Comparative Analysis of Microbicidal and Anti-inflammatory Properties of Novel Taurine Bromamine Derivatives and Bromamine T.

Taurine, the most abundant free amino acid in leukocyte cytosol traps hypohalous acids (HOCl and HOBr) to produce N-chlorotaurine (taurine chloramine, NCT and N-bromotaurine (taurine bromamine, Tau-NHBr,) respectively. Both haloamines show anti-inflammatory and antimicrobial properties. However, the therapeutic applicability of Tau-NHBr is limited due to its relatively poor stability. To overcome this disadvantage, we have synthesized the stable N-bromotaurine compounds N-monobromo-2,2-dimethyltaurine (Br-612) and N-dibromo-2,2-dimethyltaurine (Br-422). The aim of this study was to compare anti-inflammatory and microbicidal properties of Br-612 and Br-422 with that of Tau-NHBr and bromamine T (BAT). We have shown that all the tested compounds show similar anti-inflammatory properties. Importantly, the stable N-bromotaurine compounds exerted even stronger microbicidal activity than Tau-NHBr. Finally, for the purpose of topical application of these compounds we have developed a carbomer-based bioadhesive solid dosage form of BAT and Br-612, featuring sustained release of the active substance.

Clustering of Staphylococcus aureus bovine mastitis strains from regions of Central-Eastern Poland based on their biochemical and genetic characteristics.

Staphylococcus aureus strains were isolated from mastitic milk of cows with infected mammary glands. The animals were living in 12 different farms near Lublin, in Central-Eastern Poland. A biochemical identification method based on enzymatic assay was performed, followed by haemolytic and proteolytic tests. PCR-RFLP targeted on the gap gene allowed the genetic identification of strains at the species level and verified phenotypic identification results. A molecular typing method using triplex PCR was performed to recognize the genetic similarity of the analyzed strains. DNA microarray hybridization (StaphyType, Alere Technologies) was used for detection of antibiotic resistance and virulence associated markers. The results obtained indicate high genetic similarity in strains isolated from the same sites. High genetic similarities were also detected between strains isolated from cows from different farms of the same region. A slightly lower similarity was noted however, in strains from various regions indicating that the strains are herd specific and that the cow's infections caused by S. aureus were of a clonal character. In 21 representative isolates selected for DNA-microarray testing, only fosfomycin (fosB) and penicillin resistance markers (blaZ, blaI, blaR) were detected. The presence of genes coding for haemolysins (lukF, lukS, hlgA, hla, hld, hlb), proteases (aur, sspA, sspB, sspP), enterotoxins (entA, entD, entG, entI, entJ, entM, entN, entO, entR, entU, egc-cluster), adhesins (icaA, icaC, icaD, bbp, clfA, clfB, fib, fnbA, map, vwb) or immune evasion proteins (scn, chp, sak) was common and, with exceptions, matched triplex PCR-defined clusters.

Is there a role of taurine bromamine in inflammation? Interactive effects with nitrite and hydrogen peroxide.

OBJECTIVE AND DESIGN: The myeloperoxidase system of neutrophils generates chlorinating and brominating oxidants in vivo. The major haloamines of the system are taurine chloramine (TauCl) and taurine bromamine (TauBr). It has been demonstrated in vitro that TauCl exerts both antiinflammatory and anti-bacterial properties. Much less is known about TauBr. The present study was conducted to compare bactericidal and immunoregulatory capacity of TauBr with that of the major chlorinating oxidants: HOCl and TauCl. Moreover, the effect of nitrites and H(2)O(2) on TauBr activity was investigated. MATERIALS: TauBr was prepared by reaction of HOBr with taurine. The reaction was monitored by UV absorption spectra. METHODS: Bactericidal activity of TauBr, TauCl and HOCl was tested by incubation of E. coli with the compounds and determined by the pour-plate method. To test the anti-inflammatory activity the compounds were incubated with LPS and IFN-gamma stimulated murine peritoneal macrophages. The production of following mediators was measured: nitrites by Griess reaction; TNF-alpha, IL-6, IL-10, IL-12p40 using capture ELISA. In some experiments the compounds were incubated with either nitrites or H(2)O(2). RESULTS: In our experimental set-up TauBr and HOCl exerted strong bactericidal effects on E. coli (MBC = 110 microM and 8 microM, respectively), while TauCl (< 1000 microM) did not kill test bacteria. However, both, TauBr and TauCl, at noncytotoxic concentrations (< 300 microM) inhibited the cytokine and nitric oxide production by macrophages. H(2)O(2) completely abolished the biological activities of TauBr but not those of TauCl. Nitrites did not affect any activity of TauBr or TauCl while they diminished the HOCl(-) mediated bacterial killing. CONCLUSION: TauBr, despite very low concentration of Br(-) in body fluids, may support TauCl and HOCl in the regulation of inflammatory response and in killing of bacteria by neutrophils. However, TauBr activity in vivo will depend on the presence of H(2)O(2) and possible other mediators of inflammation which can compete with target molecules for TauBr.

Proteolytic activity of Staphylococcus aureus strains isolated from the colonized skin of patients with acute-phase atopic dermatitis.

Staphylococcus aureus strains isolated from the colonized skin lesions of 26 patients with acute-phase atopic dermatitis were reported to produce various extracellular proteolytic enzymes. Using the skim-milk-agar culture plating method, it was shown that 97% of the strains (65 of 67 examined) produced proteolytic activity, with 61% (42 strains) producing activity comparable to that of the proteolytically hyperactive reference strain Staphylococcus aureus V8. This observation was confirmed by azocasein degradation with culture supernatants, which indicated that 91% of the strains produced extracellular proteinases and 43% exceeded the 2% activity threshold of the reference strain. Control strains were isolated from the nose vestibules of 18 healthy carriers; the proteolytic activity of these strains never exceeded 2.5% of the activity of the reference strain. In 54% of the patients examined ( n=14), the activity of the strains was higher than that determined for the isolates from the control group. The combined use of assays incorporating azocasein and a synthetic chromogenic substrate, N-CBZ-Phe-Leu-Glu- pNA, showed that two staphylococcal enzymes, Staphylococcus aureus metalloproteinase (SAMP) and Staphylococcus aureus serine proteinase (SASP), contributed to the total proteolytic activity released by the strains examined. The contribution of each of the two enzymes varied greatly between different isolates. The undamaged skin of the patients was not colonized with Staphylococcus aureus. The presence of several strains with atypical proteinase characteristics was also reported, suggesting the possible involvement of enzymes other than serine- and metallo-proteinases in the proteolytic activity of Staphylococcus aureus. Taken together, the results of the study imply that staphylococcal proteinases may contribute to the pathogenicity of atopic dermatitis.

Synthesis, spectral and antimicrobial properties of 5-chloroarylidene aromatic derivatives of imidazoline-4-one.

The synthesis of new chloro-benzylidene substituted derivatives of hydantoin and their antimicrobial activity is reported. The structure-activity relationships showed that the antibacterial effect of investigated compounds depends on the distance of the phenyl ring from the amine residue and the kind of substitutes on the phenyl ring. In the investigated group of derivatives, 5-(2-chlorobenzylidene)-2-(4-fluorobenzylamine)-imidazoline-4-one and 5-(2-chlorobenzylidene)-2-(2-phenylethylamine)-imidazoline-4-one showed the best antibacterial activity against Moraxella catarrhalis.

[Staphylococcus strains isolated from cases of osteomyelitic patients qualifying for autovaccine therapy]. / Szczepy Staphylococcus aureus izolowane z przypadków zapalen kosci u pacjentów kwalifikowanych do leczenia autoszczepionka.

Staphylococcus aureus strains isolated from osteomyelitic were analysed biochemically and phage-typed. All examined strains were classified to the following biochemical groups: biotype I 6,332,153, biotype II 6,732,151, biotype III 6,732,151, biotype IV 6,732,150. Among the strains the following types were frequent: 3A, 3C, 55, 71 and 96.

[Bacterial flora of respiratory tract infections]. / Flora bakteryjna w infekcjach dróg oddechowych.

In this paper the bacterial flora of respiratory tract diseases of children and adults was described. Significant differences in the frequency of isolation of various species of bacteria in connection with the age and the clinical form of disease were observed. Haemophilus influenzae was isolated from the accessory sinuses of the nose of children with significantly higher frequency than from adults. This microorganism occurred as the only etiological agent in more than 50% of cases.

[Characteristics of lactobacillus species strains isolated from reproductive organs in various clinical cases]. / Charakterystyka paleczek z rodzaju Lactobacillus izolowanych z dróg rodnych, z róznych przypadków klinicznych.

The study was aimed at detection of presence of Lactobacillus species bacteria in 167 samples of material derived from vagina of patients with cervix uteri (22 samples), chronic vaginosis (100 samples) and from women after treatment of inflammatory diseases (45 samples). Out of 100 isolated strains of Lactobacillus only 13 belonged to L. acidophilus. They were mainly present (10 strains) in patients after effective treatment of inflammatory diseases of reproductive organs. Remaining strains belonged to L. jensenii (18 strains) and Lactobacillus sp. (69 strains) and did not cleave glycogen. This was detected by quantitative determinations of lactic acid in cultures of these bacteria in the fluid media containing 0.25% of glycogen. It seems that treatment of bacterial infections of reproductive organs should be concentrated on not only elimination of pathogenic bacteria, but also on noneffective strains from Lactobacillus species (strains unable to cleave glycogen with production of lactic acid).

[Gardnerella vaginalis in infections of reproductive organs]. / Gardnerella vaginalis w infekcjach dróg rodnych.

The study was aimed at multidirectional studies on bacteria isolated from smears from vagina and cervix uteri in 226 patients with inflammatory states of their reproductive organs. Most frequently isolated aerobic bacteria were Gram-negative--27%, mainly E. coli, and Enterococcus faecalis--in 18% of cases. Anaerobic bacteria were isolated in 81% of cases: Gardnerella vaginalis was isolated in 28% and Lactobacillus in 53% of cases. Other anaerobic bacteria were: Peptococcus asaccharolyticus (15.5%), Streptococcus sp. (15.9%), and Bacteroides melaninogenicus (14.1%). Gardnerella vaginalis was most frequently found in chronic cases of vaginosis (41.7%). All strains of G. vaginalis were susceptible to cefotaxime, while 15-40% of them were resistant to gentamycin, tetracycline and metronidazole.

[The role of haemophilus in respiratory tract diseases in children from Silesian Center of Children Rehabilitation in Rabka]. / Rola Haemophilus w schorzeniach dróg oddechowych u dzieci ze Slaskiego Osrodka Rehabilitacji Dzieci w Rabce.

In this work the frequency of isolation of rods the genus Haemophilus from children suffering from various clinical sharpes of respiratory tract diseases. All isolated strains were tested in respect of species, biotype, the ability of adhesion to oral and laryngeal epithelia and their susceptibility to routinely applied antibiotics. A strong correlation between the species and biotype of rods from the genus Haemophilus and clinical shape of respiratory tract diseases was found. It was observed that the Haemophilus rods show differences in the ability of adhesion to oral and laryngeal epithelia. Only 52% of the isolated strains were susceptible to bactrim and 88% to tetracycline.

[Anaerobic bacteria detected in inflammatory conditions of the respiratory tract]. / Bakterie beztlenowe wykrywane w stanach zapalnych ukladu oddechowego.

In this study a participation of anaerobic bacteria in respiratory tract diseases is presented. Bronchial washings collected by ++fibrobronchoscope constituted material for the study. Immediately after collection the material was plated onto two media for aerobic bacteria (hemomedium) and anaerobic bacteria (anaeromedium). Then, the samples were centrifuged and a sediment was plated on solid media suitable for aerobic and anaerobic bacteria. Bacterial anaerobic isolates were identified by using API 20E and their sensitivity to antibiotics was tested. From the material described above the most frequently isolated anaerobic bacteria were such as: Streptococcus intermedius, Bacteroides melaninogenicus, Veilonella sp. Among aerobic bacteria the most frequently isolated were Gram-negative rods, Streptococcus faecalis, Branhamella catarrhalis. It is worth to underline that in about 25% of cases anaerobic bacteria were the only isolates.

Erythromycin and penetrating agents containing ointment in acne vulgaris therapy.

An ointment containing penetrating agents and erythromycin was devised. The rate of penetration of the antibiotic through the skin and the ointment efficacy in treatment of acne changes caused by Propionibacterium strains in animals were evaluated.