Production and characterization of a monoclonal antibody to chicken type I collagen. We have shown that lymphocyte-myeloma cell hybridization can be used to produce large amounts of extremely high-titer specific antibodies against type I collagen, a macromolecule normally of low immunogenicity. In a passive hemagglutination assay the antibody had a high titer against chicken type I collagen but showed no activity against chicken type II or rat type I collagen. By using a two-step fluorescence histochemical procedure on sections of embryonic chicken tibia, strong fluorescence was observed in the perichondrium and surrounding connective tissue (known to contain type I collagen) but not over the cartilage (characterized by type II collagen). When used in conjunction with Staphylococcus aureus as a solid phase immunoadsorbant, the antibody was shown to bind to labeled collagen synthesized in vitro by embryonic chicken calvaria.


Human leukocyte interferon: relationship between molecular structure and species specificity. Human leukocyte interferon can be separated into two classes of subspecies by polynucleotide-agarose affinity chromatography; 30-40% of the molecular species have the polynucleotide-binding property and 60-70% lack affinity for the polynucleotide ligand. When analyzed on sodium dodecyl sulfate/polyacrylamide gel electrophoresis, the former class of interferon has a slower mobility corresponding to the migration of a polypeptide of 21,000 daltons, while the latter class has a faster mobility corresponding to a polypeptide of 13,500-15,000 daltons. By analogy to the behavior of other interferons and a class of nucleotidyl transferases on the polynucleotide-agarose chromatography, we suggest that the human leukocyte interferon having the polynucleotide-binding site is in a possibly "native" conformation and the loss of affinity for polynucleotide results from a degradative alteration of the native molecules. Moreover, the alteration of interferon is accompanied by an increase in heterospecific activity on bovine cells. It is suggested that the polypeptide domain responsible for species specificity may be closely related to the polynucleotide binding area. The modified interferon molecule, however, still conserves its antiviral activity. The simplicity and the high capacity of polynucleotide-agarose chromatography make this a powerful technique for the purification of interferon. The easy separation of these two classes of human leukocyte interferon makes the purification procedures more rational and will facilitate the preparation of both subspecies to a high degree of molecular homogeneity.


Platelet-derived growth factor: purification and partial characterization. A cationic protein that stimulates DNA synthesis in human cultured cells was isolated from human platelets by ion exchange chromatography, hydrophobic chromatography, gel chromatography, and gel electrophoresis in sodium dodecyl sulfate. The electrophoretic behavior of biologically active or radioiodinated and reduced growth factor indicated that the native protein (approximately 30,000 daltons) was composed of two different polypeptides (approximately 13,000-14,000 and 16,000-17,000 daltons, respectively) linked via reduction-susceptible bonds. The stimulatory activity on human glial cells of the purified product at a concentration of approximately 4 ng/ml (0.13 nM) was equal to that of 1% human serum.


Covalently bound non-coenzyme phosphorus residues in flavoproteins: 31P nuclear magnetic resonance studies of Azotobacter flavodoxin. In addition to the 5'-phosphate ester on its flavin mononucleotide (FMN) moiety, flavodoxin from Azotobacter vinelandii contains 2 moles of tightly bound phosphate. One non-coenzyme phosphate group is covalently bound to the protein, as it remains with the protein on acid precipitation, whereas the other phosphate is released. The invariance of the (31)P nuclear magnetic resonance chemical shift of the covalently bound phosphate (-0.8 ppm relative to 85% phosphoric acid) with pH, even in the presence of protein denaturants, implies it is in a diester linkage to the protein. Because no evidence could be found for the presence of covalently bound sugars, nucleotides, or phospholipids, it is suggested that the phosphate residue forms a diester linkage with two hydroxyl amino acids in the protein. The only other suggestion of a phosphodiester linkage in proteins is from previous studies on pepsin and pepsinogen [Perlmann, G. E. (1955) Adv. Prot. Chem. 10, 1-30]. The observed changes in (31)P chemical shift with pH show that the covalent phosphorus in pepsinogen has ionization properties of a monoester rather than a diester. The (31)P resonance of the FMN phosphate occurs at -5.6 ppm in native Azotobacter flavodoxin. No ionization of the protein-bound FMN phosphate is observed since the chemical shift does not change appreciably in the pH range of 5.5-9.5. The chemical shift data suggest, but do not prove, that the coenzyme phosphate in its protein-bound form is dianionic. Chemical analysis of several other flavoenzymes from a variety of sources shows the presence of covalently bound phosphorus in quantities stoichiometric with the flavin content in most of the enzymes tested. Thus, the presence of covalent phosphorus in flavoenzymes may be a general phenomenon with currently unknown catalytic significance.


The smallest genome RNA segment of influenza virus contains two genes that may overlap. The genome of influenza virus consists of eight segments of single-stranded RNA, each of which encodes a different polypeptide. In addition to the eight recognized gene products, the virus specifies a distinct smaller nonstructural polypeptide (NS2), which is translated from a separate species of virus-specific mRNA. The location on the virus genome of the gene encoding this polypeptide was investigated by hybridization of the NS2 mRNA with isolated subgenomic RNA species, and by correlation of the inheritance of a strain-specific NS2 with inheritance of particular genome RNA segments during recombination between two different virus strains. The genetic information for NS2 was found to reside in the smallest genome RNA segment of the virion, which also encodes the NS1 polypeptide. Considering the sizes of the molecules involved, it is likely that the coding sequences for the two polypeptides overlap.


Immunological and chemical identification of a neurophysin-containing protein coded by messenger RNA from bovine hypothalamus. The biosynthetic origin of the 10,000 molecular weight neurophysins, carriers of the peptide hormones oxytocin and vasopressin, has been studied by cell-free synthesis, Poly(A)-RNA was isolated from bovine hypothalamus and translated in a wheat germ system containing (35)S- or (3)H-labeled amino acids. A number of unique [(35)S]cysteine- but few [(35)S]-methionine-labeled proteins were coded by hypothalamic mRNA. A single, major, isotopically labeled protein (molecular weight 23,000-25,000) was immunoprecipitated from these translation mixtures by addition of purified antibodies against bovine neurophysin II and subsequent addition of Cowan I strain of Staphylococcus aureus. Specificity of the immunoprecipitation was demonstrated by competition with unlabeled authentic neurophysins and the absence of competition with structurally unrelated ovalbumin. Furthermore, neither nonimmune serum nor purified antibodies against ribonuclease immunoprecipitated the protein. The [(35)S]cysteine-labeled protein that was specifically immunoprecipitated was oxidized with performic acid and digested with trypsin in the presence of unlabeled, authentic bovine neurophysin II. Peptide mapping revealed that most of the major [(35)S]cysteine-labeled peptides (of the translation product) were identical to major cysteine-containing peptides of authentic neurophysin. The data show that hypothalamic mRNA directs the translation of several unique cysteine-rich proteins in an in vitro cell-free system. Furthermore, one of these proteins, which has a higher molecular weight than authentic neurophysin, is recognized by purified antibodies to bovine neurophysin II and has cysteine-containing tryptic peptides in common with those of authentic neurophysin. The data suggest that this protein is the primary translation product, pre-pro-neurophysin.


Identification of separate mRNAs coding for the alpha and beta subunits of thyrotropin. A 9S mRNA, purified from mouse thyrotropic pituitary tumors by sucrose density gradient centrifugation of poly(A)-enriched mRNA, directed the synthesis of only alpha and beta subunits of thyrotropin in the reticulocyte lysate translation system. Analysis of radioiodinated 9S mRNA, repurified by oligo(dT)-cellulose chromatography and sucrose gradient centrifugation, yielded two species of RNA on urea/polyacrylamide gel electrophoresis. The major RNA species contained 620 nucleotides, and the minor RNA species contained 560 nucleotides. Unlabeled 9S mRNA was further purified by urea/polyacrylamide gel electrophoresis; the mRNAs were separately eluted from slices of the gel containing material migrating with an apparent length of 620 and 560 nucleotides. Translation of these mRNAs in the reticulocyte lysate showed that the longer mRNA coded for the alpha subunit and the shorter mRNA coded for the beta subunit of mouse thyrotropin. Because more alpha than beta subunit of thyrotropin was consistently synthesized, unbalanced amounts of thyrotropin subunits appear to be synthesized by translation of unbalanced amounts of individual mRNAs. We have demonstrated that the synthesis of thyrotropin is directed by two separate mRNA molecules, each coding for a different subunit of the hormone.


In vivo distribution and turnover of fluorescently labeled actin microinjected into human fibroblasts. Graessmann's microinjection technique was chosen to introduce fluorescently labeled muscle actin and other proteins into WI-38 human fibroblasts. The injected cells were examined during culture by fluorescence and reflection contrast microscopy. Within 30 min after injection, rhodamine-labeled actin was incorporated into a distinct network of fluorescent filaments, resembling the stress fibers observed by classical immunofluorescence microscopy. Cytochalasin b prevented the formation of these fibers, but colchicine did not. Neighboring fibers often converged into distinct focal points that appeared to be concentrated near the base of the cell. Examination of these fluorescent fibers and focal points by reflection contrast microscopy confirmed their close location to the substratum. After 6 hr of culture, fluorescent actin and the control proteins were segregated into granules located mainly near the cell nucleus. Thus, the injected actin both enters the intrinsic actin pool and participates in an assembly and disassembly of filamentous structures. Segregation into granules traces the natural turnover of this protein within the cell.


Primary structure of the NH2-terminal extra piece of the precursor to human placental lactogen. The cell-free translation product of human placental lactogen mRNA is a precursor molecule larger than the mature hormone that circulates in plasma. To determine the structure of pre-placental lactogen, the poly(A)-rich RNA fraction of term placenta was isolated and translated in a wheat germ cell-free system. The mRNA programmed the synthesis of a major protein, 3000 daltons larger than placental lactogen, that was specifically precipitated by hormone antibodies. The immunoprecipitated protein was labeled separately with 20 radioactive amino acids and subjected to sequence analysis. The results showed the synthesis of pre-placental lactogen in which an extra piece 25 residues long preceded the NH2 terminus of the mature protein. The structure of the extra piece is as follows: Met-Pro-Gly-Ser-Arg-Thr-Ser-Leu-Leu-Ala-Phe-Ala-Leu-Leu-Cys-Leu-Pro-Trp-Leu-Gln-Glu-Ala-Gly-Ala-. Met1 is the initiator residue because only initiator [35S]Met-tRNAMet1, but not internal [35S]Met-tRNA2Met, donated NH2-terminal methionine. The structure of the extra piece showed little homology with that of unrelated hormones but striking homology (64%) with the extra piece of rat pre-growth hormone. Most amino acid substitutions involved a single base change in the codon. Mature human placental lactogen and rat growth hormone have 59% homology in sequence. Thus, our findings provide additional evidence to support the common evolutionary origin of these hormones, not only of the mature proteins but also of the extra piece segments.


Quantum chemical calculations of model systems for ascorbic acid adducts with Schiff bases of lysine side chains: possibility of internal charge transfer in proteins. Ab initio self-consistent field calculations for neutral and cationic ascorbic acid and model compounds have been performed. Furthermore, the bicyclic addition products of alpha-hydroxytetronic acid with methylglyoxal and with the Schiff base formed between methylglyoxal and methylamine have been calculated, again in their neutral and cationic forms, respectively. The results indicate that the investigated cations can act as strong electron acceptors. With the help of space-filling molecular models it has been demonstrated that such conformations of the Schiff base formed between the primary amino group of lysine side chains in proteins and the ascorbic acid methylglyoxal acetal are possible in which the lactone carbonyl group comes near to the N atoms of the peptide groups, so that charge can be transferred between these subunits.


Pore formation in lipid membranes by alamethicin. The conformation of the linear peptide antibiotic alamethicin in dipalmitoyl phosphatidylcholine multilayers was investigated in the absence of an electric field by means of infrared attenuated total reflection spectroscopy. Alamethicin was found to be incorporated into the lipid membrane not only in the dry state but also in an aqueous environment. Its molecular conformation, however, changed from a helix when dry to an extended chain when aqueous. The extended chain aggregated to di- and multimers spanning the lipid bilayer. The equilibrium concentration of alamethicin in the surrounding water was 90 nM, which is in the range of concentrations used in black film experiments. The corresponding molar ratio of lipid to peptide was 80:1. Concerning the molecular mechanism of electric field-induced pore formation, one has to conclude that the dipole model proposed by several authors is very unlikely because it is based on the assumption that the major part of alamethicin is adsorbed on the membrane surface, from which small amounts flip into the membrane under the influence of an electric field. An alternative mechanism is proposed, based on a field-induced conformational change of the peptide from the extended state to a helix. This transition is favored by the resulting dipole moment of the alamethicin helix.


On the molecular basis for chemomechanical energy transduction in muscle. Herein it is developed that energy transduction in muscle is an activity of myosin S-1 and its ligands, actin (A) and nucleotide (N). S-1 shares with other molecular particles (e.g., hemoglobin) the property that binding events at one of its sites, the N-site, influences binding events at a remote site, the A site (specifically, influences both the actin affinity and actin attachment angle at the A site). However, there is a crucial difference between S-1 and the better-known systems. Because the N site is enzymatic, it has a temporal sequence of occupants; this imposes a temporal sequence of actin attitudes--i.e., a sequence of mechanical events.


Coexistence of desmin and the fibroblastic intermediate filament subunit in muscle and nonmuscle cells: identification and comparative peptide analysis. Extraction of chicken embryo fibroblasts (CEF) or baby hamster kidney (BHK) cells with 1% Triton X-100 and 0.6 M KCl leaves an insoluble cytoskeletal residue composed primarily of the 52,000 Mr subunit of intermediate filaments (F-IFP). In addition, CEF cytoskeletons exhibit a minor component with Mr of 50,000, identified as alpha-desmin, one of the two major isoelectric variants of the intermediate filament subunit from smooth muscle. BHK cytoskeletons contain the 50,000 Mr mammalian desmin variant. Cytoskeletons prepared from chicken embryonic myotubes contain F-IFP and both alpha- and beta-desmin. These data suggest that two distinct 10-nm filament subunits coexist in a single cell. One-dimensional peptide analysis of F-IFP and desmin from avian and mammalian cells reveals significant interspecies homology, as well as homology between F-IFP and desmin from the same species. Peptide analyses of 32P-labeled intermediate filament subunits suggest that there is considerable similarity in the phosphorylation sites of these proteins. These results indicate that F-IFP and desmin might be evolutionally related.


Modulation of plasminogen activator secretion by activated macrophages: influence of serum factors and correlation with tumoricidal potential. A high molecular weight fraction of human serum (Fr-1) was found to both inhibit macrophage tumoricidal activity and enhance plasminogen activator activity in supernates over activated macrophages in vitro. Conversely, a 40- to 90-kilodalton serine esterase (Fr-3) also found in normal human serum and endotoxin enhanced tumoricidal potential and suppressed the supernatant plasminogen activator activity. Inactivation of either Fr-1 or Fr-3 by 2-mercaptoethanol or diisopropyl fluorophosphate, respectively, abolished both biologic effects. Examination of cell-associated and culture medium plasminogen activator activity before and after acidification to inactivate proteinase inhibitors indicated that suppression of plasminogen activator activity by Fr-3 or endotoxin most likely represents modulation of macrophage plasminogen activator secretion. The findings demonstrate that activated macrophages are capable of highly coordinated biologic responses to alterations in their microenvironment and suggest that it is in fact the high potential for such responsiveness that reliably characterizes the activated macrophage. The results also suggest that an endogenous regulatory system dependent on the interaction of serine esterases may operate to regulate the functional capabilities of activated macrophages.


Major intracellular cations and growth control: correspondence among magnesium content, protein synthesis, and the onset of DNA synthesis in BALB/c3T3 cells. Omission of Ca(2+) from the medium of confluent BALB/c3T3 cells for a period of 17 hr causes a large decrease in the number of cells synthesizing DNA. This effect is reversed by raising the Mg(2+) concentration of the medium to 20 mM. However, if the [Mg(2+)] is greater than 20 mM ("ultra-high" Mg(2+)), there is again a decrease in the number of cells synthesizing DNA. The synthesis of protein has a similar dependence on Mg(2+) concentration in Ca(2+)-deficient medium, but it responds within 45 min of the shift in cation concentrations rather than the 10 hr that is required for the change in DNA synthesis to become apparent. Cells in the ultrahigh Mg(2+) concentrations that are at first inhibitory to protein synthesis later return to maximal protein synthesis. This delayed increase in protein synthesis is reflected in a delayed increase in DNA synthesis. Intracellular concentrations of Mg(2+) in Ca(2+)-deficient media increase in proportion to extracellular Mg(2+) concentrations. Cells in medium with 30 mM Mg(2+) have a high intracellular content of Mg(2+) at 3 hr but have decreased their intracellular content by 17 hr, a time at which protein synthesis has been restored to normal. Intracellular Na(+) and K(+) concentrations also change in Ca(2+)-deficient medium, but independent variation of these ions shows that protein synthesis is relatively insensitive to their concentration. Intracellular Ca(2+) remains fairly constant under all these conditions. The rate of protein synthesis of intact cells changes as a function of intracellular Mg(2+) content in a manner very similar to that which has been reported for cell-free systems. We conclude that protein synthesis is very sensitive to small changes in intracellular [Mg(2+)] within physiological range and that the onset of DNA synthesis is dependent on the rate of protein synthesis. Regulation of the availability of Mg(2+) within the cell therefore presents a plausible mechanism for growth control.


Bioenergetic pattern of turtle brain and resistance to profound loss of mitochondrial ATP generation. The adaptations in the freshwater turtle that permit survival despite prolonged loss of mitochondrial ATP generation were investigated by comparing the bioenergetics of turtle brain slices with rat brain slices. Aerobic turtle brain shows no significant difference in basal levels of total ATP generation compared to rat brain; levels in turtle brain and rat brain were 18.4 +/- 2.8 (SD) and 19.4 +/- 2.2 mumol (100 mg of tissue)-1 hr-1, respectively. However, in turtle brain, a significantly greater fraction of ATP is derived from glycolysis both under aerobic and anaerobic conditions [aerobic turtle (24%) and rat (13%), P less than 0.02; anaerobic, turtle (28%) and rat (18%), P less than 0.05]. The increased glycolytic capacity is related to high levels of rate-limiting glycolytic enzymes, such as pyruvate kinase (EC 2.7.1.40). Turtle brain operates close to glycolytic capacity even under aerobic conditions, and no Pasteur effect can be demonstrated. Quantitatively, anaerobic glycolysis accounts for a maximum of 28% of basal aerobic ATP generation, suggesting that prolonged diving is also accompanied by a reduction in brain energy requirements. The adaptation subserving short-term (natural) diving is an increase in brain glycolytic capacity. The adaptation subserving prolonged diving (days to weeks) may be a reduction in the energy requirements of brain (and other cells).


Postreplication repair: questions of its definition and possible alteration in xeroderma pigmentosum cell strains. DNA synthesis in normal cells and in excision-defective and variant xeroderma pigmentosum cells was investigated after irradiation with ultraviolet light. The sizes of DNA synthesized during brief pulses of [3H]thymidine 1-2 hr after irradiation were decreased, the xeroderma pigmentosum variant showing the smallest molecular weight. Once synthesized, however, labeled DNA increased in size at the same rat as control in all cell strains, and the rate was relatively insensitive to caffeine. After 2-3 hr, labeled DNA in each cell type reached a maximum size that was less than that in control cells, indicating the presence of long-lived blocks to DNA chain growth. This kind of experiment (pulse-chase) has in the past been used to investigate a repair process believed to be associated with the bypass of damaged sites in parental DNA: postreplication repair. We present an alternative model that does not involve a specific postreplication repair mechanism, but involve a specific postreplication repair mechanism, but involves normal chain elongation and termination mechanisms in which we conceive that dimers and other damaged sites act as well-or-nothing blocks to the progress of replication forks. No evidence could be found for any inducible process that enhanced the bypass of damaged sites.


Epigenetic activation of phenylalanine hydroxylase in mouse erythroleukemia cells by the cytoplast of rat hepatoma cells. Friend mouse erythroleukemia cells do not synthesize detectable levels of phenylalanine hydroxylase [phenylalanine 4-monooxygenase; L-phenylalanine, tetrahydropteridine:oxygen oxidoreductase (4-hydroxylating), EC 1.14.16.1] and hence are unable to grow in medium totally lacking tyrosine. These cells were fused with the cytoplasts of rat hepatoma cells that synthesize phenylalanine hydroxylase constitutively. Cytoplasmic hybrids [cybrids, Bunn, C. L., Douglas, C. W. & Eisenstadt, J. M. (1974) Proc. Natl. Acad. Sci. USA 71, 1681--1685] were selecte in medium without tyrosine. Cybrid clones expressed phenylalanine hydroxylase enzyme, which was of mouse type as determined by immunotitration and isoelectric focusing. This phenotype has been mainta ined even in the absence of any selective pressure. In contrast, in whole cell hybrids derived between the same parents, the expression of the phenylalanine hydroxylase gene was totally extinguished. One interpretation of these results is that the cytoplasm of rat hepatoma cells contain a positively acting factor(s) for the phenylalanine hydroxylase gene that brings about the activation of this gene in erythroleukemia cells.


Isolation and characterization of germ line DNA from mouse sperm. Mouse germ line DNA was isolated from sperm by a physicochemical procedure that preferentially destroys contaminating somatic cell DNA. The use of reducing conditions and chelating agents in combination with phenol permitted extraction of molecular weight DNA from mature sperm nuclei with approximately 80% efficiency. Less than 0.1% somatic cell DNA contamination remained in sperm DNA prepared by this method. Germ line DNA was characterized by determination of its ultraviolet absorbance spectrum, buoyant density in cesium chloride, and melting profile on a hydroxyapatite column. Contamination by mitochondrial DNA was assessed by cesium chloride/ethidium bromide gradient centrifugation. The significance of the mouse germ line DNA isolation procedure is discussed with respect to the possible genetic transmission of mammary tumor virus and leukemia virus, the origin of antibody diversity, and the origin of testicular teratomas.


Multiple phosphorylation of ribosomal protein S6 during transition of quiescent 3T3 cells into early G1, and cellular compartmentalization of the phosphate donor. At 5 min after quiescent cells are induced to enter G1 there is a large increase in the amount of 32P incorporated into 40S ribosomal protein S6. Here we show that changes in the specific activities of 32Pi and [gamma-32P]ATP in stimulated as compared to quiescent cultures do not account for this large increase. Instead, we demonstrate by decreased electrophoretic mobility on two-dimensional polyacrylamide gels that this increase is due to a quantitative increase in the total amount of phosphate incorporated into S6. Furthermore, pulse-chase experiments show that the phosphate that is incorporated into S6 is metabolically stable during at least the first 60 min of induction and that the incorporation of 32P into S6 responds immediately to the replacement of 32Pi by Pi in the medium, in contrast to [gamma-32P]ATP which changes very slowly. Thus, the S6 phosphate donor must be a compartment separate from that of the total cellular ATP.


Transcription of spacer sequences in genes coding for ribosomal RNA in Xenopus cells. Untreated Xenopus cells synthesize RNA is increased and some sequences complementary to those of the nontranscribed spacer are found in the heavy shoulder of the 40S precursor rRNA peak. In such events transcription initiation seems to take place within the spacer because its middle and right BamHI endonuclease fragments are preferentially transcribed, whereas only few RNA sequences complementary to the left spacer fragment are found. It is concluded that at least some spacer regions contain promoters for transcription and can be transcribed either into a special class of "spacer transcripts" or into molecules covalently linked to rRNA precursor.


Microinjection of fluorescently labeled alpha-actinin into living fibroblasts. alpha-Actinin from chicken gizzard labeled with tetramethylrhodamine isothiocyanate has been incorporated into living fibroblast cells by microinjection. Fluorescent labeling of alpha-actinin was carried out such that the conjugated protein was functional in vitro as shown by its ability to bind to F-actin. Within 1-2 hr after injection, diffuse fluorescence was observed throughout the cytoplasm and only faint fluorescence was apparently associated with the stress fibers. During the ensuing 2-15 hr, however, most of the fluorescence was seen as periodicities along the stress fibers and as foci of the microfilament polygonal networks. This distribution of alpha-actinin in the living cells was strikingly similar to that found by indirect immunofluorescence localization of endogenous alpha-actinin in fixed samples of the same cell type. Control studies in which heat-treated (100 degrees C, 2 min) fluorescent alpha-actinin or tetramethylrhodamine isothiocyanate alone was injected into the cells indicated that the stress fiber and polygonal network labeling was specific for "native" fluorescently labeled alpha-actinin. These results suggest that the dynamic properties of proteins and structures in cultured mammalian cells can be studied with the use of microinjection and fluorescence microscopic techniques.


Segregation of pre-stalk and pre-spore cells of Dictyostelium discoideum: observations consistent with selective cell cohesion. Aggregating Dictyostelium discoideum cells were dissociated and separated into two populations, pre-stalk and pre-spore cells, by centrifugation in a Percoll gradient. After harvesting, one population of cells was labeled with fluorescein isothiocyanate and the other with tetramethylrhodamine isothiocyanate. Equal numbers of the two cell populations were then mixed and observed while they aggregated and differentiated to form fruiting bodies. Within minutes after mixing, the two populations showed evidence of segregation and tended to form separate streams of cells as they migrated toward aggregation centers. Many separate clusters of tetramethylrhodamine isothiocyanate-labeled or fluorescein isothiocyanate-labeled cells were found within an aggregation center. Thereafter there was further segregation. The denser cells from the gradient tended to assume the position of stalk cells whereas the lighter cells migrated to the position characteristic of spore cells. The patterns of segregation suggest that selective cohesiveness of one or both cell types plays a role in morphogenesis.


Ultraviolet light induction of diphtheria toxin-resistant mutants of normal and xeroderma pigmentosum human fibroblasts. The UV induction of diphtheria toxin-resistant (DTr) mutants in normal and xeroderma pigmentosum human fibroblasts has been quantitatively characterized. A concentration of diphtheria toxin at which DTr cells are cross-resistant to Pseudomonas aeruginosa exotoxin A was determined and used in the selection of resistant mutants. Recovery of mutants was not influenced by the presence of wild-type cell densities of 1-8 x 10(5) per 9-cm plate, indicating no metabolic cooperation exists, in contrast to what is seen in the selection of some other variant phenotypes. Expression periods for UV-induced mutations differed with the severity of mutagen treatment and cell strain used. A relatively long (10-15 days after UV treatment) expression period was required for the maximum recovery of DTr mutants. Maximum recovery was followed by a decrease in mutation frequency on subsequent days evaluated. An apparent linear dose response within the dose range used was observed for UV-induced mutations in both normal and xeroderma pigmentosum fibroblasts. Our results indicate that xeroderma pigmentosum fibroblasts have higher UV-induced mutation frequencies per unit UV dose but similar frequencies per unit survival compared to normal cells within the range of UV doses tested.


Origins of metabolic diversity: evolutionary divergence by sequence repetition. Recurring patterns of primary structure have been observed in enzymes that mediate sequential metabolic reactions in bacteria. The enzymes, muconolactone Delta-isomerase [(+)-4-hydroxy-4-carboxymethylisocrotonolactone Delta(2)-Delta(3)-isomerase, EC 5.3.3.4] and beta-ketoadipate enol-lactone hydrolase [4-carboxymethylbut-3-enolide(1,4)enol-lactone-hydrolase, EC 3.1.1.24], have been coselected in bacterial populations because the isomerase can confer no nutritional advantage in the absence of the hydrolase. Similar amino acid sequences recur within the structure of the isomerase, and the amino-terminal amino acid sequence of the isomerase from Pseudomonas putida appears to be evolutionarily homologous with the corresponding sequence of a beta-ketoadipate enol-lactone hydrolase from Acinetobacter calcoaceticus. One interpretation of the sequence repetitions is that they reflect tandem duplication mutations that took place early in the evolution of the proteins. According to this view, the mutations caused elongation of structural genes and the creation of duplicated genes as the metabolic pathways evolved. A review of the sequence data calls attention to a different hypothesis: repeated amino acid sequences were introduced in the course of the proteins' evolution by substitution of copies of DNA sequences into structural genes. Our observations are interpreted on the basis of a model proposing genetic exchange between misaligned DNA sequences. The model predicts that misalignments in one chromosomal region can influence the nature of mutations in another region. Thus, as often has been observed, the mutability of a base pair will be determined by its location in a DNA sequence. Furthermore, the intrachromosomal recombination of DNA sequences may account for complex genetic modifications that occur as new pathways evolve. The model provides an interpretation of an apparent paradox, the rapid creation of new metabolic traits by bacterial genomes that are remarkably resistant to genetic drift.


Population genetics of multigene family with special reference to decrease of genetic correlation with distance between gene members on a chromosome. A mathematical method is developed which enables us to treat exactly the process of coincidental evolution under mutation, unequal intrachromosomal crossing-over as well as ordinary crossing-over between homologous chromosomes in a finite population of the effective size N. It makes use of finite difference equations involving two quantities denoted by fi and phi i, in which fi is the identity coefficient of two gene members that are i steps apart on the same chromosome and phi i is that of two members i steps apart on two homologous chromosomes. When the number of genes (n) per family is large, the finite difference equations can be approximately by ordinary second-order differential equations which can then be solved analytically. Results obtained by the present method are compared with the corresponding results previously obtained by one of us (T.O.) using conventional diffusion models of gene frequency changes in population genetics. It is shown that the previous results obtained by T.O. regarding second-order statistics are essentially valid, and they give good approximations particularly when N beta is small, where beta is the rate of ordinary interchromosomal crossing-over within the multigene family.


Human cell lines expressing intestinal alkaline phosphatase. At least three loci determine human alkaline phosphatases [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1]: one coding for the placental form of the enzyme, at least one coding for the intestinal forms, and at least one for the liver, bone, and kidney forms. The alkaline phosphatase in cell line D98/AH-2 has been characterized by inhibition, thermostability, and electrophoretic studies. It is intestinal in type and resembles the fetal intestinal form somewhat more closely than the adult intestinal form. Intestinal alkaline phosphatase was found in the related cell lines Detroit 98, D98/S, and D98/AH-R. No placental alkaline phosphatase could be detected in any of these cell lines. This series of cell lines are believed, on the basis of earlier investigations, to be HeLa in origin but other HeLa cell lines show placental alkaline phosphatase. Loss of expression of the placental alkaline phosphatase locus probably occurred prior to the separation of Detroit-98 from the lineage leading to other HeLa cell lines and this has persisted in the Detroit-98 derivatives D98/AH-2, D98/S, and D98/AH-R. Another possibility is that placental alkaline phosphatase expression only appeared in the HeLa lineage subsequent to the separation of Detroit-98.


Crystal structure of galactan-binding mouse immunoglobulin J539 Fab at 4.5-A resolution. An electron-density map of the mouse galactan-binding immunoglobulin J539 (IgA2,kappa) Fab has been calculated to a resolution of 4.5 A by the method of heavy atom isomorphous replacement with four derivatives. The map has been interpreted with the aid of a computer program which systematically searched for the best fit between the electron-density map and the known coordinates of individual immunoglobulin domains. The quaternary structure of J539 Fab at this resolution appears similar to that of another mouse immunoglobulin, IgA2,kappa Fab, McPC603. The model coordinates for J539 Fab should allow us to proceed directly to a high-resolution structure determination without further heavy atom isomorphous replacement.


Evaluation of a common receptor-complement concept for predicting the antileukemic activity of nucleosides. The triangulation arrangement of one nitrogen and two oxygen atoms proposed as a requirement for antileukemic activity involving a "common receptor-complement feature" has been reevaluated specifically as it applies to nucleosides. This re-evaluation has resulted in a rejection of the "receptor-complement feature" as a working model for the a priori design of nucleosides as compounds active against leukemia L-1210.


Induction of indoleamine 2,3-dioxygenase in mouse lung during virus infection. Indoleamine 2,3-dioxygenase [indoleamine: oxygen 2,3-oxidoreductase (decyclizing)] activity in the supernatant fraction (30,000 X g, 30 min) of mouse lung homogenate increased approximately 120-fold after infection with PR8 influenza virus. Both specific and total enzyme activities started to increase linearly from the 5th day after infection, reached the highest level around the 11th day, and then gradually decreased to normal values in about 3 weeks. Other enzymes in the lung, such as certain lysosomal enzymes and monoamine oxidase, did not change significantly throughout the experiments. The time course of the increase in the enzyme activity was quite different from that of virus replication in the lung (a peak by the 3rd day and persistence until the 9th day) or that of serum antibody content (started to rise on the 9th day). Rather, it appeared to be closely related to the infiltrations of mononuclear and lymphocytic cells. When mice were exposed to a higher dose of virus and did not recuperate, the time course of the increase of the enzyme activity was essentially identical to that seen with a low concentration of virus. A maximum stimulation of the enzyme activity in the lung occurred on the 9th day after infection; the increase was approximately 100-fold. However, serum antibody content was slight and virus titer in the lung remained high.


Specific changes in the surface glycoprotein pattern of human promyelocytic leukemic cell line HL-60 during morphologic and functional differentiation. The human promyelocytic leukemia cell line HL-60 can be induced to undergo morphological and functional differentiation in vitro by various low molecular weight compounds. The cellular morphology changes from blastoid appearance to that of granulocytes and the cells acquire the ability to phagocytize. We here report that the surface glycoproteins specifically change during this differentiation, as shown by the neuraminidase/galactose oxidase/NaB3H4 surface-labeling technique followed by polyacrylamide slab gel electrophoresis. The most prominent change is the loss of the major glycoprotein band typical for the blast cells which has an apparent molecular weight of 160,000 and the appearance of a major surface glycoprotein band with an apparent molecular weight of 130,000. Expression of the 130,000 molecular weight band correlates with the appearance of phagocytic and chemotactic activities of the cells. It has the same molecular weight as the major surface glycoprotein of freshly isolated human blood granulocytes.


13-Azaprostanoic acid: a specific antagonist of the human blood platelet thromboxane/endoperoxide receptor. A newly synthesized 13-aza derivative of prostanoic acid (13-APA) specifically inhibited human platelet aggregation induced by arachidonic acid, prostaglandin H2, or the stable endoperoxide analog (15S)-hydroxy-9 alpha,11 alpha-)epoxymethano)-prosta-5Z,13E-dienoic acid. 13-APA also inhibited [14C]serotonin release in response to arachidonic acid, ADP, or thrombin, but did not inhibit primary aggregation induced by ADP or thrombin. 13-APA completely blocked prostaglandin H2-induced aggregation in indomethacin-treated resuspended platelets but did not inhibit thromboxane synthesis. We therefore conclude that 13-APA acts as a direct antagonist of the platelet thromboxane/endoperoxide receptor.


Quantitation of phosphatidylcholine secretion in lung slices and primary cultures of rat lung cells. Rat lung slices and isolated rat lung cells were used to study the secretion of phosphatidylcholine by the lung in vitro. The rate of incorporation of [(3)H]choline by lung slices was 20-fold greater than by liver slices and 4-fold greater in lung cells compared to confluent skin fibroblasts. Labeling lung slices or cells with [(3)H]choline for up to 8 hr failed to reveal a significant amount of labeled phosphatidylcholine in the medium of either system compared to the medium from liver slice or fibroblast controls. Labeling of isolated lung cells for up to 24 hr, with or without 10% fetal calf serum, also showed no significant difference in the amount of labeled phosphatidylcholine in the medium compared to control fibroblast cultures. Washing labeled lung slices or cells with a nonlysing concentration of Triton X-100 (0.05%) did not selectively release labeled phosphatidylcholine, indicating that any secreted phosphatidylcholine did not adhere to the surface of the lung slices or cells. Experiments were performed to determine whether the small amount of phosphatidylcholine in the medium and detergent-released phosphatidylcholine was similar to the tissue and cell phosphatidylcholine. The saturated fatty acid composition of the phosphatidylcholine released by Triton X-100 and in the medium (from lung slices) was identical to that of the tissue phosphatidylcholine. In addition, the relative labeling rates of the phospholipids released by Triton X-100 and in the medium (labeled with [(14)C]glycerol) were identical to those of the tissue and cell phospholipids. Based on these results, we conclude that phosphatidylcholine is not secreted by lung slices and lung cells in large amounts compared to controls. The implication of these data is that pulmonary surfactant material may actually not be secreted by the lung in vitro, and perhaps in vivo, in the manner that is currently generally accepted.


Release of platelet-derived growth factor from human platelets by arachidonic acid. Platelet alpha-granules contain a factor that stimulates the proliferation of arterial smooth muscle cells and may play a role in atherogenesis. We have studied the role of arachidonic acid in mediating the release of the platelet-derived growth factor (PDGF) from human platelets. PDGF was assayed by stimulating of [(3)H]thymidine incorporation into DNA of mouse 3T3 cells. Platelet aggregation and the release of platelet factor 4,beta-thromboglobulin, and serotonin were also studied. A biphasic response pattern was observed when gel-filtered platelets were incubated with arachidonate over the concentration range 0.01-0.4 mM. At low arachidonate levels (approximately 0.025-0.1 mM), specific concentration-dependent aggregation and release of PDGF and of the other components were observed. This effect was not seen with any of five other fatty acids tested and was suppressed by indomethacin (25 muM). At higher arachidonate concentrations (approximately 0.15-0.35 mM), a concentration-dependent turn-off of both aggregation and release occurred. At these concentrations the platelets remained functional, and no release of lactate dehydrogenase was observed. A similar biphasic pattern of arachidonate-induced aggregation and release was observed with platelet-rich plasma, over a similar range of arachidonate to albumin mole ratios. These studies demonstrate that PDGF and other alpha-granule constituents can be released from platelets specifically by arachidonate via an indomethacin-sensitive pathway, most probably involving the platelet cyclooxygenase and conversion of arachidonate to prostaglandin metabolities. The mechanisms responsible for the turn-off of the specific arachidonate-mediated responses at higher arachidonate concentrations remain to be defined.


Topological organization of proteins in an intracellular secretory organelle: the synaptic vesicle. Intact synaptic vesicles prepared from the electric organ of the marine elasmobranch Narcine brasiliensis have eight major polypeptides demonstrable on sodium dodecyl sulfate gels. Six of these copurify with the synaptic vesicles during isolation of vesicles by chromatography on CPG-3000 and, by this criterion, are specific to vesicles. The other two are either shared by many membrane or are contaminants. One of these proteins comigrates with actin. Three different approaches were used to determine which proteins were exposed on the external, cytoplasmic surface of the vesicle and which were internal. The first was susceptibility to the proteases trypsin, Streptomyces griseus protease, and Pronase; the second was labeling by the membrane-impermeable reagent diazotized [125I]iodosulfanilic acid; and the third was iodination catalyzed by lactoperoxidase. In general, the three approaches give the same result: six of the eight proteins are on the external, cytoplasmic surface and two are accessible only after the vesicles are lysed by freezing and thawing or by detergents. Five of the vesicle-specific proteins are external and one is internal. The actin-like protein is internal. Proteins involved in the interaction of vesicles with the presynaptic membrane during exocytosis might be expected to be vesicle specific and external.


Decussion geometrics in the goldfish nervous system: correlation with probability of survival. In the goldfish, the optic nerve decussation occurs without intermingling of fibers from the two eyes. In two-thirds of juvenile and adult specimens, the left optic nerve is dorsal at the midline. In about 60% of the specimens, the decussation of Mauthner's neuron also has a left-dorsal-to-right (L/R) configuration. Concordance for decussation geometry is greater than 80%, with smaller specimens accounting for a disproportionate number of discordant cases. In embryos and very young larvae, the L/R configuration occurs in slightly less than 50% of optic chiasmata and in slightly more than 50% of Mauthner's cell chiasmata, and there is no significant tendency toward concordance. However, larval specimens that survive 1 month after hatching are markedly skewed toward adult decussation patterns and somewhat skewed toward concordance. These observations, together with results of cannibalization and predation experiments, suggest that the adult population pattern can be achieved through selection pressures against discordant individuals and, perhaps separately, against individuals with right-dorsal-to-left (R/L) chiasmatic configurations. Decussation patterns correlate with embryonic postures, larval orientation preferences, and growth rates after hatching, which may themselves serve as a basis for selection.


Selection of variant neuroblastoma clones with missing or altered sodium channels. Neurotoxins that cause persistent activation of voltage-sensitive sodium channels are highly cytotoxic to electrically excitable neuroblastoma cells. These toxins were used as selective agents to isolate variant neuroblastoma clones with missing or altered sodium channels. Of ten resistant clones analyzed, seven lacked functional sodium channels and one had a specific 40-fold increase Kd for scorpion toxin and altered voltage dependence of scorpion toxin binding. The phenotypes of these cell clones were stable for more than 100 generations, indicating that they were the result of stable genetic change.


The Bezold reflex: a special case of the left ventricular mechanoreceptor reflex. Our previous finding that increasing myocardial contractility caused reflex systemic hypotension, the left ventricular (LV) mechanoreceptor reflex, suggested that the classical Bezold reflex (systemic hypotension and bradycardia after intracoronary administration of veratrum alkaloids) may be initiated by these same LV mechanoreceptors. In our working LV preparation with the coronary and systemic circulations isolated and perfused separately, intracoronary injection of veratrum alkaloids, like that of catecholamines or ouabain, had a positive inotropic effect which produced the hypotensive response typical of the LV mechanoreceptor reflex. To test directly if veratridine's positive inotropic effect initiates the Bezold reflex, verapamil, which blocks the slow Ca(2+) channels of myocardial cells but leaves intracardiac nerves unaffected, was injected by the intracoronary route to prevent the increased contractility from intracoronary injection of veratridine which also abolished the reflex hypotension, demonstrating conclusively that increasing myocardial contractility and thereby activating LV mechanoreceptors but not chemoreceptors initiates the Bezold reflex. Contrariwise, decreasing contractility or cardiac asystole by administration of tetrodotoxin, verapamil, or EDTA resulted in an increase in the systemic resistance, indicating that changes in the magnitude of the stimulus initiating the LV mechanoreceptor reflex (i.e., changes in myocardial contractility) lead to directionally opposite changes in peripheral resistance, as in the sino-aortic mechanoreflexes. Thus, it is concluded that the Bezold reflex is a special case of the LV mechanoreceptor reflex. The latter, by means of feedback mechanisms, functions normally by continuously matching the peripheral resistance to the LV contractile state so as to maintain the arterial pressure constant, thereby playing an important role in blood pressure regulation.


Prostaglandin A1 induces differentiation in Friend erythroleukemia cells. The effect of different prostaglandins and prostaglandin-metabolites on the growth and differentiation of Friend erythroleukemia cells (FLC) was evaluated. The prostaglandin-metabolites, thromboxane B2 and 6-keto PGF1 alpha, were completely inactive, while PGE1 inhibited slightly and PGF2 alpha stimulated the replication of FLC. PGA1 was found to be the most active compound. It profoundly inhibited the replication of both DMSO-treated and undifferentiated FLC. Most importantly, PGA1 alone induced differentiation in FLC, stimulating hemoglobin production over a five-day period. PGA1-stimulated differentiation was completely suppressed by the addition of 10(-6)M hydrocortisone. Finally, treatment of DMSO-differentiated cells with PGA1 (but no DMSO) prevented the return to the undifferentiated state.


[Dental amalgam corrosion].

[Dental amalgam corrosion]. Corrosion of amalgam is a natural phenomenon of the behaviour of a metallic substance in the oral environment. Corrosion is a process in which we have a anodic reaction resulting in a loss of electrons and a cathodic reaction resulting in a captation of electrons. One single amalgam restoration in a electrolitic environment as the saliva shows areas acting as anode and others acting as cathode. Certain factors may influence the corrosion: they are the composition of the alloy, the particle size and form, the mercury content, the presence of more or less important quantity of gamma-2 phase, the degree of surface finishing, etc. It is likely that the dispersion phase alloy are more resistant to corrosion than the conventional alloys and that fluoridates amalgams are also more prone to corrosion.


[Buccal lupus lesions (author's transl)]. Lupus is a particularly polymorphic disease involving all medical disciplines and inciting the publication of an increasing number of papers. The two clinical types of lupus (acute disseminated lupus erythematosus and chronic lupus) have, until recently, led to doubts as to the unified nature of the disease. Grouping the symptoms and the results of their investigation in a computer has shown that there are 14 elements which occur most frequently, though there is no absolute pathognomonic sign of the disease. Oropharyngeal ulcerations, without being typical, are on of the fundamental signs and were found in 10% of cases of chronic lupus. The Gougerot-Sjögren syndrome is sometimes found, but it is not one of the fundamental signs : its existence, however, has enabled progress to be made during research into the auto-immune physiopathology of lupus disease.


[Cowden's disease. A report on two cases in Swiss families (author's transl)]. Described by Lloyd and Dennis in 1963, Cowden's disease is a familial affection in which are found associated lesions of the buccopharyngeal mucosa (papillomatosis, papules, scrotal tongue), various types of skin lesions (verruciform keratotic papules, papilloma, lipoma, angioma, etc...), visceral manifestations usually related to lesions in the thyroid, breasts, or digestive tract (hamartoma or malignant tumors), and skeletal dysmorphism. The two patients were from different families, were aged 65 and 28 years, and had presented for consultation because of buccal mucous membrane papillomatosis, and in one case, hemorrhagic gingivitis. Cowden's disease was diagnosed from the clinical examination, past history, and investigations after hospital admission. The main value of the mucosal and cutaneous lesions is their diagnostic significance, enabling a synthesis of the multiple dispersed manifestations to be made, and a possible visceral tumor to be discovered in the early stages.


[Cowden's disease : multiple hamartomata syndrome (author's transl)]. The third case of Cowden's disease to be discovered in France is reported. The authors emphasize the means for the diagnosis of multiple hamartomata, the typical symptoms and signs in the case reported, and that the family history included a daughter aged 8 years with macrocephaly, and slight mental retardation, who had already been operated upon twice for tonsillectomy.


[Kaposi's disease (author's transl)].

[Kaposi's disease (author's transl)]. A case of Kaposi's disease is reported with a lesion in the buccal mucosa, and the history, classification, signs and symptoms, and treatment of this disease are described.


[Ulcerovegetating syphilid of the buccal mucosa (author's transl)]. The authors report a case of ulcerovegetating lesions of the buccal mucosa in a patient with confirmed cutaneous and biological secondary syphilis, and discuss the diagnostic features of ulcerovegetating mucous syphilids which correspond to the lesion originally described by Milian, and to the diagram illustrating the work by Grinspan quoted in the references.


[Gonococcal stomatitis or polymorphic erythema associated with a gonococcal urethritis (author's transl)]. A case of polymorphic erythema which could have been caused by a gonococcal infection is used as a basis for retracing the evolution of the gonococcus at the present time, describing the conditions necessary for its diagnosis, and discussing the possible interpretation of this case.


[Buccal metastases from soft-tissue cancers. A report on 3 cases (author's transl)]. Histological examination of the gingivovestibular lesion in the first case with known kidney cancer, confirmed the metastatic nature of the buccal tumor and its origin. The second case raised the problem of the diagnosis of a labial swelling occurring during widespread metastases from breast cancer. The third case was more complex as the palatine lesion was seen initially, and metastatic seeding extremely rapid. This led to the hypothesis only, that bronchial cancer was the primary lesion.


[Reconstruction of the frontal bone after trauma (author's transl)]. Reconstruction of the frontal bone after trauma has to be performed in a different manner according to whether the frontofacial lesions are associated or not with neurosurgical disturbances. The lesions in the first case mainly affect all the orbital borders and the anterior walls of the frontal sinuses. Reconstruction of the frontal bone usually requires osteosynthesis to the metallic threads of the various fragments applied, but the treatment of the frontal sinus region must be modulated in relation to the possible existence of posterior wall lesions. Frontofacial lesions associated with neurosurgical disorders require the combined efforts of a neurosurgical and maxillofacial team at the same time. The frontal sinus extends into the cranial region in all cases. Frontal bone reconstruction requires the use of various techniques (osteosynthesis to metallic threads, grafts taken from the iliac or other bones), according to the site and severity of the traumatic lesion.


[Hour-glass craniofacial teratoma (author's transl)]. A male infant was born with a voluminous left jugal swelling identified by biopsy on the 8th day as a mature benign teratoma. Further investigations were conducted because of mild exophthalmia, including an electro-encephalogram and craniofacial computer tomography. An enormous frontotemporal intracranial extension of the tumor was discovered on the same side. The tumor was completely excised at the age of 1 month by a two-stage operation : neurosurgical followed by maxillofacial eight days later. The postoperative course was uneventful. There was no relapse after one year but because of the consequences of intracranial excision the prognosis was very poor. The authors discuss this case and review those in the published literature in order to describe the diagnosis, treatment, and prognosis of this extremely rare craniofacial teratoma of the newborn.


[Value of zonography in parotid sialography (author's transl)]. The authors describe a radiological technique for improved interpretation of parotid gland sialography, using thick section tomography (zonography).


Rough endoplasmic reticulum-associated tubular structures occurring in acute promyelocytic leukaemia. Intracytoplasmic tubular structures (TS) which have not yet been described were found electron microscopically in a large percentage (26.5%) of leukaemic cells from a 25-year-old female with acute promyelocytic leukaemia. The TS were bundles of tubules measuring about 90 nm in diameter and continuous with rough endoplasmic reticulum, the maximum of the TS attaining 3.0 micron in length and 0.53 micron in width. The TS were mostly solitary, occasionally 2 or 3 within a cell in a plane of sectioning and were usually located around the Golgi region. By three-dimensional observation with a goniometer stage, it was confirmed that the TS showed latticework of rings in cross sections and fascicular tubular arrays in longitudinal sections. In the latter, some tubules exhibited meshworks of polygonal vesicles measuring about 20 nm and cross striations of filamentous bristles with the periodicity of about 22 nm. The origin and nature of the TS are discussed.


Periodic acid-Schiff reaction changes during the course of relapsing lymphoblastic leukaemia. Bone marrow smears taken at diagnosis and subsequent haematological relapse from 41 patients with lymphoblastic leukaemia (ALL) were compared to assess the proportion of blast cells showing a positive periodic acid-Schiff (PAS) reaction at different stages of the same disease. 20 patients never displayed more than 10% PAS positive blasts at any stage, and of these 8 never had any PAS positivity at all. 7 patients maintained a proportion of PAS positive cells over 10% of the total but with no loss or gain in the numbers of such cells. 14 patients showed a significant change in the PAS reaction during the course of their disease, 7 showing a marked reduction in PAS positive cells, and 7 showing an equally marked increase. From these results it seems that the PAS reaction does not identify a sub-type of ALL or mark the degree of differentiation of a given case. It could be indicative of a specific stage of the lymphoblast mitotic cycle.


Fanconi's anaemia developing erythroleukaemia. A 15-year-old boy with Fanconi's anaemia (FA) for 10 years developed acute erythroleukaemia. During the leukaemic phase, granulopoietic stem cells (CFUc) were absent from his bone marrow and blood, but proliferation and differentiation of bone marrow cells could be seen in semipermeable diffusion chambers in vivo, and globin synthesis of erythroblasts had become imbalanced. Chromosomal lesions of peripheral blood lymphocytes differed in the leukaemic phase from those in the pancytopenic phase. These data indicate that erythro-, myelo-, thrombo-, and lymphocytic cell lines all were involved in both the leukaemic and the pancytopenic process. It is suggested that terminal myeloproliferative disease developed as part of the natural history of FA.


Acute leukaemia and Gaucher's disease.

Acute leukaemia and Gaucher's disease. As association between Gaucher's disease and haematopoietic neoplasias has been reported in only a few instances. The authors recently encountered 2 cases of adult Gaucher's disease that developed acute myelogenous/myelomonocytic leukaemia. Both individuals had a pre-leukaemic phase. These cases are reported.


The effect of thioguanine on a combination of daunorubicine, cytarabine and prednisone in the treatment of acute leukaemia in adults. The Finnish Leukaemia Group. 194 adults with acute leukaemia were randomly allocated to be treated with a combination of daunorubicine, cytarabine and prednisone either with (RAP + T) or without (RAP) thioguanine. A remission was achieved in 37% of 101 patients treated with RAP and in 35% of 93 patients treated with RAP + T. The survival and length of remission were similar in both groups. Neither regimen was superior to the other in any type of leukaemia nor in any age group of patients. In 9 of the patients failing to remit with RAP treatment a remission was obtained with other chemotherapy, while none of the patients not responding to RAP + T achieved a remission with further chemotherapy.


Codominant translational mutants of Chinese hamster ovary cells selected with diphtheria toxin. Diphtheria toxin-resistance markers in two translational mutants, CH-RE1.22c, possessing no toxin-sensitive EF-2 (class IIa), and CH-RE1.32, with 50% toxin-sensitive and 50% toxin-resistant EF-2 (class IIb), behaved codominantly in somatic cell hybrids. There was no complementation in hybrids formed between the two resistant mutants. The mutant parents and their hybrids, except those formed by fusion of CH-RE1.32 and wild-type cells, grew in the presence of toxin. To explain these results we suggest that CHO-K1 cells possess two functional copies of the gene for EF-2 and that CH-RE1.22c and CH-RE1.32 represent the homozygous (R/R) and heterozygous (R/S) states of resistance at the EF-2 gene locus. The failure of hybrids formed between CH-RE1.32 and wild-type cells to grow in toxin is a gene dosage effect. Codominant class IIa translational resistance is a selectable marker for the isolation of hybrids. It can be combined with a second, recessive, marker to provide a cell which is a "universal hybridizer" (10).


Marsupial--mouse cell hybrids containing fragments of the marsupial X chromosome. Hybrids were obtained from fusions of HPRT-deficient mouse fibroblasts and marsupial lymphocytes. These hybrids retained no identifiable marsupial chromosomes, but all expressed the marsupial form of HPRT. Half the clones also expressed marsupial PGK-A, and half of these also marsupial G6PD; no other marsupial allozyme markers were detected. Since G6PD is known to be sex linked in these species, we conclude that Hpt and Pgk-A are also located on the X chromosome and the markers lie in the order Hpt-Pgk-A-Gpd.


Genetic analysis of tumorigenesis: IV. Chromosome reduction and marker segregation in progeny clones from Chinese hamster cell hybrids. Hybrid cells produced by the fusion of pairs of cells, one a tumorigenic derivative of CHEF/16 and the other a nontumorigenic derivative of CHEF/18, give rise to clones which are largely tetraploid, but rare reduced hybrids with chromosome counts in the diploid range have been recovered from tumors of hybrid origin. This paper describes the recovery in cell culture of reduced hybrids in the diploid range by selection with 5-bromodeoxyuridine (BrdU) or methylcellulose as well as by growth in culture of cells from excised tumors. All selected subclones were tumorigenic and resistant to BrdU, but they segregated for resistance to 6-thioguanine. Unselected subclones were tetraploid, nontumorigenic, and sensitive to both drugs. These data show that chromosome reassortment as well as extensive chromosome reduction both occur in a small fraction of the population during growth of each hybrid clone.


The influence of observer performance in radiographic caries diagnosis. Experienced oral radiologists were asked to report whether the proximal surfaces of extracted permanent premolars were radiographically positive or not with regard to carious lesions. The percentages of true positive and false positive diagnoses for each observer were plotted in a graph, displaying a reciprocal relationship. On the basis of differences between observers theoretical calculations were made of the relative number of carious lesions that would have been reported by different observers in populations differing with respect to caries prevalence. It was shown that differences in reported prevalences do not only depend on variations between observers but also upon underlying true prevalences. The implications with regard to epidemiological studies are discussed.


Radiographic caries diagnosis and treatment decisions. Data regarding the radiographical appearances of proximal permanent tooth surfaces with and without clinical cavities were collected from the dental literature. On the basis of these data, calculations were made of the probabilities that decisions to restore proximal surfaces coincided with the presence pf clinical cavities. Such calculations were made with respect to different prevalences of carious cavities as well as the extents of the radiographically registered lesions. In populations with low caries prevalences there is a considerable risk for overtreatment if all surfaces with radiographically registered lesions would be restoratively treated without consideration to the extent of the radiolucent area.


Ostectomy in the mandible.

Ostectomy in the mandible. Fourteen patients, diagnosed as cases of apertognathism, were subjected to ostectomy in the premolar or molar region on both sides. The operations were made with an intraoral approach in the premolar region anterior to the molar foramen and extraorally in the molar region. The mandibular segments were immobilized by cap-splints after the operations. A follow up, clinically and roentgenologically, was made after the splints had been removed and, in addition, one and five years later. The aesthetic as well as the functional results of the operations were very good. Only minor changes in the occlusion occurred during the follow up period. In a few patients minor unilateral decreases of sensibility were observed. The late results of the operations must therefore be regarded as good.


Prevalences of removable dentures and edentulousness in an adult Swedish population. In three geographic regions, urban, suburban and rural from the middle of Sweden 20,333 adults (over 15 yrs of age) were examined clinically regarding denture wearing and edentulousness. 30.3% had removable dentures of some kind and 1.0% were edentulous and did not wear any dentures. Removable dentures were significantly more common amongst females (33.8%) than amongst males (26.8%). For complete dentures in both jaws this sex differences was even more pronounced, 19.3% and 11.4%, respectively. The prevalence for removable dentures was highest in the rural region, and lowest in the suburban. This finding may, however, be partly explained by the different age distribution within the areas examined.


HLA-B17 and the HLA-A1, B17 haplotype in acute myelogenous leukemia. Seventy-nine Caucasians with acute myelogenous leukemia (AML) were genotyped to determine whether AML, the induction of remission or patient survival were associated with particular HLA phenotypes or haplotypes. HLA-B17 and B27 were increased in AML patients over 40 years of age. Combined analysis of four independent studies indicates that HLA-B17 is significantly but weakly associated with AML, relative risk = 1.48 (.01 less than P less than .025). The A1, B17 and Aw24, Bw35 haplotypes occurred more frequently in the AML group as compared to racial and geographic controls (uncorrected P = 0.0068 and 0.0098, respectively Fisher's Exact Test). Induction of remission occurred less frequently in patients with the B17 phenotype as compared to patients lacking this antigen (P = 0.047). Patient survival was associated with remission status (P = 0.002) but was not significantly associated with particular HLA phenotypes or haplotypes. These results indicate that a gene or genes in the HLA-B region of the major histocompatibility complex can influence susceptibility to AML and also the response to chemotherapy.


Shortened platelet survival in acute promyelocytic leukemia. A platelet kinetic study in 2 patients with acute promyelocytic leukemia (APL) associated with disseminated intravascular coagulation (DIC) demonstrated that the platelet survival was characteristically shortened. These findings suggest increased platelet consumption during the process of DIC in APL.


[A new method in plastic surgery for the correction of the lip red in bilateral labial clefts]. Having assessed the correcting operation according to Kapetansky and a modification according to Juri and co-workers, the author presents his own method designated by the term "butterfly plasty".


[1st animal experimental results with Bio-Vitroceramic implantations]. In the framework of an expensive research project, the author made animal experiments to compare the bio-active behaviour of bio-vitroceramic implants with that of implants currently used. After experiments for more than a year, the author presents his first scanning electron microscopical findings that are indicative of direct binding of the implant to the bone.


[Therapy of the styloid syndrome].

[Therapy of the styloid syndrome]. Resection yields the best results in treating pain-producing megastyloids. With due regard to their courses, the extra-oral approach may be used for exposing them, but the intra-oral approach is very often more indicated. Due to the immediate proximity of the great cervical vessels, the resection in the high region, which is most likely to avoid failures, is not riskless. For this reason, the author uses for the intervention an instrumentarium developed on the basis of his own suggestions.


[Problems in the admission to in-hospital oral surgical care from the patient's viewpoint--results of patient interviews in the hospital for dental and maxillo-facial surgery of the Karl Marx University, Leipzig]. The author deals with the results from the interviewing of oral surgery patients on their problems concerning the sending and the admission to the hospital, with special attention to the problems of waiting for admission, the familiarization with the clinical environment and the improvement suggestions of the patients. The conclusions concern tasks arising from the medical and dental care for inpatients as well as for outpatients.


[Results of an analysis of waiting time at a maxillofacial surgery outpatient department of a university hospital]. Waiting periods in public health institutions not only stress the patients and the personnel, but they are also attended by considerable economic disadvantages. For this reason, the waiting periods of patients of a maxillofacial outpatient department in the various examination and treatment stations were analyzed with the help of accompanying leaflets. According to the results obtained, the sequence of examinations and treatments was verified and reorganized with regard to the three main processes: research, medical care, and education and training. The control by a second analysis evidenced a reduction of the total waiting period from 46.0 to 36.9 minutes per patient. Further possibilities of reducing the waiting periods are indicated, and reasons are given for the necessity for scientific work organization in public health institutions.


[Further results from the Berlin morbidity study 1972. 2. Educational level and incidence of periodontal diseases]. The ratio of subjects without periodontal diseases to those with periodontal diseases is 40:60 for individuals having gained a primary school-leaving certificate, and 53:47 for individuals with higher educational level.


[Graduate teaching aspects to be considered in the planning of undergraduate dental courses]. The general part of the present paper deals with questions concerning the demanding conceptual design of teaching from the aspect of creative studying. The students' qualification for responsible scientific work and creative studying is considered to be an essential question of the conceptual design of the demanding process of learning and acquiring the scientific bases. Thus, it will be possible to improve effectively the attitude towards studying and the engagement in creative achievements (in a narrow sense or in explicit forms of organization). In the empirical part of the present paper, the results from the interviewing of 240 dental students on relevant conceptual factors of teaching are discussed. An analysis of situation led to valuable insights and conclusions as to more effective teaching in the framework of stomatological study.


[Dental caries and pregnancy].

[Dental caries and pregnancy]. In a Berlin population comprising 588 women aged between 18 to 50 years, no increase of the DMF index was observed in multiparae. Nevertheless, complete dental treatment was stated in them less often.


[How current is the rubber dam method?]. The coffer dam application is still a condition necessary for the successful treatment of pulpitis and periodontitis. It keeps the operating area perfectly dry and prevents incidents in endodontics. The various coffer dam techniques and their indications are described.


Prognostic factors and complications of vitreous surgery. Vitrectomy has provided a means for the restoration or amelioration of drastically reduced visual acuity in selected patients. Many patients have, as a consequence of the deleterious natural history of their disease, suffered permanent impairment of vision, but in some cases even a relative therapeutic success is very gratifying. Nonetheless, in its present state, vitreous surgery is not without a significant incidence of complications which are enumerated in this paper.


Closed vitrectomy surgery using modified Grieshaber apparatus. A closed vitrectomy procedure was carried out on 32 eyes at the Manchester Royal Eye Hospital between October, 1976, and February, 1978. The Grieshaber/Boston Retinal Foundation Vitreous Cutter was used exclusively for these operations. Details of the simplified surgical technique are described and illustrated. The results are assessed from the technical aspect (the achievement of a clear light path to the retina) and from the functional aspect (the postoperative visual acuity). The technical success was of the order of 80 per cent.


Anterior segment applications of vitrectomy techniques. We have used vitrectomy instrumentation and techniques to treat a variety of conditions affecting the anterior segment, including: (1) Various conditions causing an occluded or inadequate pupillary space, (2) Complications of vitreous gel in the anterior chamber, (3) Growth of epithelium into the anterior chamber, (4) Selected indications for lens surgery, (5) Flat anterior chamber, (6) Total hyphaema. The vitrectomy methods provided improved capabilities, including controlled excision of intraocular tissues under optimum visualization using a closed-eye system with normal intraocular pressure and the ability to use accessory techniques such as intraocular diathermy. Although new difficulties are introduced by reliance on complex instrumentation and possible complications from excessive traction or use of large volumes of intraocular irrigating solution, our experience in using these techniques to treat selected anterior segment conditions has been, favourable.


Glaucoma implants. Their use in difficult cases of glaucoma. Implants with a regime of fibrosis suppression (prednisolone 30 mg daily plus fluphenamic acid 200 mg three times a day for 6 to 8 weeks) controlled the intraocular pressure (applanation tonometry (AT) less than 21 mm Hg) in seven out of ten patients with different types of glaucoma. One implant was removed after erosion of the conjunctiva and sclera; there were no other serious complications. Use of the implant is indicated after failed drainage operations or as a primary procedure in glaucoma associated with aphakia, chronic uveitis, vitreous in the anterior chamber, and scarring of the conjunctiva.


Field defect due to posterior cortical paraxial lens opacity. A case report. A 60-year-old man had variable visual acuity especially in the right eye for 2 years. The right field of vision showed a hemianopic defect on the nasal side (in addition to reduced visual acuity which was due to nuclear cataract). A localized posterior cortical cataract on the temporal side of the right lens accounted for the nasal field defect since the defect disappeared almost completely on dilatation of the pupil, and completely after cataract extraction. Asymmetrical opacities in the optical media far enough anterior or posterior to the plane of the pupil will produce asymmetrical field defects. (An opacity close to the plane of the pupil will produce concentric constriction.)


Optochiasmal arachnoiditis after rupture of an anterior communicating artery aneurysm. This paper reports two cases of optochiasmal arachnoiditis which followed subarachnoid haemorrhage. In one case the visual deterioration was arrested and improved by the administration of systemic steroids.


Effects of ultrasonication of the rabbit lens in situ as evaluated by analysis of crystallin composition. Ultrasonication and aspiration is a recently developed method of lens extraction. We have attempted todetermine whether extraction by this procedure is total or partial and, if so, whether certain regions or components are preferentially removed, and also whether the contents of the lens are physico-chemically modified by the process of extraction. The water-soluble and urea-soluble protein fractions from the ultrasonicated lenses and the contra-lateral intact lenses of six rabbits were analysed by isoelectric focusing. Differences are found in the crystallin content which indicate that extraction is frequently non-representative and that beta- and gamma- crystallins are modified by the process.


Disseminated intravascular coagulopathy. Disseminated intravascular coagulation (DIC) is not a disease but a pathological process with widespread thrombus formation in small vessels; it occurs in many systemic conditions that stimulate the intravascular clotting mechanism. There may be widespread tissue involvement, and any tissue in the body may be affected, especially in the kidney, brain, liver, heart, and lungs. This abnormal coagulation is now commonly referred to as disseminated intravascular coagulopathy. It is prone to occur in obstetrical complications, in cancer, after transplantations, and where there has been tissue damage, such as burning, crushing, and surgery, all of which release thromboplastin into the circulation. It may also occur in Gram negative bacterial systemic infections, in antigen-antibody reactions, and in thrombotic thrombocytopenic purpura. When the eye is involved, the thrombi occur in the choriocapillaris, and are usually limited to the submacular and peripapillary choroid. The anterior parts of the eye generally escape involvement. Visual symptoms may be early, and may be due to central choroidopathy or to secondary retinal detachment.


[Polymorphism of a tumor cell population and selective processes. III. A change in the correlation of tumor cell subpopulations of the ascitic strain of Ehrlich-I.Ch.Ph. under the influence of glucose and sodium succinate]. A study was made of the action of glucose or sodium succinate on subpopulations of the Ehrlich-I.Ch.Ph. ascite strain characterised by markers "A1" and "A", resp. After i.p. injection of glucose the amount of "A1"-cell reached 50 and almost 100% on the 5th and 7th day of tumor growth. After the transplantation of "A1"-cells into intact animals, a homogenous cytogenetic feature of subpopulation persisted during 2 passages only. Kinetics studies of a subvariance of the Ehrlich-I.Ch.Ph. tumor containing "A1"-cells show that the tumor growth rate and grade of malignancy slightly differ from those seen in the controls.


Giant-cell pneumonia and lung tumorlets in a leukemic four-year-old child. The authors report a case of a 4-year-old child whose death resulted from acute lymphoblastic leukemia (ALL) and giant-cell pneumonia. Typical lung tumorlets were occasionally observed. On electron microscopy study it was possible to demonstrate a large number of neurosecretory granules in the tumorlet's cells, identical to those present in Kulchitsky's cells. These studies suggest the occurrence of an abnormal immunitary process.


[Glucose-6-phosphate dehydrogenase and pyruvate kinase activity and the content of adenosine triphosphate in the erythrocytes of leukemic patients]. The activity of G-6PD and PK in erythrocytes and ATP content in erythrocyte hemolysate in patients with leucosis (acute, chronic myelo- and chronic lympholeucosis) was investigated. A reduced G-6PD activity was found in the majority of the patients examined, being most markedly manifested in the patients with chronic lympholeucosis and least--in the patients with chronic myeloleucosis. About 50 per cent of the patients with acute leucosis and chronic lympholeucosis have reduced PK activity and in those with chronic myeloleucosis -- increased. ATP content is increased in patients with chronic myeloleucosis and in the rest of the patients -- normal. A disturbance of glucose metabolism in erythrocytes in patients with leucosis is concluded to exist.


Initiation in Papua New Guinea: psychohygienic and ethnopsychiatric aspects. Initiation ceremonials in traditional Papua New Guinea and North American Indian cultures serve important psychohygienic functions in establishing the youth's final identity and thereby warding off the frustration, anxiety, and depression which are associated with anomie and role confusion. Inititations in Papua New Guinea are presented as a process of social learning in which group consciousness and loyalty are established through revelation of ancestral secrets, testing by ordeals, and ego-stregthening rewards. Structually patterned archetypal collective symbols gain direct access to the young person's unconscious when skillfully transmitted in the initiatory psychodrama of death and rebirth. Medical complications occuring during initiation procedures are rare accidents which have to be weighed against the psychological and social benefits for individual and group. Initiation ceremonials help the young to achieve a sense of sexual and socio-cultural identity from which feelings of emotional security and social belonging are derived.


Properdin factor B-polymorphism. An indication for the existence of a Bf O-allele. The polymorphism of the properdin factor B (Bf, C3-proactivator, GBG = glycin-rich-beta-glycoprotein) has been investigated by high voltage agarose gel immunofixation electrophoresis in 1115 unrelated persons from Southern Germany. Seven phenotypes were observed; the allele frequencies were calculated as BfS = 0.8094, BfF = 0.1790, BfSI = 0.0094, BfFI = 0.0022. A study of 94 parents with 98 children and 420 mother-child combinations showed no deviation from the assumed autosomal codominant mode of inheritance. In one additional family the findings suggested the existence of a silent allele at the Bf-locus.


[Pathogenesis of hyperkinesis in children]. Pheylalanine metabolism was studied in 56 children with various forms of hyperkinesias. It was found that in the development of slow and fast hyperkinesias a certain role belongs to dihydroxyphenylalanine (DOPA). It is probable, that in patients with Turett's syndrome the synthesis of DOPA is increased while in patients the excretion of phenylacetylglutamine was found to be disturbed: it was decreased in the patients with the fast and increased in the children with the slow hyperkinesias. Phenylalanine load led to a lowering of the DOPA level in the patients with Turett's syndrome; an intensification of the synthesis of phenylacetylglutamine and diminution of the intensity of hyperkinesias. L-glutamine load resulted in detoxication of the toxic phenylalanine metabolites which inhibited the DOPA synthesis, as well as in a short-time increase in the phenylacetylglutamine excretion and a moderation of tonic hyperkinesias that manifested by athetosis and dystonia. All this points out that in the development of various forms of hyperkinesias a certain role belongs to amino acid metabolites.


The extent of muscle tissue damage following missile trauma one, six and twelve hours after the infliction of trauma, studied by the current method of debridement. Spherical missiles with velocities around 1000 m/s were used to inflict missile trauma to one thigh of pigs. The determination of the impact and exit velocities made it possible to calculate the extent of energy transferred to the tissue. About one third of the pigs were treated surgically within one hour after the trauma, one third after a delay of 6 hours and one third after 12 hours. The surgical procedure consisted of debriding muscle tissue which showed impaired contractility and/or consistency and discoloration and/or lack of capillary bleeding - the current criteria of non-viability. The amount of debrided muscle tissue from each wound was weighed and the amount of debrided muscle tissue per joule transferred to the tissue was calculated. As compared to operations performed within one hour the average of the debrided muscle tissue per joule was greater after 6 hours delay and even still greater after 12 hours. However, the difference between the averages after 6 and 12 hours were not statistically significant.


Experimental soft tissue wounds caused by standard military rifles. Soft tissue wounds were caused by penetration of a bullet through both thighs in 42 pigs, fired from N48, M70, M16A1 and FAL rifles at 50 meter range. Four comparable groups of wounds, consisting mainly of injured muscle, were examined. Primary debridement was carried out 5-6 hours following wounding, multiple muscle biopsies up to 40 mm sidewise of permanent wound channel were taken for HE and histochemistry. Primary contamination was traced using marker aerobic bacteria. Bullet velocities at impact and exit were measured and transfer of energy to liver tissue calculated when feasible. The condition of the bullet after exit was precised. The surgical estimation of tissue injury, the mass of excised tissue and microsections have served to evaluate the extent of tissue injury. For 22.5 cm mean channel length the most injury was caused by the FAL, followed by M16, M70 and M48 rifles in declining order. Local effects are ascribed to bullet tumbling, break-up and energy transfer of corresponding missiles. Correlation of surgical and pathology methods is discussed.


The general and local response to injury related to complement activation. Injury can activate the complement system as well in vivo as in vitro. This activation might contribute to some of the phenomena seen after large injuries. Such general effects as fall in blood pressure, impaired ability to oxygenate blood and local effects such as accumulation of granulocytes and oedema formation might be influenced by complement activity. The in vitro experiments indicated an alternative pathway activation of whole complement by injured tissue or endotoxin.


Activation of the kallikrein-kinin system in bullet wounding. Anesthetized pigs were wounded in the thigh with a spherical missile at an impact velocity of 1500 m/sec. Arterial blood samples were taken before injury and at frequent intervals up to 12 hours after injury. Unbound (free) and total kallikrein (kallikrein + prekallikrein) were analyzed with a chromogenic substrate (S-2302, Kabi, Sweden) in which enzyme activity is determined spectrophotometrically by the development of a yellow colour. The results indicate an activation of the kallikrein-kinin system in bullet wounding, the significance of which remains to be determined.


Hemodilution and oxygen transport to tissue in shock. Shock was induced in sixty dogs by laparotomy, splenectomy and exteriorization of the small intestine for three hours. Resuscitation following different plasma substitutes was assessed by measuring hemodynamic parameters (mean arterial blood pressure, plasma volume, oxygen consumption, cardiac output and hematocrit). To justify a comparison, the colloids (albumin, ACD-plasma, dextran 40, dextran 70, gelatine) were given at the same concentration and dose (3.5 % solutions in saline at a dose of 1.5 g/kg or 43 ml/kg). Ringer's acetate was given in a three times larger volume (129 ml/kg). After reposition of the intestine, the infusion was given over a 20 minute period. During shock hemoconcentration developed, increasing from 50 to 59 % in hematocrit; mean arterial blood pressure dropped from 144 to 95 mm Hg; plasma volume decreased to 67 %; total body oxygen consumption decreased to 59 % and cardiac output to 21 % of initial values. Reposition of the intestine did not in itself improve these values. Ringer's acetate was effective up to about 30 minutes after infusion but did not differ from no infusion at four hours. Gelatine and ACD-Plasma had a similar effect up to two hours. Albumin, dextran 40 and dextran 70 increased and maintained plasma volume and oxygen consumption at preexperimental levels for up to at least four hours. It is concluded that a single infusion of albumin, dextran 40 or dextran 70 is superior to ACD-plasma gelatine or Ringer's acetate in restoring and maintaining oxygen consumption and hemodynamic functions, when given in comparable doses and volumes.


Effect of dextran infusion on the phagocytic and metabolic functions of the reticuloendothelial system in man. The reticuloendothelial system (RES) is part of the host defence and consequently plays an important role in shock and trauma, conditions often requiring volume replacement. Experimental studies have shown that the functions of the RES are hampered by plasma substitutes such as gelatin, starch and dextran. In this study six patients undergoing elective cholecystectomy were given 500 ml dextran just before the operation. At the end of the operation the phagocytic and metabolic functions were tested with microaggregated human serum albumin labelled with I125. The results indicate that there is a significant impairment of the phagocytic activitity of the RES in patients given dextran compared to controls not given dextran but saline. The metabolic activity was not significantly affected by dextran, however, suggesting that moderate dosage of dextran is not of great disadvantage to patients undergoing major surgery.


Human granulocyte function during dextran infusion. The granulocyte function was determined in healthy persons before and immediately after the infusion of 500 ml of dextran for 4 hours. The nitroblue tetrazolium (NBT) reduction reflecting the metabolic activity of these cells was measured both in cells at rest and after stimulation with E. coli, as well as with and without the addition of plasma. In 13 persons, the NBT-reduction was slightly but significantly increased in all four types of experiments after the dextran infusion. When dextran was added in vitro to granulocytes from normal volunteers, the NBT reduction of these cells was increased and the effect was dose-dependent. A surface effect of dextran together with phagocytosis of dextran molecules are the most probable reasons for the elevated NBT-reduction. - In 9 persons tested, the bactericidal capacity was not significantly influenced by dextran infusion. - When dextran was added in vitro to granulocytes neither the bactericidal capacity nor the chemotactic and random migration of the cells were changed. -


A gelatin energy methodology for estimating vulnerability of personnel to military rifle systems. Since 1965, estimates of bullet lethality have been obtained by firing the bullet of interest into a gelatin tissue simulant, measuring the energy deposited in the simulant and then relating the kinetic energy deposited to some previously determined empirical relationship between energy and P (I/H), the probability of incapacitation an infantry soldier, given a hit. Although the methods for recording the energy in the gelatin block have changed and the empirical relationships between P (I/H) and energy have undergone varying changes in sophistication and complexity, kinetic energy still remains the measure for quantifying the destructive physical damage which a high velocity military rifle bullet can inflict in biological tissues. It is the purpose of this paper to review the several gelatin correlation methodologies promulgated over the two decades, and through analysis of results of a recently completed study which evaluated these several methodologies, compare the precision of measurement techniques and evaluate the sensitivity of lethality predictions of each of the analytical models to the energy input parameters.


Steel ball effect-investigation of shooting at blocks of soap. Standard commercial glycerin soap was penetrated using 6 mm steel spheres. The impact velocities (VU) varied from 240 to 1700 m/s. The velocity of sphere within the soap (v), resistance force (F), cavity in soap (Z), and the maximal cavity diameter (D) were evaluated. Within the range of impact velocities used, the following functions have been determined F = a v2 X = b Vun, and D = c + d Vu Based on experimental evidence, the values a, b, c, d and n have been determined.


Break-up behaviour of some small calibre projectiles when penetrating a dense medium. A study of the break-up patterns of two types of standard ball small calibre bullets fired from a range of 100 meters against targets made of soft soap was performed. A flash X-ray set-up was used to study the behaviour of the bullets while penetrating the soap. A simplified calculation of the stresses affecting the bullets was performed. It turns out that one predominant factor in the break-up process is the tumbling of the projectile. The longitudinal rotation velocity of the bullets is often so great that this is sufficient to cause-centrifugal stresses of high enough magnitude to cause rupture of the bullet lead core. This in combination with the high retardation forces acting on a bullet penetrating a dense medium at large angles of yaw is sufficient to cause an almost inevitable break-up for fast-tumbling bullets.


Experimental studies on artificial ventilation using a tidal volume ventilator. Mechanics and dynamics of ventilation. In 24 piglets (2.7-24.5 kg b.w.), the mechanics of ventilation, the accuracy of dosage of respiratory volumes, and the influence of the ventilator's volume/pressure characteristics (Cvent, "internal compliance") on the dynamic course of insufflation were studied. A linear relationship was shown to exist between tidal volume and end-inspiratory tracheal pressure and between tidal volume and insufflation time. The insufflation time was reduced to about 50% of previously registered values. The error between set and registered tidal volume was found to be 6.0 +/- 2.7%. During the insufflation a linear relationship was found between the instant amount of delivered breathing gas and the corresponding endotracheal pressure change. The ventilator's Cvent did (and body size, total compliance and tidal volume did not) significantly influence the size of the direction coefficient for the linear instantaneous volume/pressure relationship, the magnitude of tracheal peak pressure and a short insufflation time, and vice versa. The use of greater power from the ventilator resulted in a significant shortening of the duration of insufflation and vice versa. The duration of insufflation is the parameter of choice in evaluating the efficiency of the ventilatory equipment. When the ventilator's performance is defined, measurements of the duration of insufflation may enable evaluation of conditions within the lungs.


The circulatory response to specific ventilatory patterns using a tidal volume ventilator. The circulatory response to different ventilatory patterns during artificial ventilation was examined in 17 sternotomized piglets. A constant CO2-tension level was maintained in all investigations by reference to analyses of the end-tidal infra-red CO2 fraction and arterial CO2-tension. The greatest variation in mean values for end-tidal CO2-tension was 0.2 kPa. Total compliance and lung compliance were lower at a ventilator volume/pressure quotient of 20 compared to those at 80 ml/kPa, and at f = 30 compared to f = 11 cycles/min. Higher cardiac output, lower pulmonary vascular resistance and systemic vascular resistance were measured at f = 11 (inspiration 20%) than at f = 30 (inspiration 50%). An increase in inspiration time by about 100% at the lower ventilatory frequency (f = 11) resulted in a significant but uncompensated decrease in cardiac output and stroke volume. These results demonstrate the value of a rapid insufflation in order to give longer expiration time per minute for the benefit of the venous return and cardiac output.