Recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis> produces tailor-made biopolyester granules for applications in fluorescence activated cell sorting: functional display of the mouse interleukin-2 and myelin oligodendrocyte glycoprotein

Background  

Fluorescence activated cell sorting (FACS) is a powerful technique for the qualitative and quantitative detection of biomolecules used widely in both basic research and clinical diagnostic applications. Beads displaying a specific antigen are used to bind antibodies which are then fluorescently labelled using secondary antibodies. As the individual suspension bead passes through the sensing region of the FACS machine, fluorescent signals are acquired and analysed. Currently, antigens are tediously purified and chemically cross-linked to preformed beads. Purification and coupling of proteins often renders them inactive and they will not be displayed in its native configuration. As an alternative, we genetically engineered Escherichia coli to produce biopolyester (polyhdroxyalkanoate=PHA) granules displaying diagnostically relevant antigens in their native conformation and suitable for FACS analysis.     

MicroRNAs show a wide diversity of expression profiles in the developing and mature central nervous system

Background  

MicroRNA (miRNA) encoding genes are abundant in vertebrate genomes but very few have been studied in any detail. Bioinformatic tools allow prediction of miRNA targets and this information coupled with knowledge of miRNA expression profiles facilitates formulation of hypotheses of miRNA function. Although the central nervous system (CNS) is a prominent site of miRNA expression, virtually nothing is known about the spatial and temporal expression profiles of miRNAs in the brain. To provide an overview of the breadth of miRNA expression in the CNS, we performed a comprehensive analysis of the neuroanatomical expression profiles of 38 abundant conserved miRNAs in developing and adult zebrafish brain.     

Seasonal monogamy and multiple paternity in a wild population of a territorial reptile (tuatara)

Investigating the mating system of a population provides insight into the evolution of reproductive patterns, and can inform conservation management of threatened or endangered species. Combining behavioural and genetic data is necessary to fully understand the mating system and factors affecting male reproductive success, yet behavioural data are often difficult to collect for threatened species. In the present study, we use behavioural data and paternity analyses to characterize the mating system of a high density population of a long-lived, ancient reptile (tuatara, Sphenodon punctatus ). We further investigate the phenotypic traits (including body size, body condition, tail length, and ectoparasite load) that affect male reproductive success. Our behavioural data reflect a seasonally monogamous system with low levels of polyandry and polygyny that are consistent with male mate guarding. Male reproduction is highly skewed (only 25–30% of males are successful), and body size is the primary predictor of male reproductive success. Based on the genetic data, multiple paternity was found in only 8% of clutches, and the results of the paternity analyses showed monandrous clutches from socially polyandrous females. Our behavioural and genetic results revealed complexities in female mating patterns that support the potential for cryptic female choice or sperm competition. This warrants further experimental investigation into the mechanisms underlying reptile fertilization and the disparities between social and genetic polyandry in wild populations.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 161–170.     

Availability of and access to critical habitats in regulated rivers: effects of low-head barriers on threatened lampreys

1.  Conservation of freshwater animal populations requires their access to, as well as sufficient availability of, critical habitats, such as those for reproduction. Abundant small-scale barriers may cause extensive fragmentation of freshwater habitat but, by comparison to larger structures their effects are rarely considered by catchment managers. The relationship between the distribution of, and access to, spawning habitat in a regulated river, characterized by abundant small barriers, was examined for river lamprey Lampetra fluviatilis , a threatened migratory fish.
2.  Telemetry of adult lamprey in the River Derwent, North East England was used to quantify upriver migration and access to spawning habitat, together with surveys of spawning habitat availability and spawning activity between 2002 and 2007.
3.  Access in to the Derwent appeared severely restricted by a tidal barrage, beyond which lamprey migrated rapidly in unobstructed reaches. Of all lamprey tagged in the lower 4 km of river, or ascending the barrage, 64% and 17% passed the first and second weirs respectively, with high flows crucial for this. Although over 98% of lamprey spawning habitat occurred more than 51 km upstream, on average just 1.8% of river lamprey spawners were recorded there.
4.  In order to protect or rehabilitate species or species assemblages, greater attention needs to be paid to the relative spatial distribution of low-head barriers and the resultant availability of key habitats within individual catchments. This is particularly important given the renewed emphasis internationally on low-head hydropower solutions as a source of renewable energy, and the rapid growth in numbers of low-head barriers in many catchments.     

Chemopreventive and renal protective effects for docosahexaenoic acid (DHA): implications of CRP and lipid peroxides

Background

The fish oil-derived ω-3 fatty acids, like docosahexanoic (DHA), claim a plethora of health benefits. We currently evaluated the antitumor effects of DHA, alone or in combination with cisplatin (CP) in the EAC solid tumor mice model, and monitored concomitant changes in serum levels of C-reactive protein (CRP), lipid peroxidation (measured as malondialdehyde; MDA) and leukocytic count (LC). Further, we verified the capacity of DHA to ameliorate the lethal, CP-induced nephrotoxicity in rats and the molecular mechanisms involved therein.

Results

EAC-bearing mice exhibited markedly elevated LC (2-fold), CRP (11-fold) and MDA levels (2.7-fold). DHA (125, 250 mg/kg) elicited significant, dose-dependent reductions in tumor size (38%, 79%; respectively), as well as in LC, CRP and MDA levels. These effects for CP were appreciably lower than those of DHA (250 mg/kg). Interestingly, DHA (125 mg/kg) markedly enhanced the chemopreventive effects of CP and boosted its ability to reduce serum CRP and MDA levels. Correlation studies revealed a high degree of positive association between tumor growth and each of CRP (r = 0.85) and leukocytosis (r = 0.89), thus attesting to a diagnostic/prognostic role for CRP. On the other hand, a single CP dose (10 mg/kg) induced nephrotoxicity in rats that was evidenced by proteinuria, deterioration of glomerular filtration rate (GFR, -4-fold), a rise in serum creatinine/urea levels (2–5-fold) after 4 days, and globally-induced animal fatalities after 7 days. Kidney-homogenates from CP-treated rats displayed significantly elevated MDA- and TNF-α-, but reduced GSH-, levels. Rats treated with DHA (250 mg/kg, but not 125 mg/kg) survived the lethal effects of CP, and showed a significant recovery of GFR; while their homogenates had markedly-reduced MDA- and TNF-α-, but -increased GSH-levels. Significant association was detected between creatinine level and those of MDA (r = 0.81), TNF-α ) r = 0.92) and GSH (r = -0.82); implying causal relationships.

Conclusion

DHA elicited prominent chemopreventive effects on its own, and appreciably augmented those of CP as well. The extent of tumor progression in various mouse groups was highly reflected by CRP levels (thus implying a diagnostic/prognostic role for CRP). Further, this study is the first to reveal that DHA can obliterate the lethal CP-induced nephrotoxicity and renal tissue injury. At the molecular level, DHA appears to act by reducing leukocytosis, systemic inflammation, and oxidative stress.     

Diet and social conditions during sexual maturation have unpredictable influences on female life history trade‐offs

The trade‐off between gametes and soma is central to life history evolution. Oosorption has been proposed as a mechanism by which females can redirect nutrients invested in oocytes into survival when conditions for reproduction are poor. Although positive correlations between oocyte degradation and lifespan have been documented in oviparous insects, the adaptive significance of this process in species with more complex reproductive biology has not been explored. Further, environmental condition is a multivariate state, and combinations of environmental stresses may interact in unpredictable ways. Previous work on the ovoviviparous cockroach, Nauphoeta cinerea, revealed that females manipulated to mate late relative to sexual maturation experience age‐related loss in fecundity because of loss of viable oocytes via apoptosis. This loss in fecundity is correlated with a reduction in female mate choice. Food deprivation while mating is delayed further increases levels of oocyte apoptosis, but the relationship between starvation‐induced apoptosis and life history are unknown. To investigate this, virgin females were either fed or starved from eclosion until provided with a mate at a time known to be suboptimal for fertility. Following mating, females were fed for the duration of their lifespan. We measured lifetime reproductive performance. Contrary to predictions, under conditions of delayed mating opportunity, starved females had greater fecundity, gave birth to more high‐quality offspring and had increased longevity compared with that of fed females. We suggest that understanding proximal mechanisms underlying life history trade‐offs, including the function of oocyte apoptosis, and how these mechanisms respond to varied environmental conditions is critical.     

Phylogenetics and biogeography of the parasitic genus Thesium L. (Santalaceae), with an emphasis on the Cape of South Africa

Thesium is a large genus of parasitic shrubs belonging to tribe Thesieae of Santalaceae. It has a principally Old World distribution, with the greatest diversity being found in southern Africa. Little is known about the relationships within Thesium or its relationships with its closest relatives. In this article, we present a first estimate of species‐level phylogenetic relationships in Thesium based on internal transcribed spacer (ITS) and trnL–trnF sequence data, and use this to explore the biogeographical history of the group. One hundred and four samples representing 72 Thesium spp. were included in a phylogenetic analysis. Plastid and combined data resolve Thesium as paraphyletic relative to Thesidium and Austroamericium with high posterior probability and bootstrap support. ITS sequence data place Thesidium as sister to a large Thesium clade, but with weak support. Ancestral range reconstruction and dating analysis suggest a southern African origin for the group, with a crown age of 39.1 ± 11.9 Mya, followed by dispersal into Europe and South America. A large clade of Cape species split in the Miocene from a clade comprising tropical species (25.5 ± 7.3 Mya) with the diversification of extant species beginning at 16.7 ± 6.3 Mya. © 2010 The Linnean Society of London, Botanical Journal of the Linnean Society, 2010, 162 , 435–452.     

毛乌素沙地南缘沙丘生物结皮中微生物分布特征

为探明半干旱沙区生物结皮中微生物分布特征,对毛乌素沙地南缘沙丘生物结皮中微生物数量进行了测定。结果表明:微生物总数从丘顶到丘间地呈递增趋势,除丘顶与迎风坡、迎风坡与背风坡结皮层微生物总数差异不显著外,其他各地貌部位结皮层微生物数量之间差异显著。同一地貌部位结皮层、0～5和5～10cm土层微生物垂直分布有变化,其变化规律为:除迎风坡放线菌数量呈先增加后递减、迎风坡微生物总数、细菌、真菌和丘顶真菌数量随剖面的加深呈递减外,其他各地貌部位微生物数量均呈先降低,后增加的趋势。微生物类群的组成表现为细菌最多,放线菌次之,真菌最少。在丘间地细菌所占微生物总数的比例与丘顶相比有所增加,而放线菌和真菌的比例有所减少。结皮下0～5和5～10cm土层微生物分布与土壤含水量的变化同步,说明土壤水分可能是影响微生物垂直分布的重要因子。     

Generation of medaka gene knockout models by target-selected mutagenesis

We have established a reverse genetics approach for the routine generation of medaka (Oryzias latipes) gene knockouts. A cryopreserved library of N-ethyl-N-nitrosourea (ENU) mutagenized fish was screened by high-throughput resequencing for induced point mutations. Nonsense and splice site mutations were retrieved for the Blm, Sirt1, Parkin and p53 genes and functional characterization of p53 mutants indicated a complete knockout of p53 function. The current cryopreserved resource is expected to contain knockouts for most medaka genes.