Carvacrol/clay hybrids loaded into in situ gelling films.

The aim of the present work was the development of polymer films loaded with a carvacrol (CVR)/clay hybrid (HYBD) for the delivery of CRV in infected skin ulcer treatment. Different clays were considered: montmorrilonite, halloysite and palygorskite (PHC). CRV incorporation in PHC reduced its volatility. HYBD showed 20% w/w CRV loading capacity and was able to preserve CRV antioxidant properties. HYBD was characterized by improved antimicrobial properties against S. aureus and E. coli and cytocompatibility towards human fibroblasts with respect to pure CRV. Films were prepared by casting an aqueous dispersion containing poly(vinylalcohol) (PVA), poly(vinylpyrrolidone) (PVP), chitosan glutamate (CS), sericin and HYBD. Optimization of film composition was supported by a Design of Experiments (DoE) approach. In a screening phase, a full factorial design (FFD) was used and the following factors were investigated at two levels: PVA (12-14%w/w), PVP (2-4%w/w) and CS (0.134-0.5%w/w) concentrations. For the optimization phase, FFD was expanded to a "central composite design". The response variables considered were: elongation, tensile strength and buffer absorption of films, durability of the gels formed after film hydration. Upon hydration, the optimized film formed a viscoelastic gel able to protect the lesion area and to modulate CRV release.

In vitro evaluation of the wound healing activity of Drypetes klainei stem bark extracts.

ETHNOPHARMACOLOGICAL RELEVANCE: Drypetes klainei Pierre ex Pax is used in Cameroon by Baka Pygmies in the wound healing process and for the treatment of burns. AIM OF THE STUDY: To validate the traditional use of D. klainei Pierre ex Pax stem bark extracts through the evaluation of their antimicrobial properties and their ability to improve wound healing process in fibroblast cell cultures. MATERIALS AND METHODS: The antimicrobial properties of D. klainei extracts were evaluated against Staphylococcus aureus ATCC 6538, Streptococcus pyogenes ATCC 19615, Escherichia coli ATCC 10536, Candida albicans ATCC 10231, on the basis of the minimum inhibitory concentration (MIC) and the minimum bactericidal-fungicidal concentration (MBC-MFC) by the macrodilution method. The extracts abilities to accelerate wound healing were studied on murine and human fibroblasts in terms of cell viability and migration (scratch wound-healing assay). RESULTS: All the extracts were non-toxic against the selected microorganisms at the tested concentrations, and significantly improve wound healing process in vitro, compared to untreated controls. However, the defatted methanol extract was active at lower concentrations, compared to the water extract. CONCLUSIONS: The ability of both water and defatted methanol extracts to accelerate scratch wound closure in fibroblast cultures may support the traditional use of D. klainei stem bark in the treatment of skin lesions (such as burns) even if no antimicrobial activity was evidenced.

Anti-inflammatory, antimicrobial and antioxidant activities of Diospyros bipindensis (Gürke) extracts and its main constituents.

ETHNOPHARMACOLOGICAL RELEVANCE: Diospyros bipindensis (Gürke) stem bark is used in Cameroon by Baka Pygmies for the treatment of respiratory disorders. AIM OF THE STUDY: To assess the anti-inflammatory, antibacterial and antioxidant properties of constituents from the bark extracts through bioassay-guided fractionation. MATERIALS AND METHODS: The anti-inflammatory activity of extracts, fractions and pure compounds was assessed through the inhibition of the pro-inflammatory mediator nuclear factor-kappa B (NF-κB) transcriptional activity and nitric oxide (NO) production. DPPH, ABTS and ORAC assays were used for determining the antioxidant properties. The activity against Streptococcus pneumoniae, Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli and Klebsiella pneumoniae, was evaluated on the basis of the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) by the macrodilution method. RESULTS: The water extract showed antimicrobial activity against S. pneumoniae (MIC: 300 µg/ml) and S. pyogenes (MIC: 300 µg/ml). The dichloromethane extract efficiently inhibited NF-κB transcriptional activity and NO production and exhibited significant antioxidant activity in the ORAC assay. An interesting activity was also found against S. pneumoniae (MIC: 200 µg/ml), S. aureus (MIC: 400 µg/ml) and S. pyogenes (MIC: 200 µg/ml). The phytochemical investigation of the dichloromethane extract afforded plumbagin, canaliculatin, ismailin, betulinic acid and 4-hydroxy-5-methyl-coumarin as the main constituents. Plumbagin and ismailin were found to be responsible for the main biological activities observed. CONCLUSIONS: These results may provide a rational support for the traditional use of Diospyros bipindensis stem bark in the treatment of respiratory disorders, since the anti-inflammatory, antimicrobial and antioxidant compounds isolated from the dichloromethane extract were also present in the traditional water extract.

Chemical composition and antimicrobial activity of Phyllanthus muellerianus (Kuntze) Excel essential oil.

ETHNOPHARMACOLOGICAL RELEVANCE: Phyllanthus muellerianus (Kuntze) Excel (family Euphorbiaceae) stem bark is used in Cameroon by Baka pygmies as a remedy for wound healing and tetanus. AIM OF THE STUDY: To characterize the chemical composition and to evaluate the antimicrobial properties of the essential oil of the plant. MATERIALS AND METHODS: The essential oil was extracted from the stem bark by dynamic head space and by hydrodistillation and characterized by GC and GC-MS analyses. The antimicrobial activity was evaluated on the basis of the minimum inhibitory concentration (MIC) and the minimum bactericidal-fungicidal concentration (MBC-MFC) by the micro and macrodilution methods. The following bacteria and fungi were used: Clostridium sporogenes ATCC 3584, Staphylococcus aureus ATCC 6538, Streptococcus mutans ATCC 25175, Streptococcus pyogenes ATCC 19615, Escherichia coli ATCC 10536, Candida albicans ATCC 10231, Candida albicans LM 450, Trichophyton mentagrophytes LM 230, Trichophyton rubrum LM 237, Microsporum canis LM 324. RESULTS: The hydrodistillation afforded 0.06% (dry weight basis) of pale yellow oil. Thirty-eight compounds representing 90.69% were identified. The major component (36.40%) was found to be (E)-isoelemicin, identified by comparison of its (1)H-NMR experimental data, with literature data. The oil showed good antibacterial activity against Clostridium Sporogenes, Streptococcus mutans and Streptococcus pyogenes with MIC ranging from 13.5 to 126 µg/ml. A weak antifungal activity (MIC 250 µg/ml) was found against Trichophyton rubrum, only. CONCLUSIONS: The antimicrobial activity and the chemical composition of Phyllanthus muellerianus stem bark essential oil are reported for the first time.

Antimicrobial properties of stem bark extracts from Phyllanthus muellerianus (Kuntze) Excell.

ETHNOPHARMACOLOGICAL RELEVANCE: The plants of the genus Phyllanthus (Euphorbiaceae) are widely distributed in most tropical and subtropical countries, and have long been used in folk medicine to treat several diseases. Particularly, Phyllanthus muellerianus (Kuntze) Excell, commonly called "mbolongo" in Cameroon, is used by pygmies baka as a remedy for tetanus and wound infections. AIM OF THE STUDY: To investigate the antimicrobial properties of Phyllanthus muellerianus (Kuntze) Excell (family Euphorbiaceae) stem bark used in Cameroon by baka pygmies as a remedy for wound healing and tetanus. MATERIALS AND METHODS: Aqueous and methanol extracts with and without defatting treatment, were prepared and their activity against Clostridium sporogenes ATCC 3584, Staphylococcus aureus ATCC 6538, Streptococcus mutans ATCC 25175, Streptococcus pyogenes ATCC 19615, Escherichia coli ATCC 10536, Candida albicans ATCC 10231, was evaluated on the basis of the minimum inhibitory concentration (MIC) and the minimum bactericidal-fungicidal concentration (MBC-MFC) by the macrodilution method. RESULTS: Water extract showed a weak activity against Clostridium sporogenes (MIC 900 µg/mL) and resulted inactive at the tested concentrations against all the other microorganisms. The defatted methanol extract, inactive against Staphylococcus aureus, Escherichia coli, Candida albicans, exhibited a very interesting activity against Clostridium sporogenes and Streptococcus pyogenes (MIC 100 µg/mL and 300 µg/mL, respectively), which seems to validate the use of this plant in pygmies traditional medicine for the treatment of tetanus and wound infections. The activity found against Streptococcus mutans (300 µg/mL), aetiological agent of caries, may suggest a possible use of this plant as natural remedy to prevent dental diseases. CONCLUSIONS: The activity against streptococci and Clostridium sporogenes ATCC 3584, showed by stem bark extracts of Phyllanthus muellerianus, traditionally used by baka pygmies to treat wound infections and tetanus, is reported for the first time.

Pyrogenic activity of air to characterize bioaerosol exposure in public buildings: a pilot study.

AIMS: The aim of our study was to investigate indoor air quality (IAQ) by comparing pyrogen concentration and microbiological contamination in offices in public buildings. METHODS AND RESULTS: Air samples were collected during cold and warm seasons in 39 offices in four European cities. Pyrogens were measured by the in vitro pyrogen test (IPT), moulds and bacteria by classical microbiology. In 92% of the investigated offices, pyrogen and microbial contaminations were below 150 EEU m(-3) and 10(3) CFU m(-3), respectively, whilst in 75%, moulds did not exceed 10(2) CFU m(-3). CONCLUSIONS: The IPT is a rapid, reliable tool for measuring pyrogens that could be used as an 'early warning' indicator of IAQ. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study on pyrogenic compound detection in offices using IPT, which could serve for developing future indoor air guidelines.

Thermally sensitive gels based on chitosan derivatives for the treatment of oral mucositis.

The aim of the present work was the development of a thermally sensitive mucoadhesive gel based on chitosan derivatives for the treatment of oral mucositis. Trimethyl chitosan (TMC) and methylpyrrolidinone chitosan (MPC) were considered. They were mixed with glycerophosphate (GP) according to different polymer/GP molar ratios and characterized for gelation properties by means of rheological analysis in comparison with chitosan. The influence of molecular weight and substitution degree (SD) of TMC on gelation temperature and time was investigated. The mucoadhesive properties of the mixtures were also assessed using porcine buccal mucosa. The best properties were shown by TMC with high MW and low SD mixed with GP according to 1:2molar ratio. Such mixture was loaded with benzydamine hydrochloride, an anti-inflammatory drug with antimicrobial properties and subjected to in vitro drug release and wash away test. The formulation based on TMC/GP mixture was able to prolong drug release and to withstand the removal physiological mechanisms. The antimicrobial properties of both vehicle and formulation were investigated. Also in absence of drug, TMC/GP mixture was characterized by antimicrobial properties.

Validation of use of a traditional remedy from Bridelia grandis (Pierre ex Hutch) stem bark against oral Streptococci.

AIM OF THE STUDY: Bridelia grandis (Pierre ex Hutch) (Euphorbiaceae) is a medicinal plant traditionally used in Cameroon by pygmies Baka as a remedy for oral cavity affection. Bioassay-guided stem bark extracts were investigated for their in vitro antimicrobial properties as well as their phytochemical constituents. MATERIALS AND METHODS: The first extraction was carried out according to the traditional use. Further extractions were carried out with solvents of different polarity such as methanol (MeOH), ethyl acetate (EtOAc) and mixtures of MeOH-H2O. The antimicrobial activity of the extracts against oral Streptococci was evaluated on the basis of the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) by the macrodilution method; the bacterial surface hydrophobicity was also evaluated. RESULTS: Water, methanol and mixtures methanol-water extracts, exhibited antibacterial activity with MIC between 0.5 and 2mg/ml justifying the traditional use of Bridelia grandis stem bark for oral cavity affection. Preliminary phytochemical analysis was performed on the most active extract (methanol) using appropriate tests and well established analytical screening methods, such as TLC and RP-HPLC/DAD. CONCLUSIONS: The data obtained indicate that tannins constitute the chemical family responsible for the biological activity.

Micro-organisms and dust exposure in an Italian grain mill.

AIMS: In order to assess possible occupational risk for workers in a grain mill, we evaluated aerial microbiological contamination in different areas of the mill and at different points of the production line. We also measured the concentration of aerodispersed dust particles. METHODS AND RESULTS: An assessment of microbiological contamination levels based on a Global Index of Microbial Contamination per cubic metre (GIMC per m3), an Index of Mesophilic Bacterial Contamination, and an Amplification Index is proposed. The indices were obtained from total and fungal counts. The cleaning sector is the most contaminated area of the mill: the mean GIMC per m3 was 17,213.6. In this area, the average microbial contamination was 11.41 times higher than that in the external environment. The highest concentrations of aerodispersed dust (inhalable 2.763 mg m(-3); respirable 1.400 mg m(-3)) were found in the cleaning area. CONCLUSIONS: The proposed microbiological indices and the concentrations of aerodispersed dust particles show that the most hazardous section of the mill is the cleaning area. The large variation in the data does not depend on seasonal factors, but rather on not easily identifiable conditions of the internal environment which facilitate diffusion and/or proliferation of the micro-organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: The proposed microbiological contamination indices and the evaluation of the concentration of dust particles allow the identification of critical positions during the production cycle so that suitable measures to prevent the aerial contamination can be taken.

Determination of aerial microbiological contamination in scholastic sports environments.

AIMS: To assess the microbiological indoor air quality (IAQ) of high school and college gyms during physical training lessons and to evaluate the effective microbiological exposure of students. METHODS AND RESULTS: Air samples from 11 high school and college gyms of Pavia, Italy were collected. Total bacterial counts, total fungal counts and characterization of fungal taxa were determined. Air quality was evaluated using three microbiological contamination indices: the global index of microbiological contamination per m3 (GIMC per m3), the index of mesophilic bacterial contamination (IMC) and the amplification index (AI). CONCLUSIONS: This work testifies that air contamination in indoor gyms is always superior to that of the outdoor environment. Nevertheless, students are exposed to relatively low concentrations of airborne micro-organisms. The highest values of fungal counts and GIMC per m3 (>14,661) were observed between April and October when the central heating systems were switched off. The lowest fungal counts were detected in modern buildings equipped with forced ventilation systems. From qualitative aeromycological studies, 45 fungal taxa were identified, and different potentially allergenic species were isolated. SIGNIFICANCE AND IMPACT OF THE STUDY: The standardization of air sampling methods and the correct evaluation of aeromicrobiological results allow the classification of indoor air healthiness. The proposed microbiological contamination indices together with the characterization of airborne fungal taxa are useful tools for detailed description of IAQ.

Antiadhesive effect of green and roasted coffee on Streptococcus mutans' adhesive properties on saliva-coated hydroxyapatite beads.

Green and roasted coffees of the two most used species, Coffea arabica and Coffea robusta, several commercial coffee samples, and known coffee components were analyzed for their ability to interfere with Streptococcus mutans' sucrose-independent adsorption to saliva-coated hydroxyapatite (HA) beads. All coffee solutions showed high antiadhesive properties. The inhibition of S. mutans' adsorption to HA beads was observed both when coffee was present in the adsorption mixture and when it was used to pretreat the beads, suggesting that coffee active molecules may adsorb to a host surface, preventing the tooth receptor from interacting with any bacterial adhesions. Among the known tested coffee components, trigonelline and nicotinic and chlorogenic acids have been shown to be very active. Dialysis separation of roasted coffee components also showed that a coffee component fraction with 1000 Da < MW < 3500 Da, commonly considered as low MW coffee melanoidins, may sensibly contribute to the roasted coffee's antiadhesive properties. The obtained results showed that all coffee solutions have antiadhesive properties, which are due to both naturally occurring and roasting-induced molecules.

[Proposed microbiological indexes for the assessment of air quality in buildings]. / Proposta di indici microbiologici per la valutazione della qualità dell'aria degli edifici.

An assessment of biological risk in the workplace requires the measurement of microorganisms present in the air. Although it is currently difficult to establish a dose-response relationship on the basis of existing epidemiological data, the number and types of microorganisms present offer a useful index when evaluating the quality of the air. Using the results obtained from a study of 226 different offices located in buildings equipped with forced ventilation we propose an assessment of contamination levels based on a Global Index of Microbial Contamination/m3 (GIMC/m3). This index is calculated as the sum of the values of the total microbial count determined for mesophilic bacteria, psychrophilic bacteria and fungi in all sampled areas. GIMC/m3 fluctuates between a minimum value of 23 and a maximum value of 18.069: the mean value varies depending on the ventilation system mode setting, with a value of 145 when operating in the heating mode, 265 in the air-conditioning mode and 258 under simple forced ventilation. 95.5% of the offices under study had a GIMC/m3 value of below 1000: this value is proposed as a threshold limit above which a more extensive assessment of contamination levels based on the measurement of further microbiological indexes should be carried out. To this aim we propose an Index of Mesophilic bacterial Contamination (IMC), obtained by calculating the ratio between the CFU/m3 value measured for mesophilic and psychrophilic bacteria in the same sampling point. The measurement of the Amplification Index (AI) completes the evaluation and this is determined by calculating the ratio between the GICM/m3 values measured inside the building and those measured outside. When considering these observations as a whole we are able to classify the various work areas under study into groups and categories according to the observed values of the proposed microbial contamination indexes.