Multidirectional characterisation of chemical composition and health-promoting potential of Rosa rugosa hips.

Rugosa rose provides one of the largest hips frequently used in the preparation of pharmaceutical and food products. The aim of work was to conduct multidirectional study of biological activity and chemical composition of Rosa rugosa hips. Antiradical, cytotoxic (against cervical and breast cancer cell lines), antibacterial (against eight bacterial strains) and antifungal potential of the species in question was evaluated. Total contents of phenolics, phenolic acids, flavonoids, tannins, carotenoids and ascorbic acid were determined. LC-ESI-MS/MS analysis was performed in order to investigate closely phenolic acids and flavonoid glycosides. As a result, interesting selective cytotoxic effects on cervical (HeLa) and breast cancer (T47D) cell lines, significant antiradical activity (EC50 2.45 mg mg-1 DPPH•) and moderate antimicrobial potential (MIC 0.625-1.25 mg mL-1) were observed. Nine phenolic acids and 11 flavonoid glycosides were qualitatively and quantitatively determined, including 7 compounds previously not reported in R. rugosa hips.

Haemophilus parainfluenzae as a marker of the upper respiratory tract microbiota changes under the influence of preoperative prophylaxis with or without postoperative treatment in patients with lung cancer.

BACKGROUND: Haemophili are representative microbiota of the upper respiratory tract. The aim of this study was to assess the effects of perioperative antimicrobial prophylaxis and/or postoperative treatment on Haemophilus parainfluenzae prevalence, and antimicrobial sensitivity in short-term hospitalized patients with lung cancer who underwent surgery. RESULTS: Samples were collected from 30 short-term hospitalized patients with lung cancer and from 65 healthy people. The nasal and throat specimens were taken twice from each patient: before (EI, Examination I), on the fourth/fifth day (EII, Examination II) after surgery, and once from healthy people. The isolates identification and antimicrobial susceptibility were detected by routine diagnostic methods. H. parainfluenzae was found in throat specimens of 42/65 (64.6 %) healthy people, while in 19/30 (63.3 %) lung cancer patients in EI (p = 0.6203) and in 13/30 (43.3 %) ones in EII (p = 0.0106). Neither the disease itself nor short-term hospitalization with perioperative prophylaxis alone affected H. parainfluenzae prevalence in EII, while perioperative prophylaxis with postoperative treatment significantly decreased its colonization in EII. The differences in the number of patients colonized by Candida spp. in EI and in EII were observed (p = 0.0082).Totally, 23/58 (39.7 %) of H. parainfluenzae isolates were resistant mainly to beta-lactams; among 11 ampicillin-resistant isolates only 3 were beta-lactamase positive. CONCLUSIONS: The antimicrobial perioperative prophylaxis together with postoperative treatment may disturb the composition of the airways microbiota represented by H. parainfluenzae, in addition to selecting the resistant strains of bacteria and promoting yeasts colonization in lung cancer patients undergoing surgery.

Antibacterial, Antiradical Potential and Phenolic Compounds of Thirty-One Polish Mushrooms.

BACKGROUND: Among many sources of natural bioactive substances, mushrooms constitute a huge and almost unexplored group. Fungal compounds have been repeatedly reported to exert biological effects which have prompted their use in pharmaceutical and cosmetic industry. Therefore, the aim of this study was analysis of chemical composition and biological activity of 31 wild growing mushroom species (including saprophytic and parasitic) from Poland. METHODS: Qualitative and quantitative LC-ESI-MS/MS analysis of fourteen phenolic acids in the mushrooms analysed was performed. Moreover, total phenolic content was determined by the modified Folin-Ciocalteau method. Antioxidative activity of ethanolic extracts towards DPPH• free radical was examined. Antibacterial activity against Gram-positive (S. epidermidis, S. aureus, B. subtilis, M. luteus) and Gram-negative (E. coli, K. pneumoniae, P. aeruginosa, P. mirabilis) microbial strains was analyzed. RESULTS: As a result, the first such broad report on polyphenolic composition, antiradical and antimicrobial potential of wild growing Polish mushrooms was developed. Mushroom extracts were found to contain both benzoic (protocatechuic, 4-OH-benzoic, vanillic, syringic) and cinnamic acid derivatives (caffeic, p-coumaric, ferulic). Total phenolic content in mushrooms ranged between 2.79 and 53.13 mg gallic acid equivalent /g of dried extract in Trichaptum fuscoviolaceum and Fomes fomentarius, respectively. Fungi showed much differentiated antiradical activity, from highly active F. fomentarius to poorly effective Russula fragilis (IC50 1.39 to 120.54 mg per mg DPPH•, respectively). A quite considerable relationship between phenolic content and antiradical activity has been demonstrated. Mushrooms varied widely in antimicrobial potential (MIC from 0.156 to 5 mg/ml). Generally, a slightly higher activity against Gram-positive than Gram-negative strains was observed. This is the first study concerning the chemical composition and biological activity of the majority of investigated species.

Cytotoxic, antioxidant, antimicrobial properties and chemical composition of rose petals.

BACKGROUND: Rosa rugosa petals are used for production of teas, jams, wines and juices. Despite the wide availability of rose cultivars, comprehensive information on petal chemical composition and healthful properties is still lacking. Therefore, the aim of this study was analysis of cytotoxic, antioxidant and antimicrobial activity and chemical composition of rugosa rose petals. RESULTS: Petals of R. rugosa were evaluated for their cytotoxic effect against cervical (HeLa) and breast cancer (T47D) cell lines and for antiradical activity (with DPPH•). As a result, significant cytotoxic (up to 100% of dead cells) and antiradical properties (IC50 1.33-0.08 mg mg⁻¹ DPPH•) were demonstrated. Moreover, notable antimicrobial activity against eight bacterial (i.e. Staphylococcus. epidermidis, S. aureus, Bacillus subtilis, Micrococcus luteus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis) and two yeast strains (Candida. albicans, C. parapsilosis) was shown. Total phenolic, flavonoid, phenolic acid, tannin, carotenoid and polysaccharide content in petals was determined using spectrophotometric methods. Liquid chromatography-electrospray ionization-tandem mass spectrometry was used to thoroughly analyze phenolic acids and flavonoid glycosides in the methanolic extract and fractions obtained after its separation. Five phenolic acids and six flavonoids previously not reported in the plant material were identified. CONCLUSION: This is the first such detailed report on chemical composition and biological activity of R. rugosa petals.

Quantitative analysis of biofilm formed on vascular prostheses by Staphylococcus epidermidis with different ica and aap genetic status.

OBJECTIVES: This study aims to examine biofilm formed on vascular prostheses by Staphylococcus epidermidis with different ica and aap genetic status, and to evaluate the effect of antibiotic-modified prostheses on bacterial colonization. METHODS: Biofilm formation was determined using fluorescence microscopy imaging. Quantitative analysis was conducted using the biofilm coverage ratio (BCR) calculations. RESULTS: Our investigations prove that the BCR method with fluorescent dye enabled an accurate assessment of biofilm coverage and comparison of the obtained results. The ica+ aap+ strains formed a biofilm on all of the examined vascular prostheses. Uni-Graft(®) modified with covalently immobilized amikacin was effective in preventing bacterial adherence. CONCLUSIONS: Molecular biology techniques combined with phenotype studies give a broad insight into biofilm formation mechanisms. On the other hand, fluorescence microscopy imaging along with BCR calculations are reliable and simple tools to quantitatively estimate biofilm formation, as well as the effectiveness of antimicrobial prosthesis modification.

Phenolic acids content, antioxidant and antimicrobial activity of Ligusticum mutellina L.

A simple HPLC method has been used for separation and quantitative analysis of the phenolic acids in the methanolic extracts of Ligusticum mutellina aerial parts. Chlorogenic acid was the predominant phenolic acid. Additionally, gallic, p-OH-benzoic, caffeic, p-coumaric and ferulic acids were identified. Moderate antibacterial and antifungal activity (MIC = 1.25-2.5 mg mL(-1)) was observed for the methanol extract of L. mutellina herb received from plants in flowering stage against a broad spectrum of bacteria. Micrococcus luteus, Pseudomonas aeruginosa and Candida spp. were the most sensitive to this plant material. Total phenolic content for the methanol extract of L. mutellina herb received from plants in flowering stage was 1.56 g of chlorogenic acid equivalents/100 g dry weight. The methanol extract of L. mutellina herb received from plants in flowering stage showed antioxidant activity with DPPH (IC50 value of 0.40 mg mL(-1)) and with ABTS (IC50 value of 8.65 mg mL(-1)).

Synthesis and antibacterial activity of novel fused 1,3-thiazoles and 1,3-thiazines incorporating a 2,4-dihydroxyphenyl residue.

In an attempt to find a new class of antimicrobial agents, a series of benzothiazoles, 1,3-thiazolo[5,4-b]pyridines, 4H-3,1-benzothiazines, naphtho[2,3-d][1,3]thiazole-4,9-diones and other related compounds containing a 2,4-dihydroxyphenyl moiety were prepared. They were obtained via the reaction of aryl-modified sulfinyl[bis(2,4-dihydroxyphenylmethanethione)]s with appropriate commercial chemical reagents in the endocyclization processes. The MIC values of the compounds towards eight reference bacterial strains were assessed by the two-fold serial micro-dilution broth method. They exhibited inhibitory effects against the Gram-positive strains tested opposite to Gram-negative ones. Some compounds were more effective than the reference drug. 4-(6-Chloro-4H-3,1-benzothiazin-2-yl)-6-methylbenzene-1,3-diol (5b) due to its very good activity (MIC from 1.56 to 3.13 µg/mL) and low cytotoxicity (IC(50) > 50 µg/mL) may be regarded as a promising precursor for the development of novel antibacterial agents.

Anaerobic bacteria colonizing the lower airways in lung cancer patients.

Anaerobes comprise most of the endogenous oropharyngeal microflora, and can cause infections of airways in lung cancer patients who are at high risk for respiratory tract infections. The aim of this study was to determine the frequency and species diversity of anaerobes in specimens from the lower airways of lung cancer patients. Sensitivity of the isolates to conventional antimicrobial agents used in anaerobe therapy was assessed. Respiratory secretions obtained by bronchoscopy from 30 lung cancer patients were cultured onto Wilkins-Chalgren agar in anaerobic conditions at 37°C for 72-96 hours. The isolates were identified using microtest Api 20A. The minimal inhibitory concentrations for penicillin G, amoxicillin/clavulanate, piperacillin/tazobactam, cefoxitin, imipenem, clindamycin, and metronidazole were determined by E-test. A total of 47 isolates of anaerobic bacteria were detected in 22 (73.3%) specimens. More than one species of anaerobe was found in 16 (53.3%) samples. The most frequently isolated were Actinomyces spp. and Peptostreptococcus spp., followed by Eubacterium lentum, Veillonella parvula, Prevotella spp., Bacteroides spp., Lactobacillus jensenii. Among antibiotics used in the study amoxicillin/clavulanate and imipenem were the most active in vitro (0% and 2% resistant strains, respectively). The highest resistance rate was found for penicillin G and metronidazole (36% and 38% resistant strains, respectively). The results obtained confirm the need to conduct analyses of anaerobic microflora colonizing the lower respiratory tract in patients with lung cancer to monitor potential etiologic factors of airways infections, as well as to propose efficient, empirical therapy.

Antimicrobial activity of fatty acids from fruits of Peucedanum cervaria and P. alsaticum.

Plants of the genus Peucedanum have been used in traditional medicine for a long time to treat different diseases including infectious diseases. The hexane fruits extracts of Peucedanum cervaria and P. alsaticum were examined for antimicrobial activity and analyzed for their fatty acid content. Fatty acid composition of oils were analyzed by GC/FID in methyl ester form. Minimal inhibitory concentrations (MICs) of fatty acid fractions against twelve reference bacterial and yeast strains were performed by the twofold serial microdilution broth method. Fourteen fatty acids were identified. Oleic and linoleic acids were found to be dominant. The extracts from both plants examined exhibited inhibitory effects against Gram-positive strains tested with different MIC values (0.25-2 mg/ml); however, extract from P. alsaticum possessed stronger antibacterial properties and a broader spectrum. The growth of Gram-negative bacteria and Candida spp. strains was not inhibited even at the highest extract concentration used (MIC>4 mg/ml). Standard fatty acids exhibited inhibitory effects towards all bacterial and yeast strains used in this study; however, the majority of bacteria were more sensitive to linoleic than to oleic acid. These results revealed, for the first time, that hexane extracts obtained from fruits of P. alsaticum and P. cervaria possess moderate in vitro antibacterial activity against Gram-positive bacteria including staphylococci. Linoleic and oleic acids appear to be the compounds responsible for this effect, and a synergistic antimicrobial effect between these two fatty acids was indicated.

Comparison of hydrodistillation and headspace solid-phase microextraction techniques for antibacterial volatile compounds from the fruits of Seseli libanotis.

The volatile compounds from the fruits of Seseli libanotis, wild-growing in Poland, were obtained by hydrodistillation (HD) and headspace solid-phase microextraction techniques (HS-SPME) and analyzed using GC-MS. A total of 58 components were identified. The most abundant compounds were sabinen and beta-phellandrene. MIC (minimal inhibitory concentration) and MBC (minimal bactericidal concentration) values for the essential oil against reference bacterial strains and clinical S. aureus isolates were determined. The essential oil showed better antibacterial activity against the Gram-positive bacteria (MICs between 0.15 to 1.25 mg/mL) when compared with Gram-negative bacteria (MICs between 1.25 to 2.5 mg/mL). The low values of the MBC/MIC ratio suggest that the examined oil behaved as a bactericidal agent.

A comparative analysis of phenotypic and genotypic methods for the determination of the biofilm-forming abilities of Staphylococcus epidermidis.

The collection of 146 Staphylococcus epidermidis strains isolated from the nasopharynx of lung cancer patients has been studied for the ability of slime secretion and biofilm formation using the Congo red agar (CRA) test and the microtiter plate (MtP) method, respectively. The prevalence of the icaAD and the aap genes was also analyzed. Some isolates (35.6%) were biofilm positive by the MtP method, while 58.9% of isolates exhibited a slime-positive phenotype by the CRA test. The sensitivities of the CRA test evaluated using the MtP method as a gold standard of biofilm production were 73.1%, 97.3% and 13.3% for all the strains screened, ica-positive and ica-negative strains, respectively. The genotype ica(+)aap(+) was correlated with a strong biofilm-producer phenotype. Interestingly, some of the ica(-)aap(-) isolates could also form a biofilm. The correlation between the presence of icaAD genes and the biofilm-positive phenotype by the MtP method as well as slime production by the CRA test was statistically significant (P<0.0001). However, some S. epidermidis strains possess the potential ability of ica-independent biofilm formation; thus, further studies are needed to determine reliable, short-time criteria for an in vitro assessment of biofilm production by staphylococci.

Bacterial colonization of pleural drains in patients with lung cancer: an analysis of risk factors.

BACKGROUND: Microbial colonization of pleural drains during the postoperative period may be regarded as an important factor in infection development. The aim of this paper was to determine the frequency and risk factors associated with microbial colonization of pleural drains in patients with non-small-cell lung cancer (NSCLC) who underwent surgical resection. MATERIAL/METHODS: A total of 72 pleural drain fluids obtained from 36 patients with NSCLC were microbiologically examined. RESULTS: Microbial colonization of pleural drain fluid was noted in 30/36 patients (83.34%). Aerobic and anaerobic bacteria were found in 13 (36.11%) and 7 (19.44%) patients, respectively, and mixed microflora were cultured from 9 (25%). Non-fermentative gram-negative rods and coagulase-negative staphylococci were mainly isolated. Univariate analysis revealed that FEV1% >65 and FEV1/FVC were significantly related to drain colonization by bacteria in general, FEV1/FVC to colonization by aerobic bacteria, and hospitalization >5 days before surgery to colonization by anaerobic bacteria. According to multivariate analysis, application of antibiotic prophylaxis other than cefuroxime or ceftriaxone and FEV1% >65 were the independent factors related to drain colonization by bacteria in general; antibiotic prophylaxis other than cefuroxime or ceftraixone alone and hospitalization >5 days before surgery to colonization by anaerobic bacteria; and a higher rate of FEV1/FVC, no chemotherapy before surgery, and postsurgical complications after surgery to colonization by aerobic bacteria. CONCLUSIONS: Patients with resectable lung cancer showed a high rate of pleural drain colonization, mainly by opportunistic pathogens, even in the absence of clinical signs of infection.

Volatile compounds in fruits of Peucedanum cervaria (Lap.) L.

The volatile compounds from Peucedanum cervaria (Lap.) L. were obtained by hydrodistillation (HD) and headspace solid-phase microextraction techniques (HS-SPME), and then analyzed by GC/MS methods. The composition of samples from a botanical garden was compared with plants collected in the wild. The main compounds of the essential oils of P. cervaria were identified as alpha-pinene, sabinene, and beta-pinene (more than 80% of oil). The content of beta-myrcene, limonene+beta-phellandrene, and germacrene D was higher than 1%. The in vitro antibacterial activity of the essential oil was evaluated by the agar dilution method against ten reference strains of Gram-positive and Gram-negative bacteria.

Comparison of microdilution method and E-test procedure in susceptibility testing of caspofungin against Candida non-albicans species.

It is widely known that systemic and mucosal candidiasis caused by Candida non-albicans strains endangers the lives of hospitalised patients since these pathogens are extremely difficult to defeat by commonly used antifungal agents. The present study determined the in vitro activities of a novel antimicotic drug - caspofungin - against 76 Candida non-albicans isolates by means of the CLSI reference method for broth dilution antifungal susceptibility testing of yeasts and the E-test procedure for comparison. Caspofungin was efficacious against the majority of strains tested, with the average MICs90 evaluated by the microdilution method and E-tests amounting to 1 mg/l and 0.5 mg/l, respectively. Since the agreement between MICs within +/-2 dilutions obtained by these two techniques was 92% (Kappa coefficient of 0.92), the E-test procedure seems to be a reliable alternative to the broth microdilution method and may provide another choice for clinical laboratories.

Dynamics of nasopharyngeal colonization by gram-negative rods in patients with resectable lung cancer during short-term hospitalization.

We determined the frequency of colonization of nasopharynx by Gram-negative rods in 63 patients with lung cancer undergoing thoracic surgery who routinely receive antimicrobial prophylaxis. Throat and nasal specimens were taken from each patient twice: on the day of hospital admission (examination I) followed by thoracic surgery and on the fourth day after thoracic surgery (examination II). The isolated strains were identified using API 20E or API 20NE microtests. Susceptibility to selected antimicrobial agents was detected by the disc diffusion method according to Clinical Laboratory Standards Institute recommendations. A total of 27 strains of Gram-negative rods were cultured from 21 patients. During short-term hospitalization, in patients with lung cancer undergoing thoracic surgery and preoperative prophylaxis, qualitative and quantitative changes in Gram-negative rods colonizing the nasopharynx were observed. A statistically significant increase in the frequency of these bacteria on mucous membranes of nasopharynx in examination II was found (Chi-squared test, p<0.00001). The strains of Enterobacteriaceae were highly susceptible to antimicrobial agents, whereas most of the non-fermenting rods were classified as multi-drug resistant organisms.

Slime production and cell surface hydrophobicity of nasopharyngeal and skin staphylococci isolated from healthy people.

The collection of 314 staphylococcal strains including Staphylococcus aureus and coagulase-negative staphylococci (CNS) was isolated from skin or nasopharynx of healthy people. It was found that the majority of staphylococci possessed the ability to produce slime intensively or moderately, irrespective of ecological niche-nose, throat or skin. Most of them showed the hydrophilic cell surface. However, among S. aureus skin isolates or CNS throat isolates predominated strains with hydrophobic cell surface. There was a slight correlation between slime production and the nature of cell surface among CNS isolates but not among S. aureus strains. It was found that most of slime-producing CNS strains showed hydrophilic cell surface, while slime-negative isolates usually possessed hydrophobic cell surface. Our data suggest that slime production but not cell surface hydrophobicity can be regarded as an essential colonization factor responsible for staphylococci adherence to skin or mucous membranes of upper respiratory tract. These data also suggest that slime production seems to be a general feature of staphylococci isolated from various niches of healthy people.

[Hydrophobic properties of gram-negative rods colonizing upper respiratory tract of healthy people]. / Hydrofobowe wlasciwosci paleczek gram-uje- mnych kolonizujacych górne drogi oddechowe zdrowych osób.

The cell surface hydrophobicity is one of the non specific factors of adhesion influencing the ability of microorganisms to colonize nasopharynx. The aim of this paper was to evaluate via salt aggregation test (SAT) the cell surface hydrophobicity of 150 strains of gram-negative rods isolated from the throat or/and nasal specimens of healthy people. It has been found that among the nonfermenting rods hydrophobic strains were predominant. In contrast, the isolates of Enterobacteriaceae family were characterized by the distinctive features of the cell surface within particular genera or even species. The obtained results show that, despite differences in cell surface hydrophobicity, numerous species of gram-negative rods have the ability to colonize the mucous membrane of upper respiratory tract. This suggests that the cell surface hydrophobicity is rather a feature of species or genus, but it is not related to the ecological niche of microorganisms in human body.

[Phenotypic characteristics of coagulase-negative Staphylococci colonizing pleural drains in patients with lung cancer after thoracic surgery]. / Fenotypowa charakterystyka gronkowców koagulazo-ujemnych kolonizujacych dreny po zabiegach torakochirurgicznych u pacjentów z rakiem pluca.

Coagulase-negative staphylococci (CNS) represent an important group of etiologic agents of infections associated with plastic biomaterials, e.g. drains. In the present paper 33 strains of CNS were characterized. All of them were isolated from fluid of pleural drains in patients with lung cancer after pulmonary resection under conditions of antimicrobial prophylaxis. The most frequently isolated species were Staphylococcus epidermidis and S. warneri. The majority of CNS strains showed ability to produce slime and possessed hydrophobic properties of cell surface. Strains of CNS resistant to penicillin and oxacillin, but sensitive to amoxicillin/clavulanate were isolated most frequently. Only two methicillin-resistant strains, belonging to S. haemolyticus, were found. The obtained data indicate that CNS strains colonizing pleural drains had potential ability to adhere to smooth surfaces. Most of isolated strains were susceptible to antibiotics and chemotherapeutics routinely used in staphylococcal infections.