Studies of viomycin, an anti-tuberculosis antibiotic: copper(ii) coordination, DNA degradation and the impact on delta ribozyme cleavage activity.

Viomycin is a basic peptide antibiotic, which is among the most effective agents against multidrug-resistant tuberculosis. In this paper we provide the characteristics of its acid base properties, coordination preferences towards the Cu(ii) ions, as well as the reactivity of the resulting complexes against plasmid DNA and HDV ribozyme. Careful coordination studies throughout the wide pH range allow for the characterisation of all the Cu(ii)-viomycin complex species. The assignment of proton chemical shifts was achieved by NMR experiments, while the DTF level of theory was applied to support molecular structures of the studied complexes. The experiments with the plasmid DNA reveal that at the physiological levels of hydrogen peroxide the Cu(ii)-viomycin complex is more aggressive against DNA than uncomplexed metal ions. Moreover, the degradation of DNA by viomycin can be carried out without the presence of transition metal ions. In the studies of antigenomic delta ribozyme catalytic activity, viomycin and its complex are shown to modulate the ribozyme functioning. The molecular modelling approach allows the indication of two different locations of viomycin binding sites to the ribozyme.

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.

Effect of a single bout of resistance exercise on arterial stiffness following a high-fat meal.

Consumption of a high-fat meal (HFM) causes postprandial lipemia and vascular dysfunction. Acute resistance exercise (RE) alone may also have a negative effect on vascular function. The purpose of this study was to measure arterial stiffness and postprandial lipemia after a HFM with or without acute RE. 9 recreationally active men (age 24±5 years, BMI 25±3 kg/m2) completed both: (1) HFM alone and (2) HFM+RE in a randomized order. Pulse wave velocity (PWV) from carotid to femoral artery and carotid to radial artery were used as measures of central/aortic and peripheral arterial stiffness, respectively. Circulating triglycerides (TRG) were obtained from finger stick samples as a marker of lipemia. There was a significant condition-by-time interaction for TRG (p<0.05). TRG levels increased significantly following both conditions with a significantly attenuated increase following HFM+RE (p<0.05). There was a significant condition-by-time interaction for peripheral PWV as this parameter increased following HFM, but decreased following HFM+RE (p=0.021). Central PWV did not change with HFM or HFM+RE (p>0.05). Following a HFM, acute RE attenuates postprandial lipemia and improves peripheral arterial stiffness without having a negative effect on central arterial stiffness.

Applicability of endoscopic examination as a diagnostic approach in urinary tract ailments in dogs.

The purpose of this study was to evaluate the applicability of endoscopic examination as a diagnostic method in urethra and bladder ailments in dogs. The experiment involved 60 dogs of different breeds and gender, weighing from 5 to 65 kg, aged between 7 months and 12 years. The dogs were divided into two groups. Group I consisted of clinically healthy animals on which all the lower urinary tract endoscopic examinations described in the literature as well as biopsies of mucosa were performed. Group II consisted of clinical patients, on which endoscopic diagnostic examination was conducted and, on some of them, biopsies for histopathologic examination were made. The results of the experiment suggest that the lower urinary track endoscopy is a suitable diagnostic technique for recognition and differentiation of mucosa inflammation, establishment of causes of haematuria, diagnostics of neoplasms and detection of the existence of an ectopic ureter. Biopsy and histopathologic examination improved the diagnostic value of endoscopy. The techniques tested in our study, inclusive of more invasive method like PPC and cystoscopy on males preceded by the perineal urethrotomy, enable doctors to perform the lower urinary tract endoscopic examination on dogs, especially up to 5 kg. regardless of their gender.

Laparoscopic cryptorchidectomy in dogs --report of 15 cases.

The study aimed at determining the usability of laparoscopic cryptorchidectomy in treating cases of simple cryptorchidism and neoplastic testes in dogs. The presence of one or both testes inside the peritoneal cavity was confirmed in this study by an ultrasonographic examination employing the use of a 6.5 MHz convex head. The surgeries were conducted on a group of 15 dogs. In 12 subjects a one-sided cryptorchidism was found, 9 of which had right-sided cryptorchidism. In 3 dogs, both-sided cryptorchidism was observed. In 3 cases, an increase in the size of the abdominal testes with the symptoms of hyperestrogenism and feminisation raised a suspicion of neoplastic changes (SCT) taking place, which was confirmed by a histopathological examination. A laparoscopic cryptorchidectomy with intracorporal ligation was performed in all the subjects. The spermatic cord and ductus deferens were occluded with a ligature of an absorbable material, and a manually applied knot.

Persistent skin colonization with Staphylococcus aureus in atopic dermatitis: relationship to clinical and immunological parameters.

BACKGROUND: Staphylococcus aureus has important implications for the pathogenesis of atopic dermatitis (AD). In some patients S. aureus can be eradicated from the skin during anti-inflammatory treatment, while in others bacterial colonization is persistent. Potential mechanisms and features of these two distinct groups of patients are not known. OBJECTIVE: Accordingly, we studied relationships between the ability to eliminate S. aureus during an anti-inflammatory treatment and selected clinical and immunological features. METHODS: Quantitative assessment of S. aureus on the skin, in nasal vestibule and throat, serum IgE levels, CD4/CD8 T-cell ratio, lymphocyte proliferation and phagocyte oxidative burst were determined during the exacerbation and after 4 and 12 weeks of the treatment using topical steroid and oral antihistamine in 34 patients with AD. RESULTS: S. aureus was found on the skin of all 34 patients during exacerbation. Disease severity scoring of atopic dermatitis (SCORAD) correlated with the density of bacteria. Treatment with oral antihistamine and topical steroid resulted in a significant alleviation of symptoms, which correlated with the elimination of S. aureus from the skin in 70% of patients. In the remaining 30% of patients, dense (more than 10(10)/cm2) S. aureus skin colonization, persisted despite the treatment. Patients with persistent S. aureus presented with higher serum IgE levels, lower lymphocyte proliferation in response to staphylococcal enterotoxin B, phytohaemagluttinin and anti-CD3. Persistence of S. aureus was more common in men. CONCLUSIONS: Patients with AD differ in the ability to clear S. aureus from the skin during anti-inflammatory treatment, which appears to be related to the abnormalities in immunological parameters. Local antibiotic therapy should be considered only in patients with persistent S. aureus colonization.

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.

[Chronic inflammation of the colon: a diagnostic problem]. / Przewlekle odczyny zapalne jelita grubego--problem diagnostyczny.

Chronic inflammation of the colon and rectum with associated diarrhea has become more difficult to diagnose recently. Many etiological factors and different therapeutic outlines need to be carefully discussed and summarized. Based on most recent literature, authors present current data on etiology and specific (case-depended) diagnostic procedures in colitis and proctocolitis.

Two allelic forms of the aureolysin gene (aur) within Staphylococcus aureus.

Proteinases of Staphylococcus aureus are emerging as potential virulence factors which may be involved in the pathogenecity of staphylococcal diseases. We describe here the structure of the gene encoding the metalloproteinase referred to as aureolysin. This gene occurs in two allelic forms and is strongly conserved among S. aureus strains, implying the possibility that the proteinase may have important housekeeping functions.

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.

Absence of enhanced sympathoadrenal activity and behaviorally evoked cardiovascular reactivity among offspring of hypertensives.

To determine whether offspring of hypertensives show enhanced sympathetic nervous system activity, we evaluated several indices of sympathoadrenal activation and cardiovascular responsiveness to behavioral stimuli among 90 normotensive, young adult men having either one or two hypertensive parents (PH+(-), PH++) or normotensive parents only (PH--) (n = 30/group). Measurements included heart rate (HR) and blood pressure (BP) reactions to three mental stressors (the Stroop test, mental arithmetic, mirror tracing), a cold pressor test, postural adjustment (60 degrees upright tilt), isometric exercise and bicycle ergometry, as well as the 24-h excretion of catecholamines (epinephrine [E], norepinephrine [NE]) and venous plasma catecholamine concentrations, both at rest (seated and supine) and in response to the Stroop test and upright tilt. The three groups did not differ in age, education, body mass index (BMI), estimated aerobic fitness, resting HR, cardiac preejection period (PEP) and PEP:LVET (left ventricular ejection time) ratio, 24-h Na or K excretion, or fasting lipids, insulin or plasma renin activity. Resting systolic and diastolic BP varied as a function of parental hypertension, and were significantly higher in PH++ than among PH-- subjects (P < .05). No significant group difference was observed on any measure of plasma or urinary catecholamines, nor did offspring of hypertensives (PH++ or PH+-) showed greater HR or BP reactions than PH-- subjects to any of the several laboratory challenges. In sum, we find no evidence of enhanced sympathetic activity or heightened cardiovascular responsiveness among normotensive young adults who are familially predisposed to essential hypertension.

Differential effects of chlorination of bacteria on their capacity to generate NO, TNF-alpha and IL-6 in macrophages.

Activated rodent macrophages produce high amounts of nitric oxide (NO). NO as a tumoricidal and defence molecule against intracellular parasites is commonly accepted. However, its role as an obligatory killing factor for extracellular bacteria is controversial. In the present study we stimulated murine peritoneal macrophages by heat-killed bacteria (Staphylococcus aureus, S. epidermidis and Escherichia coli). In some groups bacteria were pretreated with HOCl, to replace the chlorinating system in activated neutrophils that operates as a bactericidal system in vivo. High levels of NO, tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were detected after stimulation by all non-chlorinated bacteria strains tested. However, after chlorination Gram-positive bacteria lost their ability to induce NO and TNF-alpha, whereas phagocytosis and IL-6 production were not affected by chlorination.

[Bacteriology of acne changes]. / Bakteriologia zmian tradzikowych.

In this work it was demonstrated that the main microorganisms which cause, together with other, the generation of inflammatory changes are anaerobic bacteria from the genus Propionibacterium. P. acnes type I causes mild course acne changes while serious changes are generated by P. granulosum type II. It was observed that a high percentage of all tested Propionibacterium strains showed susceptibility to antibiotics but it did not reach 100%. In the treatment of acne changes all agents which could be conducive to generating them should be taken into consideration. The therapy should be complex and applied with regard to the course of disease of each individual patient.

[Clinical evaluation of serious forms of acne treated with autovaccine]. / Ocena kliniczna ciezkich postaci tradzika leczonych autoszczepionka.

In this paper a clinical assessment of the treatment of serious forms of acne vulgaris with the use of autovaccines containing Propionibacterium strains was presented. It was shown that the associative therapy including parallel administration of both autovaccine and the antibiotic is remarkably efficacious.

[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.