METHODS: The release of prostaglandin E2 (PGE2) and pro-inflammatory cytokines, tumor necrosis factor (TNF)-α and interleukin (IL)-1β in a culture supernatant was determined by ELISA. Determination of cyclooxygenase-2 (COX-2) protein and the activation of MAPKs molecules (JNK, ERK and p38 MAPK), NF-κB and Akt in LPS-induced U937 human macrophages were investigated by immunoblot technique. The relative gene expression levels of COX-2 and pro-inflammatory cytokines were measured by using qRT-PCR. The major metabolites of P. amarus were qualitatively and quantitatively analyzed in the extract by using validated reversed-phase high performance liquid chromatography (HPLC) methods.

RESULTS: P. amarus extract significantly inhibited the production of pro-inflammatory mediators (TNF-α, IL-1β, PGE2) and COX-2 protein expression in LPS-induced U937 human macrophages. P. amarus-pretreatment also significantly downregulated the increased mRNA transcription of pro-inflammatory markers (TNF-α, IL-1β, and COX-2) in respective LPS-induced U937 macrophages. It downregulated the phosphorylation of NF-κB (p65), IκBα, and IKKα/β and restored the degradation of IκBα, and attenuated the expression of Akt, JNK, ERK, and p38 MAPKs phosphorylation in a dose-dependent manner. P. amarus extract also downregulated the expression of upstream signaling molecules, TLR4 and MyD88, which play major role in activation of NF-κB, MAPK and PI3K-Akt signaling pathways. The quantitative amounts of lignans, phyllanthin, hypophyllahtin and niranthin, and polyphenols, gallic acid, geraniin, corilagin, and ellagic acid in the extract were determined by HPLC analysis.

METHOD: This study proposes a combination of decision tree and logistic regression techniques to model crash severity (injury vs. noninjury), because the combined approach allows the specification of nonlinearities and interactions in addition to main effects. Both a scobit model and a random parameters logit model, respectively accounting for an imbalance response variable and unobserved heterogeneities, are tested and compared. The study data set contains a total of 5 years of crash data (2008-2012) on selected mountainous highways in Malaysia. To enrich the data quality, an extensive field survey was conducted to collect detailed information on horizontal alignment, longitudinal grades, cross-section elements, and roadside features. In addition, weather condition data from the meteorology department were merged using the time stamp and proximity measures in AutoCAD-Geolocation.

RESULTS: The random parameters logit model is found to outperform both the standard logit and scobit models, suggesting the importance of accounting for unobserved heterogeneity in crash severity models. Results suggest that proportion of segment lengths with simple curves, presence of horizontal curves along steep gradients, highway segments with unsealed shoulders, and highway segments with cliffs along both sides are positively associated with injury-producing crashes along rural mountainous highways. Interestingly, crashes during rainy conditions are associated with crashes that are less likely to involve injury. It is also found that the likelihood of injury-producing crashes decreases for rear-end collisions but increases for head-on collisions and crashes involving heavy vehicles. A higher order interaction suggests that single-vehicle crashes involving light and medium-sized vehicles are less severe along straight sections compared to road sections with horizontal curves. One the other hand, crash severity is higher when heavy vehicles are involved in crashes as single vehicles traveling along straight segments of rural mountainous highways.

AIM OF THE STUDY: To test extracts of P. glaucus in a number of bioassays and determine the legitimacy of its traditional use.

MATERIALS AND METHODS: The stems, leaves, roots and fruits of P. glaucus were collected and extracted sequentially with hexane, chloroform and ethanol, respectively. The anti-inflammatory activity was assessed by testing the ability of the extracts to inhibit heat induced protein denaturation, stabilise human red blood cells under hypotonic stress and by testing the inhibitory activity of the extracts against cyclooxygenases 1 and 2. Cytotoxicity was tested using the human lung epithelial cell line MRC-5 and nasopharangeal carcinoma cell line HK1 in the MTT assay.

RESULTS: Many of the samples showed an ability to prevent heat induced protein denaturation, as well as prevent lysis of red blood cells. Most of the extracts demonstrated inhibitory activity towards both of the COX enzymes. The ethanol extracts tended to demonstrate greater toxicity than other extracts, with some of the other extracts significantly enhancing growth and metabolism of the cells.