Squalene Quantification Using Octadecylbenzene as the Internal Standard

AbstractSqualene was determined by HPLC using octadecylbenzene (ODB) as an internal standard. Squalene and ODB were monitored at 210nm. The retention times of squalene and ODB were 7.59 and 8.54min, respectively. Squalene was determined from the peak area ratios of squalene/ODB detected at 210nm. After treatment with 0.5M KOH containing ethanol at 90°C for 1h, squalene in the saponified lipid fraction was extracted using n-hexane. No interfering peak was observed. Linearity of this method was observed in the range 80–900 ng. ODB is useful as an internal standard for squalene determinations

On the Bio-Rearrangement into Fully Saturated Fatty Acids-Containing Triglyceride in Aurantiochytrium sp

AbstractA strain of Aurantiochytrium sp. was grown in media with various concentrations of glucose to monitor triglyceride production as a potential source of oil for biodiesel. The fatty acid composition of triglyceride in the strain was unique, because the fatty acids consisted of only 6 molecular species, and the major species were myristic, pentadecanoic, palmitic, heptadecanoic, docosapentaenoic, and docosahexaenoic acids. When cells were cultured in glucose-rich (over 9%) medium for 4 days, the triglyceride yields were 0.5-1.0g/L. After culture for 4 days, the fatty acid composition of triglyceride was nearly identical in all cells grown in media containing various concentrations of glucose. However, when cells were grown in medium containing 12% glucose for 12 days, unique triglyceride containing only saturated fatty acids accumulated. This bio-rearrangement into fully-saturated fatty acids-containing triglyceride may be utilized for the preparation of biodiesel oil

Succession of Genetic Diversity of Botryococcus braunii (Trebouxiophyceae) in Two Japanese Reservoirs

AbstractBotryococcus braunii is a green colonial alga that produces large volumes of liquid hydrocarbon. Therefore, B. braunii is expected to be useful as an alternative fuel resource. Natural blooms of B. braunii have been recorded in several lakes and reservoirs. Elucidation of natural B. braunii blooming would provide important information for the development of an open-pond cultivation system. In this study, we periodically assessed the genetic diversity and colony density of B. braunii populations, along with several environmental parameters, in two Japanese reservoirs (provisionally called “N” and “S”) from December 2008 to December 2009. Reservoir N had low numbers of B. braunii colonies whereas Reservoir S was characterized by periodic density increases that occurred in December 2008, and in March, September, and December 2009. Population genetics analysis using specific environmental sequences (PGA-SES method) was conducted for B. braunii populations for the first time. Among the B. braunii-dominated samples of Reservoir S, high levels of genetic diversity were observed in December 2008 and March 2009, whereas the diversity levels in September and December 2009 were low. The results suggest that B. braunii periodicity can be categorized into a high genetic diversity type and a low genetic diversity type. The high genetic diversity type may be caused by simultaneous growth of many genotypes, whereas the low genetic diversity type seems to be explained by increases in the cell density of only a few adapted genotypes

Diplophrys mutabilis sp. nov., a New Member of Labyrinthulomycetes from Freshwater Habitats

Diplophrys is a ubiquitous protist genus belonging to the class Labyrinthulomycetes. Although most members of Labyrinthulomycetes prefer marine habitats, the genus Diplophrys exclusively consists of the freshwater species Diplophrys archeri and Diplophrys parva. To investigate the genus Diplophrys, several novel strains were isolated from Japanese freshwater environments, and cultures of the strains were established. Among the strains, an organism isolated from Lake Nojiri displayed some characteristic features different from that of both D. archeri and D. parva. Thus, we described this strain as a new species, Diplophrys mutabilis. D. mutabilis can be cultured using dried water flea as food. This species had an orbicular to fusiform shape, and it occasionally penetrated prey with prominent cytoplasm. From a molecular phylogenetic analysis based on 18S rRNA sequences, D. mutabilis evidently belongs to Amphitremida, Labyrinthulomycetes. This study suggests that these species form a unique group in Labyrinthulomycetes

Desiccation tolerance of Botryococcus braunii (Trebouxiophyceae, Chlorophyta) and extreme temperature tolerance of dehydrated cells

Botryococcus braunii Kützing, a green colonial microalga, occurs worldwide in both freshwater and brackish water environments. Despite considerable attention to B. braunii as a potential source of renewable fuel, many ecophysiological properties of this alga remain unknown. Here, we examined the desiccation and temperature tolerances of B. braunii using two newly isolated strains BOD-NG17 and BOD-GJ2. Both strains survived through 6- and 8-month desiccation treatments but not through a 12-month treatment. Interestingly, the desiccation-treated cells of B. braunii gained tolerance to extreme temperature shifts, i.e., high temperature (40 °C) and freezing (−20 °C). Both strains survived for at least 4 and 10 days at 40 and −20 °C, respectively, while the untreated cells barely survived at these temperatures. These traits would enable long-distance dispersal of B. braunii cells and may account for the worldwide distribution of this algal species. Extracellular substances such as polysaccharides and hydrocarbons seem to confer the desiccation tolerance

Discovery of X-ray emission rom the distant lensing cluster of galaxies CL2236-04 at z = 0.552

X-ray emission from the distant lensing cluster CL2236-04 at $z$ = 0.552 was discovered by ASCA and ROSAT/HRI observations. If the spherical symmetric mass distribution model of the cluster is assumed, the lensing estimate of the cluster mass is a factor of two higher than that obtained from X-ray observations as reported for many distant clusters. However, the elliptical and clumpy lens model proposed by Kneib et al.(1993) is surprisingly consistent with the X-ray observations assuming that the X-ray emitting hot gas is isothermal and in a hydrostatic equilibrium state. The existence of the cooling flow in the central region of the cluster is indicated by the short central cooling time and the excess flux detected by ROSAT/HRI compared to the ASCA flux. However, it is shown that even if the AXJ2239-0429 has a cooling flow in the central region, the temperature measured by ASCA which is the mean emission-weighted cluster temperature in this case, should not be cooler than and different from the virial temperature of the cluster. Therefore, we conclude that the effect of the clumpiness and non-zero ellipticity in the mass distribution of the cluster are essential to explain the observed feature of the giant luminous arc, and there is no discrepancy between strong lensing and X-ray estimation of the mass of the cluster in this cluster.Comment: 18 pages, including 4 postscripts figs, LaTex. To appear in Part 1 of The Astrophysical Journa

Identification of intracellular squalene in living algae, Aurantiochytrium mangrovei with hyper-spectral coherent anti-Stokes Raman microscopy using a sub-nanosecond supercontinuum laser source

We applied hyper-spectral coherent anti-Stokes Raman scattering imaging to intracellular lipid identification in living microalgae, Aurantiochytrium mangrovei 18W-13a. Two different lipids, squalene and triacylglycerol, were found inside living cells with clear vibrational contrast. Based on the endogenous lipid band as a result of the cis C[DOUBLE BOND]C stretch vibrational mode, squalene and triacylglycerol were clearly distinguished in different intracellular areas. In particular, squalene was detected solely in vacuoles as lipid particles, which was also supported by electron microscopy

Pavlov's Cockroach: Classical Conditioning of Salivation in an Insect

Secretion of saliva to aid swallowing and digestion is an important physiological function found in many vertebrates and invertebrates. Pavlov reported classical conditioning of salivation in dogs a century ago. Conditioning of salivation, however, has been so far reported only in dogs and humans, and its underlying neural mechanisms remain elusive because of the complexity of the mammalian brain. We previously reported that, in cockroaches Periplaneta americana, salivary neurons that control salivation exhibited increased responses to an odor after conditioning trials in which the odor was paired with sucrose solution. However, no direct evidence of conditioning of salivation was obtained. In this study, we investigated the effects of conditioning trials on the level of salivation. Untrained cockroaches exhibited salivary responses to sucrose solution applied to the mouth but not to peppermint or vanilla odor applied to an antenna. After differential conditioning trials in which an odor was paired with sucrose solution and another odor was presented without pairing with sucrose solution, sucrose-associated odor induced an increase in the level of salivation, but the odor presented alone did not. The conditioning effect lasted for one day after conditioning trials. This study demonstrates, for the first time, classical conditioning of salivation in species other than dogs and humans, thereby providing the first evidence of sophisticated neural control of autonomic function in insects. The results provide a useful model system for studying cellular basis of conditioning of salivation in the simpler nervous system of insects