Microarray Analysis of the Genomic Effect of Eugenol on Methicillin-Resistant Staphylococcus aureus.

Staphylococcus aureus is a highly adaptive human pathogen responsible for serious hospital- and community-acquired infectious diseases, ranging from skin and soft tissue infections, to complicated and life-threatening conditions such as endocarditis and toxic shock syndrome (TSS). The rapid development of resistance of this organism to available antibiotics over the last few decades has necessitated a constant search for more efficacious antibacterial agents. Eugenol (4-allyl-2-methoxyphenol) belongs to the class of chemical compounds called phenylpropanoids. It is a pure-to-pale yellow, oily liquid substance, mostly extracted as an essential oil from natural products such as clove, cinnamon, nutmeg, basil, and bay leaf. Eugenol has previously been shown to have antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA). However, the mechanism of action of eugenol against MRSA has not, as yet, been elucidated; hence, the necessity of this study. Global gene expression patterns in response to challenge from subinhibitory concentrations of eugenol were analysed using the Agilent DNA microarray system to identify genes that can be used as drug targets-most importantly, essential genes involved in unique metabolic pathways elicited for bacterial survival. Transcriptomic analysis of fluctuating genes revealed those involved in amino acid metabolism, fatty acid metabolism, translational, and ribosomal pathways. In amino acid metabolism, for instance, the argC gene encodes for N-acetyl-gamma-glutamyl-phosphate reductase. The argC gene plays an important role in the biosynthesis of arginine from glutamate in the amino acid metabolic pathway. It is the enzyme that catalyses the third step in the latter reaction, and without this process the production of N-acetylglutamate 5-semialdehyde cannot be completed from the NADP-dependent reduction of N-acetyl-5-glutamyl phosphate, which is essential for the survival of some microorganisms and plants. This study enables us to examine complete global transcriptomic responses in MRSA when challenged with eugenol. It reveals novel information with the potential to further benefit the exploratory quest for novel targets against this pathogen, with a view to the development of efficacious antimicrobial agents for the treatment of associated infections.

In vitro methods used for discovering plant derived products as wound healing agents - An update on the cell types and rationale.

Wounds still pose a huge burden on human health and healthcare systems in many parts of the world. Phytomedicines are being used to heal the wounds since ancient times. Now-a-days also many researchers are exploring the wound healing activity of phytomedicines. Wound healing is a complex process thus, it is always a question mark regarding the best test model (in vivo, ex vivo and in vitro) model to assess the wound healing activity of phytomedicines. In general, the researchers would opt for in vivo model - probably because of closer physiological relevance to human wounds. However, in vivo experimental models are not suitable for high throughput screening and not ethical in terms of initial screening of the phytomedicines. The in vivo models are associated with difficulties in obtaining the ethical approvals, requires huge budget, and resources. We argue that judicious selection of cell types would serve the purpose of developing a physiologically relevant in vitro experimental model. A lot of progress has been made in molecular biology techniques to bridge the gap between in vitro models and their physiological relevance. The in vitro models are the best suited for high throughput screening and to elucidate the molecular mechanisms. The main aim of this review is to provide insights on selection of the cell types for developing physiologically relevant in vitro wound healing assays, which can be used to improve the value of phytomedicines further.

Multidrug resistant yeasts in synanthropic wild birds.

BACKGROUND: The aim of this study was to investigate the presence of multidrug resistant yeasts in the faeces of synanthropic wild birds from the Bangsar suburb of Kuala Lumpur. METHODS: Species characterisations of yeast isolates and determinations of antimycotic susceptibility profiles were undertaken using the commercial characterization kit, Integral System Yeasts Plus (Liofilchem, Italy). RESULTS: Fourteen species of yeasts were detected in the bird faecal samples.Candida albicans was present in 28.89% of bird faecal samples, Candida krusei (13.33%), Candida tropicalis (4.44%), Candida glabrata (4.44%), Candida parapsilosis (2.22%), Candida lambica (2.22%), Candida stellatoidea (2.22%), Candida rugosa (2.22%) and Candida lusitaniae (2.22%). Amongst the non-candidal yeast isolates, Cryptococcus laurentii was present in 6.67% of bird faecal samples, Cryptococcus uniguttulatus (4.44%), Saccharomyces cerevisiae (4.44%), Trichosporon pullulans (2.22%), Trichosporon pullulans/Cryptococcus albidus (8.89%) and Rhodotorula rubra/Rhodotorula glutinis (4.44%). Of the isolated yeasts, 18.1% (or 26/144) were found to be resistant to all 11 antimycotic agents they were tested against i.e. Nystatin, Amphotericin B, Flucytosine, Econazole, Ketoconazole, Clotrimazole, Miconazole, Itraconazole, Voriconazole, Fluconazole 16 and Fluconazole 64. 45.8% (or 66/144) of the bird faecal yeast isolates were resistant to four or more of the 11 antimycotic agents they were tested against. CONCLUSIONS: This finding is of public health significance as these synanthropic wild birds may be reservoirs for transmission of drug resistant yeast infections to humans.

In vitro antibacterial effects of Cinnamomum extracts on common bacteria found in wound infections with emphasis on methicillin-resistant Staphylococcus aureus.

ETHNOPHARMACOLOGICAL RELEVANCE: Cinnamomum species have been widely used in many traditional systems of medicine around the world. In the Malaysian traditional system of medicine, the leaves, stem bark and stem wood of Cinnamomum iners, Cinnamomum porrectum, Cinnamomum altissimum and Cinnamomum impressicostatum have been used to treat wound infections. To study the antibacterial effects of Cinnamomum iners, Cinnamomum porrectum, Cinnamomum altissimum and Cinnamomum impressicostatum against common bacteria found in wound infections with primary focus on methicillin-resistant Staphylococcus aureus (MRSA). MATERIALS AND METHODS: The crude extracts from the leaves, stem-bark and stem-wood of Cinnamomum iners, Cinnamomum porrectum, Cinnamomum altissimum and Cinnamomum impressicostatum were obtained using sequential extraction with hexane, ethylacetate, methanol and water. The volatile oils were obtained by hydro-distillation. The antibacterial activities of extracts were investigated using disk diffusion assays and broth microdilution assays. RESULTS: The volatile oils obtained from the stem-bark of Cinnamomum altissimum, Cinnamomum porrectum and Cinnamomum impressicostatum have shown significant antibacterial activity against a wide range of Gram positive and Gram negative bacteria including MRSA. A few test extracts have shown better activity against MRSA as compared to methicillin sensitive Staphylococcus aureus (MSSA). Amongst all the test extracts, Cinnamomum impressicostatum stem-bark water extract produced the largest inhibition zone of 21.0mm against MRSA while its inhibition zone against MSSA was only 8.5mm. The minimum inhibitory concentration (MIC) of this extract against MRSA was 19.5 µg mL(-1) and the corresponding minimum bactericidal concentration (MBC) was 39.0 µg mL(-1). CONCLUSIONS: This study has scientifically validated the traditional use of Cinnamomum species in treating wound infections. Of high scientific interest was the observation that the antibacterial effect of Cinnamomum impressicostatum stem-bark crude water extract against MRSA was significantly higher than its effect against MSSA, suggesting that the extract contains a compound(s) with higher specific neutralising activity against the drug resistance markers of MRSA.

Bacterial constituents of indoor air in a high throughput building in the tropics.

Airborne bacteria are significant biotic constituents of bioaerosol. Bacteria at high concentrations in the air can compromise indoor air quality (IAQ) and result in many diseases. In tropical environments like Malaysia that extensively utilize air-conditioning systems, this is particularly significant due to continuous recirculation of indoor air and the potential implications for human health. Currently, there is a lack of knowledge regarding the impact of airborne bacteria on IAQ in Malaysia. This study was prompted by a need for reliable baseline data on airborne bacteria in the indoor environment of tropical equatorial Malaysia, that may be used as a reference for further investigations on the potential role played by airborne bacteria as an agent of disease in this region. It was further necessitated due to the threat of bioterrorism with the potentiality of release of exotic pathogenic microorganisms into indoor or outdoor air. Before scientists can detect the latter, a gauge of the common microorganisms in indoor (as well as outdoor) air needs to be ascertained, hence the expediency of this study. Bacterial counts from the broad-based and targeted study were generally in the order of 10(2) colony-forming units (CFU) per m(3) of air. The most prevalent airborne bacteria found in the broad-based study that encompassed all five levels of the building were Gram-positive cocci (67.73%), followed by Gram-positive rods (24.26%) and Gram-negative rods (7.10%). Gram-negative cocci were rarely detected (0.71%). Amongst the genera identified, Kytococcus sp., Micrococcus sp., Staphylococcus sp., Leifsonia sp., Bacillus sp. and Corynebacterium sp. predominated in indoor air. The most dominant bacterial species were Kytococcus sedentarius, Staphylococcus epidermidis and Micrococcus luteus. The opportunistic and nosocomial pathogen, Stenotrophomonas maltophilia was also discovered at a high percentage in the cafeteria. The bacteria isolated in this study have been increasingly documented to cause opportunistic infections in immuno-compromised patients, sometimes with fatal outcomes. Furthermore, some of them are becoming increasingly resistant to antibiotics. Hence, we propose that indoor reservoirs of these bacteria and their associated clinical and more subtle health effects, if any, be investigated further.