bluuebunny/crossref_metadata_2025 huggingface dataset:
Susceptibility of Human Endogenous Retrovirus Type K to Reverse Transcriptase Inhibitors
Authors: Contreras-Galindo Rafael , Dube Derek , Fujinaga Koh , Kaplan Mark H. , Markovitz David M.
Published: September 2017

Human endogenous retroviruses (HERVs) make up 8% of the human genome. The HERV type K (HERV-K) HML-2 (HK2) family contains proviruses that are the most recent entrants into the human germ line and are transcriptionally active. In HIV-1 infection and cancer, HK2 genes produce retroviral particles that appear to be infectious, yet the replication capacity of these viruses and potential pathogenicity has been difficult to ascertain. In this report, we screened the efficacy of commercially available reverse transcriptase inhibitors (RTIs) at inhibiting the enzymatic activity of HK2 RT and HK2 genomic replication. Interestingly, only one provirus, K103, was found to encode a functional RT among those examined. Several nucleoside analogue RTIs (NRTIs) blocked K103 RT activity and consistently inhibited the replication of HK2 genomes. The NRTIs zidovudine (AZT), stavudine (d4T), didanosine (ddI), and lamivudine (3TC), and the nucleotide RTI inhibitor tenofovir (TDF), show efficacy in blocking K103 RT. HIV-1-specific nonnucleoside RTIs (NNRTIs), protease inhibitors (PIs), and integrase inhibitors (IIs) did not affect HK2, except for the NNRTI etravirine (ETV). The inhibition of HK2 infectivity by NRTIs appears to take place at either the reverse transcription step of the viral genome prior to HK2 viral particle formation and/or in the infected cells. Inhibition of HK2 by these drugs will be useful in suppressing HK2 infectivity if these viruses prove to be pathogenic in cancer, neurological disorders, or other diseases associated with HK2. The present studies also elucidate a key aspect of the life cycle of HK2, specifically addressing how they do, and/or did, replicate.

IMPORTANCEEndogenous retroviruses are relics of ancestral virus infections in the human genome. The most recent of these infections was caused by HK2. While HK2 often remains silent in the genome, this group of viruses is activated in HIV-1-infected and cancer cells. Recent evidence suggests that these viruses are infectious, and the potential exists for HK2 to contribute to disease. We show that HK2, and specifically the enzyme that mediates virus replication, can be inhibited by a panel of drugs that are commercially available. We show that several drugs block HK2 with different efficacies. The inhibition of HK2 replication by antiretroviral drugs appears to occur in the virus itself as well as after infection of cells. Therefore, these drugs might prove to be an effective treatment by suppressing HK2 infectivity in diseases where these viruses have been implicated, such as cancer and neurological syndromes.
p21 Restricts HIV-1 in Monocyte-Derived Dendritic Cells through the Reduction of Deoxynucleoside Triphosphate Biosynthesis and Regulation of SAMHD1 Antiviral Activity
Authors: Valle-Casuso Jose Carlos , Allouch Awatef , David Annie , Lenzi Gina M. , Studdard Lydia , Barré-Sinoussi Françoise , Müller-Trutwin Michaela , Kim Baek , Pancino Gianfranco , Sáez-Cirión Asier
Published: September 2017

HIV-1 infection of noncycling cells, such as dendritic cells (DCs), is impaired due to limited availability of deoxynucleoside triphosphates (dNTPs), which are needed for HIV-1 reverse transcription. The levels of dNTPs are tightly regulated during the cell cycle and depend on the balance between dNTP biosynthesis and degradation. SAMHD1 potently blocks HIV-1 replication in DCs, although the underlying mechanism is still unclear. SAMHD1 has been reported to be able to degrade dNTPs and viral nucleic acids, which may both hamper HIV-1 reverse transcription. The relative contribution of these activities may differ in cycling and noncycling cells. Here, we show that inhibition of HIV-1 replication in monocyte-derived DCs (MDDCs) is associated with an increased expression of p21cip1/waf, a cell cycle regulator that is involved in the differentiation and maturation of DCs. Induction of p21 in MDDCs decreases the pool of dNTPs and increases the antiviral active isoform of SAMHD1. Although both processes are complementary in inhibiting HIV-1 replication, the antiviral activity of SAMHD1 in our primary cell model appears to be, at least partially, independent of its dNTPase activity. The reduction in the pool of dNTPs in MDDCs appears rather mostly due to a p21-mediated suppression of several enzymes involved in dNTP synthesis (i.e., RNR2, TYMS, and TK-1). These results are important to better understand the interplay between HIV-1 and DCs and may inform the design of new therapeutic approaches to decrease viral dissemination and improve immune responses against HIV-1.

IMPORTANCE
DCs play a key role in the induction of immune responses against HIV. However, HIV has evolved ways to exploit these cells, facilitating immune evasion and virus dissemination. We have found that the expression of p21, a cyclin-dependent kinase inhibitor involved in cell cycle regulation and monocyte differentiation and maturation, potentially can contribute to the inhibition of HIV-1 replication in monocyte-derived DCs through multiple mechanisms. p21 decreased the size of the intracellular dNTP pool. In parallel, p21 prevented SAMHD1 phosphorylation and promoted SAMHD1 dNTPase-independent antiviral activity. Thus, induction of p21 resulted in conditions that allowed the effective inhibition of HIV-1 replication through complementary mechanisms. Overall, p21 appears to be a key regulator of HIV infection in myeloid cells.
A DNA Vaccine That Targets Hemagglutinin to Antigen-Presenting Cells Protects Mice against H7 Influenza
Authors: Andersen Tor Kristian , Zhou Fan , Cox Rebecca , Bogen Bjarne , Grødeland Gunnveig
Published: September 2017

Zoonotic influenza H7 viral infections have a case fatality rate of about 40%. Currently, no or limited human to human spread has occurred, but we may be facing a severe pandemic threat if the virus acquires the ability to transmit between humans. Novel vaccines that can be rapidly produced for global distribution are urgently needed, and DNA vaccines may be the only type of vaccine that allows for the speed necessary to quench an emerging pandemic. Here, we constructed DNA vaccines encoding the hemagglutinin (HA) from influenza A/chicken/Italy/13474/99 (H7N1). In order to increase the efficacy of DNA vaccination, HA was targeted to either major histocompatibility complex class II molecules or chemokine receptors 1, 3, and 5 (CCR1/3/5) that are expressed on antigen-presenting cells (APC). A single DNA vaccination with APC-targeted HA significantly increased antibody levels in sera compared to nontargeted control vaccines. The antibodies were confirmed neutralizing in an H7 pseudotype-based neutralization assay. Furthermore, the APC-targeted vaccines increased the levels of antigen-specific cytotoxic T cells, and a single DNA vaccination could confer protection against a lethal challenge with influenza A/turkey/Italy/3889/1999 (H7N1) in mice. In conclusion, we have developed a vaccine that rapidly could contribute protection against a pandemic threat from avian influenza.

IMPORTANCEHighly pathogenic avian influenza H7 constitute a pandemic threat that can cause severe illness and death in infected individuals. Vaccination is the main method of prophylaxis against influenza, but current vaccine strategies fall short in a pandemic situation due to a prolonged production time and insufficient production capabilities. In contrast, a DNA vaccine can be rapidly produced and deployed to prevent the potential escalation of a highly pathogenic influenza pandemic. We here demonstrate that a single DNA delivery of hemagglutinin from an H7 influenza could mediate full protection against a lethal challenge with H7N1 influenza in mice. Vaccine efficacy was contingent on targeting of the secreted vaccine protein to antigen-presenting cells.
Zika Virus Encoding Nonglycosylated Envelope Protein Is Attenuated and Defective in Neuroinvasion
Authors: Annamalai Arun S. , Pattnaik Aryamav , Sahoo Bikash R. , Muthukrishnan Ezhumalai , Natarajan Sathish Kumar , Steffen David , Vu Hiep L. X. , Delhon Gustavo , Osorio Fernando A. , Petro Thomas M. , Xiang Shi-Hua , Pattnaik Asit K.
Published: September 2017

Zika virus (ZIKV), a mosquito-transmitted flavivirus responsible for sporadic outbreaks of mild and febrile illness in Africa and Asia, reemerged in the last decade causing serious human diseases, including microcephaly, congenital malformations, and Guillain-Barré syndrome. Although genomic and phylogenetic analyses suggest that genetic evolution may have led to the enhanced virulence of ZIKV, experimental evidence supporting the role of specific genetic changes in virulence is currently lacking. One sequence motif, VNDT, containing an N-linked glycosylation site in the envelope (E) protein, is polymorphic; it is absent in many of the African isolates but present in all isolates from the recent outbreaks. In the present study, we investigated the roles of this sequence motif and glycosylation of the E protein in the pathogenicity of ZIKV. We first constructed a stable full-length cDNA clone of ZIKV in a novel linear vector from which infectious virus was recovered. The recombinant ZIKV generated from the infectious clone, which contains the VNDT motif, is highly pathogenic and causes lethality in a mouse model. In contrast, recombinant viruses from which the VNDT motif is deleted or in which the N-linked glycosylation site is mutated by single-amino-acid substitution are highly attenuated and nonlethal. The mutant viruses replicate poorly in the brains of infected mice when inoculated subcutaneously but replicate well following intracranial inoculation. Our findings provide the first evidence that N-linked glycosylation of the E protein is an important determinant of ZIKV virulence and neuroinvasion.

IMPORTANCE
The recent emergence of Zika virus (ZIKV) in the Americas has caused major worldwide public health concern. The virus appears to have gained significant pathogenicity, causing serious human diseases, including microcephaly and Guillain-Barré syndrome. The factors responsible for the emergence of pathogenic ZIKV are not understood at this time, although genetic changes have been shown to facilitate virus transmission. All isolates from the recent outbreaks contain an N-linked glycosylation site within the viral envelope (E) protein, whereas many isolates of the African lineage virus lack this site. To elucidate the functional significance of glycosylation in ZIKV pathogenicity, recombinant ZIKVs from infectious clones with or without the glycan on the E protein were generated. ZIKVs lacking the glycan were highly attenuated for the ability to cause mortality in a mouse model and were severely compromised for neuroinvasion. Our studies suggest glycosylation of the E protein is an important factor contributing to ZIKV pathogenicity.
Drug-Free Approach To Study the Unusual Cell Cycle of Giardia intestinalis
Authors: Horlock-Roberts Kathleen , Reaume Chase , Dayer Guillem , Ouellet Christine , Cook Nicholas , Yee Janet
Published: September 2017

Giardias are among the most commonly reported intestinal protozoa in the world, with infections seen in humans and over 40 species of animals. The life cycle of giardia alternates between the motile trophozoite and the infectious cyst. The regulation of the cell cycle controls the proliferation of giardia trophozoites during an active infection and contains the restriction point for the differentiation of trophozoite to cyst. Here, we developed counterflow centrifugal elutriation as a drug-free method to obtain fractions of giardia cultures enriched in cells from the G
1
, S, and G
2
stages of the cell cycle. Analysis of these fractions showed that the cells do not show side effects associated with the drugs used for synchronization of giardia cultures. Therefore, counterflow centrifugal elutriation would advance studies on key regulatory events during the giardia cell cycle and identify potential drug targets to block giardia proliferation and transmission.
Zebrafish (Danio rerio) as a Vertebrate Model Host To Study Colonization, Pathogenesis, and Transmission of Foodborne Escherichia coli O157
Authors: Stones Daniel H. , Fehr Alexander G. J. , Thompson Laurel , Rocha Jacqueline , Perez-Soto Nicolas , Madhavan Vipin T. P. , Voelz Kerstin , Krachler Anne Marie
Published: September 2017

Enterohemorrhagic
Escherichia coli
(EHEC) is a foodborne pathogen which can cause diarrhea, vomiting, and, in some cases, severe complications such as kidney failure in humans. Up to 30% of cattle are colonized with EHEC, which can enter the food chain through contaminated meat, dairy, and vegetables. In order to control infections and stop transmission, it is important to understand what factors allow EHEC to colonize its hosts, cause virulence, and aid transmission. Since this cannot be systematically studied in humans, it is important to develop animal models of infection and transmission. We developed a model which allows us to study foodborne infection in zebrafish, a vertebrate host that is transparent and genetically tractable. Our results show that foodborne infection is more efficient than waterborne infection and that the locus of enterocyte effacement is a key virulence determinant in the zebrafish model. It is induced early during infection, and loss of tight LEE regulation leads to a decreased bacterial burden and decreased host mortality. Overall, the zebrafish model allows us to study foodborne infection, including pathogen release from the food vehicle and gene regulation and its context of host-microbe interactions, as well as environmental shedding and transmission to naive hosts.
On the foundations of general relativistic celestial mechanics
Authors: Battista Emmanuele , Esposito Giampiero , Dell’Agnello Simone
Published: September 2017

Towards the end of nineteenth century, Celestial Mechanics provided the most powerful tools to test Newtonian gravity in the solar system and also led to the discovery of chaos in modern science. Nowadays, in light of general relativity, Celestial Mechanics leads to a new perspective on the motion of satellites and planets. The reader is here introduced to the modern formulation of the problem of motion, following what the leaders in the field have been teaching since the nineties, in particular, the use of a global chart for the overall dynamics of N bodies and N local charts describing the internal dynamics of each body. The next logical step studies in detail how to split the N-body problem into two sub-problems concerning the internal and external dynamics, how to achieve the effacement properties that would allow a decoupling of the two sub-problems, how to define external-potential-effacing coordinates and how to generalize the Newtonian multipole and tidal moments. The review paper ends with an assessment of the nonlocal equations of motion obtained within such a framework, a description of the modifications induced by general relativity on the theoretical analysis of the Newtonian three-body problem, and a mention of the potentialities of the analysis of solar-system metric data carried out with the Planetary Ephemeris Program.
Supermultiplet of β-deformations from twistors
Authors: Milián Segundo P.
Published: September 2017

We consider the supermultiplet of linearized beta-deformation of [Formula: see text] super-Yang–Mills (SYM). It was previously studied on the gravitational side. We study the supermultiplet of beta-deformations on the field theory side and we compare two finite-dimensional representations of [Formula: see text] algebra. We show that they are related by an intertwining operator. We develop a twistor-based approach which could be useful for studying other finite-dimensional and nonunitary representations in AdS/CFT correspondence.
A Clifford algebra approach to chiral symmetry breaking and fermion mass hierarchies
Authors: Lu Wei
Published: September 2017

We propose a Clifford algebra approach to chiral symmetry breaking and fermion mass hierarchies in the context of composite Higgs bosons. Standard model fermions are represented by algebraic spinors of six-dimensional binary Clifford algebra, while ternary Clifford algebra-related flavor projection operators control allowable flavor-mixing interactions. There are three composite electroweak Higgs bosons resulted from top quark, tau neutrino, and tau lepton condensations. Each of the three condensations gives rise to masses of four different fermions. The fermion mass hierarchies within these three groups are determined by four-fermion condensations, which break two global chiral symmetries. The four-fermion condensations induce axion-like pseudo-Nambu–Goldstone bosons and can be dark matter candidates. In addition to the 125 GeV Higgs boson observed at the Large Hadron Collider, we anticipate detection of tau neutrino composite Higgs boson via the charm quark decay channel.
Ultrathin Au film on polymer surface for surface plasmon polariton waveguide application
Authors: Liu Tong , Ji Lanting , He Guobing , Sun Xiaoqiang , Wang Fei , Zhang Daming
Published: September 2017

Formation of laterally continuous ultrathin gold films on polymer substrates is a technological challenge. In this work, the vacuum thermal evaporation method is adopted to form continuous Au films in the thickness range of 7–17 nm on polymers of Poly(methyl-methacrylate-glycidly-methacrylate) and SU-8 film surface without using the adhesion or metallic seeding layers. Absorption spectrum, scanning electron microscope and atomic force microscope images are used to characterize the Au film thickness, roughness and optical loss. The result shows that molecular-scale structure, surface energy and electronegativity have impacts on the Au film morphology on polymers. Wet chemical etching is used to fabricate 7-nm thick Au stripes embedded in polymer claddings. These long-range surface plasmon polariton waveguides demonstrate the favorable morphological configurations and cross-sectional states. Through the end-fire excitation method, propagation losses of 6-[Formula: see text] wide Au stripes are compared to theoretical values and analyzed from practical film status. The smooth, patternable gold films on polymer provide potential applications to plasmonic waveguides, biosensing, metamaterials and optical antennas.
Magnetic fields in turbulent quark matter and magnetar bursts
Authors: Dvornikov Maxim
Published: September 2017

We analyze the magnetic field evolution in dense quark matter with unbroken chiral symmetry, which can be found inside quark and hybrid stars. The magnetic field evolves owing to the chiral magnetic effect in the presence of the electroweak interaction between quarks. In our study, we also take into account the magnetohydrodynamic turbulence effects in dense quark matter. We derive the kinetic equations for the spectra of the magnetic helicity density and the magnetic energy density as well as for the chiral imbalances. On the basis of the numerical solution of these equations, we find that turbulence effects are important for the behavior of small scale magnetic fields. It is revealed that, under certain initial conditions, these magnetic fields behave similarly to the electromagnetic flashes of some magnetars. We suggest that fluctuations of magnetic fields, described in frames of our model, which are created in the central regions of a magnetized compact star, can initiate magnetar bursts.
Synthesis of CeO2 Porous Nanospheres with Tunable Pore and Particle Sizes and Their Adsorption to Methyl Orange
Authors: Li Longfeng , Li Chengyu , Liu Mingzhu , Wang Cheng
Published: September 2017

The catalytic performance of ceria oxide (CeO[Formula: see text] was greatly influenced by the materials’ structure and surface area. In this paper, CeO2 porous nanospheres (PNS) with tunable pore and sphere sizes were synthesized by a solvothermal method using diethylene glycol as reaction media. All the as-synthesized CeO2 PNS presented good adsorption capacity for methyl orange, which makes them potential application in waste water treatment conveniently and economically. The control over the sphere and pore sizes of CeO2 PNS was achieved by regulating the volume of water bubble in the reaction system, which supplied limited room for the assembling of preformed CeO2 small nano-precursors.
Identifying Causes of Patterns in Ecological Networks: Opportunities and Limitations
Authors: Dormann Carsten F. , Fründ Jochen , Schaefer H. Martin
Published: September 2017

Ecological networks depict the interactions between species, mainly based on observations in the field. The information contained in such interaction matrices depends on the sampling design, and typically, compounds preferences (specialization) and abundances (activity). Null models are the primary vehicles to disentangle the effects of specialization from those of sampling and abundance, but they ignore the feedback of network structure on abundances. Hence, network structure, as exemplified here by modularity, is difficult to link to specific causes. Indeed, various processes lead to modularity and to specific interaction patterns more generally. Inferring (co)evolutionary dynamics is even more challenging, as competition and trait matching yield identical patterns of interactions. A satisfactory resolution of the underlying factors determining network structure will require substantial additional information, not only on independently assessed abundances, but also on traits, and ideally on fitness consequences as measured in experimental setups.
Combining Traditional Mutagenesis with New High-Throughput Sequencing and Genome Editing to Reveal Hidden Variation in Polyploid Wheat
Authors: Uauy Cristobal , Wulff Brande B.H. , Dubcovsky Jorge
Published: September 2017

Induced mutations have been used to generate novel variation for breeding purposes since the early 1900s. However, the combination of this old technology with the new capabilities of high-throughput sequencing has resulted in powerful reverse genetic approaches in polyploid crops. Sequencing genomes or exomes of large mutant populations can generate extensive databases of mutations for most genes. These mutant collections, together with genome editing, are being used in polyploid species to combine mutations in all copies of a gene (homoeologs), and to expose phenotypic variation that was previously hidden by functional redundancy among homoeologs. This redundancy is more extensive in recently formed polyploids such as wheat, which can now benefit from the deployment of useful recessive mutations previously identified in its diploid relatives. Sequenced mutant populations and genome editing have changed the paradigm of what is possible in functional genetic analysis of wheat.
Game-Theoretic Approaches to Pragmatics
Authors: Benz Anton , Stevens Jon
Published: September 2017

We present an overview and comparison of different game-theoretic approaches to Gricean pragmatics, including games of partial information, optimal answer models, error models, iterated best response models, and rational speech act models. We address phenomena of disambiguation, scalar implicature, and relevance implicature.
A Dynamic, Inclusive, and Affective Evolutionary View of Organizational Behavior
Authors: Mitchell Terence R.
Published: September 2017

This article reflects my observations about major changes in organizational behavior (OB) during my 50 years working in the field. I begin with a brief description of my personal journey as foundational for my perspective. I then discuss how the field has changed structurally and focus on the contributions of the Academy of Management (AoM) and the increasing diversity of our members, especially the increasing number of women, minorities, and people from countries other than the United States. The next section covers major research content changes in the field including positive scholarship, leadership, affective constructs, ethics, and the general well-being of those who toil in the organizational trenches. Additionally, there is a section on how the utilization of new statistical procedures and research methods has greatly expanded our ability to investigate and test complex theories and hypotheses about behavior in organizations. The discussion section highlights both the positive aspects of these developments along with some concerns about the future.
Hemophagocytic Lymphohistiocytosis
Authors: Al-Samkari Hanny , Berliner Nancy
Published: September 2017

Hemophagocytic lymphohistiocytosis is a life-threatening disorder characterized by unbridled activation of cytotoxic T lymphocytes, natural killer (NK) cells, and macrophages resulting in hypercytokinemia and immune-mediated injury of multiple organ systems. It is seen in both children and adults and is recognized as primary (driven by underlying genetic mutations that abolish critical proteins required for normal function of cytotoxic T cells and NK cells) or secondary (resulting from a malignant, infectious, or autoimmune stimulus without an identifiable underlying genetic trigger). Clinical and laboratory manifestations include fever, splenomegaly, neurologic dysfunction, coagulopathy, liver dysfunction, cytopenias, hypertriglyceridemia, hyperferritinemia, hemophagocytosis, and diminished NK cell activity. It is treated with immune suppressants, etoposide, and allogeneic hematopoietic stem cell transplantation; more than 50% of children who undergo transplant survive, but adults have quite poor outcomes even with aggressive management. Newer agents directed at subduing the uncontrolled immune response in a targeted fashion offer promise in this highly morbid disease.
Modulators of actin-myosin dissociation: basis for muscle type functional differences during fatigue
Authors: Karatzaferi Christina , Adamek Nancy , Geeves Michael A.
Published: September 2017

The muscle types present with variable fatigue tolerance, in part due to the myosin isoform expressed. However, the critical steps that define “fatigability” in vivo of fast vs. slow myosin isoforms, at the molecular level, are not yet fully understood. We examined the modulation of the ATP-induced myosin subfragment 1 (S1) dissociation from pyrene-actin by inorganic phosphate (Pi), pH, and temperature using a specially modified stopped-flow system that allowed fast kinetics measurements at physiological temperature. We contrasted the properties of rabbit psoas (fast) and bovine masseter (slow) myosins (obtained from samples collected from New Zealand rabbits and from a licensed abattoir, respectively, according to institutional and national ethics permits). To identify ATP cycling biochemical intermediates, we assessed ATP binding to a preequilibrated mixture of actomyosin and variable [ADP], pH (pH 7 vs. pH 6.2), and Pi (zero, 15, or 30 added mM Pi) in a range of temperatures (5 to 45°C). Temperature and pH variations had little, if any, effect on the ADP dissociation constant ( KADP) for fast S1, but for slow S1, KADP was weakened with increasing temperature or low pH. In the absence of ADP, the dissociation constant for phosphate ( KPi) was weakened with increasing temperature for fast S1. In the presence of ADP, myosin type differences were revealed at the apparent phosphate affinity, depending on pH and temperature. Overall, the newly revealed kinetic differences between myosin types could help explain the in vivo observed muscle type functional differences at rest and during fatigue.
Detachment-induced E-cadherin expression promotes 3D tumor spheroid formation but inhibits tumor formation and metastasis of lung cancer cells
Authors: Powan Phattrakorn , Luanpitpong Sudjit , He Xiaoqing , Rojanasakul Yon , Chanvorachote Pithi
Published: September 2017

The epithelial-to-mesenchymal transition is proposed to be a key mechanism responsible for metastasis-related deaths. Similarly, cancer stem cells (CSCs) have been proposed to be a key driver of tumor metastasis. However, the link between the two events and their control mechanisms is unclear. We used a three-dimensional (3D) tumor spheroid assay and other CSC-indicating assays to investigate the role of E-cadherin in CSC regulation and its association to epithelial-to-mesenchymal transition in lung cancer cells. Ectopic overexpression and knockdown of E-cadherin were found to promote and retard, respectively, the formation of tumor spheroids in vitro but had opposite effects on tumor formation and metastasis in vivo in a xenograft mouse model. We explored the discrepancy between the in vitro and in vivo results and demonstrated, for the first time, that E-cadherin is required as a component of a major survival pathway under detachment conditions. Downregulation of E-cadherin increased the stemness of lung cancer cells but had an adverse effect on their survival, particularly on non-CSCs. Such downregulation also promoted anoikis resistance and invasiveness of lung cancer cells. These results suggest that anoikis assay could be used as an alternative method for in vitro assessment of CSCs that involves dysregulated adhesion proteins. Our data also suggest that agents that restore E-cadherin expression may be used as therapeutic agents for metastatic cancers.
Molecular mechanisms involved in the adaptive regulation of the colonic thiamin pyrophosphate uptake process
Authors: Anandam Kasin Yadunandam , Srinivasan Padmanabhan , Subramanian Veedamali S. , Said Hamid M.
Published: September 2017

A considerable amount of the thiamin generated by gut microbiota exists in the form of thiamin pyrophosphate (TPP). We have previously shown that human colonocytes possess an efficient carrier-mediated uptake process for TPP that involves the SLC44A4 system and this uptake process is adaptively regulated by prevailing extracellular TPP level. Little is known about the molecular mechanisms that mediate this adaptive regulation. We addressed this issue using human-derived colonic epithelial NCM460 cells and mouse colonoids as models. Maintaining NCM460 cells in the presence of a high level of TPP (1 mM) for short (2 days)- and long-term (9 days) periods was found to lead to a significant reduction in [3H] TPP uptake compared with cells maintained in its absence. Short-term exposure showed no changes in level of expression of SLC44A4 protein in total cell homogenate (although there was a decreased expression in the membrane fraction), mRNA, and promoter activity. However, a significant reduction in the level of expression of the SLC44A4 protein, mRNA, and promoter activity was observed upon long-term maintenance with the substrate. Similar changes in Slc44a4 mRNA expression were observed when mouse colonoids were maintained with TPP for short- and long-term periods. Expression of the transcription factors ELF3 and CREB-1 (which drive the SLC44A4 promoter) following long-term exposure was unchanged, but their binding affinity to the promoter was decreased and specific histone modifications were also observed. These studies demonstrate that, depending on the period of exposure, different mechanisms are involved in the adaptive regulation of colonic TPP uptake by extracellular substrate level.
Rack1 function in intestinal epithelia: regulating crypt cell proliferation and regeneration and promoting differentiation and apoptosis
Authors: Cheng Zhuan-Fen , Pai Reetesh K. , Cartwright Christine A.
Published: September 2017

Previously, we showed that receptor for activated C kinase 1 (Rack1) regulates growth of colon cells in vitro, partly by suppressing Src kinase activity at key cell cycle checkpoints, in apoptotic and cell survival pathways and at cell-cell adhesions. Here, we generated mouse models of Rack1 deficiency to assess Rack1’s function in intestinal epithelia in vivo. Intestinal Rack1 deficiency resulted in proliferation of crypt cells, diminished differentiation of crypt cells into enterocyte, goblet, and enteroendocrine cell lineages, and expansion of Paneth cell populations. Following radiation injury, the morphology of Rack1-deleted small bowel was strikingly abnormal with development of large polypoid structures that contained many partly formed villi, numerous back-to-back elongated and regenerating crypts, and high-grade dysplasia in surface epithelia. These abnormalities were not observed in Rack1-expressing areas of intestine or in control mice. Following irradiation, apoptosis of enterocytes was strikingly reduced in Rack1-deleted epithelia. These novel findings reveal key functions for Rack1 in regulating growth of intestinal epithelia: suppressing crypt cell proliferation and regeneration, promoting differentiation and apoptosis, and repressing development of neoplasia.

NEW & NOTEWORTHY Our findings reveal novel functions for receptor for activated C kinase 1 (Rack1) in regulating growth of intestinal epithelia: suppressing crypt cell proliferation and regeneration, promoting differentiation and apoptosis, and repressing development of neoplasia.
Phagocytosis of microparticles by alveolar macrophages during acute lung injury requires MerTK
Authors: Mohning Michael P. , Thomas Stacey M. , Barthel Lea , Mould Kara J. , McCubbrey Alexandria L. , Frasch S. Courtney , Bratton Donna L. , Henson Peter M. , Janssen William J.
Published: September 2017

Microparticles are a newly recognized class of mediators in the pathophysiology of lung inflammation and injury, but little is known about the factors that regulate their accumulation and clearance. The primary objective of our study was to determine whether alveolar macrophages engulf microparticles and to elucidate the mechanisms by which this occurs. Alveolar microparticles were quantified in bronchoalveolar fluid of mice with lung injury induced by LPS and hydrochloric acid. Microparticle numbers were greatest at the peak of inflammation and declined as inflammation resolved. Isolated, fluorescently labeled particles were placed in culture with macrophages to evaluate ingestion in the presence of endocytosis inhibitors. Ingestion was blocked with cytochalasin D and wortmannin, consistent with a phagocytic process. In separate experiments, mice were treated intratracheally with labeled microparticles, and their uptake was assessed though microscopy and flow cytometry. Resident alveolar macrophages, not recruited macrophages, were the primary cell-ingesting microparticles in the alveolus during lung injury. In vitro, microparticles promoted inflammatory signaling in LPS primed epithelial cells, signifying the importance of microparticle clearance in resolving lung injury. Microparticles were found to have phosphatidylserine exposed on their surfaces. Accordingly, we measured expression of phosphatidylserine receptors on macrophages and found high expression of MerTK and Axl in the resident macrophage population. Endocytosis of microparticles was markedly reduced in MerTK-deficient macrophages in vitro and in vivo. In conclusion, microparticles are released during acute lung injury and peak in number at the height of inflammation. Resident alveolar macrophages efficiently clear these microparticles through MerTK-mediated phagocytosis.
Regulation of genes related to cholesterol metabolism in rainbow trout (Oncorhynchus mykiss) fed a plant-based diet
Authors: Zhu Tengfei , Corraze Geneviève , Plagnes-Juan Elisabeth , Quillet Edwige , Dupont-Nivet Mathilde , Skiba-Cassy Sandrine
Published: September 2017

When compared with fish meal and fish oil, plant ingredients differ not only in their protein content and amino acid and fatty acid profiles but are also devoid of cholesterol, the major component of cell membrane and precursor of several bioactive compounds. Based on these nutritional characteristics, plant-based diets can affect fish physiology and cholesterol metabolism. To investigate the mechanisms underlying cholesterol homeostasis, rainbow trout were fed from 1 g body wt for 6 mo with a totally plant-based diet (V), a marine diet (M), and a marine-restricted diet (MR), with feed intake adjusted to that of the V group. The expression of genes involved in cholesterol synthesis, esterification, excretion, bile acid synthesis, and cholesterol efflux was measured in liver. Results showed that genes involved in cholesterol synthesis were upregulated in trout fed the V diet, whereas expression of genes related to bile acid synthesis ( cyp7a1) and cholesterol elimination ( abcg8) were reduced. Feeding trout the V diet also enhanced the expression of srebp-2 while reducing that of lxrα and miR-223. Overall, these data suggested that rainbow trout coped with the altered nutritional characteristics and absence of dietary cholesterol supply by increasing cholesterol synthesis and limiting cholesterol efflux through molecular mechanisms involving at least srebp-2, lxrα, and miR-223. However, plasma and body cholesterol levels in trout fed the V diet were lower than in fish fed the M diet, raising the question of the role of cholesterol in the negative effect of plant-based diet on growth.
PGC1α in the kidney
Authors: Lynch Matthew R. , Tran Mei T. , Parikh Samir M.
Published: September 2017

Acute kidney injury (AKI) arising from diverse etiologies is characterized by mitochondrial dysfunction. The peroxisome proliferator-activated receptor γ coactivator-1alpha (PGC1α), a master regulator of mitochondrial biogenesis, has been shown to be protective in AKI. Interestingly, reduction of PGC1α has also been implicated in the development of diabetic kidney disease and renal fibrosis. The beneficial renal effects of PGC1α make it a prime target for therapeutics aimed at ameliorating AKI, forms of chronic kidney disease (CKD), and their intersection. This review summarizes the current literature on the relationship between renal health and PGC1α and proposes areas of future interest.
Three-dimensional morphometric analysis of the renal vasculature
Authors: Khan Zohaib , Ngo Jennifer P. , Le Bianca , Evans Roger G. , Pearson James T. , Gardiner Bruce S. , Smith David W.
Published: September 2017

Vascular topology and morphology are critical in the regulation of blood flow and the transport of small solutes, including oxygen, carbon dioxide, nitric oxide, and hydrogen sulfide. Renal vascular morphology is particularly challenging, since many arterial walls are partially wrapped by the walls of veins. In the absence of a precise characterization of three-dimensional branching vascular geometry, accurate computational modeling of the intrarenal transport of small diffusible molecules is impossible. An enormous manual effort was required to achieve a relatively precise characterization of rat renal vascular geometry, highlighting the need for an automated method for analysis of branched vasculature morphology to allow characterization of the renal vascular geometry of other species, including humans. We present a semisupervised method for three-dimensional morphometric analysis of renal vasculature images generated by computed tomography. We derive quantitative vascular attributes important to mass transport between arteries, veins, and the renal tissue and present methods for their computation for a three-dimensional vascular geometry. To validate the algorithm, we compare automated vascular estimates with subjective manual measurements for a portion of rabbit kidney. Although increased image resolution can improve outcomes, our results demonstrate that the method can quantify the morphological characteristics of artery-vein pairs, comparing favorably with manual measurements. Similar to the rat, we show that rabbit artery-vein pairs become less intimate along the course of the renal vasculature, but the total wrapped mass transfer coefficient increases and then decreases. This new method will facilitate new quantitative physiological models describing the transport of small molecules within the kidney.
Effects of expiratory muscle activation via high-frequency spinal cord stimulation
Authors: Kowalski K. E. , Romaniuk J. R. , Kowalski T. , DiMarco A. F.
Published: September 2017

In persons with spinal cord injury, lower thoracic low-frequency spinal cord stimulation (LF-SCS; 50 Hz, 15 mA) is a useful method to restore an effective cough. Unfortunately, the high-stimulus-amplitude requirements and potential activation of pain fibers significantly limit this application in persons with intact sensation. In this study, the mechanism of the expiratory muscle activation, via high-frequency SCS (HF-SCS; 500 Hz, 1 mA) was evaluated in dogs. In group 1, the effects of electrode placement on airway pressure generation (P) was evaluated. Maximal P occurred at the T9–T10 level with progressive decrements in P at more rostral and caudal levels for both LF-SCS and HF-SCS. In group 2, electromyographic (EMG) latencies of internal intercostal muscle (II) activation were evaluated before and after spinal root section and during direct motor root stimulation. Onset time of II EMG activity during HF-SCS was significantly longer (3.84 ± 1.16 ms) than obtained during direct motor root activation (1.61 ± 0.10 ms). In group 3, P and external oblique (EO) EMG activity, before and after sequential spinal section at the T11–T12 level, were evaluated. Bilateral dorsal column section significantly reduced EO EMG activity below the section and resulted in a substantial fall in P. Subsequent lateral funiculi section completely abolished those activities and resulted in further reductions in P. We conclude that 1) activation of the expiratory muscles via HF-SCS is dependent entirely on synaptic spinal cord pathways, and 2) HF-SCS at the T9 level produces a comparable level of muscle activation with that achieved with LF-SCS but with much lower stimulus amplitudes.

NEW & NOTEWORTHY The findings in the present study suggest that lower thoracic high-frequency spinal cord stimulation with low stimulus currents results in sufficient activation of the expiratory muscles via spinal circuitry to produce large positive airway pressures sufficient to generate an effective cough mechanism. This method, therefore, may be applied in patient populations with intact sensation such as stroke and amyotrophic lateral sclerosis to restore an effective cough.
Melatonin Supplementation Lowers Oxidative Stress and Regulates Adipokines in Obese Patients on a Calorie‐Restricted Diet
Authors: Szewczyk-Golec Karolina , Rajewski Paweł , Gackowski Marcin , Mila-Kierzenkowska Celestyna , Wesołowski Roland , Sutkowy Paweł , Pawłowska Marta , Woźniak Alina
Published: September 2017

Obesity is one of the major global health problems. Melatonin deficiency has been demonstrated to correlate with obesity. The aim of the study was to estimate the effect of melatonin on oxidative stress and adipokine levels in obese patients on a calorie‐restricted diet. Thirty obese patients were supplemented with a daily dose of 10 mg of melatonin (n = 15) or placebo (n = 15) for 30 days with a calorie‐restricted diet. Serum levels of melatonin, 4‐hydroxynonenal (HNE), adiponectin, omentin‐1, leptin, and resistin, as well as erythrocytic malondialdehyde (MDA) concentration and Zn/Cu‐superoxide dismutase, catalase, and glutathione peroxidase (GPx) activities, were measured at baseline and after supplementation. Significant body weight reduction was observed only in the melatonin group. After melatonin supplementation, the adiponectin and omentin‐1 levels and GPx activities statistically increased, whereas the MDA concentrations were reduced. In the placebo group, a significant rise in the HNE and a drop in the melatonin concentrations were found. The results show evidence of increased oxidative stress accompanying calorie restriction. Melatonin supplementation facilitated body weight reduction, improved the antioxidant defense, and regulated adipokine secretion. The findings strongly suggest that melatonin should be considered in obesity management. This trial is registered with CTRI/2017/07/009093.
Are You What You Eat or What Your Mother Ate or Both?
Authors: Freedland Stephen J.
Published: September 2017

A high-fat high-sugar (HF-HS) diet promotes cancer development and progression. However, does the timing of diet matter? This is an important question with profound public health relevance. By exposing mice to a HF-HS diet either through feeding to a pregnant mother or nursing mother or after weaning and then chemically inducing breast cancer, the authors found the most crucial time for breast cancer risk was after weaning, while a HF-HS in utero diet actually slowed tumor development. Understanding early-life events provides valuable insight for later life events and proves it is never too early to start preventing disease. Cancer Prev Res; 10(10); 551–2. ©2017 AACR.

See related article by Lambertz, p. 553–62.
Early Electroencephalography Suppression and Postnatal Morbidities Correlate with Cerebral Volume at Term-Equivalent Age in Very Preterm Infants
Authors: Wikström Sverre , Hövel Holger , Hansen Pupp Ingrid , Fellman Vineta , Hüppi Petra S. , Ley David , Hellström-Westas Lena
Published: September 2017

Background: Early brain activity is associated with long-term outcome. Establishing a relation also with postnatal brain growth may increase our understanding of early life influences on preterm brain development. Objectives: The aim of this study was to investigate whether early electroencephalography (EEG) activity in infants born very preterm is associated with brain volumes at term, and whether postnatal morbidity affects this association. Methods: Very preterm infants (n = 38) with a median gestational age (GA) of 25.6 weeks had early recordings of single-channel EEG. The percentage of suppressed EEG, i.e., interburst intervals (IBI%) between 24 and 72 h of age, was analyzed in relation to brain volumes on magnetic resonance imaging performed at term-equivalent age, taking into account neonatal morbidities. Results: Early electrocortical depression and a higher IBI% were associated with increased cerebrospinal fluid volume (CSFV) and lower total brain volume relative to intracranial volume, also after adjustment for GA, postnatal morbidities, morphine administration, and postnatal head growth. Overall, an increase in IBI% to 1 SD from the mean corresponded with an increase in CSFV to +0.7 SD and a decrease in brain volume to -0.7 SD. The presence of 2 or more postnatal morbidities were associated with around 10% lower brain volumes. Conclusions: More suppressed early EEG activity of very preterm infants is associated with lower brain volume and increased CSFV at term age, also when adjusting for postnatal morbidities. The findings indicate the importance of pre- and early postpartal determinants of postnatal brain growth, possibly also including activity-dependent mechanisms for brain growth.
Successful Management of a Positive Air Leak Test during Laparoscopic Colorectal Surgery
Authors: Sasaki Kazuhito , Ishihara Soichiro , Nozawa Hiroaki , Kawai Kazushige , Hata Keisuke , Kiyomatsu Tomomichi , Tanaka Toshiaki , Nishikawa Takeshi , Otani Kensuke , Yasuda Koji , Murono Koji , Watanabe Toshiaki
Published: September 2017

Background/Aims: Anastomotic leakage remains the most serious complications of colorectal surgery. To prevent colorectal anastomotic leakage (CAL), an air leak test (ALT) with intraoperative colonoscopy (IOCS) is performed to detect mechanically insufficient colorectal anastomoses. The approaches to an intraoperative anastomotic air leak (IOAL) have not been fully investigated. This study aimed to clarify the safe management of an IOAL in laparoscopic colorectal surgery. Methods: One hundred forty-eight consecutive patients who underwent laparoscopic resection with double-stapling technique (DST) anastomosis for left-sided colorectal cancer between April 2015 and June 2016 were included and retrospectively reviewed. Results: Intraoperative anastomotic ALT yielded positive results in 7 patients. In all 7 patients, reanastomoses were performed, and diverting stomas were constructed to protect the anastomosis in 2 patients whose reanastomosis sites were close to the anus. Three of the revised DST anastomoses showed air leakage on the repeat ALT; these sites underwent suturing repair and were confirmed to be airtight. None of the patients with a positive intraoperative ALT had postoperative CAL. The overall CAL rate was 1.4%. Conclusions: Combination management using DST revision, direct suturing repair, and a diverting stoma is recommended for intraoperative repair of anastomotic defects detected by IOCS.