Use of online-dual-column extraction in conjunction with chiral liquid chromatography tandem mass spectrometry for determination of terbutaline enantiomers in human plasma

An online sample extraction chiral bioanalytical method was developed and validated for the quantification of terbutaline, a beta2-selective adrenoceptor agonist, spiked into human plasma by using two extraction columns and a chiral stationary phase (CSP) in conjunction with liquid chromatography tandem mass spectrometry (LC-MS/MS). In this method, two Oasis HLB extraction columns were used in parallel for plasma sample purification and a Chirobiotic T CSP was used for enantiomeric separation. Atmospheric pressure chemical ionization MS/MS was employed in multiple reaction monitoring mode for the detection and quantification. Subsequent to the addition of an internal standard solution, the plasma samples were directly injected onto the system for extraction and analysis. This method allowed the use of one of the extraction columns for purification while the other was being equilibrated. Hence, the time required for reconditioning the extraction columns did not contribute to the total analysis time per sample, which resulted in a shorter run time and higher throughput. A lower limit of quantification of 1.0 ng/mL was achieved using only 50 microliter of human plasma. The method was validated with a dynamic range of 1.0-200 ng/mL. The intra- and interday precision was no more than 11% CV and the assay accuracy was between 94-106%.     

Experimental demonstration of close-range olfaction and contact chemoreception in the Brazilian harvestman, Iporangaia pustulosa

We studied the ability to detect food by close-range olfaction and contact chemoreception in the harvestman Iporangaia pustulosa Mello-Leitão (Opiliones: Laniatores: Gonyleptidae). We first tested the reaction of individuals towards tasteless (pure agar), aversive (agar with salt), and food-intake stimulating substrates (agar with saccharose). Only the substrate containing saccharose was consumed. Contact (mainly with legs II) was necessary for detection of the agar and, before ingestion, the stimulus was always tapped with legs I. In the second experiment, we observed the behavior of individuals in an arena with a screened plastic box containing pieces of Tenebrio molitor L. (Coleoptera: Tenebrionidae) larvae. Individuals spent more time on the box containing food than on the control. In the third experiment, in an arena identical to that used in Experiment 2, we introduced a live but motionless T. molitor larva in the box. There was no difference between experimental and control treatments. We also observed the behavior of I. pustulosa in an arena containing live isopods. In first capture attempts, isopods were only detected upon contact, mainly with legs I. Our results suggest that (i) I. pustulosa is capable of detecting food only by its chemical properties; (ii) food with weak odor may not be detected by close-range olfaction; and (iii) legs I and II are important for food detection but, before ingestion, legs I are used to examine potential food items.     

What do tadpoles really eat? Assessing the trophic status of an understudied and imperiled group of consumers in freshwater habitats

1. Understanding the trophic status of consumers in freshwater habitats is central to understanding their ecological roles and significance. Tadpoles are a diverse and abundant component of many freshwater habitats, yet we know relatively little about their feeding ecology and true trophic status compared with many other consumer groups. While many tadpole species are labelled herbivores or detritivores, there is surprisingly little evidence to support these trophic assignments. 2. Here we discuss shortcomings in our knowledge of the feeding ecology and trophic status of tadpoles and provide suggestions and examples of how we can more accurately quantify their trophic status and ecological significance. 3. Given the catastrophic amphibian declines that are ongoing in many regions of the planet, there is a sense of urgency regarding this information. Understanding the varied ecological roles of tadpoles will allow for more effective conservation of remaining populations, benefit captive breeding programmes, and allow for more accurate predictions of the ecological consequences of their losses.     

N-linked glycosylation of the superoxide-producing NADPH oxidase Nox1

Nox1 is a membrane-integrated protein that belongs to the Nox family of superoxide-producing NADPH oxidases. Here we show that human Nox1 undergoes glycosylation at Asn-162 and Asn-236 in the second and third extracellular loops, respectively. Simultaneous threonine substitution for these residues completely abrogates the glycosylation, but does not prevent Nox1 from forming a heterodimer with p22^phox, trafficking to the cell surface, or producing superoxide. In the absence of p22^phox, Nox1 is transported to the plasma membrane mainly as a form with high mannose N-glycans, although their conversion into complex N-glycans is induced by expression of p22^phox. These findings indicate that glycosylation and subsequent N-glycan maturation of Nox1 are both dispensable for its cell surface recruitment. Superoxide production by unglycosylated Nox1 is largely dependent on p22^phox, which is abrogated by glutamine substitution for Pro-156 in p22^phox, a mutation leading to a defective interaction with the Nox1-activating protein Noxo1. Thus p22^phox directly contributes to Nox1 activation in a glycosylation-independent manner, besides its significant role in Nox1 glycan maturation.     

丛枝菌根真菌侵染番茄离体毛状根双重培养体系的建立

利用发根农杆菌Agrobacterium rhizogenes A4诱导樱桃番茄Micro-Tom产生毛状根,并在此基础上成功建立了丛枝菌根(AM)真菌根内球囊霉Glomus intraradices与转移Ri T-DNA番茄离体毛状根的双重培养体系。该真菌侵染14d左右,菌丝开始形成多级分支,17-21d时,一些菌丝顶端膨大,长出新生孢子。接种后3个月时,每皿孢子数量达到600-800个。新形成的孢子无需休眠,可直接侵染番茄离体根。成功的番茄双重培养为番茄菌根生理分子机制的研究提供了理想的实验体系。     

Effects of omnivorous filter‐feeding fish and nutrient enrichment on the plankton community and water transparency of a tropical reservoir

1. The major aim of this study was to test the hypothesis that nutrient enrichment and the introduction of the Nile tilapia (Oreochromis niloticus), an exotic omnivorous filter‐feeding fish, operate interdependently to regulate plankton communities and water transparency of a tropical reservoir in the semi‐arid northeastern Brazil. 2. A field experiment was performed for 5 weeks in 20 enclosures (9.8 m³) to which four treatments were randomly allocated: tilapia addition (F), nutrient addition (N), tilapia and nutrient addition (F + N) and a control treatment with no tilapia or nutrient addition (C). A two‐way repeated measures anova was undertaken to test for time, tilapia and nutrient effects and their interactions on water transparency, total phosphorus and total nitrogen concentrations, phytoplankton biovolume and zooplankton biomass. 3. Nutrient addition had no effect except on rotifer biomass, but there were significant fish effects on the biomass of total zooplankton, copepod nauplii, rotifers, cladocerans and calanoid copepods and on the biovolume of total phytoplankton, large algae (GALD ≥ 50 μm), Bacillariophyta and Zygnemaphyceae and on Secchi depth. In addition, we found significant interaction effects between tilapia and nutrients on Secchi depth and rotifers. Overall, tilapia decreased the biomass of most zooplankton taxa and large algae (diatoms) and decreased water transparency, while nutrient enrichment increased the biomass of rotifers, but only in the absence of tilapia. 4. In conclusion, the influence of fish on the reservoir plankton community and water transparency was significant and even greater than that of nutrient loading. This suggests that biomanipulation of filter‐feeding tilapias may be of importance for water quality management of eutrophic reservoirs in tropical semi‐arid regions.     

A genetic analysis of the Sakishima islanders reveals no relationship with Taiwan aborigines but shared ancestry with Ainu and main‐island Japanese

The Sakishima islands are members of the Ryukyu island chain, stretching from the southwestern tip of the Japanese archipelago to Taiwan in the East China Sea. Archaeological data indicate cultural similarities between inhabitants of prehistoric Sakishima and Neolithic Taiwan. Recent studies based on tooth crown traits show remarkably high inter‐island diversity among Ryukyu islanders, suggesting that the Sakishima islanders might have genetic backgrounds distinct from main‐island Okinawa people. To investigate the genetic diversity of the Ryukyu islanders, we analyzed mtDNA, Y chromosome, and autosomal short tandem repeat loci in a sample of main‐island Okinawa people and Sakishima (Miyako and Ishigaki) islanders whose participated in a previous study of tooth crown morphology. Our phylogenetic analysis of maternal (mtDNA) and paternal (Y chromosome) lineages shows that the Sakishima islanders are more closely related to people from the Japanese archipelago than to Taiwan aborigines. Miyako islanders and the Hokkaido Ainu have the first and second highest frequencies (respectively) of the Y‐chromosomal Alu‐insertion polymorphism, which is a presumable Jomon marker. Genetic diversity statistics show no evidence of demographic reduction or of extreme isolation in each island's population. Thus, we conclude that 1) Neolithic expansion from Taiwan did not contribute to the gene pool of modern Sakishima islanders, 2) male‐lineage of the Ryukyu islanders likely shares a common ancestor with the Hokkaido Ainu who are presumably direct descendants of the Jomon people, and 3) frequent reciprocal gene flow among islands has masked the trace of common ancestry in the Ryukyu island chain. Am J Phys Anthropol, 2010. © 2010 Wiley‐Liss, Inc.     

Genetic basis and breeding application of clonal diversity and dual reproduction modes in polyploid Carassius auratus gibelio

A unisexual species is generally associated with polyploidy, and reproduced by a unisexual reproduction mode, such as gyno- genesis, hybridogenesis or parthenogenesis. Compared with other unisexual and polyploid species, gibel carp (Carassius au- ratus gibelio) has a higher ploidy level of hexaploid. It has undergone several successive rounds of genome polyploidy, and experienced an additional, more recent genome duplication event. More significantly, the dual reproduction modes, including gynogenesis and s...     

Influence of castration-induced testosterone and estradiol deficiency on obesity and glucose metabolism in male Göttingen minipigs

Low testosterone and estradiol concentrations are predictive for the development of the metabolic syndrome in men and women, respectively. The aim of this study was to investigate the influence of sex hormone deficiency on food intake, body weight, body composition and glucose metabolism in male Göttingen minipigs.Five adult male Göttingen minipigs were studied before castration (pre-cast), 10-18 days (post-cast 1) and 10-11 weeks (post-cast 2) after castration. Parameters of interest were food intake, body weight, body fat percentage and sex hormone concentrations. Furthermore glucose tolerance, glucagon suppression, insulin resistance, beta cell function and disposition index were evaluated by oral and intravenous glucose tolerance tests.Castration led to almost complete disappearance of circulating testosterone and estradiol and secondarily to increased food intake, body weight and body fat percentage. Ten-eighteen days sex hormone deficiency (post-cast 1) did not significantly change any of the investigated metabolic parameters compared to pre-cast levels. Ten weeks after castration (post-cast 2) significant insulin resistance, glucose intolerance and hyperglucagonemia was found, and the beta cell function and the disposition index both were decreased.In conclusion, castration-induced sex hormone deficiency in male Göttingen minipigs results in hyperphagia, obesity and disturbed glucose metabolism, which are some of the features typical for the human metabolic syndrome.