Revisione delle specie: Actinomyces albus,A. chromogenus,A. odorifer,A. thermophylus,A. viridis,A. viridochromogenes,A. hominis,A. innominatus

Riassunto Lo studio comprende una revisione critico-sperimentale della specie Actinomyces albus, della quale vengono considerati come sinonimi circa 30 nomi speeifici, fra i quale A. chromogenus, A. odorifer, A. thermophylus p.p.; della specie é data una diagnosi ed una particolareggiata descrizione.Sono inoltre studiate le specie A. viridis, (= A. viridochromogenus) e A. innominatus, n. nomen. Quest'ultima é preceduta da una breve discussione sulla specie A. homini.

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Summary Twenty-six strains of Actinomyces albus are studied redescribed from morphological, cultural and biochemical standpoints. Many biological activities of A. chromogenus, A. odorifer and A. thermophilus are in common with other species of the same genus, so that they may be considered for sub-specific, (not specific) differentiation. A discussion on A. farcinicus, A. albidoflavus and A. aureus has been originated from mislabeling as A. albus; the group including the two last named species (flavus group) must be revised. A few strains classified A. farcinicus are in no doubt true A. albus, but this real specific entity remains to be revised from Nocard's strain. A. viridis, for the first time described by Lombardo-Pellegrino, has been redescribed three times as a new species under the same binomial, and the fourth as A. viridochromogenes. A. hominis Bostroem is an uncorrect determination for the species originally described by Waksmann and Curtiss, and it is renamed A. innominatus, the binomial A. (Streptothrix) hominis Auct. being a nomen ambiguum. In conclusion, 30 bionmial are appended in sinonimy to A. albus, including Cladothrix dichotoma Macé (1888) non Cohn, G. invulnerabilis Acosta et G. Rossi, C. odorifera Rullm. Actinomyces chromogenus Gasp., A. thermophilus Auct., p.p., A. (Streptothrix) Sanninii (Cif.) Westerd., A. Almquisti Duché, A. Gougeroti Duché, and so on.

     

Identification of gibberellins A17, A25, A45, abscisic acid,phaseic acid,and dihydrophaseic acid in seeds of Pyrus communis

Gibberellin A₁₇, abscisic acid, and 4′-dihydrophaseic acid were identified by GC-MS of derivatized extracts from both immature and mature seeds of pear. Immature seeds also contained phaseic acid, gibberellins A₂₅ and A₄₅, and two presumed mono-hydroxylated derivatives of GA₄₅, one of which was tentatively identified as 3β-hydroxy-GA₄₅. Several presumed metabolites of abscisic acid were detected in both mature and immature seeds.     

A single,phosphate-repressible deoxyribonuclease,DNase A,secreted inAspergillus nidulans

High levels of nuclease activities were identified in filtrates ofAspergillus cultures after growth in low- but not in high-phosphate media. Deoxyribonuclease activities, characterized extensively by column chromatography, showed a coincident single peak for ss- and ds-DNase which was distinct from the peak for RNase. Both ss-DNase and ds-DNase are endonucleolytic and showed the highest activity in the presence of Ca²⁺ and Mn²⁺ (atpH 8.0). They also showed identical heat sensitivities suggesting that a single, phosphate-repressible DNase was secreted. This enzyme, therefore, corresponds to the well-characterized extracellular DNase A ofNeurospora. However, theAspergillus DNase A did not cross-react with antisera to secretedNeurospora nucleases and showed different chromatographic properties, and active peptides of different sizes were visualized on DNA activity gels. The increasing derepression ofAspergillus DNase A by decreasing phosphate levels was similar to that of secreted alkaline phosphatase and these increases were both abolished by the regulatory mutantpalcA. This investigation was supported by Grant A2564 from the Natural Science and Engineering Research Council of Canada.     

Efficacy of Antibiotics A204 Sodium, A28695A, and A204np against Coccidia of Rabbits

SYNOPSIS. Three polycyclic, polyether, monocarboxylic acid antibiotics produced by and extracted from a strain of Streptomyces albus and designated A204 sodium, A28695A, or A204np, were specifically tested for activity against hepatic and incidentally against intestinal coccidia of rabbits. All were effective in preventing infections and acceptable to young rabbits when prepared in pelleted feed. Weight gain, however, was not as good in inoculated, medicated rabbits as in noninoculated, nonmedicated controls.
No gross pathologic changes were detected except in the livers of inoculated, nonmedicated controls which were severely infected and greatly enlarged. Livers of medicated rabbits were uninfected and normal in appearance and size.     

Paper chromatographic separation of alpha-ecdysone, ecdysterone, inokosterone, makisterone A and ponasterone A

Three two-phase chromatographic solvent systems, utilizing benzene (b), toluene (T) and alcohol/water mixtures (2-propanol = iP, 2-butanol = 2B, percent alcohol indicated), BiP45, TiP50 and T2B50 will separate ecdysterone, inokosterone, α-ecdysone, makisterone A and ponasterone A Cl). The reproducibility of the Rf's and resolution of these systems and two others tested were satisfactory with the difficult to separate ecdysterone-inokosterone pair. ³H-Ecdysterone label was recovered efficiently (86–94%) from paper chromatograms by ethanol elution.     

A New Metabolite,Parasiticolide A,from Aspergillus parasiticus

Two β-glucosidases, G1 and G2, were purified from the culture supernatant of Penicillium herquei Banier and Sartory. Both the purified enzymes were homogeneous on polyacrylamide disc gel electrophoresis. The molecular weights of G1 and G2 were estimated to be 125,000 and 122,000, respectively, by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. G1 and G2 contained 12.7% and 16.1% carbohydrate as glucose, and had isoelectric points of 5.02 and 5.24, respectively. Both enzymes had optimum pHs of 4.0~4.5 and optimum temperatures at 60°C, but pH - and thermo-stabilities of G1 were higher than those of G2. Both enzymes were active not only on p-nitrophenyl β-d-glucopyranoside, salicin, and the p-glucobioses tested but also on laminarin. CM-Cellulose was a very poor substrate for both enzymes. The activities of G1 toward the substrates except for laminarin and CM-cellulose were apparently higher than those of G2. Both enzymes acted on cellobiose to produce a transfer product.     

Patterns of intraspecific variation inAntennaria alborosea,A. corymbosa,A. marginata,A. microphylla,A. parvifolia,andA. umbrinella (Asteraceae:Inuleae)

Patterns of intraspecific variation were examined inAntennaria alborosea A. E. Porsild,A. corymbosa E. Nels,A. marginata Greene,A. microphylla Rydb.,A. parvifolia Nutt., andA. umbrinella Rydb. AlthoughA. alborosea was initially considered arctic in distribution, it became apparent that a southern montane element also exists. Our results suggest that morphological differences between arctic and southern montane specimens represent clinal variation. The additional morphological data for specimens that occur more than 1,500 km south of the species' range as it was initially described result in a better understanding of this once presumed arctic taxon. Morphological variation in the dioecious speciesA. corymbosa, A. marginata, A. microphylla, A. parvifolia, andA. umbrinella was greater between the genders than was geographic variation within each gender. These results demonstrate that both pistillate and staminate specimens must be examined in dioecious species ofAntennaria if morphological variation in the respective species is to be fully understood. Character size or number of broadly distributed species (A. microphylla andA. parvifolia) generally decreased with increasing longitude, whereas characters of species with more restricted distributions (A. alborosea, A. corymbosa, andA. marginata) generally increased in size or number with increasing latitude or longitude.Antennaria umbrinella was an exception in this respect.     

Evaluation of taxonomic validity of four species of Acanthamoeba: A. divionensis, A. paradivionensis, A. mauritaniensis, and A. rhysodes, inferred from molecular analyses

The taxonomy of Acanthamoeba spp., an amphizoic amoeba which causes granulomatous amoebic encephalitis and chronic amoebic keratitis, has been revised many times. The taxonomic validity of some species has yet to be assessed. In this paper, we analyzed the morphological characteristics, nuclear 18s rDNA and mitochondrial 16s rDNA sequences and the Mt DNA RFLP of the type strains of four Acanthamoeba species, which had been previously designated as A. divionensis, A. parasidionensis, A. mauritaniensis, and A. rhysodes. The four isolates revealed characteristic group II morphology. They exhibited 18S rDNA sequence differences of 0.2-1.1% with each other, but more than 2% difference from the other compared reference strains. Four isolates formed a different clade from that of A. castellanii Castellani and the other strains in morphological group II on the phylogenetic tree. In light of these results, A. paradivionensis, A. divionensis, and A. mauritaniensis should be regarded as synonyms for A. rhysodes.     

A tyrosinase inhibitor, Daedalin A, from mycelial culture of Daedalea dickinsii

A chromene-type compound, daedalin A (1), was isolated from mycelial culture broth of Daedalea dickinsii. Based on spectroscopic data, the structure of 1 was found to be (2R)-6-hydroxy-2-hydroxymethyl-2-methyl-2H-chromene. Daedalin A (1) strongly inhibited the activity of tyrosinase (IC(50): 194 muM). In addition, 1 also showed 1,1-diphenyl-2-picrylhydrazyl scavenging activity (IC(50): 16.9 microM) and superoxide anion scavenging activity (IC(50): 28.5 microM).     

Interaction of growth retardants,daylength, and gibberellins A19, A20, and A1 on shoot elongation in birch and alder

Gibberellins A₁₉, A₂₀, and A₁ were applied to seedlings of birch (Betula pubescens Ehrh.) and alder (Alnus glutinosa (L.) Gaertn.) in order to test their ability to counteract growth inhibition induced by growth retardants (ancymidol and BX-112) or short day (SD, 12 h) photoperiod. Ancymidol inhibits early and BX-112 inhibits late steps in gibberellin biosynthesis. BX-112 inhibited stem elongation in both species while ancymidol, applied as a soil drench, was effective in alder only. Growth retardants affected stem elongation mainly by inhibiting elongation of internodes. All three gibberellins were equally active when applied to seedlings treated with ancymidol; however, only GA₁ was able to counteract the growth inhibition induced by BX-112. SD-induced cessation of elongation growth in birch was counteracted by GA₁, and to some degree, by GA₂₀, while GA₁₉ was inactive. SD treatment did not induce cessation of apical growth in alder. These results are consistent with the hypothesis that of gibberellins belonging to the early C-13 hydroxylation pathway, GA₁ is the only active gibberellin for stem elongation.     

Four alkaloids, lucidine B, oxolucidine A, lucidine A, and lucidulinone from Lycopodium lucidulum

The structures of four alkaloids extracted from Lycopodium lucidulum (Lycopodiaceae) were established by X-ray and 2D NMR spectroscopic analyses. The dihydro-derivative of oxolucidine A, which was obtained by NaBH4 reduction of oxolucidine A, was treated with p-bromobenzoyl chloride to afford crystals, whose X-ray crystallographic analysis established the stereostructure, including the absolute configuration. The 2D NMR spectra of tetrahydrodeoxylucidine B were fully analyzed to establish the full structure of lucidine B, and the hitherto unknown stereochemistry at the C-14 position was established as beta-H. The structure of a new alkaloid, lucidulinone, was determined by spectroscopic analysis to be luciduline lactam.     

A Cytopathogenic, Apoptosis-Inducing Variant of Hepatitis A Virus

A cytopathogenic variant of hepatitis A virus (HAV_cyt/HB1.1) was isolated from persistently infected BS-C-1 cells by serial passages in FRhK-4 cells. This virus shows a rapid replication pattern and high final titers are obtained, which are main characteristics of cytopathogenic HAVs. Sequencing of the nontranslated regions and the coding regions for 2ABC and 3AB revealed that mutations are distributed all over these regions and that certain mutated sites correspond to those in other cytopathogenic HAV variants. Investigating the mechanisms causing the cytopathic effect in FRhK-4 cells infected with this variant, we found that an apoptotic reaction takes place.     

六种鲟鱼消化酶活性的比较研究

测定了两个生长阶段 6种鲟鱼幼鱼胃、肠道和肝脏中蛋白酶、脂肪酶、淀粉酶活性。幼鲟消化酶活性在两个生长阶段变化不明显。 6种鲟鱼不同消化器官蛋白酶活性以肠道为最高 ,肝脏为最低 ,肝脏中的蛋白酶活性明显低于胃、肠道 (P <0 0 1)。不同消化器官脂肪酶活性 ,以肠道为最高 ,且肠道脂肪酶活性显著高于胃、肝脏 (P <0 0 1) ,胃中的脂肪酶活性与肝脏中的脂肪酶活性差异不明显 (P >0 0 5 )。不同消化器官淀粉酶活性 ,以肠道为最高 ,且明显高于胃、肝脏 (P <0 0 1)。幼鲟在第一阶段 ,肝脏中没有淀粉酶活性 ,其活性出现在第二阶段 ,且在此生长阶段 ,肝脏中的淀粉酶活性达到胃中的水平 (P >0 0 5 )。对 6种鲟鱼而言 ,除个别存在较大差异外 ,3种消化酶活性大体上都没有明显差异     

Molecular analysis of the serologically defined HLA-Aw19 antigens. A genetically distinct family of HLA-A antigens comprising A29, A31, A32, and Aw33, but probably not A30

The HLA-Aw19 complex consists of a number of serologically cross-reactive Ag (i.e., A29, A30, A31, A32, and Aw33) which exhibit an epitope shared by HLA-B and -C proteins. To investigate the structural basis for these serologic cross-reactivities, we have cloned and determined the nucleotide sequences for A30, A31, and Aw33, and compared the predicted amino acid sequences with those already available for A29, A32, and other class I allelic products. All alleles of the Aw19 group contained A-locus-specific sequences, exhibiting "A-ness." The structural similarities between Aw19 polypeptides were found in the alpha 1 and alpha 2 domains, where shared amino acid residues were identified that correlated with observed serological reactivity patterns. Seven Aw19-specific nucleotides were found. Two of these were silent substitutions, but the remaining five resulted in Aw19-specific amino acid residues. Each of the HLA-A alleles can be classified into one of the five serologically cross-reacting groups. In the Aw19 group, the alleles A29, A31, A32, and Aw33 are closely related serologically as well as genetically whereas A30 probably belongs to the A1/A3/A11 group. The similarity between A30 and the other Aw19 alleles may have resulted from two independent gene conversions affecting exons 2 and 3. Additional mutations or gene conversion-like events in A30 were also noted. It is postulated that gene conversions have played a significant role in the divergence of the Aw19 alleles. However, each serologically cross-reactive Aw19 allotype appears to have arisen directly from a common ancestral allele. A30 was the only exception, and this allele may represent an unusual allotype, which is subject to a high rate of genetic changes, as is seen in the H-2Kb gene of the mouse.     

MAGE-A1,A2,A3,A4在膀胱移行细胞癌组织及细胞株中基因表达的研究

目的研究膀胱移行细胞癌(TCC)中黑色素瘤抗原(MAGE)基因表达。方法逆转录聚合酶链反应(RT-PCR)技术检测20例膀胱TCC患者癌组织和3株膀胱TCC细胞株T24、EJ、BIU87中MAGE-A1、A2、A3、A4基因mRNA表达。结果20例膀胱TCC癌组织中19例(95%)至少表达一种MAGE-A基因,12例MAGE-A1阳性(60%),16例MAGE-A2阳性(80%),11例MAGE-A3阳性(55%),18例MAGE-A4阳性(90%),MAGE-A1-4均阳性8例(40%)。膀胱TCC细胞株T24中MAGE-A1-4基因均表达,EJ中MAGE-A3、A4基因表达,BIU87中MAGE-A2、A3、A4基因表达。结论MAGE基因在膀胱TCC中有较高表达,可望成为膀胱TCC免疫治疗的靶基因。     

Substrate specificities of cathepsin A,L and A,S from pig kidney

The substrate specificities of two different molecular sizes of cathepsin A, A,L (large form) and A,S (small form), for synthetic substrates were examined kinetically. Both enzymes showed a similar broad substrate specificity against various acyl dipeptides, amino acid esters, and amino acid amides. Z-Phe-Ala and Ac-Phe-OEt were good substrates. Peptides containing hydrophobic amino acids were hydrolyzed rapidly. The presence of hydrophobic amino acid residues, not only at the C-terminal position but also at the second position and probably the third position from the C-terminal, resulted in an increase in the rate of hydrolysis. Peptides containing glycine and proline were hydrolyzed slowly. Inhibition studies with Z-D-Phe-D-Ala and Z-Phe suggested that the peptidase and esterase activities of the enzymes are both catalyzed by the same site of the enzyme molecule, but it remains to be elucidated whether or not the binding sites for peptides and esters are the same.