K+ conduction description from the low frequency impedance and admittance of squid axon. The form of power spectra of K+ conduction fluctuations in patches of squid axon suggested that K+ conduction kinetics are higher than first order (Fishman, Moore & Poussart,  1975, J. Membrane Biol. 24:305). To obtain an alternative description of ion conduction kinetics consistent with spontaneous fluctuations, the complex impedance and admittance of squid (Loligo pealei) axon were measured at low frequencies (1-1000 Hz) with a four electrode system using white Gaussian noise as a stochastic perturbation. As predicted from the spontaneous noise measurements, a low frequency impedance feature is observed between 1 and 30 Hz which is voltage and temperature dependent, disappears after substantial reduction in [Ki+], and is unaffected by the state of Na+ conduction or active transport. These measurements confirm and constitute strong support for the patch noise measurements and interpretations. The linearized Hodgkin-Huxley (HH) equations do not produce the low frequency feature since first order ion conduction kinetics are assumed. Computation of diffusion polarization effects associated with the axon sheath gives a qualitative account of the low frequency feature, but the potential dependence is opposite to that of the data. Thus, K+ conduction kinetics in the axon are not adequately described by a single first order process. In addition, significant changes in HH parameter values were required to describe the usual impedance (resonance) feature in Loligo pealei axon data.


Passive sodium fluxes across toad bladder in the presence of simultaneous transepithellal gradients of concentration and potential. Bidirectional sodium fluxes were measured across toad bladder sacs after eliminating active transport with ouabain. Transepithelial potential was clamped to 100 mV or the Nernst potential, psieq, at varying sodium concentrations, Cm, in the mucosal medium. Serosal sodium concentration, Cs, was held constant. Equations were derived for permeability, partial ionic conductance, and unidirectional fluxes as functions of Cm and Cs, based in part on the assumption that the ratio, Q, of bulk sodium permeability to tracer sodium permeability is a constant, independent of concentration and potential. The results conformed closely to these equations.


In vitro studies of skeletal muscle membranes. Adenylate cyclase of fast and slow twitch muscle and the effects of denervation. Sarcolemmal membranes were prepared from slow-twitch (red) and fast-twitch (white) skeletal muscle of the rat. A sensitive adenylate cyclase assay was used and basal, fluoride- and catecholamine-stimulated activities measured. The greater in vivo sensitivity of red muscle to the effects of catecholamines correlates, in the present study, with approximately a twofold stimulation of its sarcolemmal adenylate cyclase with isoproterenol (10 micronm). The white muscle enzyme, on the other hand, is only minimally stimulated (20%) at the same concentration of beta-adrenergic agonist. Fast-twitch muscle is known to be physiologically insensitive to catecholamine in vivo. A course of sciatic nerve denervation was followed to further distinguish these two metabolic types of skeletal muscle and their respective adenylate cyclases. The slow-twitch muscle enzyme activities were completely and permanently lost on denervation. The white muscle enzyme, however, recovered almost completely after an initial reduction in specific activity the first week. Interestingly, the NaF-stimulated activity lagged behind both the basal and hormone-stimulated activities of the white muscle enzyme, in returning to control levels. The activities of cyclic nucleotide phosphodiesterase were evaluated in homogenates of the two muscle types in innervated rats and following denervation, in order to further define the neural influence on skeletal muscle cyclic nucleotide metabolism. The results suggest that the motor nerve may regulate some of the metabolic properties of slow-twitch muscle (which may involve cyclic AMP) by controlling the responsiveness of its sarcolemmal-bound adenylate cyclase system.


A gradient of diffusible substance in a monolayer of cultured cells. Monolayers of hypoxanthine phosphoribosyl transferase-deficient and their corresponding wildtype cells have been placed adjacent to each other with a newly described method. Autoradiographs from such preparation after incubation with 3H-hypoxanthine allow the direct visualization of gradients of incorporated radioactivity at the border between the two cell types. The gradients can be described by an exponential function, and the amount of radioactivity incorporated decreases to less than 1% at a distance of 1 mm from the wild-type cells. A possible mechanism to convert exponential gradients to linear ones over a certain concentration range is discussed.


Changes in absorption, fluorescence, dichroism, and Birefringence in stained giant axons: : optical measurement of membrane potential. The absorption, fluorescence, dichroism, and birefringence of stained squid axons were measured during action potentials and voltage clamp steps in an effort to find large optical signals that could be used to monitor membrane potential. Changes in all four optical properties were found that were linearly related to membrane potential and, with several new dyes, the signal-to-noise ratios were larger than any obtained previously. The problem of photodynamic damage was greatly diminished; with a merocyaninerhodanine dye, the photodynamic damage associated with intense light and the presence of oxygen was negligible. The absorption change obtained with this dye was relatively large; it could be measured with a signal-to-noise ratio of 100:1 during a single action potential.


Effect of turgor pressure on water permeability of Allium cepa epidermis cell membranes. Using onion epidermis layer a very accurate method for measuring the permeability of epidermis cells to water was standardized. In this method a 1.4 cm diameter epidermis disc was soaked in tritiated water (50 micronCi/ml) for about 1 hr. Next the disc was mounted in a specially designed elution chamber where it was held flat and washed on the noncuticular side with ordinary water. A constant flow rate, high enough to minimize unstirred layer effect, was used. Permeability was calculated in the usual way after separating different exponentials from the efflux curve of tritiated water. Turgor pressure of the cell was regulated by soaking thedisc in mannitol solutions containing tritiated water and washing it in the chamber with same concentration mannitol solution containing no radioactivity. Water permeability values were found to decrease less than 8% when the turgor pressure was decreased from 8 atm (full turgor) to zero. Turgor pressure had no significant effect on the water permeability of onion epidermal cells. Our results are contradictory to the findings of Zimmerman and Steudle (1974, J Membrane Biol. 16:331) but aresimilar to the findings of Tazawa and Kamiya (1966, Aust J. Biol. Sci. 19:399) and Kiyosawa and Tazawa (1972, Protoplasma 74:257).


Role of plasma membrane phospholipids in the uptake and release of transferrin and its iron by reticulocytes. The involvement of membrane phospholipids in the utilization of transferrin-bound iron by reticulocytes was investigated using [59Fe]- and [125I]-labelled transferrin and rabbit reticulocytes which had been incubated with phospholipase A. Transferrin and iron uptake and release were all inhibited by phospholipase A which produced a marked decrease in the relative abundance of phosphatidylcholine and phosphatidylethanolamine and equivalent increases in their lyso-compounds in the reticulocyte plasma membrane. There was a close correlation between the iron uptake rate and the rate and amount of transferrin uptake and the amount of the lysophospholipids in the membrane. Incubation of the cells with exogenous lysophosphatidylethanolamine or lysophosphatidylcholine also produced inhibition of iron and transferrin uptake. The reduced uptake produced by phospholipase A could be reversed if the lyso-compounds were removed by fatty acid-free bovine serum albumin or by reincubation in medium 199. Treatment with phospholipase A was shown to increase the amount of transferrin bound by specific receptors on the reticulocyte membrane but to inhibit the entry of transferrin into the cells. The present investigation provides evidence that the phospholipid composition of the cell membrane influences the interaction of transferrin with its receptors, the processes of endocytosis and exocytosis whereby transferrin enters and leaves the cells, and the mechanism by which iron is mobilized between its binding to transferrin and incorporation into heme. In addition, the results indicate that phosphatidylethanolamine is present in the outer half of the lipid bilayer of reticulocyte membrane.


Cold-induced hemolysis in a hypertonic milieu. Suspension of human erythrocytes at 37 degrees C in an environment made hypertonic by increasing concentrations of sodium chloride and sucrose was followed by hemolysis when the temperature was lowered to 0 degrees C. Two distinct stages were involved in this hemolytic phenomenon, the first being incubation with hypertonic solute at some temperature above 20 degrees C with an increasing effect up to 45 degrees C, and the second stage consisting of lowering the temperature below 15 degrees C with increasing hemolysis down to 0 degrees C. The rate of cooling was not an important factor, but the presence of ions reduced the extent of cold-induced hemolysis in hypertonic sucrose. No significant release of membrane phospholipid and cholesterol accompanied this hemolysis. The solubilization of membrane protein components was investigated, with some differences appearing on sodium dodecyl sulfate polyacrylamide gel electrophoresis between hypertonic and isotonic supernatants. Spectrin could not be identified in solubilized form. Correlation of the temperatures of note in these studies with results from the literature on other biological effects of temperature-induced phase transitions in membrane lipids strongly points to the conclusion that such transitions are involved in the mechanism of cold-induced hypertonic hemolysis. It is postulated that the hypertonic milieu has resulted in membrane-protein alteration damage which prevents normal adaption to the new physical state of the membrane lipids during cooling.


Study of amino and sulfhydryl sites in the sodium pathway in dog red blood cell membranes. Amino reactive TNBS (2,4,6-trinitrobenzene sulfonic acid). SITS (4-acetamido-4'-isothiocyano-stilbene-2-2' -disulfonic acid), and Zn++, and SH reactive Hg++ were employed to study sodium channels in dog red blood cells. Simultaneous modification of the membrane with both a SH and an amino modifier results in an increase in Na+ permeability which is equal to the sum of their individual effects. This indicates that SH and amino sites are separate units. Three lines of evidence indicate that the amino sites are more superficial than the SH sites. (1) Pretreatment with an amino modifer decreases the effectiveness of subsequent SH modification. (2) SITS, a nonpenetrating amino reagent, enhances Na+ permeability while DTNB, a nonpenetrating SH modifier, is ineffective. (3) Pretreatment of amino sites decreases the apparent affinity of Hg++ for SH sites. In addition, three lines of evidence indicate that TNBS and Zn++ modify different amino sites. First, simultaneous modification with TNBS and Zn++ results in an increase in Na+ permeability equal to the sum of their individual effects. Secondly, Zn++ causes an increase in Na+ permeability in cells previously treated with TNBS. Finally, the pH dependence of Zn++ modification is oposite that for TNBS modification. These pH experiments suggest than Zn++ enhances Na+ permeability by reacting with unprotonated amino sites while TNBS modifies protonated amino sites. It is concluded that the sodium permeability of dog red blood cells is normally limited by superficial amino sites and deeper slfhydryl sites in the sodium channels.


Intracellular gradients of electrical potential in the epithelial cells of the Necturus gallbladder. When single-barrelled electrodes (5-60 Momega) were advanced under manual control from the mucosal side of the epithelium the mucosal membrane was on average indented by about 40 micron before the microelectrode pentrated the cell. Since this dimpling was comparable with the total depth of the cell, which recovered its original shape within 0.5 sec, the steady intracellular potential was recorded only about 14 micronm from the basal (serosal) membrane. Fast recording of the associated change in potential revealed an abrupt drop to -26 mV at a mean rate of 84 V/sec, followed by a further slow drop to a steady value of about -50 mV at a mean rate of 0.28 V/sec. The initial level of -26 mV may be regarded as the potential difference across the mucosal membrane. This conclusion was confirmed by mounting the microelectrode on a piezoelectric probe, which delivered 3 micron jabs in less than 0.5 msec. With this device in operation to prevent dimpling, the mean potential difference across the mucosal membrane was recorded as -29 mV. In all cases the potential across the basal membrane was recorded as -52 mV. Manual advance of the microelectrode tip within the cytoplasm yielded an intracellular potential gradient of 0.6 mV/micron. The same potential profile and membrane potentials were demonstrated on penetrating the epithelium from the serosal side, and measurements with multibarrelled electrodes whose tips were staggered in depth gave roughly the same internal potential gradient. The resistivity of the cytoplasm was determined by a triple-barrelled microelectrode, and varied from 10 times that of Necturus saline at the mucosal end of the cell to 4 times in the middle and 6 times at the serosal end.


Synthesis of tritiated 4,4'-diisothiocyano-2,2'-stilbene disulfonic acid ([3H]DIDS) and its covalent reaction with sites related to anion transport in human red blood cells. The potent and specific inhibitor of anion permeability, 4,4'-diisothicyanostilbene-2,2'-disulfonic acid (DIDS) was synthesized in tritiated form ([3H]DIDS) from tritiated 5-nitrotoluene-o-sulfonic acid. Its reactions with and effects on red blood cells were compared with those of a reduced form ([3H]H2DIDS), previously used as a tracer for DIDS. The rate of covalent reaction of [3H]DIDS was substantially faster than that of [3H]H2DIDS at all temperatures tested. With both agents, the rate of reaction was increased in alkaline media, although the response occurred at a lower pH with [3H]DIDS. On the other hand, the relationship of irreversible membrane binding to the degree of inhibition of sulfate fluxes was linear and virtually the same for both agents, with 100% inhibition associated with the binding of approximately 1.2 X 10(6) molecules per cell. About 90% of the binding for each probe was to a particular membrane protein, known as band 3, equivalent to about 1 mole of agent per mole of protein.


Active sodium transport and the electrophysiology of rabbit colon. The electrophysiologic properties of rabbit colonic epithelial cells were investigated employing microelectrode techniques. Under open-circuit conditions, the transepithelial electrical potential difference (PD) averaged 20 mV, serosa positive, and the intracellular electrical potential (psimc) averaged -32 mV, cell interior negative with respect to the mucosal solution; under short-circuit conditions, psimc averaged -46 mV. The addition of amiloride to the mucosal solution abolishes the transepithelial PD and active Na transport, and psimc is hyperpolarized to an average value of -53 mV. These results indicate that Na entry into the mucosal cell is a conductive process which, normally, depolarized psimc. The data obtained were interpreted using a double-membrane equivalent electrical circuit model of the "active Na transport pathway" involving two voltage-independent electromotive forces (emf's) and two voltage-independent resistances arrayed in series. Our observations are consistent with the notions that: (a) The emf's and resistances across the mucosal and baso-lateral membranes are determined predominantly by the emf (64 mV) and resistance of the Na entry process and the emf (53 mV) and resistance of the process responsible for active Na extrusion across the baso-lateral membranes: that is, the electrophysiological properties of the cell appear to be determined solely by the properties and processes responsible for transcellular active Na transport. The emf of the Na entry process is consistent with the notion that the Na activity in the intracellular transport pool is approximately one-tenth that in the mucosal solution or about 14 mM. (b) In the presence of amiloride, the transcellular conductance is essentially abolished and the total tissue conductance is the result of ionic diffusion through paracellular pathways. (c) The negative intracellular potential (with respect to the mucosal solution) is due primarily to the presence of a low resistance paracellular "shunt" pathway which permits electrical coupling between the emf at the baso-lateral membrane and the potential difference across the mucosal membrane; in the absence of this shunt, the "well-type" electrical potential profile characteristic of rabbit colonic cells would be 'converted' into a "staircase-type" profile similar to those reported for frog skin and toad urinary bladder by some investigators.


Pulsed nuclear magnetic resonance study of 39K within halobacteria. The 39K contents of isolated pellets and supernatant solutions from suspensions of Halobacterium halobium were studied at 21-22 degrees C by pulsed NMR spectroscopy. The rates of transverse relaxation were measured directly from the free induction decay (FID). The rate of longitudinal relaxation was measured by studying the FID after pairs of pulses of approximately 90 degrees. Care was exercised to minimize the effect of magnetic field inhomogeneity; its contribution to the FID was approximately 25-30 sec-1. The transverse relaxation process was found to consist of at least two components, whose rates were 321-449 sec-1 and 1,122-2,067 sec-1. In one preparation where the longitudinal relaxation process was studied, the data could be well fit to a single exponential relaxing at 253 +/- 33 (mean +/-95% confidence limits) sec-1. Comparison of the relative intensities of the NMR signals with the results of atomic absorption photometric analyses indicated that the great bulk of the intracellular 39K was detected by the NMR techniques used. The data obtained from the current NMR of H. halobium are consistent with: (1) fractional binding of less than 3% of the total intracellular K+, (2) a small ordering factor characterizing all of the intracellular K+, or (3) some combination of the two.


Chromatin diminution in early embryogenesis of Ascaris lumbricoides L. var. suum. The occurrence of chromatin diminution in early Ascaris lumbricoides L. embryos has been studied in detail, and it is shown that it is possible to preselect three characteristic types of mitoses: pre-diminution, diminution, and post-diminution mitosis. The first three embryonic mitotic divisions are of the pre-diminution type. Chromatin diminution occurs after the third mitosis, but there is a variation from embryo to embryo as to whether or not chromosomal diminution occurs during the fourth, fifth, and six divisions. However, the seventh embryonic division, which gives rise to an eight-cell embryo, always exhibits chromatin diminution. Subsequent mitoses of somatic cells already in the diminished state are of the post-diminution type of mitosis.


A new ganglion in male Moniliformis moniliformis (Acanthocephala). A new ganglion identified as the bursal ganglion is described from male Moniliformis moniliformis. This ganglion is located adjacent to the pseudocoel and longitudinal muscle fibers and medial to the dorsal lacunar canal about 1 mm from the posterior end of males with non-everted bursa. The ganglion consists of four large club-shaped cells with single nuclei and bipolar neurons. The ganglion cells are paired with one neuron from each cell innervating the opposite side of the worm.


Embryogenesis of Xyleborus ferrugineus (Fabr.) (Coleoptera, Scolytidae). I. External morphogenesis of male and female embryos. A description of the external morphogenesis of male and female embryos of X. ferrugineus, derived from in vivo observation, is presented here. The embryogenesis of this beetle is typical of the Coleoptera, and is also similar in most respects to the generalized insect plan. Observed unusual aspects of X. ferrugineus embryogenesis include the precocious formation of primordial g/rm cells and their temporary exclusion from somatic blastoderm; the precocious in situ delineation of gnathal metameres, the closing of the gastral groove beginning at both ends and proceeding towards its middle, the formation of several paired lateral amnio-serosal folds, the formation of cellular processes between the amnion and serosa, and the absorption of the thoracic limbs to produce an apodous larva. The embryonic developmental picture provided here, and the previously established means of rearing X. ferrugineus in controlled laboratory culture jointly provide a promising basis for the further use of this insect in developmental studies.


Embryogenesis of Xyleborus ferrugineus (Fabr.). (Coleoptera, Scolytidae) II. Developmental rates of male and female embryos. The developmental rates of male and female embryos of Xyleborus ferrugineus were compared by charting for each sex the mean age for each of ten discrete morphological stages of embryogenesis from pole cell exclusion to eclosion male and female embryos developed synchronously from stage 1 (which begins with pole cell exclusion) through stage 4 (which ends with the completion of germ band extension and metamerization). After stage 4 and throughout the remainder of embryogenesis, the mean ages per morphological stage of male embryos were significantly greater than those for female embryos. The expected physiological consequences of a haploid genome in the male embryo as compared to a diploid condition in the female embryo are discussed as the possible basis for the persistent lag observed in male developmental rate after stage 4.


Phylogenetic studies of two rubredoxins from sulfate reducing bacteria. The sequences of two rubredoxins isolated from the sulfate reducing bacteria: Desulfovibrio vulgaris and Desulfovibrio gigas have been elucidated. They have similar sequences but many more differences occur than would be expected from two bacteria of the same genus. Of the 52 sites, only 37 are occupied by identical residues. The primary structures are compared with those of the anaerobic bacteria rubredoxins of Clostridium pasteurianum, Micrococcus aerogenes, Pseudomonas oleovorans and Peptostreptococcus elsdenii: only 12 identities are found, mostly in the two clusters that contain two iron-bound cysteines each. A phylogenetic tree based on the primary structures is presented and possible relations with plant and bacterial ferredoxins are discussed. A secondary and tertiary structure stereochemically compatible with the sequence data, is proposed.


Alignment statistic for identifying related protein sequences. Closely related proteins show an obvious kinship by having numerous matching amino acids in their aligned sequences. Kinship between anciently separated proteins requires a statistical evaluation to rule out fortuitous similarities. A simple statistic is developed which assumes equal probability for all codon pairs, and a table of critical values for amino acid sequence alignments of lengthnments of length 200 or less is presented. Applying this statistic to V and C regions of immunoglobulin chains, aligned on the basis of shared features of three-dimensional structure, provides evidence that the V and C sequences descended from a common ancestor. Similarly the distant evolutionary relationship of dehydrogenases, flavdoxin, and subtilisin, suggested by structural alignments, is verified. On the other hand, the statistic does not verify a common evolutionary origin for the heme binding pocket in globins and cytochrome bs. Empirical evidence from the distribution of MMD values of amino acid pairs in comparisons of misaligned polypeptide chains and from Monte Carlo trials of sequences aligned with arbitrary gaps supports the validity of the statistic.


Growth of Klebsiella aerogenes on xylitol: implications for bacterial enzyme evolution. When Klebsiella aerogenes was grown in continuous culture with xylitol. an unnatural pentitol, as the growth limiting substrate, the structural gene which codes for ribitol dehydrogenase, an enzyme which gratuitously catalyzes the oxidation of xylitol to D-xylulose, was duplicated. It appears that the duplication mechansim only duplicates the gene which is subjected to selective pressure and not any of the other closely linked genes. The degree to which the ribitol dehydrogenase gene is duplicated does not appear to be strictly correlated with the ability to grow faster on xylitol. Duplication mutants do, in fact, grow faster than their parent strain, but when challenged to grow at even higher growth rates there is a catabolic repression of enzyme activity. Thus a situation is created in which a structural gene is duplicated in response to selective pressure; these mutants can grow faster on the new substrate, but faster growth results in a "silencing" of a portion of the genes by catabolite repression.


Standard error of immunological dating of evolutionary time. The empirical variance of the immunological distance as measured by microcomplement fixation with albumin is determined. The variance obtained is at least two times larger than the mean when the mean is small and the ratio of the variance to the mean increases with increasing mean. Thus, the immunological dating of evolutionary time has a large standard error. It is shown that in bird lysozymes the relationship between immunological distance (y) and the number of amino acid substitutions per 100 sites (x) is given by y = 4.2 x approximately.


Novel prebiotic systems: nucleotide oligomerization in surfactant entrapped water pools. Oligomerization of 5' -TMP in water pools entrapped by dodecyl-ammonium chloride surfactant aggregates in benzene: hexane in the presence of dicyanodiimide at temperatures ranging from 21 degree -72 degree resulted in the formation of linear and cyclic oligonucleotides containing up to pentamers. Effects of temperature, time and surfactants have been examined. Rate constants for the formation of oligomers have been determined at five different temperatures. These data afforded values of (formula: see text). Prebiotic significance of these results are discussed.


Hemoglobin and the genetic code. Evolution of protection against somatic mutation. One-half of the twenty amino acids of the genetic code are just one mutational step away from the chain-terminator codons UAA, UAG, and UGA. It is postulated that somatic mutation to terminator is a hazard to which the organism has and to respond by adjusting certain proteins in the direction of fewer mutable residues. This view is supported by calculations based on the primary structure of five of the human hemoglobin chains. Each chain is scored for mutability to terminator in accord with the numbers and kinds of amino acids present. Among the adult chains, the most essential one, the alpha, has lowest mutability. The beta and delta follow, and in order of the presumed harm to the organism of a shortage of chain copies. Ante-natal chains tend to have higher mutabilities, supporting the view that cumulative mutational change in DNA can do little if the gene ceases to transcribe early in life. Two other predicitons based on the supposition of effective selection against mutability to terminator are also met: chain length of polypeptides is negatively correlated with their scores for mutability to terminator, and examination of the recently determined sequence of beta messenger RNA shows preferential use of codons that are not readily mutable to terminator.


The amino acid sequence of dog (Canis familiaris) hemoglobin. The manual sequencing of the tryptic peptic from the alpha and beta chains of dog hemoglobin is described, including evidence for the existence of two alphaT-13 peptides and thus 2 alpha chains, one with threonine and one with alanine at position 130. Although the actual sequence was published in 1970, the evidence on which it was based has not previously appeared.


On the tryptic peptides from hemoglobin chains of six carnivores. The amino acid compositions of the tryptic peptides of the following carnivore hemoglobin chains have been determined: gray fox (Urocyon cineroargenteus); raccoon (Procyon lotor); polar bear (Thalarctos maritimus); coati mundi (Nasua nasua) beta chain; coati mundi (Nasua narica) two beta chains; cat (Felis catus) alpha chain; and lion (Pantbera leo) beta chain. These provide a basis for future sequencing of these hemoglobins and construction of an evolutionary tree. The specific results are summarized in the following article (Stenzel and Brimhall, 1977).


The tryptic peptide composition of the beta chains of hemoglobins A and B of the Domestic cat (Felis catus). The tryptic peptides from the betaA and betaB chains of cat hemoglobins A and B have isolated and the amino acid compositions determined. Differences between the two chains were found in two peptides, betaT-1 (Glylead toSer) and betaT-14 (Asnlead toSer and Lyslead toArg). The Glylead toSer and Lysleand toArg substitutions areplaced at beta-1 and beta-144 respectively from earlier work, and the third substitution, AsnleadSer at beta-139 is suggested from this work. in addition, the presence of a blocked amino terminus in betaB has been confirmed. Tentative sequences constructed by homology with known beta-chain structures suggest the occurrence of substitutions at alpha1beta1 contacts in betaA and betaB that may be functionally significant. There are at least 18 differences in amino acid composition between cat betaA and dog beta-chains and 22 differences between cat betaA and normal adult human beta-chains.


Preliminary survey of carnivore hemoglobin compositions. Conclusions. The amino acid differences among 12 beta chains and 10 alpha chains of carnivore hemoglobins are given. These hemoglobins conform to the substitution rate found for hemoglobins in general. A table compares the differences among known alpha-chain sequences when deduced from tryptic petide compositions and when taken from actual sequences. Among the carnivore hemoglobins studies, tryptic peptide compositions are about 14 % low in giving the number of sequence differences.


Nearest-neighbor doublets in protein-coding regions of MS2 RNA. 'Nearest neighbor' base pairs (doublets') in the protein-coding regions of MS2 RNA have been tabulated with respect to their positions in the first two bases of amino acid codons, in the second two bases, or paired by contact between adjoining codons. tconsiderable variation is evident between numbers of doublets in each of these three possible positions, but the totals of each of the 16 doublets in the coding regions of the MS2 RNA molecule show much less varation. Compila-acid composition of proteins coded by such strands.


Ego deficiencies in the areas of pleasure, intimacy, and cooperation: guidelines in the diagnosis and treatment of sexual dysfunctions. Psychopathology, in the form of a deficiency in the ego functions of pleasure, intimacy, or cooperation, is usually associated with sexual dysfunctions that prove refractory to sex therapy techniques alone. Each of these deficiencies forms a distinct clinical syndrome with specific treatment requirements, necessitating further diagnostic subcategorization of sexual disorders.


Sex therapy as an aid to marital and family therapy. The treatment by a qualified sex therapist of a sexually dysfunctional couple referred by a family or marital therapist often significantly factilitates the overall therapeutic progress. When "successful," relevant effects of sex therapy include a better dyadic relationship, optimism about therapy in general, and greater willingness to change, beneficial ripple effects on other family members often occur. Resistance by partners or other family members is considered, as is the use of "unsuccessful" sex therapy as a diagnostic tool. Questions as to whether sex therapy should follow, interrupt, or proceed concurrently with family/marital therapy are discussed.


Hypoactive sexual desire.

Hypoactive sexual desire. The sex therapy literature has concentrated on disorders of the excitement and orgasm phases of the sexual response. However, disorders of sexual desire have been virtually neglected, although low-libido disorders are highly prevalent, may be extremely distressful to patients and their partners, and influence the course and prognosis of therapy. This paper focuses on this important aspect of human sexuality. Some clinical features of hypoactive sexual desire are described, and some hypotheses about etiology and prognosis are presented.


Seminars for separated/divorced: an educational approach to postdivorce adjustment. This paper describes a seminar approach to postdivorce adjustment. Topical content areas covered in the 7-week seminars are discussed. The interaction of leader-guided problem solving and participant input is addressed along with a series of preliminary conclusions reached concerning the efficacy of this approach to treatment.


Resisting the pressure to become a surrogate: a case study. Masters and Johnson's discussion of their surrogate program and informal interviews with surrogates reveal that the surrogate role is stressful. The surrogate hopes to bypass explosive relationship issues in order to lower the patient's level of tension. Such bypassing is difficult, sometimes impossible, and of limited therapeutic value. An alternative tactic has been developed for Individual Sex Therapy and is exemplified by a case study. The case described involves an involuntary ejaculator who tried to sidestep the pressure that he felt from relationships in general. This case exemplifies the pressure in the Individual Sex Therapy situation, how the surrogate approach attempts unsuccessfully to bypass this pressure, and the processes by which the pressure can be countered.


On the etiology of sexual dysfunction.

On the etiology of sexual dysfunction. Lack of consideration of the sexually functional population has led to misconceptions about the causes of both sexual dysfunction and sexual functioning. Although observation of the dysfunctional population appears to indicate that performance anxiety is the first-order cause of dysfunction, it is proposed that in the functional population performance anxiety has the opposite effect, generating automatic functioning rather than dysfunction. The first-order cause of dysfunction is then whatever causes this differential response to performance anxiety, conceivably an inability or unwillingness to bypass pathogenic influences. Thus automatic functioning can mask the effects of all pathogenic influences on sexuality, making these effects appear random, confounding etiological issues and creating the belief that the causes of the sexual dysfunction and sexual disorder cannot be specified.


Some properties of cells cultured from early-lactation human milk. Cells that can be cultured from pools of early-lactation milk were studied. Under the culture conditions used, the majority of cells attached to collagen-coated dishes; most of these remained single, did not divide, and in their adhesiveness, phagocytic ability, and ultrastructure resembled macrophages or histiocytes. On a plate seeded with approximately 3 X 10(5) cells, however, 10-100 colonies of dividing cells developed. These cells had the junctional complexes typical of epithelial cells and grew well in a medium supplemented with human serum and hydrocortisone for 16-20 days after seeding. After removal of serum from the medium, some cells continued to traverse the cell cycle, and the colonies containing these cells were morphologically distinct from those which became quiescent. The nondividing cells in milk could be separated from the milk epithelial cells and were able to stimulate the growth of epithelial cultures from benign mammary dysplasias.


Spontaneous thymoma in an inbred strain of rat. Thymomas that appear with high frequency in inbred WAB rats living into old age were examined. The spontaneous thymomas described were discrete, encapsulated bodies composed of lymphocytes and epithelial cells, with a predominance of lymphocytes. The rat neoplasms resembled the human thymomas, and their rate of incidence in older animals may suggest their use as models for the human condition.


Chromosome aberrations and sister chromatid exchanges in Chinese hamster cells exposed to various chemicals. To evaluate its usefulness for detection of mutagenicity and oncogenicity of chemicals, the frequency of chromosome aberrations and sister chromatid exchanges (SCE) was examined in cultured Chinese hamster cells exposed to 33 kinds of chemical agents, including 2 aromatic hydrocarbons, 7 amine derivatives, 3 phenols, 6 carboxylic acid derivatives, 4 azo compounds, 7 heterocyclic compounds, and 4 other types of agents. Among them, 7 chemicals, including 5 known oncogens (N-n-butyl-N-nitrosourethan, N-methyl-N-nitrosourea, beta-propiolacton, methylazoxymethanol acetate, and propane sultone) and 2 non-oncogens (potassium sorbate and sodium benzoate) were inducers of aberrations and/or SCE, although the effective doses for the last two agents were considerably higher. The mutagenic activities estimated by chromosome aberrations were not necessarily the same for a given chemical when estimated by SCE. Slightly to moderately increased rates of SCE or aberrations were noticed among the other 26 chemicals. However, these values were usually without a clear dose relationship.


Properties of RBL-5 leukemia cells cultivated in vitro. RBL-5 leukemia induced by Rauscher murine leukemia virus (R-MuLV) carried in the ascites form in C57BL/6 mice was cultivated in vitro and passaged continuously. Both low- and high-passage cells released the same level of murine leukemia virus, as detected by the XC plaque assay. Membrane antigens of the cultivated cells were also detected by the indirect immunofluorescence assay. Both low- and high-passage cells showed the same activity of antigen detectable by anti-R-MuLV serum, whereas only the high-passage cells reacted strongly against a syngeneic antiserum to RBL-5 crude membrane (CM). The oncogenicity of the low and high passages of the cultivated cells was compared with the ascites cells. The results indicated that the oncogenic potential of the cells gradually decreased during in vitro passage. The accumulation or retention of CM antigen on the membrane of the high-passage cells suggested that their decreased oncogenic potential might be due to increased immunogenicity and/or immunosensitivity. This strengthened the hypothesis that the cellular transplantation antigens are not identical to type C viral antigens, since the R-MuLV antigens were not altered during passage in vitro.


Survival curve of a human melanoma in nude mice. Using cells from our tissue culture of human melanoma cell line Na 11, we transplanted 1 X 10)6) tumor cells sc into athymic nude mice. Tumors appeared after a latent period of 4-10 days; when they reached a mean volume of 100 mm3 we irradiated them with various doses of X-rays. Some tumors were irradiated while the mice were still alive; others were treated 10 minutes after the animals had been asphyxiated with nitrogen. All irradiation was done in the presence of oxygen. These tumors were excised, and cell suspensions were prepared; the cells formed colonies with a mean plating efficiency of 29%. In another series of experiments, we irradiated tumor cells in vitro 2 hours after excision, when most cells were fixed and presumably oxygenated. We then calculated survival curves for the tumor cells irradiated under these three conditions and found an average anoxic cell fraction of 85%, which was much higher than that reported in many other tumor systems. We explored several possible explanations for this phenomenon.


R3327 adenocarcinoma of the Copenhagen rat as a model for the study of the immunologic aspects of prostate cancer. The Dunning R3327 prostate adenocarcinoma of the Copenhagen rat was developed as a suitable model of human prostate cancer. Inoculation of tumor tissue mince or cells sc in the flanks of recipient rats produced tumors that had the macroscopic and microscopic characteristics of the human disease. The histologic picture of these tumors was that of a well-differentiated adenocarcinoma with the formation of glands and acid secretions within the acini. Tumors were also produced in the dorsolateral lobe of the prostate by the injection of cells. The intraprostate tumor, although initially confined to the injected lobe, grew to involve the surrounding tissues and eventually metastasized to the lymph nodes and lungs. Occasional metastatic lesions were found in other organs also. Acid phosphatase could be domonstrated by histochemical staining of frozen tumor sections, and elevated levels of the enzyme were seen in the sera of rats bearing long-term subcutaneous tumors. During investigation of the tumor, a fast-growing tumor line arose that grew equally as well in females as in males. The histology of this tumor was that of an undifferentiated anaplastic tumor.


Tumors of the heart and stomach induced in European hamsters by intravenous administration of N-methyl-N-nitrosourea. One hundred twenty captured European hamsters (Cricetus cricetus L.) were inoculated iv with 1/5, 1/10, or 1/20 the median lethal dose of N-methyl-N-nitrosourea (MNU) once weekly for 18 weeks. MNU induced mainly sarcomas in the heart and squamous cell carcinomas and sarcomas in the stomach. The highest incidence of such tumors was demonstrated by the lowest-dosage group. They were more frequent in males than in females. The effect of MNU was also localized at the site of administration and resulted in squamous cell carcinomas of the oral cavity. The possible fibroblastic or neurogenic origin of the sarcomas was explored.


Computer-assisted collection and analysis of pathology data. At the National Center for Toxicological Research, long-term experiments with mice requiring the collection, storage, retieval, and analysis of an inordinate amount of data necessitated the development of an automated pathology information system. The system included automatic collection of mouse body and organ weights and hematologic findings; the use of mark-sensitive forms for the collection of gross and microscopic findings; and the use of computers for the storage, retrieval, and analysis of pathology data.


Markers of neoplastic transformation in epithelial cell lines derived from human carcinomas. Density-dependent inhibition of growth, plating efficiency on confluent monolayers of 3T3 cells, and growth in agar have been measured in epithelial tumor cell lines to determine whether they have properties in common with transformed mesenchymal cells. Five lines (RT4, RT112, J82, T24, and EJ) were derived from different human bladder tumors and HT29 was from a human colon tumor. All the lines resembled transformed "fibroblasts" in the absence of density-dependent inhibition of growth and the cell-surface large external transformation-sensitive protein, and they could form colonies on 3T3 monolayers. Only RT112, EJ, and HT29 were tumorigenic in nude mice, and the tumors had many of the structural and ultrastructural features of both the original tumor and the tissue of origin, even though the cells had been through many in vitro passages. Four of the lines (J82, T24, EJ, HT29) grew in agar, so that in some cell lines no correlation of growth in agar with tumorigenicity in nude mice was found: J82 and T24 were nontumorigenic and grew in agar, whereas tumorigenic line RT112 did not grow in agar. The ability to grow in agar did not appear associated with the production of high levels of plasminogen activators.


Effect of Corynebacterium parvum, methanol-extraction residue of BCG, and levamisole on macrophage random migration, chemotaxis, and pinocytosis. Three parameters of macrophage function: random migration, chemotaxis, and pinocytosis, were studied in the guinea pig after administration of Corynebacterium parvum, methanol-extraction residue of BCG, and levamisole (LMS), a synthetic anthelmintic. Macrophage migration studies were performed with a modified Boyden chamber. Pinocytosis was assessed by the uptake of colloidal 198Au. After ip administration, each of the three immunostimulators induced an increase in macrophage chemotactic responsiveness and, to a lesser extent and duration, in random motility. Kinetic, dose-response, and time course data for the effect of each agent on macrophage movement were explored. LMS was the most effective stimulator of macrophage activation, which occurred earlier and persisted longer than it did with the other agents. Macrophages from animals receiving each of the agents showed enhanced pinocytosis. Measurement of macrophage random migration, chemotaxis, and pinocytosis appeared to provide a rapid and quantitative assessment of several parameters of macrophage function and, when studied with other immunologic parameters, may provide useful tools for the evaluation of potential immunoadjuvants.


Effect of age on nodule induction by azaserine and DNA synthesis in rat pancreas. The efficacy of various azaserine treatment durations was evaluated with respect to induction of atypical acinar cell nodules in Wistar rat pancreas and was related to animal age and rate of pancreatic DNA synthesis during growth. The sensitivity to nodule induction was maximal in postnatal rats when the rate of pancreatic DNA synthesis was high, whereas treatment of weanlings was less effective and treatment of mature rats was least effective. When weaned growing rats were given 1, 3, or 5 weekly injections of 30 mg azaserine/kg, the number of nodules induced was proportional to the number of injections. A single dose at this level did not induce detectable pancreatic necrosis or inflammation; therefore, DNA synthesis due to regeneration was probably not a major factor in the initiation of nodules. We concluded that multiple daily injections of [3H]thymidine during the first or second postnatal week provided DNA of sufficiently high specific activity for use in DNA repair and biochemical toxicity studies.


Esophageal and gastric cancers with metastases induced in dogs by N-ethyl-N'-nitro-N-nitrosoguanidine. Three 6-month-old male beagle dogs were given a solution of 150 microng N-ethyl-N'-nitrosoguanidine (ENNG)/ml to drink ad libitum for 9 months. They all developed esophageal squamous cell carcinomas and gastric adenocarcinomas. The stomach adenocarcinomas were mostly in the antrum along the lesser curvature and were either well differentiated or poorly differentiated, with or without signet ring cells. The well-differentiated adenocarcinomas metastasized to the liver, and the poorly differentiated ones metastasized to the lymph nodes. The gastric mucosa in the antrum was atrophic, and the muscularis mucosae was fibrotic. Esophageal lesions were multicentric moderately differentiated squamous cell carcinomas, and they developed without diffuse hyperplastic changes of the epithelium. One dog with a large ulcerated carcinoma of the esophagus had metastases in the lung, liver, peritoneum, and abdominal lymph nodes. One dog also had a hemangiosarcoma with hepatic metastasis and spindle cell sarcoma in the stomach and duodenum, respectively.


Two syngeneic cell lines from human breast tissue: the aneuploid mammary epithelial (Hs578T) and the diploid myoepithelial (Hs578Bst) cell lines. We characterized two human cell lines (Hs578T and Hs578Bst), which provide several unique features that should be useful in the study of breast disease. Hs578T, derived from a carcinosarcoma, is epithelial in origin. Hs578Bst, established from normal tissue peripheral to the tumor, is myoepithelial in origin. This is the first report of companion cell lines, one malignant and one normal, established from the same organ.


Regression of bovine ocular carcinoma by treatment with a mycobacterial vaccine. Hereford cows with naturally occurring ocular squamous cell carcinoma were treated by injection of BCG cell-wall vaccine into the tumor. Regression or arrest of disease was observed in 71% of treated animals. The disease progressed in all untreated animals and animals treated with improperly compounded vaccine. At autopsy, most animals with progressive disease had lymph node metastases.


Immune response to L2 guinea pig leukemia-specific antigens. Strain 2 guinea pigs were immunized with the LE-L2C cell line and challenged with either LE-L2C or BZ-L2C, a subline of L2C leukemia deficient in la gene product and C3 receptor. The LE-L2C-immune animals were completely protected from a challenge of 5 X 10(6) cells (100 times the lethal dose) of either cell line, and the delayed skin test responses for both lines of injected cells were equivalent. Thus established in vivo immune recognition of the leukemia antigen was similar for both cell lines. After repeated immunization of strain 2 animals with BZ-L2C cells, animals were challenged with 3 X 10(5) viable cells; 9 of 13 animals survived. A subsequent 5 x 10 (6) challenge lowered survival to 8 of the 13 animals. The skin test response to BZ-L2C developed slowly during the immunization period, but survivors of the viable cell challenge exhibited good responses to skin testing with BZ-L2C or LE-L2C leukemic cells. Thus the BZ-L2C cell line possesses a leukemia-specific antigen, but the immunogenicity of this mutant line is decreased when compared to that of the LE-L2C line.


Suppression of dibutylnitrosamine-induced bladder carcinomas in hamsters by dietary indole. Effect of dietary indole on the urinary bladder tumorigenesis by chronic dibutylnitrosamine (DBN) treatment was evaluated in hamsters. In the first experiment, in which DBN-water and diet were given ad libitum, dietary indole significantly suppressed bladder tumor incidence. The inhibitory effect was more pronounced in males. In  the second experiment, in which consumption of both diet and DBN-water was rigidly controlled by pair-feeding, dietary indole again significantly suppressed bladder tumor incidence; its effect was similar in both males and females. This suppressive effect of indole on bladder tumorigenesis contrasted markedly with its failure to suppress tumors at other sites such as nasal sinuses, trachea, esophagus, and fore-stomach.


Cocarcinogenic interaction between D,L-tryptophan and 4-aminobiphenyl or 2-naphthylamine in dogs. Administration of a dietary supplement of 6 g D,L tryptophan/day  for 4 1/2 years following the administration of a single dose of 50 mg 4-aminobiphenyl/kg produced a bladder tumor in 1 of 4 beagle dogs. No tumors were observed in 6 dogs given the same dose of 4-aminobiphenyl without supplemental tryptophan. In a second experiment, administration of a supplement of 6 g D,L-tryptophan/day for 3 years following the administration of 5 mg 2-naphthylamine/kg/day for 30 days produced bladder tumors in 2 of 4 dogs. No tumors or other bladder pathology was produced by treatment of 4 dogs with this dose of 2-naphthylamine alone. Dogs given D,L-tryptophan alone developed no bladder tumors, but in most dogs receiving tryptophan the "tryptophan effect", i.e., a darkly stained mucosa with white areas of focal hyperplasia, was observed. Both experiments suggest a role of D,L-tryptophan as a cocarcinogen or promotor in the induction of bladder cancer.


Effect of intestinal microflora suppression on liver carcinogenicity of dimethylnitrosamine in Syrian hamsters. Liver tumors induced by dimethylnitrosamine showed an increased incidence in Syrian hamsters with intestinal microflora suppressed by antibiotics, as compared with the incidence in animals not treated with antibiotics. The possible mechanisms, including metabolic alterations caused by changes in the normal microflora population, were discussed.


Origin in tobacco smoke of N'-nitrosonornicotine, a tobacco-specific carcinogen. To evaluate risk factors and to approach methods of reduction of the carcinogenic potential of cigarette smoke, the transfer rate of N-nitrosonornicotine in a popular U.S. blended cigarette into mainstream smoke was quantitatively determined. The mean transfer rate was 11.3%; thus approximately 46% of the tobacco-specific carcinogen in the smoke came from the tobacco, and the remainder was synthesized during smoking.


Simplified radioimmunoassay for viral antigens: use of Staphylococcus aureus as an adsorbent for antigen-antibody complexes. In a rapid method for the radioimmunoassay (RIA) of viral antigens, Staphylococcus aureus was used as the adsorbent for antigen-antibody complexes ("protein A" molecules on the cell walls of certain strains of staphylococci have a strong affinity toward IgG molecules). The results showed that this method could be used instead of the double-antibody technique, with the same or probably higher sensitivity in precipitation as well as competition RIA's.


Spontaneous fibrosarcoma in a thymus-transplanted nude mouse. A fibrosarcoma was observed in a 6-month-old nude mouse that had a neonatal thymus transplant and had been kept since birth under nearly pathogen-free conditions. This was the first tumor found in more than 1,000 nude mice maintained in a colony in which mammary carcinomas were observed among the female heterozygous breeders. This finding confirmed the fact that malignant neoplasms are rare in an animal model that lacks T-cell-mediated immunologic capability and apparently contradicted the postulates of the immunosurveillance theory.


Transhepatic cholangiography.

Transhepatic cholangiography. Transhepatic cholangiograms were done on 81 patients. The Teflon sheath needle was used for 71 cases and the "Chiba" needle for ten. The "Chiba" needle transhepatic cholangiogram is recommended for the study of jaundiced patients with biliary duct dilatation, which is detected by ultrasound or computerized axial tomography.


Failure of selective pituitary adenomectomy to cure acromegaly. Complete selective pituitary adenomectomy for acromegaly may fail to cure the underlying endocrinopathy. A 40-year-old man who had acromegaly of ten-years duration was initially treated with pituitary irradiation eight years prior to surgery. Three years after irradiation his mean GH concentration decreased from 53 to 9.0 ng/ml, then subsequently increased, and was associated with development of severe hyperglycemia. A transsphenoidal microdissection was performed, and a discrete pituitary adenoma was visualized and totally removed at surgery. GH concentrations decreased from 46 to 5.0 ng/ml one week after surgery and to 3.0 ng/ml 10 months after surgery. Despite improvement postoperatively, the patient still demonstrated abnormal GH dynamics and responses to the GTT, abnormal nocturnal and random GH sampling, and abnormal responses to the L-dopa, apomorphine, and TRH stimulation tests. Acromegaly could be caused by either hypothalamic hyperfunction, an autonomous pituitary tumor, or both; but the latter would not explain the results obtained in this patient. This suggests that reduction of GH to normal concentrations following selective pituitary adenomectomy is not synonomous with curing acromegaly.


Evaluation and treatment of patients with dementia. Dementia is a common problem in middle-aged and elderly adults. Thorough diagnostic evaluation uncovers a significant minority who are suffering from an underlying neuropsychiatric disease amenable to specific medical or surgical therapy. Even in those patients with brain atrophy of unknown cause the physician can offer counseling and environmental manipulation to maximize remaining intellectual capacity.


The CVP catheter: an invasive therapeutic adjunct. Invasive techniques continue to be used in monitoring and treating the critically ill patient in spite of preference for, and increasing use of, non-invasive or minimally invasive techniques. With or without simultaneous determination of left ventricular filling pressure, the central venous pressure (CVP) catheter is widely used for gaining access to the right heart, determining right ventricular filling pressure, and for rapid infusion of fluids or medications. Even in expert hands the insertion of the CVP line is occasionally followed by complications that may require therapeutic interventions of varying degrees of severity. The possible hazards from this simple and often necessary therapeutic adjunct call for a systematic informed-consent approach which is often neglected despite its relevance in today's clinical practice. The data presented are typical of a standard teaching hospital experience. Securing informed consent for CVP catheter insertion is encouraged.


Pattern of organ involvement in Burkitt's lymphoma in Ibadan: a review. A retrospective study was conducted on 127 patients with Burkitt's lymphoma who had partial or complete records of both clinical and pathologic features of their disease. An analysis and description of the anatomical tumor distribution based on post-mortem examinations was made, highlighting lack of correlation between clinical staging and pathologic findings. A consistent observation was the high incidence of involvement of the kidney, liver, and heart in each of the stages (I-III).The data to be presented will clarify at least in part, the unpredictability of response of Burkitt's lymphoma patients to chemotherapeutic agents, because total cell-kill, needed for cure and/or prevention of relapse, is made almost impossible due to the widespread tumor volume carried by the patients.Effective supportive treatment might be a life-saving adjuvant regimen in treatment of Burkitt's lymphoma patients, where the organs so commonly involved might fail to function.


Pruritus in sickle cell disease: response to cholestyramine. Even in the absence of gallstones, the hepatobiliary system should be suspected as a possible cause of pruritus in a patient with sickle cell disease. A case is presented in which a sickle cell patient had severe pruritus relieved by cholestyramine therapy.


A comprehensive program for meeting psychosocial needs of sickle cell anemia patients. Some of the methods used at the Comprehensive Sickle Cell Center at Los Angeles County-University of Southern California Medical Center are explored. Additional methods must be tried and evaluated with appropriate documentation. A determination of the true effectiveness of the methods used is a major challenge and provides opportunity for further investigation. Certainly the tasks are myriad. Paraphrasing Dr. Marilyn Gaston, "Through effective coordination and communication by a multi-faceted team, improvement of the quality of life with a realistic outlook for the future can be achieved."(1)


Race-related differences in peripheral blood and in bone marrow cell populations of American black and American white infants. The present study confirms that in healthy infants, a racial difference exists in peripheral blood hemoglobin levels, total leukocyte, and total neutrophil counts. Racial differences in the composition of the bone marrow are negligible. The only demonstrable difference is confined to the erythroid cell population. No significant difference exists in the incidence of myeloid cells or small lymphocytes.


A model for teaching health care professionals the components of the family. With the inclusion of the behavioral sciences component in the National Board Examination (1972), many medical schools have revised their curricula to include sociology in the training of the future physician. In dentistry, the behavioral sciences group of the International Association for Dental Research has increased in membership over the past years. In nursing, courses such as sociology, psychology, and anthropology have already been included in the curricula of various schools. However, the tremendous shortage of qualified social scientists who are capable of relating the behavioral sciences to the health care institution makes it difficult to meet the needs of graduate and professional schools everywhere. In view of this problem, this article will present a simple conceptual model that can be used as an organizational device for teaching the components of the family in relation to the health care institution.


Health manpower-planning without objectives. Health manpower developments of the past decade have resulted in an absolute increase in the number of health personnel, the expansion of the roles of some traditional categories of personnel, and the introduction of new professional categories. Inherent in these developments has been the acceptance of the principal that the relative and absolute increase in manpower would result in an increased availability of health services. Unfortunately, in the last decade, the correlation between increased numbers and increased services is not a strong one. The failure to link manpower needs to specific service objectives and to identify appropriate rates of substitution among professional types has resulted in a wastage of funds and energies.A framework for future planning must now be developed which (1) defines service priorities, (2) delineates the functions required to deliver those services, and (3) defines appropriate manpower categories with their rates of substitution to perform those functions. Training programs must be coordinated to allow appropriate linkages among categorical types of personnel. The maldistribution of health care service must be viewed as a result of the demographic maldistribution among the health professions as well as the maldistribution of organizational and financial incentives for provision of priority services as well as utilization of priority services.


Retarded development of fetal renal alkaline phosphatase in mice given 2,4,5-trichlorophenoxyacetic acid. Histologic study of the fetal offspring of maternal mice given 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) suggested that the previously reported fetal "cystic kidneys" were due to a retardation in fetal renal development and downgrowth of the renal papilla into the pelvis. To determine a possible retardation in renal alkaline phosphatase or functional development, maternal mice received by gavage 60-120 mg/kg, 2,4,5-T on days 6-14 of pregnancy. At necropsy on day 17, the fetal kidneys were excised and fixed 24 hr in cold 65% ethanol. Paraffin sections stained by Gomori's method revealed alkaline phosphatase mainly in tubules in the inner renal cortex. Fetal kidneys showing diminished or no alkaline phosphatase were designated subnormal. There was a statistically significant greater incidence of subnormal fetal kidneys in the 2,4,5-T-treated mice than in the untreated controls. In three experiments, some mice were also sacrificed on day 18, and the incidence of subnormal fetal kidneys was significantly lower than on day 17. This retardation in renal alkaline phosphatase development indicates a retardation in renal functional development and indirectly supports the view that 2,4,5-T also retards the morphological development of the fetal kidney and is not a renal teratogen in mice. It also illustrates that selected histochemical studies may be helpful in a teratologic investigation.


Halothane in tissue of rats following repeated subanesthetic exposures. Halothane in tissues of the rat has been measured as a function of time after five successive 3 hr exposures to 670 ppm halothane. In epididymal and perirenal fat, halothane concentrations diminished rapidly from 310 ppm to 1-2 ppm in 24 hr, with trace amounts detectable 5 days later. Brain and liver levels were reduced from 16 and 9 ppm to less than 1 ppm within 3 hr. Blood halothane was reduced from 3 ppm to less than 0.5 ppm in the same time.


Effect of nitric oxide on the oxygen transport of human erythrocytes. The oxygen dissociation curve of partially NO-liganded hemoglobin of human erythrocytes is measured. As the percentage of NO ligation increases, the affinity of nonliganded hemoglobin for oxygen increases and the heme-heme interaction decreases; furthermore, methemoglobin is formed. Therefore, NO affects the oxygen transport function of hemoglobin, decreasing the oxygen supply to peripheral tissues, because of (1) simple diminution of the available hemoglobin by the tightly bound NO, (2) the high affinity of hemoglobin for oxygen, and (3) the inevitable formation of methemoglobin.


Methylmercury: effect on serum enzymes and humoral antibody. Dosages of 20 and 10 ppm methylmercury were toxic to rabbits while 1 ppm did not produce clinical signs or death. Serum alkaline phosphatase levels were elevated in all rabbits exposed to methylmercury. Methylmercury-exposed rabbits challenged to A/PR8 influenza virus had hemagglutination inhibition titers as much as four times lower than those of controls. Histopathologic lesions were found in the cerebellum of rabbits that died. The most significant features of this study were that methylmercury chloride suppressed the humoral immune system and resulted in increased serum alkaline phosphatase levels, which may aid in diagnosis when methylmercury poisoning is suspected.


Cytogenetic investigations on lymphocytes from workers exposed to vinyl chloride. Eleven male workers from the polymerization department of a vinyl chloride factory, seven people employed in the laboratory of another vinyl chloride plant, and ten controls from outside the factory environment were examined for the presence of chromosome aberrations in blood lymphocytes. Chromosome anomalies such as fragments, rings, translocations, and dicentrics were observed in most of the workers from the polymerization department. However, the medical history of these workers shows that they received frequent radiographs of the hands, feet, vertebral column, and digestive tract. It is therefore impossible to determine whether these chromosome anomalies result from vinyl chloride exposure.


Hepatic mixed-function oxidase activity in mice treated with methylated benzenes and methylated naphthalenes. Eight methylated benzene and five methylated naphthalene compounds were injected ip into adult male mice daily for 3 days. On the fourth day livers from control and treated mice were compared with regard to weight, microsomal N- and O-demethylase activity, and various spectral characteristics of microsomal cytochrome P-450. Three methylated benzenes and one methylated naphthalene produced significant increases in liver weight/body weight ratios, O-demethylation of p-nitroanisole, N-demethylation of aminopyrene, or a combination of these.


Thermodynamics and toxicity of sulfuric acid mists. The hygroscopic nature of sulfuric acid mist has not been totally appreciated in previous toxicological work. Sulfuric acid mist, by adsorption or desorption of water, equilibrates rapidly with the relative humidity of its environment. The measured particle diameters of a given sulfuric acid mist will increase in size as the particles adsorb water on entering the respiratory tract. For a dry climate of 5% relative humidity, sulfuric acid mist particles will triple in size in the respiratory tract. At 60% relative humidity, particles will double in size, while particles in humid regions will increase very little. In the respiratory tract, the particles will deposit according to their size at 98% relative humidity. In order to compare toxicity data for sulfuric acid mist, relative humidity must be carefully recorded throughout the experimental procedure.


Toxic effects of chlorinated cake flour in rats. Four experiments were conducted using weanling Wistar rats to determine whether chlorinated cake flour or its constituents were toxic. Levels of 0.2 and 1.0% chlorine added to unbleached cake flour significantly (p less than 0.01) reduced growth rate by 20.7 and 85.2% and increased liver weight relative to body weight by 16.7 and 25.3%, respectively. Lipids extracted from flour chlorinated at the same levels had similar effects. Rat chow diets containing 0.2 and 0.6% chlorine in the form of chlorinated wheat gluten reduced growth rate and increased liver weight as a percentage of body weight. A rat chow diet containing 0.2% chlorine as chlorinated flour lipids increased absolute liver weight by 40%, kidney by 20%, and heart by 10% compared to pair-fed controls.


Stimulation of DNA synthesis in the lungs of hamsters exposed intermittently to nitrogen dioxide. Stimulation of [3H] thymidine incorporation in the lungs of hamsters exposed singly and repeatedly to 10 ppm NO2 was studied. Within 24 hr of the first exposure to NO2, the response was characterized by a marked increase in [3H] thymidine labeling in the bronchi, bronchioles, and alveolar ducts, but not in the trachea or the peripheral alveoli. If, after the first NO2 exposure, subsequent exposures were repeated daily, no further stimulation of [3H] thymidine incorporation was observed in any part of the respiratory tract. However, if the interval between the first and subsequent exposures was 2-3 days rather than 1 day, stimulation of [3H] thymidine incorporation was observed in the bronchi, bronchioles, and alveolar ducts for up to 21 NO2 exposures. A significantly greater increase in [3H] thymidine incorporation was observed when the interval between subsequent exposures was extended to 7 days. However, no repeated exposure, whether at 2-3 or 7 day intervals, was as effective as the first NO2 exposure in stimulating incorporation of [3H] thymidine.


Tartrazine and the developing nervous system of rats. Rat dams were exposed to the artificial food color tartrazine (FD&C Yellow no. 5) at dietary levels of 0, 1, and 2% during gestation and lactation. The experimental offspring were continued on the same diets for approximately 3 months after weaning. No adverse physical or behavioral effects were noted in the dams. Fetal development and postnatal viability of the offspring were also normal. The only effect on postnatal development of the central nervous system (CNS) was a small transient change in neuromotor clinging ability of female offspring. The limited effect of tartrazine on the CNS was further evidenced by the facts that (1) the neurobehavioral profiles of the experimental weanlings revealed no significant abnormalities, and (2) morphochemical analysis of brain tissue, as well as brain weights, revealed no abnormalities. Tartrazine did appear to exert more general signs of toxicity in the offspring--namely, depressed body weight, an apparent reduction in thymus weight, and a slight elevation of red blood cells and hemoglobin.


Styrene and related hydrocarbons in subcutaneous fat from polymerization workers. Subcutaneous fat samples from 25 workers in a styrene polymerization plant have been analyzed for styrene, ethyl benzene, toluene, benzene, and benzaldehyde by gas chromatography. Styrene was found in samples from 13 of 17 workers who had been heavily exposed within the previous 3 days, 5 of 13 having been exposed 2-3 days earlier. Six workers 4-90 days removed from exposure and two 2-3 days removed from low (less than 5 ppm) exposures had no detectable styrene in fat tissue samples. Toluene and ethyl benzene were found in varying amounts in many samples, and benzene was observed in three samples. Benzaldehyde was observed at levels of 5-53 microng/g in all samples. Although urinary metabolites and breath levels of styrene are reported to be detectable for less than 24 hr following exposure, styrene was found in subcutaneous fat from the subjects of this study for as long as 3 days after the most recent occupational exposure. The combination exposures in such a setting are reflected in the variety of hydrocarbons found in fat samples of workers.


Detection of cardiac disturbances following thoracic trauma with high-frequency analysis of the electrocardiogram. High-frequency electrocardiograms (HF-ECG) were recorded from rabbits before and after delivery of blunt, nonpenetrating thoracic trauma. The conventional electrocardiographic record showed transient changes following the blow; the HF-ECG showed distinct notching and slurring in the QRS complex in most cases, which persisted throughout the course of the experiment. At autopsy, a contusive lesion of the right ventricle was always found.


Intermediate-degree blunt renal trauma.

Intermediate-degree blunt renal trauma. One hundred four cases of blunt renal trauma have been reviewed and analyzed, yielding 71 minor, 13 major, and 20 intermediate-degree injuries. Of the latter group, urinary extravasation (60%) and parenchymal laceration (100%) appear to be the dominant features exhibited by conventional radiographic maneuvers, neither of which suggest a de novo need for operation or interference with spontaneous resolution. Nephrectomy (9) and heminephrectomy (2) may often have been performed unnecessarily in this group, since nine such patients managed either nonoperatively (7), or undergoing negligible operative procedures (2) responded with total functional and anatomic restoration. Arteriography is endoresed as an important contributor of information encouraging conficdence in the conservative management of such cases.


Effects of enviromental temperature and femoral fracture on wound healing in rats. Femoral fracture, unilateral and bilateral, impaired the healing of dorsal skin incisions and formation of reparative granulation tissue in subcutaneously implanted polyvinyl alcohol sponges judged histologically and by breaking strengths and hydroxyproline contents, respectively, 1 week after injury in pair-fed rats kept at 22 degrees C. When rats were transferred to a room at 30 degrees C immediately after skin incision and sponge implants, with or without unilateral fracture, no differences in healing were observed between the two groups. Rats with skin incision, sponge implants, and either femoral fracture or sham-fracture excreted more urinary nitrogen than preoperatively when kept at 22 degrees. Counterpart groups transferred to a 30 degrees room right after operation excreted less urinary nitrogen than preoperatively, but because of lower food intakes postoperatively, the ratio of urinary nitrogen to food intake nitrogen was increased. With equivalent food intakes, pair-fed rats with fracture kept at 22 degrees postoperatively lost more weight and excreted more nitrogen than corresponding rats transfered to a 30 degrees room.


Effects of testosterone propionate and environmental temperature on nitrogen balance and wound healing of rats with and without femoral fracture. Testosterone exerted a sparing effect on nitrogen urinary excretion and balance in pair-fed rats with femoral fracture, dorsal skin incision, and subcutaneously implanted polyvinyl alcohol sponges, whether kept at 22 or 30 degrees C for 1 week postoperatively. No difference in 14-or 21-day wounds was observed among rats with dorsal skin incision and sponge implants, kept at 22 degrees, and starting immediately after operation, given daily IM injections of either 3 mg or 1 mg testosterone propionate in peanut oil, compared with similarly wounded rats given saline or peanut oil injections. Wound healing was impaired in rats kept at 22 degrees for 7 days following a skin incision, sponge implants, and femoral fracture. Testosterone administration begun immediately after operation, 2 mg/day subcutaneously, did not improve the impaired healing of pair-fed rats with fracture compared with rats without fracture. There was somewhat less collagen and a higher ratio of ground substance to collagen in the skin incisions and sponge granulomas of the testosterone group. No differences in wound healing were observed in pair-fed rats moved to a 30 degrees room postoperatively, among rats with or without fracture, nor among rats with femoral fracture given testosterone or saline subcutaneously.


Renal ischemia and contrast urography.

Renal ischemia and contrast urography. Exposure to urographic contrast material, meglumine diatrizoate, in a previously ischemic kidney was investigated for its effect on renal function. Eleven mongrel dogs, 19.8-32.7 kg in weight, having the left kidney ischemic for 90 minutes were given either 4 cc/kg meglumine diatrizoate or control, Ringer's lactate solution. The effect on creatinine clearance of the left kidney was determined acutely and at 72 hours. Statistical analysis of the data reveals no deleterious effect from exposure to meglumine diatrizoate over and above the effect of the ischemic episode. There was no statistically significant effect on recovery of renal function in this animal model.


Changing concepts in the treatment of penetrating cardiac injuries. Thirty consecutive patients with penetrating wounds to the heart underwent surgery at the Medical College of Virginia Hospital. Classical signs of tamponade usually were not present. A precordial wound, positive pericardiocentesis, and shock were the most common indications of cardiac injury. Immediate operation is recommended for all penetrating injuries; pericardiocentesis is employed only for a diagnostic tool and for emergency relief of tamponade.


Metabolic and clinical considerations in an unusual patient with 35% BSA deep partial-thickness thermal injury. The treatment of a moderately large burn usually require blood and its products to restore circulating blood volume, achieve metabolic stabilization, and promote wound healing. This case study demonstrates the remarkable metabolic response to injury in a patient refusing blood products administration. Specific management considerations are discussed.


One-stage debridement and plastic repair of compound comminuted depressed skull fractures with methyl methacrylate. A one-stage technique is described for repair and debridement of compound, comminuted depressed skull fractures using methyl methacrylate, which appears to be simple, effective, and safe, provided that the paranasal sinuses are not involved. If so, a two-stage debridement and repair are recommended. In the series reported, of 25 patients, only one developed infection, which was successfully treated with further operative repair and antibiotics.


Splenic injury due to indirect torsion.

Splenic injury due to indirect torsion. A patient with subcapsular rupture of the spleen in association with indirect torsion trauma is reported. The spleen was surrounded by bands and adherent organs. The injury was considered to have been caused by a stretched band. In patients with hemoperitoneum, celiac angiography is advocated for diagnosis when immediate surgery is not indicated.


Traumatic aorto-caval-portal-duodenal fistula: case report. A 19-year-old male was admitted 5 days following a stab wound of the epigatrium. An aorto-caval-portal-duodenal fistula was documented. The operative approach and recommendations for management are described.


Superior gluteal artery aneurysm.

Superior gluteal artery aneurysm. Aneurysms of the gluteal arteries are an uncommon complication of trauma to the gluteal region. A case report is presented following a gunshot wound to the buttock in a 36-year-old female. Diagnosis and management are discussed, with a review of the recent surgical literature.


Persistent post-traumatic pseudosheath formation secondary to a movable organized blood clot. Pseudosheath formation secondary to post-traumatic hematoma is described in a patient following blunt trauma to the lateral thigh. The theory of its formation and progression is discussed.


Seromucoids in chronic bowel disorders of tropics. Serum seromucoid estimation was carried out in 60 normal controls and 100 patients. (1) The serum seromucoid values in normal subjects ranged between 37-5-127 mg.% (69-35 +/- 17-84). (2) Serum seromucoid levels were found to be increased in patients of abdominal tuberculosis (210-95 +/- 86-49). These values corresponded directly to the severity of the disease. (3) Seromucoid levels came down to normal after three months of antitubercular treatment. (4) There was an inverse relationship between serum albumin and seromucoids. (5) Serum seromucoids were also found to be increased in patients of amoebiasis (157-54 +/- 37-61), with associated active colonic or hepatic disease. Asymptomatic cyst passers had normal levels of serum seromucoids. (6) Serum seromucoid values were within normal limits in patients with tropical sprue and irritable colon syndrome.


Co-existing post-stapedectomy fistula and acoustic neuroma. A 66 year old woman had light-headedness and ataxia which increased slowly despite tissue seal of a post-stapedectomy oval window fistula. The cause was shown to be a large acoustic neuroma, ipsilateral to the side of the fistula. The history, physical, and laboratory findings are presented.


Traumatic thermal injuries of the middle ear. Eleven cases of thermal otic injury are reviewed. The final outcome of therapy, whether medical or surgical is poor--only two of 11 patients were restored to normal. The possible pathogenesis of failure is discussed. Most cases could have been prevented by wearing ear plugs.


Malignant schwannoma of the parapharyngeal space. There is a variety of tumors which may occur in the parapharyngeal space; malignant Schwannoma is one of the rarer type. The authors present such a case and discuss the clinical and microscopic diagnosis. Treatment is wide surgical excision, as recurrence rates are high and the prognosis poor.


Epiglottic abscess: its occurrence and management. Twenty-six cases of epiglottic abscess from the English literature are reviewed. The condition occurs nearly exclusively in adults and it has a high mortality rate (30 per cent). The lingual surface of the epiglottis is most commonly involved and less than half of the patients present with severe airway obstruction. The management of the condition is dictated by the degree of airway obstruction.


Phycomycosis in an apparently normal host. A case presentation of phycomycosis recognized by histology but unproved by cultural methods is presented. The patient was successfully treated with Amphotericin B. Even though specific genus of phycomycosis was not identified, the clinical characteristics and apparently healthy host suggest the possibility of the fungus being of the order of entomophthorales. Classification, pathophysiology, various clinical types, diagnostic aids, and principles of treatment are discussed.


[Complications in rhinoseptoplasty].

[Complications in rhinoseptoplasty]. We have done a survey of our first 115 cases of rhinoseptoplasty. We noticed that 19 per cent of the patients presented partial nasal obstruction following first surgery, thus requiring a second surgical procedure. We attempt to suggest a few causes of this high percentage of failures and suggest some technical modifications to avoid such complications.


Fidelity of adenovirus RNA transcription in isolated HeLa cell nuclei. An in vitro nuclear system from adenovirus type 2-infected cells was developed to study transcription of viral RNA. Nuclei isolated from adenovirus-infected HeLa cells late in the infectious cycle synthesized in vitro only RNA from the r-strand of adenovirus DNA. Around 15% of the virus-specific RNA in isolated nuclei was polyadenylated. Short pulse labeling of nascent RNA followbd by hybrization of size-fractionated RNA to specific restriction endonuclease fragments of the genome suggested that the origin(s) for transcription is located on the r-strand in the left 30% of the adenovirus 2 genome at late times in the infectious cycle. Pulse-chase experiments were used to estimate the elongation rate of adenovirus high-molecular-weight RNA in isolated nuclei. An elongation of a least six nucleotides per second was observed in vitro. Viral RNA synthesis in the vitro nuclei showed several similarities to the in vivo system late in the infectious cycle.


Mitochondrial DNA synthesis in adenovirus type 2-infected HeLa cells. Mitochondrial DNA synthesis in adenovirus type 2-infected HeLa cells was measured at various times from 0 to 24 h postinfection. Although viral infection effectively turned off host chromosomal DNA synthesis, mitochondrial DNA synthesis was not inhibited. These findings indicate a dissociation between the regulation of host and mitochondrial DNA synthesis after infection with adenovirus type 2.


The segments of influenza viral mRNA.

The segments of influenza viral mRNA. Influenza viral mRNA, i.e., complementary RNA (cRNA), isolated from infected cells , was resolved into six different species by electrophoresis in 2.1% acrylamide gels containing 6 M urea. The cRNA's were grouped into three size classes: L (large), M (medium-size), and S (small). Similarly, when gels were sliced for analysis, the virion RNA (vRNA) also distributed into six peaks because the three largest vRNA segments were closely spaced and were resolved only when the gels were autoradiographed or stained. Because of their attached polyadenylic acid [poly(A)]sequences, the cRNA segments migrated more slowly than did the corresponding vRNA segments during gel electrophoresis. After removal of the poly(A) by RNase H, the cRNA and vRNA segments comigrated, indicating that they were approximately the same size. One of the cRNA segments, S2, was shown by annealing to contain the genetic information in the vRNA segment with which it comigrated, strongly suggesting that each cRNA segment was transcribed from the vRNA segment of the same size. In contrast to the vRNA segments, which when isolated from virions were present in approximately 1:1 molar ratios, the segments of the isolated cRNA were present in unequal amounts, with the segments M2 and S2 predominating, suggesting that different amounts of the cRNA segments were synthesized in the infected cell. The predominant cRNA segments, M2 and S2, and also the S1 segment, were active as mRNA's in wheat germ extracts. The M2 cRNA was the mRNA for the nucleocapsid protein; S1 for the membrane protein; and S2 for the nonstructural protein NS1.


Structural polypeptides of adenovirus type 16 incomplete particles. The polypeptides of adenovirus type 16 incomplete particles, with average buoyant densities of CsCl of 1.33, 1.318, 1.305, and 1.299 g/cm3 and DNA content of less than 1 U genome size, were compared with the polypeptides of the complete virion (density, 1.344 g/cm3, and 22 x 10(6) daltons of DNA) and late polypeptides in infected cells by using sodium dodecyl sulfate-gel electrophoresis. In agreement with other serotypes studied (types 2 and 3), the light particles lack polypeptides V, VI, and VII. In adenovirus type 16, eight other major polypeptides are found, with apparent molecular weights of 59,000 (59K), 46K, 31K, 30K, 28K, 27K, 26K, and 19K. The 30K to 31K and 27K to 28K polypeptides are phosphorylated. The 27K and 19K polypeptides are precursors, whereas the 31K, 30K, and 26K polypeptides are chase products in the cell, as are polypeptides VI, VII, and VIII. The 26K polypeptide is proposed to be an intermediate in processing since it disappears from the young virions upon chasing. Although chase products, the 31K and 30K polypeptides are only associated with particles having buoyant density lower than 1.318 g/cm3. Polypeptides V and VII are only present in particles containing more than one quarter of the genome. No trace of cell histones could be detected in the purified incomplete particles. A major constituent of the incomplete particles, the 46K polypeptide, was rapidly labeled in the cell, and loss of radioactivity from this band was detectable only after 7 to 18 h of chase.


Adenovirus type 2 coded single-stranded DNA binding protein: in vivo phosphorylation and modification. The adenovirus type 2-coded single-stranded DNA binding protein (DBP) was shown to be a phosphoprotein and to exist in at least two forms that differ in mobility by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After a 30-min pulse with [35S]methionine or 32PO4, 35S- or 32P-labeled DBP had a nominal molecular weight of 74,000 whereas after a 30-min label followed by a 20-h chase, 35S- and 32P-labeled DBP had a nominal molecular weight of 77,000. Both large and small forms of 35S- and 32P-labeled DBP bound to single-stranded DNA-cellulose columns and were eluted by 0.4 to 0.6 M NaCl; both forms also were immunoprecipitated by antiserum against adenovirus type 1-simian virus 40-induced tumor cells (this antiserum contains antibodies against DBP) and by monospecific antiserum against 95 to 99% purified DBP. With highly purified 32P-DBP labeled 7 to 10 h postinfection, it was shown that the 32P radioactivity was firmly associated with protein material (i.e., not contaminating nucleic acids or phospholipids) and had properties expected of a phosphoester of an amino acid; paper electrophoresis of acid hydrolysates of this preparation identified phosphoserine but not phosphothreonine. Phosphoserine but not phosphothreonine was also identified in acid hydrolysates of another preparation of 32P-DBP labeled for 30 min, chased for 20 h, and then immunoprecipitated by adenovirus type 1-simian virus 40 antiserum.


Precursors of the T4 internal peptides.

Precursors of the T4 internal peptides. The precursors of the two T4 internal peptides have been identified by in vitro cleavage of individual phage proteins eluted from sodium dodecyl sulfate-acrylamide gels. The precursor of internal peptide VII is p22, the product of T4 gene 22 and an essential component of the morphogenic core. The precursor of peptide II is a protein with a molecular weight of approximately 13,000, whose gene has yet to be defined by mutation. A newly detected protein of approximately 15,000 molecular weight is found to be cleaved and is, therefore, likely to be a component of precursor head structures.


Replication process of the parvovirus H-1. IX. Physical mapping studies of the H-1 genome. The cleavage map of H-1 replicative-form DNA to the bacterial restriction endonuclease EcoRI, HaeII, HaeIII, HindII, HindIII, and HpaII has been determined. The 5'-phosphoryl end of the viral strand is on the right end of the molecule at or near the replication origin. Evidence is presented for the presence of inverted self-complementary sequences at the right end that differ from those at the left end. These sequences allow a foldback of the DNA after denaturation, and a minority of the native replicative-form DNA has the foldback configuration. The possible role of these structures in H-1 DNA synthesis is discussed.


Structural aberrations in T-even bacteriophage. IX. Effect of mixed infection on the production of giant bacteriophage. To date, the production of T-even bacteriophage with giant heads has been achieved in two ways: (i) by use of canavanine-arginine treatment of Escherichia coli B cultures infected by wild-type bacteriophage (Cummings and Bolin, Bacteriol. Rev. 40:314-359, 1976; Cummings et al., Virology 54:245-261, 1973), which give a size distribution of giants that is phage specific (Cummings et al., Virology 54:245-261, 1973); and (ii) by infection with certain missense mutants of T4D gene 23 (Doermann et al., J. Virol. 12:374-385, 1973; ICN-UCLA Symposium on Molecular Biology, p. 243-285, 1973) or temperature-sensitive mutants of gene 24 (Aebi et al., J. Supramol. Struct. 2:253-275, 1974; Biljenga et al., J. Mol. Biol. 103:469-498, 1976). We now report the effect of mixed infection with several mutants of T4D on both the production and the size of giant bacteriophage. We found that gene 24 mutant is a critical partner for the production of giants. Infection using T4.24 mutants together with either T4.23 mutants, T4B+ or T6+ led to the formation of giants with heads 10- to 14-fold longer than normal-length heads. Infection with amber 24-bypass 24 double mutants of T4D led to the production of giants when gene 23 mutant was used to co-infect. Addition of canavanine to the co-infected cultures could alter the size distribution of giants, depending on which phage were used to coinfect. Gene 22 mutants had a modifying effect on these results. In the absence of canavanine co-infection with gene 22 mutants prevented the production of giants, and in the presence of canavanine giants of 1.5 to 5 head lengths were found. We have interpreted these results to mean that critical concentrations of gene products 22, 23, and 24 interact to control head length in T-even bacteriophage.


Distribution of "minor" nicks in bacteriophage T5 DNA. Bacteriophage T5 DNA can be released from the phage particle in such a way that one end of 5 to 10% of the DNA molecules remains attached to either the phage head or tail. Under partial denaturation conditions, the DNA preferentially denatures in the vicinity of a nick so that the nicks can be located relative to the end that remains attached to the phage head or tail. Two classes of nicks were found. "Major" nicks were those found in more than 20% of the molecules and were located at the same points along the DNA molecule as reported by others. "Minor" nicks were found in 5 to 10% of the molecules and often occurred at specific locations near a "major" nick.


Sequence analysis of the nicks and termini of bacteriophage T5 DNA. Bacteriophage T5 DNA, when isolated from mature phage particles, contains several nicks in one of the two strands. The 5'-terminal nucleotides at the nicks were labeled with polynucleotide kinase and [gamma-32P]ATP, and the 3'-terminal nucleotides were labeled with Escherichia coli DNA polymerase I and [alpha-32P]dGTP. The sequences around the nicks were analyzed by partial nuclease digestion followed by homochromatography fractionation of the resulting oligonucleotides. The nicks had at least the sequence -PuOH pGpCpGpC- in common. In addition, the two 5' external termini had the first seven nucleotides in common.


Adenovirus type 2 nuclear RNA accumulating during productive infection. The viral-specific nuclear RNA which accumulates early and late during productive infection of HeLa cells by adenovirus-type 2 (Ad2) has been characterized with respect to its size and stability after denaturation by Me2SO. Early nuclear transcripts, under nondenaturing conditions, sediment in the range 28 to 45S, but treatment with Me2SO prior to sedimentation results in a shift to about 20S. Later nuclear RNA accumulates as a composite of two populations of molecules: one with a broad size distribution centering on 45S under nondenaturing conditions and less than 32S after denaturation and a second having a narrow size distribution around 35S which is quite stable to Me2SO. Analysis of late RNA by hybridization to Sma fragments of Ad2 DNA suggests that the 35S RNA species is derived from a limited portion of the left half of the viral genome.


Tyzzer's disease in wild-trapped muskrats in British Columbia. A diagnosis of Tyzzer's disease (Bacillus piliformis infection) was made in a group of recently-trapped muskrats (Ondatra zibethica). The major lesions consisted of enteritis and focal necrosis of the liver, with organisms resembling B. piliformis in hepatocytes on the periphery of these foci. The outbreak was associated with recent capture, housing, and the use of sulfaquinoxaline-medicated feed.


An epizootic of pneumonia in captive bighorn sheep infected with Muellerius sp. An epizootic of pneumonia in 20 captive Rocky Mountain bighorn sheep (Ovis canadensis) is described. The sheep were maintained in large paddocks for about 9 months after which, in the late summer, the entire herd died within a three week period. Large numbers of Muellerius adults, eggs and larvae were in the lungs of all sheep. First stage larvae were widely disseminated throughout the lungs and apparently elicited a granulomatous pneumonia. Pasteurella sp. and other bacteria were isolated from the lungs of several sheep but no Chlamydia, Mycoplasma or viral agents were recovered. Elevated serum fibrinogen levels and normal leukocyte values were found in blood samples taken from several sheep 7-14 days prior to death.


Helminth parasites of bighorn sheep in Oregon. The lungs and gastrointestinal tracts from 18 hunter-killed bighorn rams (Ovis canadensis californiana) were examined in total or in part for helminth parasites during a two-year study of three separate herds in Eastern Oregon. Prevalence was 100% with the lungworm Protostrongylus stilesi. The gastrointestinal fauna from 11 rams comprised Cooperia oncophora, Marshallagia marshalli, Nematodirus oiratianus, Oesophagostomum spp., Ostertagia occidentalis, O. ostertagi, Skrjabinema ovis, Trichostrongylus axei and Trichuris spp. Adult Wyominia tetoni and cysticerci of Taenia hydatigena were recovered from two of six livers examined. Additionally, searches for potential molluscan intermediate hosts for P. stilesi were conducted on one bighorn range. Snails identified as belonging to the genera Euconulus, Pupilla and Vallonia were found on both the summer and winter ranges.


Rabies virus and antibody in bats in Grenada and Trinidad. Rabies virus was detected by fluorescent-antibody and mouse inoculation tests in the brain of one bat, Artibeus jamaicensis, collected at La Tante, Grenada on 19 June 1974. No rabies virus was found in the brains and/or salivary glands of 411 other Grenadian bats of 6 species tested, including 56 A. jamaicensis. Rabies neutralizing antibody was detected by the rapid fluorescent focus inhibition test (RFFIT) in 27 of 353 Grenadian bats. Positives occurred in each of the 6 species sampled, with 40.5% prevalence in A. jamaicensis. In 11 of 86 Trinidadian bats of 4 species known to carry rabies, positive sera occurred only in A. jamaicensis (18.6%) and A. lituratus (18.1%). The potential use of the REFIT indetermining rabies activity is discussed.


Meningeal worm-induced neurologic disease in black-tailed deer. Neurologic disease attributable to Parelaphostrongylus tenuis was diagnosed in five black-tailed deer (Odocoileus hemionus columbianus) relocated from Oregon to Tennessee. Mortality occurred in the pre-release enclosure and in the release area. Infection with P. tenuis was considered the cause of an unsuccessful stocking attempt. In addition, neurologic disease was produced by experimental infection of a black-tailed x white-tailed deer hybrid. Clinical and pathologic findings were described for black-tailed and hybrid deer, and epizootiologic aspects of P. tenuis infections were discussed.