Speciation of Campylobacter coli, C. jejuni, C. helveticus, C. lari, C. sputorum, and C. upsaliensis by matrix-assisted laser desorption ionization-time of flight mass spectrometry

Multiple strains of Campylobacter coli, C. jejuni, C. helveticus, C. lari, C. sputorum, and C. upsaliensis isolated from animal, clinical, or food samples have been analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Whole bacterial cells were harvested from colonies or confluent growth on agar and transferred directly into solvent and then to a spot of dried 3-methoxy-4-hydroxycinnamic acid (matrix). Multiple ions in the 5,000- to 15,000-Da mass range were evident in spectra for each strain; one or two ions in the 9,500- to 11,000-Da range were consistently high intensity. "Species-identifying" biomarker ions (SIBIs) were evident from analyses of multiple reference strains for each of the six species, including the genome strains C. jejuni NCTC 11168 and C. jejuni RM1221. Strains grown on nine different combinations of media and atmospheres yielded SIBI masses within +/-5 Da with external instrument calibration. The highest-intensity C. jejuni SIBIs were cytosolic proteins, including GroES, HU/HCj, and RplL. Multiple intraspecies SIBIs, corresponding probably to nonsynonymous nucleotide polymorphisms, also provided some intraspecies strain differentiation. MALDI-TOF MS analysis of 75 additional Campylobacter strains isolated from humans, poultry, swine, dogs, and cats revealed (i) associations of SIBI type with source, (ii) strains previously speciated incorrectly, and (iii) "strains" composed of more than one species. MALDI-TOF MS provides an accurate, sensitive, and rapid method for identification of multiple Campylobacter species relevant to public health and food safety.     

Speciation of Campylobacter coli, C. jejuni, C. helveticus, C. lari, C. sputorum, and C. upsaliensis by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

Multiple strains of Campylobacter coli, C. jejuni, C. helveticus, C. lari, C. sputorum, and C. upsaliensis isolated from animal, clinical, or food samples have been analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Whole bacterial cells were harvested from colonies or confluent growth on agar and transferred directly into solvent and then to a spot of dried 3-methoxy-4-hydroxycinnamic acid (matrix). Multiple ions in the 5,000- to 15,000-Da mass range were evident in spectra for each strain; one or two ions in the 9,500- to 11,000-Da range were consistently high intensity. “Species-identifying” biomarker ions (SIBIs) were evident from analyses of multiple reference strains for each of the six species, including the genome strains C. jejuni NCTC 11168 and C. jejuni RM1221. Strains grown on nine different combinations of media and atmospheres yielded SIBI masses within ±5 Da with external instrument calibration. The highest-intensity C. jejuni SIBIs were cytosolic proteins, including GroES, HU/HCj, and RplL. Multiple intraspecies SIBIs, corresponding probably to nonsynonymous nucleotide polymorphisms, also provided some intraspecies strain differentiation. MALDI-TOF MS analysis of 75 additional Campylobacter strains isolated from humans, poultry, swine, dogs, and cats revealed (i) associations of SIBI type with source, (ii) strains previously speciated incorrectly, and (iii) “strains” composed of more than one species. MALDI-TOF MS provides an accurate, sensitive, and rapid method for identification of multiple Campylobacter species relevant to public health and food safety.     

Stabilities of consecutive A.C, C.C, G.G, U.C, and U.U mismatches in RNA internal loops: Evidence for stable hydrogen-bonded U.U and C.C.+ pairs.

The stability and structure of RNA duplexes with consecutive A.C, C.A, C.C, G.G, U.C, C.U, and U.U mismatches were studied by UV melting, CD, and NMR. The results are compared to previous results for GA and AA internal loops [SantaLucia, J., Kierzek, R., & Turner, D. H. (1990) Biochemistry 29, 8813-8819; Peritz, A., Kierzek, R., & Turner, D.H. (1991) Biochemistry 30, 6428-6436)]. The observed order for stability increments of internal loop formation at pH 7 is AG = GA approximately UU greater than GG greater than or equal to CA greater than or equal to AA = CU = UC greater than or equal to CC greater than or equal to AC. The results suggest two classes for internal loops with consecutive mismatches: (1) loops that stabilize duplexes and have strong hydrogen bonding and (2) loops that destabilize duplexes and may not have strong hydrogen bonding. Surprisingly, rCGCUUGCG forms a very stable duplex at pH 7 in 1 M NaCl with a TM of 44.8 degrees C at 1 x 10(-4) M and a delta G degrees 37 of -7.2 kcal/mol. NOE studies of the imino protons indicate hydrogen bonding within the U.U mismatches in a wobble-type structure. Resonances corresponding to the hydrogen-bonded uridines are located at 11.3 and 10.4 ppm. At neutral pH, rCGCCCGCG is one of the least stable duplexes with a TM of 33.2 degrees C and delta G degrees 37 of -5.1 kcal/mol. Upon lowering the pH to 5.5, however, the TM increases by 12 degrees C, and delta G degrees 37 becomes more favorable by 2.5 kcal/mol. The pH dependence of rCGCCCGCG may be due to protonation of the internal loop C's, since no changes in thermodynamic parameters are observed for rCGCUUGCG between pH 7 and 5.5. Furthermore, two broad imino proton resonances are observed at 10.85 and 10.05 ppm for rCGCCCGCG at pH 5.3, but not at pH 6.5. This is also consistent with C.C+ base pairs forming at pH 5.5. rCGCCAGCG and rGGCACGCC have a small pH dependence, with TM increases of 5 and 3 degrees C, respectively, upon lowering the pH from 7 to 5.5. rCGCCUGCG and rCGCUCGCG also show little pH dependence, with TM increases of 0.8 and 1.4 degrees C, respectively, upon lowering the pH to 5.5.(ABSTRACT TRUNCATED AT 400 WORDS)     

Analysis of morphological characters of Chara baltica, C. hispida, C. horrida, and C. rudis from Europe

Morphological characters of Chara baltica, C. hispida, C. horrida, and C. rudis from different localities in Central Europe and Scandinavia were analyzed. The plants were preliminarily classified using specific qualitative features into four species: C. baltica, C. hispida, C. horrida, and C. rudis. Of the 14 characters, all were used in principal-component and cluster analyses, and 12 were applied in a discriminant analysis. In the discriminant analysis, the most useful characters for distinguishing particular species were selected. The methods (canonical discriminant and canonical analyses) used herein offered good differentiation between brackish water species C. horrida and C. baltica and fresh water species C. hispida and C. rudis. The analysis showed that the two morphologically very similar species C. hispida and C. rudis are only partially distinguished.     

Étude Électrophorétique Et Immunochimique Comparée Des Antigènes De Quelques Levures Du GenreCandida (C. Albicans,C. Stellatoidea,C. Tropicalis,C. Zeylanoides,C. Krusei,C. Pseudotropicalis,C. Macedoniensis)

Résumé L'électrophorèse en gélose pratiquée sur les antigènes des espèces deCandida étudiées, met en évidence 6 fractions. La séparation bien que sommaire a permis l'appréciation de la nature chimique de ces antigènes. La zone de faible mobilité anodique constitue le composant majeur de nature essentiellement protéique, tandis que la fraction lente est intensément révélée par la coloration à l'acide périodique-réactif de Schiff. L'électrophorèse en gel d'amidon, grâce au grand pouvoir de résolution du support a fourni une meilleure séparation des protéines. Intéressants mais insuffisants, les résultats acquis par ces deux types d'électrophorèse de zone sont très largement complétés par ceux de l'immunoélectrophorèse. Cette dernière méthode d'une haute spécificité encore qu'elle reste tributaire de la qualité des immunsérums employés a conduit à un dénombrement précis des fractions antigéniques desCandida puis à la mise en évidence des antigènes communs et spécifiques des espèces étudiées. Les résultats obtenus confirment ceux que nous avons obtenus précédemment (Biguet & coll., 1959a, 1959b, 1960) quant aux affinités réciproques de certaines espèces. Reposant sur des bases physico-chimiques très différentes, ils rejoignent et confirment les travaux extrêment importants de l'école japonaise deTsuchiya et apportent un nouvel argument immunologique à l'hétérogénéité du genreCandida. Sous l'angle taxonomique au moins, il semble que notre méthode présente sur celle des auteurs japonais l'avantage de révéler un nombre beaucoup plus considérable de fractions, partant de permettre en particulier la mise en évidence de divergences de structure qui échapperaient par l'utilisation de leur technique (par exemple, celle qui séparentC. pseudotropicalis deC. macedoniensis).

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Summary Agar electrophoresis carried out on antigens ofCandida species studied yielded six fractions. The separation, though summary, permitted the evaluation of the chemical nature of the antigens. The zone of the weak anionic motility constitutes the major component, essentially proteinic in nature, while the slow fraction is intensely revealed by the PAS stain. Starch gel electrophoresis, due to its great power of resolution, yielded better separation of proteins. The results obtained by these two types of zone electrophoresis, interesting but not sufficient, are a great deal complemented by the results of immuno-electrophoresis. The latter method of high specificity, even though it depends on the quality of the immune serum used, led to a precise enumeration of the antigenic fractions ofCandida, furthermore to the evidence of common and specific antigens of the species studied. The results obtained confirmed those previously obtained (Biguet et coll., 1959a, 1959b, 1960) concerning the reciprocal affinities of certain species. Resting on very different physico-chemical bases, they confirm the extremely important work of the school ofTsuchiya and bring a new immunologic argument as to the heterogeneity of the genusCandida. At least under taxonomic point of view, it seems that our method shows an advantage compared with that of the Japanese authors in revealing a much greater number of fractions, consequently in permitting the proof of divergence of structure which would escape by using their technique (e.g. those which separateC. pseudotropicalis fromC. macedoniensis).

     

Cronisia and its two species,C. fimbriata and C. weddellii

The systematic position of Riccia fimbriata Nees, a species formerly known only in sterile condition and tentatively placed in the genus Exormotheca, is reexamined in the light of the recently discovered reproductive structures. The species proved to be a member of the genus Cronisia, identical to Cronisia mexicana Hicks, described in 1993. A key is provided to the two known species of Cronisia, C. fimbriata and C. weddellii, and they are described and illustrated.     

Photosynthesis,Transpiration, and Water Use Efficiency of Caragana microphylla,C. intermedia,and C. korshinskii

In the order C. microphylla — C. intermedia — C. korshinskii, compensation irradiance, saturation irradiance, and optimum temperature for photosynthesis increased, net photosynthetic rate (P _N) at low irradiance and low temperature decreased, optimum air humidity decreased, and P _N at low air humidity increased. Daily cumulative value of P _N increased while daily cumulative value of transpiration (E) decreased, and hence water use efficiency (WUE =P _N/E) increased. Diurnal course of P _N of C. microphylla was a double-peak curve, but the second peak in the curves of C. intermedia and C. korshinskii was not visible. These physiological characteristics are biological basis for the geographical distribution of these three Caragana species, and are in relation to water conditions of their habitats and distinctiveness in leaf hair of plant.     

Dreaming,fenfluramine, and vitamin C.

The effect of increasing doses of fenfluramine on dream patterns was studied in 20 patients receiving a reducing diet with or without a controlled dietary intake of vitamin C daily. The dream pattern was unchanged in six patients and dreams disappeared in another who normally dreamed often. In 13 patients dreams increased in frequency and intensity, and in five the dreams assumed frightening proportions. There was a significant straight-line relation between response and the size of the dose. When placebo tablets were given to four patients their dreams disappeared or assumed their pretreatment normal pattern. Absence of vitamin C from the diet did not significantly affect the dream pattern. That fenfluramine has dose-related cerebral effects should be remembered in patients with a history of mental illness.