Quaternary structure of polynucleotide phosphorylase from Escherichia coli: evidence of a complex between two types of polypeptide chains. A new form of polynucleotide phosphorylase containing alpha and beta subunits was isolated (form B) by purification without preparative electrophoresis; this form was compared to the enzyme obtained by preparative electrophoresis purification (form A). The Stokes radius of these two forms are very different: 6.4 nm for form A and 8.7 - 9.0 nm for form B; on the other hand the sedimentation coefficients are close: 8.9 S and 9.9 S respectively. Such a result suggests that form B is very asymmetric. The apparent molecular weights, calculated from the Stokes radii and from the sedimentation coefficients, are approximately 365000 for form B and 252000 for form A. The latter is homogeneous on polyacrylamide gel, whereas the former yields two components, one of which behaves similarly to form A. Finally, whereas form A is composed of only one type of subunit, alpha, form B contains two types of chains: alpha (Mr 86000 +/- 5000) and beta (Mr 48000 +/- 2000) in stoichiometric proportions. From these results we believe that one should assume for form B the existence of a complex between form A and beta chains; the role of the latter still remains to be specified.


The modification of DNA-dependent RNA polymerase from Escherichia coli by an alkylating derivative of rifamycin SV. 3-(2-Bromo[1-14C]acetamidoethyl)-thio-rifamycin SV, abbreviated BrAcNEtS-Rif, and alkylating derivative of rifamycin SV was synthesized. A four-fold excess of BrAcNEtS-Rif inhibited the enzymic activity of RNA polymerase from Escherichia coli to 97%. Incubation of RNA polymerase with Br[14C]AcNEtS-Rif led to covalent substitution. The reaction of Br[14C]AcNEtS-Rif with enzyme at a ratio of 1.4:1 and a concentration of 63 nM was found to proceed with a half life of 1 h at 37 degrees C. The enzyme could be protected from reaction with BrAcNEtS-Rif by either rifampicin or the hybrid [poly(dT)]-[r(Ap)5a]. The modification of holoenzyme by Br[14C]AcNEtS-Rif in the presence of p-hydroxymercuribenzene sulfonic acid (pOH-HgBzSO3H) or 4 M LiCl occurred with faster kinetics and led to a higher degree of substitution. Reaction of Br[14C]AcNEtS-Rif with RNA polymerase core enzyme caused predominant substitution of subunit beta. In the case of RNA polymerase holenzyme the radioactive substituents were evenly distributed between subunits beta and sigma. Apparently the topology of the rifamycin binding site of holoenzyme, similarly to core enzyme, precludes attacks of nucleophilic functions from beta' and alpha, but it allows nucleophilic functions from subunits beta and sigma to react with equal probability on BrAcNEtS-Rif. In the presence of a 20-fold excess of pOH-HgBzSO3H, the modification of holoenzyme was drastically altered. Virtually all substitution took place on subunit beta', very little on beta and none on subunits sigma and alpha.


Role of ribosomal protein S1 in portein synthesis: effects of its addition to Bacillus stearothermophilus cell-free system. Products of the f2 phage RNA-directed protein synthesizing systems in vitro, derived from Escherichia coli and Bacillus stearothermophilus, were analyzed. In contrast to other reports, it was found that B. stearothermophilus ribosomes synthesized phage coat protein when the high-speed (100 000 X g) supernatant (S100) and ribosomal wash (crude initiation factors) of homologous origin were used. Furthermore, a marked stimulation of coat protein synthesis was observed with B. stearothermophilus ribosomes when either S100 or ribosomal wash or both from E. coli cells were used instead of the respective homologous components. The principle responsible for this stimulation was identified as the 30-S ribosomal protein S1 present in S100 and ribosomal wash. Purified S1 from E. coli at roughly one-to-one molar ratio to ribosomes resulted in about 10-fold stimulation of the incorporation of [14C]valine when added to the B. stearothermophilus system. This stimulation was observed mainly in the synthesis of coat protein and replicase.


Nucleotide sequences of Escherichia coli 16-S RNA associated with ribosomal proteins S7, S9, S10, S14 and S19. 1. A ribonucleoprotein fragment containing proteins S7, S9, S10, S14, and S19 was isolated in high yield from Escherichia coli 30-S ribosomal sub-particles. The same fragment was obtained whether ribosomes from E. coli strain A19 or MRE 600 were used, despite the fact that protein S7 differs widely between the two strains. RNA was extracted from this fragment and fractionated on gels containing 7 M urea, to reveal "hidden breaks". A well-defined and reporducible pattern of RNA fragments was obtained, with the main components being approximately 300, 240, 130, 115 and 75 nucleotides in length, respectively. The pattern of RNA fragments obtained was also independent of the strain of E. coli used. 2. Two-dimensional fingerprints were made from ribonuclease T1 hydrolysates of these RNA fragments, labelled with 32P, and the oligonucleotides were further analysed by digestion with either ribonuclease A or U2. The data obtained were fitted in detail to the new 16-S RNA sequence map of Ehresmann et al. (1975). Again no significant differences were observed between the RNA from E. coli A19 or MRE 600. The RNA sequences found lay in the region O'-D-E'-K-P-P'-E-A of the 16-S RNA, with a clear excision of several nucleotides in section E'-K. The total sequence length covered was approximately 430 nucleotides.


Cell-wall lipopolysaccharide from Escherichia coli B. The lipopolysaccharide of Escherichia coli BB and a number of R-phage selected (e.g. T3, T4) cell-wall-defective mutants were analyzed. From their lipopolysaccharides the respective core oligosaccharides were obtained. Following dephosphorylation, the oligosaccharides were methylated and analyzed by gas chromatography/mass spectrometry. This revealed the sugar sequence in the hexose-heptose region of the core. The linkage of heptose (Hep) to 2-keto-3-deoxyoctonate (KDO) was established as ... Hep 1,5 leads to KDO ... by methylation analysis. The substituted derivative of KDO was identified by gas chromatography and mass spectrometry. The KDO region contains three KDO units. Its structure was elaborated by (a) selective removal and identification of 7-phosphoryl ethanolamine-KDO (KDO-PN), (b) periodate oxidation and thiobarbituric acid reaction in conjunction with mild hydrolysis, (c) a modified methylation analysis. Phosphate substitution of E. coli BB core was studied by beta-elimination and using the information obtained with KDO-PN. The structures of the cell wall lipopolysaccharides from E. coli BB and cell-wall-defective mutants are given.


Malonyl and palmityl transferase-less mutants of the yeast fatty-acid-synthetase complex. 146 independently isolated mutants of the fatty acid synthetase gene locus fas 1 were subdivided into six different complementation groups. Three of these groups, Va, Vb and Vd, have not been described before. The mutant fatty acid synthetases isolated from representatives of complementation group Vb were specifically deficient in two component enzymes at the same time, the malonyl and palmityl transferases. Among more than 180 fas 1 and fas 2 mutants systematically screened for malonyl and palmityl transferase activities no mutant was found affected in only one of these two fatty acid synthetase component enzymes. From this it is concluded that both transfer reactions are catalyzed by the same enzyme. In any malonyl transferase-less fatty acid synthetase, neither of the two known malonyl binding sites, i.e. enzyme-bound pantetheine and the non-thiol binding site, accepts malonate. This indicates that malonate is transferred to both groups by the same enzyme. So far, no acetyl transferase-less fas mutants have been characterized. On the other hand, the mutants of two fas 1 complementation groups, Va and Vd, though negative in overall fatty acid synthetase activity had no deficiency in any of the known component enzymes which can be tested in vitro. A possible interrelationship between both findings is discussed.


Histidyl-tRNA synthetase from Salmonella typhimurium: specificity in the binding of histidine analogues. The topography of the active site of histidyl-tRNA synthetase has been investigated by determining Ki values for a variety of structural analogues of histidine, using the ATP-PPi exchange and tRNA aminoacylation reactions. Using these kinetic constants it has been possible to have a measure of the relative binding affinity of the enzyme for the histidine analogues. The following conclusions have been drawn: (a) the enzyme is stereospecific in the formation of aminoacyl-tRNA complexes, since the D-isomer of histidine does not influence the two reactions; (b) the carboxyl group is not required for binding; (c) bulky derivatives of the carboxyl group prevent the molecules from binding to the enzyme; (d) the amino group permits a good binding affinity; (e) the length of the ring side chain plays a very important role as point of attachment to the enzyme; (f) the kinds of heteroatoms on the ring determine the inhibitory properties of the analogues.


Biosynthesis of sterols by a yeast homogenate. Incorporation of mevalonic acid into cholesta-5,7,24-trien-3beta-ol and 5alpha-cholesta-7,24-dien-3beta-ol. Incubation of (3RS,2R)-[2-14C,2-3H]mevalonic acid and (3RS, 2S)-[2-14C,2-3H]mevalonic acid with mechanically disrupted yeast cells resulted in C27-metabolites. Two (14C5, 3H4)-metabolites, cholesta-5,7,24-trien-3beta-ol and 5 alpha-cholesta-7,24-dien-3beta-ol, were isolated and characterized. The impairment of the 24-methyl transferase system was confirmed by the lack of incorporation of 14C into the sterol fraction on incubation of S-adenosyl-L-[methyl-14C]methionine with the yeast homogenate. The results indicate that interference with the (C-24)-alkylating system did not prevent the transformation of lanosterol to the cholesta-5,7,24-trien-3beta-ol and to 5 alpha-cholesta-7,24-dien-3beta-ol. It can therefore be inferred that transformations of the nucleus and of the side chain can function independently. However our results do not provide a definition of the actual sequence of the metabolic events between lanosterol and ergosterol.


Determination of the complete amino-acid sequence of protein S4 from Escherichia coli ribosomes. After digestion of protein S4 with trypsin, all 32 tryptic peptides were isolated. Their amino acid compositions were analyzed and the sequence of the amino acids within the tryptic peptides was determined by means of a solid-phase peptide sequenator and by exopeptidases. Alignment of the tryptic peptides was established by analyzing and partially sequencing peptides isolated after digestion of the S4 protein with chymotrypsin, thermolysin and a glutamic-acid-specific protease. Further information about the alignment of peptides came from treatment of S4 with CNBr and with a lysine-modifying reagent.


The primary structure of tRNAIIArg from brewers' yeast. 1. Complete digestions with pancreatic and T1 ribonucleases. tRNAIIArg purified from bulk brewers' yeast tRNA by countercurrent distribution followed by two column-chromatographic steps was completely digested with pancreatic and T1 ribonucleases. Isolations of the products have been carried out either by column chromatography or by high-voltage electrophoresis. Analyses of the isolated nucleotides and olignoucleotides were in good agreement and indicate that this tRNA is composed of 76 nucltotide residues including 13 minor nucleotides. Overlaps resulting from the end-products of the two complementary digests led to a sequence of 25 residues. The primary structure of tRNAIIArg has been determined after partial digestion with T1 ribonuclease as described in the following paper.


Isolation and properties of a thymidylate-less mutant in Saccharomyces cerevisiae. A mutant, tmp3, has been isolated in Saccharomyces cerevisiae. Genetic and physiological analysis show that a single mendelian gene controls the multiple requirements for thymidylate, methionine, adenine and histidine and a neutral cytoplasmic petite character. Crude extracts of this mutant present a 60% decrease of serine transhydroxymethylase specific activity as compared to a wild-type strain.


Purification and properties of phosphoglucomutase from Fleischmann's yeast. 1. A procedure has been described for the purification of the major isozyme of yeast phosphoglucomutase of highest known specific activity. 2. The native enzyme has a molecular weight of about 65400 and was found to be homogeneous as judged by sucrose density gradient centrifugation, gel filtration, electrophoresis on acrylamide gel and ultracentrifugal analysis. In the presence of denaturing agents such as guanidine hydrochloride or sodium dodecyl sulfate, the enzyme dissociated into 32000-molecular-weight subunits. 3. As isolated, the enzyme has one mole of phosphate bound per mole of enzyme. Preparations incubated with 1.0 mM EDTA in 10 mM citrate buffer, pH 5.5 and dialysed against 10 mM metal-free citrate buffer, pH 5.5, contain no intrinsically bound Zn2+ and were enzymically inactive but fully active in the presence of 5 mM Mg2+ and 84% as active with 0.5 mM Zn2+. Simultaneous presence of both ions at these concentrations did not enhance activity. Enzyme was completely and irreversibly inactivated by preincubation with Be2+. Inactive enzyme had one mole of Be2+ bound per mole of enzyme. 4. Enzyme exhibited "ping-pong" kinetics rather than "random sequential". Km values for glucose 1-phosphate and for glucose 1,6-bisphosphate were calculated to be 2.34 times 10(-5) M and 2.24 times 10(-6) M, respectively. Rate of enzyme phosphate turnover was studied with rapid-mixing technique. The rates of 32P release from 32P-labeled enzyme and its appearance as glucose 6-[32P]phosphate were comparable and remained unaffected by addition of glucose 1,6-bisphosphate.


The thermodynamics and kinetics of conformational changes in 5-S RNA from Escherichia coli. Solutions of native, renatured and denatured 5-S RNA from Escherichia coli were each found to revert to a mixture of the denatured and renatured forms when incubated at elevated temperatures in the absence of divalent cations. The equilibrium and rate constants for the renaturation and denaturation reactions were studied as a function of temperature and concentration of added NaCl. The enthalpy and entropy of denaturation and the activation energies were determined. The activation energies for the renaturation reaction were found to be almost independent of the salt concentration, whereas the other parameters increased with the salt concentration. The results are interpreted in terms of a partially unfolded intermediate whose concentration is suppressed by the addition of salt.


An inhibitory effect of thymidine on its own conversion to nucleotide in Escherichia coli. The cellular levels of thymidine nucleotide pool in thymine-requiring mutants of Escherichia coli were followed. The pool levels of dTDP, dTTP and compound-X reached maximal values in much lower concentration of thymidine than those in thymine. In higher concentrations of thymidine, an inhibitory effect on its own conversion to nucleotide was observed. The inhibited step was suggested to be the conversion of dTMP to dTDP.


The regulation of arginine biosynthesis in Saccharomyces cerevisiae. The specificity of argR- mutations and the general control of amino-acid biosynthesis. The regulation of arginine biosynthetic enzymes in yeast is subjected to a double control. One level of arginine enzyme synthesis is under the control of an apo-repressor, called ARGR. ARGR molecules control specifically the arginine pathway. A second level of control of arginine biosynthesis has been disclosed. It also controls tryptophan, histidine, lysine, isoleucine-valine and probably many more biosyntheses. The general mechanism is turned on in leaky mutants in any of the amino acid pathways mentioned above.


Pools of ribosomal proteins in Escherichia coli. Studies on the exchange of proteins between pools and ribosomes. A labelling technique in vivo has been introduced which allows the tritiation of cell components with high specific activity during growth in rich medium. By this technique the pool size of each protein can be measured directly in the supernatant from centrifugation at 150000 times g. A measurable pool was found for the proteins S1, S2, S10, L1, L4, L7, L8/9, L10, L12, L21, and L25. Experiments on migration of ribosomal proteins from the supernatant to ribosomes (i.e. association) and vice versa (dissociation) demonstrate a remarkable constancy in the composition of the ribosome. There is no significant difference between ribosomes engaged or not engaged in poly-(Phe) synthesis.


Fluorescent labelling of Escherichia coli ribosomal sulfhydryl groups. The reactivity of the sulfhydryl groups of Escherichia coli ribosome has been investigated using a fluorescent label. Under denaturing conditions, all the --SH groups can be titrated. In the native form of the ribosome, six and less than one labels can be respectively conjugated to the 30-S and 50-S subparticles without loss of activity for poly(U)-dependent polyphenylalanine synthesis. The most reactive thiol groups belong to proteins S1, S12, S18, S21. The binding of mRNA plus tRNA to the 70-S ribosomes affects the spectroscopic properties of the labels showing that conformational changes are induced by these interactions. Furthermore, the treatment of these complexes by the labelling agent demonstrates that the --SH group belonging to protein S1 is partially protected whereas other thiol groups located on the 50-S subparticle become reactive.


Amino-acid composition of the covalent rigid-layer lipoprotein in cell walls of Proteus mirabilis. Cell walls of Proteus mirabilis in the stationary phase of growth contain a lipoprotein in covalent linkage to peptidoglycan and probably also in free form in the outer membrane. The protein moiety of this lipoprotein is composed of about 50 amino acids and has an approximate molecular weight of 5500. The Proteus lipoprotein has glycine and phenylalanine as specific components which are not present in lipoproteins of other enteric bacteria. Treatment of the peptidoglycan-lipoprotein complex of Proteus with trypsin leaves lysine as the only lipoprotein amino acid attached to the peptidoglycan. This suggests that in P. mirabilis, as in Escherichia coli, the lipoprotein is linked to the peptidoglycan by its C-terminal lysine residue.


The interaction of fusidic acid with peptidyl-transfer-ribonucleic-acid - ribosome complexes. The inhibitory action of fusidic acid on peptide-chain elongation was studied with systems in vitro directed by either polyuridylic acid or endogenous messenger (Escherichia coli polysomes washed with 1 M NH4Cl) or R17 RNA, and supplemented with either crude or purified elongation factors. In all cases strong inhibition of synthesis required high concentrations of the antibiotic (approx. 1 mM), while a similar inhibition of the EF-G-plus-ribosome-dependent GTP hydrolysis required between 10 and 100 times less antibiotic. Since most of the GTP hydrolysis observed was presumably due to free ribosomes (without aminoacyl-tRNA or peptidyl-tRNA), fusidic acid seemed to interact far more easily with these ribosomes than with ribosomes engaged in peptide-chain elongation. The role of the GDP-EF-G-ribosome-fusidic acid complex in the inhibition of polypeptide synthesis was assessed by measuring formation of this complex on polysomes engaged in peptide-chain elongation. Using purified elongation factors the complex formed on only 25-35% of ribosomes, as measured either by retention of [3H]GDP or by hydrolysis of [3H, gamma-32P]GTP. In contrast, with crude factors (S 100 extract) it formed on more than 70% of ribosomes. The results are compatible with the postulated role of the complex in polypeptide synthesis inhibition (blockade of the ribosomal acceptor site and subsequent inhibition of aminoacyl-tRNA binding) and indicate that formation of the complex takes place by overriding the control that prevents interaction of EF-G when the donor site is occupied by peptidyl-tRNA. In the polyuridylic-acid-directed system for synthesis of oligophenylalanine the antibiotic inhibits every round of peptide elongation, including dipeptide formation, to roughly the same extent.


The joining of the 30-S initiation complex with the 50-S subunit, the main target for thiostrepton. The study undertaken in this paper on the mode of action of thiostrepton provides data which permit a more precise localization of the main target of thiostrepton. There is severe impairment of the joining of the 50-S subunit, probably carrying thiostrepton, with either the 30-S subunit or the 30-S initiation complex. The degree of impairment of this coupling is temperature dependent, being almost completely inhibited at 0 degrees C, whereas at 37 degrees C the effect is much less marked, provided that natural messenger RNA is present. The inhibition of initiation by thiostrepton is more severe in the presence of IF-1, a factor, which similar to thiostrepton, is able to shift the dynamic equilibrium of 70-S in equilibrium 50-S + 30-S more towards dissociation. By means of 14C-labeled IF-2 it is demonstrated that the binding of IF-2 into the 70-S initiation complex is prevented by thiostrepton, which seems to be the main cause for non-coupling.


Preferential digestion of (A plus T)-rich stretches of yeast mitochondrial DNA in isolated mitochondria. Yeast mitochondrial DNA labelled in vitro by incubation of isolated mitochondria with DNA precursors exhibits skewed profiles on isopycnic CsCl gradients. The skew is not due to nuclear DNA nor to single-stranded mitochondrial DNA in the product labelled in vitro. Simultaneous labelling with [3H]dTTP and [14C]dGTP in vitro indicates a gradient of base composition in the DNA labelled in vitro. Thus, selective degradation of mitochondrial DNA occurs during incubation, converting large molecules having the mean density of mitochondrial DNA into smaller molecules of higher mean density and with higher G:T ratio. Similarly skewed distributions can also be produced by incubation of mitochondrial DNA labelled in vivo with the yeast mitochondrial fraction or with micrococcal endonuclease, an enzyme known to selectively hydrolyse (A plus T)-rich regions of DNA.


Interaction of RNA polymerase from Escherichia coli with DNA. Analysis of T7 DNA early-promoter sites. A method was devised for directing RNA polymerase on a single promoter site on T7 DNA. Initiation complexes were formed on each of the three main promoter sites using one dinucleotide plus one nucleoside triphosphate. The ternary initiation complexes are resistant to rifampicin action, to inhibition by (rI)n at 0 degrees C and are stable at high salt concentrations. A minimum of a trinucleotide is required to form a stable ternary complex. To determine which promoter site was selected by RNA polymerase during initiation, the (rI)n-resistant RNA was digested by RNAse III to generate three characteristic initiator RNA fragments, resolved by gel electrophoresis. The three major promoter sites could be selected individually by using different primer and substrate combinations ApC plus ATP selected promoter A3, CpG plus CTP selected A2 and CpC plus ATP specified preferentially A1. A number of primer-substrate combinations specified each site at low salt concentration but the substrate requirement became very stringent at high salt concentration, suggesting that the postulated local opening of the promoter site could be more or less extensive, depending on the ionic strength. The minimum opening observed at high salt concentration corresponded to the insertion of a leader trinucleotide sequence. The promoter region melted by RNA polymerase at low salt concentration was (G plus C)-rich and corresponded to about 9 to 11 base pairs. Sequences of the melting recognition regions were tentatively inferred from the results.


Immunologic chimerism as evidence of bone marrow graft acceptance in an identical twin with acute lymphocytic leukemia. Bone marrow from a well child was infused into her identical twin who had acute lymphocytic leukemia. In an attempt to provide an immunologic tag for use in twin transplantation, the donor twin was immunized to Keyhole limpet hemocyanin (KLH) and yellow fever virus prior to the marrow infusion. Subsequent immunological chimerism in the recipient twin gave evidence for graft acceptance.


Diffusion chamber culturing of haematopoietic cells: methodological investigations and improvement of the technique. Mouse bone marrow cells were cultured to determine some basic characteristics of the diffusion chamber (DC) technique. The main findings were as follows: (i) Incorporated 3H-thymidine was conveniently measured after deposition of a cell sample on glass fibre discs followed by methanol washing. Varying the specific activity from approximately 2-approximately 20 Ci/mmole did not affect the relationship between DC cellularity and isotope incorporation. Isotope uptake was similar regardless of whether 3H-thymidine of high or low specific activity had been used. (ii) Higher cell yields were obtained when Millipore or Acropor filters were heat-sealed rather than glued to the plastic rings, when Millipore filters were moistened before DC filling, when we omitted gluing the closing plugs, and when we used an average pore size of 0.22 mum for the DC walls rather than one of 0.10 mum or 0.45 mum. Varying the ring thickness from 2.0-2.5 mm did not impair the cell growth, nor did removal of a softening agent from the plastic rings improve it. (iii) More cells were retrieved from DC carried by young rather than by older mice. Results were not influenced by sex or strain differences between the donor and the host or by the number of implants, whether single or double, within an individual host. (iv) Adding Ficoll to the pronase solution increased the yield of viable CFU-C. (V) Diffusion rate through the DC walls declined with increasing period of culturing, so that i.p. 3H-thymidine is not a flash label for 7-day cultures, for example. The great variability of 3H-thymidine diffusion into i.p. DC was markedly reduced by in vitro exposure to the unopened DC to the isotope. (vi) DC could be incubated in vitro in a medium devoid of protein for at least 6 hours without a fall in 3H-thymidine incorporation rate or CFU-C content, provided that pH was kept constant.


Toxicologic and marrow transplantation studies in rhesus monkeys given dimethyl myleran. The hematologic and pathologic effects of single doses of dimethyl myleran (DMM) were evaluated in rhesus monkeys with and without the infusion of marrow. DMM 3 to 5 mg/kg produced severe but reversible myelotoxicity and no non-marrow toxicity. All animals given DMM 10 mg/kg died of marrow failure which was consistently reversed by the infusion of cryopreserved autologous marrow. At higher doses of DMM the protective effect of autologous marrow was less consistent due to increased gastrointestinal toxicity. Allogeneic engraftment was not achieved following administration of 10 mg/kg of DMM. In two animals receiving 15 mg/kg, proliferating allogeneic donor cells were present in recipient marrow seven days after marrow infusion.


Weakening of genetic resistance. I. The effect of injection of endotoxin, Freund's complete adjuvant and alloantiserum. A parent to F1 transplantation combination was used to study the weakening effect of endotoxin, Freund's complete adjuvant and alloantiserum on genetic resistance. The relationship between time of treatment with endotoxin, Salmonella typhosa, and Freund's complete adjuvant, and their weakening effect was assessed by use of the spleen colony technique. CFU growth studies revealed that both endotoxin and alloantiserum were capable of weakening genetic resistance in the spleen but were unable to induce weakening of the resistance in the femoral marrow cavity. These results led us to the conclusion that the agents might not have a direct effect on the effector cells of the resistance. The weakening induced by endotoxin and alloantiserum seemed to be related to a certain immunological phenomenon in the spleen. In this phenomenon macrophages are likely to play a role since a number of agents capable of weakening resistance were known for their capacity to influence the mononuclear phagocytic system.


The methylation of mercuric chloride by human intestinal bacteria. Most strains of staphylococci, streptococci, yeasts and E. coli isolated from human faeces, could synthesize methylmercury compounds. In contrast, few strains of obligate anaerobes could do so. Up to 6 ng methylmercury/ml were formed in 44 h from 2 mug mercuric chloride.


Erythrocyte agglutinins in the blood of certain ascidians. Plasma from Ciona intestinalis, Phallusia mamillata and Ascidia malaca possess hemagglutinin for a variety of erythrocytes. Results obtained by physical and chemical treatments suggest that hemagglutinin for Phallusia mamillata and Ascidia malaca may be a protein or a protein-like substance.


Oxygen tension: dependent or independent variable in local control of blood flow? We have looked at a fairly simple model of blood flow regulation at the microvascular level consisting of suffused microvascular preparations and isolated smooth muscles. When the O2 demand of the microvessel preparations was decreased by elevating suffusion solution PO2, changes in wall PO2 indicated that only the smallest microvessels could be controlled by a direct effect of oxygen. In vivo and in vitro studies of the oxygen sensitivity of smooth muscle indicated that even the smaller vessels were probably not directly controlled by oxygen availability during a free flow state, since measured perivascular PO2's did not fall to levels low enough to alter contractile performance of the vascular smooth muscle. Our data indicate that in any condition in which flow is interrupted for periods in excess of about 30 sec, one might anticipate vascular relaxation due to oxygen lack. It was judged resonable to extrapolate our findings to autoregulation of blood flow in resting skeletal muscle and to responses to modest exercise. Our data indicate that it is improbable that oxygen acts by a direct effect on the smooth muscle under these conditions. On the other hand, they suggest that a direct effect of oxygen might well be important in causing postocclusion hyperemia.


Biochemical function of selenium and its relation to vitamin E. Glutathione peroxidase (glutathione:H2O2 oxidoreductase, E.C. 1.11.1.9), isolated from ovine and bovine erythrocytes, has recently been shown to contain 4 selenium atoms per mole, an average of 1 Se per protein subunit of about 22,000 molecular weight. Selenium deficiency in the rat, chick and sheep causes dramatic decreases in the activity of this enzyme in the tissues, but certain sites such as liver are affected more than others. Decreases in glutathione peroxidase correlate with lesions caused by selenium deficiency and appear useful in diagnosing selenium deficiency. Glutathione peroxidase is an important enzyme in destroying H2O2 and organic hydroperoxides such as lipid hydroperoxides. It therefore guards against oxidative damage to the cell membranes and other oxidant-sensitive sites in the cell. While this selenium-dependent system destroys lipid hydroperoxides and other peroxides, vitamin E is believed to protect against oxidant damage to membranes by preventing the formation of lipid hydroperoxides. A scheme is proposed, based on oxidant damage and its prevention, which accounts for the interaction between selenium, vitamin E, unsaturated lipids, sulfur-containing amino acids, and cell damaging agents such as oxidant stressors and toxicants such as silver and tri-o-cresyl phosphate. The background for such a scheme is reviewed.


Mechanisms of action of selenium and vitamin E in protection of biological membranes. Opinions vary as to the mode of action of vitamin E and selenium. Some argue that they act as nonspecific biological antioxidants. Others propose that the functions of the two substances are distinct and that vitamin E acts as a true vitamin in addition to functioning as a lipid-soluble antioxidant. Support for the "Biological Antioxidant Theory" is largely circumstantial. However, lipoperoxides have been detected in adipose tissues of vitamin E-deficient animals, and increased rates of in vitro peroxidation have been demonstrated in homogenates of several tissues of selenium and vitamin E-deficient animals. The basis of the antioxygenic role of selenium in these systems was elucidated by the discovery of Rotruck et al. (1973) that selenium is a component of rat erythrocyte glutathione peroxidase. Further studies in this laboratory have demonstrated the important role of glutathione peroxidase in protection against the vitamin E- and selenium-deficiency disease of chicks, exudative diathesis, which results from increased capillary permeability. Also shown were the activities of both dietary selenium and vitamin E in prevention of ascorbate-induced peroxidation in mitochondrial and microsomal preparations from chick liver. Recent results demonstrate that both selenium and vitamin E are required to protect hepatic mitochondria and microsomes from peroxidative degradation. Dietary requirements of the chick for both nutrients for this function have been determined: approximately 0.06 ppm selenium in the presence of adequate vitamin E; 30-50 IU vitamin E per kg in the presence of adequate selenium.


In vitro production of the factors effective in the transfer of experimental autoimmune aspermatogenesis. We studied the biological effects of the factors released into the culture medium by the lymphocytes from animals sensitized with testicular antigen. The supernatants from cultures of these lymphocytes were active in vitro in the migration inhibition tests because they transferred the sensitivity to normal spleen cells. In vivo, they transferred aspermatogenesis in the allogeneic and xenogeneic system. The results suggest a specific effect on the behaviour of macrophages rather than a direct cytotoxic effect of the active factors.


Inhibition of haemopoietic stem cells by syngeneic lymphocytes treated with antilymphocyte serum. Some aspects of the mechanisms by which antilymphocyte serum, its globulin fraction, or phytohaemagglutinin act to reduce the stem cell content of murine haemopoietic tissues were studied, using the method of clonal proliferation of stem cells in the spleens of lethally irradiated (830R) mice. The transplants were treated in vitro with ALS, ALG or PHA for 30 min. at 37 degrees C. Direct treatment of spleen or bone marrow cells with ALS or PHA resulted in pronounced inhibition of the colony-forming capacity of haemopoietic tissues in syngeneic irradiated recipients. The degree of inhibition was dependent on the concentration of ALS or PHA. It was found that the inhibition of CFC with ALS was mediated by lymphoid cells: bone marrow CFC were reduced after 30-min incubation at 37 degrees C with syngeneic lymph node or thymus cell pretreated with ALG and washed free of its excess. It was further demonstrated that ALG-treated thymocytes released, upon 30-min. incubation at 37 degrees C, some material capable of inhibiting bone marrow CFC. No release was observed with PHA-treated thymocytes. Some properties of the active supernatants are described and it is suggested to designate the observed inhibitory activity as Stem-cell Inhibitory Factor.


Analysis of the Kinetics of the immune response by means of ionizing radiation. The inhibitory kinetics of the primary immune response was studied by means of lethal irradiation. CBA mice were immunized with sheep red blood cells and 48, 72 and 96 hours later, they were irradiated with 750R. Afterwards, the number of direct plaque-forming cells was determined in their spleens during 58 to 66 hours at 4-hour intervals. After irradiation at 48 and 72 hours of the immune response, a plateau of different duration was obtained: 48 hours in the former and 24 hours in the latter case. In both cases, reduction in the PFC number began at about 96 hours of the resonse. The PFC reduction curves in these two series of experiments and those obtained 96 hours after irradiation and in control, non-irradiated mice were parallel. The mean half-time of the PFC number reduction as determined by these curves was 6 hours. The time of functioning of PFC in the spleen is discussed with respect to recruitment of proliferating precursors.


Effects of various immunosuppressive drugs on the development of experimental renal infection in rats. The effect of azathioprine, hydrocortisone and antihymocyte serum on the development of experimental colibacillary renal infection in rats was assessed by macroscopical findings in the kidneys, especially by determining the number of microbes in the infected organ, on day 7 after injection of the infectious agent. The greatest multiplication of bacteria and so the most pronounced macroscopical changes occurred after ALS treatment. There was statistically significant difference between ALS-treated group and the control group, whereas the differences between controls and the other experimental groups (Imuran, hydrocortisone) were non-significant.


Effect of intracisternal insulin on plasma glucose and insulin in the dog. The intracisternal administration of insulin (0.2 U./kg.) to anesthetized dogs resulted in an increase of arterial immunoreactive insulin and a decrease of plasma glucose relative to a control injection. The arterial responses were significantly attenuated when the insulin was administered to the cisternum of subdiaphragmatically vagotomized dogs. When cerebrospinal fluid glucose was lowered by injecting pneumococcal neuraminidase intracisternally, no peripheral hyperinsulinemia resulted, indicating that increased spinal fluid insulin and its consequent increase of glucose uptake, rather than decreased spinal fluid glucose, is necessary to elicit the vagally mediated insulin secretion and hypoglycemia. It is hypothesized that increased spinal fluid insulin causes an increased glucose uptake of some glucoregulatory area of the brain and that the elicited reflex is vagally mediated pancreatic insulin secretion.


Dynamics of insulin release and microtubular-microfilamentous system. VII. Do microfilaments provide the motive force for the translocation and extrusion of beta granules? The active role played by beta-cell microfilamentous structures in the dynamics of insulin secretion was investigated by examining the influence of cytochalasin B upon various parameters of hormonal release by the isolated perfused rat pancreas. The view that the cytochalasin-induced changes in insulin release are due to a primary biophysical effect on microfilaments, rather than to an unrelated biochemical alteration of the beta-cell glucose-sensor device, was strengthened by the following observations: (1) the onset and disappearance of the cytochalasin B-induced facilitating action upon insulin release followed a time-course parallel to that characterizing the ultrastructural changes provoked by the drug in the distribution of beta-cell microfilamentous material; and (2) cytochalasin B facilitated leucine-induced insulin release in the presence of a very low glucose concentration. The mold metabolite was also found to transform transient secretory responses into biphasic ones and to prevent the reduction that normally affects the early response to insulinotropic agents when the pancreas is stimulated a few minutes after a prior and short exposure to glucose. The release of insulin evoked by either glucose or gliclazide was abolished in the absence of extracellular calcium, whether in the presence or absence of cytochalasin B. Theophylline and cytochalasin B exerted a synergistic effect upon glucose-induced insulin release. These data support the concept that calcium-dependent contractile events involving cytochalasin B-sensitive microfilamentous structures provide the motive force for both the intracellular translocation and exocytotic release of beta granules.


Effect of cerebral intraventricular insulin on pancreatic insulin secretion in the dog. The effect of cerebral intraventricular insulin on pancreatic insulin secretion was investigated. An extracorporeal pancreatic blood circuit was established after laparotomy to monitor blood flow and insulin concentration directly from the superior pancreaticoduodenal vein. Phentolamine was infused throughout (0.2 mg./min. intravenously) to block alpha-adrenergic effects of any catecholamine secretion induced by surgical stress. Glucose (1.5 mg./kg./min. intravenously) was infused to maintain a constant baseline stimulation of insulin secretion. Six dogs received insulin and six control dogs received saline through a spinal needle stereotaxically placed into the left lateral cerebral ventricle. After central injection of insulin (0.2 U./kg.) there was a significant increase of pancreatic output as early as five minutes. It is concluded that the pancreatic beta-cells are under the influence of insulin-sensitive cells of the CNS.


Ethanol-induced alterations of glucose tolerance, postglucose hypoglycemia, and insulin secretion in normal, obese, and diabetic subjects. Ethanol at an average blood concentration of 1 mg. per milliliter enhanced the immediate (first-phase) and prolonged (second-phase) insulin response to an intravenous glucose load in nonfasting normal human subjects. Simultaneously, the glucose disposal rate was increased and the postglucose hypoglycemia was accentuated, resulting in definite hypoglycemic symptoms in some individuals. Oral glucose tolerance was not changed by ethanol administration, but the thirty-minute blood glucose and plasma insulin values were increased, suggesting that alcohol might accelerate the absorption of glucose from the gut. Ethanol given orally during evening hours (1.5 gm. per kilogram) caused a nocturnal hyperinsulinemia and a decrease of blood glucose, but not an actual hypoglycemia. Oral glucose tolerance and plasma insulin response tested the next morning, when ethanol had disappeared from the blood, were not influenced by drinking the previous evening. The K-value of intravenous glucose was increased at this time, however. When alcohol was administered for one week at a dose corresponding to 25 per cent of daily calories and substituting for fat, both the oral and intravenous glucose tolerances were impaired in each subject but the insulin response remained unchaged. In obese nondiabetic subjects, ethanol did not potentiate the early insulin response to intravenous glucose but it increased the second phase of insulin secretion in response to sustained hyperglycemia. In contrast to conditions in nonobese subjects, the glucose disposal rate was not incresed and postglucose hypoglycemia was not accentuated by ethanol in overweight subjects. In insulin-deficient diabetic patients the absent early insulin response could not be restored by ethanol, and the late component of insulin release was little increased by alcohol infusion. Ethanol did not improve the glucose utilization of diabetic patients.


The G cell population of the pyloric antrum of the cat. The method used in this study to quantify the G cell population of the cat is similar to the technique used for measuring the parietal cell mass in the stomach of rats and man. The method may be used on the human antrum. Mucosal sampling technique and immunofluorescence methods are described. The method is reproducible. G cell distribution in the antrum was not uniform and the highest concentrations were found along the lesser curvature. The G cell population was 10.27, 12.66, 12.87, 12.88 and 15.42 million cells (mean 12.8 x  10(6)) per antrum in each of the five cats studied.


Hepatocellular uptake of bile acids in the dog: Evidence for a common carrier-mediated transport system. An indicator dilution study. The purpose of this study was to compare the uptake of taurocholate (TC) by the liver of the dog to that of taurochenodeoxycholate (TCDC) and to examine the influence of TCDC on TC uptake. The uptake of these bile acids by the liver of the intact was studied with the multiple indicator dilution method, using 51Cr-labeled red blood cells as a vascular indicator, 125I-labeled albumin as an extravascular reference, and one or two labeled bile acids. The analysis of the dilution curves fitted well a three-compartment model, and no return of the extracted bile acids to the extracellular space could be detected. The initial space of distributin of TC in the presence of a fixed dose of TCDC was 1.25 +/- 0.09 (SD) times greater than that of albumin. Analysis of the data for TC uptake in the presence of TCDC was consistent with Michaelis-Menten kinetics, as previously established for TC alone. The calculated initial maximal velocity of uptake (Vmax) of TC in the precence of TCDC was 4.92 +/- 1.36 (SD) mumoles per sec per 100 g of liver. This value is not significantly different from the Vmax previously calculated for TC alone. The apparent KD (dose of TC yielding half-maximal velocity) was 18.10 +/- 6.14 (SD) mumoles per 100 g of liver, a value significantly higher than that obtained in the absence of TCDC. A K1 (the Michaelis-type constant characterizing the inhibition of TC uptake by TCDC) of 2.71 +/- 1.08 (SD) mumoles of TCDC per 100 g of liver was calculated. These observations are consistent with competitive inhibition of TC uptake by TCDC, and thus with the existence of a common carrier-mediated transport system for TC and TCDC. Estimation of the kinetic characteristics of the two bile acids from dilution curves obtained simultaneously showed that the affinity for the carrier was higher for TCDC than for TC, but that the maximal capacity was lower for TCDC than for TC.


Serum immunoglobulin M concentrations following bowel resection in chronic inflammatory bowel disease. Serum immunoglobulin alterations were characterized in 20 patients with chronic inflammatory bowel disease (CIBD) undergoing bowel resection and in 11 control subjects undergoing intra-abdominal operations. Serum immunoglobulin M (IgM) values became elevated during the 2nd week in both groups with individual serum IgM concentrations reaching as high as 9 times preoperative values. The IgM increment, which was 19 S polyclonal immunoglobulin, was significantly greater in the CIBD group than in controls. Because of a recent suggestion that this IgM rise might be related to the pathogenesis of CIBD, the affinity of this IgM for colon was studied in two ways. Serum anticolon antibody titers did not change after surgery nor did IgM, isolated from serum after surgery and labeled with 125I, bind to homogenates of normal or CIBD colon. Titers of heterophile antibody and antibodies directed against Escherichia coli, Bacteroides, and blood group antigens increased in many patients. In most instances, these increments were shown to be antibody of the IgM class by reduction with 2-mercaptoethanol. These data indicate that the IgM changes after abdominal surgery (1) are not specific for CIBD patients, (2) do not represent antibody directed against colon antigens, and (3) include increased titers of several IgM antibodies. These findings might be explained by a response to bacterial or tissue antigens released at or after the time of surgery.


[On the beginnings of hormonal contraception (author's transl)]. The possibility of hormonal contraception was postulated as early as 1919 by the physiologist Ludwig Haberlandt in Innsbruck. The same year, he began to test his hypothesis in animal experiments. In 1924 he succeeded in his efforts to render mice infertil by orally administering ovarian and placental extracts. He failed to have his method tried in women.


The autonomic nervous system and its relationship to tubal ovum transport--a reappraisal. The role of the autonomic nervous system in controlling ovum transport remains obscure. Although not studied extensively in the oviduct, the para-sympathetic nervous system does not appear to significantly influence ovum transport. The sympathetic nervous system of the oviduct and its pharmacology have been studied more thoroughly. Despite this, little information is available concerning cellular mechanisims of adrenergically altered motility or transport. In spite of much speculation, the weight of evidence suggests that, at least in the rabbit, the sympathetic nervous system plays a minor role in the control of ovum transport.


Sperm transport to and survival in the human fallopian tube. A review is given on sperm migration to and sperm survival within the human Fallopian tube. Sperm migration from the external os can be very fast. The survival time of spermatozoa in the oviduct has been demonstrated to be 85 h. Spermatozoa normally enter the abdominal cavity through the open fimbriated end. Laterally closed oviducts retain spermatozoa resulting in a larger number of spermatozoa than in the normal oviduct, where the number of sperm at the site of fertilization is very low. The morphology of spermatozoa reaching the ampulla of the oviduct is mostly normal, which seems to be based on the correlation between normal morphology and good motility. Spermatozoa within the abdominal cavity do not cause antibody formation of any importance for the fertility of the woman.


The isolation of protoplasts of the fission yeast Schizosaccharomyces by Trichoderma viride and snail enzymes. The formation of protoplasts of the fission yeasts Schizosaccharomyces pombe and Schizosaccharomyces versatilis after the combined application of snail enzymes and Trichoderma viride enzymes in an osmotic stabilizer (0.4M KCl, pH 5.5) was studied by light and electron microscopy. The effect of the enzymes used leads during 30 min to the formation of 100% protoplast population. Using electron microscopy no original walls or wall remnants were detected in the suspension of protoplasts. Protoplasts are viable and in liquid nutrient medium they regenerate cell walls and revert into normal cells. Such a protoplast population may be useful for biochemical study of protoplast metabolism by quantitative methods as well as for the chemical study of regenerating cell walls.


Penicillinamidohydrolase in Escherichia coli. II. Synthesis of the enzyme, kinetics and specificity of its induction and the effect of O2. The differential rate of synthesis of penicillinamidohydrolase (penicillin acylase -- EC 3.5.1.11) was studied in Escherichia coli growing in some chemically defined media and in a complex medium. The enzyme is synthesized at a constant rate only during the exponential phase of growth of cells. Its synthesis is induced most effectively (with respect to quantity) by phenylacetic acid. The induction lag of the enzyme synthesis in a medium with acetate corresponds to two generation times. The highest rate of the enzyme synthesis is reached in a medium containing phenylacetic acid as the only source of carbon and energy. The enzyme synthesis is fully repressed by an increased concentration of dissolved oxygen in the medium, even when Escherichia coli is cultivated in the medium with phenylacetic acid as the only carbon and energy source.


The effect of medium composition on the production of valine by Corynebacterium 9366 - EMS/184. The auxotrophic mutant Corynebacterium 9366-EMS/184, requiring isoleucine accumulates about 22 g of L-valine per litre after a 96 h cultivation in a medium containing 15% sucrose and 1% ammonium acetate.


Formation of a complex between yeast proteinases A and B. Sephadex G-100 gel filtration of a purified mixture of tryptophan-synthase-inactivating enzymes I and II from Saccharomyces cerevisiae, which had previously been identified as yeast proteinases A and B, yields two coincident peaks of tryptophan-synthase-inactivating activity and proteinase A and B activities. The peaks correspond to molecular weights of approximately 100 000 and 50 000. It could be demonstrated, with purified proteinases A and B, that the high molecular weight peaks is due to the formation of a stoichiometric complex between proteinases A and B. The AB complex can easily be separated into its constituent enzymes by DEAE-Sephadex chromatography. When in the AB complex, proteinase B molecules still bind the specific proteinase B inhibitor from yeast.


Synthesis of a chemically reactive analog of the nonsense codon U-G-A. Its reaction with ribosomes of Escherichia coli. Nitrophenylated 5'-uridylic acid could be employed as primer in a polyribonucleotide nucleotidyltransferase (Micrococcus luteus) reaction to yield 5'-nitrophenylated Pu-G-A. After reduction of the nitrophenyl moiety and subsequent bromoacetylation, a 5'-bromoacetamido-phenyl-phosphorylated U-G-A was obtained, which could be used as an affinity label for the ribosomal binding site of the nonsense codon. If freshly prepared active ribosomes were employed in the incubation mixtures, the U-G-A analog reacted exclusively with one protein, which is tentatively identified as protein S18. Exposure of ribosomes to low temperatures gave rise to a reaction of the U-G-A label with another protein, which was identified as protein S4, the ram gene product. The results of the affinity labeling experiments with the chemically reactive U-G-A derivative are very similar to that obtained with a corresponding derivative of the initiation codon A-U-G (O. Pongs and E. Lanka (1975) Proc. Natl. Acad. Sci. U.S.A. 75, 1505-1509), which suggests that 70S ribosomes have one preferential codon binding site.


The primary structure of the 5s rRNA binding protein L25 of Escherichia coli ribosomes. The primary structure of protein L25 from the large subunit of Escherichia coli ribosomes was determined by isolation and analysis of peptides obtained after cleavage of the protein with trypsin, thermolysin and Staphylococcus protease as well as by Edman degradation of the intact protein and of a CNBr peptide. The complete amino acid sequence is shown in Fig. 4. There are sequence homologies within protein L25 (Table 6) as well as between protein L25 and other ribosomal proteins (Table 5).


Properties of Isoaccepting tRNALys, tRNAGlu and tRNAGln from rabbit Reticulocytes and liver. Multiplicity, Codon Recognition and Inactivation by Iodine. The three tRNA species from rabbit liver and reticulocytes, each corresponding to the codons XAA and XAG(X equals A, G, C) were investigated. The elution patterns of the isoaccepting tRNA subspecies after separation by reversedphase chromatography followed by determination of amino acid acceptance have common and differing characteristics. Three subspecies of tRNALys, two of tRNAGlu and three of tRNAGln have been found in reticulocytes, wheras two subspecies of tRNALys, three of tRNAGlu and four of tRNAGln have been determined in rabbit liver. Examination of codon recognition by means of a ribosome binding assay showed that each subspecies binds exclusively in the presence of only one of the two possible triplets; i.e. wobbling was not found. It has been assumed that this high specificity, which is not to be expected according to the "wobble hypothesis", is due to a 2-thiouracil base in the first position of the anticodon. This was supported by oxidation experiments with iodine. Treatment with iodine significantly reduces the aminoacylation capacity of all subspecies that show specific binding with triplets ending in adenosine. We describe and compare for the first time the characteristics of eucaryotic tRNA species which occupy homologous positions in the codon table. Thus it can be seen that all the investigated subspecies have rigorously specific codon recognition in common, wherby "wobbling" can probably be excluded by a principle that is valid for all the investigated species.


B-chain shortening of matrix-bound insulin with pepsin, II. Preparation and properties of camel des-pentapeptide (B26-30)- and des-PheB1-des-pentapeptide (B26-30)- insulin. Digestion of matrix-bound camel insulin with pepsin was found to be restricted to the cleavage of the peptide bond between phenylalanine(B25) and tyrosine(B26). Purification of the camel des-pentapeptide(B26-30)-insulin obtained was achieved by gel filtration and ion exchange chromatography, yielding a molecularly uniform product. In the same way, camel des-PheB1-insulin prepared according to a method described for bovine insulin was hydrolyzed with pepsin and purified to give the des-PheB1-despentapeptide(B26-30)-insulin. The biological activity of these modified camel insulins was found to be as much reduced as the corresponding bovine insulin analogues. However, the reduced biological activity of camel des-PheB1-insulin (60%) was a contrast to the fully active bovine des-PheB1-insulin.


Pathology of cerebral embolization caused by nonthrombotic agents. Although apparently infrequent, a variety of agents other than thrombotic material are known to cause cerebral embolization, often with serious consequences. The emboli may originate in different parts of the body or may be introduced from outside under diverse circumstances. The pathologic aspects of these individual embolic phenomena are described with particular emphasis on their recognition at autopsy. Adequate autopsy study appears to be the most important source of information for further elucidation of the incidence, mechanism, and sequelae of these heterogeneous embolic lesions of the brain. Such information is essential for clinical evaluation, management, and, more importantly, prevention of some of these potentially serious embolic phenomena.


Vascular structures in brain tumors.

Vascular structures in brain tumors. The blood vessels within brain tumors show alterations from the normal anatomy. Some of these seem to be related to an increased capacity to transfer materials between the lumen and the parenchyma and are probably intimately connected with the edema associated with the tumor. These alterations include fenestration, widened intercellular junctions, increase in pinocytotic vesicles, and infolding of the luminal surface. Other alterations are observed but their function is not as clear. The latter include an increase in the number of tubular bodies, the appearance of tubular structures within vacuoles, tubular arrays within the nuclear envelope and rough endoplasmic reticulum, and endothelial proliferation, among others.


Distribution of cytophilic and anti-macrophage antibody on the macrophage surface. The binding of cytophilic antibody to the macrophage surface results in patterns of distribution similar to those obtained when specific anti-macrophage antibody is used. These patterns are (a) diffuse circumferential binding at 4 degrees C, and (b) capping at 37 degrees C. The non-random distribution of the cytophilic antibody suggests that it binds to certain well-defined sites on the macrophage membrane.


Endocytosis of chlorambucil-bound anti-tumor globulin following "capping" in EL4 lymphoma cells. A rabbit 7 S antibody reacting specifically with a tumor assoicated antigen on the surface of mouse EL4 lymphoma cells could form "caps" and subsequently undergo endocytosis even when the antibody was non-covalently bound to chlorambucil. As the alkylation of nuclear DNA appears to be the basis of tumor inhibition by alkylating agents like chlorambucil, facilitation of the transport of chlorambucil across cell membrane by anti-tumor antibodies might explain, at least in part, the increased tumor inhibition by cholorambucil bound anti-tumor antibodies compared to tumor inhibition by equivalent amounts of tumor antibodies or chlorambucil alone.


Studies on B- and T-cell receptors for lysozyme. BALB/c mice have been immunized by intravenous administration of native or reduced and alkylated lysozyme. Primary immune response to these antigens was studied at the humoral level (by the Farr assay) and at the cellular level (by the rosette and the plaque assays using lysozyme coupled to sheep or pigeon erythrocytes). Antibodies and theta-negative RFC were specific for the antigen used for immunization. Specific inhibition of theta-negative RFC after incubation with the soluble immunizing antigen confirmed this specificity. Conversely, most theta-positive RFC had double specificity both to native and denatured lysozyme and showed lower avidity for the immunizing antigen, as shown by inhibition studies with soluble antigen. These data suggest that T cells have a broader specificity for lysozyme than B cells and also that, in this particular system, cytophilic antibodies are probably not responsible for the formation of theta-positive rosettes.


Age-dependent changes in the relative affinity of anti-dinitrophenyl antibodies in mice. The affinity of anti-DNP antibodies produced by mice of various ages has been studied at the cellular level by the plaque inhibition technique. The affinity of PFC produced shortly after a single injection of antigen was found to increase during the first 6 weeks of life. The responses of 1- and 2-week-old animals also showed apparently restricted heterogeneity. The difference in the affinity of anti-DNP--PFC between young and adult mice could not be attributed to different (serum) levels of antigen or to differences in the rate of maturation of affinity during the immune response. Cell transfer experiments suggested that the age-dependent increase was due to a change in the population of antibody-forming (B) cell precursors and not to a progressive improvement of T-cell function. This findings is interpreted as favouring somatic mutation theories of antibody diversity.


Experimental murine leprosy: growth of Mycobacterium lepraemurium in C3H and C57/BL mice after footpad inoculation. Forty-three female C57/BL and C3H mice were inoculated with 2.7 X 10(6) Mycobacterium lepraemurium into each hind footpad. The foot thickness and the number of acid-fast bacilli in the footpad and popliteal and inquinal lymph nodes were recorded. In addition the morphological index and the mean bacillary length were determined in the footpad and in the popliteal lymph node. The bacilli multiplied in both strains during the first 4 weeks after inoculation. After that time no further increase in acid-fast bacilli was observed in the C57/BL strain; the bacilli became elongated and the morphological index decreased. These changes were preceded by a local swelling of the footpad due to the onset of an immune reaction. Thus, under the present conditions, C57/BL mice were able to resist experimental infection with M. lepraemurium by developing an immune response. In C3H mice no indication of an immune reaction was detected, and the bacilli continued to multiply throughout the observation period. The mouse footpad model seems to provide an excellent basis for the use of experimental murine leprosy to study immunity to mycobacterial infections. Certain aspects of the present model are discussed in relation to the mouse footpad model as used in the study of M. leprae infection in mice.


Induction of the immune response to cell surface antigens in vitro. Two tissue culture incubation systems are described in which immune responses to cell surface antigens have been demonstrated. In the one-way "mixed lymphocyte interaction" system, a specific stimulation of thymidine uptake was induced by a particulate membrane antigen fraction, the microsomal lipoproteins (MLP), when low levels (0.01 to 0.001 mug per ml) were incubated with spleen or lymph node cells from nonsensitized mice. No stimulation was seen when allogeneic MLP was used at high levels, 10 mug per ml, nor at any level with syngeneic MLP. Specific effectors were demonstrated after 72-hr incubation with stimulatory levels of allogeneic MLP in three separate in vitro assays, a plaque-forming cell reduction assay, a tumor target assay, and an antigen-binding cell assay. In the latter assay, [125I]MLP was used as the source of antigen. This system has limited potential inasmuch as mouse spleen cells do not survive in it beyond the 4th day of culture. The second tissue culture system, the Marbrook system, has much greater possibilities because at least 25% of the inoculum is recovered 7 days later. In this culture system a cell-free sheep erythrocyte membrane preparation can induce plaque-forming cells in the absence of macrophages. Using a sensitive radioimmunoassay, free specific antibody was detected in culture supernatant fluids. With the same culture system, allogeneic lymphocytotoxic cells (killer) have been induced with spleen cells from unprimed mice in strains differing at the major histocompatibility locus (H-2). Allogeneic MLP induced very significant "killer" cell activity with spleen cells from primed mice. In a syngeneic tumor system, significant amounts of killer cell activity were induced with unprimed spleen cell inocula, and much larger amounts induced with spleen cells from immunized mice.


Comparison of leukocyte count and function in smoking and nonsmoking young men. Leukocyte function and other hematological measurements were tested in 14 smoking and 13 nonsmoking young men free of intercurrent or chronic diseases. Leukocyte chemotaxis was depressed in smoking subjects when compared to the same subjects abstaining from cigarettes or to the nonsmokers. Smoking did not affect the whole blood bactericidal capacity of leukocytes and serum for Staphyloccus aureus or Klebsiella pneumoniae. Total leukocyte counts, hematocritis, and monocyte counts were higher in the smoking subjects when compared to the nonsmokers.


Klebsiella pneumoniae and Staphylococcus aureus infection in mice: difference in uremia and ammoniagenesis. Lethal infections by Staphylococcus aureus and Klebsiella pneumoniae were compared for kidney-related effects in mice. K. pneumoniae caused uremia and an increase in blood ammonia that could reach 2.5 times normal. These events did not occur in mice inoculated with S. aureus. Use of germfree animals indicated that most of the increase in ammonia arose from the gut, presumably due to greater availability of urea and ureolysis. Injected ornithine restored blood ammonia to nearly normal levels and extended survival.


Model system for studying colonization and growth of bacteria on a hydroxyapatite surface. A model system for the study of bacterial colonization and growth on a hydroxyapatite (HT) surface is described. Hydroxyapatite was crystallized over the surface of porous glass beads. Chemical analysis of the product showed that the ratio of Ca2+/P042- was indistinguishable from that of commercial HT powder. X-ray diffraction analysis supported the conclusion that the product was HT. A system employing [14C]polyethylene glycol, which selectively adsorbs to the glass surface of the beads, was developed to determine the amount of glass surface covered by HT. Over 90% of the glass surface could be covered by our method. The product, HT beads, consisted of approximately 20% (dry weight) HT. The HT beads possess several properties which make them potentially useful for studying microbial adherence, growth, and interactions. These include: (i) chemical similarity to the tooth surface, (ii) large surface area, and (iii) high density. We also describe a method for direct measurement of the microbial mass of cells growing on    beads. The method entails immobilizing a sample on a membrane filter (Millipore), staining it with amido black dye, and eluting the dye for spectrophotometric measurement. Streptococcus mutans served as the test organism. For free-growing bacteria the values measured with the filter assay were directly proportional to cell number, with a value of 1 mug of "protein" corresponding to about 1.5 X 10(6) colony-forming units, determined by viable count. For bacteria colonizing the beads, 1 mug of protein corresponded to about 2 X 10(7) colony-forming units on the beads during logarithmic growth. As the culture approached stationary phase, the efficiency of the assay decreased. These data indicate that multiple random samples, taken at a given time, are representative of the entire culture.


Hepatitis B antigens in serum and liver of chimpanzees acutely infected with hepatitis B virus. We report the temporal patterns of various immunohistological and serological parameters of acute self-limited hepatitis B virus infection of two chimpanzees, and we provide evidence that the synthesis of hepatitis B core antigen precedes that of hepatitis B surface antigen. Our data suggest that existence of a biphasic hepatitis B virus infection involving a hematogenous reinfection of the liver and indicate that recruitment of liver cells to produce hepatitis B virus may occur in a pattern consistent with a replicative cycle of about 8 days.


Plasmid-controlled colonization factor associated with virulence in Esherichia coli enterotoxigenic for humans. An enterotoxin-producing strain of Escherichia coli isolated from a case of cholera-like diarrhea (E. coli strain H-10407) was found to possess a surface-associated colonization factor. Colonization was manifested as the ability of small inocula (10(5) bacteria) to attain large (10(9)) populations in the infant rabbit intestine with a concomitant diarrheal response. A laboratory-passed derivative of E. coli H-10407, designated H-10407-P, failed to exhibit an increase in population in the infant rabbit and also failed to induce diarrhea. Cell-free culture supernatant fluids of E. coli H-10407 and H-10407-P produced equivalent enterotoxic responses in infant and in adult rabbits. Specific anti-colonization factor antiserum was produced by adsorbing hyperimmune anti-H-10407 serum with both heat-killed and living cells E. coli H-10407-P. This specific adsorbed serum protected infant rabbits from challenge with living E. coli H-10407 although the serum did not possess bactericidal activity. The anti-colonization factor serum did not agglutinate a strain of E. coli K-12 possessing the K88 colonization factor peculiar to E. coli enterotoxigenic for swine. By electron microscopy it was demonstrated that E. coli H-10407, but not H10407-, possessed pilus-like surface structures which agglutinated with the specific adsorbed (anti-colonization factor) antiserum. E. coli H-10407 possessed three species of plasmid deoxyribonucleic acid, measuring 60 X 10(6), 42 X 10(6), and 3.7 X 10(6) daltons, respectively. E. coli H-10407-P possessed only the 42 X 10(6)- and the 3.7 X 10(6)-dalton plasmid species. Spontaneous loss of the specific H-10407 surface-associated antigen was accompanied by loss of the 60 X 10(6)-dalton species of plasmid deoxyribonucleic acid and loss of colonizing ability. Thus, it is concluded that the E. coli colonization factor described here is a virulence factor which may play an important and possibly essential role in naturally occurring E. coli enterotoxic diarrhea in man.


Amount and avidity of antibody to Escherichia coli O antigen measured with the ammonium sulphate precipitation technique in children with urinary tract infections. The antibody amounts and avidities were analyzed in 13 patients with acute primary pyelonephritis, 11 patients with acute primary cystitis, and one with ureterocele and recurrent infections, using the ammonium sulphate precipitation (ASP) technique. The ASP titrations did not discriminate as well between pyelonephritis and cystitis as do the determinations with the indirect hemagglutination technique. The increase of antibody titer and avidity detected by the ASP method in some of the cystitis patients suggested a deeper tissue involvement in some cases resulting in antibodies demonstrable with this technique. Since control patients also showed a somewhat heterogenous antibody response regarding titer and avidity it cannot be excluded that stimulation by Escherichia coli antigens in the gut is detected by the ASP method.


Immunoglobulin-bearing lymphocytes: their demonstration in adult sheep and ontogeny in the sheep fetus. A method for the preparation of lymphocytes from sheep blood is described. Lymphocytes from adult and fetal sheep have been examined by immunofluorescence for surface immunoglobulin (sIg) determinants. Young adults (1.5 years) had a mean of 23.2% sIg cells, older adults (8.75 years) a mean of 9.7% sIg cells. Pregnancy did not influence these values. The earliest lymphocytes with sIg in fetal lambs were demonstrable at 52 days (96 mm crown-rump length) and countable by 56 days (110 mm CRL) at 0.3% sIg. The percentage rose rapidly to 15.1% between 78 and 87 days, followed by a fall to 2.1% around 117 days with a second increase thereafter. The significance of this spontaneous appearance of fetal sIg cells is discussed.


[A system to aid decision-making. Application to automatic interpretation of vectorcardiograms]. This paper presents a complete and autonomous system for the automated diagnosis (heuristic approach). The system was worked out by means of a small computer. A flexible, evolutive, quasi-universal system is achieved through original procedures. A specialised language enables the users to describe diagnoses and their criteria in symbolic form. A suitable compiler translates this symbolic writing into an interpretable object program. This, with the Interpreter program, constitutes the 'Automated Diagnosis Program'. Our data are vectorcardiograms recorded according to the Frank orthogonal system. After an interactive pre-processing process, 180 parameters--mostly spatial--are computed. The data, the computed parameters and additional information are stored in a data bank. Finally, the medical interpretation is automatically selected from 125 possibilities. The user could also utilise a data bank interrogation language.


Attitudes toward alcohol and drug use and abuse. I. Demographic and correlational data. Research concerning drug- and alcohol-related attitudes was reviewed. Demographic variables were not generally consistently related to attitudes. Most personality variables were also not consistently correlated with attitudes.


Chronic cerebral effects of alcohol and drug abuse. A minority of alcohol abusers develop severe cerebral dysfunction in the form of Wernicke-Korsakoff syndrome. There is also evidence to suggest that cerebral dysfunction, particularly impaired abstracting ability, occurs in that larger population of heavy drinkers who do not go on to develop the  Wernicke-Korsakoff syndrome. There is no consistent evidence that long-term marijuana, hallucinogen, or sedative use causes lasting neuropsychological disturbance. The deficits in abstract thinking reported by some LSD studies are similar to deficits others have reported among alcoholics. Since the LSD studies were not controlled for alcohol use, their interpretation is difficult. It appears that cerebrovascular accidents occur more frequently and at a younger age among amphetamine abusers. There is no reliable information about possible other long-term effects of stimulants on the brain per se (i.e., nonvascular complications). Abuse of intravenous narcotics has been associated with case reports of transverse myelitis and encephalitis. It is not known whether this pathology is a direct or hypersensitivity effect of narcotic drugs, of adulterants, or of infection.


The use of tranquilizers in the treatment of mixed drug abuse. If the abuse potential of tranquilizers, and particularly benzodiazepines, is examined by their administration as a therapeutic agent to persons dependent on alcohol, alcohol and drugs, or drugs alone, these substances are found to be potentially dependence producing with similar mechanisms to those of other hypnotics and sedatives. However, their abuse potential varies from one substance to another, and on the whole is relatively small. At all events, with constant medical supervision, patients who were previously dependent on a drug of barbiturate type do not independently increase their intake so as to become dependent on the tranquilizers. These experiments on persons with an inherent susceptibility to dependency show that tranquilizers must represent only a very small risk to the general population. The view that benzodiazepines are not potentially dependence producing has no more scientific foundation than the exaggerated fears of this potential. Important is that the uncontrolled supply of highly potent tranquilizers, and above all those with a strong sedative component, leads to dependence in a considerable percentage of cases in the treatment of mixed abuse. No conclusions in absolute terms on the risk presented by tranquilizers (and above all benzodiazepines) may be drawn from observations made in the uncontrolled administration of these substances in case of polytoxicomania. It follows that tranquilizers, which, as far as their therapeutic effect is concerned, are a valuable aid in the treatment of drug abuse, can be safely given when there is constant medical supervision combined with psychotherapy. The basic principle to be followed in the use of tranquilizers for the treatment of drug-dependent persons, is, therefore, their periodic administration in limited doses with constant medical supervision. In principle, however, because of the abuse potential which is nevertheless present, the same mode of administration is also to be recommended for other patient groups. Uncontrolled long-term administration is to be rejected in all circumstances, and supply should be strictly on prescription only in order to prevent uncontrolled acquisition. The various experiments carried out by us on the administration of benzodiazepines to patients with simple or mixed drug dependence show that the habit forming potential of benzodiazepines must, on the whole, be considered as being relatively small; at all events, this group of substances is the least hazardous of all known sedatives. This is of considerable importance, since the neuroleptics and antidepressants (which in themselves present no risk of abuse) are generally poorly tolerated and refused by patients with an inherent tendency towards drug dependence.


Effects of Demetrin on circulation.

Effects of Demetrin on circulation. In a double-blind study the influence of Demetrin on cardiovascular activity was investigated. Basic values before treatment were compared with values after 1 and 2 weeks' administration. The cardiovascular parameters "pulse rate" and "blood pressure" were studied in the orthostatic tolerance test (pivoting table) and in the bicycle ergometer test. Results were evaluated with variance analysis. Demetrin at dosage levels of 3 x 10 mg/day and 3 x 20 mg/day did not influence cardiovascular function. Occasionally mild sedation was observed.


Dose titration of steroidal and non-steroidal topical anti-inflammatory agents. Although there is a variety of animal models available, neither a single assay system nor the results of the various assays permit absolute protection of relative anti-inflammatory potency. Equally, current clinical dose titration studies, although more reliable, provide only gross estimates of therapeutic potency when conducted in certain clinical situations in a double-blind randomized fashion. This paper delineated clinical means to titrate more  objectively and accurately therapeutic potency in the patient. Moreover, it has submitted considerations as to how to assess therapeutic anti-inflammatory activity in the complex multi-component process of inflammation accompanying most dermatologic diseases that eventually may permit titration of specific anti-inflammatory compounds on certain tissue components of the inflammatory process.


Assessment and treatment of abstinence in the infant of the drug-dependent mother. To accomplish the assessment and treatment of abstinence in the infant of the drug dependent mother, a scoring system has been devised for use as a clinical and research tool. The score monitors the passively addicted infant in a more comprehensive and precise way than has previously been described, and permits us to be more objective in our clinical estimates of the abstinence syndrome. Further, this scoring system has been used by relating it to the dosage schedule of phenobarbital or paregoric as part of an ongoing research project designed to test the comparative usefulness of recommended treatments for neonates with abstinence symptoms. It has been found useful also in following the progression or diminution of symptomatology before, during and after drug therapy. Moreover, the scoring system provides a basis for developing uniform criteria for the assessment and treatment of the neonate of the addicted mother.


Carbenicillin in the first nine months of unrestricted use. A chapter in the natural history of an antibiotic. During the first nine months following its release for general use, carbenicillin was administered to forty-two patients in Hahnemann Hospital. Thirty-nine available records of patients receiving the drug for other than urinary infection were reviewed. Among children, all but one had mucoviscidosis: most were given acceptable therapeutic regimens; survival rate was 80%, and their physicians were favorably impressed. Among adults, underlying disease was usually life threatening; most were inadequately treated; survival rate was 31%, and their physicians were not favorably impressed. The multiplicity of variables precludes objective evaluation of the drug under the circumstances of its use, but one that could have been avoided was the inconsistency of the therapeutic regimen, which in adult patients was too often insufficient. As used, therefore, carbenicillin has not provided the benefits predicted from its earlier controlled evaluation, and the large expenditure of money, particularly by adult patients, has been largely wasted.


Prediction of psychotropic properties of lisuride hydrogen maleate by quantitative pharmaco-electroencephalogram. Based on "quantitative pharmaco-EEG" using computer-analyzed EEG (CEEG) measurements, unknown CNS effects of lisuride hydrogen maleate (LHM) were established. CEEG profiles of LHM in low dosages (less than or equal to 10 mcg) are similar to CNS "inhibitory" compounds, while in higher dosages (25 mcg to 100 mcg) they resemble "psychostimulant" compounds. By measuring the brain function using computer period analysis of cerebral biopotentials, dose-efficacy relations were found (in the range of 25-75 mcg) which suggest the bioavailability of LHM at the CNS level. By comparing the CEEG profiles of LHM with the previously studied compounds, five different clinical uses of LHM were predicted. The pilot trials suggest that LHM may have therapeutic potentials in patients with "aging" and/or organic brain syndromes, and in children with behavioral disturbances.


Clinical pharmacology of SQ 10,996, a potential antidepressive agent. Single oral doses of SQ 10,996 ranging from 500 to 1000 mg (0.34 to 15.3 mg/kg), given once daily for 3 consecutive days to groups of healthy volunteers, were well tolerated. One of three subjects given 1250 mg (17.1 mg/kg) and two of three subjects given 1500 mg (15.6 and 21.4 mg/kg) became drowsy on the second and third days; this symptom disappeared within 24 hrs after cessation of dosage. A short-term, multiple-dose tolerance study was carried out with a formulation of SQ 10,996, the bioavailability of which was comparable to that of the formulation used earlier in the ascending-dose tolerance study. When 200-mg doses were administered every 12, 8, or 6 hrs over a 6-day period, mean steady-state serum concentrations of approximately 4, 7, and 8 mug/ml were attained within 48 hrs; unexpectedly, no subject showed any sign of drowsiness. The half-life for SQ 10,996 in serum, estimated from concentrations in serum after the last dose, was approximately 13 hrs, significantly shorter than the half-life found previously after the administration of single 10-mg doses. These clinical pharmacology studies in healthy volunteers have shown SQ 10,996 to be biologically available and well tolerated. Future studies will test its antidepressive potential in patients.


Serum growth hormone, serum immunoreactive insulin and blood glucose response to oral and intravenous diazepam in man. The effect of diazepam on blood glucose, serum immunoreactive insulin (IRI), and growth hormone (GH) were studied in 10 volunteers who received diazepam in oral doses of 5 and 10 mg, and intravenously 10 mg. They also received placebo and saline treatment. There was a dose-dependent rise in GH after diazepam administration, and the rise was related to the peak plasma level of the drug. A highly significant correlation between the concentrations of serum GH and plasma diazepam was found. During the i.v. and oral administration of 10 mg of diazepam, the peak GH levels, reached in 30 and 60 minutes (19.6+/-2.9 and 15.2+/-3.2ng/ml, respectively), were significantly higher than those during saline and placebo periods (4.3+/-0.8 and 5.9+/-1.1 ng/ml, respectively). There was a tendency to a rise of blood glucose levels, but no significant changes of serum IRI.


Tritiated digoxin: studies in renal disease in human subjects. Digoxin is excreted primarily in the urine as the unchanged glycoside: 60-80% can be recovered from the urine in 7 days after a single intravenous dose in the human subject. Definition of the role of the kidney in digoxin excretion, turnover and metabolism was studied in 57 patients with renal disease, transplant candidates and/or donors and recipients of renal transplants. A single dose of 3H digoxin was given to the subjects, frequent serum samples were obtained and all urine and stools were saved for 7 days. All specimens were extracted with chloroform and digoxin, and its metabolites were separated by column chromatography. Results reveal that the serum T1/2 and the dominant T1/2 of digoxin are prolonged in renal disease in direct proportion to the reduction in creatinine clearance (r = 0.833). The blood urea nitrogen (BUN) is also related to digoxin clearance (r = 0.742). The higher the BUN, the less digoxin excreted in the urine. Anephric patients excrete more digoxin in stool, but this does not compensate for the lack of renal excretion. Transplanted kidneys excrete digoxin in proportion to renal functional capacity, as do patients who have experienced unilateral nephrectomy. Peritoneal or hemodialysis is not effective in removing digoxin from the human subject and may lead to digitalis intoxication if K+ is allowed to fall to critical levels. Digoxin excretion is not volume related, as patients with nephritogenic diabetes insipidus excrete the drug normally with urine volumes of 12 liters a day. Digoxin doses in renal insufficiency should be dictated by knowledge of renal functional ability of the kidneys and after "normal" loading doses, and maintenance doses should be 1/4 to 1/2 those usually administered.


Methods of assessing gingival and periodontal disease: a review. There has been an effort by the dental profession working in the field of gingival and periodontal disease to find a method of recording the extent and degree of pathological change in tissues leading from gingivitis to periodontitis and to measure reversible as well as irreversible changes. It is obvious that some form of index is required and it should have the following well-defined criteria: (1) Simplicity, (2) Accuracy, (3) Quantitativeness, (4) Reproducibility, (5) Speed, (6) Objectivity, and (7) Amenability to statistical analysis. Indices, as well as determining the prevalence of disease in the group under investigation at a given period in time, must also provide information on incidence of disease, i.e. at different periods of time. Indices must also give data that make it possible to verify the nature, severity and aetiology of the disease process and to evaluate therapeutic measures. Indices yield information about the success or failure of control and prevention of disease, affecting the gingivae and the periodontal tissues. A review of methods of assessing and recording gingival and periodontal disease is presented starting from the early investigations of the century. It is shown that subsequently many variations evolved, often with the same inherent difficulties of interpretation or application. Usually the initial stages of inflammation of the gingivae are more difficult to recognize than established disease. Present methods of recording as objectively and as quickly as possible the gingival state in population groups are discussed with emphasis upon the necessity for providing effective preventive measures based upon assessment of schoolchildren.


[Hair growth on pedunoular skin flaps in the guinea pig]. In animal experiments with 12 male guinea pigs skin defects have been covered with one-sided bridge flaps. As opposed to corresponding earlier investigations on free full thickness skin autografts no irreversible and much less reversible damage to the hair follicles could be demonstrated in these skin flaps in the primary postoperative phase. In the later postoperative phase, there was--similar to the free full thickness skin autografts--a synchronization of hair change over several cycles, but on the other hand there was no precipitation of hair change. As a result of these findings it may be assumed that the secondary damage to the hair follicles in the later postoperative phase is not as pronounced in these flaps as in free full thickness skin autografts.


[Acquired epidermolysis bullosa in Crohn's disease]. A case of epidermolysis bullosa acquisita associated with colitis granulomatosa (Crohn's disease) is reported. Although the cutaneous lesions of this unusual disease resemble in their clinical, histological and ultrastructural features to those of the hereditary epidermolysis bullosa dystrophica, the epidermolysis bullosa acquisita has to be regarded as a separate entity among the blistering epidermolysis because of its late manifestation in adults without evidence of hereditary disorders. The deposits of immunoglobulins in the involved skin of the acral areas demonstrated fluorescentmicroscopically are discussed in regard of a possible etiopathogenic relation with the primary irritated intestinal disease.


Enzyme cytochemistry of blood and marrow cells. Diseases of the blood and bone marrow are commonly associated with abnormalities of oxido-reductase and lysosomal enzymes within individual erythrocytes and leucocytes. There are considerable technical difficulties, however, in adapting enzyme histochemical techniques to the study of haemopoietic tissue since individual cells are readily disrupted during processing, show variable enzyme activity according to the stage of maturation, and possess a lipoprotein cytoplasmic membrane which hinders reagent penetration. Cytochemical techniques for the study of oxido-reductase systems are of importance in the study of the neutrophil in infected patients, the erythrocyte in glucose-6-phosphate dehydrogenase deficiency, and the primitive blast cell in acute leukaemia. Lysosomal enzymes are of importance in the study of the neutrophil in infected patients and in the differential diagnosis of acute leukaemia. Some examples of recent studies of these enzyme systems are given to illustrate technical procedures involving cytocentrifugation of cells on to glass slides, adjustment of the osmolality of the reaction mixture, and the study of smeared cells as opposed to cells incubated in suspension.


Retention of Salmonella typhimurium by certain species of fish and shrimp. Marine fish (Mugil cephalus, Fundulus heteroclitus, Trachinotus carolinus), lake shrimp (Penaeus setiferus), and channel catfish (Ictalurus lacustris punctatus) were exposed to Salmonella typhimurium and cultured after 2 hours and at daily intervals for 30 days. Salmonella typhimurium was recovered from all exposed animals after 2 hours and for 30 days in all animals except P setifurus. The organisms could not be recovered from those shrimp after the initial sampling period. Apparent salmonellosis developed in certain specimens of M cephalus and T carolinus 10 to 14 days after exposure to S typhimurium.


Postnatal development of pyrogenic sensitivity in guinea pigs. Despite evidence of thermoregulatory ability from birth, neonates generally are unable to develop fever when challenged with endotoxin. This could be due to their small capacity for heat storage. To test this possibility, the pyrogenicity of S. enteritidis endotoxin (2 mug/kg, iv) was measured at both room (Ta = 25 degrees C) and neutral (Tn = 29-33 degrees C, depending on age) temperatures in 0- to 32-day-old unanesthetized guinea pigs, reared from birth at about 24 degrees C. Control guinea pigs received sterile saline injections in concurrent experiments. Shivering, O2 uptake, and colonic (Tre) and subcutaneous [over the interscapular fat pad (Tbat) and the sacrospinalis muscle (Tsc)] temperatures were recorded continuously for 4 h after injection. Endotoxin generally produced no febrile responses at both ambient temperaturess  rises in animals aged 8 or more days; Tbat increased before the other sites in the 8- and 16-day-old animals, and shivering did not occur; by 32 days of age, however, Tbat no longer increased first, and there was shivering. In Tn significant febrile rises were not evident until 32 days of age; control temperatures, however, were elevated during this exposure as compared to at Ta. These results showed therefore that pyrogenic sensitivity is not apparent in guinea pigs during the first postnatal week; thereafter fever responses are evocable, but their detection may be masked by environmentally produced changes in body temperature. The data also indicated that the site of the heat production underlying, in part, endotoxic fevers gradually shifts from brown fat so skeletal muscle during the first month of life.


Stability in lobar ventilation distribution during change in thoracic configuration. Stability in lobar ventilation was examined in dogs during bilateral electrophrenic respiration (BEPR) and positive pressure-assisted ventilation (PPAV). Bilateral prior ligation of the lower and middle lobe pulmonary arteries monitoring of upper lobe ventilation as alveolar minute ventilation (VA), middle and lower lobe ventilation as dead space (VD), and VD/VT ratio, both calculated by the Bohr equation. As documented by chest films, transverse and anteroposterior thoracic diameters during BEPR decreased below FRC values whereas thoracic cephalocaudal dimension greatly increased. During PPAV, all thoracic dimensions increased. Despite these dissimilar regional chest movements, VA, VD, and VD/VT ratio were comparable between PPAV and BEPR under conditions of matched tidal volume and respiratory frequency. Stability in upper lobe ventilation during BEPR was maintained by caudal displacement despite the compression of the rib cage, as documented by tantalum bronchography. Lobar-interdependence appears to be the mechanism transmitting negative pleural pressure developed by the diaphragm to the upper lobes via lower and middle lobe inflation.


Determination of plasma volume by 113min-labeled transferrin. The binding property of ionic indium (In3+) with plasma transferrin was utilized for determination of plasma volumes (PV) of whole body and individual organs in small animals. Plasma transferrin from a donor rat was labeled with 15-17 muCi 113mIn/ml plasma and injected into the tested rats. PV were determined either by extrapolation to the dilution at time zero (for whole animals) or by calculation of the ratio, organ radiation: radiation of a plasma unit volume (for organs). The reliability of the method for determination of whole-body PV was ascertained by comparing the results obtained with those obtained simultaneously by the Evans blue dilution method. Whole body PV values obtained by the two methods were similar, with a correlation coefficient (r) of 0.997. The short half life of 113mIn enables it to be used with other nuclides which have similar or different energies in the same sample; indium radiation was counted first and after it had disappeared the activity of the other nuclide could be measured.


Maltose transport in Escherichia coli K-12: involvement of the bacteriophage lambda receptor. Mutants affected in lamB, the structural gene for phage lambda receptor, are unable to utilize maltose when it is present at low concentrations (less than or equal 10 muM). During growth in a chemostat at limiting maltose concentrations, the lamB mutants tested were selected against in the presence of the wild-type strain. Transport studies demonstrate that most lamB mutants have deficient maltose transport capacities at low maltose concentrations. When antibodies against purified phage lambda receptor are added to a wild-type strain, transport of maltose at low concentrations is significantly reduced. These results strongly suggest that the phage lambda receptor molecule is involved in maltose transport.


Role of the receptor for bacteriophage lambda in the functioning of the maltose chemoreceptor of Escherichia coli. Chemotaxis towards maltose is specifically defective in many strains of Escherichia coli carrying mutations affecting lamB, the gene coding for the outer membrane receptor for bacteriophage lambda. However, with one exception, the most extreme effect of lamB mutants on the maltose response as determined in the capillary assay is a shift to higher sugar concentrations and a reduction in the number of bacteria accumulated to about 25% of the wild-type level. The severity of the taxis defect is strongly correlated with reduced ability of the cells to take up the maltose present at 1 and 10 muM. Evidence presented here and in the accompanying paper indicates that the lambda receptor is involved in the transport of maltose at these concentrations. The effects of lamB mutations on maltose taxis can be explained by postulating that the high-affinity maltose transport system in which the lambda receptor participates transfers maltose from the surrounding medium across the outer membrane and into the periplasmic space. If the maltose chemoreceptor detects sugar present in the periplasmic space, and not molecules external to the outer membrane, then defective transport of low concentrations of maltose into the periplasm would result in the observed apparent reduction in the sensitivity of the maltose receptor. Thus, the lambda receptor protein would participate in maltose chemorecepton only indirectly through its role in maltose transport.