Computer-assisted diagnosis by a model-free system of direct data analysis. The basis of the method of data analysis presented is, in the case of any diagnostic test, the automatic compilation of separate frequency distributions for each diagnostic classification. The distinction of different test results for different diseases (the correlation for which the tests are used) can thus be quantitatively monitored. This offers opportunities for more specific control of the accuracy of the data base. Measurements of relative frequencies obtained from the frequency distributions of individuals with and without a given disease can serve as a quantitative handle for the selection of the combination of tests, and for adjustments of individual parameters, which will maximize the discrimination. The usual cutoffs are not used. A data-processing system can serve for the direct incorporation of patient chart data (including test results), and for the automation of the analysis described, with pattern recognition or cluster-seeking techniques. The ability of this system of analysis to minimize some of the problems associated with methods utilizing mathematical models is discussed.


Effects of changes in serum osmolarity on bulk flow of fluid into cerebral ventricles and on brain water content. The effects of changes in serum osmolarity on the rate and osmolarity of bulk flow of fluid into the cerebral ventricles and on cortical white and grey matter water content were studied in cats. Bulk flow rates and osmolarities were measured during ventriculocisternal perfusion both before and after intravenous infusion of glucose solutions. Infusions of glucose in concentrations greater than 6% decreased fluid bulk flow rate and its osmolarity. Glucose in concentrations less than 6 percent increased fluid bulk flow rate and decreased its osmolarity. Bulk flow rate and serum osmolarity were found to be linearly related with a coefficient of osmotic flow of minus 0.835 mul/min per mOsm/l. At the extremes of induced serum osmolarities, (290 and 360 mOsm/l) bulk flow rate was either increased by 120 percent or completely inhibited. Effluent osmolarity also increased proportionately to serum osmolarity (0.338 mOsm/l per mOsm/l). When compared to controls, cortical grey and white matter water content increased by 1.9 percent and 2.9 percent, respectively, when the infused glucose concentration was 2.5 percent or less, and decreased by 1.8 percent and 2.9 percent when the concentration was 10 percent or more. The results of these experiments suggest that the increased bulk flow comes from the brain, rather then directly from the blood.


Heterogeneity of mitochondrial DNA from Saccharomyces cerevisiae and genetic information for tRNA. Mitochondrial DNA from wild-type Saccharomyces cerevisiae and from an "extreme" petite mutant were analyzed by hybridization of several tRNAs on DNA fragments of different buoyant density, obtained by sonication and fractionation on a CsCl gradient. The hybridization patterns show that the genes for tRNAser, tRNAphe, tRNAhis, tRNAval, tRNAileu are present on wild-type mitochondrial DNA, while only genes for tRNAser and tRNAhis are present on petite mitochondrial DNA; moreover the hybridization patterns indicate that these genes are not clustered and suggest that more than one gene might exist for tRNAser and tRNAhis.


Primary structure of tRNA Arg II of E. coli B. tRNA Arg II of E. coli has 77 nucleotides. There are eight minor nucleotides including inosine and 2-methyladenosine. Except for a few differences, the structure of tRNA Arg II is very similar to the structure of tRNA Arg I reported by Murao et al.3. The major difference is in the size of dihydrouridine loop. tRNA Arg II does not contain 2-thiocytosine. The unidentified nucleoside X seems to be a different modification other than nucleoside N reported to be present in tRNA Arg I.


Effect of sodium bisulfite modification on the arginine acceptance of E. coli tRNA Arg. Escherichia coli tRNA Arg was treated with sodium bisulfite to convert exposed cytosine residues to uracil. This treatment resulted in the loss of amino acid acceptance of the tRNA Arg with pseudo first-order reaction kinetics. The active and inactive molecules were separated after about 60e active and inactive molecules were separated after about 60 percent inactivation and analyzed for U in various positions by finger-printing of the oligonucleotides produced by nucleases. The results show that C to U base transitions in the dihydrouridine loop and in the CCA terminus have no effect on the aminoacylation of this tRNA. Deamination of a cytosine residue at the second position of the anticodon resulted in the loss of amino acid acceptor activity of arginine transfer RNA.


Physical mapping of the central terminator for transcription on the bacteriophage M13 genome. With the aid of in vitro transcription and translation studies it has been demonstrated that termination of transcription on bacteriophage M13 replicative form DNA occurs at a unique site which is located immediately distal to the 3'-end of gene VIII, the gene which codes for the major capsid protein. The position of this site has been mapped accurately on the enzyme cleavage maps by transcription of restriction fragments of M13 RF DNA. The central termination site was found to be located in restriction fragment Hap-B2 at 450 nucleotides from the 5'-end of its viral strand (0.77 fractional length clockwise from the unique Hind II enzyme cleavage site).


Physical studies of the interaction between the Escherichia coli DNA binding protein and nucleic acids. The interaction of nucleic acid with the Escherichia coli DNA-binding protein has been studied by fluorescence emission spectroscopy and sedimentation velocity analysis. The protein binds to single-strand DNA with an apparent equilibrium dissociation constant of 2 X 10(-9). It binds to the homopolymers poly (dA) and poly (dT) slightly more tightly, but has a larger apparent equilibrium dissociation constant to poly (dC). The protein also binds tightly to ribohomopolymers and to tRNA, but not to duplex DNA. By the use of defined-length oligonucleotides, it has been shown that the protein binds to DNA in a highly cooperative manner. The extent of cooperativity is seen as the difference in binding between an isolated monomeric protein molecule bound to DNA and two or more molecules binding to contiguous sites.


Evidence for tertiary structural RNA-RNA interactions within the protein S4 binding site at the 5'-end of 16S ribosomal RNA of Escherichia coli.+. Evidence is presented for tertiary structural interaction(s) (interactions(s) between two regions of an RNA molecule that are widely separated in the RNA sequence) within the 5'-one third of the 16S ribosomal RNA of Escherichia coli that constitutes the binding site of protein S4. The two main interacting RNA regions were separated by about 120 nucleotides (sections Q to M) of the 16S RNA sequence. A second, smaller gap, of 13 nucleotides, occurred within section C". The two main interacting regions contain about 150 nucleotides (sections H" to Q) and 160 nucleotides (sections M to C"). They are folded back on one another and, especially in the presence of protein S4, are strongly protected against ribonuclease digestion. The intermediate region (sections Q to M), however, is relatively accessible to ribonucleases in the S4-RNP. By partial removal of subfragments from the RNA complex it was possible to localise the two main interacting sites within sections H" - H and sections I" - C". Three main criteria for the specificity of the RNA-RNA interactions were invoked and satisfied. The possibility of other tertiary structural RNA-RNA interactions occurring in other regions of the 16S RNA is discussed. Finally, all the structural information on the S4-RNP is summarised and a tentative model is proposed.


Hybridization of labeled RNA to DNA in agarose gels. Specific DNA restriction endonuclease fragments can be identified after electrophoresis in agarose gels by hybridization in the gel (in situ) to radioactive homologous RNA. RNA-DNA hybrids are detected by autoradiography of the gel. Comparison of band patterns of the autoradiogram and the ethidium bromide stained gel allows the identification of the DNA fragment which is complementary to the RNA probe. The technique is rapid, easy and inexpensive. It is sensitive enough to detect individual genes in a mixture of fragments produced by restriction enzyme digestion of complex cellular DNA. We have used this technique to determine which of the Hin III and Eco R1 fragments of phi80d3ilv+su+7 and E. coli DNAs contain the 5S, 16S and 23S ribosomal RNA (rRNA) genes of E. coli.


The influence of the peptide chain length on the activity of peptidyl-tRNA hydrolase from E. coli. The dependence of the Vmax and Km on the length of the peptide moiety in the peptidyl-tRNA series (Gly)n-Val tRNA, was measured in the system peptidyl-tRNA  hydrolase-peptidyl-tRNA. It was found that the Km value decreases from 7.2 X 10-7 M for Gly-Val-tRNA to 4.6 X 10-7 M FOR (Gly)2-Val-tRNA and to 1.7 X 10-7M for (Gly)3-Val-tRNA; further increase of the peptide chain is not followed by decrease of the Km. The Vmax values are 5.7 pmole/min/EU for Gly-Val-tRNA and 42 pmole/min/EU for (Gly)3-Val-tRNA. The enzyme activity is inhibited competitively by uncharged tRNA with a KI value of about 10-5M. The significance of these results described in this paper, in relation to the fact that peptides and peptide esters do not inhibit the enzyme activity, and in relation to the proposed physiological role of the enzyme, is discussed.


[Effect of antibiotics on hemolysin production by bacteria causing mastitis in cattle]. Effect of six antibiotics on the hemolysin formation by 320 strains of Staphylococcus (including 112 S. aureus) and 100 strains of S. agalactiae isolated from cow udders, have been determined. Tests were performed on plates containing 5% horse red cells, but the hemolytic strains were tested on blood agar plates supplemented with 5% sheep, rabbit and calf cells in addition. One strain of S. aureus produced the rings of hemolysis around the zones of growth inhibition on the media containing the discs of penicillin, erythromycin, oxyterramycin and chloramphenicol. Another strain of S. agalactiae produced similar rings, but only on the medium supplemented with penicillin. On the calf blood agar plate, S. aureus produced the hemolytic rings only in association with streptomycin and neomycin. After five times of propagation on the nutrient agar, the hemolytic phenomenon was not reproducible. It may be suggested that in cow milk there are some inhibitors for hemolysin formation by Staphylococcus and S. agalactiae.


[Course of beta-hemolytic E. coli carrier state in piglets during weaning]. The investigations were carried out on 22 sows and 44 piglets (one sow and 2 piglets from each litter). The pigs were divided into 4 groups according to the different additions to their diet during weaning which might have an effect on the carrier state and dissemination of beta-haemolytic E. coli. Group I was given a feed concentrate and separated milk, group II - a feed concentrate and sour milk, group III - a feed concentrate and milk with addition of garlic (200 mg/kg of body weight), group IV - a feed concentrate and milk with addition of vitamin C (20 mg/kg of body weight). The beta-haemolytic E. coli carrier state was examined before starting supplemental feeding (3 times between the 21st and 30th day of life) and after weaning (3 times between the 49th and 56th day of life). The carrier state under discussion was found in 27.2% of sows and in 25% of piglets at the beginning of supplemental feeding, and in 40.1% of sows and 36.3% of piglets during weaning, which means an increase of the number of carriers both among the sows and piglets at that time. Twenty-two strains of E. coli isolated from the carriers belonged to serotypes responsible for colibacteriosis of pigs (O149:K91, K88a, c, O8:K87, K88a, c, O139:K82); 23 strains belonging to E. coli groups O6, O8 and O116 were devoid of K and H antigens, 9 strains possessed a double O antigen (O9, O60: :H-, O117, O120: :H-, O102, O25: :H21). An inhibitory effect of the addition of sour milk to the feed on the intensity of the carrier state of beta-haemolytic E. coli was observed in piglets kept on that diet. The pathogenic strains of E. coli were observed more often in the carrier-piglets (60%), and in the sows predominated the strains of beta-haemolytic E. coli devoid of the K antigens.


Candida species and crop mycosis in broiler chickens. A mycological survey of the crops of approximately 100 healthy birds from each of 6 grow-out operations revealed that the incidence of Candida in the crops ranged from 17.4 to 51.5% with a mean value of 32.3%. The population of Candida in the crops of birds found positive was of low magnitude in the majority of the chickens examined. Of the 573 birds examined in this study less than 1% exhibited visible lesions attributable to Candida. C. albicans comprised 95% of the isolates while C. ravautii, C. salmonicola, C. gulliermondi, C. papapsilosis, C. catenulata and C. brumptii comprised the remainder. The incidence and number of Candida in the crop was related apparently to management practices on the farm. The crops from four field outbreaks of crop mycosis were also studied. Three of the four cases of crop mycosis were characterized by multiple strains of C. albicans in the crop. In one case, C. parapsilosis also was isolated from the crop.


Interaction of T-2 toxin with Salmonella infections of chickens. A significant (P less than 0.05) interaction resulting in increased mortality occurred in chickens fed T-2 toxin (16 mug./g. of diet) and infected with either Salmonella worthington, S. thompson, S. derby, or S. typhimurium var. copenhagen, all species that cause paratyphoid. No interaction on growth rate or relative size of the bursa of Fabricus occurred, although T-2 toxin alone caused a significant (P less than 0.05) regression of that organ. The spleen size relative to the body weight was decreased (P less than 0.05) by T-2 toxin and increased (P less than 0.05) by the Salmonella infections. Interactions were observed on spleen size between the toxin and S. thompson (P less than 0.05) and S. derby (P less than 0.10). Total serum proteins were not affected by T-2 toxin or Salmonella infections. Agglutinins were formed in response to the infections, but the titers were unaltered by T-2 toxin.


Feprazone compared with indomethacin in the management of rheumatoid arthritis. A double-blind cross-over trial of feprazone 450 mg daily and indomethacin 75 mg daily was carried out in fourteen patients with rheumatoid arthritis. The analgesic and anti-inflammatory activity was indistinguishable from that of indomethacin under the conditions of the trial. Seven patients expressed a preference for feprazone and four for indomethacin. Feprazone appeared to be well tolerated and free from serious side-effects. These results suggest that feprazone will be a useful drug in the management of rheumatoid arthritis.


Removal of unwanted proteins from cell extracts by means of antiserum. The efficiency of removal of soluble proteins in cell-free bacterial extracts by means of antiserum from rabbits immunized with similar extracts was measured. Precipitation followed by Sephadex gel-chromatography was used. Up to 80% (exceptionally 90%) removal could be obtained. The method might be applied to enrichment for "foreign" cell-extract components, for example, viral products in virus infected cells. Tests for the specificity of the method are also presented.


[Production of the methyl ether of 8,14-seco-delta,3,5,(10),9(11)-estratetraendiol-3,17beta-one-14 using a culture of Saccharomyces cerevisiae BKMU-488]. The culture Saccharomyces cerevisiae BKMU-499 was selected from 100 strains of different Saccharomyces species to obtain an optically active methyl ester 8,14-seco-delta1,3,5(10),9(11)-estratetraendiol-3-17beta-on-14. Various conditions of the transformation were examined. The yield of this keto-alcohol depended on the following factors: level of aeration, method of steroid administration, physiological state of the culture, and composition of the cultivation medium. The optimal yeild of the optically active product was 84%.


Effects of guanosine tetraphosphate on cell-free synthesis of Escherichia coli ribosomal RNA and other gene products. A cell-free system derived from E. coli is described in which mature-sized 16S and 23S ribosomal RNAs (rRNA) are synthesized at a high relative rate, comprising 17-25% of the total transcription. The addition of guanosine tetraphosphate (ppGpp) to this system results in up to a 5-fold selective inhibition of rRNA accumulation. This effect is exerted at the level of synthesis rather than degradation. It is concluded that ppGpp, which is produced in large amounts by E. coli during amino-acid deprivation, could mediate the decrease in rRNA synthesis that accompanies such deprivation. The expression of other genes has also been investigated. No selective reduction of transfer RNA synthesis by ppGpp is observed. The trp and lac operons are found to be stimulated at the transcriptional level by the presence of this nucleotide. It is hypothesized that ppGpp interacts with the RNA polymerase in such a manner as to alter the affinity of the enzyme for promoters in an operon-specific fashion.


Very stable prokaryotic messenger RNA in chromosomeless Escherichia coli minicells. E. coli minicells lack DNA, yet they make protein, the synthesis of which is sensitive to chloramphenicol but insensitive to rifamycin. This protein is coded for by very stable cellular mRNA with an estimated half-life of 40-80 min. In an R factor-containing minicell, two very different species of mRNA are observed: (i) R factor-specific mRNA with a short half-life whose synthesis is rifamycin-sensitive and (ii) cellular mRNA with a long half-life whose synthesis is rifamycin-insensitive. These findings indicate that minicells contain normal degradative mechanisms for mRNA and point out the existence of a unique class of very stable cellular mRNA. Greater than 80% of the rifamycin-insensitive protein synthesized goes into the outer minicell membrane. Relatively stable mRNA, half-life 5.5-11.5 min, for outer membrane protein in whole cells has been reported [Hirashima et al. (1973) J. Mol. Biol. 79, 373-389]. The stability of minicell mRNA is significantly greater. This and other observations suggest that there are two functional species of mRNA for outer membrane protein perhaps in different sites in the cell. Furthermore, these studies suggest that a class of cellular proteins is synthesized in bacteria without concomitant transcription and in the absence of association with chromosomal DNA.


Bidirectional replication of plasmid R6K DNA in Escherichia coli; correspondence between origin of replication and position of single-strand break in relaxed complex. Replicating molecules of plasmid R6K DNA have been purified as covalently closed circular DNA forms and analyzed in the electron microscopy after cleavage with the EcoRI restriction endonuclease. It has been determined that in most cases replication proceeds bidirectionally from an origin whose position is indistinguishable from the site of the single-strand break (nick) in the open circular DNA form of the relaxation complex of R6K DNA and protein. Evidence is presented for the existence of a unique replication terminus asymmetrically placed approximately 20% of genome size from the origin. The positions of the replication forks in a majority of the molecules indicate that replication proceeds sequentially from the fixed origin first in one direction to the terminus and then progresses from the origin in the other direction.


Presence of polyriboadenylate sequences in pulse-labeled RNA of Escherichia coli. Pulse-labeled RNA isolated from E. coli cells grown on limiting phosphate medium and phosphate-containing medium was analyzed by oligo(dT)-cellulose chromatography and by Millipore binding assay for polyriboadenylate-containing RNA. Whereas poly(A)-containing RNA amounted to as much as 15% of the total pulse-labeled RNA from cells grown on limiting phosphate medium, pulse-labeled RNA from cells grown on phosphate medium gave values around 1.5%. Steady-state labeled RNA from cells grown on limiting phosphate medium contained 1.2% poly(A) RNA. The addition of poly(A) sequences appears to be post-transcriptional. These results strongly favor the view that bacterial mRNAs may contain poly(A) stretches.


Interaction of enzyme I of the phosphoenolpyruvate:sugar phosphotransferase system with adenylate cyclase of Escherichia coli. Transient repression by glucose of induced enzyme synthesis involves lowering of intracellular cAMP levels. This glucose effect is partially explained by a glucose inhibition of adenylate cyclase [EC 4.6.1.1; ATP pyrophosphate-lyase(cyclizing)]. Since the phosphoenolpyruvate:sugar phosphotransferase system has been implicated in repression phenomena, an investigation was made of adenylate cyclase activity in mutants of that transport system. The results suggest that glucose phosphorylation is not necessary for inhibition of adenylate cyclase since an HPr mutant retained sensitivity to glucose inhibition. The results also suggest that adenylate cyclase activity requires the presence of Enzyme I in a phosphorylated form and that adenylate cyclase activity may be regulated by a phosphorylation-dephosphorylation mechanism.


Binding of ribosomal protein S1 of Escherichia coli to the 3' end of 16S rRNA. Ribosomal protein S1 reversibly binds the 49-nucleotide fragment that is cleaved from the 3' end of 16S rRNA in ribosomes by colicin E3. The fragment has secondary structure in the form of a hairpin loop. At the base of the stem is a sequence (A-C-C-U-C-C) thought to be involved in the base pairing with complementary sequences in mRNA during the initiation of protein synthesis. The role of S1 may be to stabilize this region of the fragment in an open conformation to allow for base pairing to mRNA. This model is supported by the observation that S1 binds specifically to this region of the fragment. In addition, aurin tricarboxylic acid, an inhibitor of protein synthesis, reverses this effect by disrupting the S1-RNA complex. These results can explain why S1 is an essential component of the ribosome for translation of natural mRNA and why aurin tricarboxylic acid blocks initiation.


Photoincorporation of puromycin and N-(ethyl-2-diazomalonyl)puromycin into Escherichia coli ribosomes. [3H]Puromycin and N-(ethyl-2-diazomalonyl)[3H]puromycin are incorporated into E. coli ribosomes on irradiation at 253.7 nm. Both compounds incorporate into both protein and nucleic acid. Two-dimensional gel electrophoresis of ribosomal protein shows that L23 is the major protein labeled by puromycin. Although incorporation is clearly a complex process, evidence is presented that L23 is labeled via an affinity labeling process, thus placing L23 at the aminoacyl-tRNA receptor (A) site. N-(ethyl-2-diazomalonyl)puromycin is a ribosomal ligand, as shown by its inhibition of two ribosomal assays, but it is not a good puromycin analog, and it is unclear whether its incorporation, which proceeds via both carbene-dependent and carbene-independent processes, results from affinity labeling.


Role of magnesium in Escherichia coli alkaline phosphatase. Alkaline phosphatase of E. coli, isolated by procedures which do not alter its intrinsic metal content, contains 1.3 +/- 0.3 g-atom of magnesium and 4.0 +/- 0.2 g-atom of zinc per molecule of molecular weight 89,000. Magnesium, the role of which has been unappreciated, significantly affects the function and structure of alkaline phosphatase containing either 2 or 4 g-atom of zinc per mole. Magnesium does not activate the apoenzyme but increases the activity of the enzyme containing 2 g-atom of zinc 4.4-fold and that of the enzyme containing 4 g-atom 1.2-fold. The results obtained with enzyme in which cobalt is substituted for zinc are analogous. Moreover, the absorption and electron paramagnetic resonance spectra of cobalt phosphatases reveal the effects of magnesium on cobalt coordination geometry. Addition of magnesium changes the spectral characteristics of the apoenzyme reconstituted with 2 g-atom of cobalt from predominantly octahedral to 4- or 5-coordinate geometry. These two classes of cobalt binding sites have been associated with catalysis and structure stabilization, respectively. Therefore, magnesium controls the occupancy of the catalytic and structural binding sites and modulates the resultant enzymatic activity. Hydrogen-tritium exchange was employed to determine the effects of magnesium on the conformational stability of phosphatase. Magnesium stabilizes the dynamic structural properties, both of apophosphatase and of enzyme containing 2 g-atom of zinc, which is further stabilized by 2 more zinc atoms. The role of magnesium and other metal ions in regulatory processes, only now beginning to be explored fully, will likely emerge as an important avenue for achievement of regulatory effects in metalloenzymes.


Distinct penicillin binding proteins involved in the division, elongation, and shape of Escherichia coli K12. The varied effects of beta-lactam antibiotics on cell division, cell elongation, and cell shape in E. coli are shown to be due to the presence of three essential penicillin binding proteins with distinct roles in these three processes. (A) Cell shape: beta-Lactams that specifically result in the production of ovoid cells bind to penicillin binding protein 2 (molecular weight 66,000). A mutant has been isolated that fails to bind beta-lactams to protein 2, and that grows as round cells. (B) Cell division: beta-Lactams that specifically inhibit cell division bind preferentially to penicillin binding protein 3 (molecular weight 60,000). A temperature-sensitive cell division mutant has been shown to have a thermolabile protein 3. (C) Cell elongation: One beta-lactam that preferentially inhibits cell elongation and causes cell lysis binds preferentially to binding protein 1 (molecular weight 91,000). Evidence is presented that penicillin bulge formation is due to the inhibition of proteins 2 and 3 in the absence of inhibition of protein 1.


Sigma cycle during in vitro transcription: demonstration by nanosecond fluorescence depolarization spectroscopy. Studies of RNA chain initiation have suggested that the sigma subunit of Escherichia coli RNA polymerase (RNA nucleotidyltransferase; nucleosidetriphosphate: RNA nucleotidyltransferase; EC 2.7.7.6) is released from the enzyme-template complex during transcription and may be reused by another core polymerase. Nanosecond fluorescence depolarization spectroscopy was used to follow the sigma cycle. Isolated sigma subunit labeled with the fluorescent probe dansyl (DNS) chloride bound stoichiometrically to core polymerase and stimulated transcription of phage T7 DNA to the same extent as did unlabeled sigma. DNS-sigma showed an exponential fluorescence anisotropy decay corresponding to a rotational correlation time of about 100 nsec. This value was unaffected by addition of T7 DNA, but increased about 6-fold when core polymerase was added, and increased further when T7 DNA was added. Such increases are expected for the formation of molecular complexes. Using the anisotropy decays for free DNS-sigma and DNS-sigma-core enzyme bound to T7 DNA, we calculated theoretical decay curves for various mixtures of free and bound sigma. Comparison of the observed anisotropy decay with the calculated curves indicated that about 55% of DNA-sigma was released from the enzyme-T7 DNA complex in the presence of four nucleoside triphosphates under low salt conditions. Sigma release did not occur if rifampicin was added prior to addition of four nucleoside triphosphates or if only three nucleoside triphosphates were present. After sigma was released, addition of core polymerase with rifampicin reduced the free sigma to less than 15%, indicating that the released sigma was accessible to the added core enzyme. Thus these studies have provided physical evidence for the sigma cycle during in vitro transcription.


Visualization of a novel junction in bacteriophage lambda DNA. At early times after infection of a recA derivative of Escherichia coli with lambdab221c126red270a42 phage, a low but significant proportion of intracellular lambda molecules show a novel junction. These junctions are also present, although in reduced numbers, in a lysate obtained at late times after infection of a recA+ host with lambdacIIcIII phage. Fine structure and denaturation mapping analyses showed that these junctions occur at homologous positions and that they are compatible with the occurrence of a cross-strand exchange between lambda DNA duplexes similar to the type proposed in most molecular models for genetic recombination. However, the results are also consistent with the structures expected if a replicating growing point undergoes branch migration.


Dissociation of two polypeptide chains from yeast RNA polymerase A. Yeast RNA polymerase A (RNA nucleotidyltransferase; nucleosidetriphosphate:RNA nucleotidyltransferase; EC 2.7.7.6) can be converted to a new form of enzyme, called RNA polymerase A*, which is lacking two polypeptide chains of 48,000 and 37,000 daltons. Apart from these two missing polypeptides the subunit structures of RNA polymerases A and A* are indistinguishable. RNA polymerase A* differs from the complete enzyme in its electrophoretic and chromatographic behavior, template requirements, and alpha-amanitin sensitivity. RNA polymerase A* transcribes the alternated copolymer d(A-T)n with the same efficiency as RNA polymerase A but its specific activity is greatly reduced with native calf thymus DNA as template. The transcription of a variety of synthetic templates is also altered by removal of the two polypeptide chains. RNA polymerase A* is inhibited by high concentrations of alpha-amanitin (500 mug/ml), whereas RNA polymerase A is comparatively less sensitive to the toxic peptide. The data are discussed in terms of possible roles of the two dissociable polypeptides.


Amino acids are not all initially attached to the same position on transfer RNA molecules. Escherichia coli tRNA has been modified by replacement of the 3'-terminal AMP with either 3'-amino-3'-deoxy AMP of 2'-amino-2'-deoxy AMP. These tRNA analogs have enabled us to determine the initial site of enzyme-catalyzed aminoacylation of different tRNAs by the formation of aminoacyl-tRNA molecules in which the amino acid is linked to the 3'-terminal ribose through a stable amide bond. The tRNA species specific for glutamic acid, glutamine, leucine, phenylalanine, tyrosine, and valine are all aminoacylated on the 2'-hydroxyl group. The tRNA species specific for alanine, asparagine, aspartic acid, glycine, histidine, lysine, and threonine are aminoacylated on the 3'-hydroxyl group. The amino acids arginine, isoleucine, methionine, proline, serine, and tryptophan form stable amide bonds with both amino tRNA analogs. This might suggest that the synthetases for these amino acids can acylate both the 2'- and 3'-hydroxyl groups, but it is more likely that these enzymes can acylate both hydroxyl and amino groups at either the 2' or 3'-position of the tRNA. These results clearly illustrate a fundamental heterogeneity which is apparent in the mechanism of action of aminoacyl-tRNA synthetases.


Site of aminoacylation of tRNAs from Escherichia coli with respect to the 2'- or 3'-hydroxyl group of the terminal adenosine. A method is presented by which the site of primary attachment of the amino acids with respect to the 2'- or 3'-hydroxyl group of the terminal adenosine of E. coli tRNAs can be determined. It is found that the aminoacyl-tRNA synthetases (EC 6.1.1.-) with specificity for Arg, Asn, Ile, Leu, Met, Phe, Thr, Trp, and Val attach the amino acid to the 2'-position; those with specificity for Gly, His, Lys, and Ser attach the amino acid to the 3'-position; and that Tyr and Cys can be enzymatically attached to both the 2'- and 3'-positions. Together with previous experiments on yeast aminoacyl-tRNA synthetases, it is now shown that the specificity for one particular hydroxyl group is preserved during the evolution from prokaryotic to eukaryotic systems.


Reconstitution of the Escherichia coli pyruvate dehydrogenase complex. The binding of pyruvate dehydrogenase and dihydrolipoyl dehydrogenase (flavoprotein) to dihydrolipoyl transacetylase, the core enzyme of the E. coli pyruvate dehydrogenase complex [EC 1.2.4.1:pyruvate:lipoate oxidoreductase (decaryboxylating and acceptor-acetylating)], has been studied using sedimentation equilibrium analysis and radioactive enzymes in conjunction with gel filtration chromatography. The results show that the transacetylase, which consists of 24 apparently identical polypeptide chains organized into a cube-like structure, has the potential to bind 24 pyruvate dehydrogenase dimers in the absence of flavoprotein and 24 flavoprotein dimers in the absence of pyruvate dehydrogenase. The results of reconstitution experiments, utilizing binding and activity measurements, indicate that the transacetylase can accommodate a total of only about 12 pyruvate dehydrogenase dimers and six flavoprotein dimers and that this stoichiometry, which is the same as that of the native pyruvate dehydrogenase complex, produces maximum activity. It appears that steric hindrance between the relatively bulky pyruvate dehydrogenase and flavoprotein molecules prevents the transacetylase from binding 24 molecules of each ligand. A structural model for the native and reconstituted pyruvate dehydrogenase complexes is proposed in which the 12 pyruvate dehydrogenase dimers are distributed symmetrically on the 12 edges of the transacetylase cube and the six flavoprotein dimers are distributed in the six faces of the cube.


Cleavage of adenovirus messenger RNA and of 28S and 18S ribosomal RNA by RNase III. Escherichia coli ribonuclease III cleaves adenovirus messenger RNA and mammalian 28S and 18S ribosomal RNA. Fragmentation is not random, but in each case a specific collection of products is generated. This points to the potential use of the enzyme as a tool for specific fragmentation of RNA. Cleavage by RNase III abolishes the capability of adenovirus messenger RNA to direct cell-free synthesis of virus polypeptides.


Mutagenic and cytotoxic activity of benzol[a]pyrene 4,5-, 7,8-, and 9,10-oxides and the six corresponding phenols. The benzo[a]pyrene 4,5-, 7,8-, and 9,10-oxides and the six corresponding phenols (4-, 5-, 7-, 8-, 9-, and 10-hydroxybenzo[a]pyrene) have been tested for mutagenic and cytotoxic activity in bacteria and in a mammalian cell culture system. Benzo[a]pyrene 4,5-oxide (K-region) was highly mutagenic in two histidine-dependent strains (TA1537 and TA1538) of Salmonella typhimurium which detect frameshift mutagens. In contrast, benzo[a]pyrene 7,8- and 9,10-oxides were less than 1% as mutagenic as the 4,5-oxide. Benzo[a]pyrene 7,8- and 9,10-oxides were unstable in aqueous media, whereas the 4,5-oxide was stable for several hours. This difference in stability could not account for the different mutagenic activities of the three arene oxides. The benzo[a]pyrene oxides were inactive in a strain (TA1535) that is reverted by base pair mutagens such as N-methyl-N'-nitro-N-nitrosoguanidine or in a strain (TA1536) that detects framshift mutagens similar to the acridine half-mustard ICR-191. Benzo-[a]-pyrene and the six phenols were all stable in aqueous media, but they had little or no mutagenic activity in any of the four Salmonella strains. Conversion of 8-azaguanine-sensitive Chinese hamster V79 cells to 8-azaguanine-resistant variants was increased by benzo[a]pyrene 4,5-oxide, whereas the 9,10-oxide was considerably less active. Benzo[a]pyrene and the other derivatives had little or no effect. Benzo[a]yrene 4,5-oxide was more cytotoxic to the Chinese hamster V79 cells than the 7,8- and 9,10-oxides, while 8-hydroxybenzo[a]pyrene was the most cytotoxic of the six phenols.


Leukocyte migration inhibitory factor (LMIF) induced by concanavalin A: standardized microassay for production in vitro. A standardized microculture system has been developed to assess the ability of lymphocytes to secrete leukocyte migration inhibitory factor (LMIF) in response to the nonspecific mitogen concanavalin(Con A). LMIF-rich supernates collected from stimulated lymphocytes cultured in plastic microtiter plates are assayed by pulse exposure of purified human granulocytes and inhibition of their migration in agarose medium. LMIF activity in this system is suppressed by the protein synthesis inhibitor puromycin, but not by inhibition of lymphocyte proliferation by irradiation. It is demonstrated that normal lymphocytes stimulated with mitogen elaborate LMIF activity, while lymphocytes from malignant lymphoma patients are frequently unable to produce it. Thus, mitogen-induced mediator production may be a useful parameter in further characterization of primary and secondary immunodeficiencies.


Recovery of infectious proviral DNA from mammalian cells infected with respiratory syncytial virus. The DNA fraction from a line of bovine embryonic kidney cells originally exposed as primary cultures several months earlier to a temperature-sensitive (ts) mutant of respiratory syncytial (RS) virus could be used to transfect human HEp-2 cells with the production of infectious RS virus. The DNA donor cells, designated BEK/RS ts, retained their healthy fibroblastic appearance during continuous cultivation at a temperature (39 degrees) restrictive for growth of the original infecting mutant and showed no evidence for RS virus replication or viral antigen synthesis when directly examined for these activities by conventional methods. The infectious property of the DNA from BEK/RS ts cells was abolished by exposure of the nucleic acid preparation to DNase (but not RNase) or by pretreatment of recipient HEp-2 cells with actinomycin D or mitomycin C. The latter drug treatments substantially enhanced the replication of infecting wild-type RS virus in HEp-2 cells. Viral isolates derived from the progeny of a DNA transfection included clones possessing several genetic markers of the RS ts mutant originally used to infect BEK/RS ts cells and other virus clones that appeared to be either hybrid or wild-type for phenotypic properties such as their temperature sensitivity. An infectious proviral DNA was also detected in a line of virogenic HEp-2 cells (HEp-2/RS) persistently infected with respiratory syncytial virus after exposure to the wild-type strain 2 years earlier.


Transient response to chemotactic stimuli in Escherichia coli. We have followed by eye and with the tracking microscope the rotational behavior of E. coli tethered to coverslips by their flagella. The cells change their directions of rotation at random, on the average about once a second. When an attractant is added or a repellent is subtracted, they spin clockwise (as viewed through the coverslip, i.e., along the flagellum toward the body) for many seconds, then counter-clockwise for many seconds, and then gradually resume their normal mode of behavior. The time interval between the onset of the stimulus and the clockwise to counter-clockwise transitiion is a linear function of the change in receptor occupancy. The cells adapt slowly at a constant rate to the addition of an attractant or the subtraction of a repellent. They adapt rapidly to the subtraction of an attractant or the addition of a repellent. Responses to mixed stimuli can be analyzed in terms of one equivalent stimulus.


Sequential processing of precursor tRNA molecules in Escherichia coli. In a temperature-sensitive mutant of E. coli defective in tRNA biosynthesis, many tRNA precursors, including monomeric and multimeric forms, accumulate. Some of the multimeric precursors contain three or more tRNA sequences within a molecule. These large precursors were cleaved by cell extracts first into intermediate size pieces which were subsequently processed by RNase P. On the basis of heat stability of mutant cell extracts, the endonuclease responsible for the initial cleavage appears to be distinct from RNase P and is designated RNase O. One of the monomeric precursors was shown to be processed first by RNase P and the product subsequently cleaved further into a smaller molecule. The nuclease responsible for this second cleavage also appears to be distinct from RNase P and is designated RNase Q. The functions of these nucleases are sequential in the trimming process with respect to that of RNase P; RNase O works prior to RNase P and RNase Q after RNase P but in both cases, not vice versa.


Conformational change and cooperativity in actin filaments free of tropomyosin. The decrease in amplitude of the electron spin resonance spectrum of the cysteine-bound spin-label, 3-(maleimidomethyl)-2,2,5,5-tetramethyl-1-pyrrolidinoxyl, brought about by the magnetic interaction with tightly bound manganous ion, was used as a probe of conformational change in actin on binding myosin. The magnitude of this "spin--spin" interaction first decreased then increased on increasing saturation of the actin filament with heavy meromyosin subfragment-1. That the "spin--spin" interaction occurred between spins of adjacent monomers was demonstrated by the observation that the change in magnitude of the "spin--spin" interaction was maintained on binding of heavy meromyosin subfragment-1 to copolymers in which actin monomers containing both manganous ion and spin label were diluted 7-fold with native actin monomers. These data provide evidence for a conformational change in actin on interacting with heavy meromyosin subfragment-1. Further, the fact that not only the magnitude but also the sense of the change in the "spin--spin" interaction is a function of increasing saturation with heavy meromyosin subfragment-1 indicates that the monomers of the actin filament are capable of cooperative interaction in the absence of tropomyosin.


In vitro construction of bacteriophage lambda carrying segments of the Escherichia coli chromosome: selection of hybrids containing the gene for DNA ligase. DNA from lambdagt-lambdaB bacteriophage was cleaved with EcoRI endonuclease and fragments from EcoRI-digested E. coli DNA were inserted. This DNA was used to infect E. coli, and phages containing the gene for DNA ligase were isolated by genetic selection. Two different hybrids were found with the same E. coli segment inserted in opposite orientations. Both hybrids produced similar levels of ligase as measured in crude extracts of infected cells.


Kinetics of action of pepsin on fluorescent peptide substrates. Oligopeptide substrates of porcine pepsin (E) of the type A-Phe-Phe-B (S) that are cleaved solely at the Phe-Phe bond under the conditions of these studies, and bearing an amino-terminal fluorescent probe group (mansyl or dansyl), have been used for stopped-flow measurements of the rate of formation of the A-Phe product. These experiments were conducted under conditions of [E] greater than [S], and the kinetic data were compared with those obtained under conditions of [S] greater than [E] for the formation of the Phe-B product (the same in all cases). The results for substrates with A = mansyl-Gly, mansyl-Gly-Gly, and dansyl-Gly-Gly support the conclusion that the rate-limiting step in the over-all catalytic process is associated with the scission of the Phe-Phe bond in the first detectables ES complex. Although the rate of this step varies widely with the nature of the A portion of A-Phe-Phe-B, the magnitude of the dissociation constant of ES is relatively invariant. This supports the view that, in the cleavage of oligopeptide substrates by pepsin, secondary enzyme--substrate interactions may cause conformational changes at the catalytic site, and that a portion of the total binding energy may be used for the attainment of the transition state in the bond-breaking step. With substrates that are hydrolyzed extremely rapidly (A = dansyl-Gly-Ala, dansyl-Ala-Ala), the rate of formation of the A-Phe product appears to be faster than the steady-state rate, suggesting that an additional step has become kinetically significant in the over-all process. This step may be associated with the return of the conformation of the active site to its original state.


Characterization of a novel, low-molecular-weight DNA-binding protein from Escherichia coli. A low-molecular-weight (7000), heat-stable protein--HU--that stimulates transcription of bacteriophage lambda DNA by E. coli RNA polymerase was purified from E. coli extracts using affinity chromatography on DNA-cellulose. HU binds to native DNA, resulting in an apparent thickening of the DNA chains as revealed by electron microscopy. Contrary to DNA unwinding proteins, it causes no destabilization of the double helix. HU differs from previously described transcription factors (H1, D, etc.) and from the low-molecular-weight omega subunit of the RNA polymerase. By its amino-acid composition and characteristics, HU displays an interesting resemblance to some eukaryotic histones, such as H2B and H1.


Histidine regulation in Salmonella typhimurium: an activator attenuator model of gene regulation. An activator-attenuator model of positive control, a s opposed to the classic repressor-operator model of negative control, is proposed for the major operon-specific mechanism governing expression of the histidine gene cluster of Salmonella typhimurium. Evidence for this mechanism is derived from experiments performed with a coupled in vitro transcription-translation system, as well as with a minimal in vitro transcription system [Kasai, T. (1974) Nature 249, 523--527]. The product (G enzyme, or N-1-[5'-phosphoribosyl]adenosine triphosphate:pyrophosphate phosphoribosyltransferase; EC 2.4.2.17) of the first structural gene (hisG) of the histidine operon is not involved in the positive control mechanism. However, a possible role for G enzyme as an accessory negative control element interacting at the attenuator can be accommodated in our model. The operon-specific mechanism works in conjunction with an independent mechanism involving guanosine 5'-diphosphate 3'-diphosphate (ppGpp) which appears to be a positive effector involved in regulating amino-acid-producing systems, in general [Stephens, J.C., Artz, S.W. & Ames, B.N. (1975) Proc. Nat. Acad. Sci. USA, in press].


Translocation of a plasmid DNA sequence which mediates ampicillin resistance: molecular nature and specificity of insertion. A series of recombinant plasmids was generated in Escherichia coli in which the TEM beta-lactamase translocon (TnA) was inserted into the small plasmid RSF1010. RSF1010 is a 5.5 X 10(6) dalton nonconjugative plasmid which confers resistance to streptomycin and sulfonamide. The recombinant plasmids can be classified into three clearly defined phenotypic groups. Group I is ampicillin-, streptomycin- and sulfonamide-resistant. Group II is ampicillin- and sulfonamide-resistant but has lost streptomycin resistance. Group III is ampicillin-resistant but is sensitive to sulfonamide and shows a simultaneous 30-fold reduction in the minimal inhibitory concentration of streptomycin. It was possible to map the site of insertion of TnA within RSF1010 by electron microscope studies of DNA heteroduplexes formed between RSF1010 and recombinant plasmids. Insertions of TnA occur at, at least, 12 distinct sites in a region corresponding to one-third of the RSF1010 DNA molecule. Those insertions giving rise to particular phenotypes are clustered. Insertions of TnA-like insertion sequences (IS) appear to give rise to strongly polar mutations.


Transformation of Pseudomonas putida and Escherichia coli with plasmid-linked drug-resistance factor DNA. Conditions optimal for the transformation of Pseudomonas putida and E. coli with a drug-resistance factor (RP 1) DNA, which specifies resistance to carbenicillin, tetracycline, kanamycin, and neomycin, are described. The transformants retain all the fertility, incompatibility, and drug-resistance characteristics present in the parent. Covalently-closed circular molecules of almost identical contour lengths have been isolated from the parent and the transformants. The frequency of transformation is drastically reduced by treatment of RP 1 DNA with DNase and by denaturation or sonication. Shearing of RP 1 DNA in vitro and their subsequent introduction in P. putida cells, by transformation, produces transformants that exhibit a wide range of drug-resistant phenotypes, including those which are resistant to neomycin but sensitive to kanamycin. Isolation of such neomycin-resistant but kanamycin-sensitive transformants indicates that there might be two separate mechanisms specified by RP 1 for resistance to the two antibiotics.


An endotoxin-induced serum factor that causes necrosis of tumors. In studying "hemorrhagic necrosis" of tumors produced by endotoxin, it was found that the serum of bacillus Calmette--Guerin (BCG)-infected mice treated with endotoxin contains a substance (tumor necrosis factor; TNF) which mimics the tumor necrotic action of endotoxin itself. TNF-positive serum is as effective as endotoxin itself in causing necrosis of the sarcoma Meth A and other transplanted tumors. A variety of tests indicate that TNF is not residual endotoxin, but a factor released from host cells, probably macrophages, by endotoxin. Corynebacteria and Zymosan, which like BCG induce hyperplasia of the reticulo-endothelial system, can substitute for BCG in priming mice for release of TNF by endotoxin. TNF is toxic in vitro for two neoplastic cell lines; it is not toxic for mouse embryo cultures. We propose that TNF mediates endotoxin-induced tumor necrosis, and that it may be responsible for the suppression of transformed cells by activated macrophages.


Effect of actinomycin D and estradiol on the response to LH-releasing hormone in neonatally androgenized femal rats. The effect of estradiol, actinomycin D, or both, on pituitary response to LH-RH was studied in 6-month-old female rats which had been injected with testosterone propionate on the 2nd day of life. Estradiol significantly augmented the pituitary response to LH-RH. Actinomycin D did not significantly modify basal serum LH levels and pituitary response to LH-RH but did significantly inhibit the augmenting effect of estradiol on such a response.


The variation of indicator dilution curves with velocity profile. A theoretical analysis has been made of the effect of different velocity profiles on the dispersion of a non-diffusable solute in fluid flow down a straight tube. An experimental investigation of this type of dispersion was made by monitoring the transport of radioactively labelled red blood cells in whole blood at flow rates within the non-Newtonian viscosity range. The dispersion curves obtained are consistent with a progressive flattening of the velocity profile as the flow rate is reduced, though a more rapid clearance than that predicted theoretically occurs in the tail region of the curves. The accepted indicator dilution techniques of estimating the flow rate and vessel volumes are considerably in error. An alternative method of accurately estimating the vessel volume is suggested.


The level of GABA in the brain and locomotor behavior. GABA content was measured in the brains of animals injected with AOAA, DPA or Saline. Significant increases in GABA were found in the motor cortex and cerebellum after treatment with both drugs as compared to saline injected controls. Increased GABA levels were associated with interference with the smooth execution of locomotor acts, especially where balancing and coordination of the hind limbs were necessary.


Trehalose and glycogen levels during the initial stages of growth of Candida albicans. Yeast phase cells of Candida albicans were extracted with cold 0.5 M trichloroacetic acid. The only quantitatively significant anthrone-positive material in these extracts was trehalose which was identified chromatographically and enzymatically. Glycogen in the residue was digested with amyloglucosidase to free glucose which was assayed by a glucose oxidase method. Washed cells from a stationary phase culture of C. albicans rapidly decreased their trehalose content during the early phases of growth on a fresh glucose-containing medium. Concurrent increases in total glycogen concentration were of a greater magnitude than could be accounted for by trehalose mobilization. Thus a specific role for trehalose during initial growth is indicated and is compared with other fungal systems. Trehalase activity was measured in situ and showed only minor changes during the time intervals of these experiments in contrast to other reports on S. cerevisiae. A large fraction of the enzyme in C. albicans behaved as external to the protoplasmic membrane. On one type of growth medium C. albicans exhibited (in addition to trehalose) variable amounts of a sugar alcohol in trichloroacetic acid extracts. This has tentatively been identified as arabitol.


Dimensional analysis and theory of biological similarity. From this review we conclude the following: 1) The body weight of an organism is an adequate reference index for the correlation of morphological and physiological characteristics. In comparative physiology, body weight can be recommended as a unifying frame of reference, particularly if the ponderal scale includes several decades, in order to apply logarithmic scales for the variables involved. (See article). 2) The statistical analysis of the experimental data can be represented conveniently by means of the logarithmic equivalent of Huxley's allometric equation (y = a-Wb), which is the most simple and at the same time the most versatile mathematical expression for intra- or interspecies comparisons. The exponents (b) for the allometric equations can be predicted for all biological variables definable in terms of the MLT system of physics (M = mass, L = length, T = time) or of a four-dimensional system MLTt where t = temperature. 3) By means of dimensional analysis and the theory of biological similarity a range of similarity criteria can be established: a) mechanical or dynamic similarity, b) kinematic or biological similarity; and c) hydrodynamic or transport similarity. Most functions obey the so-called biological (kinematic) similarity, particularly when the concept of operational time is introduced into Lambert-Teissier's original theory. 4) A satisfactory correlation (r = 0.99) for 80 empirical allometric exponents (b) describing morphological and physiological characteristics of living beings was found. These results are discussed in relation to Rosen's optimality principles in biology. 5) Organisms should be considered as mixed regimes. This means that no single similarity criterion can predict the allometric exponent (b) of all functions that dimensionally belong to MLT or MLTt systems, despite the fact that in the great majority of cases kinematic similarity will satisfactorily predict the reduced exponent (b). Nevertheless, in some instances mechanical (dynamic) similarity must be applied, and in other circumstances hydrodynamic (transport) similarity. 6) Cellular or molecular levels are not in the domain of the present theory, since neither cell dimensions nor molecular processes (viz., blood viscosity, diffusion capacity) can be predicted by biological similarity criteria.


Technique for chronic electrode or cannula implantation in decorticate animals. A surgical technique is described which enables chronic recording of electrophysiological activity and chemical or electrical stimulation in the decorticated animal. This procedure circumvents the problems of inadequate electrode/cannula patency and of the short-term debilitating consequences  of cortical ablation. These features make it possible to analyze the relationships between neuroelectrophysiology, behavior and the central mechanisms underlying the restitution of function following brain damage.  The procedure appears to be suitable for a wide range of mature laboratory animals and offers several advantages over the few techniques currently available.


Consummatory behavior in meadow voles (Microtus pennsylvanicus) and mongolian gerbils (Meriones unguiculatus). Consumption of water and adulterated fluids was measured under choice and no choice situation in voles and gerbils. Voles consumed more water throughout, and were clearly less able to tolerate challenges presented by adulteration of available solutions in the no choice situation. Intake of both species under choice conditions was more affected (although differentially) by fluid adulteration that it had been when no choice was available. Male and female differences were noted but typically these differences were in magnitude rather than direction of the consummatory response. The data were taken to illustrate the need for comparative studies and careful specification of stimulus conditions surrounding testing so that general principles of physiological and behavioral regulation can be separated from species  specific differences.


Septal lesions in meadow voles and mongolian gerbils: consummatory and investigatory behavior. In order to separate species specific from general effects of septal lesions on consummatory and investigatory behavior we chose to work with male Mongolian gerbils and meadow voles. These two rodent species are different in both the absolute amount of fluids consumed and reactivity to the palatability of the fluids presented in a single tube. Septal lesions did not elevate water consumption of either species. Septal lesioned gerbils tended to consume more sucrose, and septal voles clearly did drink more sucrose than their control counterparts. But only lesioned voles suppressed intake below control levels when presented with quinine. Exploratory behavior, which was measured only in gerbils, was also altered by septal lesions. Measurement of testosterone, seminal vesicles, and ventral marking glands (gerbils only) indicated that septal lesions alter hormonal systems in at least this species. The importance of comparative investigations in elucidating the general function of brain structures was discussed.


Psychosomatic medicine--past, present, and future. "Psychosomatic" is not part of the average person's vocabulary, and most educated laymen know the meaning of "psychophysiological" and "somatopsychic." Without having words for the process, everyone senses that mental processes can influence bodily functioning and, certainly, that the sick body can produce a sick mind. Many explanations have been offered for the mechanisms underlying these processes--theories ranging from the irrational and religious to the philosophical and scientific. Scientific interest in these processes has been a relatively recent development, originating perhaps a few decades ago. It is time to take inventory, to review and consider the scientific history of psychosomatic medicine and the current status of the field. It is also fascinating to speculate on the developments and ramifications of psychosomatic medicine in the future.


The social and psychological reality of European witchcraft beliefs. In recent years there have been a number of books and articles on witchcraft in general and European witchcraft in particular. The majority of these works contain the implicit or explicit assumption that witchcraft was a cruel, irrational delusion that resulted in the deaths of perhaps hundreds of thousands or innocent victims (Anderson, 1970). While I do not for a moment dispute the horrible cruelties and hideous tortures inflicted on many of the accused, I do feel that it is essential to point out that in terms of the culture and belief structure of Late Medieval and Post Reformation Europe, not only was a belief in witchcraft not irrational and a delusion, but also it did in fact make good sense.


A comparative study of haloperidol and chlorpromazine in terms of clinical effects and therapeutic reversal with benztropine in schizophrenia. Theoretical implications for potency differences among neuroleptics. In a double-blind, cross-over study, the comparative therapeutic effects of 6-week courses of two prototypic neuroleptics--haloperidol and chlorpromazine--and the reversal of those effects with benztropine were investigated in a group of 18 schizophrenics. Periodic measurements were made for 32 dimensions of psychopathology, social participation, span of attention, sleeplessness, pulse rate and neurological side effects. The results showed that haloperidol was generally a more effective drug over the period studied. This was particularly apparent in terms of social and emotional responsiveness, communicativeness and cognitive processes. The only superiority of chlorpromazine seemed to be that patients felt less dysphoric on it than they did on haloperidol. Haloperidol also proved to be more rapid in its action. The data failed to support the clinical validity of the distinction often made between "sedative" and "activating" neuroleptics. Consistent with previous reports, benztropine had the effect of diminishing therapeutic response to both neuroleptics. However, haloperidol again proved less susceptible to this effect. The slowness and lesser therapeutic efficiency of chlorpromazine and its greater susceptibility to benztropine reversal were all considered to be due to its built-in anti-cholinergic properties acting in opposition to its antipsychotic activity. The low potency of chlorpromazine-like drugs was attributed to their inherent anticholinergic characteristics. It was suggested that one of the factors determining potency difference among neuroleptics may be the degree of built-in anticholinergic activity.


A longitudinal therapeutic comparison between two prototypic neuroleptics (haloperidol and chlorpromazine) in matched groups of schizophrenics. Nontherapeutic interactions with trihexyphenidyl. Theoretical implications for potency differences. The treatment process with two prototypic neuroleptics--haloperidol and chlorpromazine--and the nontherapeutic effects of trihexyphenidyl on this process were studied in carefully matched groups of ten schizophrenics each, using a "double-blind", repeated-measure, longitudinal research design. Measurements of various aspects of psychopathology, social participation and clinical indices of arousal were made periodically and objective test of cognition and attention were given. The two treatment groups were highly comparable in epidemiological and clinical terms and differed significantly during the baseline period in only one of the 39 parameters. Longitudinal nonparametric analyses showed that significant therepeutic changes tended to occur more quickly and involved a wider spectrum of schizophrenic phenomena with haloperidol than with chlorpromazine. Parametric analyses also indicated that at the completion of the study, haloperidol-treated patients had significant improvement in many more dimensions than the chlorpromazine-treated patients and that the changes with haloperidol were generally of greater magnitude. At the same time, chlorpromazine treatment seemed to be more susceptible to the antagonistic effects of trihexyphenidyl. No differential patterns of responses were noted for the two neuroleptics to provide any clinical validity to the distinction often made between "sedative" and "activating" neuroleptics. These data were in agreement with those from a previous comparative study which had a very different research design and a somewhat different type of schizophrenic population. The clinical and potency differences between the two neuroleptics were again explained on the basis of the fact that chlorpromazine has much stronger built-in anticholinergic properties, which may be acting in opposition to the antipsychotic activity. It was suggested that the degree of inherent anticholinergic activity may be an important determinant of potency differences among presently known neuroleptics. The possible role of cholinergic mechanisms in schizophrenia was discussed.


The effects of methylphenidate and haloperidol on the heart rate and blood pressure of hyperactive children with special reference to time of action. Twelve hyperactive-aggressive children were given single acute doses of placebo, methylphenidate (0.3 mg/kg) and haloperidol (0.035 mg/kg) on three separate occasions in a double blind crossover design with drug order randomized across subjects. Resting heart rates and systolic and diastolic blood pressures were obtained at nine intervals up to 7 hrs and electrocardiograms at 150 min after ingestion. Methylphenidate caused a small trend toward an increment in heart rate and a significant increment in blood pressure, while the changes with haloperidol were minimal. There were no EKG changes. Changes in heart rate were of a minimal nature as compared with those occurring with digestion which tended to obscure drug effects.


Effects of personal interactions and setting on subjective drug responses in small groups. Subjective responses induced by secobarbital and meprobamate were evaluated in 4 X 4 controlled trials in small groups of healthy young adults over 4 months. Drug effects were mainly influenced by attitudes toward the project, personal interactions, and anticipations of the subjects. The effects of some of these factors were similar to, and at least as powerful as those which are usually considered characteristic of sedatives and stimulants. The same factors, however, did not apparently effect objective responses.


Marihuana effects on TAT form and content. In a double-blind study, 72 normal male subjects were given either placebo or marihuana containing 20 mg. Delta-9-tetrahydrocannabinol. Stories written to cards selected from the Thematic Apperception Test did not differ on hostile or sexual content scales between drug and placebo conditions, but 6 out of 10 scales specifically constructed to detect marihuana effects were successful at differentiating the two conditions. Under marihuana the stories had a timeless, non-narrative quality, with greater discontinuity in thought sequence and more frequent inclusion of contradictory ideas. Novelty of content was somewhat increased by marihuana, while relation to the picture, imagery, repetition, and closure were not significantly affected.


W.H.O. sponsored collaborative studies on nutritional anaemia in India. 1. The effects of supplemental oral iron administration to pregnant women. A W.H.O. sponsored collaborative study of the effects of iron supplementation to pregnant women was carried out in Delhi (northern India) and Vellore (southern India). Supplementation was given under supervision from the 26th to the 36th or 38th week of pregnancy. A control group received only placebo; one group received vitamin B12 and folic acid alone; four groups received vitamin B12, folate and a daily iron supplement ranging from 30 to 240 mg of elemental iron as ferrous fumerate, and one further group received 120 mg of iron without B12 or folate. Groups receiving no iron showed a fall in mean stet concentration. Those receiving iron showed a rise in haemoglobin, the best results being in the groups receiving 120 and 240 mg of iron together with vitamin B12 and folate. Even in these groups however there was still a high prevalence of anaemia and iron deficiency at the end of the trial period. Iron alone did not produce as good results as iron plus vitamin B12 and folate. The supplementation had no detectable effect on the birth weight of the children, nor on the haemoglobin concentration of the infants at three months of age. The daily absorption of iron in the pregnant women, as judged from the increase in haemoglobin mass, was not as satisfactory as expected. Possible reasons for this are discussed. It is concluded that to provide these women with adequate iron a daily oral supplementation of 120 mg of elemental iron or more is needed. This can only be achieved by medicinal means. Before supplementation can be recommended on a public health scale, further information regarding the cost and expected benefits of such measures must be obtained.


[Giant hypertrophic gastritis (Ménétrier's disease)]. A case of giant hypertrophic gastritis is described. The atiology, clinical symptoms, histological changes, therapeutic aspects and the differentiation to the reticulo-blastomas of stomach are discussed.


The radiographic size of renal transplants. The lengths of 55 renal transplants outlined by metallic clips were measured on serial abdominal radiographs up to 3 years after operation. The range of normal transplant size was defined as the mean value +/- 2 standard deviations. Using this definition, the maximum increase in length was 5% at 10 days, 10% at 1 month, and 1% additional for each month in the first year. The normal maximum rate of increase in renal length was 0.5% per day, found only in the immediate postoperative period. Sixty per cent of transplants with severe rejection had either an abnormal length or growth rate or both. In chronic rejection and other complications, size and growth were rarely abnormal. An abnormal decrease in renal length was not seen with any complication.


Lymphomas: initial reactivation.

Lymphomas: initial reactivation. A total of 299 patients with Stage IA-Stage IIB nodal lymphoma treated by irradiation were evaluated for initial reactivation. The abdomen was the major site of reactivation for patients with supradiaphragmatic Hodgkin's disease. The non-Hodgkin's lymphomas reactivated in a variable fashion. Reactivation was prompt for patients with reticulum-cell sarcoma and longer for patients with other lymphomas. Initial extranodal manifestations were present in 24%, 51%, and 63% of patients with Hodgkin's disease, lymphosarcoma, and reticulum-cell sarcoma, respectively.


67Ga citrate in renal allograft rejection. Eight renal allograft recipients were examined on 31 occasions following administration of 99mTc sulfur colloid and 67Ga citrate. Transplant accumulation of each agent was compared and collated with the clinical diagnosis. The procedures matched in 25 instances (81%). Gallium accumulated in the graft producing a false positive result in 2 instances of acute tubular necrosis with anuria, and failed to accumulate within the graft in one case of chronic rejection and in 2 instances of acute rejection. Both radioagents failed to accumulate in 3 patients with acute rejection following heparin therapy. These data indicate that 99mTc sulfur colloid is superior to 67Ga citrate in establishing the diagnosis of transplant rejection and also provides more timely information.


Abnormalities in the regional circulations accompanying congestive heart failure. In congestive heart failure, patients appear to have alimited ability to dilate their resistance vessels in skeletal muscle in response to a metabolic stimulus. This is true whether the metabolic stimulus is ischemia, dynamic, or static exercise. The mechanism for this limited arteriolar capacity is at least twofold; an increased sodium content of the vessels as well as an increased tissue pressure which is seen in edematous states. This can be considered a positive compensatory mechanism in that it helps to maintain systemic arterial pressure during exercise when the cardiac output fails to increase normally. If the resistance vessels were to dilate normally, then in the face of a limited cardiac output, exercise syncope would be expected to occur...


Inward calcium current and activation of contraction in mammalian myocardial fibers. Electrophysiological and tracer studies show an increase in Ca permeability of cardiac cell membranes during excitation. The Ca influx (ICa) during the plateau of the cardiac action potential is a voltage- and time-dependent electrogenic process. The following results were obtained by simultaneous recordings of twitch tension (T) and membrane ionic currents under voltage clamp conditions. 1) The threshold potentials for T and ICa are the same. 2) T decreases when the reversal potential by ICa is approached during depolarization. 3) The voltage and time dependences of recovery of ICa and of T from inactivation are very similar. 4) Catecholamines and methylxanthines which exert a positive inotropic effect increase ICa. 5) Substances which reduce ICa (La3+, Co2+, Mn2+, verapamil, and D 600)  cause a negative inotropic effect. Despite these parallelisms the relation between ICa and T is indirect since during repetitive identical depolarizations ICa decreases while T increases (staircase). This can be explained by a store (sarcoplasmic reticulum?) into which Ca ions flow and from which they are released. Other factors like Na-Ca exchange influence the filling of these stores.


Disturbances of transmembrane ionic fluxes and their role in the pathogenesis of cardiac dysrhythmias. Abnormalities of cardiac rhythm are due either to disorders of impulse propagation (delay or block of conduction, re-entry, circus movement, etc.), or to disturbances of impulse formation (dysfunction of ordinary pacemakers and induction of focal activity). In most instances certain disturbances of transmembrane movements of Na or K ions may be considered to be involved basically in the genesis of dysrhythmic cardiac activity. By means of intracellular recordings as well as of voltage clamp measurements and of analogical computations it is demonstrated that characteristic changes of the membrane potential might result from entirely different ionic mechanisms. Furthermore, an attempt is made to present a detailed analysis of permeability changes underlying normal pacemaker activity in Purkinje fibers as compared with the sinus node or with focal activity induced in myocardial fibers by different influences (aconitine, barium ions, stretch, and strong currents).


The regulatory system of the actin-myosin interaction. According to recent views the active troponin (Tn) complex consists of three components distinguishable by their ability to bind Ca2+ (TnC), to combine with troponyosin (TnT), and to inhibit actomyosin ATPase with ot without Ca2+ (TnI). Addition of Tn to an actin-myosin-tropomyosin system produceas a Ca2+ requirement for ATPase activity. Recent developments bearing on the mechanism both in vitro and in vivo are reviewed in order to correlate biochemical, physiochemical, electron microscopic, and physiological findings. The use of various combination of isolated components considerably helps in pinpointing changes that occur within components (conformational changes) and in the relative positions of interacting moieties. Available data on differences between the skeletal and cardiac system are also discussed.


Bioenergetics in clinical medicine. II. Adjunctive treatment with coenzyme Q in periodontal therapy. Eight patients under routine care for periodontitis received oral treatment with a form of coenzyme Q (7 / CoQ10 and 1 / hexahydrocoenzyme Q4). An unchanged plaque score showed the patients cooperated and were under plaque control. The periodontal score decreased (p less than 0.01) on CoQ treatment. Unexpectedly, the periodontal pocket depth decreased (P less than 0.05) on CoQ treatment since all patients were considered candidates for surgical intervention. Healing was so excellent 5-7 days post-biopsy that the biopsy sites were difficult to locate. The healing was viewed as extraordinarily effective. The mean value of the specific activities of the succinate dehydrogenase-coenzyme Q10 reductase of gingival biopsies increased (P less than 0.05) during treatment which could correlate with the extraordinarily healing. Treatment of periodontitis with coenzyme Q should be considered as adjunctive treatment with current dental practice.


[Porto-systemic encephalo-myelopathy].

[Porto-systemic encephalo-myelopathy]. To avoid the dangerous consequences of gastro-intestinal haemorrhages in portal hypertension, portosystemic anastomosis is often performed. These operations unfortunately cause numerous complications among which isolated encephalopathy and encephalo-myelopathy hold pride of place. In spite of their different clinical, biological, therapeutic and anatomo-pathological behaviour, these two complications are undoubtedly the result of a common pathogenesis of which the details are not at present understood. It is, however, known that these involve complex metabolic disorders connected with the direct passage into the caval blood of products from the gut (ammonia, false neurotransmitters, ...). Experiments with rats have shown development of type II Alzheimer glia in the cerebellum after portocaval anastomosis; so far, spinal lesions have not been reproduced (10-76). In conclusion, it must be recognized that, as Lucien Leger (39) wrote, "by creating a new physiopathology, portal decompression raises as many questions as it solves."


Statistical analysis of the reproducibility of the intravenous glucose tolerance test and the serum insulin response to this test in the middle-aged men. The reproducibility of the intravenous glucose tolerance test (IVGTT) and the serum insulin response to this test have been studied in 48 healthy middle-aged men participating in a health screening program. Two tests were done 4-6 weeks apart. The subjects were ambulatory, and no diet prescriptions other than recommendation of an overnight fast were given. Under these circumstances the correlation between the K values of the two tests was low (r equals 0.55), and the equation for linear regression was y equals 0.56 x + 0.61. The intraindividual variation was 21 per cent, expressed as the coefficient of variation. The early increases in glucose-stimulated serum insulin concentrations in the two tests were significantly correlated (r equals 0.87), and this equation for linear regress was y equals 0.81 x + 10. The coefficient of variation intraindividually for this determination was 20 per cent. We conclude that the K values of the IVGTTs must be judged with caution when obtained in ambulatory practice, especially when the result is used for preventive measures and long-term follow-up of asymptomatic individuals.


Turnover in humans of iodine- and cobalamin-labeled transcobalamin I and of iodine-labeled albumin. Purified transcobalamin I labeled with radioactive iodine and cobalamin was used for turnover studies in 17 humans. Mean fractional catabolic rate was 0.15 d-1 and mean distribution ratio was 0.57. No difference was observed between the turnover in a control group and in patients with vitiligo or pernicious anemia. The plasma curves of the iodine and the cobalamin label were identical, indicating that cobalamin was liberated from transcobalamin I only when this protein was degraded. It was further concluded that transcobalamin I was of minor importance for the transport of cobalamin from the gut to the tissues. The turnover of transcobalamin I was correlated to the turnover of albumin in 9 patients studied with both proteins.


The glucose oxidation in isolated leukocytes from female patients suffering from overweight or anorexia nervosa. Leukocytes from controls and patients with either overweight or anorexia nervosa were incubated with [U-14C]glucose in buffer with or without 50 percent serum. The production of [14C]O2 was measured both under phagocytosing and nonphagocytosing conditions. On the average leukocytes from the two groups of patients produced less [14C]O2 than leukocytes from the controls. Sera from controls stimulated the glucose oxidation in control leukocytes more than sera from patients with anorexia nervosa and less than sera from overweight patients. Neither the insulin nor the glucose content of the sera was of importance for the results. We assume that other serum factors may contribute to the reduced glucose oxidation in leukocytes from patients with anorexia nervosa, whereas this is not the case for patients with overweight.


Renal transplantation in the rat. II. In vitro perfusion of rat kidneys before transplantation. The use of rats in renal preservation studies has many advantages: homogeneity, the availability of inbred strains, low cost, and simplicity of handling. Techniques for renal transplantation in the rat are already established. The present work was performed to standardize the isolated hypothermic perfusion of rat kidneys, using the clearing of blood as a reference point. Reproducibility of isolated perfusion was attained, when the donor rat was anaesthetized with Inactin and pretreated with phenoxybenzamine and heparin. Perfusion was performed with constant flow under controlled temperature conditions, and the perfusion pressure was monitored. Comparative studies of the clearance of blood from the cortex and outer medulla are included in this model. Provided the microsurgical technique is mastered, the rat may be used for studies of renal preservation. The developed technique permits the kidney to be perfused under controlled conditions.


Renal transplantation in the rat. IV. Cortical pressure gradients measured in vivo and during isolated perfusion with reference to renal function after transplantation. Pressure characteristics within the renal cortex of rats were studied with a micropuncture technique in vivo and during isolated hypothermic perfusion. Use of a colloid-free perfusate resulted in an interstitial pressure of 30 cm H2O (normally 1-2 cm H2O), thereby producing a high outflow resistance and perfusion pressure. Interstitial pressures only slightly above normal were found when colloids such as Dextran 40 or albumin were added. The main resistance within the renal vascular bed both in vivo and during isolated perfusion was localized to the afferent and efferent arterioles. The increased pressures were most likely due to extravasation of fluid. With regard to the colloid-containing perfusates, the small but significant increase in interstitial pressure could be explained by increased passage of colloids over the capillary membranes. In conclusion, colloid-free perfusion produced a pronounced but transient elevation of the postoperative serum creatinine level in renal transplant recipients.


The constipating effect of diphenoxylate (Retardinr) in ulcerative colitis. A double-blind controlled trial. A double-blind study of 20 patients with ulcerative colitis during treatment with diphenoxylate 5 mg three times daily and placebo is reported. There were two treatment periods of 14 days each, with cross-over technique and randomized sequence. Test variable for constipating effect was the mean number of defaecations per day. The criterion for inclusion of patients was the presence of 4 or more daily bowel movements; 2 patients did not complete  the investigation. Significant difference (p less than 0.01) in constipating effect between diphenoxylase and placebo was demonstrated both during a period of 12 days and a period of 6 days (Tables II and III). The average number of bowel movements were reduced by 0.7 and 1.3 per day respectively. Side-effects during treatment with diphenoxylate were seen in 53% (Table III) with significant difference against placebo (p less than 0.05). On the basis of the small absolute reduction of defaecation frequency side-effects, it is concluded that diphenoxylate has no place in the routine treatment of ulcerative colitis.


Pentagastrin dose-response in peptic ulcer disease. Stepwise increasing doses of pentagastrin (0.166, 1, and 6 mug/kg-hr) were given to 21 healthy volunteers (HV), 22 patients without peptic ulcer disease (MI), and 22, 18, and 19 patients with duodenal (DU), pyloric (PU), and gastric ulcer (GU) respectively. Maximal response (Vmax) and half maximal dose of pentagastrin (Km) were calculated for both acid and pepsin. The ability to secrete acid and pepsin was in PU and GU slightly higher and lower, respectively, than in the non-ulcer groups. Markedly elevated values were found in DU, and the pepsin secretion was relatively more increased than that  of acid. The values for Km suggested a different behavior of the chief cells and the parietal cells in DU. Whereas the sensitivity of the parietal cells appeared to be abnormally high, that of the chief cells tended to be lower than normal. The discrimination between DU and the non-ulcer groups was better by estimation of pepsin than of acid secretion and tended to improve with increasing stimulation. Superiority was however, found for a submaximal dose of pentagastrin for acid secretion. No superiority in discrimination between groups was found for Vmax. Km was poorly reproducible for acid and pepsin and showed little diagnostic significance. The study suggests that pentagastrin dose-response does not provide more diagnostic information than a single dose (1 mug/kg-hr), which is maximal for pepsin and submaximal for acid.


A double blind trial of carbenoxolone and geranyl farnesyl acetate in gastric ulcer. 57 patients with a chronic gastric ulcer were treated as outpatients in a double blind comparison of carbenozolone 100 mg three times daily with the same dose of geranyl farnesyl acetate (Gefarnate). Healing rates for carbenoxolone were better than for gefarnate, but not significantly so. Nearly half the carbenoxolone-treated patients developed hypokalaemia or oedema, whereas no similar side-effects were found with gefarnate. Gefarnate, while not as effective as carbenoxolone, does appear to promote gastric ulcer healing. Its virtual absence of side-effects makes it a safe and useful drug.


Antibodies against equine ALG during treatment of rejection crisis after renal allotransplantation. The development of antibodies to equine anti-human-lymphocyte globulin (ALG) was followed using passive hemagglutination and gel diffusion, and to Forssman antigen using direct sheep erythrocyte agglutination, in a controlled clinical investigation of ALG treatment of established rejection of renal allografts in a total of 12 patients. With passive hemagglutination a weak antibody to ALG was seen only in one case. There was no increase in titer for sheep erythrocyte antigens during treatment, which agrees well with clinical observations in which no anaphylactic reactions were seen.


Reaction of thymus epithelial cells with antibodies to group A streptococcal polysaccharide. Immunofluorescence studies show that thymus epithelial cells react with antibodies to group A streptococcal polysaccharide.


Bone formation in a free, living bone graft transferred by microvascular anastomoses. A quantitative microscopic study using fluorochrome markers. Bone labeling studies with fluorochrome markers (oxytetracycline and DCAF) were carried out in fifteen dogs in which the posterior portion of the ninth rib was transferred, by means of microvascular anastomoses, to a defect fashioned in the mandible. Seven of the dogs received preoperative radiation of the mandible in order to study the performance of the revascularized graft in radiated tissue. In nine animals the bone grafts were labeled in the subperiosteal, the cortical and the endosteal parts, as opposed to the non-labeling of conventional free bone grafts. Determination of the linear bone formation rate in seven dogs (five non-radiated and two radiated), with the use of a scanning microscope photometer, showed no significant difference between the growth rate in the grafts and that in three other skeletal sites studied.


Surgical treatment of eyelid cancer with special reference to tarsoconjunctival flaps. A follow-up on 193 patients. 203 cancers of the eyelid were operated on in 193 patients. 65% of the tumours occurred in males and the peak incidence was at 75 years. 60% afflicted the lower eyelids and 21% the medial canthi. Basal cell carcinoma were found in 182 specimens and spinocellular carcinoma in only 8. Malignant melanoma, Meibomian gland carcinoma and rhabdomyosarcoma appeared each in one patient, the latter representing the only fatal case in eyelid cancer. All cancers were excised with a free margin around 5 mm. The standard procedures of reconstruction of the eyelid defect were a fullthickness skin graft in cases where conjunctiva and tarsus could be preserved (97 cases), and a tarsoconjunctival flap in full-thickness defects of the lower eyelid (58 cases). Other methods used were wedge excision and direct approximation in very small lesions involving the lid margin, an infratarsal island flap from the lower eyelid for medium-sized to large defects in the upper lid and a forehead or scalp flap after exenteration of the orbit. 18/203 cancers recurred and 12 of these were primarily regarded as radically treated. Seven of the latter were reoperated after more than 3 years and may in fact be new tumours. No metastases were found. The various reconstructive procedures are discussed in detail


Reconstruction of the defective mandible. In a clinical material consisting of 31 cases of mandibular defects, caused by tumour resection or by trauma, reconstruction has been carried out by means of a stabilizing titanium splint and autologous bone and marrow transplantation, the longest period of observation being 9 years. The functional results obtained are assessed with reference to the cause of resection. Different technical procedures are described and the objectives and the planning of reconstruction of the lower jaw are discussed.


Effect of phentolamine and propranolol on the survival of experimental skin flaps. The effect of phentolamine and/or propranolol (alpha- and beta-receptor blocking compounds respectively) on the survival of skin flaps was studied in rats. Phentolamine and/or propranolol were injected intraperitoneally every 12 hours, starting one day before the flap operation, and continued for 5 consecutive days. One week after the operation the survival of the flaps was judged. Systemic treatment with either compound alone or in combination resulted in statistically significant increased survival of the flaps. The results demonstrate that both alpha- and beta-adrenergic receptors influence the survival of skin flaps in rats.


[The effect of fluorides on the adhesion of sealants on etched enamel]. The adhesion of composite filling material was examined on etched and fluoridated enamel in vitro. Etching considerably improves the adhesion. Application of 1% NaF and of 1% acid phosphate fluoride after etching insignificantly decreases it. 1.15% aminofluoride does not change the adhesion.


[Enamel etching for conservative dental treatment]. The enamel etching technique has contributed towards better retention and smaller cavities, and towards less microfissures at the cavity margins. The different etching acids and their concentration are described, and their function illustrated by EM photographs. Phosphoric acid of a concentration between 37 and 50% seems to give the best results. The morphology of the etched surface and the measurements of the degree of adhesion of different composites suggest that the use of bonding agents is not imperative. The practical indications for the etching technique in the dental office are described.


Subpopulations of lymphocytes: a review. Human lymphocytes comprise a heterologous population of cells which can be sub-divided on functional criteria or on the basis of surface membrane markers. Although it is widely accepted that human lymphocytes are broadly divisible into thymus dependent (T) and thymus independent (B) cells, recent evidence suggests that at least in humans this distinction may not be as clear cut as previously thought. An assessment of the relative numbers of T and B cells in human peripheral blood and lymphoid tissues can be made utilizing these marker systems. This has led to a reclassification of human lymphoid diseases according to the T and B cell concept, which might be of value in characterising the immunological status in health and disease, and may allow a more rational approach to the therapy of immune deficiency states and lymphoid malignancies.


Late follow-up in spina bifida cystica.

Late follow-up in spina bifida cystica. The further progress of the survivors in 1969, of a group of 150 babies  born with spina bifida cystica in the period 1960-66 is described. Children with a meningocele continued to do well. Those with a myelomeningocele are divided into 2 groups. There were 4 late deaths and a considerable degree of handicap in the group of 23 in whom the spinal cord was exposed on the surface as a plaque at birth. There were 2 late deaths and a much lower incidence of handicap in the group of 25 in whom only ectopic nervous tissue was found in the sac at birth. The incidence and treatment of hydrocephalus is described. Changes in the attitude to the treatment of a neonate with a myelomeningocele are discussed.


Transmission of Creutzfeldt-Jakob disease from man to the guinea pig. Intracerebral and intraperitoneal inoculation of two guinea pigs with biopsy material from a patient with Creutzfeldt-Jakob disease produced a similar fatal encephalopathy characterized by status spongiosus 422 and 512 days after inoculation. Serial transmission of this disease from guinea pig to guinea pig was achieved in subsequent passages.


Ionophore-mediated calcium entry induces mussel gill ciliary arrest. Lateral cilia of freshwater mussel gills, which normally beat with metachronal rhythm, are arrested pointing frontally by perfusion with 6.25 to 12.5 millimolar calcium and 10(-5) molar A23187, a calcium ionophore. Arrest does not occur in either calcium or ionophore and monovalent cations alone. Activity returns with continued perfusion in potassium chloride or calcium chloride, and more slowly in sodium chloride, after removal of ionophore. These results support the hypothesis that a local rise in internal calcium causes ciliary arrest.


Gout in Maoris.

Gout in Maoris. Historical evidence suggests that the Maori people of New Zealand were virtually untroubled by gout or obesity at a time when these disorders, along with other elements of the gouty diathesis, were rife in the best fed and hardest drinking sections of the Northern European population. By the mid 20th century, however, the apparent decline of the gout in Europe and North America and the breakup of the gouty diathesis in those lands had been more than compensated by their large-scale reappearance in the Maori and in other indigenous inhabitants of the Pacific Basin who, at first sight, appeared to have become one large gouty family. Half the Polynesian population of New Zealand, Rarotonga, Puka Puka, and the Tokelau Islands proved to be hyperuricemic by accepted European and North American standards, the associated gout rate reaching 10.2% in Maori males aged 20 and over. The trends towards hyperuricemia and gout, on the one hand, and towards obesity, diabetes mellitus, hypertension, and associated degenerative vascular disorders, on the other hand, which manifest themselves separately in some Polynesian Pacific Islanders, run together in the Maori and Samoan people, presenting a combined problem of considerable importance to the public health. The appearance of these traits under conditions of plenty in the descendants of hardy and wide-ranging Polynesian voyagers, suggests the emergence of a formerly favorable ancestral polygenic variation through selection for survival under harder conditions. This may now have lost its primitive survival value with a paradoxic shift towards increased prevalence of obesity and the gouty diathesis in more affluent environmental conditions. This may now constitute a genetic load, with recent environmentally determined increase in morbidity and mortality rates from degenerative vascular disorders. There is no satisfactory evidence that overproduction of uric acid differs in mechanism from its European counterparts, although more work remains to be done to determine whether there is any difficulty in renal handling of an increased uric acid load. A high Maori morbidity rate from gout and morbidity and mortality rates from associated components of the gouty diathesis in the face of readily available skilled medical advice and care, indicate the need for greater future attention to help education and health care delivery, at least while conditions of plenty continue. Continuation of previous epidemiologic surveillance may then be required in order to provide a continuing index of the effectiveness of these measures, as well as an opportunity for further research into the interrelationships of these associated disorders.


The role of actin in nonmuscle cell motility. Since actin, myosin, and in certain systems, tropomyosin have now been described in nonmuscle cells, it is not surprising that most of the current models of nonmuscle cell motility are simplified versions of myofibril contraction (see Huxley, 1973; Pollard and Weihing, 1974; Spudich, 1974). These models may very well be valid, but it is unfortunate to limit one's thinking by assuming that one particular specialized cell type, skeletal muscle, must necessarily reflect how the generally occurring phenomenon of cell motility in all cells is achieved. I have described in this chapter two systems in which the generation of motion is carried out in association with actin filaments alone. No myosin appears to be present. In one system motion is effected by the rapid polymerization of actin, in the other by changing the packing of actin filaments. My purpose is not only to suggest that alternative mechanisms are possible and should be looked for in the diverse types of actin associated motility present in nonmuscle cells, but also, and this is the main aim of this chapter, I am suggesting that the basic control mechanisms which control motility in nonmuscle cells may be very different from those in skeletal muscle. I have tried to demonstrate in this regard that the assembly and disassembly of the contractile apparatus in nonmuscle cells is probably the basic control mechanism. Unlike skeletal muscle, in which the contractile apparatus is stable, in nonmuscle cells it is extremely labile forming and breaking down rapidly and repeatedly during development. In fact, much of the motion may be attributed not to myosin-actin interactions, which undoubtedly occur, but to the assembly and disassembly of the apparatus itself. Thus I concentrated on control of the assembly of actin in cells, in particular on what determines the proper spatial and temporal coordinates of this assembly. I described how actin in sperm and red blood cell ghosts can be nonrandomly distributed in cells in the nonfilamentous state which, interestingly enough, is not monomeric actin. Instead much of the actin appears to be bound in the cell. I also discussed how the actin may polymerize relative to membranes, how actin filaments may associate with membranes along their lengths, how the polarity of the actin may be determined, and what might the substance be which holds the actin to the membranes.