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Mitochondrial genomes (mitochondrial DNA, mtDNA) encode essential oxidative phosphorylation (OXPHOS) components. Because hundreds of mtDNAs exist per cell, a deletion in a single mtDNA has little impact. However, if the deletion genome is enriched, OXPHOS declines, resulting in cellular dysfunction. For example, Kearns-Sayre syndrome is caused by a single heteroplasmic mtDNA deletion. More broadly, mtDNA deletion accumulation has been observed in individual muscle cells(1) and dopaminergic neurons(2) during ageing. It is unclear how mtDNA deletions are tolerated or how they are propagated in somatic cells. One mechanism by which cells respond to OXPHOS dysfunction is by activating the mitochondrial unfolded protein response (UPRmt), a transcriptional response mediated by the transcription factor ATFS-1 that promotes the recovery and regeneration of defective mitochondria(3,4). Here we investigate the role of ATFS-1 in the maintenance and propagation of a deleterious mtDNA in a heteroplasmic Caenorhabditis elegans strain that stably expresses wild-type mtDNA and mtDNA with a 3.1-kilobase deletion (Delta mtDNA) lacking four essential genes(5). The heteroplasmic strain, which has 60% Delta mtDNA, displays modest mitochondrial dysfunction and constitutive UPRmt activation. ATFS-1 impairment reduced the Delta mtDNA nearly tenfold, decreasing the total percentage to 7%. We propose that in the context of mtDNA heteroplasmy, UPRmt activation caused by OXPHOS defects propagates or maintains the deleterious mtDNA in an attempt to recover OXPHOS activity by promoting mitochondrial biogenesis and dynamics.

More than half of the >5000 approved mineral species are known from five or fewer localities and thus are rare. Mineralogical rarity arises from different circumstances, but all rare mineral species conform to one or more of four criteria: (1) P-T-Xrange: minerals that form only under highly restricted conditions in pressure-temperature-composition space; (2) Planetary constraints: minerals that incorporate essential elements that are rare or that form at extreme conditions that seldom occur in Earth's near-surface environment; (3) Ephemeral phases: minerals that rapidly break down under ambient conditions; and (4) Collection biases: phases that are difficult to recognize because they lack crystal faces or are microscopic, or minerals that arise in lithological contexts that are difficult to access. Minerals that conform to criterion 1, 2, or 3 are inherently rare, whereas those matching criterion 4 may be much more common than represented by reported occurrences. Rare minerals, though playing minimal roles in Earth's bulk properties and dynamics, are nevertheless of significance for varied reasons. Uncommon minerals are key to understanding the diversity and disparity of Earth's mineralogical environments, for example in the prediction of as yet undescribed minerals. Novel minerals often point to extreme compositional regimes that can arise in Earth's shallow crust and they are thus critical to understanding Earth as a complex evolving system. Many rare minerals have unique crystal structures or reveal the crystal chemical plasticity of well-known structures, as dramatically illustrated by the minerals of boron. Uncommon minerals may have played essential roles in life's origins; conversely, many rare minerals arise only as a consequence, whether direct or indirect, of biological processes. The distribution of rare minerals may thus be a robust biosignature, while these phases individually and collectively exemplify the co-evolution of the geosphere and biosphere. Finally, mineralogical rarities, as with novelty in other natural domains, are inherently fascinating.

Acinetobacter baumannii is a Gram-negative bacterial pathogen responsible for a range of nosocomial infections. The recent rise and spread of multidrug-resistant A. baumannii clones has fueled a search for alternative therapies, including bacteriophage endolysins with potent antibacterial activities. A common feature of these lysins is the presence of a highly positively charged C-terminal domain with a likely role in promoting outer membrane penetration. In the present study, we show that the C-terminal amino acids 108 to 138 of phage lysin PlyF307, named P307, alone were sufficient to kill A. baumannii (> 3 logs). Furthermore, P307 could be engineered for improved activity, the most active derivative being P307(SQ-8C) (> 5-log kill). Both P307 and P307(SQ-8C) showed high in vitro activity against A. baumannii in biofilms. Moreover, P307(SQ-8C) exhibited MICs comparable to those of levofloxacin and ceftazidime and acted synergistically with polymyxin B. Although the peptides were shown to kill by disrupting the bacterial cytoplasmic membrane, they did not lyse human red blood cells or B cells; however, serum was found to be inhibitory to lytic activity. In a murine model of A. baumannii skin infection, P307(SQ-8C) reduced the bacterial burden by similar to 2 logs in 2 h. This study demonstrates the prospect of using peptide derivatives from bacteriophage lysins to treat topical infections and remove biofilms caused by Gram-negative pathogens.

Drug addiction requires learning and memory processes that are facilitated by activation of cannabinoid-1 (CB1) and opioid receptors in the hippocampus. This involves activity-dependent synaptic plasticity that is partially regulated by endogenous opioid (enkephalin and dynorphin) and non-opioid peptides, specifically cholecystokinin, parvalbumin and neuropeptide Y, the neuropeptides present in inhibitory interneurons that co-express CB1 or selective opioid receptors. We tested the hypothesis that CB1 receptor expression is a determinant of the availability of one or more of these peptide modulators in the hippocampus. This was achieved by quantitatively analyzing the immunoperoxidase labeling for each of these neuropeptide in the dorsal hippocampus of female wild-type (CB1+/+) and cannabinoid receptor 1 knockout (CB1-/-) C57/BL6 mice. The levels of Leu(5)-enkephalin-immunoreactivity were significantly reduced in the hilus of the dentate gyrus and in stratum lucidum of CA3 in CB1-/- mice. Moreover, the numbers of neuropeptide Y-immunoreactive interneurons in the dentate hilus were significantly lower in the CB1-/- compared to wild-type mice. However, CB1+/+ and CB1-/- mice did not significantly differ in expression levels of either dynorphin or cholecystokinin, and showed no differences in numbers of parvalbumin-containing interneurons. These findings suggest that the cannabinoid and opioid systems have a nuanced, regulatory relationship that could affect the balance of excitation and inhibition in the hippocampus and thus processes such as learning that rely on this balance. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

Background: Tofacitinib is an oral Janus kinase inhibitor being investigated for psoriasis. Objectives:We sought to report longer-term tofacitinib efficacy and safety in patients with moderate to severe psoriasis. Methods: Data from 2 identical phase-III studies, Oral-treatment Psoriasis Trial Pivotal 1 and 2, were pooled with data from these patients in an ongoing open-label long-term extension study. Patients (n = 1861) were randomized 2:2:1 to tofacitinib 5 mg, 10 mg, or placebo twice daily (BID). At week 16, placebo patients were rerandomized to tofacitinib. Pivotal study participants could enroll into the long-term extension where they received tofacitinib at 10 mg BID for 3 months, after which dosing could be 5 or 10 mg BID. Results: At week 28, the proportions of patients randomized to tofacitinib 5 and 10 mg BID achieving 75% or greater reduction in Psoriasis Area and Severity Index score from baseline were 55.6% and 68.8%, and achieving Physician Global Assessment of clear or almost clear were 54.7% and 65.9%. Efficacy was maintained in most patients through 24 months. Serious adverse events and discontinuations because of adverse events were reported in less than 11% of patients over 33 months of tofacitinib exposure. Limitations: There was no dose comparison beyond week 52. Conclusions: Oral tofacitinib demonstrated sustained efficacy in patients with psoriasis through 2 years, with 10 mg BID providing greater efficacy than 5 mg BID. No unexpected safety findings were observed.

The bacterial CRISPR/Cas9 system allows sequence-specific gene editing in many organisms and holds promise as a tool to generate models of human diseases, for example, in human pluripotent stem cells(1,2). CRISPR/Cas9 introduces targeted double-stranded breaks (DSBs) with high efficiency, which are typically repaired by non-homologous end-joining (NHEJ) resulting in nonspecific insertions, deletions or other mutations (indels)(2). DSBs may also be repaired by homology-directed repair (HDR)(1,2) using a DNA repair template, such as an introduced single-stranded oligo DNA nucleotide (ssODN), allowing knock-in of specific mutations(3). Although CRISPR/Cas9 is used extensively to engineer gene knockouts through NHEJ, editing by HDR remains inefficient(3-8) and can be corrupted by additional indels(9), preventing its widespread use for modelling genetic disorders through introducing disease-associated mutations. Furthermore, targeted mutational knock-in at single alleles to model diseases caused by heterozygous mutations has not been reported. Here we describe a CRISPR/Cas9-based genome-editing framework that allows selective introduction of mono-and bi-allelic sequence changes with high efficiency and accuracy. We show that HDR accuracy is increased dramatically by incorporating silent CRISPR/Casblocking mutations along with pathogenic mutations, and establish a method termed 'CORRECT' for scarless genome editing. By characterizing and exploiting a stereotyped inverse relationship between a mutation's incorporation rate and its distance to the DSB, we achieve predictable control of zygosity. Homozygous introduction requires a guide RNA targeting close to the intended mutation, whereas heterozygous introduction can be accomplished by distance-dependent suboptimal mutation incorporation or by use of mixed repair templates. Using this approach, we generated human induced pluripotent stem cells with heterozygous and homozygous dominant early onset Alzheimer's disease-causing mutations in amyloid precursor protein (APP(Swe))(10) and presenilin 1 (PSEN1M146V)(11) and derived cortical neurons, which displayed genotype-dependent disease-associated phenotypes. Our findings enable efficient introduction of specific sequence changes with CRISPR/Cas9, facilitating study of human disease.

Embryonic exposure to ethanol is known to affect neurochemical systems in rodents and increase alcohol drinking and related behaviors in humans and rodents. With zebrafish emerging as a powerful tool for uncovering neural mechanisms of numerous diseases and exhibiting similarities to rodents, the present report building,on our rat studies examined in zebrafish the effects of embryonic ethanol exposure on hypothalamic neurogenesis, expression of orexigenic neuropeptides, and voluntary ethanol consumption and locomotor behaviors in larval and adult zebrafish, and also effects of central neuropeptide injections on these behaviors affected by ethanol. At 24 h post-fertilization, zebrafish embryos were exposed for 2 h to ethanol, at low concentrations of 0.25% and 0.5%, in the tank water. Embryonic ethanol compared to control dose-dependently increased hypothalamic neurogenesis and the proliferation and expression of the orexigenic peptides, galanin (GAL) and orexin (OX), in the anterior hypothalamus. These changes in hypothalamic peptide neurons were accompanied by an increase in voluntary consumption of 10% ethanol-gelatin and in novelty-induced locomotor and exploratory behavior in adult zebrafish and locomotor activity in larvae. After intracerebroventricular injection, these peptides compared to vehicle had specific effects on these behaviors altered by ethanol, with GAL stimulating consumption of 10% ethanol-gelatin more than plain gelatin food and OX stimulating novelty-induced locomotor behavior while increasing intake of food and ethanol equally. These results, similar to those obtained in rats, suggest that the ethanol-induced increase in genesis and expression of these hypothalamic peptide neurons contribute to the behavioral changes induced by embryonic exposure to ethanol. (C) 2016 Elsevier B.V. All rights reserved.

Molecular evolution is an established technique for inferring gene homology but regulatory DNA turns over so rapidly that inference of ancestral networks is often impossible. In silico evolution is used to compute the most parsimonious path in regulatory space for anteriorposterior patterning linking two Dipterian species. The expression pattern of gap genes has evolved between Drosophila (fly) and Anopheles (mosquito), yet one of their targets, eve, has remained invariant. Our model predicts that stripe 5 in fly disappears and a new posterior stripe is created in mosquito, thus eve stripe modules 3+7 and 4+6 in fly are homologous to 3+6 and 4+5 in mosquito. We can place Clogmia on this evolutionary pathway and it shares the mosquito homologies. To account for the evolution of the other pair-rule genes in the posterior we have to assume that the ancestral Dipterian utilized a dynamic method to phase those genes in relation to eve.

Background: Transforming growth factor-beta 1 (TGF-beta 1) has been implicated in the pathogenesis of aortic valve stenosis (AS). There is, however, little direct evidence for a role of active TGF-beta 1 in AS due to the sensitivity of current assays. We searched for evidence of plasma TGF-beta 1 activation by assaying Smad2/3 phosphorylation in circulating leukocytes and platelet-leukocyte aggregates (PLAs) in a mouse model of AS (Reversa). Methods: Echocardiography was used to measure AS and cardiac function. Intracellular phospho-flow cytometry in combination with optical fluorescence microscopy was used to detect PLAs and p-Smad2/3 levels. Results: Reversa mice on a western diet developed AS, had significantly increased numbers of PLAs and more intense staining for p-Smad2/3 in both PLAs and single leukocytes (all p < 0.05). p-Smad2/3 staining was more intense in PLAs than in single leukocytes in both diet groups (p < 0.05) and correlated with plasma total TGF-beta 1 levels (r = 0.38, p = 0.05 for PLAs and r = 0.37, p = 0.06 for single leukocytes) and reductions in ejection fraction (r = 0.42, p = 0.03 for PLAs and r = 037, p = 0.06 for single leukocytes). Conclusions: p-Smad2/3 staining is more intense in leukocytes of hypercholesterolemic mice that developed AS, suggesting increased circulating active TGF-beta 1 levels. Leukocyte p-Smad2/3 may be a valuable surrogate indicator of circulating active TGF-beta 1. (C) 2016 Elsevier Inc All rights reserved.

A measurement of the ratio of the branching fractions of the B-s(0) meson to J/psi f(0)(980) and to J/psi phi(1020) is presented. The J/psi, f(0)(980), and phi(1020) are observed through their decays to mu(+)mu(-), pi(+)pi(-), and K+K-, respectively. The f(0) and the phi are identified by requiring |M-pi+(pi)- - 974 MeV| < 50 MeV and |M-K+(K)- - 1020 MeV| < 10 MeV. The analysis is based on a data sample of pp collisions at a centre-of-mass energy of 7 TeV, collected by the CMS experiment at the LHC, corresponding to an integrated luminosity of 5.3fb(-1). The measured ratio is B(B-s(0) -> J/psi f(0)) B(f(0) ->pi(+)pi(-))/B(B-s(0) -> J/psi phi) B(phi -> K+K-)= 0.140 +/- 0.008 (stat) +/- 0.023 (syst), where the first uncertainty is statistical and the second is systematic. (C) 2016 CERN for the benefit of the CMS Collaboration. Published by Elsevier B.V. This is an open access article under the CC BY license.