Can J Microbiol, 1980 Apr, 26(4), 556 - 9 The atypical cell wall composition of Thermomicrobium roseum; Merkel GJ et al.; The cell wall of Thermomicrobium roseum, a Gram-negative, obligately thermophilic bacterium, has a composition unlike other Gram-negative thermophilic organisms . The purified cell wall was composed predominantly of a protein with a monomeric molecular weight of 75 000 . The amino acid composition of this protein revealed high concentrations of proline, glutamic acid, glycine, and alanine. Appl Environ Microbiol, 1980 Apr, 39(4), 840 - 4 Microbial prevalence in domestic humidifiers; Burge HA et al.; The prevalence of viable thermophilic bacteria and actinomycetes and mesophilic fungi was examined in 145 samples from 110 domestic humidifiers . A total of 72 and 43% of furnace and console humidifier samples, respectively, contained viable thermophilic bacteria, whereas 60 and 72% of these samples produced mesophilic fungal growth . Thermophilic actinomycetes were recovered from seven humidifier samples . Efforts to detect thermophilic actinomycete antigens in 15 humidifier fluid samples were not successful . Antifoulants added to humidifier fluid reservoirs had no apparent effect on microbial frequency . Airborne microbial recoveries did not reflect patterns of humidifier contamination with respect to either kinds or numbers of microorganisms in 20 homes in which volumetric air samples were obtained during humidifier operation. Proc Natl Acad Sci U S A, 1980 Apr, 77(4), 1922 - 6 Thermally induced biosynthesis of 2'-O-methylguanosine in tRNA from an extreme thermophile, Thermus thermophilus HB27; Kumagai I et al.; The contents of 2'-O-methylguanosine and 1-methyladenosine in unfractionated tRNA obtained from Thermus thermophilus HB27 were found to increase significantly when the bacterium was grown at a higher temperature (80 degrees C) . S-Adenosyl-L-methionine-dependent tRNA (guanosine-2')-methyltransferase (EC 2.1.1.34) and tRNA (adenine-1)-methyltransferase (EC 2.1.1.36) were detected in a cell-free extract of the thermophile, and both of them were partially purified . tRNA (guanosine-2')-methyltransferase specifically catalyzed the methylation of the guanylate residue at position 19 from the 5' end of Escherichia coli tRNAMetf . The amounts of these methyltransferases in the cells and their thermal characteristics seemed to be independent of the growth temperature of the bacterial cells from which the enzymes were extracted . It was inferred that the temperature dependence of the methylation process in vivo is accounted for, not by temperature dependence of enzyme formation, but by that of the enzyme activity. Mycopathologia, 1980 Mar 31, 70(3), 169 - 79 Thermophillic and thermotolerant fungi isolated from the thermal effluent of nuclear power generating reactors: dispersal of human opportunistic and veterinary pathogenic fungi; Rippon JW et al.; Over a period of a year, samples of water, foam, microbial mat, soil and air were obtained from areas associated with the cooling canal of a nuclear power station . The seventeen sample sites included water in the cooling canal that was thermally enriched and soil and water adjacent to, upstream, downstream and at a distance from the generator . Air samples were taken at the plant and at various distances from the plant . Fifty-two species of thermotolerant and thermophilic fungi were isolated . Of these, eleven species are grouped as opportunistic Mucorales or opportunistic Aspergillus sp . One veterinary pathogen was also isolated (Dactylaria gallopava) . The opportunistic/pathogenic fungi were found primarily in the intake bay, the discharge bay and the cooling canal . Smaller numbers were obtained at both upstream and downstream locations . Soil samples near the cooling canal reflected an enrichment of thermophilous organisms, the previously mentioned opportunistic Mucorales and Aspergillus spp . Their numbers were found to be greater than that usually encountered in a mesophilic environment . However, air and soil samples taken at various distances from the power station indicated no greater abundance of these thermophilous fungi than would be expected from a thermal enriched environment . Our results indicate that there was no significant dissemination of thermophilous fungi from the thermal enriched effluents to the adjacent environment . These findings are consistent with the results of other investigators. Biochim Biophys Acta, 1980 Mar 7, 590(1), 24 - 33 Isolation and characterization of two rubredoxins from Clostridium thermoaceticum; Yang SS et al.; Two rubredoxins with similar molecular weights (about 6000) have been purified from Clostridium thermoaceticum, a thermophile and strict anaerobe . They exhibit minor differences in several properties like elution pattern from DEAE-cellulose column, isoelectric point, amino acid composition, absorption and EPR spectra and redox potential . Their chemical and physical properties are similar to those of other rubredoxins from anaerobic microorganisms. Can J Microbiol, 1980 Mar, 26(3), 377 - 84 Purification and properties of polygalacturonic acid trans-eliminase from Bacillus stearothermophilus; Karbassi A et al.; A strain of thermophilic bacteria, Bacillus stearothermophilus, with pectolytic activity has been isolated . It produced an endo-polygalacturonic acid trans-eliminase (endo-PATE, EC 4.2.2.2) extracellularly when grown at 65 degrees C on a pectic acid medium . The PATE was purified 62-fold by the rapid affinity chromatographic method on a Sepharose-polygalacturonamide linked matrix . The absorbed PATE was eluted from the column with a continuous gradient of 0-10(-3)M ethylenediaminetetraacetic acid (EDTA) in phosphate buffer at pH 7.6 . The endo-PATE of this organism was much more heat stable than similar enzymes from the mesophilic Bacillus polymyxa and the thermotolerant Bacillus pumilus . The maximum activity of the enzyme occurred at 70 degrees C . With pectic acid as the substrate, the endo-PATE had an optimal pH of 9.0, the highest optimal pH compared with those of similar enzymes from other species of the genus . The molecular weight of the endo-PATE, as determined by chromatography on a Sephadex G-100 gel column, was 24 000. J Gen Microbiol, 1980 Mar, 117(1), 201 - 10 Phenotypic and genotypic characterization of some thermophilic species of Bacillus; Sharp RJ et al.; Some thermophilic species of Bacillus were characterized using biochemical tests, antibiotic sensitivity, bacteriocin and bacteriophage sensitivity, esterase patterns, DNA hybridization and % G+C content . The three caldo-active strains of bacilli isolated by Heinen (1971) were compared with strains of B . stearothermophilus . Eight of the strains examined showed characteristics which enabled them to be placed in the three main taxonomic groups suggested by Walker & Wolf (1971) . Two strains showed characteristics of both groups 1 and 3 . Examination of genotype data showed taxonomic groupings which differed from those based on phenotypic characterization. Am J Public Health, 1980 Mar, 70(3), 241 - 5 Respiratory symptoms and lung function in a sample of Vermont dairymen and industrial workers; Babbott FL Jr et al.; This study reviews the respiratory status of a sample of Vermont male dairy farmers, and a comparison group from industry, matched for age, sex and smoking . Survey instruments included a standardized questionnaire and simple pulmonary function tests . In general, past and present smokers had more respiratory symptoms than never-smokers; and farmers, in all smoking categories, reported symptoms with greater frequency than did their counterparts from industry . Forced vital capacity (FVC) tended to be lower among men with a history of smoking but, within each smoking category, dairymen and factory workers had very similar FVCs . Farmers who had never smoked or who were current cigarette users had lower FEV1/FVC (forced expiratory volume at one second/forced vital capacity) ratios than their controls from industry . Sixteen diarymen demonstrated precipitins to either Micropolyspora faeni (13) or Thermoactinomyces vulgaris, (3), but only one reported a constellation of symptoms compatible with farmer's lung disease . The estimated prevalence of antibodies to thermophilic actinomyces in this farm population was approximately 10 per cent . Although sample sizes were limited, dairymen from small farms tended to be older, have more respiratory symptoms, less satisfactory pulmonary function, and more serologic evidence of exposure to farmer's lung antigens than their counterparts from large farms. J Biochem (Tokyo), 1980 Mar, 87(3), 745 - 51 Bacillus thermoglucosidius alpha-glucosidase . Temperature dependence of activity and stability; Suzuki Y et al.; A p-nitrophenyl-alpha-D-glucopyranoside-hydrolyzing alpha-glucosidase from an obligate thermophile, Bacillus thermoglucosidius KP 1006, gave a triphasic relationship at pH 6.8 in the van't Hoff plot of Km, in the Arrhenius plot of the first order rate constant of inactivation with 0.1% sodium dodecyl sulfate, and in the logarithmic plot of the maximal fluorescence intensity at 346 nm versus reciprocal of temperature . The respective plots exhibited two breaks at 40 and 61 degrees C, 43 and 62 degrees C, and 40 and 61 degrees C . However, the Arrhenius plot of the molecular activity at pH 6.8 had a single discontinuity at 64 degrees C . These findings, together with thermodynamic quantities for the enzyme, suggest that the thermal conformation changes in the enzyme protein occur around 40--43 degrees C and 61--64 degrees C . The Arrhenius plot of the rate constant of heat inactivation at pH 6.8 was bent at 73 degrees C . Thermodynamic data indicate that the enzyme is transformed from a heat stable form into a heat unstable form at 73 degrees C with temperature elevation . The critical points localized near the minimum, optimum, and maximum temperatures (40, 60, and 72 degrees C) of the cell growth, respectively. Appl Environ Microbiol, 1980 Mar, 39(3), 584 - 7 Effect of temperature on mineralization by heterotrophic bacteria; Tison DL et al.; When pure cultures of the bacteria Pseudomonas fluorescens (a psychrotroph), Escherichia coli (a mesophile), and SRL 261 (a thermophile) were shifted away from temperatures to which they were adapted, the percentage of substrate mineralized increased (percent mineralized = {substrate respired to CO2}/substrate respired to CO2 + substrate incorporated into biomass} X 100) . The increase in the percent mineralized was larger for larger temperature shifts . Similar responses were observed when natural heterotrophic bacterial populations from sediments of Lake George, N.Y., and a thermophilic algal-bacterial mat community at the Savannah River Plant, Aiken, S.C., were subjected to temperature shifts . These results suggest that an increase in the percent mineralized may be an indication of thermal stress in bacterial populations. Mikrobiologiia, 1980 Mar-Apr, 49(2), 197 - 201 {Effect of temperature and pH on the ribulose-1,5-diphosphate carboxylase activity of the thermophilic hydrogen bacterium, Pseudomonas thermophila}; Romanova AK et al.; The activity of ribulose 1,5-diphosphate (RDP) carboxylase was found in the soluble fraction of the cytoplasm from sonicated Pseudomonas thermophila K-2 cells . The enzyme is relatively thermolabile and completely loses its activity at 80 degrees C . The activity of RDP carboxylase at 60 degrees C increases by 40% during the first 10 min of heating in the presence of Mg2+ ions, bicarbonate and dithiothreitol, and again decreases if the enzyme is heated over 20 min . The optimum temperature of the enzyme is 50--55 degrees C . The specific activity of the enzyme in fresh preparations under these conditions reaches 0.22 unit per 1 mg of protein in the extract . The calculated value of the activation energy for RDP carboxylase is 6.4 kcal-mole-1, but 11.6 kcal-mole-1 in frozen preparations . The optimal pH is 7.0--7.3 depending on the buffer . The temperature optimum for the enzyme action does not depend on pH within the range of 7.3 to 8.8 . Therefore, RDP carboxylase of Ps . thermophila K-2 differs from RDP carboxylases of mesophilic cultures studied earlier by a higher susceptibility to a decrease in temperature (the enzyme activity is negligible at 30 degrees C), by a lower value of the activation energy at suboptimal temperatures, and by a lower pH optimum of the enzyme action. Differentiation, 1980 Feb, 16(1), 61 - 9 Perturbance analysis of nuclear determination in Tetrahymena: analysis of mating type frequency variations with reference to binary-switch models; Portnoy S et al.; To test a 3-switch binary element model of mating type determination all possible assignments of the seven mating types of Tetrahymena thermophila to compound switch states have been examined, using a set of 77 experimentally obtained frequency patterns as a base . None of the assignments gives a satisfactory agreement with all the data . The result is similar whether the eighth switch state is considered forbidden or redundant . Moreover, further explorations of models in which the switches are not independent (as originally proposed), but coupled also fail to reveal a satisfactory agreement . Finally some 4-switch models are examined, again without discovering a satisfactory fit, though some evidence of structural relationship among the mating types is provided . The system is more complex than was assumed by the original models . To carry out this analysis certain statistical problems of a more general interest required solution . These include the management of samples from multinomial populations and the provision of efficient estimators for models of this sort. Differentiation, 1980 Feb, 16(1), 49 - 60 Perturbance analysis of nuclear determination in Tetrahymena: effects of nutrition, cell extracts, and CaCl2 on A/B hybrids; Nanney DL et al.; Mating type frequencies were ascertained among the progeny of crosses of strains A x B, Tetrahymena thermophila under a number of different circumstances . The frequencies are different if the parents are severely starved than if they are well-fed at the time of conjugation; severe starvation of the progeny before the first post-zygotic division has an effect similar to that of starving the parents . Mating type frequencies may also be modified by isolating conjugating pairs into cell extracts before the new macronuclei begin to develop; the changes do not appear to be related in a meaningful way to the mating type of the cells used as a source of the cell extracts . A third means of changing the mating type frequencies involves the exposure of conjugating pairs to CaCl2 solutions . Finally, changed frequency patterns may appear "spontaneously", and reflect either some as yet unsuspected environmental variable, or else an intrinsic metastable state that conditions the probabilities of mating type fixation . With the exception of the starvation effects, the pattern variations seem to fall into two groups . No satisfactory mechanism to account for these results is yet available. Cell, 1980 Feb, 19(2), 331 - 8 In vitro splicing of the ribosomal RNA precursor in nuclei of Tetrahymena; Zaug AJ et al.; The macronuclear rRNA genes of Tetrahymena thermophila contain a 0.4 kb intervening sequence . In this paper we present evidence that the excision of the intervening sequence from pre-rRNA occurs in vitro in isolated T . thermophila nuclei . The transcription-processing system includes alpha-amanitin to inhibit non-rRNA synthesis and aurintricarboxylic acid to inhibit endogenous nucleases . A discrete 0.4 kb RNA comprises up to 6% of the RNA synthesized in this system . Southern hybridization with restriction fragments of the rDNA (rRNA genes) shows that the 0.4 kb RNA contains the intervening sequence . The size of the 0.4 kb RNA, 410-425 nucleotides, is the size predicted if the entire intervening sequence were excised as a single linear molecule . The 0.4 kb RNA accumulates post-transcriptionally and appears to be stable in vitro . T . pigmentosa strain 6UM, whose rDNA has an intervening sequence of the same size and location as that of T . thermophila, also produces a 0.4 kb RNA in vitro; T . pigmentosa strain 8ALP, with not rDNA intervening sequence, produces no such RNA . The measurement of the accumulation of the excised intervening sequence is a convenient assay for the pre-rRNA splicing activity and a means for characterizing some of the details of the reaction . The resistance of the splicing acticity to concentrations of aurintricarboxylic acid that inhibit other endogenous nucleases should be useful in assaying the splicing enzyme during purification. Nucleic Acids Res, 1980 Jan 11, 8(1), 43 - 56 A spite specific endonuclease from thermus thermophilus 111, Tth111I; Shinomiya T et al.; A site specific endonuclease with novel specificity has been isolated from Thermus thermophilus strain 111 and named Tth111I . Tth111I cleaves lambda DNA into three fragments of 23.5, 25.7 and 50.8% of the total length, and ColE1 DNA into two fragments of nearly equal length . The sequences around Tth111I cleavage sites of ColE1 and lambda DNA were determined by the Maxam and Gilbert method and the two dimensional mapping method . The results suggest that Tth111I recognizes the DNA sequence (formula: see text) and cleaves the site as indicated by the arrows . Assuming that the first T.A pair in the sequence can be replaced for any base pair, the Tth111I recognition sequence has the symmetry with the two-fold axis as most type II restriction endonucleases do. Nutr Metab, 1980, 24(2), 65 - 75 {Bacteriological aspects of holding freshly prepared spinach at warm temperatures . Relation between bacterial count and nitrate reduction}; Bomar MT et al.; In sterilized spinach which was inoculated with bacteria, a reduction of NO3- to NO2-, due to their metabolic activity, was detectable, depending on the holding conditions . The reaction rate was highest in the presence of thermophilic bacteria . A lower reaction rate was altogether found with mesophilic bacteria which, depending on counts and holding time, may temporarily cause high nitrite contents . The reaction rate was lowest with cold-tolerant bacteria. Mikrobiologiia, 1980 Jan-Feb, 49(1), 74 - 7 {Thermostability and ultrastructure of submerged and aerial spores of Actinomyces chrysomallus}; Kuimova TF; Submerged and aerial spores of Actinomyces chrysomallus, strain 2703, producing the antitumour antibiotic chrysomallin occupy, in their thermoresistance, an intermediate position between spores of other mesophilic cultures and endospores of some thermophilic actinomycetes . These spores were heated in water at 90 and 95 degrees C for 10 min . The fraction of highly thermoresistant spores constituted less than 0.001% of all the spores . It was found for the first time in the submerged culture . The structure of the submerged and aerial spores was studied by electron microscopy and was shown to be similar to that of the exotype spores . Heating at high temperatures of the spores often resulted in the strong variability of the culture and in the appearance of proactinomycete-like variants. Chromosoma, 1980, 78(3), 313 - 25 Chromatin elimination and the genetic organisation of the macronucleus in Tetrahymena thermophila; Cleffmann G; In exponentially growing Tretrahymena thermophila the DNA content of the following structures was determined by cytophotometry: macronuclei of sister cells immediately after division; micronuclei; extranuclear chromatin in dividing cells and postdividers . Further, the development of macronuclear DNA amount in successive cell generations was determined . It was found that chromatin elimination is a frequent process reducing DNA content by about 4% per fission . This chromatin disappears within 20 min after division . The quantity of DNA extruded is highly variable and is different from the micronuclear DNA amount of multiples of it . The frequency of generations with two replication rounds as well as those without replication is estimated to be in the range of 2% each . These findings together with the qualitative difference between micro- and macronuclear DNAs suggest that the macronucleus of Tetrahymena is not entirely composed of complete genomes and that parts of the genetic material must be treated specifically for different sequences either during extrusion or during replication. Zentralbl Bakteriol Naturwiss, 1980, 135(1), 12 - 21 The effect of thermophilic actinomycetes isolated from compost and animal manure on some strains of Salmonella and Shigella; Makawi AA; Several samples of compost and animal manure were taken from different localities . Eleven isolates of thermophilic actinomycetes were obtained . The morphological, cultural, and physiological properties as well as temperature relation of these isolates were carried out in some detail in order to classify and identify them . The 11 isolates were identified as follows: Thermoactinomyces vulgaris (2 strains), Streptomyces thermofuscus (5 strains) and Thermoactinomyces sacchari (3 strains) . The antibacterial effect of these actinomycetes on the growth of some strains of Salmonella and Shigella was tested . Some of the Streptomyces had an effect on Salmonella . Thermoactinomyces vulgaris had no effect . Shigella strains were not affected. Folia Parasitol (Praha), 1980, 27(1), 63 - 9 A comparison of developmental dynamics of Dermacentor reticulatus (Fabr.) of different geographic origins and their affection by different microclimate; Daniel M et al.; Developmental dynamics of D . reticulatus ticks of different geographic origins (NE Poland and SW Slovakia) were investigated and compared . A long-term field experiment was conducted in South Moravia in the zone of thermophilic oak forest . Ticks were studied in two different biotopes: in an open grassy area and in deciduous forest . Differences between the tick populations collected in Poland and Slovakia were observed in the duration of developmental cycle and in the outcome of metamorphosis, depending on biotope . The tick development in the open grassy area was more rapid, but with greater losses . The total production of adults was higher in the population originating from Slovakia. Biochim Biophys Acta, 1980, 595(1), 140 - 5 Modulation of membrane fluidity in a fatty acid auxotrophe of Tetrahymena thermophila; Hill RJ; A mutant of Tetrahymena thermophila has been isolated which requires an unsaturated fatty acid for growth . Pellicles isolated from cultures supplemented with palmitoleic, elaidic, oleic, linoleic and gamma-linolenic acids show widely differing membrane fluidities, as measured by the polarisation of fluorescence technique . In contrast to the behaviour of the wild type organism, the changes in fluidity of the membrane, once induced by supplementation, are permanent . This mutant should prove extremely useful for studying structure-function relationships in the various membrane systems of Tetrahymena. Cell Motil, 1980, 1(1), 41 - 61 An analysis of spindle ultrastructure during anaphase of micronuclear division in Tetrahymena; LaFountain JR Jr et al.; Mitotic micronuclei were isolated from Tetrahymena thermophila and data on spindle ultrastructure were obtained from serial, transverse sections . Comparison of data from nuclei at meta- and early anaphase with data from nuclei at late anaphase showed that during anaphase, sister kinetochores move from the equator to the spindle poles, but kinetochore translocation occurs without any apparent change in either the number or length of kinetochore microtubules . This unprecedented result is ascribed significance with regard to the mechanism of kinetochore transport since there are only a limited number of ways that result could be achieved . The organization of the peripheral sheath changes during anaphase as evidenced by gaps in the sheath at late anaphase . Numerous kinetochore and non-kinetochore microtubules are located in polar regions of the spindle at late anaphase, whereas those regions contained only peripherally arranged microtubules at earlier stages . Tracking of individual kinetochore microtubules in late anaphase nuclei showed that some of them appeared to become incorporated into the peripheral sheath near the pole . At early and late anaphase, crossbridges connect adjacent microtubules throughout the spindle poleward to the kinetochores, as well as in the interzone. Z Allg Mikrobiol, 1980, 20(8), 513 - 6 Thermal resistance and viability of asexual spores of thermophilic fungi from composts; Ogundero VW et al.; The growth-temperature relations and thermal tolerance of the asexual spores of thermophilic fungi commonly found in composting plant materials were determined . The optimum temperature range for germination of the spores was 40 degrees - 50 degrees C . The spores of Humicola lanuginosa (GRIFFON and MAUBLANC) BUNCE were able to survive 60 min exposure to 68 degrees C while those of Mucor pusillus COONEY and EMERSON survived only 5 min exposure to 68 degrees C . A determination of the thermal death rates of the spores at 65 degrees C showed a rapid loss of viability of those of Torula thermophila COONEY and EMERSON . The thermal tolerance of the propagules of thermophilic fungi in relation to their special ecological niche is discussed. Zentralbl Bakteriol Naturwiss, 1980, 135(4), 275 - 89 {Physiological studies on thermophilic hydrocarbon-utilizing hyphae fungi (author's transl)}; Bemmann W et al.; Out of different substrates, thermophilic n-alkane-assimilating hyphae fungi were isolated and the strains of Aspergillus fumigatus and Mucor lusitanicus selected for physiological investigations . The growth curves, obtained in hydrocarbon and glucose at different temperatures, showed three phases: 1) initial growth phase, 2) linear growth phase, and 3) lysis phase . Maximum dry matter and maximum protein content occurred within a range of 35-40 degrees C . During the entire time of growth the DNA content per mg dry matter was almost the same, i.e., 0.1-0.2 per cent . The ascertained specific protease activity of Aspergillus fumigatus correlated with protein synthesis and protein decomposition of the biomass . The results of our investigations are being discussed. J Immunol Methods, 1980, 34(2), 167 - 75 Detection of proteinases in electrophoretograms of complex mixtures; Westergaard JL et al.; A simple, sensitive technique for detecting proteolytic enzyme zones on electrophoretograms by making contact print zymograms is described . The method is applicable to electrophoretograms prepared on a variety of support media, immunoelectrophoretograms or isoelectric focusing patterns on various media . The contact print zymograms are prepared by placing unfixed, unstained electrophoretograms in contact with a thin film of casein which has diffused into a layer of agarose supported by a hydrophilic polyester film . After staining the casein film with Coomassie blue, the proteolytic zones are detected as clear zones against a blue background . The method can detect as little as 9 ng of trypsin . The utility of the method is illustrated by detection of the proteinase enzymes in thermophilic actinomycete antigen preparations separated by polyacrylamide electrophoresis, crossed immunoelectrophoresis and isoelectric focusing on agarose and granulated dextran supports. Z Allg Mikrobiol, 1980, 20(2), 105 - 12 {A new selection method for obtaining mutants of Streptomyces glaucoachromogenes producing lambdamycin using the thermophilic phage-host system Tal/Thermoactinomyces vulgaris}; Fleck WF; In order to obtain high-producing mutants of the lambdamycin producer Streptomyces glaucoachromogenes IMET 31 118, a new selection technique using the thermophilic phage-host system Tal/Thermoactinomyces vulgaris, was developed . Lambdamycin is a yellow-green pigment antibiotic of the chromoglycoside type with antimicrobial, antiviral, cancerostatic, and ergotropic activity in vitro and in vivo . The physicochemical properties of lambdamycin resemble those of chartreusin . By means of the selection technique developed the determination of the concentration-dependent influence of lambdamycin on bacterial growth inhibition and Tal-phage development in the agar diffusion sphere of mutagen-treated colonies is possible without time and material-consuming pre-tests . A positive correlation exists between the biosynthetic capacity of mutants on solid media and in fermentation liquors . Using this new selection technique, the titres of lambdamycin in fermentation liquors of mutants could be increased by 10 to 20 fold in comparison to the wild type strain. J Biochem (Tokyo), 1980 Jan, 87(1), 1 - 13 Purification and thermal stability of several amino acid-specific tRNAs from an extreme thermophile, Thermus thermophilus HB8; Watanabe K et al.; Three species of methionine tRNAs and phenylalanine, tyrosine, and isoleucine tRNAs were purified from an extreme thermophile, Thermus thermophilus HB8 . Formylation studies of the three methionine tRNAs and their codon-specific binding activities to ribosomes showed that two of them (named tRNAf1Met and tRNAf2Met) were initiator tRNAs and the other (named tRNAmMet) was a non-initiator . The tRNAs from T . thermophilus all had melting temperatures of up to ten degrees higher than the corresponding species from E . coli . Most of the species also had slightly higher G+C contents than the corresponding species of E . coli, and each of them contained one mol each of the modified nucleosides, O2'-methylguanosine (Gm), 2-thioribothymidine (s2T), and 1-methyladenosine (m1A) . Their high melting temperatures could be explained by their high G+C contents and the presence of the modified nucleosides, espically s2T . Comparison of the melting temperatures of T . thermophilus tRNAf2Met with those of E . coli tRNAfMet and tRNAmMet at different magnesium concentrations showed that magnesium was also a factor in the thermostability of the thermophile tRNA. Mol Gen Genet, 1980, 180(1), 129 - 34 Galactokinase-deficient mutants of Tetrahymena thermophila: selection and characterization; Roberts CT Jr et al.; We have isolated a series of mutants of Tetrahymena thermophila which are resistant to inhibition of growth by the galactose analog, 2-deoxygalactose . These mutants were obtained after mutagenesis with nitrosoguanidine and the induction of cytogamy to permit the recovery of recessive mutations induced in the germline micronucleus . Resistance to 2-deoxygalactose is correlated with a decreased rate of growth in galactose minimal medium and greatly reduced levels of galactokinase . The resistant phenotype of the mutants is apparently due to the galactokinase deficiency, which prevents the accumulation of toxic phosphorylated metabolites of 2-deoxygalactose . Genetic analyses reveal that the 2-deoxygalactose resistance alleles segregate as single Mendelian loci . The galactokinase-deficient strains described here represent the first mutants in this organism for which the biochemical basis of the mutant phenotype is known . These mutants, as well as others isolated similarly, should be of value in the elucidation of the mechanisms governing galactokinase gene regulation and in improving techniques of selection for other recessive mutations in Tetrahymena. Biochim Biophys Acta, 1980, 613(1), 1 - 9 Physicochemical and catalytic properties of thermostable malate dehydrogenase from an extreme thermophile Thermus flavus AT-62; Iijima S et al.; Physicochemical and catalytic properties of thermostable malate dehydrogenase (L-malate:NAD+ oxidoreductase, EC 1.1.1.37), isolated from an extreme thermophile, Thermus flavus AT-62, were studied . The enzyme had a molecular weight of 67,000 and consisted of two subunits with an identical molecular weight . The helical content of the enzyme was estimated to be about 25% from the circular dichroism spectrum . The amino acid composition of the thermophilic enzyme was similar to that of its mesophilic counterparts . Titration with 5,5'-dithiobis(2-nitrobenzoic acid) showed that the enzyme contained only one sulfhydryl group per subunit . Substrate inhibition by oxaloacetate was observed . The inhibition decreased with increasing temperature, but was still significant at 60 degrees C . The enzyme was remarkably heat stable, without losing activity after incubation at 90 degrees C for 60 min . The melting temperature of the secondary structure of the enzyme was 96 degrees C. Eur J Biochem, 1980, 107(1), 57 - 65 Amino acid sequence of the proteolipid subunit of the proton-translocating ATPase complex from the thermophilic bacterium PS-3; Hoppe J et al.; The proteolipid subunit of the ATPase complex was identified in whole membranes of the thermophilic bacterium PS-3 by means of a covalent modification with the 14C-labelled inhibitor dicyclohexylcarbodiimide . The proteolipid could be purified from the membrane in free and carbodiimide-modified form by extraction with chloroform/methanol and subsequent carboxymethylcellulose chromatography in mixtures of chloroform/methanol/water . The complete amino acid sequence of the 72-residue polypeptide could be determined by automated solid-phase Edman degradation of the whole protein, and of fragments obtained after cleavage with cyanogen bromide and N-bromosuccinimide . Chemical cleavages and separations of the resulting fragments by gel chromatography were performed in 80% formic acid . The amino acid sequence shows a concentration of hydrophobic amino acids in two segments of about 25 residues at the amino-terminal and carboxy-terminal ends . The polar residues are clustered in the middle of the polypeptide chain . The bound {14C}dicyclohexylcarbodiimide label is recovered exclusively at position 56, which is occupied by a glutamyl residue . The proteolipid from PS-3 exhibits homology to the corresponding ATPase subunit from mitochondria . The carbodiimide-reactive glutamyl residue occurs at the position as in the mitochondrial proteins. Mikrobiologiia, 1980 Jan-Feb, 49(1), 151 - 5 {In vivo study of the growth characteristics of the microflora of Armenian yogurt}; Erzinkian LA et al.; The microflora from the Armenian yoghurt was studied during its growth on the optimal for the medium (skim milk) using objectives of the Peshkoff system . It was shown that certain organisms of Lactobacterium jogorti tended to branch at the early stages of their growth, and the cells elongated and formed complex coils and helices by the third-fourth day of growth . Compact aggregates of cocci having angular outlines appeared in the culture of Streptococcus thermophilus by the end of the second day of growth . Lysis of individual organisms, and occasionally zones of lysis, were found in the both cultures at certain stages of their growth . Cultures grown for 4-5 days were characterized by wide polymorphism. Nucleic Acids Symp Ser, 1980, (8), s181 - 4 A second site specific endonuclease from Thermus thermophilus 111,Tth111II; Shinomiya T et al.; A second site specific endonuclease with a novel specificity has been isolated from Thermus thermophilus strain 111 and named Tth111II . The enzyme is active at temperature up to 80 degrees C and requires Mg2+ or Mn2+ for activity . Tth111II cleaves phi X174RFDNA into 11 fragments . From the analysis of 5' terminal sequences of the phi X174RFDNA fragments produced by Tth111II action, it was concluded that Tth111II recognized the DNA sequence (See formula in text) and cleaved the sites as indicated by arrows. Ultrastruct Pathol, 1980 Jan-Mar, 1(1), 19 - 37 Langerhans cells and serum precipitating antibodies against fungal antigens in bronchioloalveolar cell carcinoma: possible association with pulmonary eosinophilic granuloma; Hammar SP et al.; In an ultrastructural study of 37 cases of bronchioloalveolar cell carcinoma (BAC), we identified seven cases (19%) in which Langerhans cells (LC) were closely associated with tumor cells . Serum precipitating antibodies against Aspergillus species and/or thermophilic actinomyces were present in five BAC patients whose tumors contained LC and in six patients whose tumors lacked LC . In a simultaneous study we identified marked atypical bronchiolar and alveolar lining cell hypertrophy and hyperplasma in pulmonary eosinophilic granuloma (PEG) . Our data plus current information suggesting that PEG is a form of hypersensitivity pneumonitis support our hypothesis that those cases of BAC in which LC are present may rise in localized or diffuse pulmonary scars caused by PEG. Can J Microbiol, 1980 Jan, 26(1), 58 - 63 Effect of prostaglandin E1-induced elevation of cyclic AMP on glucose repression in the lactic streptococci; Ratliff TL et al.; Cyclic adenosine 3', 5' -monophosphate (cAMP) activity was observed in Streptococcus lactis C2, Streptococcus cremoris C10, Streptococcus diacetlactis 18-16, and Streptococcus thermophilus C3 . In vitro assays of cell-free extracts obtaianed from S . lactis C2 showed that the cAMP-associated enzymes adenyl cyclase and phosphodiesterase were also present . In vitro experiments showed that prostaglandin E1 (PGE) stimulation of adenyl cyclase increased cAMP concentrations approximately fivefold, and in vivo studies showed that PGE treatment of S . lactis C2 increased intracellular cAMP concentrations twofold . Futhermore, PGE-induced elevation of intracellular cAMP levels was shown to prevent the repression of ss-D-phosphogalactoside galactohydrolase synthesis by glucose. Proc Natl Acad Sci U S A, 1980 Jan, 77(1), 147 - 51 Properties of a copper-containing cytochrome c1aa3 complex: a terminal oxidase of the extreme thermophile Thermus thermophilus HB8; Fee JA et al.; From the plasma membrane of Thermus thermophilus HB8 we have partially purified a detergent-solubilized complex of cytochromes a and c1 that actively catalyzes the transfer of electrons from ascorbate via a redox dye to oxygen . The complex is composed of two types of polypeptides, with molecular weights of approximately 55,000 and 33,000 . Quantitative analysis revealed the presence of heme a, heme c, and copper in a ratio of 2:1:2, with the heme a being present at 10 +/- 1.3 nmol/mg of protein . The heme c was shown to be associated with the molecular weight 33,000 peptide and is suggested to be of the c1 type . The optical and electron paramagnetic resonance properties of this complex were found to be similar to those of eukaryotic cytochrome oxidase, suggesting the following arrangement of chromophores: a magnetically isolated cytochrome c1 and an oxygen-reducing functional unit consisting of two heme a groups and two copper ions associated with one or more larger peptides. Biochemistry, 1979 Dec 11, 18(25), 5698 - 703 Thermal stability and protein structure; Argos P et al.; Amino acid sequences have been compared for thermophilic and mesophilic molecules of ferredoxin, glyceraldehyde-3-phosphate dehydrogenase, and lactate dehydrogenase . It is shown that Gly, Ser, Ser, Lys, and Asp in mesophiles are generally substituted by Ala, Ala, Thr, Arg, and Glu, respectively, in thermophiles . These exchanges suggest that thermal stability can be achieved by the addition of many small changes throughout the molecule without significant change in the backbone conformation . Their overall effect is primarily to increase internal and decrease external hydrophobicity as well as to favor helix stabilizing residues in helices . These substitutions minimize interruption of function or internal residue packing arrangements . Although the analysis has been confined to the above-mentioned molecules, the observed stabilizing principles may be more generally applicable. Biochemistry, 1979 Dec 11, 18(25), 5693 - 7 Denaturation of thermophilic ferricytochrome c-552 by acid, guanidine hydrochloride, and heat; Hon-nami K et al.; The denaturation of Thermus thermophilus cytochrome c-552 by acid, guanidine hydrochloride, and heat was studied by measuring the changes in absorption and circular dichroism . Cytochrome c-552 was remarkably resistant to acid; the pK of the transition from the low- to the high-spin form was roughly 0.3 . The effect of guanidine hydrochloride on the heme iron-methionine bond of Thermus and horse cytochromes c was also investigated; a comparison of the free-energy changes for the displacement of the bond indicated that the coordination in cytochrome c-552 is highly stable . The spectra of guanidine hydrochloride unfolded cytochrome c-552 were dependent on the pH; the titration curve showed the presence of a cooperative single transition of pK = 4.7, with a one-proton dissociation, suggesting the ionization of a histidine residue . In the presence of guanidine hydrochloride, the influence of the heat on the ligand bond in cytochrome c-552 was studied . The van't Hoff plots of the reaction were biphasic . The enthalpy changes in the higher temperature range were independent on the guanidine hydrochloride concentration, while those in the lower range were not. J Cell Sci, 1979 Dec, 40, 111 - 23 The regulation of amounts and proportions of genetic elements in the macronuclei of Tetrahymena thermophila strains of diverse karyotype; Seyfert HM et al.; Measurements of the micronuclear DNA content of clones with assumed different degrees of micronuclear ploidy confirmed the triploid nature of one clone . The micronuclear DNA content of a presumptive haploid clone was found to be slightly higher than expected, whereas one of two aneuploid clones had an unexpectedly low micronuclear DNA content . The macronuclear DNA content of cells with macronuclei which had developed from triploid, diploid and probably haploid and aneuploid macronuclear-Anlagen is very similar . Specifically, the smallest individual macronuclear DNA contents are consistently found within the same size class in all clones tested . Cell volumes, RNA and protein contents are alike in all clones tested . Only the growth rate and maximal density reached of one out of two aneuploid clones is reduced in comparison with the very similar other clones . The results are discussed with reference to the regulation of macronuclear DNA content, and to the compensation of gene-dosage. J Gen Microbiol, 1979 Dec, 115(2), 451 - 6 Mass selection of conditional mating mutants of Tetrahymena thermophila; Wolfe J; The mating reaction in Tetrahymena thermophila includes a starvation period and two distinct cell interactions, co-stimulation and cell pairing, before the cells are cytoplasmically joined as conjugants . A selection procedure for harvesting mutants unable to mate at a restrictive temperature has been developed . A conjugant pair consisting of one cycloheximide-resistant cell and one wild-type cell (cycloheximide-sensitive) was itself sensitive to the drug . By adding cycloheximide and nutrient medium to a cross made at the restrictive and grow . Repetition of the selection procedure enriched for cells unable to conjugate at the restrictive temperature . The selected cells were able to grow at 38 degrees C and could conjugate at 28 degrees C . This procedure may be narrowed to select specifically for cell interaction mutants. J Cell Physiol, 1979 Dec, 101(3), 503 - 13 Ribosome biosynthesis in Tetrahymena thermophila . IV . Regulation of ribosomal RNA synthesis in growing and growth arrested cells; Sutton CA et al.; Three parameters involved in the production of new ribosomal RNA (rRNA) were measured in Tetrahymena thermophilia: (i) the rate of synthesis of the rRNA precursor, (ii) the rate of processing of the RNA precursor and rRNA intermediates and (iii) the efficiency of utilization of the rRNA precursor in producing mature ribosomal RNA . These parameters were measured in cells in exponential growth and in cells starved in a dilute salt solution . Growing cells synthesize rRNA 20 times faster and process rRNA precursors and intermediates 10 to 15 times more rapidly than do starved cells . Both utilize their rRNA precursors with an efficiency of one in converting them to mature rRNA. J Biochem (Tokyo), 1979 Dec, 86(6), 1679 - 85 Occurrence of GTP cyclohydrolase I in Bacillus stearothermophilus; Suzuki Y et al.; A GTP cyclohydrolase which catalyzes the removal of carbon 8 of GTP as formic acid to yield a single pteridine compound occurs in an obligate thermophile Bacillus stearothermophilus ATCC 8005 . The enzyme was purified 5.5-fold . Its molecular weight and Stoke's radius were estimated as 105,000 and 45.3 A, respectively . The Km for GTP was 0.98 microM . The temperature and pH optima for activity were 60-65 degrees C and 8.0-8.4, respectively . No divalent cation was required for the reaction . The pteridine product was 3'-triphosphate of 2-amino-4-hydroxy-6-(D-erythro-1',2',3'-trihydroxypropyl)-7,8-dihydropteridine (dihydroneopterin triphosphate), identified by isolating its immediate derivative, 2',3'-cyclic phosphate of 2-amino-4-hydroxy-6-(D-erythro-1',2',3'-trihydroxypropyl)pteridine (neopterin cyclic phosphate) . The radioactive product from {8-14C}GTP agreed with 14C-formate . Molar ratio of formate release to pteridine formation was 1.0. Infect Immun, 1979 Dec, 26(3), 1057 - 64 Crossed immunoelectrophoretic analysis of two antigen extracts of Thermoactinomyces candidus; Hollick GE et al.; A pyridine extract antigen and a double-dialysis antigen (DDA) obtained from Thermoactinomyces candidus were analyzed by crossed immunoelectrophoresis . In addition, the heat lability, pronase sensitivity, and isolectric points of the components of the DDA were determined . By using antisera raised against crude pyridine extract antigen, two immunogenic components were resolved by crossed immunoelectrophoresis . A similar analysis of DDA using antisera raised against crude DDA revealed 15 immunogens . All but six components were heat labile, whereas pronase had little effect on the number of resolvable components . Intermediate gel crossed immunoelectrophoresis using antiserum raised to whole spores detected six immunogenic components, four of which were also detected by the anti-DDA serum . A total of 19 bands were obtained when the DDA was subjected to flatbed isoelectric focusing on polyacrylamide gels . The isoelectric points for the various components were found to range from 3.5 to 5.7 . Crossed immunoelectrophoresis using isoelectric focusing in the first dimension yielded at least 16 immunogenic components . Six components with isoelectric points falling in the range of 4.5 to 6.4 were found to be resistant to heat . A comparison with antigens obtained from other thermophilic actinomycetes is presented. J Biochem (Tokyo), 1979 Dec, 86(6), 1687 - 95 A double-alpha c-type cytochrome, cytochrome c-555, 549, from an extreme thermophile, Thermus thermophilus HB8; Hon-Nami K; A "double-alpha" c-type cytochrome, cytochrome c-555, 549, was isolated from the membrane fraction of an extreme thermophile, Thermus thermophilus HB8, and highly purified by chromatographies on DEAE-cellulose and Sephadex G-75 and by isoelectric focusing . The absorption maxima were at 554.8, 548.6, 522, and 417 nm in the reduced form, and at 528, 409, and 360 nm in the oxidized form . The double alpha-peak of this cytochrome was enhanced at liquid nitrogen temperature . The cytochrome contained one heme c group per protein molecule . The isoelectric point, midpoint redox potential and molecular weight were pH 4.0, +0.206 V and about 10,000, respectively . Cytochrome c-555, 549 is highly thermostable. Biophys Struct Mech, 1979 Dec, 6(1), 31 - 7 Thermal stability of soluble mitochondrial H+-ATPase; Kiladze AA et al.; ATPase melting has been studied by circular dichroism and differential scanning microcalorimetry . Decomposition of the alpha-helix of H+-ATPase (in which about 80% of the peptide groups of the enzyme are involved) following thermal treatment is shown to proceed gradually, beginning with room temperature . Effect of nucleotides upon melting is detected in the range of 20 degrees--40 degrees C . Above 40 degrees C, the pattern of thermal decomposition of the three-dimensional structure of H+-ATPase is independent of the nature of nucleotides present . Highly stable alpha-helical sites have been found in the enzyme molecule . Possible mechanism of formation of such sites is discussed, and the results obtained are compared with data on thermal stability of ATPase from thermophilic bacteria . Structural changes in the molecule following thermal treatment are compared with ATPase activity changes under similar experimental conditions. Biochim Biophys Acta, 1979 Nov 22, 565(1), 154 - 60 Stability of bacterial messenger RNA in mesophiles and thermophiles; Stenesh J et al.; The decay of {3H}uridine-labeled mRNA was measured in the mesophile, Bacillus licheniformis (grown at 37 degrees C and 46 degrees C), and in the thermophile, Bacillus stearothermophilus (grown at 46 degrees C and 55 degrees C) . For each organism, the half-life of the mRNA decreased as the growth temperature was increased . The stability index (half-life of mRNA/doubling time of cells), however, was remarkably constant for each organism regardless of the growth temperature . It is concluded that these results support the concept that kinetic considerations play a significant role in the explanation of thermophily. Mikrobiologiia, 1979 Nov-Dec, 48(6), 1113 - 7 {Distribution of thermophilic sulfate reducing bacteria in the oil-bearing strata of Apsheron and Western Siberia}; Rozanova EP et al.; Enrichment cultures of sulfate-reducing bacteria were obtained by inoculating the water of high-temperature gas and oil-bearing strata . A study of the morphology of these cells has shown that the thermophilic bacterium Desulfovibrio thermophilus occurs in the deposits of the Apsheron Peninsula while Desulfootomaculum nigrificans is found in the deposits of West Siberia . The former organism is involved in the accumulation of H2S in the Apsheron strata . The temperature of the strata is believed to be the factor which regulates the incidence of bacteria. Mech Ageing Dev, 1979 Nov, 11(4), 253 - 68 Germinal aging in Tetrahymena thermophila; Simon EM et al.; The manifestations of germinal aging in the ciliate Tetrahymena thermophila include death of the cells at conjugation and macronuclear retention in which the normal replacement of the old macronucleus by a new one fails to occur . Available data suggest that methods of routine maintenance that reduce the number of fissions may delay aging . Differences in breeding performance following maintenance for 1-5 years in axenic peptone broth vs . bacterized Cerophyl were not significant; those following maintenance at different temperatures were significant . The analysis of several hundred crosses is consistent with a random mutational basis for aging in the micronucleus and does not support the hypothesis of an age-correlated program in which the rate of deterioration increases with time . Following routine cultural maintenance for as long as 9-11 years, some lines show no deficiencies in their ability to produce viable progeny, and sublines of the same clone frequently differ significantly in their breeding performance . Moreover, breeding degeneration occurs at constant, but different, rates in different inbred strains. J Biochem (Tokyo), 1979 Nov, 86(5), 1427 - 32 Addition of short guanylyl blocks to oligonucleotide primers with a thermophilic polynucleotide phosphorylase . Its application to the synthesis of oligonucleotides containing guanylyl residues; Kikuchi Y et al.; Polynucleotide phosphorylase from Thermus thermophilus catalyzed the addition of short guanylyl blocks from GDP to the 3'-hydroxyl termini of oligonucleotide primers at low temperature in a simple reaction mixture . Polyguanylic acid formation was inhibited at 37 degrees C, but the addition of one or two guanylyl residues to oligonucleotide primers proceeded in high yields . The reaction was applied to the synthesis of oligonucleotides containing guanylyl residues at the 3'-end . Using (Ap)2A and (Up)2U as primers, (Ap)3G, (Ap)3GpG, and (Up)3G were synthesized in yields of 25--52% . (Ap)2GpG was synthesized from ApA and GDP in a yield of 13%. J Cell Physiol, 1979 Nov, 101(2), 349 - 58 Ribosome biosynthesis in Tetrahymena thermophila . III . Regulation of ribosomal RNA degradation in growing and growth arrested cells; Sutton CA et al.; We have measured the turnover rate of ribosomal RNA in exponentially growing Tetrahymena thermophila cells, cells entering the plateau phase of growth, and nutrient-deprived (starved) cells . Ribosomal RNA is stable in cells in early log phase growth but it begins to turnover as the cells begin a deceleratory growth phase prior to entering a plateau state . Likewise, rRNA in cells transferred from early log phase growth to a starvation medium begins to be degraded immediately upon starvation . In both cases the degradation of rRNA exhibits biphasic kinetics . A rapid initial exponential degradation with a half time of nine and one-half hours lasting for six hours is followed by a slower exponential degradation with a half-life of 35 hours . When starved cells are transferred to fresh growth medium turnover of rRNA ceases . The evidence presented suggests that the alteration in degradation rate is a regulated process which is most likely independent of the cell cycle. J Bacteriol, 1979 Nov, 140(2), 543 - 6 Protein turnover in the extreme thermophile Thermus aquaticus; Kenkel T et al.; Protein turnover in the extreme bacterial thermophile Thermus aquaticus was examined in exponential cultures at 75 degrees C . The relative amount of {3H}leucine incorporated into trichloroacetic acid-insoluble material was stable in pulse-chase experiments assayed over 2.5 h . The trichloroacetic acid-insoluble radioactive leucine was stable upon the addition of chloramphenicol, which blocks protein synthesis in T . aquaticus . The specific activity of a phosphate-repressible alkaline phosphatase, investigated in the presence of chloramphenicol, did not decrease . The addition of excess orthophosphate to cultures derepressed for the alkaline phosphatase did not show a marked effect on the specific activity over a 2-h period . On the basis of these four experiments, it does not appear that a high protein turnover rate is essential for the thermophily of T . aquaticus at 75 degrees C. J Gen Microbiol, 1979 Nov, 115(1), 253 - 4 Thermophilic and thermotolerant fungi in poultry droppings in Nigeria; Ogundero VW; Ten species of fungi were obtained from poultry droppings in Nigeria . Six of these are true thermophiles while the other four are thermotolerant . Aspergillus fumigatus Fresenius, Mucor pusillus Lindt and Thermoascus aurantiacus Stolk are known human pathogens . Except for M . pusillus, all the thermotolerant species had a higher occurrence at 45 degrees C while the thermophilic varieties were readily obtained at 50 degrees C. Eur J Biochem, 1979 Nov, 101(2), 317 - 24 5'-Methylthioadenosine phosphorylase from Caldariella acidophila . Purification and properties; Carteni'-Farina M et al.; The occurrence of 5'-methylthioadenosine phosphorylase in Caldariella acidophila, a thermophilic bacterium growing optimally at 87 degrees C, is reported . It represents the first example in prokaryotes of a phosphoryolytic cleavage of the thioether . The reaction products, purified by ion-exchange chromatography, have been identified as 5-methylthioribose-1-phosphate and adenine by several analytical procedures . The enzyme has been purified to homogeneity in 32% yield by using DEAE-cellulose and hydroxyapatite chromatography, gel filtration and isoelectric focusing . The enzyme shows a high degree of thermophilicity, its temperature optimum being at 93 degrees C; furthermore no loss of activity is observable after exposure for 1 h at 100 degrees C . The kinetic data indicate a sequential mechanism of the reaction . The apparent Km values are 0.095 mM for 5'-methylthioadenosine and 6.1 mM for phosphate . The specificity of the reaction is rather strict . Experiments performed with analogues of the substrate, i.e . 5'-methylthioinosine, 5'-dimethylthioadenosine sulfonium salt, 5'-n-butylthioadenosine, 5'-isobutylthioadenosine, 5'-isobutylthioinosine, adenosylhomocysteine, 5'-thioethanoladenosine, adenosine, indicate the relevance of the adenine amino group and the sulfur in thioether form in the binding to the enzyme protein. Biochemistry, 1979 Oct 16, 18(21), 4702 - 7 Lipoamide dehydrogenase from Malbranchea pulchella: isolation and characterization; McKay DJ et al.; Lipoamide dehydrogenase (EC 1.6.4.3) has been isolated from a total homogenate of frozen mycelium of the thermophilic fungus Malbranchea pulchella var . sulfurea by a three-step procedure involving ammonium sulfate fractionation, Procion Brilliant Blue M-R--Sepharose 4B chromatography, and hydroxylapatite chromatography . The second step is the key purification step with the Procion Brilliant Blue M-R dye acting as an affinity ligand for the enzyme . The purified enzyme gave a single protein band on polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate . The enzyme is a dimer of molecular weight 102 000, and each monomer of 51 000 molecular weight binds one molecule of flavin adenine dinucleotide . Other properties determined include a pH optimum of 8.2, a strong specificity for the substrates dihydrolipoamide and nicotinamide adenine dinucleotide, the apparent lack of multiple enzymic forms, the presence of diaphorase activity, and resistance to temperature denaturation up to 60 degrees C . The amino acid composition and absorption spectrum of the enzyme were also determined . The properties of lipoamide dehydrogenase from this source are very similar to those reported for the enzyme from serveral other sources. Biochim Biophys Acta, 1979 Oct 11, 570(2), 406 - 10 Regulatory characteristics of phosphoenolpyruvate carboxylase from the extreme thermophile, Thermus aquaticus; Sundaram TK et al.; Phosphoenolpyruvate carboxylase from the extremely thermophilic bacterium, Thermus aquaticus YT-1, exhibits a virtually absolute requirement for acetyl CoA and there is strong positive cooperativity in the interaction of this activator with the enzyme . Several tricarboxylic acid cycle intermediates inhibit the enzyme . These findings suggest an anaplerotic role for the enzyme and an allosteric modulation of its activity by acetyl CoA and tricarboxylic acid cycle intermediates. Biochim Biophys Acta, 1979 Oct 10, 548(1), 139 - 46 Exchange integral for a variety of tetranuclear ferredoxins; Blum H et al.; The temperature dependence of EPR spectra of oxidized {4Fe-4S}(-1,-2) ferredoxins (previously designated HiPIP) and a reduced {4Fe-4S}(-2,-3) ferredoxin have been analyzed so as to determine the energy of a low-lying excited electronic state . The values obtained were: Center S-3 from beef heart, 44 cm-1; Center S-3 from mung bean, 53 cm-1; the {4Fe-4S}(-1,-2) ferredoxin from Thermus thermophilus, 78 cm-1; Center N-2 of NADH ubiquinone reductase, 83 cm-1 . Increasing axial distortion in the EPR spectra of the {4Fe-4S}(-1,-2), ferrodoxins was associated with higher energy differences . Center N-2, a {Fe-4S}(-2,-3) iron-sulfur cluster does not fit this relationship. J Cell Sci, 1979 Oct, 39, 299 - 312 Mitochondrial associations with specific microtubular components of the cortex of Tetrahymena thermophila . I . Cortical patterning of mitochondria; Aufderheide K; Many of the mitochondria of Tetrahymena thermophila are localized in the cell cortex in regular, identifiable patterns . Two different mitochondrial patterns are seen; whether a cell expresses one or the other type apparently depends upon nutrient conditions in the culture and not upon other factors tested . Consistent associations between cortical mitochondria and certain of the cortical microtubular bands are seen at the light-microscopic level . Electron-microscopic observations confirm this and, furthermore, identify ultrastructural associations between cortical microtubules and the cortically located mitochondria . It appears that the cortical microtubular arrays serve as a guide for the localization (and thus patterning) of the cortical mitochondria. Cell, 1979 Oct, 18(2), 525 - 32 The intervening sequence in the 26S rRNA coding region of T . thermophila is transcribed within the largest stable precursor for rRNA; Din N et al.; We studied the transcription of the intervening sequence in the 26S rRNA coding region of the extrachromosomal rDNA molecules in the macronucleus of T . thermophila by hybridization of purified nuclear rRNA precursors or cytoplasmic 26S rRNA to purified native rDNA or specific rDNA restriction fragments . Examination of R loop hybrids in the electron microscope and analyses of S1-protected rDNA fragments in alkaline agarose gels showed that mature 26S rRNA, nuclear pre-26S rRNA and a fraction of the pre-rRNA molecules containing both the sequences for 17S and 26S rRNA all lack the region corresponding to the intervening sequence . The rest of the pre-rRNA molecules, however, hybridize in a colinear fashion to the whole coding region, and thus must contain the intervening sequence . We can conclude from these results that the intervening sequence is transcribed within the primary transcription product of the rDNA, and that the post-transcriptional removal of the intervening RNA sequence is a very early processing event in the organism. J Biochem (Tokyo), 1979 Oct, 86(4), 1055 - 65 Kietics of thermal unfolding and refolding of thermostable phosphoglycerate kinase; Nojima H et al.; The kinetics of denaturation by guanidine hydrochloride (GuHCl) of a thermostable phosphoglycerate kinase (PGK) extracted from Thermus thermophilus and of yeast PGK at neutral pH were studied by circular dichroism . Denaturation by GuHCl proceeded as a first-order reaction . The activation free energy of the denaturation reactions (delta Gf not identical to ) in the absence of GuHCl was estimated to be 32.7 kcal/mol for T . thermophilus PGK and 27.9 kcal/mol for yeast PGK (at 25 degrees C) . Measurements of the rate constants at various temperatures indicated that delta Gf not identical to has maximum values at 29 degrees C for T . thermophilus PGK and at 20 degrees C for yeast PGK, and that the temperature dependences of delta Gf not identical to, delta Hf not identical to, and delta Sf not identical to for T . thermophilus PGK are smaller than those of yeast PGK . Values of delta Sf not identical to for thermal denaturation for both PGK's are approximately 200 e.u. Proc Natl Acad Sci U S A, 1979 Oct, 76(10), 5051 - 5 Localization of transcribed regions on extrachromosomal ribosomal RNA genes of Tetrahymena thermophila by R-loop mapping; Cech TR et al.; R-loop hybridization and electron microscopy were used to map the RNA transcription products of the extrachromosomal rRNA genes of Tetrahymena thermophila . The mature 17S and 26S rRNAs and the nuclear 35S pre-rRNA and pre-26S rRNA were located with a precision of approximately 100 base pairs . A 370-base pair intervening sequence was found in the 26S coding region . It has the same size and relative location as that found in Tetrahymena pigmentosa {Wild, M.A . & Gall, J.G . (1979) Cell 16, 565-573} . One class of R-loop structures formed by nuclear pre-rRNA provided preliminary evidence for a primary transcript that contains the intervening sequence . The results suggested a processing scheme in which splicing of the intervening sequences is followed by a series of strand cleavages to give the mature 17S and 26S rRNAs . Analysis of the data also showed that RNA . DNA and DNA . DNA duplexes, when mounted for electron microscopy by the R-loop procedure, have the same length per base pair within 4%. Proc Natl Acad Sci U S A, 1979 Oct, 76(10), 4857 - 61 Micronuclei of Tetrahymena contain two types of histone H3; Allis CD et al.; Evidence is presented that micronuclei of Tetrahymena thermophila contain significant amounts of two types of histone H3 . One is indistinguishable from that found in macronuclei and the other is unique to micronuclei . The micronucleus-specific H3 has a slightly faster mobility than the common H3 in three different gel systems (both of these species were artifactually lost during procedures for histone preparation in previous studies) . Both micronuclear H3s appear to contain a single cysteine residue and are present in sucrose gradient-purified nucleosomes . Acid extracts from micronuclei also contain three prominent high molecular weight proteins that also were lost during previous procedures . These proteins are present in extracts from oligomers but are not observed in extracts from mononucleosomes, suggesting that they may be associated with linker regions between nucleosomes. J Gen Microbiol, 1979 Oct, 114(2), 401 - 8 Isolation and partial characterization of four plasmids from antibiotic-resistant thermophilic bacilli; Bingham AH et al.; Twenty-nine antibiotic-resistant isolates of thermophilic bacilli were examined for the presence of covalently closed circular duplex DNA molecules by agarose-gel electrophoresis and caesium chloride-ethidium bromide density gradient centrifugation . Five of the 29 strains tested contained covalently closed circular molecules . Two of the streptomycin-resistant strains contained the same two plasmids: pAB118A of molecular weight 4.9 X 10(6) (7.0 kilobases) and pAB118B of molecular weight 3.0 X 10(6) (4.3 kilobases) . Two of the tetracycline-resistant strains each contained a plasmid (pAB124) of molecular weight 2.9 X 10(6) (4.14 kilobases), while a third harboured a small plasmid (pAB128) of molecular weight 2.5 X 10(6) (3.57 kilobases) . These plasmids were digested with 19 different restriction endonucleases and the numbers of cleavage sites were determined . Transformation of Bacillus subtilis (168 (Trp-) with purified plasmid DNA indicated that pAB124 conferred tetracycline resistance on the host. J Biochem (Tokyo), 1979 Oct, 86(4), 893 - 905 Nucleotide sequence of formylmethionine tRNA from an extreme thermophile, Thermus thermophilus HB8; Watanabe K et al.; The nucleotide sequence of formylmethionine tRNA from an extreme thermophile, Thermus thermophilus HB8, was determined by a combination of classical methods using unlabeled samples to determine the sequences of the oligonucleotides of RNase T1 and RNase A digests and a rapid sequencing gel technique using 5'-32P labeled samples to determine overlapping sequences . Formylmethionine tRNA from T . thermophilus is composed of two species, tRNAf1Met and tRNAf2Met . Their nucleotide sequences are almost identical, and are also almost identical with that of E . coli tRNAfMet, except for slight modifications and replacements . Both species have modifications at three points which do not exist in E . coli tRNAfMet: 2'-O-methylation at G19, N-1-methylation at A59 and 2-thiolation at T55 . Moreover U51 in E . coli tRNAfMet is replaced by C51 in both species, so that a G-C pair is formed between this C51 and G65 . tRNAf2Met has a reversed G-C pair at positions 52 and 64 compared with those in tRNAf1Met and E . coli tRNAfMet . Other regions are mostly the same as those in all prokaryotic initiator tRNAs so far reported . The thermostability of these thermophile initiator tRNAs is discussed in relation to their unique modifications. J Biol Chem, 1979 Sep 25, 254(18), 8720 - 2 A new polyamine, thermospermine, 1,12-diamino-4,8-diazadodecane, from an extreme thermophile; Oshima T; A new polyamine has been extracted from an extreme thermophile, Thermus thermophilus, and its chemical structure was determined as 1,12-diamino-4,8-diazadodecane, NH2(CH2)3NH(CH2)3NH(CH2)4NH2, based on its proton NMR, 13C NMR, and mass spectra . A trivial name "thermospermine" is proposed for the new compound. Biochim Biophys Acta, 1979 Sep 21, 556(2), 265 - 77 Purification and partial characterization of a procaryotic glycoprotein from the plasma membrane of Thermoplasma acidophilum; Yang LL et al.; The obligate, thermophilic, acidophilic mycoplasma, Thermoplasma acidophilum, grows optimally at 56 degrees C and pH 2.0 . Its plasma membrane possessed 21--22 protein bands that were resolved by polyacrylamide gel electrophoresis . One major membrane protein, molecular weight 152 000, which stained for carbohydrate with periodic acid-Schiff reagent, accounted for 32% (w/w) of the total membrane proteins . It was isolated and further purified by concanavalin A affinity chromatography . The carbohydrate content amounted to less than 10% (w/w) compared to that of the entire glycoprotein . The carbohydrate moiety consisted mainly of mannose residues with branched alpha 1 leads to 2 linkages at the non-reducing ends of the glycopeptide as determined by permethylation followed by gas chromatography-mass spectrometry analysis . The reducing end was an N-glycosidic linkage between asparagine and N-acetylglucosamine . The amino acid composition of this glycoprotein showed 62 mol% hydrophobic residues, while the acidic amino acid content contributed 9 mol% more than that of the basic amino acids . The existence of membrane glycoproteins in the procaryotic, wall-less T . acidophilum may provide a protective coat for the plasma membrane . The stereochemistry and the conformation of the carbohydrate chains, in conjunciton with water turgor, may contribute to the rigidity of the membrane and the cation binding. J Cell Physiol, 1979 Sep, 100(3), 407 - 11 Peptidase activity in Tetrahymena; Zdanowski MK et al.; This report strongly suggests that two compartments in Tetrahymena thermophila contain peptidase activity: the cytoplasm and the outer cell surface . Determinations of amino acid concentrations in the extracellular medium upon incubation of cells with peptides suggest that the surface-bound peptidase activity hydrolyses di- and tri-phenylalanine equally fast on a molar basis . Growth experiments designed to characterize the in vivo peptidase specificities showed that both T . thermophila and T . pyriformis can use L-leucyl-L-leucine, but not L-leucyl-D-leucine as a leucine donor . These results are independent of whether the cells form food vacuoles or not. J Bacteriol, 1979 Sep, 139(3), 800 - 10 Isolation from soil and properties of the extreme thermophile Clostridium thermohydrosulfuricum; Wiegel J et al.; Thirteen strains of a strict anaerobic, extreme thermophilic bacterium were isolated from soil samples of moderate temperature, from a sewage plant in Georgia, and from hot springs in Utah and Wyoming . They were identified as strains of Clostridium thermohydrosulfuricum . The guanosine + cytosine content (moles percent) was 37.6 (determined by buoyant density) and 34.1 (determined by melting temperature) . All strains required a factor present in yeast extract or tryptone growth . Growth characteristics were as follows: a pH range of 5 to 9, with the optimum between 6.9 to 7.5, in a temperature range of 40 to 78 degrees C, with the optimum at 68 degrees C . The doubling time, when grown on glucose at temperature and pH optima, was 1.2 h . The main products of glucose fermentation were ethanol, lactate, acetate, CO2, and H2 . The fermentation was inhibited by H2 . Formation of spores occurred easily on glucose-agar medium or when cultures growing at temperatures above 65 degrees C were allowed to cool to temperature below 55 degrees C . C . thermohydrosulfuricum occurs widely distributed in the natural environment. Biochemistry, 1979 Aug 21, 18(17), 3647 - 53 Pyruvate carboxylase from a thermophilic Bacillus: some molecular characteristics; Libor S et al.; Analysis of the native enzyme and of the subunits produced upon its denaturation shows that pyruvate carboxylase from a thermophilic Bacillus is a tetramer with a molecular weight (mean value) of 558,000 and that the four polypeptide subunits are probably identical . The three functions (carboxyl carrier, carboxylation, and carboxyl transfer) in the pyruvate carboxylation reaction must therefore reside in this quarter-molecular polypeptide . The enzyme molecule contains four atoms of zinc and four molecules of D-biotin, and in the electron microscope the disposition of its four subunits presents a rhombic appearance . Reaction of the denatured enzyme with 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB) reveals 10 sulfhydryl groups/subunit . In the native enzyme less than one of these groups reacts with DTNB . By contrast, all of these groups (11/subunit) of the native chicken liver pyruvate carboxylase are accessible to DTNB . The thermophile enzyme is also more resistant to other sulfhydryl reagents and to denaturation under certain conditions than the avian enzyme. Biochim Biophys Acta, 1979 Aug 15, 569(2), 239 - 48 Ribulose 1,5-bisphosphate carboxylase from the thermophilic, acidophilic alga, Cyanidium caldarium (Geitler) . Purification, characterisation and thermostability of the enzyme; Ford TW; An an initial stage in the study of proteins from thermophilic algae, the enzyme ribulose 1,5-bisphosphate carboxylase 2-phospho-D-glycerate carboxylyase (dimerizing, EC 4.1.1.39) was purified 11-fold from the thermophilic alga Cyandium caldarium, with a 24% recovery . This purified enzyme appeared homogeneous on polyacrylamide gels and could be dissociated into two subunit types of molecular weights 55,000 and 14,900 . The optimal assay temperature was 42.5 degrees C, whilst enzyme purified from Chlorella spp . showed maximum activity at 35 degrees C . The thermostability of Cyanidium ribulose 1,5-bisphosphate carboxylase was considerably greater than that of the Chlorella enzyme, and the presence of Mg2+ and HCO-3 further enhanced this heat stability . A break in the Arrhenius plot occured at 20 degrees C for Chlorella ribulose 1,5-bisphosphate carboxylase and 36 degrees C for the enzyme from Cyanidium . It is suggested that the thermostability of Cyanidium ribulose 1,5-bisphosphate carboxylase is a result of an inherent stability of the enzyme molecule which permits efficient CO2 fixation at high temperatures but results in low activity in the mesophilic temperature range. Genetics, 1979 Aug, 92(4), 1079 - 92 An enrichment for temperature-sensitive mutants in Tetrahymena thermophila; Martindale DW et al.; The parameters for the killing of Tetrahymena by 5-bromodeoxyuridine(BUdR) and near-ultraviolet light have been determined . Significant preferential killing by UV of cells that have incorporated BUdR was obtained when the cells were irradiated in a nonnutrient buffer . UV alone was found to be toxic to cells irradiated in growth medium . Mutants defective in division at a restrictive temperature were isolated from mutagenized cultures that had been treated with BUdR and UV and from mutagenized cultures that had no such treatment . Results indicate that the number of temperature sensitive (ts) growth mutants can be increase five to six times using the BUdR/UV treatment . Data are presented that indicate differences in the frequency of occurrence of various types of ts mutants, with and without enrichment . A mutant that immediately stopped macromolecular synthesis and cell division upon being placed at the restrictive temperature was more resistant to BUdR/UV treatment than wild type by 1000-fold . Using the above techniques, BUdR-resistant mutants altered in the phosphorylation of thymidine have been isolated. Int J Pept Protein Res, 1979 Aug, 14(2), 99 - 106 Comparative temperature-stability properties of malate dehydrogenases from some thermophilic fungi; Wali AS et al.; Temperature-activity and temperature-stability relationships of malate dehydrogenases from eight thermophilic fungi were determined . Temperature optima for maximum activity of the enzymes varied between 50 degrees and 60 degrees and the Arrhenius plots were linear between 5 degrees and 50 degrees . The energies of activation ranged from 2.1 Kcal/mol for the enzyme from Sporotrichum thermophile to 9.1 Kcal/mol for that from Penicillium duponti . Heat inactivation kinetics at 50 degrees revealed heat lability of the enzyme from most of the thermophilic fungi . The t1/2's (min) were less than 10 for the enzymes from P . duponti, S . thermophile and Thermoascus aurantiacus; less than 30 for those from Chaetomium thermophile var . coprophile, H . lanuginosa and C . thermophile var . dissitum; and greater than 30 for those from Mucor pusillus and H . insolens . Salts of Na+, K+ and NH4+, and citrate protected the enzymes from H . lanuginosa, C . thermophile var . dissitum and M . pusillus against heat inactivation. J Protozool, 1979 Aug, 26(3), 519 - 24 Acid hydrolases and their release in food vacuole-less mutants of Tetrahymena thermophila; Silberstein GB; Mutants (NP1 and PSJ5) of Tetrahymena thermophila strains B and D 1968 exist that are unable to construct a functional oral apparatus and form food vacuoles at 37 C but which do so normally at 30 C . Food vacuole-less cells starved in dilute salt solution released similar amounts of acid phosphatase, beta-N-acetyl-glucosaminidase and alpha-glucosidase activity into the medium as wildtype cells during an 8-h period . Actively growing, food vacuole-less cells had approximately 50% less total protein, acid phosphatase, beta-N-acetyl-glucosaminidase, and alpha-glucosidase per cell than wildtype cells after 72-h growth . During this time food vacuole-less cells released significant amounts of the 3 acid hydrolases into the growth medium . For each hydrolase, the total activity released from growing, food vacuole-less cells was less, on a per cell basis, tahn the amount released from food vacuole formers . The proportion of the total activity secreted by the mutant and the wildtype cells was the same for acid phosphatase and beta-N-acetyl-glucosaminidase and somewhat lower for alpha-glucosidase . It is concluded that the release of a significant amount of acid hydrolase activity from Tetrahymena is independent of food vacuole formation and may be analogous to the secretory activity of other nonphagocytic eukaryotic cells. J Bioenerg Biomembr, 1979 Aug, 11(3-4), 39 - 78 Structure and function of H+-ATPase; Kagawa Y et al.; (1) Extensive studies on proton-translocating ATPase (H+-ATPase) revealed that H+-ATPase is an energy transforming device universally distributed in membranes of almost all kinds of cells . (2) Crystallization of the catalytic portion (F1) of H+-ATPase showed that F1 is a hexagonal molecule with a central hole . The diameter of F1 is about 90 A and its molecular weight is about 380,000 . (3) Use of thermophilic F1 permits the complete reconstitution of F1 from its five subunits (alpha, beta, gamma, delta, epsilon) and demonstration of the gate function of the gamma delta epsilon-complex, the catalytic function of beta (supported by alpha and gamma), and the H+-translocating functions of all five subunits . (4) Studies using purified thermostable F0 showed that F0 is an H+-channel portion of H+-ATPase . The direct measurement of H+-flux through F0, sequencing of DCCD-binding protein, and isolation of F1-binding protein are described . (5) The subunit stoichiometry of F1 may be alpha 3 beta 3 gamma delta epsilon . (6) Reconstitution of stable H+-ATPase-liposomes revealed that ATP is directly synthesized by the flow of H+ driven by an electrochemical potential gradient and that H+ is translocated by ATP hydrolysis . This rules out functions for all the hypothetical components that do not belong to H+-ATPase in H+-driven ATP synthesis . The roles of conformation change and other phenomena in ATP synthesis are also discussed. Biochim Biophys Acta, 1979 Jul 11, 569(1), 6 - 12 Activation of Thermus phosphofructokinase by monovalent cations; Stellwagen E et al.; The presence of the monovalent cations Tl+, NH+4, K+, Rb+ or Cs+, in decreasing order of potency, produce a marked equivalent increase in the specific enzyme activity of phosphofructokinase (ATP:D-fructose-6-phosphate 1-phosphotransferase, EC 2.7.1.11) purified from extreme thermophile, Thermus X-1 . By contrast, the monovalent cations Li+, Na+ or CH3NH+3 produce no detectable catalyitic activation at concentrations up to 100 mM . The relative potency of these cations suggests that each polypeptide chain in the tetrameric enzyme possesses a cationbinding site having tetragonal symmetry and that the protein ligands are principally hydroxyl or carboxylate oxygens . Only the enzyme-cation complex and not the enzyme by itself exhibits cooperativity with respect to the dependence of catalytic rate on the concentration of the substrate, fructose 6-phosphate . In the presence of subsaturating but not saturating concentrations of substrate, the catalytic activation produced by monovalent cations is also cooperative . Exclusion chromatographic measurements indicate that the enzyme remains tetrameric at catalytic concentrations in the presence or absence of an activating monovalent cation. Appl Environ Microbiol, 1979 Jul, 38(1), 166 - 8 Distribution of Aeromonas hydrophila in natural and man-made thermal effluents; Hazen TC et al.; Densities of Aeromonas hydrophila showed distinct thermal optima (25 to 35 degrees C) and thermal maxima (45 degrees C) when measured along thermal gradients created by geothermal and nuclear reactor effluents . Survival of A . hydrophila never exceeded 48 h at temperatures of greater than 45 degrees C . Thermophilic strains could not be isolated at any site. J Biochem (Tokyo), 1979 Jul, 86(1), 121 - 30 Effects of divalent cations on thermophilic inorganic pyrophosphatase; Hachimori A et al.; Divalent cations were shown to affect the structure and thermostability of thermophilic inorganic pyrophosphatase {pyrophosphate phosphohydrolase EC 3.6.1.1} purified from Bacillus stearothermophilus and thermophilic bacterium PS-3 . The properties of the enzymes from the two sources were found to be very similar . The enzymes were very unstable to heart in the absence of divalent cations, being inactivated gradually even at 40 degrees C . However, they became stable to heat denaturation in the presence of Mg2+, between pH 7.8 and 9.0 . Similar induced thermostability was detected when Mn2+, Co2+, Ca2+, Cd2+, and ZN2+ were added, though the latter three cations were not essential for enzyme activity . On adding divalent cations, the optical properties such as absorption spectra, fluorescence spectra, and circular dichroism (CD) were changed . Gel filtration and disc electrophoresis revealed that the molecular weight of both enzymes was 5.4 x 10(4) in Tris-SO4 buffer and 11 x 10(4) in Tris-HCL buffer, suggesting monomer-dimer transformation . In the presence of divalent cations in Tris-SO4 fuffer, the enzymes dimerized; this was confirmed by sedimentation velocity measurements . The enzymes in Tris-HCL buffer did not show thermostability unless divalent cations were added . The results in the present study indicate that binding of divalent cations to each enzyme caused some conformational change in the vicinity of aromatic amino acid residues leading to dimerization of the enzyme molecule so that it became thermostable . It was also suggested that histidyl residues play an important role in the thermostability induced by divalent cations on the basis of the pH dependencies of thermostability and CD spectra. Hoppe Seylers Z Physiol Chem, 1979 Jul, 360(7), 795 - 807 Structure and function of L-lactate dehydrogenases from thermophilic and mesophilic bacteria . I) Isolation and characterization of lactate dehydrogenases from thermophilic and mesophilic bacilli; Schar HP et al.; Lactate dehydrogenases from thermophilic bacilli (Bacillus stearothermophilus, Bacillus caldotenax) and from mesophilic bacilli (Bacillus X1, Bacillus subtilis) have been isolated by a two-step purification procedure . Only one type (LDH-P4) composed of four identical subunits (Mr 34 000 or 36 000) was found in each bacillus . The tetrameric enzymes were characterized with respect to thermostability, pH and temperature dependence of the pyruvate reduction and the L-lactate oxidation, substrate specificity, saturation kinetics (Km values of pyruvate, lactate, NAD, NADH), pyruvate and oxamate inhibition, and activation by fructose bisphosphate . The thermophilic and mesophilic enzymes differ characteristically in these parameters . Preliminary structural data (amino acid composition, comparative N-terminal sequence analysis) show the expected close phylogenetic relationship (high degree of sequence homology), but also typical differences between thermophilic and mesophilic dehydrogenases, a suitable basis for further comparative studies. Rev Bras Pesqui Med Biol, 1979 Jun, 12(2-3), 189 - 91 Thermophilic Campylobacter-associated diarrhoea in Rio de Janeiro; Ricciardi ID et al.; Thermophilic Campylobacter is reported as the agent associated with chronic and acute diarrhoea in Rio de Janeiro . Nine strains were isolated from 186 children with gastroenteritis . To our knowledge this is the first report of human Campylobacteriosis in Latin-America. J Gen Microbiol, 1979 Jun, 112(2), 357 - 64 The control of the synthesis of isocitrate lyase in a thermophilic bacillus; Griffiths MW; From a strain of Bacillus stearothermophilus, devoid of active pyruvate carboxylase, a mutant (NG-15) was selected that grew on acetate in the presence of glucose . This mutant differed from its parent organism in possessing high activities of isocitrate lyase when grown on all carbon sources tested except nutrient broth, in possessing unusually low activities of NADP+-dependent isocitrate dehydrogenase and in containing increased amounts of isocitrate . Revertants of mutant NG-15 which regained the ability to synthesize active pyruvate carboxylase also synthesized isocitrate lyase and isocitrate dehydrogenase to the same extent as the wild-type strain . These results suggest that the regulatory mechanism for the synthesis of isocitrate lyase in the thermophile may be different from that in mesophilic bacilli. J Biochem (Tokyo), 1979 Jun, 85(6), 1509 - 17 Purification and properties of phosphoglycerate kinase from Thermus thermophilus strain HB8; Nojima H et al.; (1) A glycolytic enzyme, phosphoglycerate kinase {EC 2.7.2.3}, was purified from cells of an extreme thermophile, Thermus thermophilus strain HB8 . The enzyme was resistant to heat, and no loss of activity was observed after incubation for 10--20 min at 79 degrees C . (2) Catalytic properties such as pH optimum (pH 6--8.5), kinetic parameters (Km=0.28 mM for ATP, 1.79 mM for glycerate 3-phosphate), substrate specificity and inhibitors of the enzyme were investigated and compared with those of phosphoglycerate kinase from other sources . (3) The enzyme protein consists of a single polypeptide chain of molecular weight 44,600 . The isoelectric point is 5.0 The amino acid composition of the enzyme was studied . The contents of ordered secondary structures were estimated to be 29% alpha-helix and 11% pleated sheet from the circular dichroic spectrum of the enzyme protein . (4) The fluorescence spectrum of the enzyme protein showed an emission maximum at 320 nm when excited at 280 nm . The quantum yield was 0.19 . Tryptophyl fluorescence was not quenched, in contrast to the fluorescence reported for yeast phosphoglycerate kinase. J Bacteriol, 1979 Jun, 138(3), 903 - 8 Use of 8-hydroxyquinoline to enrich for temperature-sensitive mutants of Tetrahymena; Martindale DW et al.; The effects of the chelating agent 8-hydroxyquinoline (Hq) on Tetrahymena thermophila were examined . Cell division was completely inhibited by 5 micrograms of Hq per ml . At this concentration deoxyribonucleic acid, ribonucleic acid, and protein syntheses were also completely and nonselectively inhibited . The inhibition was reversible after 6 h of Hq treatment . At concentrations above 20 micrograms/ml a 10,000-fold decrease in survival as seen after 2 h in the drug . The sensitivity of Tetrahymena to Hq was found to be dependent upon cell concentration, wild-type strain, medium, and length of time the culture is at 38 degrees C before Hq is added . Mutants of Tetrahymena that are unable to divide at the restrictive temperature, but which continue macromolecular synthesis, were found to be resistant to Hq treatment . Conditions were obtained in which more than a 1,000-fold difference in survival was seen between this class of mutant and the wild type . The effect of Hq on three other classes of temperature-sensitive mutants was examined, and the results are discussed. Sabouraudia, 1979 Jun, 17(2), 163 - 9 Antigenic relationships among thermophilic actinomycetes; Kurup VP et al.; Antigens from Micropolyspora faeni, Saccharomonospora viridis, Thermoactinomyces candidus, T . sacchari and T . vulgaris were prepared by growing them on dialysate of trypticase soy broth . Sera from rabbits immunized with these antigens were used to study cross-reactivity between thermophilic actinomycetes by antigen-antibody crossed immunoelectrophoresis and by agar gel double diffusion . Mi . faeni and S . viridis showed some degree of cross-reaction, but both failed to show any cross reactivity with Thermoactinomyces species . Antigens from Thermoactinomyces cross-reacted with members within the genus, but no reactivity against S . viridis or M . faeni antisera was detected . Hence, the presence of antibodies to several thermophilic actinomycetes in the sera of patients may be attributed to the exposure of the individual to different thermophilic actinomycetes rather than to the antigenic cross-reactivity between the organisms. Ann Microbiol (Paris), 1979 May-Jun, 130 A(4), 487 - 98 {Amoebae in the water supply: an epidemiological study (author's transl)}; Dive D et al.; Drinking water samples (836) of various origins have been examined for amoebae . For collector water, half of the samples are contamined without influence of the origin . So, occurrence of amoeba in underground waters is discussed . Filtration and disinfection are generally inefficient . Bottles of drinking water are very often polluted during bottling . International mineral waters of french origin are not so damaged during conditioning operation . Bacteria and amoebae populations are quite independant and can't be indicators one for another . Thermophilic amoebae strains are very rare and any pathogenic amoeba has been found during the study. J Protozool, 1979 May, 26(2), 241 - 5 Selection of nonmotile Tetrahymena with Ficoll underlayers; McGwin NF et al.; The mechanisms regulating the development of cilia in Tetrahymena are poorly understood but might be revealed through the study of ciliogenesis mutants . Failure to regenerate cilia after dibucaine deciliation results in continued absence of motility . Therefore, to isolate ciliogenesis mutants efficiently, methods for separating motile and nonmotile cells are essential . We examined the efficacy of Ficoll underlayers for these separations . Ciliates of T . thng type IV) were mixed with Ficoll and added as underlayers to separatory funnels containing growth medium . At 27 C most of the cells remained motile and were found in the top layer; at 37 C, there was a time-dependent increase in the number of nonmotile cells and the number of cells in the Ficoll layer . After 150 min at 37 C, most of the cells became nonmotile and were found in the Ficoll layer . Other studies indicated that at 37 C, the cells remained alive and capable of regenerating cilia when deciliated . Thus, it is clear that the Ficoll underlayer effectively separates the majority of nonmotile cells from the majority of motile cells . Evidently, however, at 37 C wild-type T . thermophila exhibit temperature-sensitive phenotypic variability with regard to motility which should be minimized when selecting for mutations affecting motility and ciliogenesis. J Biochem (Tokyo), 1979 May, 85(5), 1205 - 11 Bacteriophage phiNS11: a lipid-containing phage of acidophilic thermophilic bacteria . IV . Sedimentation coefficient, diffusion coefficient, partial specific volume, and particle weight of the phage; Sakaki Y et al.; The particle weight (molecular weight) of phiNS11 was determined from the sedimentation coefficient, diffusion coefficient, and partial specific volume of the phage . The sedimentation coefficient of the phage (S(0)20, W) is 416 +/- 2.7S . The diffusion coefficient D(0)20, W), which was determined by quasielastic light scattering measurement, is (0.57 +/- 0.03) x 10(-7) cm2/s . The partial specific volume was determined by the mechanical oscillation technique to be 0.747 +/- 0.007 cm3/g . Based on these values, the particle weight of the phage was calculated to be (70.3 +/- 4.3) x 10(6) daltons, which agrees well with the particle weight (69--72 x 10(6) daltons) estimated from the molecular weight of phage DNA and the content of DNA . The Stokes radius of the phage particle was calculated to be 37.7 +/- 2 nm and hydration of the phage was estimated to be 1.18 cm3/g of dry phage . From the particle weight and the chemical composition of the phage, we estimated that one phage particle contains one double-stranded DNA molecule, 16,000 residues of fatty acid, 72 protein I molecules, 920 protein II, 42 protein III, 48 protein IV, 290 protein V molecules, and 3,700 molecules of polyamines. Biochem J, 1979 May 1, 179(2), 407 - 12 Adenosine triphosphate consumption by bacterial arginyl-transfer ribonucleic acid synthetases; Godeau JM et al.; ATP consumption by arginyl-tRNA synthetases from Escherichia coli and Bacillus stearothermophilus has been investigated by the firefly luciferin--luciferase assay . Arginyl-tRNA synthetase from E . coli utilizes ATP only for aminocylation of tRNA with a 1:1 stoicheiometry . In contrast, we have shown an adenosine triphosphatase activity of arginyl-tRNA synthetase from B . stearothermophilus in the absence of tRNAArg . Dowex chromatography revealed the formation of ADP by the thermophile enzyme; under aminoacylation conditions, AMP was also formed in amounts stoicheiometric with arginyl-tRNA formation. Nucleic Acids Res, 1979 Apr, 6(4), 1571 - 81 Role of ribothymidine in the thermal stability of transfer RNA as monitored by proton magnetic resonance; Davanloo P et al.; In order to elucidate the functional role of the modified uridines at position 54 of tRNA, the 270 MHz high-field proton NMR spectra of methionine tRNAs from E . coli, from a mutant thereof, and from T . thermophilus, containing ribothymidine, uridine and 2-thioribothymidine, respectively, have been measured as a function of temperature . A comparison of the NMR melting profiles of the minor nucleosides from these tRNAs shows that the melting temperature of uridine containing tRNA is 6 degrees C lower than that of the wild type tRNA whereas that of the 2-thioribothymidine tRNA is 7 degrees C higher than that of the wild type tRNA . These results, therefore, demonstrate that these modifications serve for stabilization of the tertiary structure of tRNA. J Cell Sci, 1979 Apr, 36, 343 - 53 Dual capacity for nutrient uptake in Tetrahymena . V . Utilization of amino acids and proteins; Orias E et al.; We investigated the relative contributions of phagocytosis and plasma membrane transport to the uptake of amino acids and a protein (egg albumin) in amounts which allow Tetrahymena thermophila to grow and multiply . We used a mutant capable of indefinite growth without food vacuole formation (phagocytosis) and its wild type (phagocytosis-competent) isogenic parental strain . Our results suggest that phagocytosis is not required for free amino acid uptake, most or all of which can be attributed to carrier-mediated transport systems, apparently located on the plasma membrane . In contrast, phagocytosis is required for utilization of the protein . Proteins can supply required amino acids in amounts sufficient for growth only when food vacuoles are formed . We conclude that Tetrahymena thermophila either possesses no endocytic mechanisms at the cell surface other than food vacuole formation or, if it does, these putative mechanisms are not capable of nutritionally meaningful rates of protein uptake. Prax Klin Pneumol, 1979 Apr, 33 Suppl 1, 640 - 3 {Prevention and clinical aspects of pneumoconiosis induced by organic dusts (author's transl)}; Antweiler H; Only those pneumoconioses induced by organic dusts are discussed which are met with in Germany . They comprise: byssinosis which is caused by toxic agents in cotton and flax, farmer's lung, sprayer's fever, exogenous allergic alveolitis due to thermophil actinomyces in the dust of mouldy agricultural meterials or the air of air-conditioning systems, bird fancier's lung which is caused by antigens in animal waste products . Byssinosis and farmer's lung have now been acknowledged as occupational diseases. Mikrobiologiia, 1979 Mar-Apr, 48(2), 222 - 5 {tRNA-methylase study of the extreme thermophile, Thermus flavus}; Shershneva LP et al.; tRNA methylases were studied in the extreme thermophilic culture of Thermus flavus, strain 71 . Like E . coli, the culture contained only those tRNA methylases which catalysed the formation of m1A and m7G . Mg2+, Ca2+ and Na+ ions activated tRNA methylases of Thermus flavus in the series Mg greater than Ca greater than Na while Mn2+ ions inhibited the enzyme . The activity of tRNA methylases was higher in T . flavus than in E . coli, and required less protein and time for exhaustive methylation of tRNA preparations . The overall activity of methylases in T . flavus at 70 degrees C was 5-6 times higher than at 40 degrees C; the elevation of temperature had different effect on various methylases: the activity of m1A methylase increased 13-fold whereas that of m7G methylase only twofold. Biotechnol Bioeng, 1979 Mar, 21(3), 345 - 55 Association of beta-glucosidase with intact cells of Thermoactinomyces; Hagerdal B et al.; The location of the B-glucosidase activity in a whole culture broth of the thermophilic organism Thermoactinomyces has been studied . Little beta-glucosidase activity was found in the culture filtrate, while the culture solids contained the major part of the activity of the whole culture broth . The activity does not appear to be adsorbed to the culture solids; rather there is evidence that it is an intracellular soluble enzyme(s) . The pH and temperature optima for a crude beta-glucosidase preparation were determined to be pH 6.5 and 50--55 degrees C . Enzyme activity studies indicate that the same enzyme(s) accounts for the beta-glucosidase and the cellobiase activities . The validity of using the filter paper activity of culture filtrates from Thermoactinomyces to predict the total saccharification of cellulosic materials to glucose is discussed. Vet Med (Praha), 1979 Mar, 24(3), 185 - 92 {Microbiology of various liver products}; Lukasova J et al.; The proportions of mesophilic, thermophilic and psychrophilic microorganisms were investigated in the total number of microbes and aerobic sporogenic microorganisms in delicatessen liver sausages and liver cheeses . At the same time the content of ammonia and the pH of the products were determined . The largest part of the microflora comprised mesophilic microorganisms, 10 to 25% of which belonged to the sporogenic microflora . The incidence of staphylococci, enterococci and coliform microorganisms was also studied . The total number of microbes contributed to the ammonia content in the products, however, no intercorrelation was statistically proved . In the case of pH no relationship between its value and the number or kind of investigated microorganisms was found . All the indicators were followed in the raw material, in the finished products after heat processing and in products stored at refridgerator and room temperatures for one week. J Am Vet Med Assoc, 1979 Feb 1, 174(3), 277 - 81 The role of allergy in chronic pulmonary disease of horses; Halliwell RE et al.; Twenty-five horses with chronic pulmonary disease were skin tested with allergenic extracts of 24 molds, 4 thermophilic actinomyces, barn dust, hay dust, soya-bean mill dust, and grain mill dust . The results were compared with those obtained on 25 normal horses . Between the 2 groups of horses, there was a highly significant difference in positive skin test results at 30 minutes and 4 hours. Biochem J, 1979 Feb 1, 177(2), 441 - 8 Simple efficient methods for the isolation of malate dehydrogenase from thermophilic and mesophilic bacteria; Wright IP et al.; Malate dehydrogenase from a number of bacteria drawn from several genera and representing the mesophilic, moderately thermophilic and extremely thermophilic classes was isolated by procedures which involve only a small number of steps (in most cases only two), of which the key one is affinity chromatography on 5'-AMP--Sepharose and/or on NAD+--hexane--agarose . Electrophoretic analysis of the native enzymes in polyacrylamide gel and of the denaturated enzymes in sodium dodecyl sulphate/polyacrylamide gel revealed no significant protein impurity in the purified preparations . The yields ranged from about 40% to over 80% . The malate dehydrogenases from the extreme thermophiles and from some of the moderate thermophiles are appreciably less efficient catalytically than their mesophilic homologues. J Biochem (Tokyo), 1979 Feb, 85(2), 503 - 9 Carbodiimide-binding protein of H+-translocating ATPase and inhibition of H+ conduction by dicyclohexylcarbodiimide; Sone N et al.; H+-Translocating ATPase, which catalyzes ATP synthesis in biomembranes, is composed of a head piece (F1) and a membrane moiety (F0) . Using highly-purified F0 from a thermophilic bacterium PS3 (TF0), the following results were obtained . 1 . Inhibition by N,N'-dicyclohexylcarbodiimide (DCCD) of H+ conduction through TF0 followed pseudo-first-order kinetics . The second-order rate constant for inhibitor-enzyme interaction was 5 times 10(3) M(-1)-min(-1) . 2 . H+ conductivity blocked by DCCD was proportional to the amount of DCCD incorporated in the band 8 protein of TF0 . When only one-third of the band 8 protein was labeled with DCCD, TF0 hardly transported any H+ . 3 . By extracting TF0 with chloroform-methanol, the band 8 protein was obtained as a proteolipid . Polyacrylamide gel electrophoresis with dodecyl sulfate and urea showed that the molecular weight was about 6,000 . 4 . The amino acid composition of band 8 protein indicated that this protein contained an extremely high percentage of hydrophobic amino acids (0.29 in polarity) and was devoid of histidine, tryptophan, cysteine, and lysine . Its minimum molecular weight was 6,500 . 5 . The role of band 8 protein (DCCD-binding protein) in H+ conduction through TF0 is discussed on the basis of these results. Biochim Biophys Acta, 1979 Jan 12, 566(1), 62 - 6 Hydrolysis of low molecular weight isomaltosaccharides by a p-nitrophenyl-alpha-D-glucopyranoside-hydrolyzing alpha-glucosidase from a thermophile, Bacillus thermoglucosidius KP 1006; Suzuki Y et al.; A p-nitrophenyl-alpha-D-glucopyranoside-hydrolyzing alpha-glucosidase of a thermophile, Bacillus thermoglucosidius KP 1006, was purified to an electrophoretically-homogeneous state . Its molecular weight was estimated as 60 000 by gel electrophoresis . The molecular activity (ko) and the Km value at 60 degrees C and pH 6.8 for p-nitrophenyl-alpha-D-glucopyranoside were 233 s-1 and 0.24 mM, respectively . The enzyme cleft the non-reducing terminal alpha-1,6-glucosidic bonds of isomaltose, panose, isomaltotriose, isomaltotetraose, and isomaltopentaose . The ko values were 72.4, 194, 208, 233 and 167 s-1, and the Km values were 3.3, 9.5, 11, 13 and 21 mM, respectively . Each isomaltosaccharide was hydrolyzed to glucose by the cleavage of single glucose units from its nonreducing end . The present study suggests that the enzyme is an oligo-1,6-glucosidase (dextrin 6-alpha-glucanohydrolase, EC 3.2.1.10) and an exo-glucosidase. Chromosoma, 1979, 75(3), 293 - 308 An analysis of spindle ultrastructure during prometaphase and metaphase of micronuclear division in Tetrahymena; LaFountain JR Jr et al.; Mitotic micronuclei were isolated from Tetrahymena thermophila in a medium containing hexylene glycol and their ultrastructure was analyzed using thin section techniques . The two stages selected for analysis were early prometaphase and metaphase . A comparison of data from these two stages revealed several differences in nuclear morphology . Metaphase nuclei were longer, they contained more microtubules, and the distribution of microtubules at metaphase was different from that at early prometaphase . Increases in microtubules, which are a unique class of microtubules that can be distinguished from other classes on the basis of their close association to the nuclear membrane . Growth of peripheral sheath microtubules is thought to be significant because it could be the mechanical basis of nuclear elongation . Crossbridges were observed throughout the spindle between all classes of microtubules, but the exact function of these elements remains to be determined. Folia Microbiol (Praha), 1979, 24(5), 396 - 402 Thermomonospora sp . T-SA-125 and its production of a growth promoting antibiotic; Dewedar A et al.; Thermomonospora sp . T-SA-125 is a true thermophilic actinomycete isolated from a soil sample collected from the Saudi Arabian desert . It is characterized by the formation of single spores at the tips of dichotomously branched aerial mycelium and differs from Thermomonospora curvata and T . viridis in certain aspects . It produces a basic water-soluble antibiotic which is active against Gram-positive bacteria, moderately active against Gram-negative bacteria and inactive against fungi . At high concentrations, this antibiotic, stimulated the growth of both Hordeum coleoptile and lettuce hypocotyl. J Embryol Exp Morphol, 1979 Jan, 49, 203 - 27 A mutant of Tetrahymena thermophila with a partial mirror-image duplication of cell surface pattern . II . Nature of genic control; Frankel J et al.; The CU-127 clone of Tetrahymena thermophila, which manifests an unusually high number of ciliary rows plus a second set of abnormal oral structures and of contractile vacuole pores with partial mirror-image reversal of asymmetry (Jerka-Dziadosz & Frankel, 1979), has been subjected to breeding analysis . The progeny ratios obtained in various crosses indicate that the abnormalities of cell-surface asymmetry are brought to expression as a result of the action of a recessive allele at a single gene locus, here named janus . When previously normal cells were made homozygous for the jan allele, the cortical pattern characteristic of the CU-127 clone came rapidly to expression, often without associated change in number of ciliary meridians . Conversely, when cells previously expressing jan re-acquired the wild-type (jan+) allele, they returned to the normal pattern of a single normal oral structure and a single normally located set of contractile vacuole pores while still retaining the high ciliary meridian number characteristic of the original CU-127 clone . The capacity for manifestation of the unique asymmetry pattern depends solely on homozygous expression of the janus allele, whereas the stable number of ciliary meridians in janus clones and the degree of expression of secondary OAs is modulated by other factors, probably at least in part genic . These results, taken together with those of the preceding paper, indicate that the janus allele promotes the propagation and/or expression of a condition of reversed asymmetry in a precisely located cell region, and further indicates that the propagation and expression of this condition are largely independent of the number and asymmetry of ciliary meridians. J Embryol Exp Morphol, 1979 Jan, 49, 167 - 202 A mutant of Tetrahymena thermophila with a partial mirror-image duplication of cell surface pattern . I . Analysis of the phenotype; Jerka-Dziadosz M et al.; Cells of a mutant clone, CU-127, of Tetrahymena thermophila (formerly T . pyriformis, syngen 1) manifest three anatomical abnormalities . First, the stable number of ciliary meridians is 21-25, above the usual number (17-21) in this species . Second, up to 30% of the cells have two oral apparatuses (OAs), one normal and the other abnormal . Third, more than one-half of the cells possess two distinct sets of contractile vacuole pores (CVPs) . In some living cells two contractile vacuoles are seen . These abnormalities have persisted unchanged during more than 500 generations of vegetative propagation, and are similarly expressed in subclones . The normal and abnormal OAs are topographically segregated, with normal OAs developing along the "primary oral axis" and abnormal OAs developing along a "secondary oral axis" that is situated 170 degrees of the cell circumference to the cell's right of the primary oral axis . CVPs always appear within this 170 degree arc and never within the complementary 190 degrees arc to the left of the primary oral axis . A unique feature of the CU-127 clone is the commonly expressed mirror image reversal of the structural pattern of OAs that develop along the secondary oral axis . The primordia of such OAs initially appear (as usual) to the cell's left of a ciliary meridian, but as membranelles develop the frequently come to be oriented in a mirror image of the normal pattern, and an undulating membrane sometimes develops on the wrong (left) side of the oral primordium . When two sets of CVPs are formed, their average positions are roughly equidistant with respect to the two oral axes, with the two sets located 50-60 degrees to the right and left respectively of the primary and secondary oral axis . Such cells are thus bilaterally symmetrical about a plane defined by the central longitudinal axis and the halfway point between the two CVP sets (see Fig . 25) . This plane bisects the cell into a normal and a "reversed" half-cell . However, only oral asymmetry and large-scale CVP positioning are subject to such reversal; all ciliary meridians remain of normal asymmetry and all CVPs are situated on the left side of CVP meridians . The fact that major aspects of large-scale cellular organization can be reversed while the "fine-positioning" associated with the ciliary meridians remains normal indicates that the two aspects of cell organization are distinct. Mikrobiologiia, 1979 Jan-Feb, 48(1), 133 - 6 {Fixation of molecular nitrogen by sulfate-reducing bacteria from petroleum strata}; Nazina TN et al.; The activity of nitrogen fixation by the museum strains of sulfate reducing bacteria isolated from oil deposits was studied using the acetylene technique . The mesophilic sulpfate reducing bacteria belonging to the species Desulfovibrio africanus 2372 and D . baculatus X were found to have a high nitrogenase activity . D desulfuricans subsp . aestuarii 2198 reduced acetylene at a low rate . The thermophilic sulfate reducing cultures of D . thermophilus 7, Desulfotomaculum nigrificans 781 and Dm . nigrificans subsp . salinus 435 produced only small quantities of ethylene . Apparently, nitrogen fixation by sulfate reducing bacteria can be considerable in oil strata whose temperature does not exceed 35--40 degrees C. Clin Allergy, 1979 Jan, 9(1), 43 - 52 Microbiological and serological studies of farmers' lung in Finland; Terho EO et al.; Sera from few Finnish patients with clinical farmers' lung react in precipitin tests with extracts o