The effect of environmental conditions on the relative pathogenicity of Mycoplasma pneumoniae for hamster tracheal explants was investigated. Organisms from the early stages of the growth cycle (e.g., day 1 to 2) were more effective in the induction of ciliostasis than were older cultures. Both the degree of ciliostasis and the speed of onset were affected. The type of explant culture medium also affected pathogenic potential. M. pneumoniae infection produced significantly greater ciliostasis and cytonecrosis in a "permissive" medium, i.e., one capable of supporting mycoplasma metabolism and replication, than in a "nonpermissive" medium. However, no adenine protection effect could be detected under permissive conditions, though it was quite striking when a nonpermissive medium was used as the post-infection explant medium. This suggests that the cell damage noted under permissive conditions may result from processes distinct from those operative in the actual host-parasite cellular interaction.


Growth of some attenuated influenza viruses in hamster tracheal organ cultures.

Hamster tracheal organ cultures were infected with different influenza viruses and the metabolic activity measured using a tetrazolium reduction assay. In addition, relative ciliary activity was observed, virus multiplication measured, and histological studies were made. The hamster organ cultures proved relatively simple to set up and were sensitive to infection with influenza viruses; and the tetrazolium reduction was a reliable objective measure of relative tissue damage correlating well with histological and virological findings as well as with visual assessment of ciliary activities.


Somatic crossing-over in Glycine max (L.) merrill: activation of dimethyl nitrosoamine by plant seed and comparison with methyl nitrosourea in inducing somatic mosaicism.

The soybean system used for detecting environmental mutagens is analyzed for various types of spots on the leaves of heterozygous y11y11 plants and homozygous y11y11's induced by a nitrosoamine (dimethyl nitrosoamine, DMN) and a nitrosoamide (methyl nitrosourea, MNU). It is shown that the nitrosoamine can be "activated" by the seed (is converted to a true mutagen) without the addition of NADPH or S-9 fraction of the liver homogenate as is necessary in animal tissue culture or bacterial studies. Whereas somatic mosaicism in soybean can be induced with a dose as low as 1.25 ppm of DMN, the upper limit in spot production is reached at around 60 ppm concentration, applied for 0--24 h. Such saturation effect may be due to a limited amount of DMN being converted to true mutagen. MNU, on the other hand, does not show such limitations, perhaps because of its property of being a direct mutagen not necessitating an intermediate step required for converting the promutagen DMN. The frequency of twin spots on Y11y11 leaves increases only slightly by either DMN or MNU, suggesting only a small increase in somatic crossing-over induced by the two chemicals. The yellow spots increase the most, perhaps due to segmental losses carrying Y11 or non-complementary segregation of exchanges involving non-homologous chromosomes. Neither chemical is found capable of mutating y11 to Y11 as seen by the general lack of light green sectors on y11Y11 plants. Usefulness of the soybean system in studying mutagenesis is briefly discussed.


Immune thrombocytopenia. Use of a Coombs antiglobulin test to detect IgG and C3 on platelets.

We applied a radiolabeled Coombs antiglobulin test to the diagnosis and management of immune thrombocytopenia in adults and children. This assay substantiated that the majority of patients with idiopathic thrombocytopenic purpura have increased levels of IgG on their platelets. Platelet-associated C3 was elevated in a subset of these patients, some of whom had normal levels oet-associated IgG, thus suggesting a role for C3 in the pathogenesis of the thrombocytopenia. The assay enabled us to anticipate a change in clinical status, establish the presence of immunologic remission after splenectomy and propose several mechanisms by which corticosteroids act. Platelets from a patient with the post-transfusion-purpura syndrome also carried increased IgG, indicating a role for IgG antibody or IgG-containing immune complexes in the destruction of host platelets in this disease. The radiolabeled Coombs test provides a general means to help diagnose, manage and study immune platelet disorders.


The effect of chronic ethanol intake on the growth and spread of some murine tumors.

The effect of chronic ethanol intake on the growth and spread of some murine tumors has been investigated. The treatment had no effect on the B 16 melanoma but tended to decrease the number of Ehrlich ascites cells. In the case of the Lewis lung carcinoma, administration of ethanol for two weeks tended to lower the number of metastases to the lung without significantly affecting the primary tumor size, whereas more prolonged ethanol intake decreased the weight of the primary tumor in addition to decreasing its dissemination.


[Orientation of the cleavage spindles in pulmonate mollusks. I. The role of the form of the blastomeres in 2d cleavage spindle orientation].

In the normal two-celled embryos of various pulmonate molluscs, the orientation of spindles characteristic of metaanaphase is being frequently established gradually, in the process of transition from pro- to metaphase accompained by the growth of spindle and asters. The typical growth of contact zone between the blastomeres of the common pond snail embryos was inhibited to a different extent under their cultivation after the 1 cleavage division in the calcium-free media or after trypsinization. At the same time the orientation of meta-anaphase spindles was markedly affected (as judged by an angle alpha between the spindle axis and the plane of contact zone in the equatorial projection). When analyzing the model distributions of the angles between the two spindle axes (in the same projection), it was shown that the empirical distributions of these angles corresponded to the principle of stochastic combination of two alpha. A conclusion is drawn that the orientation of one spindle does not depend on that of another but the position of each of them depends on the size of the contact zone and, hence, on the general form of the adjacent blastomere region. Some other processes determining the spindle orientation are discussed.


[Orientation of the cleavage spindles in pulmonate mollusks. II. The role of the architecture of the intercellular contacts in III and IV cleavage spindle orientation].

In the normal development of pulmonate molluscs, the variety of orientations of the III and IV cleavage spindles markedly reduces in the process of transition from pro- to meta- and anaphase. Even prior to the completion of spindle growth in these cleavage divisions the external faces of blastomeres become asymmetrical due to intercellular interactions but the whole system of external intercellular contacts (edges) is characterized by a certain symmetry. This symmetry coincides partially or fully with that of the system of definitive spindles. In the artificially obtained four-celled common pond snail embryos with the chain-like position of blastomeres, the III cleavage spindles were oriented at right angles to the polar axis, rather than in parallel with it (as in the normal development). The eight-celled embryos with symmetrical external faces of macromeres were also obtained. The variety of orientation of the IV cleavage definitive spindles in such embryos was markedly widened and in the macromeres with inverse asymmetry the inversion of the sign of declination of the spindles was observed. The spindle orientation depends, thus, on the form of adjacent region of the external face. This form as a whole is determined by the mutual position of blastomeres, curvature of surface and relative length of the face edges.


[Action of embryonic thymus and liver cells on the development of the ascitic form of Ehrlich's carcinoma].

The influence of the cells of embryonic thymus and liver on the development of Ehrlich carcinoma was studied. The intraperitoneal injection of the embryonic cells in the adult mice infested by the Ehrlich carcinoma resulted in a marked lengthening of the life time of animals and an increase of the survival percentage. The embryonic cells of thymus and liver inhibited sharply the growth of carcinoma cells in the diffusion chambers as well. In contrast to this, the thymus and bone marrow cells of adult animals, taken in the same concentrations as the embryonic cells, exhibited only a slight inhibiting effect on the growth of tumour cells. On the basis of these data a suggestion is put forward to the effect that the embryonic immunocompetent cells determine the stronger inhibition of tumour growth in the embryos as compared with the adult animals.


Cardiovascular emergencies in pediatrics.

This article dicusses emergencies of cardiovascular origin in infants and children. Particular attention is given to congestive heart failure, hypoxic states, arrhythmias, cardiac tamponade, perinatal cardaic emergencies and conditions simulating cardiac crises. An outline of etiologic and therapeutic approach is included, and guidelines are provided for the practitioner who is called to care for such emergency situations.


Social pediatric emergencies.

A social pediatric emergency invokes the concept of a crisis situation, which often reflects an acute temporary state, the culmination of problems of long duration. The needs demonstrated in child abuse, neglect and deprivation, sexual abuse, the handling of the crisis of birth, pregnancy and abnormality, death and dying, adoption and learning disabilities are related to the family psychodynamic relationships and the doctor team approach. The social pediatrician can play his role as the physician, advocate, activist and educationalist in the many complex situations revealed within family, community and society.


Energetics of anaerobic glycolysis in dog gastrocnemius.

Thermally isolated gastrocnemii were stimulated to exhaustion, by rhythmic isotonic (70 N) tetanic contractions, during complete occlusion of blood flow. Enthalpy change (h = work + heat) and work output (w) (kJ/kg) were obtained from records of deep muscle temperature and shortening. The lactate produced (LA, mol/kg) was measured in the outflow after reestablishement of blood flow. The following relationships were obtained: h = 76LA + 1.2, and w = 19.8LA + 0.30. As the energy liberated at exhaustion by alactic energy sources (approximately P and O2 stores) is constant, deltah/deltaLA = 76 (+/- 10.5; S.E.) kJ/mol is the enthalpy change for lactate formation (delta HLA). The neutralization heat was estimated on muscle homogenates at 12kJ/mol, leaving approximately 64 kJ/MOL for deltaH of LA formation proper. The mechanical efficiencies of work related to LA formation (ELA) and of that not related to LA formation (EnonLA) were practically identical (0.25). From these values and from deltaHLA, the enthalpy change of approximately P splitting was estimated in the range of 52--62kJ/mol, depending on the value of the ratio delta approximately P/deltaLA assumed in the calculation.


Quantitative description of acetylcholine release and fluctuations in nerve terminals of torpedo electric organ submitted to stimulation.

A model, considering cytoplasmic ACh as the difference between cumulated outputs per impulse, and synthesized ACh leads to simple equations correlating biochemical and electrical measurables. Intrinsic properties of the release mechanism are deduced.


The influence of high ambient temperature on thermoregulatory response to intrahypothalamic injections of noradrenaline and serotonin in the pigeon.

1. Pigeons with chronically implanted injection cannula in the hypothalamus were injected with noradrenaline (NA, 10 micrograms/microliter) and 5-hydroxytryptamine (5-HT, 10 micrograms/microliter) at the ambient temperature (Ta) of 38 degrees or 42 degrees C. 2. Initial tests at cold (6 degrees C) indicated that birds responded hypothermically to NA and in most instances also to 5-HT. 3. Intrahypothalamic injection of NA (10 micrograms/microliter) had no appreciable effect on oxygen consumption (VO2), body temperature (Tb), foot temperature (Tf), or heart and respiratory rates at Ta 38 degrees C. The increase of VO2, Tb and Tf noted after similar injection at 42 degrees C was in all probability due to observed excitement and bursts of struggling rather than effects on thermoregulatory mechanisms. 4. 5-HT (10 micrograms/microliter) injected at Ta 38 degrees C depressed respiratory frequency from panting (600 breaths.min-1) to normal rate (ca. 50.min-1) within 2--4 min. The absence of panting lasted about 10 min, but only a slight increase of VO2, Tb and Tf followed. At Ta 42 degrees C, no notable changes of VO2, Tb and Tf were recorded after 5-HT injection. 5. It is concluded that 5-HT has an inhibitory action on neuronal pathway controlling panting activity in the pigeon, but NA seems to be ineffective.


Effect of divalent cations on acetylcholine release from cardiac parasympathetic nerve endings.

The influence of several divalent cations on acetylcholine (ACh) release from vagus nerve terminals was studied in quiescent guinea-pig auricles. ACh release was induced by stimulus trains subthreshold for excitation of atrial cells. The ACh released evoked a hyperpolarization of the atrial cell membrane. Changes in ACh sensitivity of the atrial cell membrane were tested by application of ACh-containing solutions. ACh release increased with the external Ca concentration. External Sr maintained the ACh release in 'Ca-free' media. However, Sr ions were less effective than Ca ions. External Mg acted as a weak inhibitor of the stimulus evoked ACh release, whereas Co and Mn ions exerted a strong inhibition.


Micropuncture study on the effects of lithium on proximal and distal tubule function in the rat kidney.

Micropuncture studies were performed in rats infused with LiCl to induce stable plasma lithium concentrations of 2--3 mEq/l, or with an equivalent amount of NaCl. In free flow experiments LiCl reduced proximal tubule fractional reabsorption of sodium and potassium. Reduced reabsorption of bicarbonate, as reflected by a decrease in TF/PCl, was also observed. Proximal fractional reabsorption of chloride, however, was not affected. The TF/PIn at the end proximal tubule was 2.6 +/- 0.2 (mean +/- SEM) in controls and 2.1 +/- 0.1 in the experimental animals (P less than 0.025). In the distal portions of the nephron lithium treatment caused a fall in fractional reabsorption of water and sodium, while potassium secretion was stimulated in the distal tubule. Previous studies have indicated that lithium influences antidiuretic hormone stimulated water transport in the collecting duct. These experiments demonstrate that lithium also affects the transport of water and electrolytes in multiple nephron segments, including the proximal and distal convolution.


Effects of vasopressin and its deamino-D-arginine analogue on renin release in the isolated perfused rat kidney.

The direct action of arginine-vasopressin (AVP) and its deamino-D-arginine analogue (DDAVP) on renin release (RR) has been studied in isolated rat kidneys perfused with an electrolyte solution at constant pressure in a single-pass system. AVP and DDAVP infused at various concentrations (80 to 2100 pg/ml and 80 to 8700 pg/ml, respectively) reduced volume and increased osmolality of urine in a dose-dependent way. High doses of AVP reduced renal perfusate flow and glomerular filtration rate while DDAVP had no effect on renal haemodynamics. When vasoconstrictor doses of AVP or high concentrations of DDAVP were infused, "basal" RR remained unchanged. However, when RR had been stimulated by infusion of isoproterenol, vasoconstrictor doses of AVP as well as high doses of DDAVP which did not increase renal vascular resistance diminished RR by about 30% (P less than 0.01, and P less than 0.05, respectively). These results suggest that the inhibition of RR by vasopressin is not related to its vasoconstrictor action.


The differential effect of cooling on the responses of splenic capsular and vascular smooth muscle to nerve stimulation and noradrenaline.

The responses of the capsular and vascular smooth muscle of the dog's spleen to splenic nerve stimulation and to infused noradrenaline have been studied in the isolated, blood-perfused preparation at 37 degrees C, at 27 degrees C and again after rewarming to 37 degrees C. It was found that cooling per se had no effect on perfusion pressure but reduced splenic arterial blood flow, and caused no appreciable alteration in spleen volume. The increase in splenic flow resistance in response to nerve stimulation and noradrenaline was significantly greater at 27 degrees C than at 37 degrees C, but the concomitant reduction in spleen volume was significantly reduced. The enhanced effect of splenic nerve stimulation and noradrenaline on splenic flow resistance is discussed in terms of the relative contributions of an increased smooth muscle response and increased blood viscosity. The different effects of cooling on the responses of splenic vascular and capsular smooth muscle to nerve stimulation and noradrenaline are discussed in the context of the effect of cooling on other vascular and non-vascular smooth muscle.


Maximal reabsorptive capacity for inorganic phosphate (TmPi) in the absence of parathyroid hormone in the rat: decrease of the TmPi during prolonged administration of phosphate and the role of calcium.

Clearance titration studies were performed to examine the maximal reabsorptive capacity for inorganic phosphate (TmPi) in acutely parathyroidectomized (PTX) rats. Under control conditions, the reabsorption of Pi was 6.01 +/- 0.92mumol/min (mean +/- S.D.), and during the administration of phosphate the reabsorption of Pi increased initially to 11.62 +/- 2.20 and decreased finally to 7.97 +/- 2.02mumol/min. The Reabs/GFR values were 2.12 +/- 0.23 (control), 3.57 +/- 0.21 and 2.49 +/- 0.42mumol/ml (experimental), respectively. Thus, the TmPi decreased significantly during phosphate loading. Because this drop of the TmPi was accompanied by a significant decrease of the calcium concentration in plasma in these PTX animals, another series of experiments was performed in which the calcium losses in plasma were replaced. Under these conditions the decrease of the TMPi was significantly smaller than in hypocalcemia. It is concluded that calcium may regulate the renal Pi transport in the absence of parathyroid hormones, at least in hypo- and normocalcemic states.


Magnesium handling in the dog kidney: a micropuncture study.

Micropuncture and clearance experiments were done on normal dogs to investigate magnesium handling by proximal and distal nephron segments. Tubular fluid electrolytes were analyzed with the electron microprobe. Tubular fluid to ultrafilterable magnesium ratio (TF/UF magnesium) was observed to rise above unity but less than the TF/P insulin ratio generated along the proximal tubule. This is in contrast to the other major cations, the ratios of which remain close to unity as water is abstracted. Tubular fluid obtained from the distal tubule contained less magnesium than the glomerular filtrate (mean TF/UF magnesium of 0.6) indicating the loop of Henle is the major nephron segment reclaiming a significant portion of the filtered load. The faction of filtered load remaining at the distal sampling site was similar to the fraction appearing in the urine (8% vs 7%) indicating very little reabsorption beyond the distal tubule in these normal states.


[Recent Lassa, Marbourg and Ebola viruses in African tropical viruses. I. Semiology--physiopathology--diagnosis--treatment (author's transl)].

Three new viruses have been identified in Africa during the present decade. They may cause sporadic cases or limited outbreaks, and they are probably endemic in areas which are still ill-defined. Severe forms of infection lead to the haemorrhagic syndrome or to hypovolemic shock, the physiopathology of which is being studied. The case-fatality ratio of severe cases is between 30 and 85 per cent. Nosocomial outbreaks have been observed, but they can be avoided if appropriate barrier nursing measures are carried out for the treatment of patients or adequate protection measures for sampling and examination of laboratory specimens. As such cases may be transferred outside the endemic zone, this implies that countries receiving travellers from Africa should have hospitals with specialized units for strict isolation and treatment of these patients.


Localisation of foldback DNA sequences in nuclei chromosomes of Scilla, Secale, and of mouse.

Foldback DNA, prepared from mouse and Scilla sibirica main band DNA, and from rye (Secale cereale) total DNA, was characterised by denaturation, renaturation, and electron microscopy. 3H-cRNA of this DNA was hybridised in situ to nuclei and chromosomes of the respective species. There is no universal labelling pattern among the three species. In mouse, highly repetitive foldback DNA is present in the whole chromatin including the satellite DNA-containing regions. In Scilla sibirica, on the contrary, the highly repetitive foldback sequences are excluded form the satellite DNA loci and are arranged in clusters in the remaining chromatin. In rye, there is a clear preferential labelling of the chromocenters in the interphase nuclei as well as metaphase chromosomes, indicating that highly repetitive foldback DNA is preferentially located among other highly repetitive sequences.


DNA polymerase alpha and beta in the California urchin.

DNA polymerase alpha and beta were identified in the urchin, Strongylocentrotus purpuratus. The DNA polymerase beta sedimented at 3.4 S, constituted 5% of total DNA polymerase activity, and was resistant to N-ethylmaleimide and high ionic strength. The polymerase alpha sedimented at 6--8 S, was inhibited by N-ethylmalemide or 0.1 M (NH4)2SO4, and was dependent upon glycerol for preservation of activity. Both the polymerases alpha and beta were nuclear associated in embryos. The DNA polymerase alpha was markedly heterogeneous on DEAE-Sephadex ion exchange and showed three modal polymerase species. These polymerase alpha species were indistinguishable by template activity assays but the DNA polymerase associated ribonucleotidyl transferase (Biochemistry 75 : 3106-3113, 1976) was found predominantly with only one of the DNA polymerase alpha species.


Preparation of a complementary DNA for leghaemoglobin and direct demonstration that leghaemoglobin is encoded by the soybean genome.

In soybean root nodules, leghaemoglobin (Lb) accounts for 25--30% of the total soluble protein but is not detected in other tissues. In order to determine whether the Lb genes are plant or bacterial in origin a cDNA probe for Lb was prepared from 9S poly (A) containing mRNA of root nodules. Although this 9S mRNA directed synthesis of predominantly three forms of Lb in vitro, the kinetics of hybridisation of cDNA and the 9S mRNA showed a transition at about 30% hybridisation which suggested that the 9S-cDNA was not pure Lb-cDNA. The abundant, Lb-cDNA was prepared by two cycles of hybridising 9S mRNA and cDNA to a Rot of 3 X 10(-3) and isolation of the hybridised cDNA on hydroxyapatite. The Lb-cDNA was homogeneous in hybridisation analysis with 9S mRNA and electrophoresis in 98% formamide gels. This cDNA hybridised with soybean DNA and not with Rhizobium DNA, thus directly demonstrating that Lb genes are of plant origin. Titration of Lb-cDNA with soybean DNA showed that Lb genes are reiterated about forty-fold per haploid genome.


Partial purification of transcriptionally active nucleosomes from trout testis cells.

Mononucleosomes (MN1) enriched in structural non-histone proteins and transcribed DNA sequences were obtained by limited digestion of trout testis nuclei with micrococcal nuclease followed by selective solubilization in 0.1 M NaCl. These monosomes consist of the four inner histones plus stoichiometric amounts of the non-histone protein H6, of the HMG group, complexed with 140 base pairs of DNA. Hybridization experiments indicate that MN1 DNA is enriched in sequences complementary to cytoplasmic polyadenylated RNA.


[Studies of lipase-induced fat necrosis in rats (author's transl)].

Disseminated fat necrosis can be produced by intraperitoneal injection of porcine pancreatic lipase. They get detecable by intravital staining with Phosphine 3R about 15 min after injection. The earliest fine structural findings are spotty destruction of the pinocytic invaginations and vesicles, combined with alterations of the cell membrane. Later there is a complete destruction and disintegration of the cytoplasm and its organels as well as the nucleus, whereas the cell membrane partly remains visible. At the same time the central lipid droplet shows cloudy disintegration and clumping as well as cristalline and granular structures. In the beginning the necrosis is limited to single cells. Later the adjacent fat cells also undergo necrosis, even when the applicated lipase has been removed after 30 min. In light microscopic studies fat necrosis are detectable 30 min after the application of lipase. During the first 48 hours they get demarcated by leucocytes. In the following days resorption and organisation take place. Lipolytic drugs facilitate the development of fat necrosis, whereas antilipolytic drugs inhibit it. In starvation the number of fat necrosis rises, after feeding it decreases. In the diurnal rhythm there is a maximum after midnight and a minimum in the early afternoon. The results support the hypothesis that pancreatic lipase only attacks fat cells, which are lipolytically active.


Growth deficiency dysmorphic syndromes.

The precept is set forth that many patterns of malformation are the consequences of congenital hypoplasia in the skeletal system, as in osteochondrodysplasias, or in multiple tissues as in a host of dysmorphic syndromes. The disorders include those caused by most known teratogens in man, by chromosomal abnormalities, mutant gene disorders and a number of syndromes of unknown etiology. Not uncommonly there is a good correlation between the degree of linear growth deficiency and brain growth deficiency within a particular syndrome. Postnatally there is usually a lack of catch-up growth; the children tend to grow at a normal rate for their diminished size by birth. Because there is often a problem in skeletal morphogenesis, roentgenographic interpretation for 'bone age' may be misleading in terms of maturational age. There is no known therapy for congenital hypoplasia disorders, and the ultimate prognosis for height may be judged from the height of adults with the particular disorder in question and related to genetic background of the parents. For example, the adult growth of girls with XO Turner syndrome tends to be about 3.7 SD's below that expected for mean parental height.


The fetal brain and intrauterine growth.

The fetal brain stimulates intrauterine growth in man, rat and other species. Human congenital brain anomalies and experiments in rats indicate that the hypothalamus is the brain area responsible for this effect. The only factor found to stimulate intra-uterine growth was alpha-MSH. Various observations and experiments in rat show that stimulation of fetal somatic growth might indeed be a physiological function of alpha-MSH, while observations in the human fetus indicate that alpha-MSH might play a similar role in intra-uterine growth in man as well. Preliminary data in rat point to a similar function of this hormone in brain development.


[Tinea nigra. Apropos of a case diagnosed as melanoma of superficial dissemination].

The authors report a case of tinea nigra of left sole, diagnosticate at the beginning as melanoma of superficial dissemination. The mycological studies confirmed the dermatological diagnostic of tinea nigra by Cladosporium werneckii.


[Effect of different nitrogen sources on the biosynthesis of lipase by Oospora lactis].

The effect of inorganic and organic nitrogen compounds on the synthesis of biomass and extracellular lipase by Oospora lactis was studied. Among the inorganic nitrogen sources ammonium sulphate and ammonium secondary phosphate and among the organic nitrogen sources yeast autolysate proved to be most beneficial for the lipase synthesis. Lipase activity and biomass accumulation in the medium containing yeast autolysate were greater than in the media containing the above ammonium salts. Lipase synthesis reached maximum in the nutrient medium containing yeast autolysate (0.7%) and ammonium sulphate (0.3%).


An evaluation of the intrapair relationship for personal attributes in the NHLBI twin study.

We have considered a group of questions which attempt to define the environments of twins in our study both within and between twin pairs. Only 8 of 31 questions on which twins compared themselves to their co-twins were reliable and showed discordance for twin pairs. Comparing discordant twins for cardiovascular risk factors, few significant results were obtained. Cigarette smoking seemed to be most closely related to the environmental differences; but negatively related to achievement in grade school and high school and to which twin was stronger in childhood. Weight at examination appeared to be related to strength in childhood. In addition, there may be a negative relationship between income and blood pressure. One additional question showed MZ twins get together significantly more often than DZ twins. In terms of differnces in environmental covariance, the question "how frequently do you and your twin get together now?" may be a simple but valuable measure of similarity of twins and should be considered for inclusion in twin studies.


Synthesis of some 2-styrylquinazoline derivatives structurally related to certain chemotherapeutic agents.

The synthesis of certain 2-styryl-4-arylaminoquinazolines having the general formula 1 is described. 4-Chloro-2-styrylquinazolines were prepared by chlorination of 2-styryl-4-quinazolones in benzene solution using phosphorus oxychloride and an excess of dimethylaniline. Reacting the chlorostyrylquinazolines with certain arylamines afforded 1.


Investigation of iridoid substances in Arctostaphylos uva-ursi.

Active principles of bearberry have been studied for several decades. The drug Folium uvae-ursi is useful to the pharmaceutical industry (in production of medicinal teas) for its disinfective effect on the urinary tract which is derived from the content of phenolic glucosides. This work is a part of the total evaluation of the drug from the point of view of contents of active principles, biogenesis, isolation, qualitative and quantitative differences during the vegetative period, and the changes during technological treatment and pharmacological activity of its active principles.


Effect of electrostatic fields on the chromosomes of Ehrlich ascites tumor cells exposed in vivo.

An effect of electrostatic fields on the chromosomes of Ehrlich ascites tumor cells exposed in vivo has been demonstrated. Cells exposed to horizontal electrostatic fields for two weeks had almost a threefold increase in the percentage of abnormal chromosomes when compared to control cells or cells exposed to vertical electrostatic fields for the same period. Extended exposure times of 4--15 weeks resulted in the disappearance of the aberrant chromosomes. It is suggested that the effected cells were incapable of cellular replication resulting eventually in their disappearance via cell death.


A fluorescent probe study of salmine AI.

A fluorescent probe, 1-p-toluidinylnapthalene-8-sulfonate (1,8-TNS), was used to study the nonpolar sites on salmine AI. Fluorescence enhancement resulting from binding between the probe and the protein occurs at a wavelength of maximum emission of 497-500 nm, indicating the existence of moderately nonpolar binding sites on salmine AI. Fluorescence enhancement decreases as the ionic strength of the solvent is increased from 0.002 M to 0.050 M. Fluorescence increases with increasing acidity although this effect is not correlated to the pKa of 1,8-TNS. Positive cooperative binding takes place between 1,8-TNS and salmine AI. Equilibrium dialysis indicates that binding occurs only under conditions resulting in significant fluorescent enhancement. The binding was also studied using thin film dialysis, which is much faster than equilibrium dialysis and avoids the observed changes in probe-protein interaction that occur over long time periods with the latter system.


Ultrasonographic evaluation of vascular access complications.

In 24 patients, sonograms were obtained of 25 vascular accesses, 16 of which were bovine grafts, 7 arteriovenous fistulae, 1 a polytetrafluoroethylene graft, and 1 an external Silastic shunt. On the sonograms, the lumen, course, and insertions of the vascular grafts and fistulae were outlined, anterior and posterior aneurysms demonstrated, hematomas differentiated from aneurysms, and intrinsic distinguished from extrinsic lesions. The authors conclude ultrasonography is a valuable adjunct to clinical evaluation in assessing certain complications of vascular access.


Changes in the effects of cocaine during chronic treatment.

The principle object of the study was to identify the changes in the rat's response to cocaine which occurred during chronic administration of the drug. Doses of 20 and 35 mg/Kg cocaine were administered at intervals of 12 and 24 hours, for 300 days or until their effects proved fatal. Four main effects were observed: sensitization to cocaine's convulsant and behavioral activating properties, tolerance to its weight loss effects and decreased response duration. Convulsant sensitization occurred primarily among rats receiving the higher dose of cocaine. All other effects occurred with equal frequency in rats at both doses. None of the changes were dependent on the interval between injections.


Effect of propranolol on glycerol induced acute renal failure in rats.

The effect produced by propranolol, administered for a prolonged period of time and in large doses, on renal function in rats has been studied, as well as the modifications induced by this treatment in an experimental model of acute renal failure, and the effects of a single dose of propranolol given 1 hour before provoking failure. Propranolol, administered chronically, causes sodium and water retention and increases creatinine clearance. Acute renal failure induced by glycerol in rats treated for 7 days with propranolol is less severe than the one produced in untreated animals. In this ARF model, a single dose of propranolol does not seem to have a protective effect.


Urate metabolism and gastric ulcerations in rats as influenced by stress and allopurinol.

Stress and allopurinol (ALL) were investigated in rats with regard to their influence on serum uric acid and glucose concentration, gastric secretion, microcirculation (MBF) and stress ulcer index. There is a non-competitive type of interaction between severe stress and ALL-mediated xanthine oxydase inhibition (= per cent fall in serum uric acid) as compared with control conditions (= mild stress). Vmax is different (84.6 +/- 1.9 and 92.3+/-2.26; P less than 0.05), but not Km (0.39 +/- 0.09 and 0.68 +/- 0.08 mg/kg/h ALL). Serum uric acid is higher in rats with draining gastric fistula than those with closed fistula suggesting that already mild stress is associated with an increase in uricemia in this species. ALL does not significantly alter gastric acid and uric acid secretion but improves markedly gastric ulcer index during mild and severe stress. Since MBF is significantly elevated by ALL during the latter circumstances, a dissociation between MBF and acid secretion is one feature of ALL actions and might become a primary aim in treatment of this disorder.


Isolation of mycoplasmas from the canine genital tract: a survey of 108 healthy dogs.

Cultivation of preputial or vaginal swab samples from 108 apparently healthy dogs gave presumptive mycoplasma growth in 72 cases. Specific identification of 58 mycoplasma isolates based on biochemical studies, indirect immunofluorescence and growth inhibition tests showed association with eight different species or groups, only four of which had previously yielded genital isolates. Fourteen strains were related to M bovigenitalium.


Instability of benzimidazole resistance in nematode eggs.

The resistance of the eggs of benzimidazole resistant Haemonchus contortus and Trichostrongylus colubriformis is not stable and will decrease in the absence of drug challenge. Dosing animals harbouring resistant H contortus with either thiabendazole or mebendazole selects for nematodes producing thiabendazole resistant eggs. The eggs of a thiabendazole selected strain of H contortus were more resistant to thiabendazole, parbendazole and oxibendazole but less resistant to cambendazole and mebendazole.