[Preschool age mortality (1-4-year olds) in Cuba, 1961-1975]. An abstract is made of the preschool age mortality in Cuba through the period of 1961 to 1975. The authors outlight how it has dropped from 2.0 to 1.1 per 1000 population to reach 45% reduction. They establish comparison with a group of American countries, setting of contrast with the low mortality in Cuba. The mortality in children of 1 to 4 years old is considered as indirect index of malnutrition. The percentage of diminution for the different departments in the period stated is as follows: Pinar del Río, 57.6%; Havana, 43.7%; Matanzas, 58.5%; Las Villas, 63.1%; Camagüey, 50.0%; Oriente, 39.0%. The 10 most important causes of death are analyzed and the percentages in drop of mortality are reported as follows: (A 43), 80.0%; (A65) and (A105-106), 66.7%; (A5), 61.5%; (A93), 50.0%; (A89, 90, 91 and 92), 46.5%; (A21), 42.8%; (AE 138-146), 34.6%; (A126-130), 31.2%; (A25), 25.0%. Finally, stress is placed on the elements responsible for the drop in preschool age children's mortality.


Haemodynamic effects of hydrallazine and of hydrallazine plus glyceryl trinitrate paste in heart failure. In a study designed to investigate potential non-parenteral treatment for chronic heart failure, hydrallazine, 225 to 300 mg per day, was given orally to 9 patients. There was no significant change in heart rate or mean arterial pressure as cardiac output increased. Left ventricular stroke work increased significantly and pulmonary artery wedge pressure fell. Systemic and pulmonary vascular resistances fell. With the addition of 2 per cent glyceryl trinitrate paste, there was a further decline in mean pulmonary arterial and wedge pressures, without a significant change in heart rate, arterial pressures, cardiac output, or systemic or pulmonary vascular resistance. There were no untoward effects from either form of treatment. All patients reported relief of shortness of breath and other symptoms related to ventricular dysfunction. This study supports the suggestion that oral hydrallazine is effective in increasing cardiac output and decreasing pulmonary congestion. Furthermore, the addition of topical glyceryl trinitrate provides a greater reduction of pulmonary pressures, probably through its predominant venodilator action. In some selected patients with heart failure, oral hydrallazine and topical glyceryl trinitrate in combination produce beneficial clinical and haemodynamic effects, probably through afterload and preload reduction, respectively.


Implications of changes in amplitude and contour of the mercury strain gauge plethysmograph pulse tracing. Alterations in the mercury strain gauge plethysmograph pulse tracing have been previously thought to denote changes in venous tone. This study was designed to define the implications of changes in amplitude and contour of pulse tracing. Seven normal subjects were studied. After control measurements, amyl nitrite was administered and the pulse tracing was recorded. When the haemodynamic state had returned to control levels 0.4 mg sublingual glyceryl trinitrate (GTN) was given. The amplitude of the plethysmograph pulse tracing (Y mm) and the vertical distance from the dicrotic notch to the peak of the pulse tracing (X mm) were measured. With amyl nitrite, the amplitude of the pulse tracing (Y) decreased from 10.4 +/- 1 to 4.5 +/- 0.6 mm (mean +/- SEM), while the X/Y ratio increased from 0.7 +/- 0.02 to 0.98 +/- 0.02 (P less than 0.005). After GTN Y increased from 9.3 +/- 0.8 to 31 +/- 4.8 mm (P less than 0.005) and X/Y ratio increased from 0.7 +/- 0.06 to 0.99 +/- 0.01 (P less than 0.005). Since it is known that amyl nitrite produces venoconstriction and arteriolar dilatation, and GTN causes venous and arteriolar dilatation, we conclude that an increase in Y reflects venous dilatation, and an increase in X/Y ratio, that is a fall in the dicrotic notch, denotes arteriolar dilatation. Thus, the plethysmograph pulse tracing can be used to assess the effects of drugs on the capacitance and resistance beds.


The effect of intravenous heparin infusions on the thyroid stimulating hormone response to thyrotrophin releasing hormone. 1 The TSH response to TRH is suppressed 24 h after the commencement of i.v. heparin infusion in euthyroid subjects, as compared to repeat testing, both after 7 days of continuous i.v. heparin and after 7 days without heparin but while on warfarin. 2 This suggests that the rise in serum free thyroid hormones, which heparin produces, has a 'metabolic effect'. Possible clinical effects of this action must be considered. 3 Heparin has no effect on the radio-immunoassay of TSH.


In vitro evaluation of some derivatives of the carcinogen Butter Yellow: implications for environmental screening. The rat-liver carcinogen 4-dimethylaminoazobenzene (Butter Yellow, DAB) and 12 of its structural analogues have been evaluated in a cell transformation assay. Eight of these analogues have already been tested for carcinogenicity in rats, whilst the remaining 4 are new or hitherto untested. Benzidine and its 3,3'-disulphonic acid derivative have also been evaluated. The in vitro results agree with long-term animal data for 8 compounds but disagree in finding DAB-4'-sulphonic acid, 4-trifluoromethyl-DAB and 4-diethylaminoazo-benzene positive. Possible reasons for these divergencies are discussed. It is concluded that 9-phenylazojulolidine and N-methyl-5-phenylazoindoline have carcinogenic potential and that 3,5-dimethyl-4-aminoazobenzene and 4-aminoazobenzene-4'-sulphonic acid are likely to prove non-carcinogenic. Addition of azobenzene to the in vitro assay medium increases the transforming potency of DAB 25-fold. It is suggested that it acts as a competitive substrate for one of the enzymes that detoxify DAB, and that this effect is related to that produced by norharman. Sulphonic-acid derivatives of established carcinogens are usually inactive. The basis of this effect has been investigated, and it is suggested that it can operate by two separate mechanisms. It has been established that this assay cannot be relied upon to predict the in vivo potency of a carcinogen. Consideration has been given to possible changes which could be made to the liver activation system (the S-9 mix) currently used in in vitro carcinogenicity assays, and a diagram is presented of the metabolic conversions of a compound which might lead to mutation or tumour formation. This enables the term potential carcinogen to be accurately defined, and indicates a possible difference between absolute non-carcinogens and compounds which fail to produce cancer in vivo.


Oligonucleotides in heterogeneous nuclear RNA: similarity of inverted repeats and RNA from repetitious DNA sites. A comparison has been made by oligonucleotide analysis of three fractions of HeLa cell hnRNA: (1) the "snap-back" fraction (ds-hnRNA, 5% of the total); (2) the fraction that self-anneals during prolonged incubation (25% of total); and (3) the fraction that hybridizes most rapidly to an excess of HeLa cell DNA (rep-hnRNA, 10% of the total). T1 fingerprints of each of these hnRNA fractions were similar to one another and featured the largest T1 oligonucleotides of known sequence previously isolated from ds-hnRNA (Robertson, H.D., et al. (1977) J. Mol. Biol. 115, 571--590; Jelinek, W. (1977 J. Mol. Biol. 115, 591--602). When hybridized to DNA either in solution or immobilized on filters, the isolated ds-hnRNA and the rep-hnRNA fractions showed similar hybridization kinetics in the COt range of "intermediate" repetitive DNA sequences; the ds-hnRNA and the rep-hnRNA also self-annealed to equal extents in the absence of any DNA. DNA of all buoyant density classes contained the T1 oligonucleotides diagnostic of the ds-hnRNA and the rep-hnRNA. While hnRNA is rich in inverted repeated sequences, cytoplasmic mRNA contains far fewer such sequences.


Accumulation of thallous ions (Tl+) as a measure of the electrical potential difference across the cytoplasmic membrane of bacteria. The accumulation of thallous ions (204Tl+) by intact bacteria was investigated. I conclude that Tl+ is a permeant cation, and that it therefore accumulates in response to the electrical potential difference (delta psi) across the cytoplasmic membrane (interior negative). A comparison with other methods shows that the distribution ratio of 204Tl+ serves as a reasonably satisfactory method for measuring the membrane potential of Streptococcus faecalis. Glycolyzing cells of this organism develop membrane potentials of up to 180 mV. Preliminary experiments with Escherichia coli, especially those with a mutant defective in the proton-translocating ATPase, indicate that the Tl+ distribution also serves as a measure of the membrane potential in this organism. The particular advantage of Tl+ over other indicators of the membrane potential is that the cells need not be pretreated in any way. By use of the Tl+ distribution, it was calculated that respiring cells of E. coli develop a membrane potential of 160 mV with D-lactate and 180 mV with glucose as a substrate, respectively.


Coat protein binds to the 3'-terminal part of RNA 4 of alfalfa mosaic virus. All four RNAs of alfalfa mosaic virus contain a limited number of sites with a high affinity for coat protein [Van Boxsel, J. A. M. (1976), Ph.D. Thesis, University of Leiden]. In order to localize these sites in the viral RNAs, RNA 4 Tthe subgenomic messenger for coat protein) was subjected to a very mild digestion with ribonucleast T1. The ten major fragments, apparently resulting from five preferential hits, were separated and tested for messenger activity in a wheat germ cell-free system, as well as for the capacity to withdraw coat protein from intact particles. Fragments which stimulated amino acid incorporation were assumed to contain the 5 terminus. Strong evidence was obtained for the location of sites with a high affinity for coat protein near the 3' terminus. The smallest fragment which has the 3'-terminal cytosine comprises only 10% of the length of intact RNA 4 but still possesses these sites. Evidence is presented that the complete coat protein cistron is in the complementing 90% fragment. Possibly, the high-affinity sites are entirely located in the 3'-terminal extracistronic part of RNA 4. They will have the same position in RNA 3 and, possibly, also in the other parts of the genome of alfalfa mosaic virus. The need of this genome for coat protein in order to become infectious may therefore find its explanation in the fact that a conformational change at the 3' ends of the genome parts brought about by the coat protein is required for recognition by the viral replicase.


Polymerization of human immunoglobulin M. The repolymerization of human IgM following mild reductive cleavage was studied as a model for intracellular polymer assembly. Repolymerization was found to require the presence of J chain and a disulfide exchanging system which could be furnished either intrinsically by the use of the monofunctional thiol mercaptoethylamine or extrinsically by the inclusion of a protein-mercaptan mixed disulfide, and/or a disulfide exchanging enzyme. The degree of repolymerization was dependent on the extent of monomer reduction and the product covalently incorporated one J chain per five monomer units. Disulfide exchanging enzyme probably served as a source of mixed disulfides rather than as an enzymatic catalyst of the reaction. These results are discussed in terms of a tentative mechanism for IgM polymerization.


Purification and characterization of canine alpha-1-antiproteinase. The principal canine plasma protease inhibitor, alpha-1-antiproteinase, has been purified 90-fold with a 25% yield to apparent homogeneity. The purification scheme includes anion-exchange chromatography, to separate away the bulk of the serum albumin; affinity chromatography by insolubilized concanavalin A, to remove most of the other serum proteins as well as traces of albumin; and, finally, sizing on Sephacryl-S-200. Unique to this purification scheme is the batch use of insolubilized hemoglobin--Sepharose beads to remove the ubiquitous contaminant haptoglobin. The purified material has an apparent molecular weight of 58 000, 11.2% carbohydrate, and an E280nm1% = 5.82, and can be separated by isoelectric focusing into at least two distinct forms with pI values of 4.40 and 4.52. In addition, canine alpha-1-antiproteinase is immunologically distinct from human alpha-1-antiproteinase.


Use of fluorescence polarization to monitor intracellular membrane changes during temperature acclimation. Correlation with lipid compositional and ultrastructural changes. Fluorescence polarization of 1,6-diphenylhexatriene (DPH) was used to study the effects of temperature acclimation on Tetrahymena membranes. The physical properties of membrane lipids were found to be highly dependent on cellular growth temperature. DPH polarization in lipids from three different membrane fractions correlated well with earlier freeze-fracture and electron spin resonance observations showing that membrane fluidity progressively decreases in the order microsomes greater than pellicles greater than cilia throughout a wide range of growth temperatures. Changes in membrane lipid fluidity following a shift from high to low growth temperatures proceed rapidly in the microsomes, whereas there is a pronounced lag in the changes of peripheral cell membrane lipids. These data support previous observations that adaptive changes in membrane fluidity proceed via lipid modifications in the endoplasmic reticulum, followed by dissemination of lipid components to other cell membranes. The rapid changes in polarization observed in the microsomal lipids following a temperature shift correspond closely with the time-dependent alterations in both lipid fatty acid composition and freeze-fracture patterns of membrane particle distribution, suggesting that, in the endoplasmic reticulum, lipid phase separation is the primary cause of membrane particle rearrangements.


Rotational relaxation of 1,6-diphenylhexatriene in membrane lipids of cells acclimated to high and low growth temperatures. Measurement of the time-resolved fluorescence depolarization of 1,6-diphenylhexatriene (DPH) in artificial bilayers of microsomal membrane lipids from Tetrahymena gives detailed information concerning the molecular motion of this probe and fluid properties of the membrane lipids which are obscured with steady-state methods. The rotational motion of DPH in these lipids from cells acclimated to 15 and 39.5 degrees C growth temperatures was anisotropic, which agrees with recent time-resolved studies of this probe in synthetic phospholipid systems. Evaluation of DPH polarization data obtained from these lipid fractions at their respective growth temperatures showed differences in physical properties which suggest that "viscosity", per se, of the microsomal lipids is not a strictly regulated as it is in prokaryotic systems. Rotational relaxation of DPH in 39.5 degrees C microsomal lipids measured at 15 degrees C is more complex than that of either lipid fraction measured at its actual growth temperature, suggesting that the probe has partitioned into two dissimilar environments within the bilayer. Similar effects are observed in the microsomes of 39.5 degrees C cells by freeze-fracture electron microscopy following rapid cooling to 15 degrees C. Under these conditions, two distinct regions are observed on the fracture faces, suggesting a correlation between lipid phase changes and alterations in membrane structure.


On the state of carotenoids bound to reaction centers of photosynthetic bacteria: a resonance Raman study. The carotenoids bound to reaction centers of wild, Ga and GIC strains of Rhodopseudomonas spheroides, of Rhodospirrillum rubrum, strain S1 and of Rhodopseudomonas viridis, yield very similar, but unusual resonance Raman spectra. Through a comparison with resonance Raman spectra of 15,15'-cis-beta-carotene, these carotenoids are shown to assume cis conformations, while the corresponding chromatophores contain all-trans forms only. These cis conformations likely are identical for all the carotenoids studied. They remain unaffected by variations of temperature from 20 to 300 K as well as by the redox state of P-870. They are unstable, being rapidly isomerised towards the all-trans forms when extracted from the reaction centers. The possible nature of these conformers is discussed on the basis of their electronic and vibrational spectra.


Functional and structural organization of chlorophyll in the developing photosynthetic membranes of Euglena gracilis Z. II. Formation of system II photosynthetic units during greening under optimal light intensity. The relationships between light-harvesting chlorophyll and reaction centers in Photosystem II were analyzed during the chloroplast development of dark-grown, non-dividing Euglena gracilis Z. Comparative measurements included light saturation of photosynthesis, oxygen evolution under flashing-light and fluorescence induction. The results obtained can be summarized as follows: (1) Photosystem II photocenters are formed in parallel with chlorophyll synthesis, but after a long lag phase. (2) As a consequence, the chlorophyll reaction center ratio (Emerson's type photosynthetic unit) decreases during greening. (3) This decrease is accompanied by considerable changes in the energy transfer and trapping properties of Photosystem II. Most of the initially synthesized chlorophylls are inactive in the transfer of excitations to active photochemical centers and are shared among newly formed Photosystem II photocenters; as a consequence, the number of chlorophylls functionally connected to each Photosystem II photocenter decreases and cooperatively between these centers appears. Results are discussed in terms of chlorophyll organization in developing photosynthetic membranes with reference to the lake or puddle models of photosynthetic unit organization.


Isolation and characterization of photosystem I and II membrane particles from the blue-green alga, Synechococcus cedrorum. Fractions enriched in either Photosystem I or Photosystem II activity have been isolated from the blue-green alga, Synechococcus cedrorum after digitonin treatment. Sedimentation of this homogenate on a 10--30% sucrose gradient yielded three green bands: the upper band was enriched in Photosystem II, the lowest band was enriched in Photosystem I, while the middle band contained both activities. Large quantities of both particles were isolated by zonal centrifugation, and the material was then further purified by chromatography on DEAE-cellulose. The resulting Photosystem II particles carried out light-induced electron transport from semicarbizide to ferricyanide of over 2000 mumol/mg Chlorophyll per h (which was sensitive to 3-(3,4-dichlorophenyl)-1, 1-dimethylurea), and was nearly devoid of Photosystem I activity. This particle contains beta-carotene, very little phycocyanin, has a chlorophyll absorption maximum at 675 nm, and a liquid N2 fluorescence maximum at 685 nm. The purest Photosystem II particles have a chlorophyll to cytochrome b-559 ratio of 50 : 1. The Photosystem I particle is highly enriched in P-700, with a chlorophyll to P-700 ratio of 40 : 1. The physical structure of the two Photosystem particles has also been studied by gel electrophoresis and electron microscopy. These results indicate that the size and protein composition of the two particles are distinctly different.


The mechanism of reduction of the ubiquinone pool in photosynthetic bacteria at different redox potentials. (1) A flash number dependency of flash-induced absorbance changes was observed with whole cells of Rhodospirillum rubrum and chromatophores of R. rubrum and Rhodopseudomonas sphaeroides wild type and the G1C mutant. The oscillatory behavior was dependent on the redox potential; it was observed under oxidizing conditions only. Absorbance difference spectra measured after each flash in the 275--500 nm wavelength region showed that a molecule of ubiquinone, R, is reduced to the semiquinone (R-) after odd-numbered flashes and reoxidized after even-numbered flashes. The amount of R reduced was approximately one molecule per reaction center. (2) The flash number dependency of the electrochromic shift of the carotenoid spectrum was studied with chromatophores of Rps. sphaeroides wild type and the G1C mutant. At higher values of the ambient redox potential a relatively slow phase with a rise time of 30 ms was observed after even-numbered flashes, in addition to the fast phase (completed within 0.2 ms) occurring after each flash. Evidence was obtained that the slow phase represents the formation of an additional membrane potential during a dark reaction that occurs after flashes with an even number. This reaction is inhibited by antimycin A, whereas the oscillations of the R/R- absorbance changes remain unaffected. At low potentials (E = 100 mV) no oscillations of the carotenoid shift were observed: a fast phase was followed by a slow phase (antimycin-sensitive) with a half-time of 3 ms after each flash. (3) The results are discussed in terms of a model for the cyclic electron flow as described by Prince and Dutton (Prince, R.C. and Dutton, P.L. (1976) Bacterial Photosynthesis Conference, Brussels, Belgium, September 6--9, Abstr. TB4) employing the so-called Q-cycle.


Metabolic control of lactose entry in Escherichia coli. A general method has been developed for determining the rate of entry of lactose into cells of Escherichia coli that contain beta-galactosidase. Lactose entry is measured by either the glucose or galactose released after lactose hydrolysis. Since lactose is hydrolyzed by beta-galactosidase as soon as it enters the cell, this assay measures the activity of the lactose transport system with respect to the translocation step. Using assays of glucose release, lactose entry was studied in strain GN2, which does not phosphorylate glucose. Lactose entry was stimulated 3-fold when cells were also presented with readily metabolizable substrates. Entry of omicron-nitrophenyl-beta-D-galactopyranoside (ONPG) was only slightly elevated (1.5-fold) under the same conditions. The effects of arsenate treatment and anaerobiosis suggest that lactose entry may be limited by the need for reextrusion of protons which enter during H+/sugar cotransport. Entry of omicron-nitrophenyl-beta-D-galactopyranoside is less dependent on the need for proton reextrusion, probably because the stoichiometry of H+/substrate cotransport is greater for lactose than for ONPG.


Two purine nucleoside phosphorylases in Bacillus subtilis. Purification and some properties of the adenosine-specific phosphorylase. Two purine nucleoside phosphorylases (purine-nucleoside:orthophosphate ribosyltransferase, EC 2.4.2.1) were purified from vegetative Bacillus subtilis cells. One enzyme, inosine-guanosine phosphorylase, showed great similarity to the homologous enzyme of Bacillus cereus. It appeared to be a tetramer of molecular weight 95 000. The other enzyme, adenosine phosphorylase, was specific for adenosine and deoxyadenosine. The molecular weight of the native enzyme was 153 000 +/- 10% and the molecular weight of the subunits was 25 500 +/- 5%. This indicates a hexameric structure. The adenosine phosphorylase was inactivated by 10(-3) M p-chloromercuribenzoate and protected against this inactivation by phosphate, adenosine and ribose 1-phosphate.


Purification of pantoate and dimethylmalate dehydrogenase from Pseudomonas fluorescens UK-1. Pantoate dehydrogenase and dimethylmalate dehydrogenase were purified 69- and 112-fold, respectively, from Pseudomonas fluorescens UK-1 by ammonimu sulphate precipitation. Ultrogel AcA 34 gel filtration, hydroxyapatite column chromatography, heat treatment and Ultrogel AcA 44 gel filtration. The enzymes were evaluated for homogeneity (pantoate dehydrogenase was estimated to be about 95% pure) by disc and sodium dodecyl sulphate gel electrophoresis and by immunodiffusion. Pantoate and dimethylmalate dehydrogenases have molecular weights of 83 000 and 138 000, respectively, and are dissociable into four identical subunits with molecular weights of 24 000 and 34 000.


An oviducal enzyme isolated by affinity chromatography which acts upon the vitelline envelope of Bufo arenarum coelomic oocytes. A trypsin-like oviducal proteinase acting upon the vitelline envelope of Bufo arenarum coelomic oocytes has been purified to apparent homogeneity by gel filtration on Sephadex G-200 and by affinity chromatography on a column of Sepharose 4-B containing covalently bound concanavlin A (Con A). The biologically active molecule migrated as a single band of protein upon SDS polyacrylamide gel electrophoresis.


Formation of spirodilactone of 4-(2'-carboxyphenyl)-4,4-dihydroxybutyrate from 2-succinylbenzoate in cell-free extracts of Micrococcus luteus. The enzyme mediated ATP- and, to a lesser extent, CoASH-dependent synthesis of the spirodilactone of 4-(2'-carboxyphenyl)-4,4-dihydroxybutyrate from 2-succinylbenzoate has been demonstrated in membrane-free extracts of Micrococcus luteus and Escherichia coli. The suggestion is made that the spirodilactone is the product of an aberrant reaction involving a compound that is normally an intermediate in the conversion of 2-succinylbenzoate to 1,4-dihydroxy-2-naphthoate.


Isolation and characterization of an enriched Golgi fraction from rat brain. A fraction rich in membranes of the Golgi apparatus was isolated from rat brain by discontinuous density gradient centrifugation. The fraction sedimented at the characteristic Golgi density of 1.11--1.15 (g/cm3, 5 degrees C) and had specific activities of Golgi-marker enzymes (N-acetyllactosaminyl synthetase, glycoprotein (Fetuin) galactosyltransferase, thiamine pyrophosphatase), 6--7 times over those of the original homogenates. The recovery of the enzyme activities in this fraction ranged from 17 to 31 %. The incorporation [3H]fucose into glycoproteins was 3-fold higher than in homogenate. Recovery and relative specific activities of marker enzymes for other subcellular organelles were low. Electron microscopic analysis of the fraction revealed in the presence of Golgi structures, namely, large sacs or plates with attached tubules and "blebbing" of the tubules into the vesicles.


Comparative study of kinetoplast DNA in culture, blood and intracellular forms of Trypanosoma cruzi. A comparative study has been made on the kinetoplast DNA (kDNA) from I in culture epimastigote, blood trypomastigote and intracellular amastigote stages of Trypanosoma cruzi. The basic properties of the kDNA in all 3 forms were identical. Thus the DNA was in the form of networks of density 1.698-9 g/cm3 and with sedimentation coefficients (S20w) of approximately 5500, the networks being composed of large complexes of minicircular and apparently linear molecules, the former having contour lengths of 0.45 MICROMETer. Several differences were noted. The ultrastructural arrangement of the kDNA in the kinetoplast of the blood stage consisted of three to four double rows of DNA as compared to one double layered row in the other two stages. There was proportionately more kDNA in the blood stages, suggesting that, since the networks have apparently the same size (see above), more than one is present. DNA loops situated at the periphery of the kDNA networks were observed in higher proportion in blood and intracellular forms. Dimeric and oligomeric circles were present in the kDNA of the blood and intracellular stages in much greater proportion than in culture epimastigote stages. Few large circular molecules, heterogeneous in size, were also observed in intracellular blood stages. There were some differences, mainly quantitative, in the gel electrophoresis patterns after endonuclease digestion.


Thyrotropin, prolactin and growth hormone response to synthetic thyrotropin-releasing hormone in newborn infants. The effects of 50 microgram synthetic thyrotropin-releasing hormone (TRH) intravenously on thyrotropin (TSH), prolactin (PRL) and growth hormone (GH) levels were studied in 8 normal male newborns during the first hours of life. Mean plasma GH concentrations were similar to baseline values during the period of study; on the contrary, plasma PRL and TSH values increased in all infants after TRH administration. These data demonstrate a normal pituitary reserve of PRL and TSH in the early period of human life.


Prostaglandin-forming cyclooxygenase in renal medulla of spontaneously hypertensive rats during development. Increased activity of prostaglandin synthetase in spontaneously hypertensive rats (SHR) during development has previously been reported. The investigations reported herein utilized an immunofluorescent technique to qualitatively assess the activity of the first enzyme in the PGE2 biosynthetic pathway, the prostaglandin-forming cyclooxygenase (PG-C), in SHR kidneys during development. PG-C antigenicity increased from 4 to 15 weeks in both SHRs and Wistar Kyoto normotensive rats. However, in agreement with our previous in vitro data no difference in PG-C was observed between strains at any age.


Behavioral correlates of social dominance. The behavioral correlates of dominance in the squirrel monkey were explored using a variety of operant tasks. Compared to more submissive animals, more dominant ones showed higher operant rates, more consistent work output, and a higher degree of "frustration tolerance." These data suggest that dominance in the squirrel monkey is not related solely to physical size, but to a variety of behavioral traits which lead to more adjustive problem-solving behavior.


Human lymphoid cells of the intestinal mucosa showing specificity for both immunoglobulin light and heavy chains. The presence of single human lymphoid cells expressing on the one hand both kappa and lambda light chain and on the other hand either both mu and alpha or both mu and gamma heavy chains, was observed in the intestinal mucosa. The variability of the frequency of such cells, both from one subject to another, or in the same sample, is a characteristic of this population.


Comparison of the stability of phycocyanins from thermophilic, mesophilic, psychrophilic and halophilic algae. Protein unfolding of eight different phycocyanins was investigated utilizing circular dichroism and visible spectra. The phycocyanin samples were extracted from algae that are normally found in vastly different environments, and are classified as mesophilic, thermophilic, halophilic and psychrophilic. The ability of these proteins to resist the denaturant urea is in the order of thermophile greater than mesophile, halophile greater than psychrophile. Based on a two-state approximation the apparent free energies of protein unfolding at zero urea denaturant concentration, deltaGH2Oapp, were found to range from 2.4 to 8.8 kcal/mole for the eight phycocyanins at pH 6 and 25 degrees C. The proteins from the thermophile are generally more stable than those from the mesophile. An extra stability of the halophile is believed due to the specific interaction of the proteins and the ions in solution. A correction for deltaGH2Oapp due to minor amino acid differences reveals that the stability and the structural properties of these proteins are primarily affected by this minor difference in amino acid compositions.


Studies on the stability of immobilized xanthine oxidase and urate oxidase. The stability of immobilized preparations of xanthine oxidase and urate oxidase was studied, and optimized, because of the potential joint use of both enzymes in clinical analysis. Xanthine oxidase was immobilized on cellulose, Sepharose, hornblende, Enzacryl-TIO, and porous glass. Thehalf-lives of these preparations at 30 degree C ranged from 40 min to 5.0 hr. In this respect immobilized enzyme resembled soluble enzyme in dilute solution (0.11 mg/ml), when the half-live was about 3.5 hr. More concentrated enzyme solution (1 mg/ml) had a half-life of 64 hr, and was, therefore, considerably more stable than the untreated immobilized xanthine oxidase preparations. Inclusion of albumen in storage and assay buffer increased the half-life of bound xanthine oxidase. So also did treatment with glutaraldehyde: in the case of xanthine oxidase bound to Enzarcyl-TIO such treatment increased the half-life at 30 degree C from 3 hr to about 100 hr. Immobilized xanthine dehydrogenase was more stable than immobilized xanthine oxidase: the dehydrogenase lost no activity during continuous assay for 5 hr at 30 degree C. The stability of immobilized urate oxidase depended on the quantity of enzyme used and on the time of stirring during immobilization: thus a preparation was made (by stirring urate oxidase (48 mg/g support) with Enzacryl-TIO for 24 hr) which lost no activity during 350 hr at 30 degree C.


[Nucleic acids in the tissues of white rats with toxic liver damage following protein starvation and parenteral nitrogen nutrition]. The authors studied the content of nucleic acids in the tissues (the skeletal muscle and myocardium, the liver) in albino rats with toxic affection of the liver caused by ccl4, in protein deficiency and parental administration of amino acid mixture of moriamin S-2 and "improved" caseine hydrolysate. Protein deficiency in albino rats with toxic hepatitis was accompanied by a considerable increase of RNA and DNA in the skeletal muscle and the myocardium with a simultaneous reduction of their level in the liver. The RNA/DNA ratio changed. The RNA content in the hepatocytes diminished on account of reduction of the nuclear RNA fraction closely bound with chromatin. Parenteral administration of nitrogen preparations led to normalization of the nucleic acid content in all the tissues under study.


[Anabolic effectiveness of nitrogenous preparations for parenteral nutrition in the presence of toxic liver damage]. The authors studied the assimilation of nitrogen preparations--moriamin S-2 and "improved" caseine hydrolysate in parenternal administration to 100 albino rats. Healthy animals and those with toxic affection of the liver induced with CCl4 were experimented upon. In healthy animals administration of nitrogen preparations led to the change of negative nitrogen balance into a positive one, normalized the content of blood and tissue amine nitrogen deranged in protein deficiency. Assimilation of nitrogen preparations fell considerably in toxic hepatitis. An 8-day parenteral nutrition failed to change the negative nitrogen balance into positive, and did not eliminate hypoproteinemia; however, it normalized the amine nitrogen concentration in the blood and tissues.


[Development of allergic reactivity to Artemesia pollen during combined sensitization to pollen and microbes]. Some regularities of formation of hypersensitivity of the immediate type to the pollen of Artemisia absinthium were studied under conditions of combined hypersensitivity to pollen and Brucella abortus 19-BA vaccine strain; the latter was administered 3, 12, and 28 days after the pollen. The degree of specific allergic reconstruction to the pollen was studied by passive skin anaphylaxis after Ovary, indirect degranulation of mast cells of healthy rats, and by general anaphylaxis in response to intravenous injection of the Artemisia absinthium pollen water-salt extract. Early formation of allergy to the pollen was observed in the groups of animals with combined hypersensitivity to the pollen and brucellae. The degree of allergic reactivity to the pollen allergen was more expressed in the groups with combined allergy than in those with pure pollen hypersensitivity at all the stages of this experiment.


Microscopic method for assaying F cell production: illustrative changes during infancy and in aplastic anemia. Fetal hemoglobin (HbF)-bearing reticulocytes (F reticulocytes) can be detected in peripheral blood by a modification of the microscopic single-cell radial immunodiffusion method. Thereby otherwise inappreciable changes in HbF production can readily be recognized. F reticulocyte frequencies are reporducibly measurable whenever the product of whole blood HbF-bearing red cell (F cell) frequency and reticulocyte frequency is approximately 5 X 10(-4) or greater. Serial analyses of F reticulocytes and nonreticulocyte F cells (F erythrocytes) illustrate that (1) levels of F reticulocytes and F erythrocytes are persistently similar in normal adults with more than 6% F cells and thus cell survival times of F and non-F cells must be essentially the same, (2) changing levels of F reticulocytes can be sensitive predictors of later changes in mature F cell frequencies during infancy and in adults recovering from aplastic anemia, and (3) alterations in F reticulocyte frequency and the amount of HbF per F reticulocyte are discordant in some settings but concordant in others.


Heterogeneity of human whole blood platelet subpopulations. II. Use of a subhuman primate model to analyze the relationship between density and platelet age. A subhuman primate model was developed to ascertain whether or not platelet heterogeneity could be explained by aging in the peripheral circulation. Density-dependent platelet cohorts, postulated to represent cells of different ages, were isolated on isosmolar arabinogalactan gradients and labeled with radiochromium. Mean platelet lifespan was measured for the different density cohorts, and simultaneous sequential density distribution analysis was performed to follow changes in cell density during aging. The average mean lifespan of light platelets was 74.6 hr, compared to 313.6 hr for heavy platelets. After injection, labeled light platelets were recovered only in the gradient light region, in contrast to labeled heavy platelets, which were initially restricted to the dense region and progressively migrated to the light region during their lifespan. This study supports the hypothesis that platelet age in unstressed primates correlates with cell density and provides a rationale for the use of "age-dependent" markers to estimate platelet turnover rates.


Is childhood asthma being underdiagnosed and undertreated? Thirty-four cases of asthma in children referred to outpatient clinics in Newcastle upon Tyne (16 cases) and London (18 cases) were reviewed. In both cities there was evidence of inappropriate diagnosis and treatment by general practitioners. One of the main factors seemed to be doctors' reluctance to use the word "asthma," even when a history of episodic wheezing strongly suggested the diagnosis. Freer use of the word "asthma" might help parents to co-operate in managing asthmatic children and allow them to be better prepared to cope with severe asthmatic attacks if they occur.


Prenatal removal of frontal association cortex in the fetal rhesus monkey: anatomical and functional consequences in postnatal life. Anatomical and behavioral consequences were examined following bilateral or unilateral resections of the presumptive dorsolateral prefrontal cortex in 4 rhesus monkey fetuses at E102 (102nd embryonic day), E104, E106 and E119, with subsequent replacement in utero and delivery at later stages of development. Unoperated monkeys of appropriate pre- and postnatal ages served as controls for certain features of morphogenesis and tectogenesis. Monkeys given comparable unilateral or bilateral lesions at selected postnatal ages were used as controls for the behavioral effects of prenatal resection as well as for neuropathological examination. The behavioral evaluation provided evidence for extraordinary preservation of function after frontal cortical resection in the prenatal period. During the entire course of its postnatal development, a monkey that sustained bilateral resection as a fetus at E106 performed surprisingly well and clearly with the competence of unoperated monkeys of identical or older ages on tasks selective for dorsolateral prefrontal cortical function; it did not exhibit deficits characteristic of animals that have comparable or even smaller resections as juveniles or adults. Following either unilateral or bilateral resection prior to E106, ectopic sulci and gyri appeared in intact regions of the frontal lobe of each hemisphere, as well as in locations at a considerable distance from the lesions, such as the temporal and occipital lobes. The cytoarchitectonic characteristics of the cortex in these anomalous gyri were typical of adjacent cortex in the regions in which they appeared. Examination of the parvocellular subdivision of the mediodorsal thalamic nucleus yielded the novel finding that this principal source of essential projections to the dorsolateral prefrontal cortex contained a virtually normal number of neurons in postnatal monkeys, following unilateral or bilateral lesions performed at or before E106, and exhibited only a partial loss of neurons after surgery at E119. The same cells regularly degenerate following comparable resections in the postnatal period. The survival of these neurons in prenatally operated monkeys is unprecedented, and can be explained by several hypotheses, including the possibility that their axons become rerouted to their structures in the absence of their normal targets. The present results thus demonstrate that ablation of a circumscript area of the frontal cortex in a non-human primate a full two months before birth may leave the functions later to be subserved by that area unaffected. Such early surgical intervention results in significant alteration in the gross morphological characteristics of the cortex, and apparently induces a functionally compensatory reorganization of thalamocortical connections. Similar principles may account for sparing of function following early damage to the human brain.


Selective amplitude histograms: a statistical approach to EEG-single unit relationships in generalized epilepsy. Statistical analysis techniques that permit detection and quantification of EEG-single unit correlations were employed in a study of generalized penicillin epilepsy in the cat. Single unit activity was recorded in cerebral and cerebellar cortices and compared to the locally recorded EEG. It was found that during generalized paroxysmal activity the background EEG was significantly related to the time of occurrence of unit discharge. The degree of synchrony of various units differed but tended to increase as the duration and extent of paroxysmal activity increased. These relationships were usually not evident upon visual inspection of the original unprocessed data but were consistently detected by the analysis techniques described here. The results demonstrate the usefulness of a statistical approach to the analysis of single unit data and suggest that, not only is there a significant relationship between paroxysmal events and neuronal activity in generalized penicillin epilepsy, but also an overall trend to closer synchronization of EEG and single unit discharges during nonparoxysmal periods.


The development of hepatic drug-metabolizing enzymes in perinatal guinea pigs: a biochemical and morphological study. The development of four functionally diverse, hepatic enzymes (p-nitroanisole O-demethylase, aniline hydroxylase, carboxyleterase, and glucuronyltransferase (with alpha-naphthol as the aglycone acceptor)) was studied in perinatal Hartley guinea pigs from 8 days prepartum to 28 days postpartum. A good correlation was observed between the activities measured in resuspended Ca2+-aggregated microsomes and the quantities of hepatic smooth endoplasmic reticulum visible by electron microscopic examination at the different stages of development. The study demonstrated that, postnatally, the guinea pig developed competent enzymatic systems as rapidly as did other laboratory species but that, prenatally, these same enzyme(s) systems were much further advanced than those in other species.


Subcellular distribution of lysophosphatidylinositol and lysophosphatidylcholine acyltransferases in rat brain. Lysophosphatidylinositol acyltransferase utilizing arachidonoyl CoA and lysophosphatidylcholine acyltransferase utilizing linoleoyl CoA were measured in subcellular fractions of rat brain. In general, the distribution of activities paralleled that of NADPH--cytochrome c reductase. It is concluded that the endoplasmic reticulum is the major site of these acyltransferase activities in rat brain.


A Zoogloea sp. associated with blooms of Anabaena flos-aquae. Bacteria were found attached to the heterocysts of Aphanizomenon flos-aquae and embedded within the mucilage of both anabaena flos-aquae and Microcystis aeruginosa in freshwater plankton. Electron microscopy of thin sections preceding the peak of an Anabaena flos-aquae bloom showed that the density of bacterial cells was 7.4 X 10(5) cells/ml in the planktonic macroenvironment and 2.6 X 10(11) cells/ml within the microenvironment of cyanobacterial mucilage. The bacteria occurred in aggregates and isolation required that these be dispersed by homogenizing at 50 000 rpm with glass beads. This procedure yielded a single bacterial isolate from blooms of Anabaena flos-aquae during 2 consecutive years. The isolate was flagellated, catalase- and oxidase-positive. Gram-negative, and rod-shaped to pleomorphic. Observation that the isolate required a pH greater than 8 for consistent growth, could not grow alone on liquid media but could grow alone on the corresponding solid media, could grow in liquid media only in the presence of Anabaena, formed tough mucilagenous colonies on solid media only in the presence of Anabaena extract, and rapidly assimilated but did not respire extracellular 14C-labelled organic matter produced by Anabaena suggested that the occurrence of the bacterium in cyanobacterial mucilage was not coincidental but reflected an obligatory bacterial requirement for the biological or physicochemical microenvironment of the mucilage. The bacterial isolate occurred in three growth forms. Either as a planktonic swarmer cell (which showed a positive chemotactic response to the cyanobacterium) embedded in cyanobacterial mucilage, or embedded in its own mucilage derived, in part, from a low molecular weight (below 1300) carbohydrate secreted by the cyanobacterium. These cultural, biochemical, and ecological characteristics suggest that the isolate is a new species in the genus Zoogloea and of potential importance in phytoplankton ecology.


Nitrogen-fixing communities in an intertidal ecosystem. Nitrogen gixation (acetylene reduction) associated with various communities in the intertidal sand and mud flat was measured in situ. Areas which were colonized by algae, plants, and animals generally had significantly higher activities than areas which were visibly uncolonized. The highest activities were measured at sites colonized by a mixed bloom of Oscillatoria-Euglena-photosynthetic bacteria. These areas occupied only about 1% of the surface of the sediments, yet contributed nearly 50% of the biologically fixed nitrogen. Enteromorpha communities also exhibited relatively high activities. Sites were grouped according to the statistical significance of the mean of their activities: group I: uncolonized sand, mud and gravel flats, Eelgrass (Zostera) communities, and snail (amphibola) beds, with an estimated contribution of 1 g N ha-1 day-1; group II: Cockel (Chione) beds, 3 g N ha-1 day-1; group III: Salicornia, Ulva, Juncus, and Enteromorpha, 10 g N ha-1 day-1; and group IV: Oscillatoria-Euglena-photosynthetic bacteria association, 200 g N ha-1 day-1. Enteromorpha and Oscillatoria-Euglena-photosynthetic bacteria communities showed significantly lower rates of nitrogen fixation in the dark than in the light.


Cellular immunity and nutrition in refractory disseminated blastomycosis. In a previously healthy 13-year-old girl with disseminated blastomycosis, immunodeficiency was considered because of lymphopenia and the slow response of her lung disease to therapy with amphotericin B. Cellular immunity was found to be profoundly impaired, with absent delayed cutaneous hypersensitivity to several common antigens, a decreased count of thymus-dependent lymphocytes in the peripheral blood and a greatly diminished in-vitro proliferative response of lymphocytes to phytohemagglutinin (PHA). Humoral immunity was intact. Two additional types of therapy were assessed: subcutaneous injection of transfer factor was associated with an unsustained increase in lymphocyte counts and a positive cutaneous response to PHA but no clinical change; parenteral alimentation to ensure an adequate energy intake was associated with rapid clinical improvement, the development of delayed hypersensitivity to four additional antigens, and the return of lymphocyte counts and proliferative response to normal. These findings suggest that increased energy intake rather than transfer factor therapy was responsible for the child's recovery, and they emphasize the importance of adequate nutrition in the maintenance of intact cellular immunity.


Comparison of silver sulfadiazine and gentamicin for topical prophylaxis against burn wound sepsis. Daily prophylactic application of either 1.0% silver sulfadiazine cream or 0.1% gentamicin cream was compared for effectiveness in preventing bacterial colonization of burn wounds and sepsis. Pseudomonas aeruginosa colonized the wounds of 37% of the 38 patients treated with silver sulfadiazine and 30% of the 33 patients treated with gentamicin; gentamicin-resistant P. aeruginosa colonized the wounds of 21% of the patients treated with gentamicin. Staphylococcus aureus colonization occurred in 55% of the patients treated with silver sulfadiazine, whereas colonization with Candida species occurred in 58% of the patients treated with gentamicin. Although gentamicin-resistant organisms caused no deaths their repeated appearance resulted in discontinuation of prophylaxiz with gentamicin cream. The next year P. aeruginosa strains resistant to gentamicin were isolated from burn wounds of only two patients who had not previously received parenteral therapy with gentamicin or tobramycin. Gentamicin cream should be reserved for treating patients with wounds infected by gentamicin-sensitive P. aeruginosa and those allergic to sulfa drugs. For most patients with burn wounds silver sulfadiazine is safe and effective as an antibacterial agent for topical prophylaxis.


Current management of unstable angina.

Current management of unstable angina. The patient with unstable angina (angina of recent onset, of changing pattern or occurring at rest) is at high risk of myocardial infarction and sudden death. Patients with simple angina of recent onset can generally be managed out of hospital. Those with progressive angina or angina at rest should be admitted to a coronary care unit, kept at bed-rest, and given propranolol and long-acting nitrates when such therapy is indicated. With these approaches the rate of infarction within 1 to 3 months after the onset of unstable angina is about 12% (as compared with 40% before 1970); the mortality in the same period is less than 2% (as compared with 17% before 1970), though during the first year it is about 17%, much higher than in patients with stable angina and in survivors of acute myocardial infarction.Urgent aortocoronary bypass grafting has proven to be unnecessary and probably undesirable for most patients with unstable angina, and is now generally reserved for patients who continue to have angina in hospital while receiving full medical therapy. The ongoing management of patients whose angina is controlled during the acute phase remains controversial. The main options are to operate on every possible patient, to operate only on those with certain distributions of coronary artery lesions, and to operate only on those who have recurrent symptoms. Further studies are required to delineate the etiology and the Optimal management of unstable angina.


Combined modality therapy of Hodgkin's disease: a report on the Stanford Trials. A total of 440 previously untreated patients with Hodgkin's disease have been treated on randomized clinical trials at Stanford University, testing the value of combined modality therapy. A group of 244 patients with stages I, II and III were treated between 1968 with radiotherapy alone or combined with adjuvant MOPP chemotherapy. The adjuvant MOPP significantly improves the initial freedom from relapse (FFR) duration but improvement in survival is only minimal and not yet significant. The ability to induce a second durable remission after initial treatment failure results in freedom from second relapse rates (FF2dR) which more closely parallel survival figures than FFR. Adjuvant MOPP cannot yet be recommended as a routine adjuvant in the radiation maanagement of Hodgkin's disease. A pilot trial of the role of radiation therapy in the chemotherapy management of stage IV patients does not indicate an advantage of the irradiation. Preliminary analyses of new treatment programs in 163 patients with all stages of disease treated between 1974--1977 indicate improved survival and FFR rates, the majority of patients receiving combined modality therapy. Only three patients have died and nine patients have relapsed during the three year period of these new trials.


Uridine 5'-diphosphate galactose: glycoprotein galactosyl transferase activity in exfoliated bladder epithelial cells in rats fed N-(4-(5-nitro-2-furyl)-2-thiazolyl) formamide. Urine samples of normal male Fischer rats or rats fed 0.2% N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide for 6,8 or 30 weeks were collected and centrifuged 50 weeks after beginning treatment. After being sonicated and assayed (with purified desialylated ovine submaxillary mucin as acceptor glycoprotein), the exfoliated bladder cells obtained from the urines of treated rats showed uridine 5'-diphosphate galactose:glycoprotein transferase activity. The specific enzymatic activity of the enzyme from cells of 30-week-treated rats was about 10 times higher than from normal rats. The enzyme from cells of hyperplastic rats (treated 6 or 8 weeks) was only slightly higher in specific activity than that of normal rats. A similar was obtained at a later stage of bladder tumor induction, when the urines from 30-week-treated rats contained blood. A correction was made for protein contributed by the blood clot. The possibility that the blood clot contributed galactosyl transferase activity was excluded. Activity of the enzyme was detected in normal rat bladder tissue and in normal human urine.


Tissue-mediated mutagenicity of vinylidene chloride in Salmonella typhimurium TA1535. Vinylidene chloride is weakly positive in the Salmonella typhimurium TA1535 test, mediated by kidney and liver post-mitochondrial supernatant (S-9 mix) from normal mice, but strongly positive with the S-9 mix from the induced animals. In the case of mediation by rat tissue, only liver S-9 mix from induced animals affords a significant positive response. These findings agree with the greater availability in treated mice than in rats of reactive vinylidene chloride metabolites, 1,1-dichloroethylene oxide and chloroacetyl chloride [5], and with the vinylidene carcinogeneicity found in mice but not in rats [9]. Exploratory tissue-mediated testing of vinylidene chloride involving liver S-9 mix from marmosets and man suggests a trend in the generation of alkylating metabolites and their reactions with bacterial DNA for these primates which resembles rats more than mice.


Effects of selenium and retinoic acid on the metabolism of N-acetylaminofluorene and N-hydroxyacetylamino-fluorene. Male albino rats were given 4 ppm selenium (Se(as Na2SeO3) in the water, or 0.25% retinoic acid was added to the basal diet for 3 days. Control and treated animals were given 17 mg acetylaminofluorene (AAF)-9-14C or N-hydroxyacetylaminofluorene (NOHAAF)-9-14C/kg body wt. The administration of Se enhanced glucuronyl transferase activity by 100% and inhibited p-nitrophenol-sulfotransferase by 50%. Retinoic acid enhanced the glucuronyl transferase 37% and inhibited the p-nitrophenol-sulfotransferase by 50%. Pretreatment with Se lowered levels of both AAF and NOHAAF in liver tissue by 30%. Se administration lowered the binding of the labeled carcinogens, or metabolites thereof, to the liver DNA and tRNA.


Formation of glucuronic acid conjugates of 7,12-dimethylbenz(a)anthracene phenols in 7,12-dimethylbenz(a)anthracene-treated hamster embryo cell cultures. Secondary cultures of hamster embryo cells exposed to 0.5 nmol [G-3H]7,12-dimethylbenz(a)anthracene (DMBA) per ml medium metabolized more than 90% of the DMBA within 48 hr. Samples of medium were extracted with chloroform, methanol, and water. The chloroform phases contained about one-third of the DMBA metabolites; the major chloroform-extractable metabolite was 8,9-dihydro-8,9-dihydroxy-7,12-dimethylbenz(a)anthracene. Beta-glucuronidase treatment of the aqueous methanol-soluble metabolites converted almost one-half of them to chloroform-soluble metabolites, of which more than 80% were identified as phenolic derivatives of DMBA. Similar metabolite profiles were obtained by treating the medium with beta-glucuronidase before chloroform extraction. Separation of the methyl group-hydroxylated derivatives of DMBA from the phenolic derivatives was accomplished by high-pressure liquid chromatography. Small amounts of hydroxymethyl derivatives were detected only in the chloroform-extractable material, whereas DMBA phenols were the major component of the beta-glucuronidase-released mateirla. These results indicate that the major pathway of DMBA metabolism in hamster embryo cells is oxidation of the aromatic rings and not oxidation of the methyl groups.


Concurrent depression of tumor macrophage infiltration and systemic inflammation by progressive cancer growth. Macrophage accumulation during the growth of a peritoneal ascites and three s.c. tumors in two animal species was analyzed and correlated with the capacity of the same tumor-bearing host to respond to inflammatory stimuli at sites distant to the tumor. Two of the three s.c. tumors induced systemic defects in macrophage accumulation; the tumor that did not (P-815 mastocytoma) did depress inflammation when transplanted to the peritoneal site. Macrophage accumulation within different tumors varied but, for a given tumor, it occurred in proportion to tumor growth when systemic inflammatory reactions were normal. However, the tumor to macrophage ratio increased dramatically and concurrently with onset of the generalized defect in macrophage inflammatory responsiveness. Accordingly, we concluded that macrophage mobility tested at remote sites is indicative of inflammatory events within the tumor. However, the antiinflammatory effect directed against macrophages is probably not a significant factor in tumor emergence since the required number of tumor cells was large and variable between not only tumors but also sites of transplantation.


Protection against chemotherapy toxicity by IV hyperalimentation. A prospective randomized trial was conducted comparing the addition of iv hyperalimentation (IVH) to Corynebacterium parvum, isophosphamide, and adriamycin (CIA) chemoimmunotherapy in 26 patients with extensive squamous cell lung cancer. Thirteen patients were entered in each treatment arm of the study and IVH was administered before and after the first course of CIA for a total of 31 days. The major dose-limiting toxic effect of CIA was leukopenia. Less myelosuppression was observed for the patients receiving IVH. The difference in the lowest recorded leukocyte and neutrophil counts between the two groups was significant (P = 0.03 and 0.01, respectively). Also, a significant decrease (P = 0.06) in nausea and vomiting associated with chemotherapy administration was found for the IVH gorup. The differences in toxic effects between each group were not maintained over subsequent courses of therapy when both groups received CIA alone. The prevention of the toxic effects of chemotherapy by IVH suggests a means of giving higher chemotherapy doses with the intent of increasing tumor response and patient survival.


A protein released by DNAase I digestion of drosophila nuclei is preferentially associated with puffs. Antisera have been produced against five molecular weight subfractions of the Drosophila proteins readily extracted from nuclei following limited DNAase I digestion. Immunofluorescence staining techniques were used to assess the distributions of these proteins in the polytene chromosomes of Drosophila. In three cases, the antigens were widely distributed; in one case, the antigens appeared to be slightly more concentrated at active loci; and in one case, the antigens were strongly concentrated at a defined set of loci, including puffs and most of the loci which are active (puffed) at some time during third instar larval and prepupal development. The latter distribution pattern differs from that of RNA polymerase. Nonhistone chromosomal proteins of this type may have a key role in establishing and/or maintaining the altered chromatin structure characteristic of the active state.


A novel arrangement of tandemly repeated genes at a major heat shock site in D. melanogaster. Three cloned segments of Drosophila melanogaster DNA have been isolated that derive from the major heat shock site at 87C in chromosome 3. Each of these segments contains sequences homologous to a class of polysomal poly(A)+ RNAs whose synthesis is induced by heat shock of cultured cells. A combination of R loop, heteroduplex and restriction fragment maps of these segments reveals that their RNA-homologous sequences are arranged in tandemly repeated units, each unit consisting of an alpha element (0.49 kb) joined to a beta element (1.10 kb). The polysomal RNAs homologous to these alphabeta units (1.59 kb) are distributed into three size classes exhibiting approximate lengths of 1, 2 and 3 kb. R loop mapping demonstrated that the sequence of the 2 kb RNA is alphabetaalpha, indicating that it, and presumably the 3 kb RNA, derive from transcripts covering more than one repeated unit. One of the cloned segments contains a variant repeat unit, alphagamma, located between two alphabeta units. This unit has the same alpha element, but the beta element has been replaced by a nonhomologous gamma element (0.87 kb). Analysis of the total D. melanogaster DNA indicates that the 87C locus contains at least 21 tandemly repeated units, twelve of which were identified as alphabeta units and six as alphagamma units. The 21 or more units are distributed among at least three different tandem arrays separated by different spacer regions, one of which is within a cloned segment. Sequences in the gamma element, but not those in the alpha or beta elements, are also found at 87A, which contains another heat shock site. The possible roles of the alphabeta-type RNAs and of the gamma sequences are discussed in the light of these results.


Two hybrid plasmids with D. melanogaster DNA sequences complementary to mRNA coding for the major heat shock protein. The isolation and partial characterization of two cloned segments of Drosophila melanogaster DNA containing "heat shock" gene sequences is described. We have inserted sheared embryonic D. melanogaster DNA by the poly(dA-dt) connector method (Lobban and Kaiser, 1973) into the R1 restriction site of the ampicillin-resistant plasmid pSF2124 (So, Gill and Falkow, 1975). A collection of independent hybrid plasmids was screened by colony hybridization (Grunstein and Hogness, 1975) for sequences complementary to in vitro labeled polysomal poly(A)+ heat shock RNA. Two clones were identified which contain sequences complementary to a heat shock mRNA species that directs the in vitro synthesis of the 70,000 dalton heat-induced polypeptide. Both cloned segments hybridize in situ to the heat-induced puff sites located at 87A and 87C of the salivary gland polytene chromosomes.


RNA synthesis and control of cell division in the yeast S. cerevisiae. Cells of the yeast Saccharomyces cerevisiae rapidly accumulated in the G1 phase of the cell cycle when exposed to the chelating agents o-phenanthroline (OP) or 8-hydroxyquinoline (HQ). Zinc salts fully reversed the growth-inhibitory effect of both OP and HQ. Cells treated with these chelating agents showed limited RNA accumulation and little RNA degradation. Rates of RNA synthesis were drastically reduced by low concentrations of these compounds. Whereas rates of protein synthesis were essentially unaffected. Rates of synthesis of mRNA and tRNA were less affected than were rates of synthesis of high molecular weight RNA. Processing of ribosomal precursor RNA was altered. these results suggest that the primary effect of OP and HQ is on rRNA synthesis. RNA metabolism must therefore have a key role in the regulation of the cell cycle.


Reversal by aminoguanidine of the inhibition of proliferation of human fibroblasts by spermidine and spermine. The inhibitory effect of the polyamines, spermidine and spermine, on the proliferation of human fibroblasts in culture was found to be reversed by the addition of aminoguanidine (AM), a specific and highly effective inhibitor of diamine oxidase (DAO) present in fetal calf serum (FCS). Aminoguanidine itself in concentration as high as 10(-3) M exhibited no effect upon cell proliferation nor did putrescine at similar concentrations. However, at higher concentrations of putrescine, cell proliferation was inhibited and this inhibition was unaffected by the addition of mM concentrations of AM. These studies support earlier hypotheses on the mechanisms of the toxic effects of polyamines on cell proliferation and establish further that the diamine oxidase-catalyzed metabolism of spermine and spermidine is necessary for their toxic effects in cell culture.


[Electron microscope observation of "thyroidectomy cells" in the hypophyseal ventral lobe of a Chondrichthyan fish, the small spotted dogfish, Scyllium canicula Cuv]. In the adenohypophysis of very young spotted dogfish, only one kind of granulated cells show strcutural changes following thyroidectomy: the small granule cells of the ventral lobe (granule diameter about 110 nm). These cells are similar to the thyrotrophs of other Vertebrates, and well distinct from the supposed gonadotrophs found in the same lobe.


[Variation of the inflammatory response of an autologus implant function and viability: mixed and unilateral inflammatory response]. The behaviour of an autologous grafted implant depends upon its viability. In our study, pericardial implants were used as cardiac valves. Results showed that when using fresh live pericardium, an inflammatory response developed destroying it within 8 days. When implants cells were killed without alteration of their antigenicity--in order to prevent rejection--cellular growth and neovascularisation from implantation site occurred leading to a secondary live graft. Controlled killing of the implant seems to be an essential step in order to get live grafts.


[Radioautographic identification of central monoaminergic neurons by local micro-instillation of tritiated serotonin or noradrenaline in cats]. Monoaminergic neurons in nuclei raphe dorsalis and locus coeruleus of the Cat may be visualized by radioautography after local micro-instillation of tritiated serotonin and noradrenaline. The concomitant administration of the appropriate tracer with the other biogenic amine in non radioactive form permits a specific identification of serotoninergic and catecholaminergic nerve cell bodies. A small contingent of presumptive serotoninergic neurons is thus demonstrated in the region of the locus coeruleus.


[Comparison of estradiol benzoate and dihydrotestosterone propionate with testosterone propionate on the induction of male-like sexual behavior in female sheep ovariectomized as adults]. Daily I/M injections of 200 microgram of oestradiol benzoate induce in female Sheep, ovariectomized as adults, male-like sexual reactions similar to what is obtained with 10 mg testosterone propionate per day. Simultaneously, a permanent female sexual receptivity is observed. Oestradiol benzoate at the dose of 20 microgram/day, and dihydrotestosterone propionate (10 mg/day) are both totally ineffective. The result support the hypothesis of the necessity of the aromatization of testosterone for the action on sexual behaviour.


[The problem of calculating the circular dichroism signals of the 10 nearest neighbor configurations of DNA]. Due to linear constraints over the nearest neighbour frequencies the order of the linear dependence of DNA CD spectra is 8. The computation of the CD signals of the 10 nearest neighbour configurations is therefore not possible if these signals are linearly independent. We show that the method proposed by Allen et coll. does not allow to obtain these signals.


[Modes of tetracycline resistance in Bacillus thuringiensis]. The sensitive strain and tetracycline-resistant mutants exhibit two types of growth curves after addition of tetracycline. The level of resistance of these strains is enhanced by a short period of preincubation with a non-inhibitive concentration of tetracycline.


[T-Lymphocyte regulation of contact sensitivity: effect of thymectomy in adult mice]. 6 weeks after adult thymectomy (ATx) in the Mouse, the contact sensitivity reaction to dinitrofluorobenzene (DNFB) is enhanced. 4 to 7 months after ATx, this reaction is deeply, but incompletely depressed, whereas the concomitant antibody response is not affected. These results suggest that both a suppressor, and an amplifier, T lymphocytes, the life span of which is different after ATx, are involved in the regulation of contact sensitivity. The effect of circulating thymic factor on this reaction suggests that this factor acts exclusively, at least in short treatments, on the suppressor function.


[Effects in man of a total adrenal cortical extract on delayed immunity and lymphocyte sub-populations]. After a 10 day treatment with total adreno-cortical extract, administered to 11 subjects, the following results were obtained: increase of delayed cutaneous responses to various antigens, increase in the number of null lymphocytes, decrease in the number of B lymphocytes. These changes did not occur in 16 control patients who were not treated.


[Inhibitory effect of the tissue antagonism of interferon on the development of 180/TG Crocker tumor: recognition of a new murine lectin]. Crude and purified murine lectin preparations are extracted from costal cartilage (TAI). They inhibit the antiviral state induced by interferon. They also agglutinate the Crocker 180/TG tumor cells. After IP inoculation in mice, the purified lectin preparation significantly decreases tumor incidence and increases the animal's life span.


[Effects of testosterone propionate on LH and testosterone concentrations before and after the injection of gonadoliberin in young bulls pre-treated with dexamethasone]. This preliminary study has been performed on four young twin bulls, one pair aged 6 months and the other one aged 12 months. Plasma LH and testosterone were assayed from frequent blood sampling. Testosterone propionate (TP) following dexamethasone treatment did not significantly influence the LH response to gonadoliberin compared to the twin controls, although testosterone concentrations were high in those TP treated animals.


[Study of gangliosides in normal (C57B1/6J) and quaking mice. Presence and characterization of a labile ganglioside in alkaline solution]. In the adult Quaking mutant, there is an increase in some polysialogangliosides (GT1 and Gq)  and a 50% decrease in BM1 content. Thus, the latter ganglioside, which is a constituent of mature myelin, could be linked to one of the late steps of the myelination process. A novel ganglioside ALG1, which appears during the postnatal development of brain, has been isolated and purified; it  contains an alkali labile linkage and after alkaline treatment yields GT1b. Comparison between normal Mice and Quaking mutants does not show any significant quantitative differences, under our conditions. At the stage at which it appears, it could be linked to steps of brain maturation other than myelimation.


[Sensitivity of crayfish muscle fibers to glutamate and to natural transmitter in low calcium solutions]. The response of crayfish muscle fibres to glutamate, a putative transmitter at its excitatory motor synapses, decreases when extracellular calcium is reduced, whereas the sensitivity to natural transmitter s not modified. Three possible mechanisms are proposed.


[Thixotropic effect and electro-osmosis as possible factors of membrane electrical excitability]. Cation motion under the electrical field might break the membrane structure. In the temporary wake of each cation conductivity would be increased. Thus a zone of negative resistance would occur in the intensity-voltage relation. Membrane dehydration, due to electro-osmosis, might induce repeated flip-flops of current at the limits of the negative resistance zone.


[Purification of 2 succinic semi-aldehyde reductases from the human brain]. Two succinic semi-aldehyde reductases (A and B) have been purified to electrophoretic homogeneity. Enzyme A, which is a monomer of molecular weight 40,000 +/- 5,000 reduces a wide range of aldehydes and is inhibited by some barbiturates and certain anticonvulsants. Enzyme B, which is a dimer of molecular weight 80,000 +/- 10,000, is very specific for succinic semi-aldehyde and is not inhibited by the aforementioned compounds.


[Convulsions induced by enanthotoxin in the rat: correlation between electrophysiological modifications and clinical signs]. When injected intraperitoneally in the Rat, oenanthotoxin induces localised myoclonic jerks and generalised tonic-clonic seizures. Electrocorticographic recordings reveal typical epileptic spikes. Correlations between electromyographic bursts, electrocorticographic events and subcortical activities support the hypothesis that the clonic jerks and the generalised seizures have a cortical orgin.


[Disappearance of the gliofibrillar protein (GFA) during the cultivation of glioblastoma cells]. Absence of GFA in established cell lines from glioblastomas promped us to look for this protein from the first replication of malignant cells in vitro and in following subcultrues. Our results show that GFA disappeared during the first twelve replications, often before the fifth replication. No correlation was observed with a morphological transformation of the cells.


A comparative study of three approaches to the routine quantitation of human serum proteins. The Laser Nephelometer PDQTM (Hyland Division, Travenol Laboratories Inc.) and the Abbott Bichromatic Analyser 100 (Abbott Laboratories) were compared with a radial immunodiffusion method. Seventy-eight serum specimens collected during routine blood testing were aliquoted and quantitated by the three procedures described. The nephelometric system was used as described by Hyland in their instruction accompanying the LAS-R Nephelometric test kits. The ABA-100 was used with a filter unit transmitting at 340 and 650 nm and at a water bath temperature of 30 degrees C. The Laser Nephelometer correlated well with the RID system giving a correlation coefficient varying from 0.94 for IgA to 0.79 for C3. It was not possible to quantitate IgA by the turbidometric method using the ABA-100. Results obtained for the other proteins were satisfactory and the correlation coefficient with the RID method varied from 0.88 for IgG to 0.90 for C3. The RID procedure in routine use takes 5 hours technologist time and a 16-hour incubation period to produce results for IgG, IgA and IgM on nine patient specimens. Using the Laser Nephelometer to obtain the same results on 20 patient specimens took two to three technologist hours. Nephelometry, therefore, appears to be a satisfactory alternative to RID with a comparable precision and a great saving in technologist time.


Improved enzymatic assay of chloramphenicol. We partly purified R-factor-encoded chloramphenicol acetyltransferase (EC 2.3.1.28) from a highly choloramphenicol-resistant mutant derived from Escherichia coli W677/R5. The preparation permitted rapid quanitation of chloramphenicol by use of [14C]acetylcoenzyme A, removing the diacetylated product by selective adsorption onto micropore filters. Succinyl and glucuronyl 3-hydroxyl esters of chloramphenicol were not active as substrates for the preparation, nor were they active as inhibitors. The enzyme was free of chloramphenicol reductase activity, and utilizes other biologically active chloramphenicol analogs. Other antibiotics, at concentrations commonly found in human sera, and blood preservatives, at concentrations 10-fold that found in blood-collection tubes, did not interfere with the enzymic quantitation of chloramphenicol. We conclude that this enzyme preparation permits rapid clinical quantitation of chloramphenicol.


Double-antibody enzyme immunoassay for nortriptyline. beta-D-Galactosidase (EC 3.2.1.23) from Escherichia coli was conjugated to desmethylnortriptyline by means of a bifunctional cross-linking reagent, dimethyl adipimidate, and used in a double-antibody immunoassay for nortriptyline. Eighty percent of the enzyme activity was retained after conjugation; 75% of the enzyme was conjugated to desmethylnortriptyline. In the final immunoassay the enzyme activity of the bound fraction was determined with o-nitrophenyl-beta-D-galactopyranoside as substrate. The sensitivity, precision, and simplicity of the enzyme immunoassay compared favorable with that of a published radioimmunoassay method. Results for nortriptyline in plasma samples correlated well with those determined by either radioimmunoassay or gas-chromatography.


Triiodothyronine uptake test in which antibody-coated tubes are used compared to two other tests in which precipitation separations are used. A triiodothyronine uptake test in which antibody-coated tubes are used (Clinical Assays) was compared to two other such tests in which silicate tablets (Nuclear-Medical Laboratories, Inc.) or macroaggregated albumin (Diagnostic Corporation of America) are used to separate the bound from the free fraction. The Clinical Assays test showed good linear correlation with the values obtained by the other two tests on 67 samples. The interassay coefficient of variation was 4.8%. The correlation between the values obtained and the clinical evaluation on 60 patients also was essentially the same as for the other two tests, 80%.


The course of facial nerve innervation for the levator veli palatini muscle. The present study was designed to determine the motor nerve pathway of the facial nerve to the levator veli palatini muscle. The experiments were carried out on 10 anesthetized rhesus monkeys. Recorded and analyzed were evoked EMG responses of the levator veli palatini and the orbicularis oris muscles by electrical stimulation to both the facial nerve and its branch within the cranium. Muscle action potentials (M-waves) from the two muscles could be recognized on stimulating the facial nerve at the petrosal area of the temporal bone. On stimulating the greater petrosal nerve, M-waves from the levator muscle could be elicited. By cutting off the greater petrosal nerve at the middle cranial fossa, M-waves from the levator muscle completely disappeared on stimulating the facial nerve at the petrosal area. Results indicated that the course of the facial nerve for the levator veli palatini muscle is through the greater petrosal nerve.


Haemolytic complement in peripheral lymph of normal men. The haemolytic activity of nine individual components of complement and the concentrations of C1q, C1s, C4, C3, C3PA and C9 proteins were measured in the leg lymph and serum of four normal men. The mean lymph/serum ratio for total haemolytic complement was 0.257, for C1H50 it was 0.138, for C4H50 0.105, for C2H50 0.279, for C3H50 0.063, for C5H50 0.266, for C6H50 0.145, for C7H50 0.25, for C8H50 0.244 and for C9H50 0.253. The mean lymph/serum ratio for complement proteins was: for C1q 0.048, for C1s 0.06, for C4 0.199, for C3 0.225, for C9 0.244 and for C3PA 0.263. The low haemolytic activity of total complement and of all components in lymph seems to be dependent on the low complement protein concentration. The remarkably low lymph C1q protein concentration may play a physiological role in controlling total complement activity in lymph and interstitial fluid. The total complement haemolytic activity in lymph exceeded the haemolytic activity of C1 and C3, which may indicate the possibility of independent activation of C5-7 and C8-9. The considerably low C3H50 level, with high C3 protein concentration, at present lacks a proper explanation.


Association between susceptibility to experimental allergic encephalomyelitis and the major histocompatibility system in congenic rat strains. The genetics of the inducibility of experimental allergic encephalomyelitis (EAE) was studied in ten inbred rat strains by injection of guinea pig spinal cord in complete Freund's adjuvant. LEW rats were highly susceptible and AS2, AVN, BDV and BN rats were poorly susceptible. Congenic strains which carried the major histocompatibility haplotype of the poorly susceptible strains on the genetic background of the susceptible LEW strain were only moderately susceptible. It is concluded that major histocompatibility-linked genes exert a strong influence on the susceptibility to EAE, but that genetic background genes are also involved.


Rosette formation of human null lymphocytes with Rhesus monkey erythrocytes. Human null lymphocytes which lack B- and T-lymphocyte markers were found to form spontaneous rosettes with uncoated Rhesus monkey erythrocytes. Rosette-forming null lymphocytes were determined in null lymphocyte-enriched preparations of healthy persons and of patients with lymphoproliferative diseases. Quantitative determinations of these rosette-forming lymphocytes and other cells bearing B- or T-cell markers showed that some, and not all, of the null lymphocytes possess this rosette-forming capacity. The possibility of these null lymphocytes being related to T lymphocytes, but at a different cellular stage, is discussed.


Dynamic ventriculography in patients with ischaemic heart disease. I. Influence of nitroglycerin on left ventricular function. In 25 patients with ischaemic heart disease the effect of nitroglycerin [NTG] on segmental and global function of the left ventricle was assessed by means of left ventricular cineangiography. In 14 patients a segmental disturbance of contraction at rest was found; in six of them the function inproved after NTG. After NTG, the left ventricular diastolic volume, end-diastolic pressure, and systolic blood pressure decreased, the heart rate increased. The global left ventricular function did not significantly change. In patients with ischaemic heart disease NTG helps to differentiate reversible and irreversible disturbances of left ventricular contraction.


The cellular control of growth in cultures of Tetrahymena 2 In fluence of Fe medium and aeration on culture growth and cellular exchange of Ca. Cultures of Tetrahymena pyriformis were grown exponentially with and without the addition of Fe and with no aeration. During the prestationary growth phase of all the cultures, there was a decrease in the cellular Fe concentration in the water insoluble cell fraction (IS) containing membranes and mitochondria, simultaneous with an increase in the Ca concentration in the water soluble cell fraction (S) containing ribosomes. This has been correlated to an energy deficit in the cells at the transition to the prestationary growth phase. In spite of the ability of Fe-deficient cultures to concentrate Fe, cultures grown in media with low Fe levels soon showed the lowest cellular Fe content. The high Fe levels seen in cultures grown with no aeration may reflect cellular adaptation to a different gaseous tension in the medium. Determinations with 45Ca showed an initial, large and rapid increase in cell radioactivity which was not correlated to cellular metabolism. After this there was differentiated increase due to the metabolic status of the cells. The following sequence was seen in all the cultures: (1) an increase in the exchange of S-and (mostly) IS-Ca at the end of the exponential growth phase, (2) an accumulation of Ca in the S fraction without an increased exchange (the IS-Ca is less exchangeable), and (3) a renewed increase in the exchange of Ca when the concentration is further increased at the end of the prestationary growth phase.


Organization of the 5S RNA genes in macro- and micronuclei of Tetrahymena pyriformis. The organization of the 5S genes in macro- and micronuclei of Tetrahymena pyriformis was studied using restriction endonucleases. After complete digestion of macronuclear DNA with BamH-I or Hpa I, 5S RNA hybridized to a DNA fragment of approximately 280 base pairs (bp). When macronuclear DNA was only partially digested with these enzymes, hybridization with 32P-5S RNA demonstrated an oligomeric series with a spacing of 280 bp. These results indicate that the 5S genes are tandemly repeated in macronuclei and that the repeating unit is 280 bp (or 180,000 daltons). Since 5S RNA is 120 nucleotides, we conclude that the 5S repeat units contain a 120 bp transcribed region and a 160 bp spacer region. When macronuclear DNA was digested with Eco RI, Bgl I, or Eco RI + Bgl I, 5S RNA hybridized to DNA of molecular weight 3--4 X 10(6), suggesting that these enzymes do not cleave within a 5S repeat. These 3--4 X 10(6) dalton fragments define the maximum size of an average cluster of 5S repeated units. Assuming the size of the 5S repeat to be 0.18 X 10(6) daltons, there are about 15--20 5S repeats per average tanden cluster, and since there are 350 5S-genes per haploid genome, there must be approximately 15--20 tandem arrays. Results obtained using micronuclear DNA suggest that organization of the 5S-genes is very similar in macro- and micronuclei. Macronuclear rRNA genes are extrachromosomal palindromic dimers. In contrast, 5S genes in Tetrahymena were found to be integrated within the genomes of both macro- and micronuclei and not linked to the rRNA genes. Moreover, it is unlikely that they are palindromes; rather they appear to be tandemly repeated in "head-to-tail" linkages. Thus the organization of the 5S genes in Tetrahymena is similar to that of higher eukaryotes.


The distribution of satellite and main-band DNA components in the melanogaster species subgroup of Drosophila. I. Fractionation of DNA in actinomycin D and distamycin A density gradients. Fractionation of total adult DNA of five of the seven species of the melanogaster species sub-group of Drosophila in actinomycin D and distamycin A caesium density gradients has revealed the presence of three main-band DNA components, common to all species, and ten satellite DNAs that are distributed between the species. Satellite DNAs are either unique to a species or common to two or more species. The abundance of a common satellite DNA varies between species. There is no simple relationship between the presence of a satellite DNA and a branch point of phylogenetic divergence; nevertheless the arrangement of the species in a phylogeny that is based on the numbers of satellites held in common accurately reflects the pattern of relationships between the same species based on differences in inversions of polytene chromosomes. The species can be similarly arranged according to the compositions of their mitochondrial DNAs. It is possible that the same basic set of sequences, each of low frequency, is common to all species with arbitrary or selected amplification of particular sequences to differing extents in individual species. The conservation of satellites in the group and the close parallel between the distributions of satellites and inversions between the species suggests that either the processes that operate to change both chromosomal phenomena are similarly time-dependent and occurring at relatively low rates or that their rates of change are restricted according to some undetermined functions of these aspects of the genome.


Karyotypic evolution in an originally XY cell line of Drosophila melanogaster: a case of heterochromatin increase in vitro. The cell line Ca of Drosophila melanogaster, characterized initially by a nearly diploid and normal male karyotype (XY), was used to study chromosomal variation over a period of 5 years of cultivation in vitro. Some general aspects of cell population dynamics which are in accordance with previous findings are pointed out. Various phenomena regarding chromosomal changes leading to karyotype polymorphism are outlined, with a particular emphasis being given to the sex chromosomes. Accordingly, with the aid of fluorescence analysis, some features of the Y and the X chromosomes providing evidence of an enlargement of the heterochromatin (due to addition and to saltatory replication) are described. Moreover, a case of variation in cell morphology accompanied by karyotypic changes was observed, as well as the emergence of a new cell subline of XX type derived from the original of XY type.


The effects of naloxone on behavior maintained by cocaine and heroin injections in the rhesus monkey. The effects of repeated administration of naloxone on heroin and cocaine self-administration in non-dependent rhesus monkeys were investigated. Animals lever pressing for intravenous heroin (6 micron/kg) and cocaine (100 or 200 micron/kg) were treated for 7--10 days with naloxone at a fixed dose prior to each session. Low pretreatment doses of naloxone increased rate of responding maintained by herion. The pattern of responding over the 10-day period of treatment with the higher doses of naloxone was similar to that observed when saline was substituted for heroin. Naloxone was without effect on responding maintained by injections of cocaine.


Cytological changes of the pituitary basophils in rats slowly infused with thyrotropin-releasing hormone (TRH). Young male rats were slowly infused with synthetic TRH, 1 microgram/hr, for 1, 3, 24, 48 and 72 hr, respectively. In the control rats, the basophils of the pituitaries can be divided, in their cytological properties, into the II- (classical thyrotrophs), II/III-,III, (classical LH-cell), and III/IV-type cell. The typical IV-type cells (classical FSH-cell), however, are scarcely found in the young rats. Following 1-hr infusion of TRH, the II-type cells decrease in number with the advancement of granular release, but morphological changes are not yet concrete on the other types of basophils. The II-type cells are quickly invisible following a 3-hr infusion, while the III- and III/IV-type cells remain without any significant changes. The III- and III/IV-type cells are progressively degranulated after a 24-hr infusion. The diameter of secretory granules is reduced to 100--150 nm. The smallest ones below 50 nm in diameter, are disintegrated to disperse into the ground matrix. After degranutlaion, the III/IV-type cells appear to revert to the polygonal or stellate cells with the identical fine structure with the II-type cells. There is evidence that the thyroidectomy cells may develop from the III/IV-type cells only after a 48-hr infusion. After 72 hr, most basophils are provided with the uniform structure of "reversionary II-type cells". In reference to the high serum TSH concentration and no significant change of pituitary TSH concentration under the same experimental condition (Soji, 1978), the present author conclusively postulates that the degranulation of the III/IV-type cells may mainly reflect the conspicuous elevation of serum TSH concentration. The above morphological results are contradictory a plausible view that TRH acts only upon the thyrotrophs to release TSH. The fact that all the basophils ultimately take an appearance of "reversionary II-type cells" in the gland by the prolonged infusion of TRH may not only suggest the share of responsiveness of all the basophils to TRH, but also support the hypothesis of secretory cycle of the basophils.


Cytological changes of the pituitary basophils in rats slowly infused with LRH and with LRH and THR in combination. Young male rats were iv infused with synthetic LRH (L) and with L and TRH (T) in combination for 1, 3, 24, 48 and 72 hrs at each dose of 1 microgram/hr. All the basophils of the controls infused with saline are divided into the continuous cyclic types, i.e., II-, II/III, III-, III/IV- and III/IV/II-types. The II-, III- and III/IV-type cells correspond, in fine structure, with those of the classical thyrotrophs, LH- and FSH-gonadotrophs, respectively. A 1-hr infusion of L does not induce any serious changes in all the basophils. After a 3-hr infusion of L, the II-type cells disperse from the gland, while the III/IV-type cells diminished the number of their small secretory granules, and the lumina of endoplasmic reticulum are closed. After a 24-hr infusion, the irregularly shaped III/IV/II-type cells which may revert from the III/IV- to the II-type cells are accumulated, whose secretory granules are remarkably reduced in diameter (50--100 nm). There is evidence to show the diffusion mechanism of the secretory granules into the ground matrix. After a 48-hr infusion of L, most basophils take an appearance of III/IV/II-type cells; after a 72-hr infusion, all the basophils show the homologous fine structure. Thus, the morphological changes of the basophils following the L infusion resemble intrinsically those following the T infusion (Soji, 1976b). From these results, it is tentatively concluded that L does not act only on the III-type cells analogous to the "LH-cells", but universally upon a series of basophils. The transformations of the III/IV-type cells into the III/IV/II-type ones and those of the II-type ones into the III- or III/IV-type ones due to a slow infusion of L are inhibited to some extent by the infusion of T + L. The granular releases from the III/IV- and II-type cells due to a slow infusion of L are also inhibited to some extent by the infusion of T + L. For this reason, both T and L may act, not synergistically but antogonistically, upon the transformation of a series of basophils and their granular release.


The causative organism of contagious equine metritis 1977: proposal for a new species to be known as Haemophilus equigenitalis. The aetiological agent of contagious equine metritis (CEM) has been investigated bacteriologically in a wide range of cultural and conventional biochemical tests, in the eletron microscope, for DNA base composition (36.1 per cent GC), for susceptibility to various antimicrobial agents and antigenically by means of tube and slide agglutination tests. The organism is a fastidious, Gramnegative, non acid-fast coccobacillus which in biochemical tests is very unreactive. In conventional tests, only the oxidase, catalase and phosphatase tests were positive. Dependance on neither X nor V factors could be demonstrated, but some stimulation of growth by X factor was observed. The organism could not be identified with any known species and even allocation to an appropriate characters, we propose the organism as a new species of the genus Haemophilus: H. equigenitalis, type strain NCTC 11184 (61717/77).


Calcium uptake in skeletal muscle mitochondria. I. The effects of chelating agents on the mitochondria from fatigued rats. Female Wistar rats were used to determine the effects of the chelating agents, EDTA and EGTA, on the in vitro 45Ca2+ accumulation by mitochondria isolated from the skeletal muscle of fatigued animals. The rats were divided into three groups: sedentary-rested (SR), trained-rested (TR), trained-exhausted (TE). The trained groups were exercised on a treadmill for 1 h daily, five times a week, for 22 weeks. At the conclusion of the training program, the TE group was rapidly exercised to exhaustion immediately following their daily 1-h run. In the TR group EDTA reduced 45Ca2+ binding while both EDTA and EGTA appeared to increase mitochondrial Ca2+ and Mg2+ content. In the TE group, EDTA reduced endogenous mitochondrial Ca2+ and Mg2+ content, while both EDTA and EGTA increased 45Ca2+ binding. Since chelating Ca2+ and Mg2+ from the membrane may affect the structure and function of the mitochondria, it is suggested that the use of chelating agents during the isolation of mitochondria from the skeletal muscle of trained rats be viewed with caution.


Three forms of DNA polymerase from Drosophila melanogaster embryos. Purification and properties. DNA polymerase was purified from Drosophila melanogaster embryos by a combination of phosphocellulose adsorption, Sepharose 6B gel filtration, and DEAE-cellulose chromatography. Three enzyme forms, designated enzymes I, II, and III, were separated by differential elution from DEAE-cellulose and were further purified by glycerol gradient centrifugation. Purification was monitored with two synthetic primer-templates, poly(dA) . (dT)-16 and poly(rA) . (dT)-16. At the final step of purification, enzymes I, II, and III were purified approximately 1700-fold, 2000-fold and 1000-fold, respectively, on the basis of their activities with poly(dA) . (dT)-16. The DNA polymerase eluted heterogeneously as anomalously high-molecular-weight molecules from Sepharose 6B gel filtration columns. On DEAE-cellulose chromatography enzymes I and II eluted as distinct peaks and enzyme III eluted heterogeneously. On glycerol velocity gradients enzyme I sedimented at 5.5-7.3 S, enzyme II sedimented at 7.3-8.3 S, and enzyme III sedimented at 7.3-9.0 S. All enzymes were active with both synthetic primer-templates, except the 9.0 S component of enzyme III, which was inactive with poly(rA) . (dT)-16. Non-denaturing polyacrylamide gel electrophoresis did not separate poly(dA) . (dT)-16 activity from poly(rA) . (dT)-16 activity. The DNA polymerase preferred poly(dA) . (dT)-16 (with Mg2+) as a primer-template, although it was also active with poly(rA) . (dT)-16 (with Mn2+), and it preferred activated calf thymus DNA to native or heat-denatured calf thymus DNA. All three primer-template activities were inhibited by N-ethylmaleimide. Enzyme activity with activated DNA and poly(dA) . (dT)-16 was inhibited by K+ and activity with poly(rA) . (dT)-16 was stimulated by K+ and by spermidine. The optimum pH for enzyme activity with the synthetic primer-templates was 8.5. The DNA polymerases did not exhibit deoxyribonuclease or ATPase activities. The results of this study suggest that the forms of DNA polymerase from Drosophila embryos have physical properties similar to those of DNA polymerase-alpha and enzymatic properties similar to those of all three vertebrate DNA polymerases.