Genetic Diversity of Bile Salt Hydrolases Among Human Intestinal Bifidobacteria

Attachment of Brachyspira hyodysenteriae to intestinal epithelial cell lines and its possible mediation by outer membrane proteins (OMPs) of the spirochete were examined. Different B. hyodysenteriae serotypes were shown to adhere to rat and swine intestinal epithelial cells (IEC-18 and IPEC-J2) in vitro but not to the human rectal tumor cell line (HRT-18). Adherence of strain B204 to IPEC-J2 cells was reduced by rOMP-specific antisera in amounts of 29 % (anti-rBhlp29.7), 59 % (anti-rBhlp16), 70 % (anti-rBhmp39h), and 74 % (anti-rBhmp39h), respectively. By use of pooled antisera against Bhlp16 and Bhmp39f inhibition rates of the other serotypes ranged from 53 to 91 %. In a western blot assay OMPs of all serotypes but one were detected by the respective rOMP antisera. Altogether the results indicated that OMPs of B. hyodysenteriae displayed a serotype overlapping antigenicity and mediated adherence of the spirochetes to animal cell cultures.     

PURIFICATION AND CHARACTERIZATION OF A PROTEINASE FROM THE PROBIOTIC Lactobacillus rhamnosus OXY

A proteinase produced by the human gastrointestinal isolate Lactobacillus rhamnosus strain OXY was identified and characterized. The prtR2 gene coding for proteinase activity was detected in the examined strain. The PCR primers used were constructed on the basis of the sequence of the prtR2 proteinase gene from Lactobacillus rhamnosus GG. The enzyme was purified by fast protein liquid chromatography (FPLC) using CM-Sepharose Fast Flow and Sephacryl S-300 columns. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the enzyme had a relatively low molecular mass of 60 kD. Protease activity was observed at a pH range from 6.5 to 7.5 with optimum k _cat/K _m values at pH 7.0 and 40°C. Maximum proteolytic activity (59 U mL^?1) was achieved after 48 hr of cultivation. The activity of the enzyme was inhibited only by irreversible inhibitors specific for serine proteinases (PMSF and 3,4-dichloro-isocumarine), suggesting that the enzyme was a serine proteinase. Proteinase activity was increased by Ca²⁺ and Mg²⁺, and inhibited by Cu²⁺, Zn²⁺, Cd²⁺, and Fe^2+.     

Humans do not perceive conspecifics with a greater exposed sclera as more trustworthy: a preliminary cross-ethnic study of the function of the overexposed human sclera

Understanding the adaptive function of the unique morphology of the human eye, in particular its overexposed white sclera, may have profound implications for the fields of evolutionary behavioural science, and specifically the areas of human interaction and social cognition. Existing hypotheses, such as the cooperative eye hypothesis, have attracted a lot of attention but remain untested. Here, we: (i) analysed variation in the visible sclera size in humans from different ethnic backgrounds and (ii) examined whether intraspecific variation of exposed sclera size is related to trust. We used 596 facial photographs of men and women, assessed for perceived trustworthiness, from four different self-declared racial backgrounds. The size of the exposed sclera was measured as the ratio between the width of the exposed eyeball and the diameter of the iris (sclera size index, SSI). The SSI did not differ in the four examined races and was sexually monomorphic except for Whites, where males had a larger SSI than females. In general, the association between the SSI and trustworthiness was statistically insignificant. An inverted U-shaped link was found only in White women, yet the strength of the effect of interaction between sex and race was very small. Our results did not provide evidence for the link between exposed sclera size and trustworthiness. We conclude that further investigation is necessary in order to properly assess the hypotheses relating to the socially relevant functions of overexposed sclera.     

Occurrence of UDPG: Sterol glucosyltransferase activity in some lower plants

A study was made of the sterol glucosylating ability of cell-free homogenates obtained from 16 species of photosynthesizing and nonphotosynthesizing lower plants (2 species of Chlorophyceae, 2 species of Cyanophyceae, 1 species of Phycomycetes, 3 species of Ascomycetes, 3 species of Basidomycetes, 1 species of Myxomycetes, 3 species of Musci and 1 species of Sphenopsida). Except for the blue-green and green algae, all the remaining species showed distinct in vitro synthesis of steryl monoglucosides from UDPG and cholesterol or sitosterol. Preliminary studies on the specificity of the relevant enzymes pointed to a correlation between the sterol composition of the plant and the specificity of its glucosylating enzyme. The enzyme from Ascomycetes and Basidomycetes utilized ergosterol better than cholesterol or sitosterol. Enzymic preparations from mosses utilized sitosterol the best.     

Genotoxicity of alpha-asarone analogues

The role of cholesterol in the formation of atherosclerotic lesions during hypercholesterolemia has been confirmed. alpha-Asarone is a substance of a potent hypolipidemic activity which is isolated from plants. We previously described the synthesis of several alpha-asarone analogues exhibiting hypolipidemic and antiplatelet activity. Genotoxic activity of four selected alpha-asarone analogues was theoretically evaluated based on quantum-mechanical method for calculation of enthalpy of carbocations formation (DeltaH(R)) according to the Testa's method. In the present paper, we evaluated the mutagenic and genotoxic activity of alpha-asarone isomers 2-5 based on the reference Ames test and micronucleus test. Results obtained in the study show that tested isomers were non-mutagenic, however, they exhibited growing cytotoxic activity. Relationship between the heat of formation of their putative metabolic intermediates and mutagenic/genotoxic activity was not confirmed.     

Egg formation and the early embryonic development of Aspidogaster limacoides Diesing, 1835 (Aspidogastrea: Aspidogastridae), with comments on their phylogenetic significance

Ultrastructural aspects of the early embryonic development of the aspidogastrean Aspidogaster limacoides are described and their phylogenetic implications discussed. Whereas the proximal regions of the uterine lumen usually contain unembryonated eggs or eggs with early embryos, the posterior or distal regions of the uterus are filled with eggs containing a fully-developed cotylocidium. The eggs of A. limacoides can be classified as polylecithal due to the presence of numerous vitellocytes which accompany each fertilized oocyte or ovum during egg formation. The results of the study are described in details under six headings: (1) general characteristics of the intrauterine eggs; (2) eggshell and operculum formation; (3) unembryonated eggs; (4) zygote formation and early cleavage divisions; (5) embryonic envelope formation; and (6) early degeneration or apoptosis of some blastomeres. The late differentiation of the operculum, possible functions of GER-bodies, and the early degeneration of vitellocytes and some blastomeres in this species are compared, drawn and discussed with corresponding observations reported for other parasitic Platyhelminthes. The most important differences are apparent in the number of egg envelopes and their mode of formation in A. limacoides compared with previous reports for both digeneans and cestodes. The results of the present TEM study indicate that the three macromeres, resulting from two cleavage divisions, take part in the formation of a single embryonic outer envelope in A. limacoides, and that this takes place at a very early stage of embryogenesis. Their fusion results in the formation of a single continuous cytoplasmic layer surrounding the early embryo, which is composed of only a small number of undifferentiated blastomeres. The early separation of the macromeres may indicate an equal cleavage pattern. These results suggest that the systematic position of the Aspidogastrea among the Platyhelminthes still remains somewhat equivocal, and indicate the need for more studies on the embryonic development, larval morphogenesis and molecular phylogeny for the elucidation of the relationships between this enigmatic group and related taxa.     

Ampicillin resistance in Listeria monocytogenes acquired as a result of transposon mutagenesis

We have used plasmid pLTV3, which carries transposon Tn917, to obtain a series of mutants of Listeria monocytogenes EGD showing varied degrees of resistance to ampicillin and other beta-lactam antibiotics, including imipenem. In this paper we focus on the characteristics of two strains in which decreased susceptibility to ampicillin is accompanied by changes in the structure of the cell wall murein and cell-wall related changes of phenotype.     

Carbonic anhydrase inhibitors. Selective inhibition of human tumor-associated isozymes IX and XII and cytosolic isozymes I and II with some substituted-2-mercapto-benzenesulfonamides

A series of 2-mercapto-substituted-benzenesulfonamides has been prepared by a unique two-step procedure starting from the corresponding 2-chloro-substituted benzenesulfonamides. Compounds bearing an unsubstituted mercapto group and the corresponding S-benzoyl derivatives were investigated as inhibitors of four isoforms of the zinc enzyme carbonic anhydrase (CA, EC 4.2.1.1), i.e., the cytosolic, ubiquitous isozymes CA I and II, as well as the transmembrane, tumor associated isozymes CA IX and XII. These derivatives were medium potency hCA I inhibitors (K(I)s in the range of 1.5-5.7 microM), two derivatives were strong hCA II inhibitors (K(I)s in the range of 15-16 nM), whereas the others showed weak activity. These compounds inhibited hCA IX with inhibition constants in the range 160-1950 nM and hCA XII with inhibition constants in the range 1.2-413 nM. Some of these derivatives showed a certain degree of selectivity for inhibition of the tumor-associated over the cytosolic isoforms, being thus interesting leads for the development of potentially novel applications in the management of hypoxic tumors which overexpress CA IX and XII.