Plant regeneration from patchouli protoplasts encapsulated in alginate beads

Mesophyll protoplasts were isolated from leaves of in vitro grown patchouli (Pogostemon cablin Benth.). The protoplasts were encapsulated in alginate beads, approximately 2–3×10³ protoplasts per 25 l bead. Successful colony formation was induced when the protoplast beads were inoculated into a liquid medium supplemented with 10^-6 M NAA and 10^-5 M BA. The frequency of colony formation was improved greatly by the inclusion of several beads per ml medium. To induce high colony formation for a single bead, it was essential to culture protoplasts in the presence of nurse beads containing actively-growing cells of the same species. Rapid regeneration of plants from protoplast-derived calluses was accomplished by a two-step culture procedure with liquid and then solid media. Gas-chromatographic analyses showed that regenerated plants produced an essential oil comprising a full-set of patchouli sesquiterpenes.Abbreviations BA 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - f. wt. fresh weight - GC gas chromatography - MES 2-(N-morpholino)ethanesulfonic acid - NAA 1-naphthaleneacetic acid     

The Short‐Term Effect of Diacylglycerol Oil Consumption on Total and Dietary Fat Utilization in Overweight Women

Diacylglycerol (DAG) is a natural component of edible oils with metabolic characteristics distinct from those of triacylglycerol (TAG). Consumption of DAG oil (containing >80% DAG) induces greater fat oxidation than consumption of TAG oil. We compared the effects of 4 days of DAG oil consumption with those of TAG oil consumption on total and dietary fat oxidation over 24 h in overweight women using a whole‐room respiratory chamber. Overweight (BMI (kg/m²) ≥25) females participated in this double‐blind, crossover‐controlled trial. The subjects consumed test diets containing either TAG or DAG oil as 15% of their total caloric intake (mean test oil intake was 33.0 ± 3.1 g/day) during each 4‐day treatment. Fat oxidation and energy expenditure were measured in a respiratory chamber on the 4th day of each treatment. Compared with TAG oil, DAG oil consumption significantly increased total fat oxidation and dietary fat oxidation in overweight subjects. Total energy expenditure (TEE) and carbohydrate (CHO) oxidation did not significantly differ between DAG oil and TAG oil consumption in overweight subjects. Compared with TAG oil, DAG oil consumption enhanced total fat oxidation and dietary fat oxidation in overweight subjects. The enhanced fat metabolism in overweight subjects that consumed DAG oil partly explains the greater loss of body weight and body fat related to DAG oil consumption in weight‐loss studies.     

Production of polysaccharides by liquid cultures of Polianthes tuberosa cells; high production by reduction in the viscosity of culture medium

Summary To improve the production of extracellular polysaccharides (EPS) in liquid cultures of Polianthes tuberosa (tuberose) cells, the viscosity of the culture medium was lowered by addition of mineral salts. In cultures in the medium supplemented with 30 mM CaCl2, higher production of EPS (6.5g/l) has been realized (vs. 4.6 g/l without CaCl2).     

A Novel Subtype of Astrocytes Expressing TRPV4 (Transient Receptor Potential Vanilloid 4) Regulates Neuronal Excitability via Release of Gliotransmitters

Astrocytes play active roles in the regulation of synaptic transmission. Neuronal excitation can evoke Ca²⁺ transients in astrocytes, and these Ca²⁺ transients can modulate neuronal excitability. Although only a subset of astrocytes appears to communicate with neurons, the types of astrocytes that can regulate neuronal excitability are poorly characterized. We found that ∼30% of astrocytes in the brain express transient receptor potential vanilloid 4 (TRPV4), indicating that astrocytic subtypes can be classified on the basis of their expression patterns. When TRPV4⁺ astrocytes are activated by ligands such as arachidonic acid, the activation propagates to neighboring astrocytes through gap junctions and by ATP release from the TRPV4⁺ astrocytes. After activation, both TRPV4⁺ and TRPV4⁻ astrocytes release glutamate, which acts as an excitatory gliotransmitter to increase synaptic transmission through type 1 metabotropic glutamate receptor (mGluR). Our results indicate that TRPV4⁺ astrocytes constitute a novel subtype of the population and are solely responsible for initiating excitatory gliotransmitter release to enhance synaptic transmission. We propose that TRPV4⁺ astrocytes form a core of excitatory glial assembly in the brain and function to efficiently increase neuronal excitation in response to endogenous TRPV4 ligands.     

Development of wrapped liposomes: Novel liposomes comprised of polyanion drug and cationic lipid complexes wrapped with neutral lipids

Novel wrapped liposomes comprised of polyanion drug and cationic lipid complexes wrapped with neutral lipids were prepared using an efficient, innovative procedure. In this study, dextran fluorescein anionic (DFA) was used as an example of a polyanionic compound. During the process, neutral lipids accumulated around the complexes and eventually covered the complexes. The resulting liposomes were 120-140 nm in diameter and the encapsulation efficiency was up to 90%. In fetal bovine serum, DFA/cationic lipid complexes degraded rapidly but the wrapped liposomes were considerably more stable. Following intravenous administration to rats, DFA/cationic lipid complexes were rapidly eliminated whereas the wrapped liposomes exhibited a much longer blood half-life. These data suggest that DFA is located on the surface of the complexes, but DFA is present inside the wrapped liposomes. The drug-delivery properties of the wrapped liposomes established in the present study suggests that formulations based on this technology could offer important advantages for the administration of many types of drug including antisense oligonucleotides, plasmids and siRNAs which may therefore lead to improved therapeutic effectiveness of this range of drugs. The method of preparation of the wrapped liposomes is so simple that it should be straightforward to adapt to a manufacturing scale.     

Influence of prey on developmental performance, reproduction and prey consumption of Neoseiulus californicus (Acari: Phytoseiidae)

The Spical strain of the predatory mite Neoseiulus californicus (McGregor) is used as a biological control agent, but little is known about its preferred prey and host plants in Japan. Here we studied the development, reproduction and prey consumption of the Spical strain when fed on eggs of five different spider mite species deposited on both their laboratory-rearing plant and cherry, on which all five spider mite species developed well. The developmental periods of immature N. californicus females and males were significantly affected by the prey species they fed on, but not by the plants. No difference was found between males and females. The developmental period was shorter on eggs of two Tetranychus species than on eggs of Panonychus ulmi. Immature females had a higher predation rate than immature males. Preoviposition period, oviposition period and the number of eggs laid per female were not significantly affected by either the plants or the type of prey eggs. The postoviposition period and total adult longevity were shorter on eggs of P. ulmi than of the other four prey species, but there was no effect of plant substrate. The postoviposition period of the Spical strain was much longer than that of other N. californicus strains or other predatory mite species: the postoviposition period of the Spical strain was more than three times longer than the oviposition period, accounting for more than 75% of the total adult longevity. This suggests that the females need multiple mating to reach full egg load, but this remains to be tested. Total consumption by N. californicus adults was lower for eggs of P. ulmi than for eggs of the other four species, apparently because of the shorter postoviposition period when fed on eggs of P. ulmi. The intrinsic rates of natural increase (r _m) on the rearing plant did not differ among prey species, whereas those on cherry were significantly different: the value was higher on Tetranychus urticae eggs than on eggs of other species. Only when N. californicus fed on T. urticae eggs, the r _m-values were significantly different between the rearing plant and cherry (higher on cherry). Thus, the Spical strain of N. californicus could feed on eggs of all five spider mite species, deposited on a variety of plants with similar r _m-values, suggesting that it could be successfully used to control spider mites in orchards and various crop fields of Japan.     

The Synthesis of Dilnethyl α-Sorigenin

1,6,8-Trimethoxy-3-hydroxymethyl-2-naphthoic acid lactone (IV) was synthesized from benzoic acid in 21 steps. This lactone (IV) was completely identical with authentic dimethyl α-sorigenin, obtained by the methylation of natural α-sorigenin. Herewith the structure of α-sorigenin was confirmed to be 1,8-dihydroxy-6-methoxy-3-hydroxymethyl-2-naphthoic acid lactone (III).     

Up-regulation of glyceraldehyde-3-phosphate dehydrogenase gene expression by HIF-1 activity depending on Sp1 in hypoxic breast cancer cells

Hypoxia up-regulates the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in a cell type-specific manner. It is unknown whether this occurs in breast cancer. Here, we report that hypoxia up-regulates the GAPDH gene expression through breast cancer-specific molecular mechanisms in MCF-7 cells. Mutation analysis identified a novel hypoxia response element (HRE), in addition to the HRE found previously in prostate cancer LNCaP cells. Knockdown and overexpression of hypoxia-inducible factor (HIF)-1α indicated that HIF-1 contributed to the up-regulation of GAPDH gene expression by hypoxia. Although chromatin immunoprecipitation (ChIP) and plasmid immunoprecipitation analyses revealed the presence of HIF-1α on the novel HRE in both hypoxic cell lines, a mutation in either the novel HRE or its 3′-flanking GC-box resulted in a reduction of hypoxia-increased GAPDH promoter activity only in MCF-7 cells. ChIP analysis showed that Sp1 bound to the GC-box in MCF-7 cells, but not in LNCaP cells, in normoxia and hypoxia. Knockdown of Sp1 reduced hypoxia-increased promoter activity and expression level of GAPDH in MCF-7 cells. These results indicate that in MCF-7 cells, the activation of HIF-1 on the novel HRE contributes to the breast cancer-specific hypoxic induction of GAPDH gene expression and absolutely depends on the presence of Sp1 on the GC-box.