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Normal hepatocytes do not express endogenous uncoupling protein 2 (UCP2) in adult liver, although Kupffer cells do, and it is strikingly induced in hepatocytes in steatotic liver and obese conditions. However, the direct link of UCP2 with the pathogenic development of liver diseases and liver injury remains elusive. Here we report that targeted expression of UCP2 to mouse liver increases susceptibility to acute liver injury induced by lipopolysaccharide (LPS) and galactosamine (GalN). UCP2 appears to enhance proton leak, leading to mild uncoupling in a guanosine diphosphate-repressible manner. Indeed, mitochondria from the genetically manipulated mouse liver have increased state 4 respiration, lower respiratory control ratio, and reduced adenosine triphosphate (ATP) levels, which altered mitochondrial physiology. To address the underlying mechanism of how UCP2 and the reduced energy coupling efficiency enhance cell death in mouse liver, we show that the reduced ATP levels lead to activation of 5AMP-activated protein kinase (AMPK) and its downstream effector, c-Jun N-terminal kinase; thus, the increased sensitivity toward LPS/GalN-induces apoptosis. Importantly, we show that inhibition of UCP2 activity by its pharmacological inhibitor genipin prevents LPS/GalN-induced ATP reduction, AMPK activation, and apoptosis. Also, inhibition of ATP production by oligomycin promotes LPS/GalN-induced cell death both in vivo and in vitro. Conclusion: Our results clearly show that targeted expression of UCP2 in liver may result in compromised mitochondrial physiology that contributes to enhanced cell death and suggests a potential role of UCP2 in the development of liver diseases. (HEPATOLOGY 2009;50:1204-1216

Despite remarkable progress in the development of both viral and non-viral gene delivery vectors for cystic fibrosis therapy, low efficiency of gene transfer to the airway epithelium is a major obstacle to clinical application. Here we develop formulations that enhance cellular absorption of adenoviral vectors. We selected excipients from a panel of pharmaceutically acceptable com-pounds known to enhance drug absorption. Transduction efficiency of the virus in the presence of each ingredient was assessed in vitro and in vivo. Mannitol and chitosan substantially enhanced transduction efficiency in vitro and augmented expression in vivo by 4 and 8 log units, respectively. The most successful formulation (a blend of sucrose, mannitol, and Pluronic F68) transduced 100% of an A549 cell population in vitro and produced areas of intense gene expression in both large and small airways in vivo with minimal toxicity. Dose response studies also indicate that when placed in this formulation, the viral dose can be lowered by 1/2 log while maintaining superior levels of transgene expression. This formulation also enhanced the physical stability of the virus. No significant loss in titer was detected from a lyophilized formulation after storage at 25 degrees C for 30 days.

The occurrence of high resistance to carbofuran in the Chiayi and Changhua areas suggests that this compound should be replaced with chemicals having a different mode of action, such as chlorpyrifos, fipronil and permethrin, to which low cross-resistance has been detected.

No abstract

Aim The invasion pathways of pest arthropods can be traced using genetic tools to develop an understanding of the processes that have shaped successful invasions and to inform both pest management and conservation strategies in their non-native and native ranges, respectively. The redlegged earth mite, Halotydeus destructor, is a major economic pest in Australia, successfully establishing and spreading after arrival from South Africa more than 100 years ago. Halotydeus destructor has recently expanded its range and evolved resistance to numerous pesticides in Australia, raising questions around its origin and spread.Location South Africa and Australia.Methods We sampled H. destructor populations in South Africa and Australia and developed a microsatellite marker library. We then examined genetic variation using mtDNA and microsatellite markers across both native and invasive ranges to determine endemic genetic diversity within South Africa, identify the likely origin of invasive populations and test genetic divergence across Australia.Results The data show that H. destructor comprises a cryptic species complex in South Africa, with putative climatic/host plant associations that may correspond to regional variation. A lineage similar to that found near Cape Town has spread throughout Western and eastern Australia, where populations remain genetically similar.Main conclusions Tracing the invasion pathway of this economically important pest revealed cryptic lineages in South Africa which points to the need for a taxonomic revision. The absence of significant genetic structure across the wide invasive range of H. destructor within Australia has implications for the development (and spread) of pesticide resistance and also points to recent local adaptation in physiological traits.

The first two authors contributed equally to this work.The ontology of imidazoline receptors was first proposed by Bousquet P in 1980s [1] and was classified as I 1 , I 2 , and I 3 (non-I 1 /I 2 ) subtypes. However, deficiency in high selective ligand restrains the illumination of I 1 imidazoline receptor functions. In 2000, Piletz JE screened human hippocampal expression library and found a strong candidate protein for I 1 imidazoline receptor, named "imidazoline receptor antisera-selected" (IRAS) [2]. Concurrently, Alahari SK discovered a protein with identical sequence as IRAS and named it as "nischarin" [3]. Afterward, this protein was proved to be similar with I 1 imidazoline receptor in tissue distribution, ligand-binding property, and intracellular signallings and mediate many processes such as hypotensive effect of rilmenidine, inhibition of opioid addiction, inhibition of cell migration, antiapoptosis, and so on [4]. Herein, we originally generated IRAS conditional knockout mice (IRAS floxed/floxed ) and IRAS null mice (IRAS À/À ), which might be valuable tools for functional exploration of IRAS/nischarin and I 1 imidazoline receptors.Mouse IRAS gene is located on chromosome 14 and scatters into 21 exons, and we generated IRAS floxed/floxed mice by flanking exon 4 with loxP sites and then obtained IRAS À/À mice by crossing EIIa-Cre mice ( Figure 1A and Data S1 for more details). As for IRAS À/À mice, genotyping with the primer pair 5loxP-f and 3loxP-r could distinguish wild-type, heterozygote, and knockout mice ( Figure 1B). Sequencing result of the PCR product from 5loxP-f and 3flank primer pair revealed that only a loxP site remained between upstream and downstream homologous arms. In RTqPCR, the amplification curve of IRAS À/À was normal when the primer pair was originated from exon 3 and exon 5; however, the product is smaller than the wild type ( Figure 1C). Moreover, the amplification curve of IRAS À/À was null when the primer pair was originated from exon 2 and exon 4 ( Figure 1D). All the results above illustrated the normal transcription and the absence of exon 4 counterpart in the transcription product of IRAS À/À mice, which resulted in a new stop code in the following exon 5. In the Western blot assay, three bands, between 95 and 130 kD, were absent in cerebellum tissue of IRAS À/À mice ( Figure 1E). As several bands that represent functional IRAS have been reported, including 85 and 33 kD in human and 67 kD in bovine, these characteristic bands were proposed to be the splicing products in mouse cerebellum. All the above declared the functional deletion of IRAS gene.The IRAS À/À mice were born small compared with wild-type littermates (Figure 2A, B). Furthermore, weights of knockout embryos at day 12.5 (0.121 g, 0.133 g, and 0.143 g) were smaller than those of wild-type littermates (0.173 and 0.173 g) ( Figure 2C). These results suggested the participation of IRAS in 978 CNS Neuroscience & Therapeutics 19 (2013) 978-981 ª 2013 John Wiley & Sons Ltd prenatal growth and the postnatal growth re...

PEGylated adenoviruses are suitable gene delivery vehicles for oral administration.