Fatty acid synthase modulates homeostatic responses to myocardial stress

Fatty acid synthase (FAS) promotes energy storage through de novo lipogenesis and participates in signaling by the nuclear receptor PPARα in noncardiac tissues. To determine if de novo lipogenesis is relevant to cardiac physiology, we generated and characterized FAS knockout in the myocardium (FASKard) mice. FASKard mice develop normally, manifest normal resting heart function, and have normal cardiac PPARα signaling as well as fatty acid oxidation. However, they decompensate with stress. Most die within 1 h of transverse aortic constriction, probably due to arrhythmia. Voltage clamp measurements of FASKard cardiomyocytes show hyperactivation of L-type calcium channel current that could not be reversed with palmitate supplementation. Of the classic regulators of this current, Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) but not protein kinase A signaling is activated in FASKard hearts, and knockdown of FAS in cultured cells activates CaMKII. In addition to being intolerant of the stress of acute pressure, FASKard hearts were also intolerant of the stress of aging, reflected as persistent CaMKII hyperactivation, progression to dilatation, and premature death by ～1 year of age. CaMKII signaling appears to be pathogenic in FASKard hearts because inhibition of its signaling in vivo rescues mice from early mortality after transverse aortic constriction. FAS was also increased in two mechanistically distinct mouse models of heart failure and in the hearts of humans with end stage cardiomyopathy. These data implicate a novel relationship between FAS and calcium signaling in the heart and suggest that FAS induction in stressed myocardium represents a compensatory response to protect cardiomyocytes from pathological calcium flux.     

The Drosophila gene disruption project: progress using transposons with distinctive site specificities

The Drosophila Gene Disruption Project (GDP) has created a public collection of mutant strains containing single transposon insertions associated with different genes. These strains often disrupt gene function directly, allow production of new alleles, and have many other applications for analyzing gene function. Here we describe the addition of ～7600 new strains, which were selected from >140,000 additional P or piggyBac element integrations and 12,500 newly generated insertions of the Minos transposon. These additions nearly double the size of the collection and increase the number of tagged genes to at least 9440, approximately two-thirds of all annotated protein-coding genes. We also compare the site specificity of the three major transposons used in the project. All three elements insert only rarely within many Polycomb-regulated regions, a property that may contribute to the origin of "transposon-free regions" (TFRs) in metazoan genomes. Within other genomic regions, Minos transposes essentially at random, whereas P or piggyBac elements display distinctive hotspots and coldspots. P elements, as previously shown, have a strong preference for promoters. In contrast, piggyBac site selectivity suggests that it has evolved to reduce deleterious and increase adaptive changes in host gene expression. The propensity of Minos to integrate broadly makes possible a hybrid finishing strategy for the project that will bring >95% of Drosophila genes under experimental control within their native genomic contexts.     

Experimental use of two standard tick collection methods to evaluate the relative effectiveness of several plant-derived and synthetic repellents against Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae)

We used two standard tick collection methods to test the relative effectiveness of two natural product compounds (nootkatone and carvacrol, classified as an eremophilene sesquiterpene and a monoterpene, respectively, that are derived from botanical sources) with commercially-available plant-derived (EcoSMART Organic Insect Repellent, comprised of plant essential oils) and permethrin-based (Repel Permanone) repellents against Ixodes scapularis Say and Amblyomma americanum (L.). Cloth drags were equally effective in sampling both species of host-seeking nymphs, whereas CO, traps attracted primarily A. americanum. All four repellents performed well on drags, with nootkatone and Permanone Repel (100% repelled through 14 d) slightly more effective than carvacrol and EcoSMART (90.7% and 97.7% repelled at 14 d, respectively) at repelling I. scapularis nymphs. Although the same trend in percent repellency was noted in the CO2 trap trial against both A. americanum nymphs and adults, EcoSMART outperformed Permanone in repelling A. Americanum nymphs after 14 d in the drag trial. Generally, the effectiveness of all repellents tested declined over time. The use of tick drags and CO2 traps was rapid, inexpensive, and easy to use in determining the relative effectiveness of repellents in the field.     

Suppression of host-seeking Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae) nymphs after dual applications of plant-derived acaricides in New Jersey

We evaluated the ability of dual applications of natural, plant-derived acaricides to suppress nymphal Ixodes scapularis Say and Amblyomma americanum (L.) (Acari: Ixodidae) in a Lyme disease endemic area of New Jersey. An aqueous formulation of 2% nootkatone provided >90% control of I. scapularis through 7 d. Control declined to 80.9% at 14 d, and a second application was made that provided >95% control through the remaining 4 wk of the nymphal season. Nootkatone provided >90% control of A. americanum through 35 d postapplication. Applications of 2% carvacrol and EcoTrol T&O resulted in rapid knockdown of both tick species, but control declined significantly to 76.7 and 73.7%, respectively, after 14 d when a second application was made that extended control of both tick species to between 86.2 and 94.8% at 21 d. Subsequently, control declined steadily in all plots by 42 d postapplication except for I. scapularis in carvacrol-treated plots, where levels of control >90% were observed through 35 d. Of the three compounds tested, 2% nootkatone provided the most consistent results, with 96.5 and 91.9% control of I. scapularis and A. americanum through 42 and 35 d, respectively. The ability of plant-derived natural products to quickly suppress and maintain significant control of populations of these medically important ticks may represent a future alternative to the use of conventional synthetic acaricides. In addition, the demonstrated efficacy of properly-timed backpack sprayer application may enable homeowner access to these minimal-risk acaricides.     

Complex V TMEM70 deficiency results in mitochondrial nucleoid disorganization

Mutations in the TMEM70 gene are responsible for a familial form of complex V deficiency presenting with 3-methylglutaconic aciduria, lactic acidosis, cardiomyopathy and mitochondrial myopathy. Here we present a case of TMEM70 deficiency due to compound heterozygous mutations, who displayed abnormal mitochondria with whorled cristae in muscle. Immunogold electron microscopy and tomography shows for the first time that nucleoid clusters of mtDNA are disrupted in the abnormal mitochondria, with both nucleoids and mitochondrial respiratory chain complexes confined to the outer rings of the whorls. This could explain the differential effects on the expression and assembly of complex V in different tissues.     

Large‐scale bioenergy from additional harvest of forest biomass is neither sustainable nor greenhouse gas neutral

Owing to the peculiarities of forest net primary production humans would appropriate ca. 60% of the global increment of woody biomass if forest biomass were to produce 20% of current global primary energy supply. We argue that such an increase in biomass harvest would result in younger forests, lower biomass pools, depleted soil nutrient stocks and a loss of other ecosystem functions. The proposed strategy is likely to miss its main objective, i.e. to reduce greenhouse gas (GHG) emissions, because it would result in a reduction of biomass pools that may take decades to centuries to be paid back by fossil fuel substitution, if paid back at all. Eventually, depleted soil fertility will make the production unsustainable and require fertilization, which in turn increases GHG emissions due to N₂O emissions. Hence, large‐scale production of bioenergy from forest biomass is neither sustainable nor GHG neutral.     

Types, levels and patterns of low-copy DNA sequence divergence, and phylogenetic implications, for Gossypium genome types

To explore types, levels and patterns of genetic divergence among diploid Gossypium (cotton) genomes, 780 cDNA, genomic DNA and simple sequence repeat (SSR) loci were re-sequenced in Gossypium herbaceum (A1 genome), G. arboreum (A2), G. raimondii (D5), G. trilobum (D8), G. sturtianum (C1) and an outgroup, Gossypioides kirkii. Divergence among these genomes ranged from 7.32 polymorphic base pairs per 100 between G. kirkii and G. herbaceum (A1) to only 1.44 between G. herbaceum (A1) and G. arboreum (A2). SSR loci are least conserved with 12.71 polymorphic base pairs and 3.77 polymorphic sites per 100 base pairs, whereas expressed sequence tags are most conserved with 3.96 polymorphic base pairs and 2.06 sites. SSR loci also exhibit the highest percentage of 'extended polymorphisms' (spanning multiple consecutive nucleotides). The A genome lineage was particularly rapidly evolving, with the D genome also showing accelerated evolution relative to the C genome. Unexpected asymmetry in mutation rates was found, with much more transition than transversion mutation in the D genome after its divergence from a common ancestor shared with the A genome. This large quantity of orthologous DNA sequence strongly supports a phylogeny in which A-C divergence is more recent than A-D divergence, a subject that is of much importance in view of A-D polyploid formation being key to the evolution of the most productive and finest-quality cottons. Loci that are monomorphic within A or D genome types, but polymorphic between genome types, may be of practical importance for identifying locus-specific DNA markers in tetraploid cottons including leading cultivars.     

Effect of subtalar arthroereisis on the tibiotalar contact characteristics in a cadaveric flatfoot model

Previous studies reported the effect of flatfoot deformity on tibiotalar joint contact characteristics. The lateral shift of the load which occurred in flatfeet may be responsible for degenerative changes in the ankle joint. The purpose was to assess the pattern of joint contact stress of the tibiotalar joint in intact, flat, and corrected specimens with subtalar arthroereisis. Seven fresh-frozen cadaver specimens were studied in the intact and flat-footed condition after transection of ligaments which support the medial arch. Ankle joint contact stress and plantar pressure patterns were determined from a capacitive pressure sensor inserted in the tibiotalar joint and a pressure distribution platform when the specimens were axially loaded in simulated mid-stance. Contact pressure was also assessed after subtalar arthroereisis with a 12 mm Kalix implant for correction of the flatfoot deformity. A maximum contact pressure of 1414.2±319.2 kPa was recorded in the middle-medial region in intact specimens. Flatfoot caused a lateral shift in the pressure distribution (p<0.05). In the flat specimens, the maximum contact pressure of 1394.7 8±470.5 kPa was in the anterior-central region. After subtalar arthroereisis with the Kalix implant for correction of the flatfoot deformity a maximum contact pressure of 1323.3±497.5 kPa was observed in the middle-lateral region. In a cadaver model, subtalar arthroereisis with Kalix implant failed to restore a normal intraarticular ankle joint pressure pattern. Further interventions should be considered to restore a normal pressure pattern.