Genomic DNA extraction from medicinal plants available in Malaysia using a TriOmic(TM) improved extraction kit

DNA extraction was carried out on 32 medicinal plant samples available in Malaysia using the TriOmic(TM) extraction kit. Amounts of 0.1 g flowers or young leaves were ground with liquid nitrogen, lysed at 65°C in RY1(plus) buffer and followed by RNAse treatment. Then, RY2 buffer was added to the samples and mixed completely by vortexing before removal of cell debris by centrifugation. Supernatants were transferred to fresh microcentrifuge tubes and 0.1 volume RY3 buffer was added to each of the transferred supernatant. The mixtures were applied to spin columns followed by a centrifugation step to remove buffers and other residues. Washing step was carried out twice by applying 70% ethanol to the spin columns. Genomic DNA of the samples was recovered by applying 50 μL TE buffer to the membrane of each spin column, followed by a centrifugation step at room temperature. A modification of the TriOmic(TM) extraction procedure was carried out by adding chloroform:isoamyl alcohol (24:1) steps in the extraction procedure. The genomic DNA extracted from most of the 32 samples showed an increase of total yield when chloroform:isoamyl alcohol (24:1) steps were applied in the TriOmicTM extraction procedure. This preliminary study is very important for molecular studies of medicinal plants available in Malaysia since the DNA extraction can be completed in a shorter period of time (within 1 h) compared to manual extraction, which entails applying phenol, chloroform and ethanol precipitation, and requires 1-2 days to complete.     

Oil extraction from Scenedesmus obliquus using a continuous microwave system--design, optimization, and quality characterization

A 1.2 kW, 2450 MHz resonant continuous microwave processing system was designed and optimized for oil extraction from green algae (Scenedesmus obliquus). Algae-water suspension (1:1 w/w) was heated to 80 and 95°C, and subjected to extraction for up to 30 min. Maximum oil yield was achieved at 95°C and 30 min. The microwave system extracted 76-77% of total recoverable oil at 20-30 min and 95°C, compared to only 43-47% for water bath control. Extraction time and temperature had significant influence (p<0.0001) on extraction yield. Oil analysis indicated that microwaves extracted oil containing higher percentages of unsaturated and essential fatty acids (indicating higher quality). This study validates for the first time the efficiency of a continuous microwave system for extraction of lipids from algae. Higher oil yields, faster extraction rates and superior oil quality demonstrate this system's feasibility for oil extraction from a variety of feedstock.     

There Is No Joy like Malicious Joy: Schadenfreude in Young Children

Human emotions are strongly shaped by the tendency to compare the relative state of oneself to others. Although social comparison based emotions such as jealousy and schadenfreude (pleasure in the other misfortune) are important social emotions, little is known about their developmental origins. To examine if schadenfreude develops as a response to inequity aversion, we assessed the reactions of children to the termination of unequal and equal triadic situations. We demonstrate that children as early as 24 months show signs of schadenfreude following the termination of an unequal situation. Although both conditions involved the same amount of gains, the children displayed greater positive expressions following the disruption of the unequal as compared to the equal condition, indicating that inequity aversion can be observed earlier than reported before. These results support an early evolutionary origin of inequity aversion and indicate that schadenfreude has evolved as a response to unfairness.     

Glass fibre reinforced acrylic resin complete dentures: a 5-year clinical study

doi: 10.1111/j.1741‐2358.2010.00385.x Glass fibre reinforced acrylic resin complete dentures: a 5‐year clinical study Background and objectives: The aim of the study was to establish the wear resistance of the glass fiber reinforced complete dentures comparative to the traditional acrylic complete dentures. Materials and methods: Complete new dentures were made to replace old fractured ‘un’‐reinforced acrylic dentures. The total number of dentures was 30 and woven E‐glass fibre reinforcements were used in maxillary complete dentures. Unidirectional E‐glass fibre reinforcements were used as partial fibre reinforcements in mandibular complete dentures. Ten complete acrylic un‐reinforced dentures were used as control. The follow‐up period was 5 years and the recalls were made at 6 months. Results: After 5 years of wearing the new dentures, the control dentures suffered seven fractures. After 5 years all the mandibular reinforced dentures were in good shape. The maxillary complete reinforced dentures suffered four partial fractures. Fracture lines were restricted by the glass fibre net and the patients could still use their dentures. Conclusion: Pre‐impregnated E‐glass fibre nets and polymer pre‐impregnated E‐glass unidirectional fibres are useful in reinforcing acrylic resin complete dentures especially were heavy occlusal forces are involved. Glass fibre reinforcement will be applied on the tension side in both cases (total fibre reinforcement and partial fibre reinforcement). The reinforcement cannot replace the necessary linings and occlusal adjustments.     

A Human Blood-Brain Barrier Transcytosis Assay Reveals Antibody Transcytosis Influenced by pH-Dependent Receptor Binding

We have adapted an in vitro model of the human blood-brain barrier, the immortalized human cerebral microvascular endothelial cells (hCMEC/D3), to quantitatively measure protein transcytosis. After validating the receptor-mediated transport using transferrin, the system was used to measure transcytosis rates of antibodies directed against potential brain shuttle receptors. While an antibody to the insulin-like growth factor 1 receptor (IGF1R) was exclusively recycled to the apical compartment, the fate of antibodies to the transferrin receptor (TfR) was determined by their relative affinities at extracellular and endosomal pH. An antibody with reduced affinity at pH5.5 showed significant transcytosis, while pH-independent antibodies of comparable affinities at pH 7.4 remained associated with intracellular vesicular compartments and were finally targeted for degradation.     

DEF Pocket in p38α Facilitates Substrate Selectivity and Mediates Autophosphorylation

Signaling processes are primarily promoted by molecular recognition and corresponding protein-protein interactions. One of the key eukaryotic signaling pathways is the MAP kinase cascade involved in vital cellular processes such as cell proliferation, differentiation, apoptosis, and stress response. The principle recognition site of MAP kinases, the common docking (CD) region, forms selective interactions with substrates, upstream activators, and phosphatases. A second docking site, defined as the DEF site interaction pocket (DEF pocket), is formed subsequent to ERK2 and p38α activation. Both crystal structures of p38α in its dually phosphorylated form and of intrinsically active mutants showed the DEF pocket, giving motivation for studying its role in substrate activation and selectivity. Mutating selected DEF pocket residues significantly decreased the phosphorylation levels of three p38α substrates (ATFII, Elk-1, and MBP) with no apparent effect on the phosphorylation of MK2 kinase. Conversely, mutating the CD region gave the opposite effect, suggesting p38α substrates can be classified into DEF-dependent and DEF-independent substrates. In addition, mutating DEF pocket residues decreased the autophosphorylation capability of intrinsically active p38α mutants, suggesting DEF-mediated trans-autophosphorylation in p38α. These results could contribute to understanding substrate selectivity of p38α and serve as a platform for designing p38α-selective DEF site blockers, which partially inhibit p38α binding DEF-dependent substrates, whereas maintaining its other functions intact. In this context, preliminary results using synthetic peptides reveal significant inhibition of substrate phosphorylation by activated p38α.     

Discovery of a novel class of triazolones as Checkpoint Kinase inhibitors—Hit to lead exploration

Checkpoint Kinase-1 (Chk1, CHK1, CHEK1) is a Ser/Thr protein kinase that mediates cellular responses to DNA-damage. A novel class of Chk1 inhibitors, triazoloquinolones/triazolones (TZ’s) was identified by high throughput screening. The optimization of these hits to provide a lead series is described.