Evidence that spo0A mutations are recessive in spo0A-/spo0A+ merodiploid strains of Bacillus subtilis. The spo0A locus contains two types of closely linked mutations that block sporulation at stage 0: spo0A mutations (the most pleiotropic of the stage 0 markers) and spo0C mutations. It was previously thought that spo0A mutations were dominant in merodiploids of Bacillus subtilis, whereas spo0C mutations were recessive. We have shown that spo0A mutations were recessive when spo0A-/spo0A+ merodiploids were made in the genetic backgrounds of strains that were resistant to antibiotic produced by the wild-type strain. Reinvestigation of cultures of spo0A-/spo0A+ merodiploids constructed in the antibiotic-sensitive spo0A strain showed that they contained the spo0A allele at a low frequency, and they produced very few haploid Spo- segregants. These facts indicated that the cultures contained mostly homogenotic (spo0A+/spo0A+) cells. The reason for the poor survival of the spo0A-/spo0A+ merodiploids in the genetic background of the antibiotic-sensitive strain was not clear, but several possible explanations were given. It may have been related to the diploid state of other genes in the same merodiploid cells. The previous indication that spo0A mutations were dominant seems to have been based on the properties of a rare class of Spo- segregants that were probably selected in the presence of antibiotic.


Mutarotation of hydrolysis products by different types of exo-cellulases from Trichoderma viride. Mutarotation of products from p-nitrophenyl beta-D-cellobioside and cellopentaitol by two different types of exo-cellulases from Trichoderma viride was investigated. It was found that an exo-cellulase of glucosidase type produced from the former substrate D-glucose which was mutarotated in a downward direction, while another exo-cellulase of Avicelase type produced from the latter substrate cellobiose which was mutarotated in an upward direction.


Isolation of a membrane protein from R rubrum chromatophores and its abnormal behavior in SDS-polyacrylamide gel electrophoresis due to a high binding capacity for SDS. A membrane protein insoluble in water was isolated by gel chromatography in the presence of 0.1% sodium dodecyl sulfate (SDS) from chromatophores of a photosynthetic bacterium, Rhodospirillum rubrum. This is one of the major membrane proteins of the chromatophore. The protein was found to bind about four grams of SDS per gram, a value which is more than twice the amount generally observed with protein polypeptides derived from water-soluble globular proteins. The electrophoretic behavior of the complex between the membrane protein and SDS is abnormal due to this high capacity for binding SDS. Estimation of the molecular weight of this protein by SDS-polyacrylamide gel electrophoresis was thus impossible. Such an anomaly in SDS binding is unlikely to be restricted to the particular membrane protein described in this paper. The possibility of such a deviation from standard behavior in the interaction with SDS should be taken into consideration in studies of other membrane proteins, since SDS is often used both in analytical and preparative procedures.


Dichroism of bacteriochlorophyll in chromatophores of photosynthetic bacteria. The dichroism was measured in films of air-dried and, consequently, flattened chromatophores of Chromatium vinosum, Rhodopseudomonas sphaeroides and Rhodospirillum rubrum. The values (deltaA/A) of dichroism in C. vinosum were found to be -1.05 at 590 nm and 0.75 in the near infrared region. The values of dichroism in R. sphaeroides were -0.70 at 590 nm and 0.80 at 870 nm. The values of dichroism in R. rubrum were -1.45 at 590 nm and 0.97 at 870 nm.


Immunological purification of sea urchin egg tropomyosin. The antiserum against lantern muscle tropomyosin of the sea urchin was prepared, and the presence of tropomyosin in the sea urchin egg was shown by immunodiffusion test between the antiserum and the egg tropomyosin fraction which was prepared according to the purification method for muscle tropomyosin. The sea urchin egg tropomyosin was isolated from the immuno-precipitate formed between the antiserum and the egg tropomyosin fraction. The subunit molecular weight of the egg tropomyosin was calculated to be 29,000.


Independence of proline chemotaxis and transport in Bacillus subtilis. Values of KI for nine proline analogs as inhibitors of proline chemotaxis and of proline transport were determined. Two of them inhibited transport at substantially lower concentrations than chemotaxis; two at substantially higher concentrations. Moreover, mutants, believed to be in the component that binds proline, were isolated that showed a shift of KM for transport to higher concentrations, one as much as 40-fold. However, chemotaxis was virtually unaffected. Therefore, unlike galactose chemotaxis and transport in Escherichia coli, which share the galactose-binding protein, proline chemotaxis and transport in Bacillus subtilis are independent.


Purification and characterization of NADPH-dependent flavin reductase. An enzyme required for the activation of chorismate synthase in Bacillus subtilis. NADPH-dependent flavin reductase (required for the activation of chorismate synthase) was purified to homogeneity from cell-free extracts of Bacillus subtilis. The enzyme has a molecular weight of 13,000 as determined by sodium dodecyl sulfate-gel electrophoresis, is specific for NADPH, and requires a divalent metal ion and either FMN or FAD for maximal rates of NADPH oxidation. The enzyme is able to reduce 2,6-dichlorophenolindophenol (DCIP) in the presence of NADPH and a divalent metal ion. Both catalytic activities were completely inhibited by EDTA. The Km for FMN is 1.25 X 10(-5) M and for NADPH 7.8 X 10(-5) M with oxygen as the final electron acceptor, and 3.85 X 10(-4) M with DCIP as the final electron acceptor. The enzyme was also isolated in association with chorismate synthase and dehydroquinate synthase. The enzyme associated with the complex has the same catalytic properties as the dissociated enzyme except that it requires both a divalent metal ion and FMN for DCIP reduction. Maximal enzyme activity was observed when the enzyme was preincubated with FMN and the divalent metal ion. The enzyme complex is easily dissociable and the dissociation of the enzyme complex resulted in the failure of NADPH-dependent flavin reductase to adsorb to phosphocellulose.


Purification and properties of chorismate synthase from Bacillus subtilis. Chorismatic synthase was purified to apparent homogeneity from Bacillus subtilis. The enzyme required NADPH-dependent flavin reductase, Mg2+, NADPH, and flavin (FMN or FAD) for activity. The molecular weight of chorismate synthase was 24,000 as determined by sodium dedecyl sulfate (SDS)-gel electrophoresis. The enzyme was also isolated in a complex form associated with NADPH-dependent flavin reductase and another enzyme of the aromatic amino acid pathway, dehydroquinate synthase. On SDS-gel electrophoresis, this form was resolved into three bands with molecular weights of 13,000, 17,000, and 24,000. The enzyme complex was easily dissociated and the dissociation resulted in a change in the chromatographic properties of NADPH-dependent flavin reductase which was no longer retained on phosphocellulose whereas chorismate synthase was still adsorbed. Chorismate synthase activity was linear with time and protein concentration, whereas partially purified preparations showed a significant lag period before the reaction took place. Moreover, crude or partially purified enzyme preparations were completely inactivated by dilution and the activity could be recovered by addition of flavin reductase. A possible role of NADPH-dependent flavin reductase in the activation and regulation of chorismate synthase activity is discussed.


Dehydroquinate synthase in Bacillus subtilis. An enzyme associated with chorismate synthase and flavin reductase. Dehydroquinate synthase, the enzyme which catalyzes the conversion of 3-deoxy-D-arabino-heptulosonic acid 7-phosphate (DAHP) to 5-dehydroquinate, has been purified from Bacillus subtilis in association with chorismate synthase and NADPH-dependent flavin reductase. The enzyme was only active when associated with chorismate synthase, whereas the flavin reductase could be separated from the complex with retention of dehydroquinate synthase activity. The enzyme requires NAD and either Co2+ or Mn2+ for maximal activity. The activity was completely inhibited by EDTA. The Km of the enzyme for DAHP, NAD, and Co2+ were estimated to be 1.3 X 10(-4), 5.5 X 10(-5), and 5.5 X 10(-5) M, respectively. Enzyme activity was completely inhibited by NADH and the inhibition was not reversed by the addition of NAD, NADPH and NADP were not inhibitory. The enzyme was unstable to heat and lost all activity at 55 degrees C. A protein fraction which did not adsorb to phosphocellulose was found to inhibit the enzyme.


Prostaglandin endoperoxide synthetase from bovine vesicular gland microsomes. Inactivation and activation by heme and other metalloporphyrins. Prostaglandin endoperoxide synthetase purified to apparent homogeneity from bovine vesicular gland microsomes contained iron far below the equimolar amount and essentially no heme. However, the enzyme required various metalloporphyrins including hematin or several hemoproteins such as hemoglobin. Preincubation of the enzyme with hematin or hemoglobin resulted in the loss of enzyme activity. The enzyme inactivation was protected by tryptophan or various other aromatic compounds. Furthermore, the simultaneous presence of tryptophan brought about activation of enzyme; namely, the enzyme preincubated with heme and tryptophan showed an almost full activity with a heme concentration in the reaction mixture far below the saturating level. Such inactivation and activation of the enzyme were also observed with manganese protoporphyrin. An identical heme requirement, heme-induced inactivation, and activation of the enzyme were observed in three types of reactions catalyzed by the enzyme: 1) bis-dioxygenation of 8,11,14-eicosatrienoic acid to produce prostaglandin G1, 2) 15-hydroperoxide cleavage of prostaglandin G1 to produce prostaglandin H1, and 3) guaiacol peroxidation. When heme was replaced by manganese protoporphyrin, the enzyme catalyzed only the bis-dioxygenation producing prostaglandin G1 and the activities of the latter two reactions were not detectable.


Regulation of the rate of beta-galactosidase synthese by the Bgs and Bgt loci in the mouse. We have developed an assay for the in vivo rate of beta-galactosidase synthesis in mouse tissues to assess the mechanism by which the Bgs and Bgt loci regulate activity levels of this enzyme. Genetically determined differences in liver and kidney beta-galactosidase content reflect equivalent differences in specific rates of enzyme synthesis. We conclude that Bgs and Bgt regulate beta-galactosidase activity by controlling the rate of synthesis of the beta-galactosidase molecule.


Methionine 500, the site of covalent attachment of an active site-directed reagent of beta-galactosidase. The site of attachment to beta-galactosidase of the active site-directed inhibitor, beta-D-galactopyranosylmethyl p-nitrophenyl triazene, was determined. When the enzyme is completely inactivated, 1 mol of the galactopyranosylmethyl group is bound per mol of monomer with retention of the tetrameric structure. After reaction with the [14C]methyl reagent, labeled peptides were isolated and analyzed. The radioactive label was found to be covalently bound to methionine residue 500.


Metabolism of thromboxane B2 in the monkey. [3H8]Thromboxane B2 was biosynthesized and infused into an unanesthetized monkey. Several urinary metabolites were isolated and their structures elucidated using gas chromatography-mass spectrometry. In addition to the major urinary metabolite, dinor-thromboxane B2, a series of metabolites resulting from dehydrogenetion of the alcohol group at C-11 were identified: 11-dehydro-thromboxane B2, 11-dehydro-15-keto-13,14-dihydro-2,3-dinor-thromboxane B2, and 11-dehydro-15-keto-13,14-dihydro-19-carboxyl-2,3,4,5-tetranor-thromboxane B2. 6-(1,3-dihydroxypropyl)-7-hydroxy-10-oxo-3-pentadecaenoic acid was also identified. Three mono-O-ethylated metabolites were formed from thromboxane B2, which in this study was infused in an ethanolic solution. A small quantity of thromboxane B2 was excreted unchanged into the urine.


Characterization of the purified components of a new homologous series of alpha-mycolic acids from Mycobacterium tuberculosis H37Ra. Homologous series of alpha-mycolic acids from Mycobacterium tuberculosis H37Ra were separated according to size as their p-bromophenacyl ester by bonded C18 reverse-phase, high performance liquid chromatography. Mass spectrometry of the prominent components separated by high performance liquid chromatography gave relatively simple spectra and showed a series of components differing by 28 atomic mass units. A total of different structures were identified. The structures were established as follow: (formula see text) where a = 17, 18, 19; b = 10; c = 15, 17, 19, 21; and d = 21, 23. The C76 and C78 acids contained some C24 alpha-branch acid (d - 21), whereas the C80 and C82 acids contained some a = 19 acids. Several new homologous series were revealed.


Amino acid sequence of beta-galactosidase. V. Isolation and sequences of chymotryptic peptides. The amino acid sequence of 72 chymotryptic peptides isolated from 14C-, 3H-labeled carboxymethyl-beta-galactosidase has been determined. A variety of techniques were used in the isolation procedures including separation by solubility, size, and ion exchange and paper chromatography. These peptides contain approximatley 500 amino acids, range in size from 2 to 26 residues, and give overlaps with tryptic peptides of 16 to 55 residues. Peptides from this digest and those reported earlier from tryptic digests account together for the sequence of about 600 of the 1021 residues in the subunit.


Amino acid sequence of beta-galactosidase. VII. Isolation of the 24 cyanogen bromide peptides. All of the 24 cyanogen bromide peptides of beta-galactosidase have been isolated in pure form. Of these 8 ranged in size from 2 to 5 residues and were purified by paper electrophoresis. The 16 large peptides, from 23 to 119 residues, were chromatographed at pH 5.0 on a carboxymethyl-cellulose column in 0.02 M ammonium acetate buffer containing 8 M urea. A number of peptides were obtained in pure from following Sephadex G-50 or G-75 gel filtration. Others were separated on sulfopropyl-Sephadex or diethyl-(2-hydroxylpropylaminoethyl)-Sephadex. There large peptides were obtained in over 50% yield and several others were obtained in more than 25% yield.


Amino acid sequence of beta-galactosidase. VIII. Sequence of the NH2-terminal segment, CNBr peptides 1 to 9, residues 1 to 377. The amino acids in 9 cyanogen bromide peptides have been placed in sequence starting from the NH2 terminus. The peptides account for residues 1 to 377 of the whole protein and include the largest (CNBr7, 119 residues) and the smallest (CNBr1, 2 residues) of the cyanogen bromide peptides. This region contains only 3 of the 20 lysine residues in the polypeptide chain. A high proportion of charged groups are present (28 of 66 arginine, 28 of 60 glutamic acid, and 24 of 65 aspartic acid residues).


Amino acid sequence of beta-galactosidase. IX. Sequence of the central segment, CNBr peptides 10 to 17, residues 378 to 653. The amino acid sequence in the 8 cyanogen bromide peptides comprising the central segment of beta-galactosidase is presented. This portion of the molecule, about 27% of the protein, contains over 40% of the lysine and tyrosine residues and has a slight excess of basic amino acids.


Amino acid sequence of beta-galactosidase. X. Sequence of the COOH-terminal segment, CNBr peptides 18 to 24, residues 654 to 1021. The sequence of the COOH-terminal third (omega) of beta-galactosidase is presented. The size of the 7 cyanogen bromide peptides of this segment is larger on the average, about 52 amino acid residues as compared to an average size of 42 for cyanogen bromide peptides in the whole molecule. Tyrosine, phenylalanine, and valine are low in this segment whereas alanine and lysine are high. This region has a slight excess of basic groups.


Amino acid sequence of beta-galactosidase. XI. Peptide ordering procedures and the complete sequence. The amino acid sequence of beta-galactosidase has been determined. The monomer contains 1,021 amino acid residues in a single polypeptide chain and has a molecular weight of 116,349. All 80 tryptic peptides as well as all 24 CNBr peptides have been isolated in pure form. Evidence is presented for the ordering of the CNBr peptides. The sequence determination was aided by analysis of cyanogen bromide peptides obtained from a polypeptide fragment produced by a lacZ termination mutant strain.


Immunochemical studies of the inactivation of aspartate transcarbamylase by stationary phase Bacillus subtilis cells. Evidence for selective, energy-dependent degradation. The aspartate transcarbamylase of Bacillus subtilis is stable in exponentially growing cells, but undergoes rapid, energy-dependent inactivation when growth is inhibited by nutrient depletion or addition of antibiotics or other inhibitors of metabolism. This inactivation has been analyzed by a variety of immunochemical techniques, including direct and indirect immunoprecipitation of extracts of cells labeled with 3H-amino-acids, microcomplement fixation, and neutralization of enzymatic activity. The ability of the antibody preparation to react with various denatured, chemically modified, and proteolytically degraded forms of aspartate transcarbamylase was demonstrated. All of the techniques showed that cross-reactive protein disappeared from the cells at the same rate as enzymatic activity, and that little or no immunoprecipitable material of lower than native molecular weight was detectable during inactivation. The disappearance of material cross-reactive with aspartate transcarbamylase occurred prior to the increase in protein degradation that normally occurs in stationary B. subtilis cells and proceeded at a rate at least 20 times greater than general protein degradation. The rate of disappearance was unaffected in mutant strains deficient in intracellular protease activity or in cells treated with inhibitors of protein turnover. Aspartate transcarbamylase was shown to be stable in growing cells. We conclude that the inactivation of aspartate transcarbamylase in vivo involves, or is rapidly followed by, selective, energy-dependent degradation of the protein by a system that appears to involve a previously undescribed protease of B. subtilis.


Characterization of an endoribonuclease, RNase N, from Escherichia coli. The properties of the enzyme ribonuclease N were investigated. By comparing the distribution in the cell of RNase N with the bonafide intracellular beta-galactosidase, and the periplasmic alkaline phosphatase enzymes, we showed that RNase N is an intracellular enzyme. Since previous studies suggested that it is an endoribonuclease, it was compared to RNase III, the only other known intracellular endoribonuclease in Escherichia coli. Using homopolymers and co-polymers we found that, while RNase III could digest double-stranded RNA only, RNase N digested single-stranded and double-stranded RNA with similar efficiency. Furthermore, all RNAs used, natural as well as synthetic, were substrates for the enzyme. Using 5 S rRNA as a substrate it was confirmed that the enzyme is an endonuclease. The final products of the reaction of this enzyme are 5'-mononucleotides. The molecular weight of the enzyme is about 120,000 and it seems to contain two subunits which are similar in size. These properties thus differentiate this enzyme from all other known ribonucleases in E. coli.


Mycolic acids. A reinvestigation.

Mycolic acids. A reinvestigation. Mycolic acids derived from the cell walls of Mycobacterium bovis BCG, Mycobacterium bovis Bovinus I, Mycobacterium smegmatis, and Mycobacterium tuberculosis H37Rv have been fractionated as their p-bromophenacyl esters by a two-step high performance liquid chromatographic procedure: 1) adsorption chromatography on 10-micrometer particle size silica gel, and 2) reverse phase partition chromatography on a 10-micrometer particle size support containing a C18 bonded phase. This procedure has resulted in the isolation of approximately 24 mycolic acids from each bacterium (very likely homologs of various mycolate types) instead of the two to four that have previously been described. The implication of these results on the previously determined structures of these fatty acids is discussed.


Regulation of ribosomal RNA synthesis in Tetrahymena pyriformis. Ribosomal RNA is synthesized at constant rate during most of the cell cycle in heat-shock synchronized populations of Tetrahymena pyriformis. Early in each macronuclear S-period the rate of synthesis increases abruptly, concomitant with replication of the genes coding for ribosomal RNA. The increase is prevented by inhibitors of DNA replication, added prior to the S-period. Similarly, in cultures synchronized by starvation/refeeding, inhibition of DNA replication, at the time when the rDNA is replicated, will prevent the normal increase in rate of RNA synthesis which follows refeeding. We conclude that inhibition of rDNA replication interferes with the synthesis of rRNA, and we suggest that with respect to rRNA synthesis a gene dosis effect is operating in fast-growing Tetrahymena cells.


The ultrastructure of cell division in Euglena gracilis. The ultrastructure of mitosis in Euglena gracilis was investigated. At preprophase the nucleus migrates anteriorly and associates with the basal bodies. Flagella and basal bodies replicate at preprophase. Cells retain motility throughout division. The reservoir and the prophase nucleus elongate perpendicular to the incipient cleavage furrow. One basal body pair surrounded by a ribosome-free zone is found at each of the nuclear poles. The spindle forms within the intact nuclear envelope- Polar fenestrae are absent. At metaphase, the endosome is elongated from pole to pole, and chromosomes are loosely arranged in the equatorial region. Distinct, trilayered kinetochores are present. Spindle elongates as chromosomes migrate to the poles forming a dumb-bell shaped nucleus by telophase. Daughter nuclei are formed by constriction of the nuclear envelope. Cytokinesis is accomplished by furrowing. Cell division in Euglena is compared with that of certain other algae.


Whole-blood lymphocyte stimulation assay for measurement of cell-mediated immune responses in bovine brucellosis. A study was conducted to develop an in vitro whole-blood lymphocyte stimulation assay for measurement of cell-mediated immune response in bovine brucellosis. A soluble antigen (BASA) prepared from killed cells of Brucella abortus 1119-3 was used. Cattle infected with B. abortus field strains, B. abortus 19 calfhood- and adult-vaccinated cattle, and nonexposed cattle were tested. Blood was diluted 10-fold in RPMI-1640 medium (without added serum) and cultured with BASA (at a concentration of 2.2 microgram per culture) at varying times of incubation. Results were assayed for [3H]thymidine incorporation into deoxyribonucleic acid. A 6-day period was found to be optimal for incubating blood cultures to achieve maximum specific lymphocyte stimulation. Serological tests and bacteriological isolation attempts were conducted simultaneously with lymphocyte stimulation tests, and there was a significant correlation between cell-mediated immune response and bacteriological findings. There was a significant correlation between cell-mediated immune response and the level of serum antibodies on a group basis, but there was little correlation between the two systems on individual infected animals. Among vaccinated animals there was little or no correlation between cell-mediated immune and humoral responses. The whole-blood assay was found to be simple, fast, sensitive, and reproducible.


Modified method for serological identification of group B streptococci. Extracts from 10- and 40-ml cultures compared favorably for serological identification of group B streptococci. Typing by gel diffusion proved reliable and efficient. These modifications save time, media, and antisera.


Nitrate reductase activity of Mycobacterium tuberculosis and Mycobacterium bovis in the presence of electron donors. The activities of the nitrate reductase enzyme of Mycobacterium tuberculosis, M. bovis, and of BCG were assayed with and without addition of electron donors. M. tuberculosis always reduced nitrate; M. bovis did so only in the presence of electron donors, and BCG did not show enzymatic activity.


Prophylactic treatment of Rocky Mountain spotted fever. Prophylactic treatment of Rocky Mountain spotted fever with a single dose of oxytetracycline was investigated in guinea pigs. Disease was prevented when treatment was administered shortly before expected onset. Relapses occurred when treatment preceded expected onset by 48 h or more.


Comparison of continuous and discontinuous counterimmunoelectrophoresis with immunodiffusion in identification of candida antibody using HS antigen. We have compared continuous counterimmunoelectrophoresis (CCIE) and discontinuous counterimmunoelectrophoresis (DCIE) with immunodiffusion (ID) in the identification of Candida albicans antibody using HS antigen. Eighteen sera with a titer of 1/1 to 1/32 1/32 by CCIE were titered by CCIE and ID. Fewer were positive by CCIE and ID (13 of 18 and 9 of 18, respectively). Furthermore, the titers were lower when measured by these two methods in comparison to DCIE (4 of 18 and 3 of 18 sera had titers greather than or equal to 1/8 in comparison with 7 of 18 greater than or equal to 1/8 by DCIE). Finally, DCIE was found to be much faster than CCIE and ID. After 20 min of electrophoresis, 17 of 21 sera were positive by DCIE, whereas none were positive by CCIE. Even after 90 min, only 10 of 21 were positive by CCIE, whereas 4 of 21 were positive by ID. The increased sensitivity and speed of DCIE over CCIE and ID suggest that DCIE is preferable to these other methods in the measurement of candida antibody using HS antigen.


Comparison of kinetic and end-point diffusion methods for quantitating human serum immunoglobulins. Proficiency testing results were used to compare the kinetic and end-point versions of the single radial immunodiffusion method for quantitating human serum immunoglobulins. Statistical analysis of the results indicated that the results were not normally distributed but that the log normal distribution gives the best fit of any of the well-known frequency distributions. Consequently, statistical analysis of immunoglobulin results must be log transformed before parametric statistical tests can be appropriately applied. In general, there were no significant differences in level, precision, or interlaboratory comparability for these two methods. However, levels were different for participants using Hyland reagents, and better interlaboratory comparability was achieved by the end-point assay for immunoglobulin M. There were no significant differences in reported levels when compared by manufacturer within the same method.


Improved version of the exoantigen test for identification of Coccidioides immitis and Histoplasma capsulatum cultures. A rapid and simple method for extracting specific cell-free antigens of Coccidioides immitis and Histoplasma capsulatum from agar slant cultures was developed. The extracts were analyzed in immunodiffusion tests for the presence of C. immitis or H. capsulatum specific antigens. These extracts were compared with culture filtrates of brain heart infusion broth subcultures in tests with 32 isolates of C. immitis and H. capsulatum and 13 other fungi which might be morphologically confused with them. The studies showed that the slant extracts were as useful as the culture filtrates and allowed more rapid identification of C. immitis and H. capsulatum. In every case, when an identification was made by conventional morphological methods, the immunological test yielded correlating results. The immunological tests were positive for two isolates that were converted to their yeast forms only after 4 months of study by conventional tests. The new procedure permits the identification of C. immitis and H. capsulatum 2 days after receipt of a pure mycelial-form culture.  The test is recommended for the presumptive identification of C. immitis and H. capsulatum cultures.


Discrepancy between results of the Streptozyme test and those of the antideoxyribonulcease B and antihyaluronidase tests. Comparison of the serum titers obtained with the Streptozyme, the antistreptolysin O, the antideoxyribonuclease B, and the antistreptohyaluronidase tests suggested that the Streptozyme test had failed to detect antibodies against streptococcal deoxyribonuclease B and hyaluronidase. Moreover, sera that were negative in the Streptozyme test could be shown by immunodiffusion to possess significant numbers of precipitins against extracellular factors produced by group A streptococci. Follow-up studies on patients with diagnosed streptococcal infections revealed elevated antideoxyribonuclease and streptohyaluronidase titers and increased numbers of precipitation lines without simultaneous increased titers by the Streptozyme test. There is thus a need for stricter control of possible batch-to-batch variations and more careful standardization of the antigen content of the Streptozyme test.


Aminoglycoside-inactivating enzymes in clinical isolates of Streptococcus faecalis. An explanation for resistance to antibiotic synergism. Clinical isolates of enterococci (Streptococcus faecalis) with high-level resistance to both streptomycin and kanamycin (minimal inhibitory concentration >2,000 mug/ml), and resistant to synergism with penicillin and streptomycin or kanamycin were examined for aminoglycoside-inactivating enzymes. All of the 10 strains studied had streptomycin adenylyltransferase and neomycin phosphotransferase activities; the latter enzyme phosphorylated amikacin as well as its normal substrates, such as kanamycin. Substrate profiles of the neomycin phosphotransferase activity suggested that phosphorylation occurred at the 3'-hydroxyl position, i.e., aminoglycoside 3'-phosphotransferase. A transconjugant strain, which acquired high-level aminoglycoside resistance and resistance to antibiotic synergism after mating with a resistant clinical isolate, also acquired both enzyme activities. Quantitative phosphorylation of amikacin in vitro by a sonicate of the transconjugant strain inactivated the antibiotic, as measured by bioassay, and the phosphorylated drug failed to produce synergism when combined with penicillin against a strain sensitive to penicillin-amikacin synergism.No differences were found in the sensitivity of ribosomes from a sensitive and resistant strain when examined in vitro using polyuridylic acid directed [(14)C]-phenylalanine incorporation in the presence of streptomycin, kanamycin, or amikacin. Therefore, we conclude that aminoglycoside-inactivating enzymes are responsible for the aminoglycoside resistance, and resistance to antibiotic synergism observed in these strains.


Ultrasound findings of renal angiomyolipoma. Angiomyolipoma (AML) IS A BENIGN RENAL NEOPLASM. Preoperative diagnosis is important because of the different surgical approaches to benign and malignant tumors. Angiographic findings of AML and renal carcinoma may be similar and differentiation difficult. B-mode ultraslund examination of AML demonstrates a different image pattern which may be helpfou in differentiation.


Ultrasound scanning for monitoring childhood hydronephrosis. Ultrasound scanning of the kidneys in four children with radiologically proven hydronephrosis was undertaken with a new modification: scanning before and after copious intake of fluid. The anteroposterior diameter of the renal pelvis was used as an index of the hydronephrosis. Alterations in the dilatation were followed over a long period. Good agreement with simultaneous IVP was found. Improvement in the degree of hydronephrosis was apparent by reduction and disappearance of the dilatation of the pelvis during fasting as assessed by ultrasonic scanning, although pronounced dilatation was still observed after copious intake of fluid and at IVP. The results indicate that ultrasound scanning is a valuable supplement in the urological evaluation in children and that it can reduce significantly the number of radiological investigations required.


Determination of renal volume by ultrasound scanning. In patients with renal hypertension, determination of renal volume may be valuable for evaluating possible compensatory hypertrophy for correlating renal size to renal function, and for evaluating the transplanted kidney in relation to rejection and its response to therapy. We performed parallel transverse ultrasonic scans through each kidney and computed volume based on the cross-sectional areas outlined on the scans. In 16 autopsy studies a highly significant correlation between calculated and true values were obtained, rs = 0.847, p less than 0.001. The 5th and 95th percentiles of the median divergence were -21 and 27 ml respectively. In 30 healthy subjects, double determinations showed 5th and 95th percentiles of the median interobserver variation of -7 and 3 ml respectively. There was no demonstrable difference between volumes of right and left kidneys, and no difference in relation to sex. Total renal volume was most accurate when correlated with the body weight, rs = 0.698, p less than 0.001. Normal values of total renal volume per kilogram of body weight were 4.3 to 8.0 ml/kg. In normal subjects, the smallest kidney's volume should not be less than 37% of the total renal volume.


Confusing appearance of a dilated jejunal loop. A case report is presented concerning a patient with a dilated loop of bowel that was misinterpreted on ultrasound examination. Attention is called to this entity as part of the differential diagnosis of an intra-abdominal fluid collection demonstrated by ultrasound.


Ultrasonic demonstration of inferior vena caval involvement with right adrenal gland masses. Ultrasonographic findings in 13 right adrenal gland neoplasms were reviewed. Primary malignancy was the etiology in four patients and metastatic malignancy in nine. Anterior displacement and/or indentation of the posterior aspect of the inferior vena cava was present in nine patients. This sign, although nonspecific, helpful in the localization and diagnosis of right adrenal lesions.


Gallbladder wall thickening: a new sign of gallbladder disease visualized by gray scale cholecystosonography. The use of gray scale equipment in a prospective study of radiologically non-visualizing gallbladders, permitted detection of a new echographic aspect of gallbladder disease. Along with even, discrete bile thickening and the presence of sand-sized calculi, thickening of the gallbladder wall incholecystitis is another ultrasonographic sign of gallbladder disease that can be visualized successfully by gray scale ultrasound.


Prenatal diagnosis of placental tumor by ultrasound. A case of placental chorioangioma is reported. To our knowledge, this is the first case diagnosed prenatally by ultrasonography, The complications of chorioangiomas include hydramnios, antepartum hemorrhage and premature labor, fetal demise, low birth weight and toxemia.


Cholelithiasis and acoustic shadowing.

Cholelithiasis and acoustic shadowing. The importance of the acoustic shadow in the ultrasound diagnosis of cholelithiasis has been stressed in recent reports. In vitro studies of gallstones indicate that the presence and magnitude of the acoustic shadow depend on a number of factors and suggest that a group of small stones may be demonstrable even though no acoustic shadow can be seen.


Persistant corpus luteum cyst of pregnancy: sonographic evaluation of cause of third trimester bleeding. Corpus luteum cysts are frequently observed in pregnant patients evaluated with diagnostic ultrasound. They are usually regarded as incidental findings because of their limited size and usual pattern of regression by ten weeks. The presence and persistence of larger corpus luteum cysts or other extrauterine masses may complicate pregnancy. A case of a large persistent corpus luteum cyst associated with third trimester bleeding is presented.


Carcinoma of the pancreas occuring in a child: a case report with description of gray scale ultrasound findings. This is a report of the gray scale ultrasound findings of pancreatic carcinoma occurring in a child. Although this tumor is rare, the preoperative diagnosis can be made if the physician is aware of this possibility. 99 mTc sulfur colloid radionuclide scan, liver spleen scan, gallium scan, and ultrasound examinations are helpful in distinguishing the site of origin of upper abdominal masses.


Use of pyruvate fermentation compared with tetrazolium reduction in the differentiation of group D streptococci. The reduction of 2, 3, 5 triphenyltetrazolium chloride in the original medium of Barnes (Journal of General Microbiology, 14, 57, 1956), and in a modified medium, was compared with the ability to ferment pyruvate as a means of separating Streptococcus faecalis from all other group D streptococci. The tetrazolium reduction test gave an occasional negative reaction with Str. faecalis strains. In addition, a number of strains gave a weakly positive reaction in the test, as did some strains of Str. faecium and Str. bovis. With some batches of tetrazolium, these weak reactions with Str. faecalis were so frequent that interpretation of the results was difficult. On the other hand, all Str. faecalis strains, and no other group D streptococci, gave a positive pyruvate-fermentation reaction in 48 hours.


Effect of experimentally induced marginal periodontitis and periodontal scaling on the dental pulp. Experimental breakdown of the periodontal attachment apparatus was produced in six young adult monkeys to study the effect on the tissue of the dental pulp by (1) periodontitis, (2) scaling and plaque accumulation on exposed root dentin. Periodontal tissue breakdown was induced by the placement of ligatures around the neck of 92 permanent teeth. Subsequent plaque formation caused marked loss of periodontal tissue support, which after a period of 5--7 months amounted to 30--40% of the root length. One group of teeth received no further treatment. Other teeth were subjected to scaling and root planing. Following treatment, plaque was allowed to accumulate for 2, 10, and 30 days on the freshly planed root dentin surfaces. Histologic examination revealed that in comparison to teeth with normal periodontal conditions, 57% of the teeth exposed to periodontitis exhibited pathologic pulp tissue alterations. Secondary dentin formation and/or inflammatory cell infiltrates were observed within localized areas of the pulp subjacent to root surfaces exposed to periodontal tissue destruction. The changes within the pulp were of "mild" nature and only one tooth displayed signs of total pulp necrosis. Lateral canals communicating with both the pulp cavity and the exposed root surface were never detected. Teeth subjected to scaling and subsequent plaque accumulation in comparison with teeth with periodontitis alone exhibited no obvious aggravation or increased incidence of pathologic pulp reactions. The findings show that in the monkey (1) periodontal destruction limited to the cervical half of the root and (2) plaque accumulation on exposed root dentin does not cause severe alteration in the pulp of the roots involved.


Computed tomography of the brain in patients with headache or temporal lobe epilepsy: findings and cost-effectiveness. Computed tomographic (CT) examinations of the brain were reviewed in 85 patients with a chief complaint of headache and in 22 patients with temporal lobe epilepsy in an attempt to assess the yield and cost-effectiveness in these two clinical settings. Lower bounds for the costs of case finding were calculated using probabilistic estimates. In patients with headache and no objective neurologic findings, no abnormal CT examinations were found. On the other hand, 34% of patients with headache and associated objective neurologic findings had abnormal CT studies. No abnormal CT examinations occurred in patients with temporal lobe epilepsy. The calculated lower bounds for the costs of case finding for these groups were $4,363, $500, and $1,846 per patient, respectively.


Ophthalmologic applications of computed tomography in nonhuman primates. The orbital contents of 28 baboons are defined by computed tomography (CT), which provides a simple, reliable, reproducible, and accurate technique for the experimental investigation of the orbit of the nonhuman primate. This method is applied to the diagnosis of exophthalmos. The suprasellar and other basal subarachnoid cisterns are precisely delineated using intrathecal enhancement prior to CT imaging (metrizamide CT cisternography).


Preliminary evaluation of the histotoxicity and radiopacity of lead-containing elastic impression materials. Severe reactions can be caused when elastic impression materials are trapped beneath soft tissues. To develop a rank order of severity of responses, a series of these materials was injected beneath the oral mucosa of a baboon. Alginate and polysulfide rubbers containing lead peroxide produced severe reactions; hydrocolloid, polysulfide without lead, silicone, and polyether produced more mild reactions. The more toxic materials proved to be more radiopaque than the others. Tests for foreign body toxicity should be made a part of future specifications for these materials, and the use of toxic ingredients should be reduced as much as possible. These materials should be made opaque to X rays and of a different color than those of soft or hard tissues.


Characterization of the reaction sequence involved in phospholipid labeling and deacylation and prostaglandin synthesis and actions. The synthesis and release of prostaglandins (PG) primarily involve a series of enzymatic transformations of the fatty acid arachidonic acid. The source of this free fatty acid is thought to be the cleavage by phospholipase A2 of the acyl bond on the 2-position of phospholipids. The liberated arachidonic acid is primarily converted by cyclo-oxygenase (PG-synthetase) into the PG-endoperoxides (PGG2 and PGH2). These compounds represent a critical branch point in PG metabolism since they serve as substrates for the enzymatic synthesis of PGE2, PGD2, thromboxane A2, PGI2 (prostacyclin), as well as some likely products yet to be identified. The ultimate fate of arachidonate in a given tissue is determined by the repertoire of enzymes present. The prostaglandins have a very broad sprectum of identified biological activities and the newly discovered arachidonate metabolites (endoperoxides, thromboxane A2 and PGI2) are labile and extremely potent.


Marrying regulatory and competitive approaches to health care cost containment. The aim of this paper is to demonstrate that the problems which arise from simultaneously developing regulatory and competitive approaches to health care cost containment can be solved, if recognized, and that those problems deserve more systematic investigation than they have so far received. It is suggested that public regulation which accommodates the special characteristics of health maintenance organizations (HMOs) allows for effective competition. Although there are formidable difficulties in formulating the appropriate regulatory strategies, there are some encouraging signs, such as the renewed commitment of the Carter Administration and Congress to the development of HMOs and the active interest of the Federal Trade Commission in health care, that future policy may minimize conflict between regulation and competition.


Isolation and analysis of H-2 and Ia alloantigens from wild mouse strains. Immunoprecipitation and SDS-PAGE analysis were used to examine H-2 and Ia antigens from mouse strains with wild-derived MHC haplotypes. Antisera raised against the wild-derived strains contained anti-H-2 and anti-Ia antibodies which precipitated antigen molecules readily distinguishable by a single assay procedure. The antibodies to wild strains were cross-reactive with standard laboratory haplotypes. Evidence supporting the similar genetic organization of wild and standard haplotypes was found, including isolation of separate H-2K and H-2D molecules from a wild-derived strain, and isolation of two separate Ia molecules from a wild-derived strain.


Light chain diversity of murine anti-streptococcal antibodies: IgCH-linked effects on L chain expression. L chains derived from anti-group A streptococcal carbohydrate antibodies raised in A/J, BALB/cJ, C57BL/6J, CB-20, BAB-14, and CAL-20 mice were examined by isoelectric focusing. Multiple strain-associated differences in the degree and frequency of expression of particular L chain spectrotypes were observed. Analysis of L chain-focusing patterns in allotype-congenic mice revealed that IgCH-linked genes can have profound effects on the L chain phenotypes expressed by strains with identical L chain genotypes. Lastly, the overall spectrotypic diversity of L chains from anti-GAC antibodies appears to be less extensive than the diversity of the antibodies from which these L chains derive, documented by similar techniques. These results are interpreted in light of the significance of combinatorial diversity in generating antibody heterogeneity.


Elimination of inter-species reactive anti-IgG antibodies by affinity chromatography. Inter-species reactive anti-IgG antibodies are disturbing in blocking and interference experiments with the sensitive mixed hemadsorption (MH) technique. By affinity chromatography this disturbance could be efficiently eliminated. Anti-IgG sera were passed through columns containing heterologous IgG. The absorption technique is rapid and simple. After one absorption the inter-species reactivity in anti-IgG sera had been decreased thousand-fold, while any loss of species-specific reactivity could not be detected. Such absorbed anti-IgG sera reacted only with homologous antibodies. The effect is exemplified with e.g. blocking of human HLA antibodies by anti-beta2-microglobulin from rabbits.


Preparation of antibody fragments: conditions for proteolysis compared by SDS slab-gel electrophoresis and quantitation of antibody yield. Monovalent antibody fragments are now recognized to have major advantages over intact immunoglobulin G in immunohistochemistry, but the methods commonly used for their preparation do not necessarily give a maximal yield of active antibody fragments and take little account of species differences. Sodium dodecyl sulphate (SDS) slab-gel electrophoresis demonstrated large differences in the susceptibility to digestion of antibodies of different species and was found to be valuable in the selection of optimal digestion conditions. Promising results were obtained with sheep antibodies by use of low concentrations of trypsin, and an immunoadsorbent technique showed a high yield of antibody fragments.


Separation and characterization of anti-benzylpenicilloyl (BPO) antibodies. II. Immunological properties of different IgG fractions. Affinity chromatography and subsequent ion-exchange chromatography of pooled anti-benzylpenicilloyl (anti-BPO) hyperimmune sera separated 5 different anti-BPO IgG fractions as described in the preceding paper. These fractions were tested for activities in passive hemagglutination (PHA), passive immune hemolysis (PIH), antibody-dependent cellular cytotoxicity (ADCC) and influence on IgM-induced hemolysis. It was found that anti-BPO IgG fractions with low avidity (dissociation constant K = 4.4--6.7 X 10(-8) moles/l) were poorly active in PHA and ADCC and had no blocking activity in IgM hemolysis. Among the highly avid antibodies (K = 0.7--3.4 X 10(-8) mole/l) no correlations were found between avidity and activities in the immunological tests. The results presented demonstrate that ion-exchange chromatography allows the separation of blocking and lytic antibodies as shown by their influence on IgM-induced complement-dependent lysis of lightly hapten-coated sheep erythrocytes.


A simple routine method for detecting hidden rheumatoid factors. A new and simple routine method is described for detecting hidden rheumatoid factors in human serum. EDTA glycine and NaCl were used to liberate hidden rheumatoid factors and to inactivate complement before rheumatoid-factor activity was determined in a glycine--NaCl solution. Forty-nine out of 97 sera from individuals with seronegative rheumatoid arthritis gave positive reactions by this method. Rheumatoid sera with low titres by standard tests gave higher titres with the new method. The new method detects both IgM and IgG rheumatoid factors and is simple and suitable for use in routine medical laboratories. Used in parallel with the classical tests, it facilitates detection of hidden rheumatoid factors.


Contribution of omega-oxidation to fatty acid oxidation by liver of rat and monkey. Contributions of omega-oxidation to overall fatty acid oxidation in slices from livers of ketotic alloxan diabetic rats and of fasted monkeys are estimated. Estimates are made from a comparison of the distribution of 14C in glucose formed by the slices from omega-14C-labeled compared to 2-14C-labeled fatty acids of even numbers of carbon atoms and from [1-14C]acetate compared to [2-14C]acetate. These estimates are based on the fact that 1) the dicarboxylic acid formed via omega-oxidation of a omega-14C-labeled fatty acid will yield [1-14C]acetate and [1-14C]succinate on subsequent beta-oxidation, if beta-oxidation is assumed to proceed to completion; 2) only [2-14C]acetate will be formed if the fatty acid is metabolized solely via beta-oxidation; and 3) 14C from [1-14C]acetate and [1-14C]succinate is incorporated into carbons 3 and 4 of glucose and 14C from [2-14C]acetate is incorporated into all six carbons of glucose. From the distributions found, the contribution of omega-oxidation to the initial oxidation of palmitate by liver slices is estimated to between 8% and 11%, and the oxidation of laurate between 17% and 21%. Distributions of 14C in glucose formed from 14C-labeled palmitate infused into fasted and diabetic rats do not permit quantitative estimation of the contribution of omega-oxidation to fatty acid oxidation in vivo. However, the distributions found also indicate that, of the fatty acid metabolized by the whole animal in the environment of glucose formation, at most, only a minor portion is initially oxidized via omega-oxidation. As such, omega-oxidation cannot contribute more than a small extent to the formation of glucose.


Isolation and characterization of flightless mutants in Drosophila melanogaster. Since animal behaviour is executed through neuronal circuits including sensory receptors and muscle, genes vital for their development and differentiation must be found among mutants having behavioural anomaly. After mutagenesis with ethyl methanesulphonate (EMS), we screened for X-linked flightless mutants of Drosophila melanogaster by using column-type flight tester. Approximately 10(4) individuals were screened and 21 mutant genes were isolated. Chromosomal mapping and complementation experiments revealed that they belong to 15 cistrons randomly located on X chromosome, three cistrons having more than two alleles. Two of the isolated mutants (fltO2 and fltH, which are recessive both behaviourally and morphologically) were analysed with the mosaic fate mapping technique, and both were found to have their primary foci in mesodermal region of blastoderm, suggesting that the genes exert their primary effect in indirect flight muscle. Electronmicroscopic studies on the muscles from four alleles of the fltO2 cistron revealed an abnormality in myofibrillar arrangement. A possible deficit within Z-band components is discussed in relation to wings-up B mutants. The indirect flight muscle of flltH was also examined, and it was found that sarcomere length and diameter of myofibrils were abnormal. It was postulated that a possible factor which controls size of myofibrils is defective in this mutant. These examples indicate the advantage of combining ultrastructural examination with genetic mosaic mapping technique.


Cell culture of individual Drosophila embryos. I. Development of wild-type cultures. A new procedure is described for the preparation of in vitro cell cultures from individual early gastrulae of Drosophila melanogaster. In these cultures several identifiable cell types differentiate within 24 h (nerve, muscle, fat-body, haemocyte and chitin-secreting); their initial appearance and continuing development over a period of weeks is described. It is proposed that this technique may be used to analyse abnormalities of cellular development in embryonic lethal mutants. Culture in vitro of cells from lethal embryos is seen to have two broad roles: (1) to test the developmental capacity of individual cell types in a situation where they are relatively free from possible deleterious interactions with other cell types and are liberated from the system of the dying embryo, and (2) through the preparation of mixed cultures from normal and mutant embryos, to determine the influence of the presence of wild-type cells on observed abnormalities of a particular cell type.


Cell culture of individual Drosophila embryos. II. Culture of X-linked embryonic lethals. Results are reported from the culturing in vitro of cells from individual early gastrulae of the following four groups of X-linked embryonic lethal mutants of Drosophila melanogaster. (1) Notch lethals, Five Notch mutants were studied which have been reported to give similar abnormalities in whole embryos: the nervous system displays a three-fold hypertrophy as part of a shift in the pattern of differentiation within ectodermal derivatives, and mesodermal derivatives do not differentiate. An hypertrophy of nerve was found in cell cultures prepared from embryos of all five mutants. In addition, four of the five alleles consistently gave abnormalities of muscle differentiation: when compared to controls, Notch cultures had a reduced frequency of myotubes, and displayed unusual clusters of myocytes which had either failed to fuse or had fused incompletely. Results from mixed cultures prepared from two embryos were consistent with the autonomous expression of nerve and muscle abnormalities by Notch-8 cells in the presence of wild-type cells. It is argued that the Notch locus has a direct role in the differentiation of both nerve and muscle. (2) white deficiencies. Cells carrying either of two deficiencies gave a clear-cut pattern of abnormalities: initial cellular differentiations were normal, but nerve, muscle and fat-body cells progressively deteriorated during the culture period. Mixed cultures showed that wild-type cells could not 'rescue' mutant muscle and fat-body cells; however, the status of the autonomy of mutant nerve abnormalities in these cultures was unclear. Both white deficiencies remove cytological band 3C1, and this permits a comparison of results with those from cultures of cells from Notch-8 embryos (also deficient for 3C1). Abnormalities displayed in cultures of the two types of mutant show no overlap. Therefore no consistent cellular abnormality can be attributed to absence of band 3C1. (3) lethal(1)myospheroid. In contrast to earlier observations on in vitro cell cultures (Donady & Seecof, 1972) muscle was seen to differentiate, though its morphology was extremely abnormal. Observations indicated that all cell types within the cultures had poor properties of adhesion to a glass substrate. It is argued that the observed abnormalities are not consistent with a mutant lesion which is restricted to the basement membrane (contra Wright, 1960), and that all cell types carry a basic defect which may reside in the cell membrane. (4) shibirets alleles. Cultures of two temperature-sensitive lethal shibire alleles (shits1), shits3) were normal at the permissive temperature of 22 degrees C. At the restrictive temperature (29 degrees C) early cell differentiation was normal but subsequent development was blocked. This blockage could be partially reversed by shifting cultures to the permissive temperature after as much as 10 days exposure to the high temperature. It is suggested that shits cells are mutant in a process which is basic to several cell types.


Scanning electron microscopic observations of the development of the somites and their innervation in anuran larvae. The development of the paraxial mesoderm in tadpoles of Xenopus, Bufo and Rana was observed with a scanning electron microscope. In addition to examination of the differentiation of the surface and the interior of the somites, some attention was also paid to the transformation of the material of the neural crests and to the innervation of the developing myotome.


Control of immunoglobulin secretion in the murine plasmacytoma line MOPC 315. Cells of the 315LV-1 (derived from NP1) variant line of MOPC 315 contain approximately 1% the normal intracellular level of the heavy (alpha) chain of IgA and no detectable light (lambda2) chain. The synthesis rate of alpha-chain in the variant, however, is similar to that in cells of the parent line. Moreover the relative amount of translatable alpha-chain mRNA that can be extracted from 315LV-1 cells is about the same as for parental cells. No light-chain synthesis can be detected either in vivo or in vitro in a wheat germ cell-free system. The 315LV-1 heavy chain synthesized in vivo or in vitro has slightly greater electrophoretic mobility than normal H chain and turns over rapidly intracellularly. The variant fails to secrete any of its heavy chain, despite the fact that its H chain mRNA is bound to membranes, as one would expect for a secretory protein message. Fusion of 315LV-1 cells with cells of a kappa-producing MPC 11 variant line leads to stabilization of the intracellular H chain and also to full recovery of secretion of the H chain as an H2L2 molecule.


Acquisition of murine major histocompatibility complex gene products by schistosomula of Schistosoma mansoni. Schistosoma mansoni schistosomula recovered from the lungs of inbred mice were shown to possess serologically detectable alloantigens on their tegumental surfaces. Using appropriate antisera and infected congenic and recombinant mice as worm donors, gene products of the K and I subregions of the major histocompatibility complex were demonstrated among these alloantigens acquired by the parasites. In contrast, other cell surface alloantigens, such as Thy 1, Ly 1, and H-Y and the serum proteins albumin, C3 and Ig, could not be detected on the surface of lung schistosomula by means of comparable techniques. In another series of experiments, schistosomula recovered from the lungs of mice and reinjected into allogeneic recipients were shown to exchange their alloantigens during an 87-h period of examination. Similarly, lung schistosomula cocultured with allogeneic lymphocytes were shown to acquire major histocompatibility complex (MHC) coded antigens from the cells. It is possible that as acquired host molecules, MHC gene products may disguise the surface of schistosome parasites thereby rendering them insusceptible to immune attack.


[Elaboration of a method for the quantitative determination of proetins by laser nephelometry in the clinical routine laboratory (author's transl)]. The reaction conditions for the quantitative determination of IgC, IgA and IgM in the laser nephelometer were investigated. An easy working scheme was devised. The method was compared with radial immunodiffusion, and it was statistically evaluated.


Characterization of the effects of Mg2+ on Ca2+- and Sr2+-activated tension generation of skinned rat cardiac fibers. Submaximum and maximum forces of the cardiac muscle contractile apparatus, activated by Ca2+ or Sr2+, were determined as a function of Mg2+ concentration. Apical left ventricular tissue from Sprague-Dawley rats was broken by homogenization into small bundles of fibers with disrupted sarcolemmas (skinned). Tension generation was activated by and graded according to the concentration of Ca2+ or Sr2+ in solutions bathing the skinned fibers and measured with a photodiode force transducer. Steady-state tensions for various levels of activation at each of four concentrations of Mg2+ (5 x 10(-5), 1 x 10(-3), 5 x 10(-3), and 10 x 10(-3) M) in the bathing solutions were analyzed. Other bathing solution constituents and parameters mimicked significant normal intracellular conditions while providing adequate buffering of [H+], [Ca2+], and [MgATP2-] (magnesium adenosine triphosphate). To assess changes in sensitivity of the mechanical system to activation by Ca2+ (or Sr2+), each submaximum tension was expressed as a percentage of the given fiber bundle's maximum force generated at saturating [Ca2+] (or [Sr2+]) at the same [Mg2+]. When plotted as saturation curves these data demonstrate that increasing [Mg2+] depresses Ca2+ sensitivity of the force-generating mechanism. The Ca2+ and Sr2+ sensitivity of the cardiac force-generating apparatus is similar at every [Mg2+], indicating that the magnitude of Mg2+ effect is similar for both types of activation. However, absolute maximum tensions at saturating activating cation concentration increased as [Mg2+] increased; the effect of Mg2+ on maximum force was proportionately the same for Ca2+ and Sr2+ activation. But because saturating [Ca2+] always resulted in a lower maximum force than saturating [Sr2+], this site of Ca2+-Mg2+ interaction appears distinct from the one influencing Ca2+ sensitivity.


Multiple ophthalmic anomalies and digital hypoplasia. Multiple congenital eye and hand anomalies occurred in a young female born to normal but consanguineous parents. Both eyes were microphthalmic with severe corneal, iris lens pathology. Ultrasonography revealed multiple echos from the vitreous. The ocular findings are suggestive of retinal dysplasia. A skeletal dysplasia, presenting as distal phalangeal hypoplasia, was found in both hands. There was no history of intrauterine exposure to drugs. This appears to be a unique association of congenital malformations, without other systemic involvement. Diagnostic and genetic implications are discussed.


Experimental infection of chimpanzees with the Norwalk agent of epidemic viral gastroenteritis. A fecal filtrate of human origin containing the Norwalk agent of epidemic viral gastroenteritis was administered by stomach tube to chimpanzees in an attempt to induce diarrheal disease. Significant postchallenge serum antibody rises against Norwalk viral antigens were demonstrated in all animals using the techniques of immune electron microscopy and radioimmunoassay. In addition, viral antigens were detected in feces from five of nine animals using radioimmunoassay. Clinical illness characterized by diarrhea and/or vomiting did not occur. Infection was transmitted subsequently by feeding four additional chimpanzees a fecal filtrate prepared from one of the previously infected animals. Development of an antibody response in four animals and detection of viral antigen in two animals that received this passage filtrate indicated that viral replication had occurred in the absence of clinical illness. The availability of the chimpanzee as an experimental animal host susceptible to infection with the Norwalk agent should facilitate the study of epidemic viral gastroenteritis.


Solid-phase microtiter radioimmunoassay for detection of the Norwalk strain of acute nonbacterial, epidemic gastroenteritis virus and its antibodies. The development of microtiter solid-phase radioimmunoassays for the detection of Norwalk antigen and its antibody is described. The tests are simple to perform and are sensitive and specific. The test for antigen can be used on crude stool filtrates and suspensions. Both tests are at least as sensitive as immune electron microscopy and more sensitive than immune adherence assay.


Menkes' disease: neurophysiological aspects. The evolution of neurophysiological features including the electroencephalogram (EEG), electroretinogram (ERG), and visual evoked potentials (VEP) is reported in eight cases of Menkes' "kinky hair" disease. All EEGs were severely abnormal, with some characteristic features seen from 3-5 months of age, after the onset of clinical symptomatology. From the age of 5 months, the EEGs resembled hypsarrhythmic patterns. The ERG was not affected in any patient, but the VEP was either of low amplitude or completely absent in all but one of the six patients tested. All eight patients received copper injections without substantial effect on either the clinical course of the disease or the EEG features.


Lymphocyte responsiveness to Pseudomonas aeruginosa in cystic fibrosis: Relationship to status of pulmonary disease in sibling pairs. Lymphocyte proliferative responses to Pseudomonas aeruginosa and Staphylococcus aureus were evaluated in six sibling pairs with cystic fibrosis. In each pair, one sibling had advanced clinical disease, whereas the other sibling was in good clinical condition. Three in this latter group had no clinically apparent Pseudomonas bronchitis. In all cases, the average responses to Pseudomonas isolated from each sibling pair were lower in the sibling with advanced clinical disease. This difference was not observed in the responses to Staphylococcus. Normal plasma or plasma from patients with CF in good clinical condition does not restore the responses in patients with advanced clinical disease. However, plasma from patients with low or no responses to Pseudomonas inhibits the responses of responding siblings. A progressive specific lymphocyte unresponsiveness to Pseudomonas may play an important role in the increasing destructiveness of chronic pulmonary Pseudomonas infection in cystic fibrosis.


The strength of the reflex response to sinusoidal stretch of monkey jaw closing muscles during voluntary contraction. 1. Rhesus monkeys were trained to exert steady biting forces of 3--60 N for 1--2 sec. This behaviour was well maintained while sinusoidal or step opening and closing movements were imposed on the jaw. 2. The amplitude of the force modulation during sinusoidal stretching was divided by the amplitude of movement to obtain the magnitude of stiffness. This estimate was made at frequencies from 2 to 50 Hz at amplitudes of 100 and 500 micrometer (half the peak-to-peak movement at the incisors). 3. Peak magnitudes of stiffness were seen with frequencies of 8--15 Hz when the amplitude of movement was small; there was a great deal of variation between individual animals. This variation was most striking with mean forces of 25--35 N. The stiffness was greatest in animals that showed considerable spontaneous tremor, and the highest levels of stiffness were often recorded with frequencies near which tremor amplitude was large. A marked phase lag in the force response was often seen during small amplitude stretching at 8--30 Hz. 4. Estimates of stiffness for larger amplitude (500 micrometer) stretching showed less variation; the magnitude of stiffness showed maximum values below 10 Hz and a minimum at 15--30 Hz. Force always showed a phase lead on position although this lead became small in the frequency range where with smaller movement there had been phase lags. The magnitude of stiffness increased with increasing mean force. 5. Bilateral electrolytic lesions were made in the brain stems of three animals; they reduced by over 95% the expected number of cells in the mesencephalic nucleus of the fifth cranial nerve on either side. These lesions interrupted the afferent pathway for the stretch reflex and so abolished excitatory electromyogram (e.m.g.) responses to step stretches of the jaw closing muscles. 6. Such reflex responses as persisted after the lesions were small and inhibitory. E.m.g. silences followed both step stretch and release; the response to release was a 'load compensation' that could not be attributed to spindle afferents. 7. After the lesions the responses to movements of 100 micrometer showed neither negative values for the phase nor marked peaks in the stiffness magnitude at low frequencies; these features therefore take origin in the action of the stretch reflex. The stiffness that was measured after the lesions may be attributed to the non-reflex components resisting stretch, particularly to the properties of the contracting muscles. Thus, the phase of the force response was markedly advanced at all frequencies and the stiffness seen for 100 micrometer was similar to that for 500 micrometer. Stiffness increased with increasing mean force, as before surgery. 8. Vector subtraction of the stiffness seen at each frequency after interrupting the stretch reflex from that seen before doing so gave a quantitative estimate of the strength of the stretch reflex. The reflex activity calculated in this way showed attenuation and progressive phase lag as the frequency increased above 10 Hz...


Synthesis and evaluation of bis-dipeptide and bis-tripeptide analogues of actinomycin D. Six-bis-dipeptide analogues of actinomycin D, all containing two threonyl-D-valine side chains, were prepared. Also two bis-tripeptide analogues containing an additional proline or oxoproline residue were synthesized. None of the compounds bound to DNA in a manner similar to actinomycin D. This lack of strong intercalative binding emphasizes the importance of the pentapeptidolactone side chains in the binding of actinomycin D to DNA and also highlights the deficiences inherent in using only small nucleotide sequences in investigating drug-DNA binding. None of the analogues tested showed any antitumor activity, although actinocylbis(threonyl-D-valine methyl ester) did show 10% of the antibacterial activity of actinomycin D vs. Bacillus subtilis.


Folate antagonists. 12. Antimalarial and antibacterial effects of 2,4-diamino-6-[(aralkyl and alicyclid)thio-, sulfinyl-, and sulfonyl]quinazolines. A series of 2,4-diamino-6-[(aralkyl and alicyclic)thio-, sulfinyl-, and sulfonyl]quinazolines was prepared via condensation of 5-chloro-2-nitrobenzonitrile or 5,6-dichloro-2-nitrobenzonitrile with the appropriate aralkyl or alicyclic thiopseudourea, reduction of the resulting 2-nitro-5-[(aralkyl or alicyclic)thio]benzonitrile with stannous chloride to the amine, and cyclization with chloroformamidine hydrochloride. Oxidation was effected with hydrogen peroxide or the bromine complex of 1,4-diazabicyclo[2.2.2]octane. These analogues when examined for suppressive activity against drug-sensitive lines of Plasmodium berghei in mice were not as active as 2,4-diamino-6-[3,4-dichlorobenzyl)amino]quinazoline (Ia).


trans-2,3b,4,5,7,8b,9,10-Octahydronaphtho[1,2-c:5,6-c']dipyrazole, a new orally active antiallergic compound. The synthesis and antiallergic activity of a new heterocyclid steroidal molecule are described. Compound 1 was shown to inhibit the rat passive cutaneous anaphylaxis and its activity in this system was compared to that of disodium cromoglycate. It is orally active at a dose of 35 mg/kg (ED50) and its activity persists for up to 6 h for larger doses.


Acute myelogenous leukaemia in Hurler's syndrome. The occurrence of the Hurler syndrome and acute myelogenous leukaemia in a 2 1/2-year-old girl is described. This represents the first published report of the concurrence of these two diseases.


Effects of 3'deoxyadenosine and actinomycin D on RNA synthesis in toad bladder: analysis of response to aldosterone. Previous studies have shown that aldosterone increases transepithelial active Na+ transport after a latent period of about 60 min and incorporation of 3H-uridine into polyadenylated RNA (poly(A)(+)RNA) (putatively poly(A)(+)mRNA) as early as 30 min after aldosterone addition. To assess the physiological importance of this pathway, the effects of 3'deoxyadenosine and actinomycin D were compared in studies on the urinary bladder of the toad Bufo marinus. 3'deoxyadenosine (30 microgram/ml) only partially, though significantly, inhibited the aldosterone-dependent increase in Na+ transport measured as short-circuit current (scc). The incorporation of 3H-uridine into poly(A) (+)RNA was inhibited by 70 to 80%. In contrast, Actinomycin D (2 microgram/ml) totally inhibited the aldosterone-dependent increase in scc, and the incorporation of 3H-uridine into poly(A)(+)RNA by 68 to 75%. 3'deoxyadenosine or actinomycin D alone had no significant effects on baseline scc, while inhibiting poly(A)(+)RNA to the same extent. The differential effects of deoxyadenosine and actinomycin on aldosterone-dpendent Na+ transport may be related to their different sites of action on RNA synthesis: both drugs inhibited, to a similar extent, cytoplasmic poly(A)(+)mRNA: however, 3'deoxyadenosine, in contrast to Actinomycin D, failed to inhibit poly(A)(-)RNA, sedimenting between 4S and 18S (putatively poly(A)(-)mRNA). We conclude that the mineralocorticoid action of aldosterone during the first three hours depends on the synthesis of both poly(A)(+)mRNA and poly(A)(-)mRNA.


Ionic and metabolic requirements for the hydroosmotic response to antidiuretic hormone in toad urinary bladder. A study has been conducted to determine the ionic and metabolic requirements for full expression of the hydroosmotic response to antidiuretic hormone in the toad urinary bladder. By appropriate manipulation of incubation conditions it can be shown that there is a pool of serosal sodium necessary for a full hormone response. This serosal sodium pool is not related to the transepithelial sodium transport pool A full hydroosmotic response also requires serosal potassium; however, no specific anion requirement was demonstrated. Additionally, anaerobic or aerobic metabolism support a full hydroosmotic response equally well.


Isotope-release cytotoxicity assay with the use of indium-111: advantage over chromium-51 in long-term assays. The adaptation of indium-111-oxine (also known as 8-hydroxyquinoline) (111In Ox) chelate for long-term (18-48 hr) isotope-release assays of cell-mediated cytotoxicity (CMC) and its advantages over the use of 51 Cr are described. Labeling of DBA/2 P815 mastocytoma cells with 111InOx resulted in the incorporation of as many as a million counts per minute in 10(5) cells with no reduction in cell viability. 111InOx labeled both mouse and human tumor cells. 111InOx, like 51Cr, primarily labeled cytoplasmic constituents; up to 80% of the label existed in a releasable form. 111InOx was quantitatively released from labeled P815 in response to specifically sensitized C57BL/6 lymphocytes. The high labelling efficiency of 111InOx offered a significant advantage over 51Cr in 18- to 48-hour assays for CMC by reducing the counting error and thus making the assay more precise. Because of its higher labeling efficiency, 111InOx can be used in microcytotoxicity assays. 111InOx has the added advantage of a lower spontaneous release in culture than 51Cr. This feature of 111InOx also makes the calculation of specific isotope release more accurate than that achieved with 51Cr in long-term cytotoxic assays.


Blood microaggregates and ultrafilters.

Blood microaggregates and ultrafilters. The report presents three studies of post-traumatic pulmonary insufficiency (PTPI). In the first no significant pulmonary hemodynamic or ventilatory changes in severely shocked baboons resuscitated with shed fresh blood or stored blood were observed over 48 or 84 hours. Second, a post-mortem study of patients receving more than 5 units of blood within 24 hours of death showed sme microemboli in the lungs in about two thirds. Patients with multiple microthrombi had received an average of 20.6 units of blood; patients with some or no microemboli 15.5 and 6.3, respectively. Third in a review of the respiratory complications of 153 multiple-trauma patients, it was shown that the formerly severe problems with PTPI were now well managed clinically, that persistent respiratory failure was now occurring much later after injury, and occurred almost exclusively in patients with sepsis. Relation of the above data to previous reports in the literature led to the conclusion that the clinical significance of microaggregates in stored blood, if any, is low, and that ultrafiltration to remove microemboli only makes sense if it does not impede the rate of blood infusion and does not increase cost.


Effects of ethylenediaminetetraacetic acid-4 sodium solution on the surface of renal calculi and the uroepithelium. Tetrasodium ethylenediaminetetraacetic acid is one of the most effective agents that dissolve renal calculi, especially calculi containing calcium. To study the dissolving process of calculi in solutions, with or without trypsin, serial thin sections of renal calculi composed of calcium oxalate and/or phosphate were prepared and observed by polarizing microscopy. The effects of the solution on the surface of human renal calculi and rat uroepithelium also were studied by scanning electron microscopy. A remarkable difference in solubility was noted between the crystalline and the amorphous components in struvite calculi. Under the polarizing microscope the dissolving effect of 5 per cent tetrasodium ethylenediaminetetraacetic acid solution on struvite after 30 minutes of incubation was equivalent to the use of saline for 24 hours. The effect of trypsin was not apparent. However, under the scanning electron microscope trypsin accelerated the dissolving effect of the tetrasodium ethylenediaminetetraacetic acid solution in struvite and calcium oxalate calculi. The contradictory findings of the ineffectiveness of trypsin on the thin sections as opposed to the pronounced changes seen on the surface of the calculi could be attributed to the difference in the exposed area. The results suggest that trypsin does not seem to dissolve calculi but mainly facilitates the solution to permeate the calculi. The cellular arrangement of the uroepithelium was preserved in tetrasodium solution and in 0.04 per cent trypsin tetrasodium ethylenediaminetetraacetic acid solution.


Eye findings in the diagnosis of Fabry's disease. Patients with renal failure. Two patients underwent renal transplantation for what was thought to be glomerulonephritis and chronic pyelonephritis. The diagnosis of Fabry's disease was made as an incidental finding during an ophthalmologic consultation for evaluation of blurred vision. These two cases illustrate the usefulness of an eye examination in the correct diagnosis in patients with the multisystem complaints of Fabry's disease. The correct diagnosis was extremely important in understanding the other manifestations of this disease in the affected patient and in the genetic counseling of the family.


The results of culture of gallbladder, bile and gall-stones. Bacterial cultures and sensitivity were done on gall-bladder, bile stones and gall-bladder wall in 35 patients and positive cultures were obtained in nine (25.7%). The most effective antibiotics were Gentamycin and Ampiclox. The incidence of post-operative complications appears to be significantly higher in patients with a positive culture.


Histalog gastric analysis.

Histalog gastric analysis. A histalog gastric analysis was done in 20 patients with gastroduodenal disorders and 12 subjects with non-ulcer dyspepsia who were used as controls. Due to overlap of secretory responses its use as a diagnostic tool is limited. About 46% patients with duodenal ulcer exceeded the upper limit of acid secretion of control subjects. Values for stimulated secretion in controls and the patients with gastric ulcer were the same. Endoscopy if possible is the investigation of choice for the diagnosis of gastroduodenal disorders and the secretory studies should be limited to patients under-going surgery for peptic ulceration.


The results of basal and post pentagastrin gastric secretory studies in apparently healthy subjects, a preliminary communication. Pentagastrin stimulated gastric secretory studies were carried out in 26 apparently healthy subjects. Each specimen of gastric juice was analysed for volume, pH, total acidity and major electrolytes. The data was analysed and compared with other published reports.


Carcinoma of gall bladder.

Carcinoma of gall bladder. A study of 118 newly diagnosed patients with gallbladder disease during a two year period showed that the frequency of gallbladder cancer was 8.4% and cholelithiasis played a significant role in its etiology. No remarkable differences were observed in the clinical and biochemical findings between the benign and malignant disease of gallbladder.


Mycetoma caused by Nocardia asteroides.

Mycetoma caused by Nocardia asteroides. During the last 5 years six cases of mycetoma have been referred to the Razi Hospital, Thehran. Nocardia Asteroides was found to be the causative agent in one case only. Five of the patients were males, all being rice farmers, and one was a female. All of them belonged to Northern Iran which has a mediterranean climate. Treatment of the condition still remains unsatisfactory and ineffective and amputation may have to be performed as a last resort.


Therapeutic effect of immunization with OEP, protease toxoid and elastase toxoid on corneal ulcers in mice due to Pseudomonas aeruginosa infection. The effectiveness of immunizing mice with protease toxoid (PT) or elastase toxoid (ET) on the corneal ulcerization due to either protease or elastase were investigated. Consequently, mice immunized with either PT or ET were protected from corneal ulcers experimentally induced by the homologous enzyme, either protease or elastase. Similarly, two kinds of rabbit immune sera, anti-PT and anti-ET, were found to prevent corneal ulcers by the homologous enzyme. Then, the therapeutic effects of vaccination with a single or mixed vaccine consisting of one, two or three components, i.e., PT, ET and/or the common antigen (OEP) of Pseudomonas aeruginosa, were examined on corneal ulcers in mice produced by live cultures of the bacteria. For the same purpose, administration of a single or combined rabbit immune sera against PT, ET and OEP was conducted. As a result, vaccination with the three-component-mixed vaccine or administration with rabbit immune sera combined with anti-PT and anti-ET sera in addition to anti-OEP serum, were found to be the most effective in preventing corneal ulceration as well as in treating corneal ulcers in mice.


Modern treatment of bronchial asthma.

Modern treatment of bronchial asthma. Various biochemical mediators and autonomic events lead to symptom-causing pathological changes in asthma attacks, that is, mucosal edema, mucous gland hypersecretion, and bronchial smooth muscle contraction. The discovery of alpha and beta adrenergic receptors, and the observation that cyclic AMP is the intracellular effector in cells stimulated by various hormones, led to a better understanding of the mechanism of action of medications of asthma. Emergence evaluation, in addition to history, physical findings, and physiological status, should include prior asthma history, physical findings, and physiological status, should include prior asthma history. Initial emergency therapy in patients with a history suggesting responsiveness to simple measures includes subcutaneous epinephrine, 0.2 to 0.5 mg, or terbutaline sulfate, 0.25 mg. Also, the patient may benefit from inhalation of an aerosolized bronchodilator. Patients who do not respond to initial treatment in three to four hours or who deteriorate, should be hospitalized. Hospitalized asthma patients should be constantly observed and monitored. The emergency treatment should be continued vigorously. Corticosteroid therapy should be started upon admission. The response rate to therapy in the hospitalized asthmatic is highly variable. Outpatient management involves patient education in the nature of asthma and in the fact that multiple drugs and frequent changes in therapy may be required to bring the symptoms under control.


[Clinical biochemical aspects of the prophylaxis and therapy of senile cataract with zinc aspartate (author's transl)]. In the aetiology and pathogenesis of senile cataract a disturbed glucose utilisation is found. This reflects a loss of activity of some key enzymes for glycolysis in the lens. These enzymes (ALD, G6PDH, HK, LDH, MDH and phosphofructokinase) are zinc metal enzymes. The decrease in activity of these enzymes can be compensated by the administration of specific cations. With zinc one can improve the impaired glucose metabolism occurring in old age. For the prophylaxis and therapy of senile cataract the prolonged administration of zinc aspartate is indicated. In the presence of magnesium deficiency magnesium salts should also be given. Cation eliminating exogenous or endogenous factors must be taken into consideration.


[Partial parenteral feeding in infants with special reference to the amino acid levels (author's transl)]. 12 infants aged between 6 weeks and several months were partially fed for a period of three days parenterally with a 6% amino acid solution adapted to breast milk as well as with an electrolyte solution with 10% glucose. The amino acid levels in the plasma were determined daily in the blood of fasting blood after an infusion period of 20 hours and after an interval of four hours. All the amino acids examined during the period of the study, revealed good homeostasis.


[The influence of disodium cromoglycate (DNCG) on the phagocytotic activity of neutrophile granulocytes (author's transl)]. Disodium cromoglycate (Intal) inhibits in vitro the phagocytotic activity of neutrophile granulocytes. In thirteen pairs of blinding analysed experiments it could be shown that the DNCG incubatet blood samples had a phagocytotic index below that of the control. The statistical significance of these findings was assessed by a variance analysis.


[Increased demethylation of aminopyrine under long-term treatment with anticonvulsive drugs (author's transl)]. Demethylation of aminopyrine was measured in 25 healthy controls and 19 epileptics on long-term treatment with anticonvulsants by the 14C-aminopyrine breath test. Compared to controls epileptics exhibited increased cumulative 14CO2-exhalation rates (88.7% at 30 min, 62.6% at 2 h and 24.8% at 8h) following ingestion of 2 mu Ci 14C-aminopyrine. The results suggest that long-term treatment with antiepileptic drugs results in increased demethylating function of the liver which can be easily detected by a simple breath analysis technique like decreased demethylation in chronic liver disease.


Circadian oscillations of body temperature in the marmoset, Callithrix jacchus. The rectal temperature of 8 marmosets was taken regularly throughout a 76 hour period. A pronounced circadian rhythm was detected: body temperature reached a maximum during the light phase and a minimum during the dark phase.


Effects of pelleting and sterilization of diet on 2 strains of rabbit coccidia. Both autoclaving and irradiation at levels currently used for other laboratory animal diets appear to be satisfactory for the elimination of 2 of the most robust and pathogenic species of rabbit coccidia. Pelleting by the usual methods, even at 68 degrees C, was found to be unsatisfactory in eliminating all oocysts from heavily infected diet.


Medical economics survey-methods study: cost-effectiveness of alternative survey strategies. The Medical Economics Survey-Methods Study was conducted under contract for the National Center for Health Statistics during 1975-76. The objective was to determine the cost-effectiveness of a variety of strategies under consideration for application in a national survey in 1977. Detailed data on health care utilization and expenditures were collected periodically from a panel of 691 Maryland households over a seven-month period. Issues tested through a random experimental design included the relative effectiveness of monthly versus bimonthly interviews and in-person versus telephone contact. Completeness and accuracy of information were determined through an extensive record check involving all providers and third party payers identified in the household check involving all providers and third party payers identified in the household survey. The results indicate that there are substantial deficiencies in the household reporting and that the household data for all types of medical services would benefit from the availability of record data in order to improve theri accuracy. The frequency of contact (monthly versus bimonthly) had little effect on the accuracy of reporting. Given the higher rate of attrition observed for the monthly procedures and their considerably greater cost, the bimonthly interval appears to offer advantages. In-person contact resulted in significantly better reporting compared with telephone contact in the Baltimore area, but no difference was found in more rural Washington County. Thus, the 10 per cent cost increase for in-person contact may well be justified by the improved accuracy of the data obtained for certain types of populations.


A computerized scheme for the preparation of parenteral nutrition regimes. An operational computerized system to aid clinicians in the design of total parenteral nutrition regimes is reported. Two computer programs are described. The 'Requirements' program uses patient data and the results of blood and fluid excretion analyses to calculate the nutritional requirements for a period of 24 hours. Accepted formulae are used in these calculations. The 'Bottle Selection' program fits an intravenous bottle regime to the patient requirements from a standard set of commercially available intravenous feeding solutions. The system has been well accepted by the clinical staff who are free to accept, modify or reject a regime. Financial savings have been noted by the pharmacy who can plan for a faster turnover of a smaller range of solutions. The programs have been used routinely on a programmable calculator and have been written in CORAL for a Ferranti Argus 700E computer.


Air caloric stimulation with tympanic membrane perforation. Warm air caloric stimulation in an ear with tympanic membrane perforation or mastoidectomy cavity often causes contralateral nystagmus. Secondary nystagmus is common. Our evidence with squirrel monkeys and patients indicates that the primary "inversion" results from endolymph cooling due to evaporative cooling due to evaporative cooling of the mucus lining the middle ear cavity, by the dry air stimulus. Disconjugate horizontal nystagmus was found in a patient with large eardrum perforation, after cold air caloric stimulation. The effect probably resulted from stimulation of the anterior or posterior vertical semicircular canal. Inverted or disconjugate caloric nystagmus after air stimulation is much more frequently due to tympanic membrane perforation, or moisture in the external ear, than to central nervous system disease.


[Juxtapapillary diverticula of the duodenum and bilio-pancreatic symptoms (author's transl)]. Juxtapapillary diverticula of the duodenum are more frequent than has been assumed up to now. Among 2100 patients undergoing duodenoscopy 135 duodenal diverticula have been found. These diverticula may cause symptoms of diverculitis as well as of bilio-pancreatic disease. Such diverticula ought to be treated by surgery. In our department diverticula are turned inward, or a papillo-diverticulotomy is done depending upon the localization of the diverticula.


Cerebral prostaglandin synthesis during the dietary and pathological stresses of essential fatty acid deficiency and experimental allergic encephalomyelitis. Rats of the Lewis strain were fed diets adequate or deficient in essential fatty acids (EFA). At 70-80 days of age experimental allergic encephalomyelitis (EAE) was induced using adjuvants containing either Mycobacterium butyricum or Mycobacterium tuberculois H37Ra. When the former Mycobacterium was used, the incidence of EAE was greater in the EFA-deficient than in EFA-adequate controls; but when the rats challenged with M. tuberculosis, the incidence of the disease was the same in both dietary groups. Brain slices from EFA-deficient rats had a marginally depressed synthesis of prostaglandin F (PGF) compared to that of controls. Immunochallenge with adjuvant alone or adjuvant plus antigen tended to depress further PGF synthesis by brain slices from EFA-deficient rats and significantly depressed synthesis by slices from rats receiving adequate EFA. Whether or not rats were paralyzed had no effect on PGF synthesis when the diet was adequate in EFA, but a significant difference was seen in the EFA-deficient group. The results indicate a possible role for PGF synthesis in the degree of susceptibility of the rats to EAE under different dietary regimens.


Effects of acute endotoxemia and glucose administration on circulating leukocyte populations in normal and diabetic subjects. Effects of intravenous endotoxin and glucose administration on circulating leukocyte populations were compared in seven normal subjects and seven patients with juvenile-onset diabetes by means of automated cytochemical differential counting to quantitate each cell type. Both groups had comparable control cell counts that were unaffected by glucose tolerance testing but altered significantly by endotoxin. Different patterns of response to endotoxin were observed for different circulating cell types. The response of diabetics was parallel to that of normals but showed lower neutrophil and monocyte rebound, longer lasting depression of lymphocytes and eosinophils, and greater rebound of basophils on the day following endotoxin exposure. Characterization of distinctive normal response patterns of circulating leukocyte populations to endotoxin and comparison with responses in diabetes revealed abnormalities under conditions of stress that may impair the diabetic's ability to cope with acute infection.


The blue-green algae in nuclear power plant cooling water. A study was undertaken to determine the effects of passage through the cooling system of the Zion Nuclear Plant, Illinois, U.S.A., on the blue-green algae present in lake water. The blue-green algae were isolated by means of membrane filtration, enumerated and identified to genera. As a result of cooling system passage there was a definite reduction in the numbers of blue-green algae. This reduction appeared to be due to mechanical, as well as elevated temperature, damage. A few genera showed only a slight change in number.


Divalent cation mobility throughout exponential growth and sporulation of Bacillus megaterium. Each of the five elements considered was taken up by Bacillus megaterium during exponential growth. Initial Mg and Mn uptake was rapid and ended by mid-log. For Ca, Fe, and Zn, uptake continued throughout exponential growth. Elements were released from the cells immediately following initial uptake. For Mn, egression continued to t2, with release of 36% of total accumulated. Secondary uptake followed immediately and continued through stage V. Magnesium egression continued to t1 with release of 33% accumulated. Secondary uptake began by t5 (stage IV) and continued slowly through sporulation. Calcium egression ceased by t4 with release of 25% total accumulated. Secondary uptake began by t6 (stage V) and continued until depleted. Zinc egression stopped by t5 with release of 34% accumulated with some secondary uptake by stage V. Iron egression terminated at t4 with release of 59% of total accumulated. This was followed by secondary uptake after t12 (stage VI).


A separation of staphylococci and micrococci based on serological reactivity with antiserum specific for polyglycerophosphate. A serological reaction with the antiserum against heterophile polyglycerophosphate (PGP) was evaluated for genus level differentiation among strains of Staphylococcus and Micrococcus-Sarcina spp.. Hot saline extracts from whole cells of Staphylococcus spp. strongly reacted with the PGP antiserum, whereas those of Micrococcus-Sarcina spp. did not. Likewise, phenol-water extracts from whole cells of Micrococcus-Sarcina spp. were not reactive with the PGP antiserum, although the extracts of staphylococcal cells again gave a strong reaction with the antiserum. This study indicates that extracts from Micrococcus-Sarcina spp. have no antigen reactive with the PGP antiserum and can thus be differentiated from extracts of Staphylococcus spp. which react strongly with the PGP antiserum.


Inhibition by ammonium ion of germination of unactivated spores of Bacillus cereus T induced by l-alanine and inosine. Studies were carried out on the inhibitory effect of NH4+ on germination of spores of Bacillus cereus T induced by L-alanine and inosine. Kinetic analysis showed that NH4+ inhibited the germination competitively. Its inhibitory effect was greater when the unactivated spores had been preincubated with L-alanine. NH4+ did not inhibit the response of unactivated spores to L-alanine during preincubation. These results suggest that L-alanine sensitizes the spores to the inhibitory effect of NH4+.


[Lytic enzymes produced by Actinomyces levoris]. The composition of the enzyme complex produced by Actinomyces levoris which exhibits lytic action on the cell walls of Streptococcus lactis was studied by gel filtration on Sephadex G-25, ion exchange chromatography and isoelectric focusing. The complex was found to contain at least seven enzymes having different isoelectric points and substrate specificity. The enzymes were divided into three groups. (1) The enzyme with pI 10.2 seems to be a specific glucosidase; it possesses a high activity of the cell wall lysis and lacks a proteolytic activity. (2) The enzymes with pI 9.2 and 9.0 are, apparently, specific lytic proteases; they display both lytic and proteolytic activities. (3) The enzymes with pI 10.0, 9.5, 5.7 and 4.2 are, presumably, non-specific lytic proteases; they have a low lytic activity and a high proteolytic activity.


[Acylase activity of mycophilic fungi].

[Acylase activity of mycophilic fungi]. The acylase activity was studied with 65 cultures of mycophilic fungi belonging to 56 species and 33 genera. Among these: 9 species displayed the acylase activity toward ampicillin; 8 species, toward phenoxymethylpenicillin; 6 species, toward benzylpenicillin; and 21 species manifested the complex activity. Many of the active species belonged to the bionecrotrophic group of mycophilic fungi, the number of necrotrophic fungi was less, while that of biotrophs and saprotrophs was even lower.