Synthesis, surface and antimicrobial properties of some quaternary ammonium homochiral camphor sulfonamides.

A group of homochiral quaternary ammonium sulfonamides bearing hydrophobic camphor derived moieties were synthesized and characterized. The described synthetic procedure is quick and efficient. The novel quaternary ammonium bromides were tested as antimicrobial and antifungal agents. They exhibited strong antimicrobial and also antifungal activity, especially N-{2-[((1S, 4R)-7,7-dimethyl-2-oxobicyclo[2.2.1]heptan-1-yl)methylsulfonamido] ethyl}-N,N-dimethyltetradecan-1-aminium bromide 1c. The surface properties of prepared compounds were evaluated by surface tension measurements and critical micelle concentration (CMC) with surface tension at CMC (γCMC) was calculated.

Immunodetection and N-terminal sequencing of DNA replication proteins of bacteriophage BFK20 - lytic phage of Brevibacterium flavum.

Phages are excellent models for studying the mechanism of DNA replication in prokaryotes. Identification of phage proteins involved in phage DNA replication is the first prerequisite for elucidation of the phage replication module. We focused on replication proteins gp41 (a putative helicase from SF2 superfamily), gp43 (a RepA-like protein), and gp44 (a putative DNA polymerase A) of phage BFK20 grown in Brevibacterium flavum. To identify them in the phage-host system, we prepared antibodies to these proteins which were cloned and expressed in Escherichia coli as his-tagged recombinant proteins. After purification to homogeneity the recombinant proteins served for raising specific polyclonal antibodies in mice. Using these antibodies in Western blot analysis the phage proteins gp41, gp43 and gp44 were detected during the phage growth cycle. The proteins gp41 and gp43, prepared from cell lysate by ammonium sulphate precipitation, were N-terminally sequenced and found to contain the sequences N-SVKPRELR-C and N-MLGSTML-C, respectively. This means that gp41 starts with serine but not with common methionine. We consider these findings an initial but important step towards more thorough characterization of replication proteins of phage BFK20.

Identification and biological activity of potential probiotic bacterium isolated from the stomach mucus of breast-fed lamb

The lactic acid bacterium E isolated from the stomach mucus of breast-fed lamb was identified by sequencing of 16S rDNA fragment and species-specific PCR as Lactobacillus reuteri. Its potential antimicrobial activity and ability to modulate immune system in vitro and in vivo was determined. The growth inhibition of potential pathogens decreased from Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella enterica ser. Minnesota to Escherichia coli. The lowest inhibition activity was observed in the case of Candida albicans. The ability of L. reuteri E to modulate biological activities of human and mouse mononuclear cells was estimated in vitro and in vivo, respectively. The production of IL-1 by monocytes in vitro was significantly induced by L. reuteri E (relative activity 2.47). The ability to modulate biological activities of mononuclear cells by living L. reuteri E cells in vitro in comparison to disintegrated L. reuteri E cells in vivo differed. For example lysozyme activity in vitro was inhibited while in vivo was stimulated (relative activities 0.30 and 1.83, respectively). The peroxidase activity in vitro was stimulated while in vivo was inhibited (relative activities 1.53 and 0.17, respectively). Obtained results indicate that L. reuteri E is potential candidate to be used in probiotic preparations for animals and/or human.

Identification and biological activity of potential probiotic bacterium isolated from the stomach mucus of breast-fed lamb

The lactic acid bacterium E isolated from the stomach mucus of breast-fed lamb was identified by sequencing of 16S rDNA fragment and species-specific PCR as Lactobacillus reuteri. Its potential antimicrobial activity and ability to modulate immune system in vitro and in vivo was determined. The growth inhibition of potential pathogens decreased from Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella enterica ser. Minnesota to Escherichia coli. The lowest inhibition activity was observed in the case of Candida albicans. The ability of L. reuteri E to modulate biological activities of human and mouse mononuclear cells was estimated in vitro and in vivo, respectively. The production of IL-1β by monocytes in vitro was significantly induced by L. reuteri E (relative activity 2.47). The ability to modulate biological activities of mononuclear cells by living L. reuteri E cells in vitro in comparison to disintegrated L. reuteri E cells in vivo differed. For example lysozyme activity in vitro was inhibited while in vivo was stimulated (relative activities 0.30 and 1.83, respectively). The peroxidase activity in vitro was stimulated while in vivo was inhibited (relative activities 1.53 and 0.17, respectively). Obtained results indicate that L. reuteri E is potential candidate to be used in probiotic preparations for animals and/or human.

Identification and biological activity of potential probiotic bacterium isolated from the stomach mucus of breast-fed lamb.

The lactic acid bacterium E isolated from the stomach mucus of breast-fed lamb was identified by sequencing of 16S rDNA fragment and species-specific PCR as Lactobacillus reuteri. Its potential antimicrobial activity and ability to modulate immune system in vitro and in vivo was determined. The growth inhibition of potential pathogens decreased from Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella enterica ser. Minnesota to Escherichia coli. The lowest inhibition activity was observed in the case of Candida albicans. The ability of L. reuteri E to modulate biological activities of human and mouse mononuclear cells was estimated in vitro and in vivo, respectively. The production of IL-1ß by monocytes in vitro was significantly induced by L. reuteri E (relative activity 2.47). The ability to modulate biological activities of mononuclear cells by living L. reuteri E cells in vitro in comparison to disintegrated L. reuteri E cells in vivo differed. For example lysozyme activity in vitro was inhibited while in vivo was stimulated (relative activities 0.30 and 1.83, respectively). The peroxidase activity in vitro was stimulated while in vivo was inhibited (relative activities 1.53 and 0.17, respectively). Obtained results indicate that L. reuteri E is potential candidate to be used in probiotic preparations for animals and/or human.

Microbiological and therapeutical aspects of pneumococcal diseases in the Slovak Republic.

Since 1983, multiresistant pneumococcal strains, mostly 14, 23F, 19A serotypes have been reported in Slovakia. A 15-year cooperation of specialists from various fields and the representatives of different institutions revealed that the pneumococcal infection problem in the Slovak Republic remains an important healthcare task. In the prospective multicentric microbiological and clinical analysis of otitis media acuta in children under 5 years, pneumococci were isolated in 45.9%, where 50.8% of strains were intermediate and fully resistant to penicillin and 47.5% resistant to macrolid antibiotics. Invasive pneumococcal infections, mostly meningitis and bacteremias, were observed in two studies. Penicillin resistance was higher in children under 5 years (52.8%), in comparison with other age groups. The dominant serotypes of childhood were 14, 19A and 6A, while in the age group above 65 there was a broad spectrum of serotypes confirmed. Serotype 14 is the most frequent serotype in physiologically sterile liquids and otitis media liquor in the Slovak Republic (Tab. 3, Fig. 5, Ref. 34).

Two cases of severe invasive infections in children caused by Streptococcus pneumoniae serotype 14--case report.

Two cases are presented of severe pneumococcal infections in infants caused by serotype 14 Streptococcus pneumoniae. The first case--meningitis--caused by S. pneumoniae (pneumococcus) with low-level penicillin susceptibility has developed from acute otitis media and resulted in fatal outcome. The second one--an immunocompromised child presenting recurrent otitis and chronic mastoiditis--developed into pneumococcal pneumonia. Both cases demonstrate the extreme importance of a relevant initial treatment of localized pneumococcal infections, preventing the development of generalized infection. Amoxicillin (an oral treatment option in both upper and lower respiratory tract infections caused also by Pneumococcus strains with low-level penicillin susceptibility due to its beneficial pharmacokinetics and pharmacodynamics) was not used in either case.

[Lactobacilli and their probiotic properties]. / Laktobacily a ich probiotické vlastnosti.

Lactobacilli rank among lactic acid bacteria, fermenting saccharides to produce lactic acid. The individual species and strains differ by the profiles of fermented saccharides, but this property is not sufficient for the identification at the species level. The species and strains differ in the 16S rDNA sequence and in the intergenic space between 16S and 23S rDNA, which make their differentiation and identification possible. Some representatives of the genus Lactobacillus are included in probiotic preparations with regard to their beneficial health effects on the human and animal organisms. For example, they are able to interact with the immune system and stimulate it positively, to reduce the manifestations of lactose intolerance by consuming fermented milk products, and to shorten postantibiotic and travellers' diarrhoea. Lactobacilli also produce bacteriocins, antimicrobially active substances.

Effects of LPS, lipid A and polysaccharide from adapted strains of Escherichia coli on human leucocyte activity.

Polysaccharide and lipid A are responsible for the wide-ranging pharmacological activity of bacterial lipopolysaccharides (LPS). The alterations in LPS structure result in various effects on different functions of the target cells. The effects of LPS substructures, the polysaccharide (P) and lipid A (L) from E. coli on the innate mechanisms of human leucocytes were examined and compared in this study. Incubation of leucocytes with LPS and L and P analogues (1 and 100 microg/ml) enhanced their biological activity in dependence on their structure. These results showed that LPS was a less active immunomodulator of leucocytes than L and P analogues isolated from E. coli strains adapted to antimicrobial agents.

NO-synthase inhibitors provide influence on protective effect of modified endotoxine diphosphoryl lipid A in a rat heart model of ischemic-reperfusion injury.

The present study was designed to assess whether a protective effect of the modified diphosphoryl lipid A (modLA) against myocardial ischemia-reperfusion injury (IRI) in rats can be related to the mechanism involving inducible nitric oxide synthase (iNOS). Pre-treatment with modLA significantly reduced the duration of both ventricular tachycardia (p < 0.01) and ventricular fibrillation (p < 0.001) compared to controls. Under these conditions the incidence of animal death was reduced (p < 0.05). The beneficial effect of modLA was markedly attenuated by the prior administration of selective iNOS inhibitor S-methylisothiourea (SMT). In this animal group, mortality was significantly increased (p < 0.01) partially in consequence of sustained ventricular arrhythmias. These results indicate that induction of iNOS can be responsible for cardioprotection of modLA.

Persistent suppression of phagocytosis after prolonged administration of local anesthetics carbisocaine and heptacaine in mouse leukocytes.

Effects of local anesthetics heptacaine and carbisocaine on mouse peritoneal macrophages after 5 days of peritoneal administration were observed. Both compounds in a daily dose 50 mg/kg caused significant decrease in phagocytic activity and index of phagocytosis (number of ingested particles). No significant effects on phagocytosis were observed when carbisocaine was administrated in a daily dose 5 mg/kg. No significant changes in weight gain and number of peritoneal macrophages were observed in all groups.

[Anticomplement activity of saponins from Cynara cardunculus L]. / Antikomplementová aktivita saponínov z Cynara cardunculus L.

Triterpene saponins of ursane and oleane type isolated from the involucral bracts of Cynara cardunculus L. (Asteraceae) were investigated in in vitro assays for their activity on the complement system. The anticomplementary activity of bidesmosidic saponins on the classical pathway activation of the complement was higher than the activity of monodesmosidic saponins, but esterification of the carboxylic group of glucuronic acid in the sugar residue resulted in a significant decrease in anticomplementary activity.

Activation of human leukocytes by lipid A from E. coli strains adapted to quaternary ammonium salt and amine oxide.

The immunomodulatory activities of monophosphoryl lipid A (MLA) and diphosphoryl lipid A analogues obtained from the sensitive strain of E. coli and from the resistant strains adapted to a quaternary ammonium salt and an amine oxide were compared. All analogues considerably stimulated the activity of human leukocytes although the analogue from the sensitive strain at a higher concentration significantly suppressed phagocytosis. The MLA analogue exhibited a suppressive effect on the microbicidal activity of human leukocytes against E. coli and the peroxidase activity. Adaptation of bacteria to amphiphilic antimicrobial compounds, which is accompanied by chemical changes in their lipid A, only slightly reduced their immunomodulatory activity when compared with the analogue from the sensitive strain. On the other hand, the diphosphoryl analogues were less active than MLA.

Comparative study of disintegrated cells influence of Staphylococcus aureus, Escherichia coli and Candida albicans on human and mouse immune mechanisms.

The study presents comparison of immunomodulatory effects of Staphylococcus aureus, Escherichia coli and Candida albicans disintegrated cells on selected immune mechanisms of human and mouse leukocytes. We measured their phagocytic activity, phagocytic index and microbicidal activity against Staphylococcus aureus, Escherichia coli and Candida albicans cells as well as peroxidase and lysozyme activities of human and mouse leukocytes. Our results revealed predominantly inhibitory effect of disintegrated microorganisms on nonspecific immune functions of human leukocytes, but mainly stimulatory effect on mouse leukocytes monitored immune functions.

[Anticomplement activity of Mahonia aquifolium bisbenzylisoquinoline alkaloids and berberine extract]. / Stúdium antikomplementovej aktivity extraktu Mahonia aquifolium bisbenzylizochinolínových alkaloidov a berberínu.

Crude extracts obtained from the stem bark of Mahonia aquifolium have been investigated as to the chemical composition and anticomplementary activity. The results show that their anticomplementary activity is mainly due to the alkaloid components. Especially the BBI alkaloid fraction and berberine showed a strong inhibitory effect on CH50 total hemolytic complement assay. The crude extract of M. aquifolium was less active than berberine or the fraction BBI alkaloids. The results indicate that the fraction of BBI alkaloids and berberine largely account for the immunomodulatory activity of the crude extract of M. aquifolium.

Immunomodulatory activity of amphiphilic antimicrobials on mouse macrophages.

A quaternary ammonium salt (1-methyldodecyl)-trimethylammonium iodide, a structurally analogous amine oxide (1-methyldodecyl)-dimethylamine-N-oxide and the amine oxide lacking long alkyl chain trimethylamine-N-oxide were tested for their immunomodulatory activity. Inbred mice strain C57/BL6 were pretreated for 7 days by the compounds under study. The activity of elicited peritoneal macrophages was also tested. Both compounds have a long alkyl chain. In concentrations of 10(-6) M there was a significant increase of the phagocytic, candidacidal and lysozyme activities of the cells. We also observed a suppressed peroxidase activity. The colicidal activity of both the peritoneal and spleen cells were not affected. The amine oxide lacking the long alkyl chain has the same effect at high concentration. A similarity between the effects of the amphiphilic compounds on the macrophages and their antimicrobial efficacy elicits the conclusion that both activities are caused by their ability to interact with the cell membranes.

Immunomodulating activity of Echinacea gloriosa L., Echinacea angustifolia DC. and Rudbeckia speciosa Wenderoth ethanol-water extracts.

The effect of the ethanol-water extracts of Echinacea gloriosa L., Echinacea angustifolia DC. and Rudbeckia speciosa Wenderoth on immunological system of inbred mice was investigated. The extract of the root of Rudbeckia speciosa had the highest immunostimulatory activity.

[Testing for immunomodulating effects of ethanol-water extracts of the above-ground parts of the plants Echinaceae (Moench) and Rudbeckia L]. / Testovanie imunomodulacných úcinkov etanolovo-vodných extraktov z nadzemných castí rastlín rodu Echinacea Moench a Rudbeckia L.

Ethanolic extracts of aerial parts of the plants Echinacea angustifolia DC, Echinacea purpurea L. (Moench), Rudbeckia fulgida var. sullivantii, Boyton et Beadle, and Rudbeckia speciosa Wenderoth show immunomodulating activity. The mice were treated in vivo for 5 days and the activity was tested for and observed on day 7. An immunostimulatory effect was observed on the phagocytic, metabolic and bactericidal activities of peritoneal macrophages. The ethanolic extracts of both Echinacea plants also increased the total weight of the spleens as compared to the effect of the Rudbeckia plants and the control group which received saline.

[Immunomodulating activity of ethanol-water extracts of the roots of Echinacea gloriosa L., Echinacea angustifolia DC. and Rudbeckia speciosa Wenderoth tested on the immune system in C57BL6 inbred mice]. / Imunomodulacná aktivita etanolovo-vodných extraktov z korennov Echinacea gloriosa L., Echinacea angustifolia DC. a Rudbeckia speciosa Wenderoth testovaná na imunitnom systéme inbredných mysí C57BL6.

The ethanolic extract from the roots Echinacea gloriosa L. (Moench), Echinacea angustifolia DC. and Rudbeckia speciosa Wenderoth shows immunomodulating activity. It was seen on the seventh day after five days of in vivo treatment of mice. The most marked immunostimulatory effect was observed on the lysosomal and peroxidal activity of peritoneal macrophages, and splenic cells after in vivo treatment with the ethanolic extract of the roots of R. speciosa Wenderoth.

Immunochemical properties of Vibrio cholerae LPS.

Immunochemical analysis of LPS isolated from Vibrio cholerae O1 and non O1 showed that this macromolecular complex shares common antigenic epitopes in the sugar moiety. The epitopes can be detected after mild alkaline hydrolysis of LPS in vitro. Membrane-associating activity of both O1 and non O1 LPS did not indicate any differences of the species.