Lack of homology between dog and human placental alkaline phosphatases. Alkaline phosphatases [ALPases; orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] from dog and human placenta, liver, bone, kidney, and intestine were investigated by inhibition studies with L-homoarginine, L-phenylalanine, and L-phenylalanylglycyl-glycine; by thermostability studies; and by electrophoresis, both before and after treatment with neuraminidase. The inhibitions obtained for each inhibitor with dog placental ALPase closely match those obtained with dog and human liver, bone, and kidney ALPases, but are quite different from those obtained with human placental ALPase. Dog placental ALPase is thermolabile, as are dog and human liver, bone, and kidney ALPases, in marked contrast to human placental ALPase, which is very thermostable. Dog placental ALPase has the same electrophoretic mobility as dog liver, bone, and kidney ALPases after removal of sialic acid residues with neuraminidase. Desialated human placental ALPase differs electrophoretically from desialated human liver, bone, and kidney ALPases, which show the same mobilities. Dog and human intestinal ALPases are distinguished by these various criteria from the liver, bone, kidney, and placental ALPases of both species, but are similar to each other. These results suggest that the ALPase gene locus expressed in dog placenta is not homologous to that expressed in human placenta. Rather, it appears to be homologous to the ALPase locus expressed in dog and human liver and possibly also bone and kidney. Other incomplete data suggest that this may also be true for placental ALPase in other mammalian species. One possible explanation is that human placental ALPase, a relatively recent newcomer on the evolutionary scene, arose from a gene duplication that occurred subsequent to the evolutionary divergence of many other mammalian species.


Antigen-specific identification and cloning of hybridomas with a fluorescence-activated cell sorter. Myeloma-spleen cell hybrids (hybridomas) producing antibody to mouse immunoglobulin allotypes have been labeled with fluorescent microspheres coupled with myeloma protein antigens. The ratio of specific to nonspecific microsphere binding by viable hybridoma cells was about 100:1. By using a modified fluorescence-activated cell sorter (FACS), selected hybridoma cells in a mixture have been sorted individually into media in microculture wells, where, with thymocyte feeder cells, they developed into clones producing a desired monoclonal antibody. Viable cells were selected by measurement of their light scattering and autofluorescence properties. Rare antibody-producing clones were obtained without laborious screening and repeated subculturing. This technique should expand the range of monoclonal antibodies readily obtained from hybridomas and greatly facilitate the process of obtaining desired hybridomas.


Chemotactic factor for tumor cells derived from the C5a fragment of complement component C5. Previously, we have stablished that the fifth component of complement (C5) serves as an important source of mediators that have locomotory (chemotactic) activity for leukocytes and tumor cells. C5a, a fragment (Mr 11,200) derived from the NH2-terminal portion of the alpha chain of C5, is the major chemotactic peptide for leukocytes. The present studies demonstrate that cleavage of C5a with trypsin generates a derivative peptide that is chemotactic for tumor cells (Walker carcinosarcoma). This fragment has an estimated Mr of 6000 as assessed by gel filtration and does not require the COOH-terminal arginine of C5a, because equivalent amounts of chemotactic activity for tumor cells can be generated from des-Arg-C5a by digestion with trypsin. The C5a-derived chemotactic peptide for tumor cells demonstrates peak activity at approximately 1 pM. These studies emphasize the key role of the C5a region of the C5 molecule in the generation of peptides that affect locomotory responses of cells.


Biliary transport of IgA: role of secretory component. Biliary transport of rat immunoglobulin was studied by perfusion of isolated rat liver with blood containing radiolabeled immunoglobulin. Transport to bile was selective for polymeric IgA. Between 15 and 27% of polymeric IgA was transported from blood to bile during a 210-min perfusion period, and approximately 60% of the IgA transported to bile bore secretory component. Small quantities of IgM (0.12%) were transported; transport of IgG2 alpha, IgE, or monomeric IgA was not detected. Purification of radiolabeled polymeric IgA by affinity chromatography on human secretory component-Sepharose yielded a fraction that was transported more efficiently (i.e., up to 40% transported). In contrast, secretory IgA (colostral or biliary) was transported 1/25th to 1/12th as well as polymeric IgA myeloma protein. Complexes of 125I-labeled secretory component and polymeric IgA formed in vitro were transported poorly (0.1%) compared to polymeric IgA (26%). It was concluded that biliary transport of polymeric IgA requires combination of it with secretory component in the liver. In support of this hypothesis, rabbit IgG anti-rat secretory component antibodies were also transported to bile but normal rabbit IgG was not.


Protective effect of immunization with nonviral antigens against Friend leukemia virus in mice. DBA/2 mice immunized with poly(A).poly(U) complexed with methylated bovine serum albumin and emulsified in Freund's complete adjuvant were protected against challenge with Friend leukemia virus. There was no correlation between the level of antibody to the immunogen in the prechallenge serum and induced resistance to the virus. Although prechallenge sera of mice given the same amount of the duplex in a single inoculum bound 9.7% of poly(A).poly([(3)H]U) input, as compared to 45.3% bound by the prechallenge sera of mice given the immunogen in divided doses, both groups of mice were equally resistant to infection. Immunization with two other nonviral agents, bovine serum albumin fraction V or dinitrophenylated keyhole limpet hemocyanin, induced the same level of protection. A sparing effect of approximately 10(1.5) in infectivity was afforded the immunized mice. Immunization with either poly(A).poly(U) alone or with the carrier methylated bovine serum albumin was ineffective. In addition to antibodies to the respective immunogens, the prechallenge sera of the immunized mice also contained antibody to Friend leukemia virus gp71. The presence of such viral antibodies was not always related to resistance to infection by Friend virus. Some immunized mice that survived infection did not have gp71 antibody in their serum before challenge, and mice immunized with poly(A).poly(U) alone were susceptible to infection, although their prechallenge sera contained antibody to gp71. The mechanism involved in the induction of resistance to infection is not known. The effect may be mediated through a modification of the expression of both endogenous and exogenous type C viruses and affect immunological mechanisms controlling cellular responses.


Retrovirus sequences in a leukemic gibbon and its contact: evidence for partial provirus in the nonleukemic gibbon. Integrated viral DNA sequences were detected in tissues from two gibbon apes, a leukemic gibbon (6G-1) from whose leukocytes a distinct strain of gibbon ape leukemia virus (GaLVH) was isolated, and gibbon 6G-4, a contact of 6G-1 from the same colony that had uremia and cachexia of unknown origin. Although 6G-4 had no detectable neoplasia or viral proteins, its serum contained persistent antibody against GaLV antigens. Whereas DNA from most of the tissues of 6G-1 contained GaLV provirus, DNA from only three tissues (kidney, spleen, and liver) from 6G-4 showed detectable viral sequences, and the extent of hybridization in each case was lower than with 6G-1. After cleavage with BamHI, two virus-specific DNA fragments were detected in tissues of 6G-1. Only one of these fragments was detected in the positive tissues of 6G-4. The results indicate that: (i) 6G-4 was exposed to and infected by GaLV; (ii) early target sites for infection of gibbon by GaLV may be limited to a few tissues; and (iii) infection can be contained by integration of only partial provirus in a few tissues.


Measles and subacute sclerosing panencephalitis virus proteins: lack of antibodies to the M protein in patients with subacute sclerosing panencephalitis. Studies on the immunoprecipitation of the proteins of several different measles and subacute sclerosing panencephalitis (SSPE) virus strains from lysates of infected cells have revealed a relative lack of antibodies to the nonglycosylated viral membrane protein (M) in the sera of patients with SSPE, in contrast to a high level of antibodies to other viral proteins. No evidence was found for the existence of a large number of antigenically unique M proteins. In contrast to SSPE sera, other hyperimmune human or rabbit sera precipitated M protein as well as the other viral proteins. The results suggest that in patients with SSPE there is either diminished synthesis of M, or it is not recognized normally by the immune system, and that an abnormality in M protein is involved in the pathogenesis of SSPE. Although differences in electrophoretic mobility of the M proteins and several other viral proteins were observed among different SSPE and measles strains, there was no pattern characteristic of SSPE strains, nor could these strains be distinguished by peptide mapping.


Avermectin B1a irreversibly blocks postsynaptic potentials at the lobster neuromuscular junction by reducing muscle membrane resistance. Avermectin B1a, a macrocyclic lactone with broad spectrum anthelmintic activity, affects neuromuscular transmission in the lobster stretcher muscle. Perfusion of the muscle with 1-10 microgram of the drug per ml eliminates inhibitory postsynaptic potentials within a few minutes. Intracellularly recorded excitatory postsynaptic potentials are gradually reduced in amplitude over 20-30 min, and their falling phases become faster; there is no effect, however, on extracellularly recorded excitatory potentials. Avermectin B1a reduced the input resistance of the muscle fibers with a time course similar to that of the reduction of excitatory potentials. Washing for up to 2 hr with drug-free solution fails to reverse the drug's effects. However, perfusion with 20 microgram of picrotoxin per ml results in recovery of the excitatory potentials and input resistance. Avermectin B1a also blocks the firing of the crayfish stretch receptor neuron, and this block is also reversed by picrotoxin. We hypothesize that the reduction in excitatory postsynaptic potentials after avermectin B1a treatment is caused solely by reduction in membrane resistance; additional experiments suggest that the reduction in membrane resistance is due to the opening of membrane Cl- channels, perhaps including those regulated by gamma-aminobutyric acid at the inhibitory synapse.


Immunocytochemical localization of gamma-aminobutyric acid transaminase at cellular and ultrastructural levels. gamma-Aminobutyric acid transaminase (GABA-Tase; 4-aminobutyrate:2-oxaglutarate aminotransferase, EC 2.6.1.19) immunoreactivity in the rat's cerebellum was studied by light and electron microscopy with indirect immunofluorescence and peroxidase-antiperoxidase methods. Evidence is presented for neuronal and neuroglial compartments of GABA-Tase. Labeled neurons included stellate, basket, Purkinje, and Golgi cells of the cortex and a few large neurons in the deep nuclei. Labeled neuroglia included those surrounding Purkinje cells, their radial fibers in the molecular layer, and astrocytes in the granular layer and deep nuclei. No evidence for sagittal microzonation was found. At the ultrastructural level, GABA-Tase immunoreactive sites were localized to cell surface membranes, intracellular organelles, and the cytoplasmic matrix. GABA-Tase immunoreactivity at synapses could be localized precisely to pre- and postsynaptic membranes in gamma-aminobutyric acid (GABA)-containing as well as non-GABA-containing neurons. Specific label was absent from tissues treated with normal rabbit preimmune sera. GABA-Tase labeling was more intense in tissues from animals anesthetized with ether than with barbiturates and after formaldehyde fixation without glutaraldehyde. Increased GABA-Tase immunoreactivity was observed on treatment with colchicine, GABA with oxamic acid, GABA, harmaline, norepinephrine and glutamate, or diazepam (in order of decreasing effectiveness). Serotonin produced no detectable change, and apomorphine and muscimol decreased the immunoreactivity.


Inhibition of neuronal acetylcholine sensitivity by alpha-toxins from Bungarus multicinctus venom. Bungarus multicinctus venom contains several alpha-toxins in addition to the widely used alpha-bungarotoxin (Bgt 2.2). We have found that two of the alpha-toxins (Bgt 3.1 and 3.3) inhibit neuronal acetylcholine (AcCho) sensitivity when tested on ciliary ganglion neurons in cell culture. Over 90% of the AcCho sensitivity recorded in response to iontophoretic application of AcCho was blocked when the neurons were incubated with either of the toxins at 10(-7) M for 1 hr at 37 degrees C. The blockade could be partially reversed by incubating the neurons for 1-2 hr in medium lacking the toxins. The neurons also had a high-affinity binding site for Bgt 2.2, as indicated by binding studies with rhodamine-labeled Bgt 2.2. Concentrations of Bgt 2.2(10(-7) M) that should be nearly adequate to saturate the high-affinity site, however, had no detectable effect on AcCho sensitivity of the neurons. Higher concentrations of Bgt 2.2(10(-5) M) produced a partial inhibition of AcCho sensitivity, suggesting either that the neurons had two classes of binding sites for Bgt 2.2 (with the low-affinity site affecting AcCho sensitivity) or that the preparation of Bgt 2.2 contained minor components (e.g., Bgt 3.1 or 3.3) that were responsible for the blockade. The mechanisms by which Bgt 3.1 and 3.3 inhibit neuronal AcCho sensitivity remain unknown. If they bind specifically to the AcCho receptor, they will be useful agents for studying the distribution and regulation of this membrane component.


Opioid peptides and alpha-melanocyte-stimulating hormone in genetically obese (ob/ob) mice during development. Compared to littermate controls (C57BL/6J ob/?), body weights of genetically obese (ob/ob) mice are significantly higher at 1-6 months of age; the greatest percentage weight gain of the ob/ob group occurs during the first 3 months of life. Levels of pituitary immunoreactive beta-endorphin and immunoreactive alpha-melanocyte-stimulating hormone are also significantly elevated in ob/ob animals compared to controls. However, these pharmacological differences only emerge at 4-6 months of age--3 months after the appearance of obesity. High levels of immunoreactive endorphin in the pituitary are, therefore, more likely to be a consequence than a cause of obesity. Furthermore, numerous other neurologic abnormalities, which may or may not play a role in the obesity syndrome, are evident in ob/ob mice. Compared to controls, ob/ob total brain, hypothalamus, and pituitary weights are 11%, 16%, and 23% less, respectively. Levels of immunoreactive Leu5-enkephalin in pars nervous are also 200% higher in ob/ob mice; this increase is apparent at 1-6 months of age and is highly correlated with changes in body weight.


Release of catecholamines from perfused cat adrenal gland by veratridine. Experiments were undertaken to verify the existence of fast sodium channels in the adrenal chromaffin cell membrane and to assess their contribution to the physiological release of catecholamines. We have used veratridine to activate sodium channels. Veratridine causes secretion of catecholamines from perfused cat adrenal gland. Secretory response to veratridine is calcium dependent and abolished by tetrodotoxin. Secretion of catecholamines by acetylcholine is only partially blocked by tetrodotoxin. It is concluded that the adrenal chromaffin cell membrane contains fast sodium channels directly comparable to those of impulse-propagating neurons, but they do not appear to be essential in the secretory response to acetylcholine or splanchnic nerve stimulation.


Coevolution in structured demes.

Coevolution in structured demes. A simple model of coevolution in a subdivided population is considered. It is shown that, when there are frequency- and density-dependent interactions in each site, the sampling variation in numbers in each local site can lead to selection both through the dispersal process and through indirect effects. The model predicts that coevolved relationships between species can result from various interactions other than direct forms of competition and predation.


Geometric and electronic structure of oxyhemocyanin: spectral and chemical correlations to met apo, half met, met, and dimer active sites. The chemical and spectral properties of a series of hemocyanin derivatives were systematically compared to provide insight into the geometric and electronic structure of the oxyhemocyanin active site. The binuclear copper site is characterized as two tetragonal Cu(II) atoms bridged by both an endogenous protein ligand and the exogenous ligand (i.e., peroxide), with the lack of an electron paramagnetic resonance signal being the result of antiferromagnetic exchange via the endogenous bridge. A transition dipole-vector coupling model is used to assign the unique absorption spectral properties of oxyhemocyanin: the bands at 570 and 486 nm are assigned as components of the peroxide pi v* to copper dx2-y2 charge transfer. The 345-nm band is one component of the pi sigma* leads to dx2-y2 charge transfer. The model also predicts an end-to-end bridging geometry for the peroxide in oxyhemocyanin.


In situ phosphorylation of the alpha subunit of eukaryotic initiation factor 2 in reticulocyte lysates inhibited by heme deficiency, double-stranded RNA, oxidized glutathione, or the heme-regulated protein kinase. Protein synthesis initiation in reticulocyte lysates is inhibited by heme deficiency, low levels of double-stranded RNA (dsRNA), oxidized glutathione (GSSG), or the purified kinase (HRI) that acts on the alpha polypeptide of eukaryotic initiation factor 2 (eIF-2alpha). The phosphoprotein profiles produced in lysates in response to these various conditions have been monitored directly in lysates after labeling for brief periods with pulses of [gamma-(32)P]ATP. The [(32)P]phosphoprotein profiles were analyzed by electrophoresis in sodium dodecyl sulfate/polyacrylamide slab gels under conditions in which the HRI and eIF-2alpha polypeptides were clearly distinguished. All four modes of inhibition produced a rapid phosphorylation of eIF-2alpha compared to control lysates, which displayed little or no phosphorylation of eIF-2alpha. In heme-deficient lysates, phosphorylation of eIF-2alpha occurred rapidly both before and after the shut-off of protein synthesis; the delayed addition of hemin to these lysates resulted in a decrease in the phosphorylation of eIF-2alpha and the subsequent restoration of protein synthesis. These data suggest that rapid turnover of phosphate occurs at the site(s) of eIF-2alpha phosphorylation. In lysates inhibited by heme deficiency, GSSG, or added HRI, the phosphorylation of eIF-2alpha was accompanied by the rapid in situ phosphorylation of HRI. The inhibition of initiation induced by dsRNA was accompanied by the phosphorylation of eIF-2alpha and a 67,000-dalton polypeptide but not HRI. These observations in situ indicate that (i) the phosphorylation of eIF-2alpha is the critical event in these inhibitions of protein chain initiation, and (ii) the phosphorylation of HRI is associated with its activation in heme deficiency.


Albumin microspheres as carrier of an inhibitor of leukocyte elastase: potential therapeutic agent for emphysema. Methods are described for the covalent attachment of succinoyl-Ala-Ala-Pro-ValCH2Cl, an active site-directed inhibitor of human leukocyte elastase (EC 3.4.21.11), to microspheres of human albumin. The insertion of side arms of various lengths revealed that maximum inhibition of this enzyme was obtained when the spacer arm was at least 24.3 A in length. Approximately 30 molecules of the inhibitor could be attached to each molecule of albumin. Such derivatized microspheres were capable of inhibiting approximately one mole of elastase per mole of albumin, which is comparable to the inhibitory activity of alpha 1-antitrypsin. Experiments in vivo in which rats were injected intravenously with radiolabeled microspheres to which the inhibitor had been attached showed a rapid and exclusive uptake by the lungs. About 40--50% of the injected microspheres subsequently remained in the lungs with a half-life of approximately 17 days. These derivatized microspheres thus appear to offer promise as a therapeutic agent for emphysema.


Reaction of nucleosome DNA with dimethyl sulfate. We have measured the effect of the histones in the nucleosome core particle on methylation of purines in nucleosome DNA by dimethyl sulfate. By using 32P terminally labeled nucleosome cores, we have examined the pattern of strand cleavage at methylated sites in the nucleosome DNA and compared it to the pattern observed in histone-free DNA. We are unable to detect any significant difference between the reactivity of N7 of guanines in nucleosome DNA and of that in naked DNA, with the exception of a single site of enhanced reactivity at approximately nucleotide 62 from the 5' end of the nucleosome. Contrary to our expectation, there is no detectable periodic modulation of reactivity corresponding to the twist of the DNA on the nucleosome surface. We are able to place a low upper limit on the extent to which the histones of the nucleosome can protect N7 of guanine in the large groove. With somewhat less precision, we also conclude that the N3 of adenine in the small groove is largely unprotected. These results indicate that in nucleosome DNA the bases are nearly as accessible to solvent as they are in DNA free of protein.


Further studies on the mode of action of the heme-controlled translational inhibitor: stimulating protein acts at level of binary complex formation. Previous work has shown that (i) at physiological concentrations of eukaryotic initiation factor 2 (eIF-2), formation of the ternary complex eIF-2-GTP-Met-tRNAi, which precedes the assembly of a 40S initiation complex, requires the presence of eIF-2 stimulating protein (ESP) and (ii) the interaction of eIF-2 with ESP is blocked by the translational inhibitor which, in reticulocyte lysates, is activated in the absence of hemin. Present evidence indicates that formation of the ternary complex is preceded by formation of the binary complex eIF-2-GTP and that ESP acts at the level of binary complex formation.


Methods for identifying proteins by using partial sequences. Methods for the identification of a protein segment by using the information from partial sequence analyses are described. If the protein sequence is known, a segment can usually be identified with confidence through comparison with the data file of all known sequences when the identity and position of only seven amino acid residues (not necessarily contiguous) are known. Partial sequences are obtained from extremely sensitive microsequencing procedures. Tissue is incubated with amino acids, one or more of which are distinctively radiolabeled. Proteins of interest are isolated and a sequenator experiment performed to locate the positions of radioactivity in an NH(2)-terminal segment of approximately 30 residues. We derive and investigate an equation for the probability of finding a unique match to any pattern of radioactivity. From this we suggest a new strategy. In one incubation, several amino acids are labeled with each kind of isotope. The most information is contained in patterns in which approximately equal numbers of positions are occupied by residues distinguished by different labels (including no label). The amino acid composition of the segment will typically not be known in advance. Labeling residues expected to occupy 36% of the positions suffices for a 98% chance of success in uniquely characterizing any human segment. Such a strategy will permit the identification of most proteins from a single tissue incubation. The mathematical discussion is general and applies to any segment from a sequence and to sequences obtained by any method. Improved identification procedures should expedite the accumulation of information on the expression and function of proteins.


Mechanisms and prevention of trifluoroacetylation in solid-phase peptide synthesis. A novel mechanism for trifluoroacetylation in solid-phase peptide synthesis, independent of the coupling step, has been elucidated. It involves the presence of trifluoroacetoxymethyl groups on the resin support, which react with resin-bound amines by an intersite nucleophilic reaction. The trifluoroacetoxymethyl groups are generated from preexisting hydroxymethyl sites during treatment with trifluoroacetic acid in dichloromethane or by acidolysis of the benzyl ester bond between the peptide and the resin. The transfer of trifluoroacetyl from hydroxyl to amine occurs during the subsequent neutralization with tertiary amine. The mechanism was first elucidated by model studies with aminomethyl-resins. Then the expected transfer of trifluoroacetyl groups from trifluoroacetoxymethyl-resin to the alpha-amino group of N(epsilon)-benzyloxycarbonyllysine benzyl ester in solution was demonstrated; k(2), 6 x 10(-4) M(-1). Lysine-resins were used to examine the extent of trifluoroacetylation under the conditions of solid-phase peptide synthesis. After a series of acid/base cycles simulating synthetic conditions but without coupling, the poorly nucleophilic alpha-amino group was approximately 1-2% trifluoroacetylated per cycle when attached to resins already containing hydroxymethyl groups. Standard benzyl ester resins without preexisting hydroxymethyl groups gave comparable levels of trifluoroacetylation after the first few synthetic cycles because of gradual acid cleavage of the ester and accumulation of trifluoroacetoxymethyl sites. Peptide chain termination resulting from trifluoroacetylation by this mechanism could be prevented (<0.02% per cycle) by the use of the aminoacyl-4-(oxymethyl)-phenylacetamidomethyl-resin support, which can be synthesized free from extraneous functionalities and which is stable to trifluoroacetic acid under the conditions of solid-phase peptide synthesis.


Activation of phenotypic expression of human globin genes from nonerythroid cells by chromosome-dependent transfer to tetraploid mouse erythroleukemia cells. Chromosome-dependent gene transfer mediated by cell fusion was used to show that it is possible to activate phenotypic expression of human alpha globin genes derived from nonerythroid cells. Hybrid cells containing the human alpha globin structural genes were derived by fusion of populations of adult human peripheral blood mononuclear cells (devoid of identifiable erythroid cells) with adenine phosphoribosyl-transferase-deficient mouse erythroleukemia cells that contained close to a tetraploid complement of mouse chromosomes. The hybrid cells retained a near tetraploid complement of mouse chromosomes but had lost 80% of the chromosomes of the human parent cell. All of these hybrid cells and their subclones, which contained human chromosome 16, exhibited synthesis of human alpha globin chains. Human alpha globin mRNA was also demonstrated to be present in one of these hybrid cells by RNA.cDNA molecular hybridization analysis. We conclude that the mechanism responsible for restricting expression of the human globin genes in nonerthroid cells is not irreversible, at least for those globin structural genes that are actively transcribed in erythroid cells during adult life. Moreover, some genetic factor or process in the tetraploid mouse erythroleukemia cell is, under the conditions of our experiments, capable of reactivating phenotypic expression (production of globin chains) of human globin genes derived from nonerythroid hematopoietic cells after chromosome-dependent gene transfer.


Histone 1 is proximal to histone 2A and to A24. Water-soluble carbodiimide crosslinks histones 1A and 1B to histone 2A and to semi-histone A24 in chromatin from mouse cells. The identities of the histone species present in the crosslinked dimers were determined by fingerprinting. The molar ratio of H1--A24 to H2A is the same as the molar ratio of A24 to H2A in these cells. The H1-H2A crosslinks form equally well in whole nuclei, lysed nuclei, and H1-containing mononucleosomes isolated from a sucrose gradient. These results suggest that there exist major H1 interactions within the nucleosome.


Organization of a hybrid between phage f1 and plasmid pSC101. We have characterized the 200-nucleotide-long insertion found in f1 after segregation of a chimeric phage containing the genomes of f1 and pSC101 [Ohsumi, M., Vovis, G.F. & Zinder, N.D. (1978) Virology 89, 438--449]. The insertion in this novel f1 species, called f1', is derived from pSC101 and has the potential to form an extended base-paired secondary structure, as determined by nucleotide sequence analysis. A five-nucleotide direct repeat, derived from f1 sequences, is present in f1'. The 200 additional nucleotides that are inserted into the DNA sequence coding for the carboxy terminus of f1 gene IV protein have generated a novel carboxy terminus for the f1' gene IV protein. In vitro transcription--translation studies demonstrate that a read-through protein can be expressed, as predicted from the f1' nucleotide sequence results. This 200-nucleotide-long sequence appears to be a transposable element found within pSC101 and is similar in sequence to the inverted repeat found in Tn3. Restriction enzyme analysis of the chimeric phage DNA, coupled with the nucleotide sequencing results, allows us to predict a structure for the genomic organization of this chimera.


Effect of Cibacron blue F3GA on oligonucleotide binding site of estradiol--receptor complexes of mouse uterine cytosol. The binding of estradiol--receptor complexes of mouse uterine cytosol to oligodeoxynucleotide celluloses is inhibited by the sulfonated polyaromatic dye Cibacron blue F3GA. The dye does not have any effect on the estradiol binding site. Additon of the dye to preformed estradiol--receptor--oligo(dT)-cellulose complex results in the release of estradiol--receptor. The inhibition of binding is competitive with respect to oligo(dT)- or oligo(dA)-Celluloses, suggesting that the effect of the dye is directly on the polynucleotide binding site of the receptor. The observed inhibitory effect of Cibacron blue is not a simple electrostatic effect of polyanions because heparin and polyglutamic acid are much less effective. The selective inhibition by Cibacron blue suggests that the polynucleotide binding domain of estradiol--receptor possesses a special "supersecondary" structure [Stellwagen, E. (1977) Acc. Chem. Res. 10, 92--98].


Cellular retinol-binding protein allows specific interaction of retinol with the nucleus in vitro. Purified cellular retinol-binding protein (CRBP), a potential mediator of vitamin A action, was found to enable retinol to bind in a specific manner to isolated nuclei from livers of vitamin A deficient rats. Binding was followed after complexing [3H]retinol with CRBP. The binding was specific, saturable, and temperature dependent. CRBP charged with unlabeled retinol or CRBP without retinol diminished binding of radioactivity whereas free retinol did not. No specific binding sites could be detected for free retinol. Purified cellular retinoic acid binding protein (CRABI) complexed with retinoic acid did not diminish the amount of retinol bound to nuclei. Approximately 3 x 10(5) specific binding sites per nucleus could be detected. Fewer binding sites were found in nuclei isolated from livers of control (chow-fed) rats and also from livers of vitamin A-deficient rats 2 hr after refeeding with retinylacetate.


Distinct single-copy sequence sets in sea urchin nuclear RNAs. The purpose of this study was to determine whether nuclear RNAs (nRNA) of sea urchin embryos and adult tissues contain identical or partially distinct sets of single-copy sequence transcripts. A DNA tracer was prepared consisting mainly of sequences absent from gastrula nRNA; 3.6% of this tracer reacted with adult intestine nRNA but not with gastrula nRNA. The existence of a differentially transcribed DNA fraction was verified by its partial purification and rehybridization to intestine and gastrula nRNAs. About one-third of the genomic single-copy sequence is represented in both nRNAs, or about 2 x 10(8) nucleotides. The differentially transcribed portion of the single-copy genome identified in this work includes about 3.5 x 10(7) nucleotides.


Inhibition of human neuroblastoma DNA polymerase activities by plant lectins and toxins. The effects of concanavalin A and ricin (RCAII, Mr 65,000) on [3H]thymidine incorporation into human neuroblastoma IMR-32 DNA showed reduction of total DNA synthesis to 50% and 70% of control, respectively. Two DNA polymerase (DNA nucleotidyltransferase, EC 2.7.7.7.) activities (alpha and beta) involved in the biosynthesis in vitro of DNA were separated by sucrose density gradient centrifugation from IMR-32 cell homogenate. The DNA polymerase alpha activity was also purified by selective precipitation with polyethylene glycol (Mr 6000) followed by agarose-concanavalin A column chromatography. The activities of both DNA polymerases were examined at various concentrations of mutagenic and nonmutagenic plant agglutinins and the toxin ricin. Concanavalin A and ricin specifically inhibited DNA polymerase alpha activity (activity reduced to 19% and 10%, respectively), whereas DNA polymerase beta activity was inhibited (reduced to 16%) by red kidney bean agglutinin (PHA-P).


Structure-independent nucleotide sequence analysis. Substitution of inosine for granosine in the nucleic acid fragments synthesized for the sequencing of RNA effectively prevents the formation of secondary structures during electrophoretic analysis. Consequently, the mobility of each fragment in the sequencing gel is a strict function of its molecular weight. Inosine substitution should markedly improve the resolution that can be obtained in the sequencing of DNA as well as RNA.


Steady-state coupling of four membrane systems in mitochondrial oxidative phosphorylation. According to Alexandre, Reynafarje, and Lehninger, four different membrane systems are involved, with definite stoichiometry, in the mitochondrial synthesis of ATP by electron transport, via proton transport. We adopt this model and pursue some of its thermodynamic consequences. At steady state, each of the four systems must have the same flux J through the membrane and the overall thermodynamic force X for oxidative phosphorylation is the sum of the four separate forces. From these properties, using an empirical linear flux-force relation for each system, it is easy to obtain J as a function of X. In turn, X depends on the inside [NAD+]/[NADH] and the outside [ATP]/[ADP][Pi] quotients (and on the pH inside). Thus, J is related to these quotients. The relationship we derive is similar to that described by Erecińska and Wilson, as deduced from a quite different model of oxidative phosphorylation. Proton transport is involved explicitly in three of the four systems of the present model. However, because of the steady-state stoichiometric coupling of the four systems, proton transport does not appear in the overall reaction. On the other hand, Erecińska and Wilson use, in their model, a direct connection between electron transport and ATP synthesis. The present paper demonstrates that J can be related to the quotients mentioned above without this direct connection.


Thermotropic behavior of some fluorodimyristoylphosphatidylcholines. The thermotropic behavior of three fluorinated phospholipids has been studied by means of high-sensitivity differential scanning calorimetry. The three lipids are 1,2-di(X,X-difluoromyristovyl)phosphatidylcholine with X = 4,8, or 12. The compound with X = 4 has a higher transition temperature than dimyristoylphosphatidylcholine whereas the other two have lower values. All three lipids have transition enthalpies approximately twice that of the unsubstituted lipid. It was found the substituted lipids form strikingly nonideal mixtures with the parent compound. In view of these observations it is evident that results obtained using these or related compounds as probes of bilayer or membrane behavior must be interpreted with due caution. Some of the properties exhibited by geminal fluorine atoms in fluorinated phospholipids appear to be similar to those of cis double bonds in lipids.


The ovalbumin gene: cloning and molecular organization of the entire natural gene. We report the analyses of recently cloned restriction fragments of the natural ovalbumin gene that overlap in part with previously cloned DNA fragments but extend further into the flanking sequences of the gene. These clones now permit us to identify the DNA sequence that codes for the 5' end of ovalbumin mRNA. Based on these and previous results, the molecular organization of the entire ovalbumin gene was established. The entire gene is composed of eight structural DNA sequences separated by seven intervening sequences that are not present in the mature mRNA. In addition, an ovalbumin gene clone has been obtained from a chicken gene library. Analysis of DNA isolated from this particular clone by molecular hybridization and electron microscopic mapping revealed that it contains the entire ovalbumin gene a single segment of DNA and its structure was consistent with that predicted from our physical map constructed from individually cloned fragments of the gene.


Cell-free translation of the mRNAs for the heavy and light chains of HLA-A and HLA-B antigens. Cell-free translation of poly(A)-containing RNA from the JY lymphoblastoid cell line followed by immunoprecipitation has indicated the presence of mRNAs for both the heavy and light chains of HLA-A and HLA-B antigens in these preparations. Both chains are synthesized with an NH2-terminal extension, approximately 20 residues in length for the light chain, and 20 or 24 residues for the heavy chains. The precursors can be processed by dog pancreatic microsomes to products similar to those obtained in vivo. Immunoprecipitation of the cell-free products has been employed as an assay for partial purification of the mRNAs. Investigation of the Daudi cell line, which cannot synthesize the small subunit, beta 2-microglobin, has indicated that the heavy chains of HLA-A and HLA-B antigens are synthesized intracellularly in vivo and can also be translated from their cognate RNAs in vitro. The implications of these findings for biosynthesis of membrane proteins in general and multimeric membrane proteins in particular, as well as the role of beta 2-microglobulin in expression of HLA-A and HLA-B antigens, are discussed.


Macrophage response to concanavalin A: effect of surface crosslinking on the electrophoretic mobility distribution. Electrophoretic light scattering (laser Doppler electrophoresis) has been employed to study the effects of concanavalin A (Con A) and succinyl-Con A on the electrophoretic mobility distribution of resident guinea pig peritoneal macrophages. Con A, a tetrameric lectin, decreases slightly the mean mobility and increases by approximately 3-fold the width of the electrophoretic mobility distribution of resident macrophages. This effect can be abolished by alpha-methyl-D-mannoside, a hapten sugar of Con A. These effects were present in both low (0.010 M) and high (physiological, 0.15 M) ionic-strength media. Since lower ionic strengths correspond to a larger Debye screening distance, these data suggest that the alterations in the electrophoretic mobility distribution are not restricted to the outer portion of the glycocalyx. Succinyl-Con A, a dimeric derivative, was found to have no effect on the mobility distribution. However, the mean mobility decreased and the width increased over 4-fold when succinyl-Con A-treated macrophages were exposed to anti-Con A. These observations indicate that cross-linking of Con A receptors is an important process in the electrokinetic alterations of the macrophage surface. These results may have important consequences for the elucidation of the details of the endocytic mechanism.


The rate of access to the organic ligand-binding region of serum albumin is entropy controlled. The technique of real time dielectric relaxation measurement coupled with a conventional stopped-flow device has made it possible to measure the rates of association and dissociation of the complex of human serum albumin with its most prevalent ligands, the long-chain fatty acids. This association was previously shown to proceed in two steps: a fast, probably diffusion-controlled, nonspecific association, followed by a slower (approximately 3 sec--1) rearrangement of the intermediate protein--ligand configuration, whose kinetics is first order. By use of the Arrhenius relation and standard theory of rate processes it is determined that there is virtually no activation enthalpy in the forward binding reaction and that the rate of access to the interior hydrophobic binding region of serum albumin is controlled by a negative entropy of activation, reflecting a high degree of ordering in the transition state. A complete thermodynamic and kinetic profile of the association reaction is given.


Diamagnetic anisotropy of the peptide group. A simple theory of the diamagnetic anisotropy of noncyclic planar groups of atoms with resonance structures (mobile electrons)leads to the value --5.36 x 10(-6) cm-g-sec electromagnetic units for the molar diamagnetic anisotropy of the peptide group.


Complexity transmission during replication. The transmission of complexity during DNA replication has been investigated to clarify the significance of this molecular property in a deterministic process. Complexity was equated with the amount of randomness within an ordered molecular structure and measured by the entropy of a posteriori probabilities for discrete (monomer sequences, atomic bonds) and continuous (torsion angle sequences) structural parameters in polynucleotides, proteins, and ligand molecules. A theoretical analysis revealed that sequence complexity decreases during transmission from DNA to protein. It was also found that sequence complexity limits the attainable complexity in the folding of a polypeptide chain and that a protein cannot interact with a ligand moiety of higher complexity. The analysis indicated, furthermore, that in any deterministic molecular process a cause possesses more complexity than its effect. This outcome broadly complies with Curie's symmetry principle. Results from an analysis of an extensive set of experimental data are presented; they corroborate these findings. It is suggested, therefore, that complexity governs the direction of order-order molecular transformations. Two biological implications are (i) replication of DNA in a stepwise, repetitive manner by a polymerase appears to be a necessary consequence of structural constraints imposed by complexity, and (ii) during evolution, increases in complexity had to involve a nondeterministic mechanism. This latter requirement apparently applied also to development of the first replicating system on earth.


Upper surfaces of epithelial sheets and of fluid lipid films are nonadhesive for platelets. Platelet-rich rabbit plasma was incubated with surfaces of two different types--sheets of cultured mouse kidney epithelium and films of different lipids. The upper surface of epithelial sheets was found to be nonadhesive for platelets; in the same cultures, the platelets attached easily to the glass surface not covered by epithelial cells. Platelets did not attach to the surface of lipids (egg lecthin and dioleoyllecithin) that were in the liquid-crystalline state at 23 degrees C. In contrast, the surface of the films made of lipids (dipalmitoyl- and distearoyllecithin) that were in the crystalline state at 23 degrees C was adhesive for platelets. It had been found previously that the surfaces of epithelial sheets and of liquid lipid films are nonadhesive for fibroblasts. Possible mechanisms responsible for the nonadhesiveness of these surfaces are discussed. It is stressed that the factors responsible for nonadhesiveness of epithelial surfaces may be similar to those responsible for nonadhesiveness of the luminal surface of endothelium in blood vessels.


Appearance in confluent vascular endothelial cell monolayers of a specific cell surface protein (CSP-60) not detected in actively growing endothelial cells or in cell types growing in multiple layers. The formation of a highly organized vascular and corneal endothelial cell monolayer is associated with the appearance of a 60,000-dalton cell surface protein (CSP-60) (30,000 daltons after reduction with dithiothreitol) which is not detectable in rapidly growing endothelial cells and in subconfluent cultures that do not yet exhibit the strict morphology of a confluent monolayer. It is also absent from vascular smooth muscle cells and from endothelial cultures that are maintained in the absence of fibroblast growth factor and grow on top of each other at confluence. After disorganization of cells in a confluent endothelial monolayer by urea, EDTA, or trypsin, CPS-60 is no longer exposed on the cell surface, but it reappears as soon as the cells readopt their characteristic two-dimensional configuration. This reorganization can be achieved in the presence of cycloheximide and despite removal of fibronectin by urea, EDTA, or trypsin. Maximal amounts of fibronectin and no CSP-60 are detected in subconfluent, but not yet organized, endothelial cultures or in endothelial cells that no longer form a monolayer of nonoverlapping cells at confluence. Likewise, cultures of vascular smooth muscle cells contain fibronectin but no CSP-60. These results suggest that CSP-60, rather than fibronectin, could be involved in the adoption of a monolayer configuration by confluent endothelial cells.


Functional roles of DNA polymerases beta and gamma. The physiological functions of DNA polymerases (deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase, EC 2.7.7.7) beta and gamma were investigated by using neuronal nuclei and synaptosomes isolated from rat brain. UV irradiation of neuronal nuclei from 60-day-old rats resulted in a 7- to 10-fold stimulation of DNA repair synthesis attributable to DNA polymerase beta which, at this developmental stage, is virtually the only DNA polymerase present in the nuclei. No repair synthesis could be elicited by treating the nuclei with N-methyl-N-nitrosourea, but this way probably due to the inability of brain tissues to excise alkylated bases from DNA. The role of DNA polymerase gamma was studied in synaptosomes by using a system mimicking in vivo mitochondrial DNA synthesis. By showing that, under these conditions, DNA replication occurs in mitochondria, and exploiting the fact that DNA polymerase gama is the only DNA polymerase present in mitochondria, evidence was obtained for a role of DNA polymerase gamma in mitochondrial DNA replication. Based on these results and on the wealth of literature on DNA polymerase alpha, we conclude that DNA polymerase alpha is mainly responsible for DNA replication in nuclei, DNA polymerase beta is involved in nuclear DNA repair, and DNA polymerase gamma is the mitochondrial replicating enzyme. However, minor roles for DNA polymerase alpha in DNA repair or for DNA polymerase beta in DNA replication cannot be excluded.


Villin: the major microfilament-associated protein of the intestinal microvillus. The major protein associated with actin in the microfilament core of intestinal microvilli has been purified. This protein, for which we propose the name villin, has a polypeptide molecular weight of approximately 95,000. Two arguments suggest that villin may be the microvillus crossfilament protein that links the microfilament core laterally down its length to the cytoplasmic side of the plasma membrane. First, electron microscopy shows that crossfilaments stay attached to isolated membrane-free microvillus cores. Calculation of the expected abundance of the crossfilament protein shows that only villin is present in sufficient quantity to account for these structures. Second, decoration of microvillus cores by antibodies to either actin or villin, followed by ferritin-labeled second antibody in a sandwich procedure, results in specific labeling of the cores in both cases. The antivillin decoration, however, gives rise to a greater increase in diameter, in agreement with a model in which villin projects from the F-actin microfilament core. Villin is distinct from alpha-actinin, a protein suggested to be involved in membrane anchorage of microfilaments in nonmuscle cells. The two proteins differ in molecular weight. Specific antibodies against villin and alpha-actinin show no immunological crossreactivity. Immunofluorescence microscopy reveals that villin is located in the microvilli of the brush border whereas alpha-actinin is absent from the microvilli but is found in the terminal web. In addition, villin is not found in microfilament bundles of tissue culture cells, which are rich in alpha-actinin. Thus, villin and alpha-actinin appear to be immunologically and functionally different proteins.


Direct demonstration of binding of a lysosomal enzyme, alpha-L-iduronidase, to receptors on cultured fibroblasts. Receptor-binding of "high-uptake" forms of lysosomal enzymes to human diploid skin fibroblasts had been predicted from the Michaelis--Menten kinetics of uptake of these enzymes [e.g., Sando, G.N. & Neufeld, E.F. (1977) Cell 12, 619--627]. We have now demonstrated such binding directly by using a sensitive assay for the bound enzyme. Cells deficient in alpha-L-iduronidase were detached from plastic dishes by mild trypsinization, allowed to recover, and used in suspension. They were incubated with urinary alpha-L-iduronidase at 0 degrees C for 90 minutes and then washed by centrifugation through concentrated bovine serum albumin; the activity of the cell-associated enzyme was measured with 4-methylumbelliferyl alpha-L-iduronide as substrate. A Scatchard analysis showed 14,000 binding sites per cell and a Kd of 1 x 10(-9) M for high-uptake alpha-L-iduronidase; binding of the low-uptake form was barely detectable. Mannose 6-phosphate, a known competitive inhibitor of uptake, inhibited the binding competitively, with Ki = 1 x 10(-4) M. Unexpectedly, mannose 6-phosphate greatly accelerated the dissociation of bound enzyme. During uptake of alpha-L-iduronidase at 35 degrees C, the receptors were regenerated every few minutes, even in the absence of protein synthesis.


Syndeins: the spectrin-binding protein(s) of the human erythrocyte membrane. Two-dimensional tryptic and chymotryptic analyses of all the major bands in a sodium dodecyl sulfate/polyacrylamide gel of the human erythrocyte membrane show that each band has a characteristic map. However, band 2.1 (nomenclature of T. L. Steck) and several polypeptides below this band exhibit similar tryptic and chymotryptic peptide maps and thus appear to be a family of closely related proteins or degradation products. Furthermore, they all contain a subset of peptides that are accounted for by the peptides from two known spectrin-binding fragments. We show that both fragments derive from 2.1-related proteins and conclude that band 2.1 and its related proteins, which we name "syndeins", bind spectrin and connect it to the erythrocyte membrane.


Actin polymerization induced by a motility-related high-affinity cytochalasin binding complex from human erythrocyte membrane. A high molecular weight complex (sedimentation coefficient approximately 27 S) containing high-affinity binding site(s) for [(3)H]dihydrocytochalasin B has been isolated from a low ionic strength extract of human erythrocyte membranes by sucrose density gradient centrifugation. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis showed that actin, spectrin, and other minor components, including two polypeptides with the electrophoretic mobility of band 4.1, were present in the complex-containing fraction. Addition of this complex to a solution of muscle monomeric actin (G-actin) in a low ionic strength medium resulted in a rapid increase in viscosity to a level comparable to that of a solution of filamentous actin (F-actin). Electron microscopy showed that the viscosity increase reflected actin filament formation. The rate of induced actin polymerization was dependent on the amount of complex added to the G-actin; in less than 1 hr, less than 1 mug of protein from the complex-containing fraction induced the conversion of 0.4 mg of G-actin to the "F" from. Binding studies indicated that, upon polymerization of the actin, the cytochalasin binding complex became associated with the actin filaments. Low concentrations of cytochalasins D and E and dihydrocytochalasin B inhibited actin polymerization induced by the complex; the relative potencies of the drugs in inhibiting this process corresponded to their relative affinities for the complex, as well as their relative potencies in affecting cell motility. These results suggest that the cytochalasin binding complex functions as a regulatory site for cell motility by controlling formation and membrane attachment of actin-containing microfilaments in the cell.


Sodium-induced calcium release from mitochondria in brown adipose tissue. Coupled mitochondria of brown adipose tissue can accumulate Ca2+ if a substrate is present. The Ca2+ is released by addition of 20 mM Na+, but not by addition of K+ or choline +. Energy-dissipating Na+-induced Ca2+ cycling occurs maximally with 20 mM Na+ and 10 microM Ca2+. In brown adipocytes, the Ca2+ ionophore A23187 and the Na+ ionophore monensin increase respiration if substrate is added, and incubation in a low-Na+ buffer decreases norepinephrine-induced respiration. Thus Na+-induced Ca2+ release takes place in brown adipose tissue; released Ca2+ could have a regulatory or thermogenic role or both.


Partial deletion of beta-globin gene DNA in certain patients with beta 0-thalassemia. We have used restriction endonuclease mapping of cell DNA to investigate the structure of the beta-globin gene in beta-thalassemias. Among 17 individuals with beta +- and beta 0-thalassemia, we observed three patients of Indian origin with beta 0-thalassemia whose DNA revealed a consistent mapping abnormality. In one beta allele in each diploid cell, 0.6 kilobase of DNA was deleted from  beta-specific Pst I and Bgl II restriction fragments. This deletion involved 3' beta-globin gene sequences and eliminated the EcoRI site normally present at codons 121/122, but it did not extend to the BamHI site at codons 98--100 on the 5' side of the 0.90-kilobase intervening sequence normally present in beta-globin genes. Partial beta-globin gene deletion appears, therefore, to be a primary molecular defect seen in certain patients with beta 0-thalassemia.


Pregnant mice are not primed but can be primed to fetal alloantigens. In this report we present evidence gathered from various stages of murine pregnancy that indicates that pregnant females have no demonstrable immune effector function directed against their semiallogeneic fetuses. Specifically, by using in vitro assays we found that cytotoxic lymphocytes were not present in pregnant mice, and pregnant mice challenged with radiolabeled paternal strain leukemia cells showed no evidence of immune elimination. Nonetheless, immunity measurable both in vitro and in vivo was readily induced by priming with the paternal strain cells. No harm to the fetus was observed in primed mothers. These results cast doubt on the relevance of mechanisms proposed that involve systemic active suppression during pregnancy. The results are compatible with the hypothesis that the placenta acts as a barrier, preventing fetal cells from priming the mother.


Serum iron levels and response to hepatitis B virus. Response to hepatitis B virus (HBV) infection [HBV surface antigen (HBsAg) and antibody to HBsAg (anti-HBs)], serum iron, total iron-binding capacity, hematological status (erythrocytes, Hb, and hematocrit), and evidence of liver damage (serum glutamic pyruvic transaminase; aspartate aminotransferase, L-aspartate:2-oxoglutarate aminotransferase, EC 2.6.1.1) were determined for 201 patients on chronic renal dialysis. Four factors-serum iron level, transminase level, sex, and HBV response [i.e., infected-HBsAg(+) (HBsAg positive), anti-HBs(+) (anti-HBs positive), or no response]-were analyzed simultaneously to test the hypothesis that serum iron is higher in those with HBsAg in their serum than in those without HBsAg, independent of the transaminase level. Four independent, statistically significant two-factor interactions were identified. (i) Serum iron is higher in those HBsAg(+). (ii) Serum iron is higher in those with increased transaminase. (iii) Transaminase is higher in those HBsAg(+). (iv) Males are more likely to be HbsAg(+) and females are more likely to be anti-HBs(+). Also, those who are HBsAg(+) have significantly higher percent iron saturation (serum iron/total iron-binding capacity). That is, the hypothesis was supported by the findings. Several additional biological hypotheses are suggested, including a possible role of increased iron levels in susceptibility and response to HBV infection and the possible relationship between higher iron levels and the likelihood of HBV infection progressing to primary hepatocellular carcinoma. In addition, further tests of the initial hypothesis in nonhospitalized populations with endemic HBV infection are proposed.


Release and distribution of [3H]norepinephrine in nonpigmented and pigmented rabbit iris. Unequal accumulation of drugs in pigmented and nonpigmented iris has been known for some time. However, at low concentrations of the sympathetic neurotransmitter norepinephrine, the accumulation is equal in both types of irides. The spontaneous and potassium-induced release of 3H was studied in isolated rabbit irides labeled with 0.1 micron [3H]norepinephrine ([3H]NE). The spontaneous outflow and the tissue 3H content were similar in both types of irides, but the potassium-induced release was significantly higher in nonpigmented iris. When irides were labeled with [3H]NE in the presence of 1 micron desmethylimipramine, the potassium-induced release was still significantly higher in nonpigmented than in pigmented iris, although the tissue 3H content was higher in the latter. Subcellular distribution of [3H]NE in both types of irides is presented. The data indicate that [3H]NE is distributed unequally in both types of irides in spite of equal tissue accumulation. The binding of NE by the pigment cell suggests an additional site of loss, or site of uptake, of the transmitter in melanin-containing tissues. The physiological significance of such an unequal release in relation to the evolutionary adaptation of the eye to light is discussed.


A sciatic nerve protein has a trophic effect on development and maintenance of skeletal muscle cells in culture. Citrate-soluble extracts of chicken sciatic nerve were fractionated biochemically and added to cultures of embryonic chicken skeletal muscle in order to identify the component that exerted trophic influences on the muscle. A protein fraction that expressed trophic activity was obtained by ion-exchange chromatography on DEAE-cellulose followed by gel filtration on Sephadex G-100 superfine. This fraction enhanced the rate and degree of morphological maturation and the level of protein synthesis in embryonic muscle cells. Muscle fibers treated with this fraction after maturation in culture survived for longer periods in vitro than did comparable controls. Characterization of the active protein fraction by sodium dodecyl sulfate/polyacrylamide gel electrophoresis revealed the presence of one major protein (molecular weight 84,000) and three minor proteins. Electrophoretic analysis of biologically inactive gel filtration fractions indicated that the three minor proteins were contaminants and that biological activity was associated with the protein of molecular weight 84,000. Analytical isoelectric focusing revealed that the active protein was acidic and focused as four species with isoelectric points (pI) of 5.74, 5.77, 5.92, and 6.15. Maximal incorporation of [14C]-leucine into muscle cell protein was elicited by 20 microgram of active protein per culture dish. These data suggest that an acidic protein having trophic influences upon muscle has been identified and partially purified.


[An extraosseous osteoblastic osteosarcoma in the breast mimicking a pulmonary coin lesion on the chest radiograph (author's transl)]. Case report of an osteoblastic osteosarcoma in the breast, which in addition to its unusual presentation caused some differential diagnostic problems initially because it mimicked a pulmonary coin lesion. One year later a metastasis was noted which had the same x-ray features as the primary tumor.


Liver abnormalities in penicillamine treated rheumatoid arthritis. Liver enzymes were followed in 99 patients treated with D-penicillamine for rheumatoid arthritis. In six abnormalities were found which consisted of elevated levels of lactic dehydrogenase. ALAT/ASAT, alkaline phosphatases or combinations of these. The changes were reversible on stopping the drug with one possible exception. No evidence of biliary cirrhosis, chronic active hepatitis or HBag-associated hepatitis was found. Liver biopsy was performed in 4 cases--one was taken 2 months after the treatment was discontinued, and was normal. One biopsy showed mild inflammatory changes, whereas in two histologic evidence of toxic liver necrosis was present. Liver damage should be included among possible complications of D-PA treatment.


The inhibitory effect of D-penicillamine on human lymphocyte cultures stimulated by phytohaemagglutinin, the antagonistic action of L-cysteine and synergistic inhibition by copper sulphate. The inhibitory effects of D-penicillamine (D-Pen) on lymphocyte activation by PHA are found to be dose-dependent, showing significant effects above a concentration of 50 microgram/ml. The cells show a diminished sensitivity to D-Pen added after 24 hours in culture, although the dose-response relationship is maintained. L-cysteine is usually found to have an insignificant effect on PHA stimulation, although enhancement and inhibition are also sometimes observed. The supplementation of culture medium with L-cysteine abolishes the inhibitory effect of D-Pen, whilst copper sulphate synergistically inhibits PHA-induced transformation. The balance between the availability of naturally occurring thiols, copper ions and administered D-Pen may be critical in determining the eventual effect of the drug on the lymphoid system. Lymphocyte suppressive soluble factors appear in the sera of patients with rheumatoid arthritis and are modified by treatment with D-Pen. Although our experiments suggest possible modes of action of the drug on the immune system as a therapeutic agent as well as a potent inducer of autoimmunity in its own right, further work is required to delineate its precise mechanism of action.


Penicillamine in rheumatoid arthritis: clinical pharmacology and biochemical properties. The current status of the clinical pharmacology of penicillamine was reviewed. Its indications in the therapy of RA were defined, and current principles of dosage were presented. The autoimmune side effects were discussed in the light of their possible implication with regard to a locus of action of the drug on the immunological system. A comparison was presented of the biochemical properties of penicillamine and 5-thiopyridoxine, another mercaptan compound which appears to demonstrate a penicillamine-like action in patients with RA. It was found that 5-thiopyridoxine did not possess copper chelating properties, it failed to form a mixed disulfide with cystine, it did not induce dermolathyrism in the weanling rat, and it was not a vitamin B6 antagonist. If both of these compounds do work by a common mechanism in RA, then the aforementioned biochemical properties of penicillamine must be presumed to be not relevant to its fundamental action in this disease.


Penicillamine and collagen metabolism.

Penicillamine and collagen metabolism. Penicillamine blocks the crosslinking of collagen in man and in all the animal species it has been tested in. At the standard dose in which it is used it binds to the aldehydes present in collagen and prevents them from forming crosslinks. It is more efficient in inhibiting the crosslinking of soft tissues, where the aldehydes are derived from lysine, and less effective in preventing crosslinking of bone, where the aldehydes involved are primarily those derived from hydroxylysine. At very high doses it can inhibit lysyloxidase activity by making Cu unavailable to the enzyme.


Spleen size and chromosome analysis as prognostic factors in chronic myelogenous leukemia. Of 72 patients with chronic myelogenous leukemia (CML), 37 were Ph1 positive, eight were negative, and 27 were not evaluated for the Ph1 chromosome. Of the 37 patients positive for Ph1, 31 had adequate evaluation. These 31 patients were reviewed for survival with reference to spleen size. Seventeen patients with spleens 3 cm or less below the left costal margin had a 50% survival of 75 months, with a mean survival of 53.9 months. Fourteen patients with spleens greater than 3 cm below the left costal margin had a 50% survival of 21 months, with a mean survival of 19 months (P = .0014). It is concluded that small splenic size in CML is associated with longer survival. It may be that this observation should be taken into consideration when considering splenectomy in the management of patients with CML.


Chronic granulocytic leukemia: long-term remission in a patient with familial sarcoidosis. A patient is described who has chronic granulocytic leukemia (CGL) and familial sarcoidosis, and who has been in complete remission for six years after a single course of busulfan therapy. Her bone marrow Philadelphia chromosome persists and the leukocyte alkaline phosphatase activity remains subnormal. Leukocyte alkaline phosphatase levels in nine white patients with sarcoidosis were all found to be low. Bone marrow chromosomes from two patients with active sarcoidosis showed no Philadelphia chromosome, but one of them, and our patient with CGL, exhibited a pericentric inversion of chromosome 9.


Protection of the intestine during radiation of the pelvic bone. We have given a short course of intensive radiotherapy to two patients with osteogenic sarcoma of the right iliac bone while protecting the intestine by displacement. With the follow-up period now being a year and a half and two years, there is no evidence of injury to the intestine. Insertion of the pack does not require major dissection, and its removal is not traumatic. Both patients had displacement of viscera on the right side, but there is no reason anatomically why this procedure cannot be applied for similar lesions on the left side.


Excretion of glucose-containing oligosaccharides in urines of orthopedic patients. Urinary neutral oligosaccharides of various connective tissue diseases were studied by gel-filtration through Sephadex G--10 after treatments with cetylpyridinium chloride (CPC), Dowex 50 (H+ form) and Dowex 1 (Cl- form), in succession. Increased excretion of urinary glucose-containing oligosaccharides, specifically glucosylgalactose was observed in most of the patients with chondrosarcoma, rheumatoid arthritis, Werner's syndrome, Rothmund Thomson syndrome and Morquio's disease. However, urinary excretion of neutral oligosaccharides in the patients with osteosarcoma and other tumorous conditions, and some systemic disorders in the connective tissues, examined in the present study, showed almost normal values. It is indicated, therefore, that the activity of glucosidase in insufficient for the glucose-containing oligosaccharides produced from the ground substance(s) in the former type connective tissue diseases.


[Fissure contents in rodent molars].

[Fissure contents in rodent molars]. The fissure contents in rodent molars were examined by means of a stereomicroscope and compared with those in caries-free human molars and premolars. As to colour, consistency and removableness, the fissure contents in the rodent molars were similar to those in human teeth. An orange substance as found in the fissures of human teeth was not detected in the fissures of rodent molars. This fact does not tell against the use of rodent molars as a study model for fissure contents.


[Electromyographic studies of the oral region before and after treatment with the function regulator]. Electromyographic measurements of the activity of the lips were performed on 101 children with dental malocclusions. It was stated that the intensity of the tetanic activities depended on the type of malocclusion. On mouth closing the lower lip was in general more active than the upper lip. Electromyographic examinations before and after treatment with the function regulator showed a considerable decrease in muscular activity compared to the initial values. These results permit to conclude that the function regulator as a training tool in the oral vestibule exerts a beneficial effect on incorrect positions and parafunctions of the muscles of the lips.


[Dental treatment of kindergarteners at Halle]. A 3-year longitudinal study on kindergarteners revealed an increase in the number of children having need for dental treatment, from 30.1% in the youngest age-group to 58.7% in the oldest age-group. In the framework of a program of general dental treatment, dental examinations were conducted by stomatologists of Halle on the oldest age-groups. The following results were obtained: children who had received complete dental treatment, 71%; children who had received partial dental treatment, 13%; and children who had received no dental treatment, 16%. As only group treatment will ensure optimal treatment results, whereby the attitude of the kindergarten teachers is of considerable importance, its inclusion in the educational programs of the kindergartens is advocated.


[Effect of subjective evaluation on the reproducibility of the gingivitis indices]. The gingival index, gingivitis score, sulcus bleeding index, PM(A) index, selective gingiva index, periodontal index and periodontal disease index were verified with regard to the effect of subjective evaluation on their reproducibility. The authors studied in particular the utility of these indices in case controls and in the detection of minimal changes in the inflammatory process. Due to the numerical classification, the evaluation of the extent and of the degree of severity of the gingivitis requires an subjective decision and, consequently, the choice of an index depends on the objective of the examination. For the purpose of comparison with biometrical parameters, the authors presented an index (selective gingiva index) elaborated by themselves.


[Diagnostic and therapy-assessing studies of gingival capillary function by photoplethysmography (preliminary report)]. In the sense of relation pathology, the local peripheral blood supply plays a decisive role also in periodontal diseases. Thus, the differentiation of local circulatory disturbances is of great importance in diagnosis and early detection as well as in therapeutical decisions. Photoplethysmography is well suited for this purpose, especially in evaluating the curves by determining the time of inclination and the plateau index (if necessary, with resort to the nitrangin test). The indicated method also permits to assess the success of therapeutical measures.


[Microbiological studies in maxillofacial inflammations]. 297 materials from 446 patients with pyogenic infections were microbiologically evaluated. It was stated that Staphylococcus aureus is still the main causative organism (in spite of a downward tendency) and that its penicillin resistance is again increasing. The other gram-positive cocci occurred less frequently as pure cultures and more often as mixed cultures. The gram-negative germs increased with the increase in sensitivity to chemotherapeutical agents. 159 materials remained sterile. It must be assumed that this is in part attributable to anaerobes so far not determined. These results permit to draw the following practice-relevant conclusions: The territorial resistogram determines the use of chemotherapeutical agents; and it seems advisable not to employ penicillin in outpatient practice. Where the use of an antibiotic is indicated, treatment with oxytetracycline or gentamycin should be started.


[Comparative materials science studies of Evicrol plastic filling material]. The water absorption of Evicrol, its hardness, flexural strength and abrasion resistance were measured and compared with the values found for Adaptic and those for Kallocryl A as a non-filled polymethyl methacrylate plastic. The flexural strength and the abrasion resistance of Evicrol amount to those of Adaptic. The hardness of Evicrol falls also short of that of Adaptic. On the contrary, Evicrol behaves quite differently under water. It was found that the amound of water absorbed by Evicrol is some 3-fold greater than that absorbed by Adaptic, and that saturation is reached considerably slower.


[Surface problems in preparing Evicrol fillings]. The polymerization of Evicrol, like that of other composites, is adversely affected by atmospheric oxygen. This negative effect may be avoided by the use of matrices. On filling class V cavities, the utilization of well adaptable cellophane, aluminium and tin foils is justified. To obtain satisfactory results in final polishing, it is necessary to employ polishing disks suited for polishing the anorganic fillers contained in this restorative plastic.


[Occurrence of vascular canals in alveolar bone]. The authors present the results from the investigation of 414 full-mouth X-rays from periodontally diseased individuals for the occurrence of visible bone canals. The frequency, the form the most frequent location and the age-distribution of the vascular canals as well as the possibility of classifying them according to type and severity of the periodontal disease are discussed. Follow-up examinations in 18 patients, on an average 15 years after the first examination, permit the conclusion that the bone canals cannot be influenced by periodontal treatment, and that they are not associated with the periodontal disease.


[Interferences with the development of the jaws and teeth in mixed dentition]. On the basis of results from animal experiments, the authors emphasize a series of factors interfering considerably with tooth eruption. It is dealt with tooth eruption disorders and dental malformations due to premature loss of teeth, fused deciduous teeth, certain dystrophies, submersion of deciduous teeth, dysembryoplasias, and rickets.


[Indication for anesthesia treatment in the oral surgery polyclinic]. On the basis of follow-up examinations in 126 patients, the indications for treatment under general anaesthesia at a maxillofacial clinic are verified, and relevant guidelines are indicated. Oral surgical treatment under general anaesthesia is mainly indicated in mentally defective patients and, if necessary, in over-anxious infants and infants with behaviour disorders.


[Cryotherapy of benign neoplasms of the soft tissues in the maxillofacial area]. In the light of the relevant literature and of personal studies, the author deals with the possibilities of treating benign neoformations in the maxillofacial region by cryotherapy. The results are discussed from the aspects of cryophysical and cryobiological prerequisites. The extension in depth of the changes is the decisive criterion for cold application.


[Otorhinologic and endoscopic findings in cleft palate patients with special reference to velopharyngoplasty]. Our findings (190 patients of whom 45 underwent follow-up examination) permit the conclusion that the velopharyngoplasty according to the principle of Sanvenero-Roselli, as modified by Heiner, does not lead to an increase in tympanic findings (preoperatively, 73%; postoperatively, 71%). The evaluation of the results from tube function testing by means of the pressure difference method reveals an improvement in tuber aeration (preoperatively, 76% of pathological results of the hearing test; postoperatively, 55.6%). This is attributed to a functional improvement of the velopharyngeal mechanisms after velopharyngoplasty. The use of postrhinoscopic nasal endoscopy in the cleft patient and the results obtained are dealth with. The procedure permits to investigate the functional processes in the velopharyngeal region under the conditions of phonation.


[Experience with the advanced training of the dental specialist for general stomatology]. In the urban area of Dresden, the need for continuing education and the programming of the advanced training for the diploma of dentist specialized in general stomatology were analyzed. It was stated that the introduction of the program of advanced training for the diploma of dentist specialized in general stomatology has led to the conceptual and methodological improvement of the educational and training processes. Nevertheless, it became evident that the responsible stomatologists have still to improve the management of the continuing education process.


Engraftment with chronic granulocytic leukemia cells in acute myeloid leukemia. A deliberate engraftment with nonirradiated chronic granulocytic leukemia (CGL) cells was performed in a patient with acute myeloid leukemia (AML) at a time when he was resistant to cytotoxic drug chemotherapy, pancytopenic and developed an infection. The CGL engraftment was confirmed by the presence of a Ph1-positive donor clone in the recipient's bone marrow and by the pattern of colony growth of the recipient's bone-marrow cells cultured in vitro. Bone marrow engraftment in the host helped in the resolution of infection and permitted the administration of further cytotoxic drugs, as a result of which a remission of AML occurred.


Primary renal osteosarcoma.

Primary renal osteosarcoma. A recent case of primary osteosarcoma of the kidney prompted a review of the literature for this rare malignancy. Six cases have been recorded, and this case represents the seventh. The clinical and pathologic features of this lethal tumor are discussed. The presentation of flank pain, gross hematuria, a palpable mass, and a localized parenchymal calcification on x-ray film should alert the clinician to the possibility of renal osteosarcoma.


[Quantitative and electron microscope study of the nerve in seven cases of sporadic idiopathic sensory neuropathy (author's transl)]. An electron microscope study and quantitation of myelinated and unmyelinated fibers of seven nerve biopsies performed in sporadic cases of idiopathic sensory neuropathy is reported. The number of myelinated fibers is markedly decreased or absent in every case. On the contrary, the unmyelinated fiber numbers are normal or increased. In most cases, the small diameter myelinated and unmyelinated fibers proportions are higher than those of control biopsies. The electron microscope study discloses evidence of degeneration of Wallerian tye and regeneration is also indicated by quantitative studies. Regenerative phenomena seen more obvious in sporadic cases than in previously reported studies of familial cases.


Continuous mini-infusion of bupivacaine into the epidural space during labor. Part I: Radiographic visualization of the epidural catheters. Two groups, each consisting of 20 parturients, were given a continuous infusion of 0.25 per cent bupivacaine into the epidural space for pain relief, after test and loading doses. The analgesic effect was registered during labor. After delivery the course of the catheters in the epidural space was investigated radiologically. In the first group long catheters (mean 16.5 cm from the skin surface) and in the second group short catheters (mean 10.5 cm) were used. The medial approach in the epidural space was varied between the first and second lumbar interspaces. The relationship between the position of the catheter tip and the degree of analgesia was also studied. In two mothers with fetus mortuus ante partum, the spread of a radiopaque dye solution was followed during labor. Through the force of gravity and the mothers position, the spread of solution covered three segments within 30 minutes. It was concluded, that slow infusion of local anesthetic solution gives satisfactory pain relief if the catheter is inserted into the first lumbar interspace to a depth of 10--12 cm from the skin surface ending close to the Th12 level. Gradual raising of the mother from the supine to the semirecumbent position and regular change of sides during mini-infusion is important for an even spread of the analgesic solution and analgasia.


Continuous mini-infusion of bupivacaine into the epidural space during labor. Part II: Blood concentration of bupivacaine. Mini-infusion of 0.25 per cent bupivacaine plain into the epidural space was used in 16 healthy women for pain relief during labor. After a loading dose, continuous administration was ensured by using an automatic pump at an infusion rate of 5 or 8 ml per hour. Maternal venous blood concentrations of bupivacaine were determined 20, 40, and 60 minutes after the loading dose. Further samples were taken hourly until delivery. Fetal blood concentrations were determined from fetal scalp blood during mini-infusion, and from umbilical venous blood at delivery. Maternal blood levels averaged 206 ng/ml. The maximum concentration (604 ng/ml) did not exceed 30 per cent of the toxic level. The blood concentration of bupivacaine did not tend to increase during the course of continuous infusion. Tachyphylaxis could not be detected statistically. The fetal blood concentration during blockade averaged 58 ng/ml, giving a feto-maternal ratio of 0.28. The neonatal-maternal ratio, calculated from the umbilical venous blood, was 0.32. With a shift of the acid-base status towards acidosis in the neonate, the ration of bupivacaine slightly increased. Absence of maternal hypotension as well as absence of fetal heart irregularities during infusion may be attributed to the low and stable blood concentration of bupivacaine. The mean Apgar score was 9.5 at one minute and 9.9 at five minutes. The maternal arterial acid-base balance at delivery was normal. The fetal acid-base status at delivery showed wide variation, without any relation to the Apgar score or the total dose of bupivacaine. It is concluded that continuous mini-infusion of bupivacaine by means of an automatic pump has advantages over intermittent administration.


Continuous mini-infusion of bupivacaine into the epidural space during labor. Part III: A clinical study of 225 patients. Obstetric analgesia was accomplished by segmental continuous blockade in 225 women. The technique involved automatic pump infusion of 0.25 per cent bupivacaine solution into the epidural space at a rate of 5 ml per hour after initial doses of 2 and 5 ml bupivacaine. If the analgesia was insufficient one or two single injections of 5 ml of bupivacaine were added. Statistical evaluation of the results could be carried out for 218 women, 158 of whom were nulliparae and 60 multiparae. Fully satisfactory analgesia was achieved in 96 per cent of the nulliparae and 88 per cent of the multiparae in the first stage of labor. In the second stage of labor 46 per cent of the mothers were given pudendal blockade to maintain statisfactory analgesia. The positioning of the patient in the first stage of labor from supine to semirecumbent was of importance to spread the analgesic agent caudally, to the sacral nerve roots, and to control the pain due to stretching of the vagina and perineum. In the total material 17 per cent of the neonates were delivered by vacuum extraction. When the infusion into the epidural space was started in early labor, the incidence of vacuum extraction was 9 per cent, as compared with 38 per cent when it was started at 6 cm cervical dilatation or later (p less than 0.01). 9.8 per cent of the neonates were delivered by cesarean section. Fetal head malposition occurred in 8.7 per cent. A drop in blood pressure was noted in 7 per cent of the women. The condition of the newborn was unaffected by the analgesia. The mini-infusion system minimized the risk for infection. The danger in case of accidental intravascular injection was reduced, due to slowly administered bupivacaine. At the maternity department this technique has created a positive attitude towards epidural blockade, as midwives and doctors have found it safe and easy.


Ripening of the cervix and induction of labor in patients at term by single intracervical application of prostaglandin E2 in viscous gel. Prostaglandin E2 (PGE2) suspended in a viscous gel was deposited intracervically to 115 patients at term with an unripe cervix. 45 of these patients were given a single dose of 1.0 mg PGE2, whereas the remaining 70 received only half that dose, i.e. 0.5 mg. Irrespective of dose greater than 60 per cent of the patients were induced into labor and delivered without further stimulation within less than 24 hours. The mean induction delivery time was 10 hours. In the remaining patients a considerable ripening of the cervix was registered. Hypercontractility did occur in one patient given 1.0 mg PGE2, otherwise no maternal side effects were observed. There were no adverse effects on the fetuses. The number of instrumental deliveries was 19 per cent including 9 per cent cesarean sections. It is concluded that locally applied PGE2-gel can be used to produce ripening of the cervix and/or induce labor in patients at term with an unripe cervix. Since there was practically no difference in efficiency between the two doses but one case of hyperstimulation in patients given 1.0 mg PGE2, the smaller dose, i.e. 0.5 mg is recommended.


Aspects on ripening of the cervix and induction of labor by intracervical application of PGE2 in viscous gel. In this survey the effect of intracervically applied prostaglandin E2 (PGE2) on the pregnant uterus is outlined. It is concluded that PGE2 decreases cervical resistance and stimulates the myometrium to contract. In small doses (0.25 mg and 0.5 mg) PGE2 produces a rapid and prominent ripening of the cervix without systemic side effects and indepedent of prominent uterine contractions. Prostaglandin E2 applied intracervically in viscous gel seems to be superior to previously used methods for treatment of patients with an unfavorable cervical state.


Noise-induced hearing loss in Danish brewery workers. An original computer-fitted diagnosis system for registration of noise-induced hearing loss (NIHL) is presented and compared with the tested persons' subjective hardness of hearing. Among 6 904 brewery workers it was found that people below 40 years only contract little NIHL during the first 5 years of noise exposure. The NIHL after more than 5 years of noise exposure is of the same magnitude as the hearing loss resulting from 10--20 years of age. The NIHL in older age groups was of a minor degree. Permanent use of hearing protectors was associated with good hearing, but the data suggest that good hearing might be the cause and not the effect of the use of hearing protection.


Comparison between stationary and personal noise dose measuring systems. In most of the investigations which have been made to survey occupational noise, a stationary sound level meter has been used either alone, or in combination with a stationary dosimeter. In this investigation, which was performed in a ship-building yard, an ear-borne noise dosimeter has been used. The Leq values which were obtained have been compared with those of the pocket-borne dosimeters. In many processes, considerable differences exist between pocket-borne and ear-borne dosimeters.


Speech intelligibility in and speech interference levels of traffic noise in hearing-impaired and normal listeners. Speech intelligibility (phonetically balanced monosyllables = PB words) in traffic-like noise (random noise) as well as the speech interference levels (75% intelligibility) of this noise were investigated in three common listening situations, two indoors and one outdoors. A total of 279 persons were examined; 56 had normal hearing, 90 a presbycusic-type hearing loss, 95 noise-induced hearing loss, and 38 conductive hearing loss. In the indoor situation the hearing-impaired groups mainly retained their intelligibility in 40 dBA. Lowering of the noise level to less than 40 dBA resulted in only a minor, mostly insignificant improvement in speech intelligibility. Normal-hearing listeners maintained good speech intelligibility in the outdoor listening situation with noise levels up to 60 dBA, without lip-reading. For hearing-impaired groups, representing more than 1/2 million (8% of the population) people in Sweden, with hearing loss due to age and/or noise, the noise level outdoors must be lowered to 45 dBA in order to achieve good speech intelligibility at 1 m, without lip-reading.


Binaural hearing spectacles with "no mould" by acoustic trauma. Patients with selective hearing loss in the trebles, as commonly seen in acoustic trauma, often have a considerable hearing handicap. They seldom benefit from the usual hearing aids with closed ear moulds. When Ewertsen (1970) and Cortois (1972) reported their experiences with binaural fitting of hearing spectacles, using a free plastic tube in the ear canal ("no mould"), we at the Hearing Centre in Namsos began to fit such hearing aids to 33 patients with acoustic traumas. All of them were followed-up in February--March 1978. The results have been evaluated on the basis of answers from the patients entered on questionnaires, and speech audiometry in open field, monosyllable, with 60 dB wide-band background noise. 27 patients (80%) stated that they were greatly satisfied, and 7 (20%) well satisfied. The average daily use was 4 1/2 hrs. All of the patients emphasized the great benefit obtained with the hearing aid at conferences, meetings, social occasions, etc. The average hearing gain was 13 dB. Among those who had discrimination loss there was an average gain of 30% .


The noise protection effect of the stapedius reflex. The purpose was to estimate the attenuation and protection effect of the acoustic stapedius reflex on the inner ear. The acoustic stapedius reflex was recorded by means of extratympanic manometry (ETM). This method was used, as it is able to differ between the stapedius and the tensor reflex. Patients with unilateral Bell's palsy were investigated during palsy time and after recovery. The amplitude of the contralateral acoustic stapedius reflex was used as a relative measure of the excitation of the cochlea on the affected side. The attenuation effect, 20 dB above the reflex threshold, was approximately 15 dB for all tested patients. After the ETM investigation a temporary threshold shift was shown on the affected side (average 34 dB). The hearing threshold on the healthy side was unchanged. Thus the attenuation effect of the acoustic stapedius reflex protects the inner ear from an acoustic damage. A protection effect of the tensor tympani was not demonstrated.


The effect of the stapedius reflex on attenuation and poststimulatory auditory fatigue at different frequencies. The effect of the stapedius reflex on attenuation was measured in patients with unilateral facial palsy (Bell's palsy) and stapedius muscle paralysis. Poststimulatory auditory fatigue was determined in the same patients and in a group of normal test subjects with a normal bilateral stapedius muscle function. The attenuation for a 0.5 kHz tone was found to begin at the reflex threshold, to increase about 7 dB per a 10 dB increase in the stimulus tone and to reach a maximum in the vicinity of 20 dB. When a 2.0 kHz tone was used, no attenuation of note appeared until the sound had risen to about 10 dB above reflex threshold. The attenuation reached a maximum at a little less than 10 dB. A properly functioning stapedius muscle significantly reduced poststimulatory auditory fatigue at 0.75 kHz while at 3.0 kHz, it could do so only if it was stimulated into action by low-frequency noise.


Nasocytologic examination of wood industry workers. To establish a methodology for field examination for early diagnosis of ethmoidal cancer in workers in the furniture industry, two methods were compared, namely nasal rinsing and direct sample-taking from the middle meatus. The latter method proved superior, resulting in a high number of cylinder cells. This method was tried out in a field survey of 715 furniture workers. No tumours of precancerous changes were revealed. The investigation continues, by exposing the Syrian golden hamster to wood-dust and using, amongst other methods, quantitative nuclear morphometry for evaluation.


Occupational etiology and nasal cancer. An internordic project. Reports from Britain and other countries have shown an increased frequency of adenocarcinoma of the nose and paranasal sinuses among workers exposed to fine wood dust. With support from the Nordic Council a joint project has been launched in Denmark, Finland, Norway and Sweden to single out occupations and exposures possibly associated with nasal cancer. The project is a prospective case-referent study with matched control cases of colon cnacer. Each new case of malignant eipthelial tumour of the nose and sinuses reported to the individual national cancer registries is interviewed concerning previous occupational exposures according to the program. The estimated duration of data collection is two years. This type of joint study is new to the Nordic countries and we hope to gain valuable experience in order to improve cooperation within this type of medical research. A retrospective study in Sweden has shown that 50% of all male cases of nasal adenocarcinoma from 1960 to 1972 were joiners, most of them cabinet makers. The duration of exposure to fine wood dust was known in 21 of 22 cases. The latency period varied from 22 to 70 years, with a mean value of 44.7 years. The exposure period varied from 9 to more than 30 years. The results correspond to those reported from Britain and Denmark.


Diagnostic procedures in chronic rhinitis. Various methods of diagnosing allergic factors in chronic rhinitis are discussed. Among the procedures which aim at detecting specific allergens, i.e. skin testing, RAST, and nasal provocation tests, the last mentioned, as they are performed directly on the shock organ, have so far been found to give the most accurate picture of clinically dominant allergens and of the intensity and character of the rhinitis. However, information obtained by analysing the correlations between different procedures is not unanimous. As long as test techniques and allergen extracts have not been standardized, one particular test cannot be recommended as the method of choice.


Mucus production in the nasal sinuses.

Mucus production in the nasal sinuses. The density of goblet cells and glands was determined in the maxillary, ethmoidal, frontal, and sphenoidal sinuses. Goblet cells showed the highest density in the maxillary sinuses. 9700 per mm2, while in the other sinuses their density was approximately the same, from 5 900 to 6 500 cell per mm2. The density of glands was highest in the ethmoids, 0.5 gland/mm2, then in the maxillary sinuses, viz. 0.2 gland/mm2, while in the frontal and sphenoidal sinuses it was very low, 0.08 and 0.06 gland/mm2 respectively.


A population study of otitis media in childhood. A total of 494 children with an average age of 3 1/2 years were studied with a view to elucidating the frequency of both acute and secretory otitis media and their relationship, if any, to a number of social factors. Of the children, 41% had had at least one attack of otitis media, in 49% of these associated with aural discharge. 70% of the children were attending day-care centres, and the incidence of otitis media was highest in this group. According to Jerger's classification of the tympanometric curves, 51% of these were of type A, 17% of type B, and 32% of type C. Significantly higher frequencies of types B and C curves were found among children attending day-care centres.


Inflammatory middle ear diseases in patients with cystic fibrosis. 111 patients with cystic fibrosis (CF), aged 7 months to 29 years, were examined to detect inflammatory middle ear diseases. 35% had a history of one or more attacks of acute otitis media, an incidence at the same level as in non-CF patients. None had chronic suppurative otitis. Only 4 out of the 88 patients able to cooperate in audiometry had a hearing impairment. In 2, the hearing loss was perceptive, in 3 very mild, and in the fourth case, moderately severe. The middle ear pressure was measured in 108 patients, in 86 of whom it proved normal. In 15 patients the middle ear pressure was lower than--100 mmH2O, indicating tubal occlusion, and in 7 patients the impedance minimum was lacking, indicating an accumulation of mucus. These 7 patients (6%) had secretory otitis media. Nasal polyps were or had been present in 32%. There was no correlation between nasal polyps and tubal occlusion or secretory otitis. The incidence of inflammatory middle ear diseases in CF patients was in the same range as in non-CF patients, a finding at variance with some previous investigations showing middle ear pathology in 25--48%.


Middle ear mechanics and Eustachian tube function in tympanoplasty. In 100 patients with chronic otitis media, tubal function and volume was tested before operation. In a postoperative study on hearing and healing there was no positive correlation between these findings and the preoperative results. In 50 patients with healed ear drums, tubal function was retested in a pressure chamber with a flow volume technique. An improvement in Eustachian tube function could be demonstrated. These findings indicate that poor tubal function in chronic otitis media might be secondary to other factors responsible for the disease.