Novel molecular insights into the function and the antioxidative stress response of a Peroxiredoxin Q protein from Cyanobacteria

Publication date: Available online 31 January 2017
Source:Free Radical Biology and Medicine
Author(s): Vijay Tailor, Anand Ballal
The PeroxiredoxinQ (PrxQ) proteins are thiol-based peroxidases that are important for maintaining redox homeostasis in several organisms. Activity of PrxQs is mediated by two cysteines, peroxidatic (Cp) and resolving (Cr), in association with a reducing partner. A PrxQ, Alr3183, from the cyanobacterium, Anabaena PCC 7120, was characterized in this study. Alr3183, which required thioredoxin A (TrxA) for peroxidase activity, was an intramolecular disulfide bond-containing monomeric protein. However, Alr3183 lacking Cp (Alr3183C46S) or Cr (Alr3183C51S) formed intermolecular disulphide linkages and was dimeric. Alr3183C46S was completely inactive, while Alr3183C51S required higher concentration of TrxA for peroxidase activity. Surface plasmon resonance analysis showed that unlike Alr3183 or Alr3183C46S, Alr3183C51S bound rather poorly to TrxA. Also, compared to the oxidized protein, the DTT-treated (reduced) Alr3183 displayed decreased interaction with TrxA. In vivo, Alr3183 was found to be induced in response to γ-radiation. On exposure to H2O2, Anabaena strain over-expressing Alr3183 showed reduced formation of ROS, intact photosynthetic pigments and consequently better survival than the wild-type, whereas overproduction of Alr3183C46S did not provide any protection. Significantly, this study (1) reveals the importance of Cr for interaction with thioredoxins and (2) demonstrates that over-expression of PrxQs can protect cyanobacteria from oxidative stresses.

Graphical abstract

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Combined p16 and p53 expression in cervical cancer of unknown primary and other prognostic parameters

Abstract

Background and purpose

Cervical cancer of unknown primary (CUP) represents an uncommon and heterogeneous subentity of head and neck cancer. However, both optimal diagnostics and therapy remain unclear. An improved understanding of the underlying pathology is essential to enable future tailored therapies and optimized outcomes.

Materials and methods

We retrospectively analyzed 53 patients with head and neck CUP and 48 available cervical lymph node specimens. All patients have received radiotherapy between 2007 and 2015. Preradiotherapy involved lymph node specimens were analyzed for p16 and p53 immunoreactivity. The prognostic relevance of the combined p16 and p53 status and other clinical parameters were examined by univariate and multivariate analyses.

Results

Median patient age was 61.5 years and median irradiation dose to the involved nodal levels was 66 Gy. Of the 48 evaluated specimens, 13 (27%) were p16-positive and 31 (64.6%) p53-positive. After a median follow up of 32.9 months, patients with p16-negative and simultaneously p53-positive tumors showed a significantly inferior tumor-specific survival (TSS) compared to those with either p16+/p53−, p16+/p53+, or p16−/p53− (univariate: p = 0.055, multivariate: p = 0.038). Other factors with an adverse impact on TSS in the univariate analysis were smoking history (p = 0.032) and nodal stage (p = 0.038).

Conclusions

The combined p16- and p53-expression status in cervical metastases of CUP may represent a simple method for risk stratification. Further validation of these biomarkers in large prospective trials is essential to design rational trials for CUP treatment optimization.

http://ift.tt/2kRA9Jy

Functional evaluation of the restored mucosa after nasal reconstruction with a forehead-galea flap

Abstract

Background

Recovery of the internal mucosal lining is the most problematic step in nasal reconstruction. Restoration of both aesthetic and functional components should be the goal to be pursued. For this purpose, we performed a study for functional evaluation of the restored mucosa after nasal reconstruction.

Methods

From April 2009 to May 2016, 10 patients in whom the galea was used to reconstruct the nasal lining were selected from our casuistic of nasal reconstruction. In order to visualize the nasal and nasopharyngeal cavity, an antero-posterior rhinoendoscopy was performed in all patients. Additionally, patients were asked to complete a visual analogue scale (VAS) evaluation regarding nasal obstruction. Active anterior rhinomanometry analysis, olfactometry analysis and a cytologic examination were also conducted.

Results

Near-normal results in nasal obstruction evaluation were reported subjectively by patients. Near-normal inspiratory values were obtained using rhinomanometry. Average values of TDI (threshold, discrimination and identification), a comprehensive olfactometric parameter, were essentially normal. Cytological sampling examination did not reveal any substantial abnormal variation.

Conclusions

Based on our morphological and functional results, we can assert that the forehead flap in association with galea for lining is a safe and practical surgical technique in total nasal reconstruction.

Level of Evidence: Level V, prognostic study

http://ift.tt/2klLJ2i

Molecular cloning, characterization and expression analysis of Tim-3 and Galectin-9 in the woodchuck model

Publication date: March 2017
Source:Molecular Immunology, Volume 83
Author(s): Yanan Liu, Junzhong Wang, Lu Wang, Baoju Wang, Shangqing Yang, Qin Wang, Jinzhuo Luo, Xuemei Feng, Xuecheng Yang, Yinping Lu, Michael Roggendorf, Mengji Lu, Dongliang Yang, Jia Liu
In recent years, a critical role for T cell immunoglobulin mucin domain 3 (Tim-3) and its ligand Galectin-9 (Gal-9) has emerged in infectious disease, autoimmunity and cancer. Manipulating this immune checkpoint may have immunotherapeutic potential and could represent an alternative approach for improving immune responses to viral infections and cancer. The woodchuck (Marmot monax) infected by woodchuck hepatitis virus (WHV) represents an informative animal model to study HBV infection and HCC. In the current study, the cDNA sequences of woodchuck Tim-3 and Gal-9 were cloned, sequenced and characterized. The extracellular domain of Tim-3 cDNA sequence consisted of 576bp coding sequence (CDS) that encoded 192 amino acids. The 1076bp full-length Gal-9 cDNA sequence consisted of 1059bp coding sequence (CDS) that encoded 352 amino acids with a molecular weight of 39.7kDa. The phylogenetic tree analysis revealed that the woodchuck Tim-3 and Gal-9 had the closest genetic relationship with Ictidomys tridecemlineatus. The result of quantification PCR analysis showed that ubiquitous expression of Gal-9 but not Tim-3 in different tissues of naive woodchucks. Elevated liver Gal-9 expression was observed in woodchucks with chronic WHV infection. Moreover, a polyclonal antibody against the extracellular domain of woodchuck Tim-3 were generated and identified by flow cytometry. Our results serve as a foundation for further insight into the role of Tim-3/Galectin-9 signaling pathway in viral hepatitis and HCC in the woodchuck model.



http://ift.tt/2jzLa18

DnaJ (hsp40) of Streptococcus pneumoniae is involved in bacterial virulence and elicits a strong natural immune reaction via PI3K/JNK

Publication date: March 2017
Source:Molecular Immunology, Volume 83
Author(s): Jin Cui, Chenyu Ma, Guo Ye, Yong Shi, Wenchun Xu, Liang Zhong, Jian Wang, Yibing Yin, Xuemei Zhang, Hong Wang
As a heat shock protein, DnaJ plays an important role in the pathogenesis of pneumococcal infection. However, how the virulence factor-DnaJ elicits host natural immunity still remains unclear. In this study, we investigated the effects of dnaJ deficiency in Streptococcus pneumoniae (S. pneumoniae) on bacterial virulence, and further explored the related molecular mechanisms in vivo and in vitro. By generating dnaJ deficient mutant (ΔdnaJ), the virulence and colonization were detected in murine pneumonia and sepsis models in vivo. Compared with wild-type parent strain, the abilities of rapid colonization and induction of inflammatory responses of ΔdnaJ in mouse lungs were significantly impaired. Simultaneously, recombinant DnaJ purified from E. coli expression system (rDnaJ) induced macrophage strain RAW264.7 to secrete IL-6 by activation of PI3K and JNK signal pathways, which were confirmed by the specific signaling inhibitors. In conclusion, DnaJ, a novel virulence protein, was essential for the virulence and colonization of S. pneumoniae and induced pro-inflammatory cytokine production in macrophages through PI3K/JNK.



http://ift.tt/2knK6Sv

Immunoproteasome subunit deficiency has no influence on the canonical pathway of NF-κB activation

Publication date: March 2017
Source:Molecular Immunology, Volume 83
Author(s): Annegret Bitzer, Michael Basler, Daniel Krappmann, Marcus Groettrup
Activation of the pro-inflammatory transcription factor NF-κB requires signal-induced proteasomal degradation of the inhibitor of NF-κB (IκB) in order to allow nuclear translocation. Most cell types are capable of expressing two types of 20S proteasome core particles, the constitutive proteasome and immunoproteasome. Inducible under inflammatory conditions, the immunoproteasome is mainly characterized through an altered cleavage specificity compared to the constitutive proteasome. However, the question whether immunoproteasome subunits affect NF-κB signal transduction differently from constitutive subunits is still up for debate. To study the effect of immunoproteasomes on LPS- or TNF-α-induced NF-κB activation, we used IFN-γ stimulated peritoneal macrophages and mouse embryonic fibroblasts derived from mice deficient for the immunoproteasome subunits low molecular mass polypeptide (LMP) 2, or LMP7 and multicatalytic endopeptidase complex-like 1 (MECL-1). Along the canonical signaling pathway of NF-κB activation no differences in the extent and kinetic of IκB degradation were observed. Neither the nuclear translocation and DNA binding of NF-κB nor the production of the NF-κB dependent cytokines TNF-α, IL-6, and IL-10 differed between immunoproteasome deficient and proficient cells. Hence, we conclude that immunoproteasome subunits have no specialized function for canonical NF-κB activation.

Graphical abstract

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Relationship between the change of language symptoms and the change of regional cerebral blood flow in the recovery process of two children with acquired aphasia

Publication date: Available online 31 January 2017
Source:Brain and Development
Author(s): Junko Kozuka, Akira Uno, Hiroshi Matsuda, Yoshiya Toyoshima, Shin-ichiro Hamano
ObjectiveThis study aimed to investigate the relationship between the change of language symptoms and the change of regional cerebral blood flow (rCBF) in the recovery process of two children with acquired aphasia caused by infarctions from Moyamoya disease with an onset age of 8years.MethodsWe compared the results for the Standard Language Test of Aphasia (SLTA) with rCBF changes in 7 language regions in the left hemisphere and their homologous regions in the right hemisphere at 4 time points from 3weeks for up to 5years after the onset of aphasia, while controlling for the effect of age.ResultsIn both cases, strong correlations were seen within a hemisphere between adjacent regions or regions that are connected by neuronal fibers, and between some language regions in the left hemisphere and their homologous regions in the right hemisphere. Conversely, there were differences between the two cases in the time course of rCBF changes during their recovery process.ConclusionConsistent with previous studies, the current study suggested that both hemispheres were involved in the long-term recovery of language symptoms in children with acquired aphasia. We suggest that the differences between both cases during their recovery process might be influenced by the brain states before aphasia, by which hemisphere was affected, and by the timing of the surgical revascularization procedure. However, the changes were observed in the data obtained for rCBF with strong correlations with the changes in language performance, so it is possible that rCBF could be used as a biomarker for language symptom changes.



http://ift.tt/2jzDwDG

Visualizing Changes in Cdkn1c Expression Links Early-Life Adversity to Imprint Mis-regulation in Adults

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Mathew Van de Pette, Allifia Abbas, Amelie Feytout, Gráinne McNamara, Ludovica Bruno, Wilson K. To, Andrew Dimond, Alessandro Sardini, Zoe Webster, James McGinty, Eleanor J. Paul, Mark A. Ungless, Paul M.W. French, Dominic J. Withers, Anthony Uren, Anne C. Ferguson-Smith, Matthias Merkenschlager, Rosalind M. John, Amanda G. Fisher
Imprinted genes are regulated according to parental origin and can influence embryonic growth and metabolism and confer disease susceptibility. Here, we designed sensitive allele-specific reporters to non-invasively monitor imprinted Cdkn1c expression in mice and showed that expression was modulated by environmental factors encountered in utero. Acute exposure to chromatin-modifying drugs resulted in de-repression of paternally inherited (silent) Cdkn1c alleles in embryos that was temporary and resolved after birth. In contrast, deprivation of maternal dietary protein in utero provoked permanent de-repression of imprinted Cdkn1c expression that was sustained into adulthood and occurred through a folate-dependent mechanism of DNA methylation loss. Given the function of imprinted genes in regulating behavior and metabolic processes in adults, these results establish imprinting deregulation as a credible mechanism linking early-life adversity to later-life outcomes. Furthermore, Cdkn1c-luciferase mice offer non-invasive tools to identify factors that disrupt epigenetic processes and strategies to limit their long-term impact.

Graphical abstract

Teaser

Van de Pette et al. use sensitive allele-specific reporters to longitudinally image imprinted Cdkn1c expression in mice and show that expression is modulated by environmental factors encountered in utero. These results establish imprinting deregulation as a mechanism linking early-life adversity to later-life outcomes and provide tools to detect imprinting changes in vivo.


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Abnormal Development of the Earliest Cortical Circuits in a Mouse Model of Autism Spectrum Disorder

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Daniel A. Nagode, Xiangying Meng, Daniel E. Winkowski, Ed Smith, Hamza Khan-Tareen, Vishnupriya Kareddy, Joseph P.Y. Kao, Patrick O. Kanold
Autism spectrum disorder (ASD) involves deficits in speech and sound processing. Cortical circuit changes during early development likely contribute to such deficits. Subplate neurons (SPNs) form the earliest cortical microcircuits and are required for normal development of thalamocortical and intracortical circuits. Prenatal valproic acid (VPA) increases ASD risk, especially when present during a critical time window coinciding with SPN genesis. Using optical circuit mapping in mouse auditory cortex, we find that VPA exposure on E12 altered the functional excitatory and inhibitory connectivity of SPNs. Circuit changes manifested as "patches" of mostly increased connection probability or strength in the first postnatal week and as general hyper-connectivity after P10, shortly after ear opening. These results suggest that prenatal VPA exposure severely affects the developmental trajectory of cortical circuits and that sensory-driven activity may exacerbate earlier, subtle connectivity deficits. Our findings identify the subplate as a possible common pathophysiological substrate of deficits in ASD.

Graphical abstract

Teaser

It has been hypothesized that dysfunction of subplate neurons is an early event in autism pathology, but never directly tested. Nagode et al. demonstrate spatially restricted increases in excitatory and inhibitory connectivity to subplate in the VPA autism model. These results provide direct evidence of subplate dysfunction in autism.


http://ift.tt/2kN2YaD

Autonomous CaMKII Activity as a Drug Target for Histological and Functional Neuroprotection after Resuscitation from Cardiac Arrest

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Guiying Deng, James E. Orfila, Robert M. Dietz, Myriam Moreno-Garcia, Krista M. Rodgers, Steve J. Coultrap, Nidia Quillinan, Richard J. Traystman, K. Ulrich Bayer, Paco S. Herson
The Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) is a major mediator of physiological glutamate signaling, but its role in pathological glutamate signaling (excitotoxicity) remains less clear, with indications for both neuro-toxic and neuro-protective functions. Here, the role of CaMKII in ischemic injury is assessed utilizing our mouse model of cardiac arrest and cardiopulmonary resuscitation (CA/CPR). CaMKII inhibition (with tatCN21 or tatCN19o) at clinically relevant time points (30 min after resuscitation) greatly reduces neuronal injury. Importantly, CaMKII inhibition also works in combination with mild hypothermia, the current standard of care. The relevant drug target is specifically Ca²⁺-independent "autonomous" CaMKII activity generated by T286 autophosphorylation, as indicated by substantial reduction in injury in autonomy-incompetent T286A mutant mice. In addition to reducing cell death, tatCN19o also protects the surviving neurons from functional plasticity impairments and prevents behavioral learning deficits, even at extremely low doses (0.01 mg/kg), further highlighting the clinical potential of our findings.

Graphical abstract

Teaser

Deng et al. find that CaMKII and its phospho-T286-induced autonomous activity are a promising therapeutic drug target for global cerebral ischemia (induced by cardiac arrest followed by CPR in a mouse model). Pharmacological inhibition or T286A mutation leads to neuroprotection and improves synaptic and functional recovery following cardiac arrest.


http://ift.tt/2kMIOxq

Altered Tau Isoform Ratio Caused by Loss of FUS and SFPQ Function Leads to FTLD-like Phenotypes

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Shinsuke Ishigaki, Yusuke Fujioka, Yohei Okada, Yuichi Riku, Tsuyoshi Udagawa, Daiyu Honda, Satoshi Yokoi, Kuniyuki Endo, Kensuke Ikenaka, Shinnosuke Takagi, Yohei Iguchi, Naruhiko Sahara, Akihiko Takashima, Hideyuki Okano, Mari Yoshida, Hitoshi Warita, Masashi Aoki, Hirohisa Watanabe, Haruo Okado, Masahisa Katsuno, Gen Sobue
Fused in sarcoma (FUS) and splicing factor, proline- and glutamine-rich (SFPQ) are RNA binding proteins that regulate RNA metabolism. We found that alternative splicing of the Mapt gene at exon 10, which generates 4-repeat tau (4R-T) and 3-repeat tau (3R-T), is regulated by interactions between FUS and SFPQ in the nuclei of neurons. Hippocampus-specific FUS- or SFPQ-knockdown mice exhibit frontotemporal lobar degeneration (FTLD)-like behaviors, reduced adult neurogenesis, accumulation of phosphorylated tau, and hippocampal atrophy with neuronal loss through an increased 4R-T/3R-T ratio. Normalization of this increased ratio by 4R-T-specific silencing results in recovery of the normal phenotype. These findings suggest a biological link among FUS/SFPQ, tau isoform alteration, and phenotypic expression, which may function in the early pathomechanism of FTLD.

Graphical abstract

Teaser

Ishigaki et al. investigate the functions of Fused in sarcoma (FUS) and its binding partner proline- and glutamine-rich (SFPQ) in regulation of Mapt splicing. The authors show how loss of interaction between FUS and SFPQ causes altered expression ratio of tau isoforms and leads to a neurodegenerative phenotype similar to FTLD. Normalization of this ratio can reduce phenotypic abnormalities in the mouse model of disease.


http://ift.tt/2kMIy1z

Auxilin Underlies Progressive Locomotor Deficits and Dopaminergic Neuron Loss in a Drosophila Model of Parkinson’s Disease

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Li Song, Yijing He, Jiayao Ou, Yongbo Zhao, Ruoyu Li, Jingjing Cheng, Chin-Hsien Lin, Margaret S. Ho
Parkinson's disease (PD) is a common neurodegenerative disorder that exhibits motor and non-motor symptoms, as well as pathological hallmarks, including dopaminergic (DA) neuron death and formation of α-synuclein (α-Syn) Lewy bodies. Cyclin-G-associated kinase (GAK), a PD susceptibility gene identified through genome-wide association studies (GWAS), is a ubiquitous serine/threonine kinase involved in clathrin uncoating, though its PD-related function remains elusive. Here, we implicate the Drosophila GAK homolog, auxilin (aux), in a broad spectrum of parkinsonian-like symptoms. Downregulating aux expression leads to progressive loss of climbing ability, decreased lifespan, and age-dependent DA neuron death similar to α-Syn overexpression. Reduced aux expression further enhances and accelerates α-Syn-mediated DA neuron loss. Flies with reduced aux expression are more sensitive to the toxin paraquat, suggesting that genetic and environmental factors intertwine. Taken together, these findings decipher a pivotal role for GAK/aux and suggest mechanisms underlying PD.

Graphical abstract

Teaser

Parkinson's disease (PD) is a neurodegenerative disorder that exhibits motor symptoms and neuron loss. Song et al. implicate Drosophila cyclin-G-associated kinase (GAK) in fly phenotypes reminiscent of PD.


http://ift.tt/2kMJUJP

Gender Differences in Global but Not Targeted Demethylation in iPSC Reprogramming

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Inês Milagre, Thomas M. Stubbs, Michelle R. King, Julia Spindel, Fátima Santos, Felix Krueger, Martin Bachman, Anne Segonds-Pichon, Shankar Balasubramanian, Simon R. Andrews, Wendy Dean, Wolf Reik
Global DNA demethylation is an integral part of reprogramming processes in vivo and in vitro, but whether it occurs in the derivation of induced pluripotent stem cells (iPSCs) is not known. Here, we show that iPSC reprogramming involves both global and targeted demethylation, which are separable mechanistically and by their biological outcomes. Cells at intermediate-late stages of reprogramming undergo transient genome-wide demethylation, which is more pronounced in female cells. Global demethylation requires activation-induced cytidine deaminase (AID)-mediated downregulation of UHRF1 protein, and abolishing demethylation leaves thousands of hypermethylated regions in the iPSC genome. Independently of AID and global demethylation, regulatory regions, particularly ESC enhancers and super-enhancers, are specifically targeted for hypomethylation in association with transcription of the pluripotency network. Our results show that global and targeted DNA demethylation are conserved and distinct reprogramming processes, presumably because of their respective roles in epigenetic memory erasure and in the establishment of cell identity.

Graphical abstract

Teaser

Milagre et al. find that two modes of DNA demethylation occur during primary iPSC reprogramming. Global DNA demethylation, more pronounced in female cells, is regulated by AID through UHRF1 and occurs transiently at intermediate-late stages of reprogramming. Targeted DNA demethylation, by contrast, is important in establishing hypomethylation at enhancers of pluripotency genes and occurs similarly in female and male cells.


http://ift.tt/2kMRhRi

Serotonin Signaling through Prefrontal Cortex 5-HT1A Receptors during Adolescence Can Determine Baseline Mood-Related Behaviors

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Alvaro L. Garcia-Garcia, Qingyuan Meng, Sarah Canetta, Alain M. Gardier, Bruno P. Guiard, Christoph Kellendonk, Alex Dranovsky, E. David Leonardo
Lifelong homeostatic setpoints for mood-related behaviors emerge during adolescence. Serotonin (5-HT) plays an important role in refining the formation of brain circuits during sensitive developmental periods. In rodents, the role of 5-HT1A receptors in general and autoreceptors in particular has been characterized in anxiety. However, less is known about the role of 5-HT1A receptors in depression-related behavior. Here, we show that whole-life suppression of heteroreceptor expression results in a broad depression-like behavioral phenotype accompanied by physiological and cellular changes within medial prefrontal cortex-dorsal raphe proper (mPFC-DRN) circuitry. These changes include increased basal 5-HT in a mPFC that is hyporesponsive to stress and decreased basal 5-HT levels and firing rates in a DRN hyperactivated by the same stressor. Remarkably, loss of heteroreceptors in the PFC at adolescence is sufficient to recapitulate this depression-like behavioral syndrome. Our results suggest that targeting mPFC 5-HT1A heteroreceptors during adolescence in humans may have lifelong ramifications for depression and its treatment.

Graphical abstract

Teaser

Garcia-Garcia et al. use transgenic and viral approaches to demonstrate that signaling through 5-HT1A heteroreceptors in the medial prefrontal cortex during adolescence is critical in establishing baseline mood setpoints.


http://ift.tt/2kMQIan

Neuregulin 3 Mediates Cortical Plate Invasion and Laminar Allocation of GABAergic Interneurons

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Giorgia Bartolini, Juan Antonio Sánchez-Alcañiz, Catarina Osório, Manuel Valiente, Cristina García-Frigola, Oscar Marín
Neural circuits in the cerebral cortex consist of excitatory pyramidal cells and inhibitory interneurons. These two main classes of cortical neurons follow largely different genetic programs, yet they assemble into highly specialized circuits during development following a very precise choreography. Previous studies have shown that signals produced by pyramidal cells influence the migration of cortical interneurons, but the molecular nature of these factors has remained elusive. Here, we identified Neuregulin 3 (Nrg3) as a chemoattractive factor expressed by developing pyramidal cells that guides the allocation of cortical interneurons in the developing cortical plate. Gain- and loss-of-function approaches reveal that Nrg3 modulates the migration of interneurons into the cortical plate in a process that is dependent on the tyrosine kinase receptor ErbB4. Perturbation of Nrg3 signaling in conditional mutants leads to abnormal lamination of cortical interneurons. Nrg3 is therefore a critical mediator in the assembly of cortical inhibitory circuits.

Graphical abstract

Teaser

The integration of interneurons in the developing cerebral cortex depends on signals generated by pyramidal cells. In this study, Bartolini et al. identify Neuregulin 3 as a developmentally regulated factor that controls the migration of interneurons into the cortical plate and influences their final laminar distribution.


http://ift.tt/2kMIL4W

miR-182 Regulates Slit2-Mediated Axon Guidance by Modulating the Local Translation of a Specific mRNA

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Anaïs Bellon, Archana Iyer, Simone Bridi, Flora C.Y. Lee, Cesaré Ovando-Vázquez, Eloina Corradi, Sara Longhi, Michela Roccuzzo, Stephanie Strohbuecker, Sindhu Naik, Peter Sarkies, Eric Miska, Cei Abreu-Goodger, Christine E. Holt, Marie-Laure Baudet
During brain wiring, cue-induced axon behaviors such as directional steering and branching are aided by localized mRNA translation. Different guidance cues elicit translation of subsets of mRNAs that differentially regulate the cytoskeleton, yet little is understood about how specific mRNAs are selected for translation. MicroRNAs (miRNAs) are critical translational regulators that act through a sequence-specific mechanism. Here, we investigate the local role of miRNAs in mRNA-specific translation during pathfinding of Xenopus laevis retinal ganglion cell (RGC) axons. Among a rich repertoire of axonal miRNAs, miR-182 is identified as the most abundant. Loss of miR-182 causes RGC axon targeting defects in vivo and impairs Slit2-induced growth cone (GC) repulsion. We find that miR-182 targets cofilin-1 mRNA, silencing its translation, and Slit2 rapidly relieves the repression without causing miR-182 degradation. Our data support a model whereby miR-182 reversibly gates the selection of transcripts for fast translation depending on the extrinsic cue.

Graphical abstract

Teaser

Bellon et al. examine how specific mRNAs are selected for cue-induced local protein synthesis during axon guidance and find that miR-182 reversibly regulates the selective translation of a specific transcript to facilitate fast growth cone steering and axon guidance.


http://ift.tt/2kMP1JJ

Mechanism of β-actin mRNA Recognition by ZBP1

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Giuseppe Nicastro, Adela M. Candel, Michael Uhl, Alain Oregioni, David Hollingworth, Rolf Backofen, Stephen R. Martin, Andres Ramos
Zipcode binding protein 1 (ZBP1) is an oncofetal RNA-binding protein that mediates the transport and local translation of β-actin mRNA by the KH3-KH4 di-domain, which is essential for neuronal development. The high-resolution structures of KH3-KH4 with their respective target sequences show that KH4 recognizes a non-canonical GGA sequence via an enlarged and dynamic hydrophobic groove, whereas KH3 binding to a core CA sequence occurs with low specificity. A data-informed kinetic simulation of the two-step binding reaction reveals that the overall reaction is driven by the second binding event and that the moderate affinities of the individual interactions favor RNA looping. Furthermore, the concentration of ZBP1, but not of the target RNA, modulates the interaction, which explains the functional significance of enhanced ZBP1 expression during embryonic development.

Graphical abstract

Teaser

The interaction of β-actin mRNA with the RNA-binding protein ZBP1 regulates mRNA transport and translation and is necessary for neuronal development. Using NMR and biophysics, Nicastro et al. have shown that the interaction is driven by RNA looping and regulated by the protein, rather than the RNA, concentration.


http://ift.tt/2kMSU1x

Visualization of Chromatin Decompaction and Break Site Extrusion as Predicted by Statistical Polymer Modeling of Single-Locus Trajectories

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Assaf Amitai, Andrew Seeber, Susan M. Gasser, David Holcman
Chromatin moves with subdiffusive and spatially constrained dynamics within the cell nucleus. Here, we use single-locus tracking by time-lapse fluorescence microscopy to uncover information regarding the forces that influence chromatin movement following the induction of a persistent DNA double-strand break (DSB). Using improved time-lapse imaging regimens, we monitor trajectories of tagged DNA loci at a high temporal resolution, which allows us to extract biophysical parameters through robust statistical analysis. Polymer modeling based on these parameters predicts chromatin domain expansion near a DSB and damage extrusion from the domain. Both phenomena are confirmed by live imaging in budding yeast. Calculation of the anomalous exponent of locus movement allows us to differentiate forces imposed on the nucleus through the actin cytoskeleton from those that arise from INO80 remodeler-dependent changes in nucleosome organization. Our analytical approach can be applied to high-density single-locus trajectories obtained in any cell type.

Graphical abstract

Teaser

Amitai et al. present a robust analytical workflow for the analysis of single-particle trajectories. The extracted biophysical parameters allow accurate modeling of chromatin dynamics. The authors predict and show that, at double-strand breaks, chromatin decompacts and induces break relocation, while nuclear oscillation arises from cytoskeleton forces.


http://ift.tt/2kMP1tj

Mesenchymal Stem Cell-Induced DDR2 Mediates Stromal-Breast Cancer Interactions and Metastasis Growth

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Maria E. Gonzalez, Emily E. Martin, Talha Anwar, Caroline Arellano-Garcia, Natasha Medhora, Arjun Lama, Yu-Chih Chen, Kevin S. Tanager, Euisik Yoon, Kelley M. Kidwell, Chunxi Ge, Renny T. Franceschi, Celina G. Kleer
Increased collagen deposition by breast cancer (BC)-associated mesenchymal stem/multipotent stromal cells (MSC) promotes metastasis, but the mechanisms are unknown. Here, we report that the collagen receptor discoidin domain receptor 2 (DDR2) is essential for stromal-BC communication. In human BC metastasis, DDR2 is concordantly upregulated in metastatic cancer and multipotent mesenchymal stromal cells. In MSCs isolated from human BC metastasis, DDR2 maintains a fibroblastic phenotype with collagen deposition and induces pathological activation of DDR2 signaling in BC cells. Loss of DDR2 in MSCs impairs their ability to promote DDR2 phosphorylation in BC cells, as well as BC cell alignment, migration, and metastasis. Female ddr2-deficient mice homozygous for the slie mutation show inefficient spontaneous BC metastasis. These results point to a role for mesenchymal stem cell DDR2 in metastasis and suggest a therapeutic approach for metastatic BC.

Graphical abstract

Teaser

By isolating human mesenchymal stem cells from breast cancer metastasis, Gonzalez et al. identify a pathway initiated by stromal DDR2, a unique receptor tyrosine kinase activated by fibrillar collagen, that mediates DDR2 activation in breast cancer cells and induces metastasis.


http://ift.tt/2kMQ9NQ

Regulation of Serine-Threonine Kinase Akt Activation by NAD+-Dependent Deacetylase SIRT7

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Jia Yu, Bo Qin, Fengying Wu, Sisi Qin, Somaira Nowsheen, Shan Shan, Jacqueline Zayas, Huadong Pei, Zhenkun Lou, Liewei Wang
The Akt pathway is a central regulator that promotes cell survival in response to extracellular signals. Depletion of SIRT7, an NAD⁺-dependent deacetylase that is the least-studied sirtuin, is known to significantly increase Akt activity in mice through unknown mechanisms. In this study, we demonstrate that SIRT7 depletion in breast cancer cells results in Akt hyper-phosphorylation and increases cell survival following genotoxic stress. Mechanistically, SIRT7 specifically interacts with and deacetylates FKBP51 at residue lysines 28 and 155 (K28 and K155), resulting in enhanced interactions among FKBP51, Akt, and PHLPP, as well as Akt dephosphorylation. Mutating both lysines to arginines abolishes the effect of SIRT7 on Akt activity through FKBP51 deacetylation. Finally, energy stress strengthens SIRT7-mediated effects on Akt dephosphorylation through FKBP51 and thus sensitizes cancer cells to cytotoxic agents. These results reveal a direct role of SIRT7 in Akt regulation and raise the possibility of using the glucose analog 2-deoxy-D-glucose (2DG) as a chemo-sensitizing agent.

Graphical abstract

Teaser

Yu et al. reveal a direct role of SIRT7 in regulating Akt activation and affecting cancer cell responses to chemotherapies under energy stress.


http://ift.tt/2kMIPkY

The Nuclear Receptor LXR Limits Bacterial Infection of Host Macrophages through a Mechanism that Impacts Cellular NAD Metabolism

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Jonathan Matalonga, Estibaliz Glaria, Mariana Bresque, Carlos Escande, José María Carbó, Kerstin Kiefer, Ruben Vicente, Theresa E. León, Susana Beceiro, Mónica Pascual-García, Joan Serret, Lucía Sanjurjo, Samantha Morón-Ros, Antoni Riera, Sonia Paytubi, Antonio Juarez, Fernando Sotillo, Lennart Lindbom, Carme Caelles, Maria-Rosa Sarrias, Jaime Sancho, Antonio Castrillo, Eduardo N. Chini, Annabel F. Valledor
Macrophages exert potent effector functions against invading microorganisms but constitute, paradoxically, a preferential niche for many bacterial strains to replicate. Using a model of infection by Salmonella Typhimurium, we have identified a molecular mechanism regulated by the nuclear receptor LXR that limits infection of host macrophages through transcriptional activation of the multifunctional enzyme CD38. LXR agonists reduced the intracellular levels of NAD⁺ in a CD38-dependent manner, counteracting pathogen-induced changes in macrophage morphology and the distribution of the F-actin cytoskeleton and reducing the capability of non-opsonized Salmonella to infect macrophages. Remarkably, pharmacological treatment with an LXR agonist ameliorated clinical signs associated with Salmonella infection in vivo, and these effects were dependent on CD38 expression in bone-marrow-derived cells. Altogether, this work reveals an unappreciated role for CD38 in bacterial-host cell interaction that can be pharmacologically exploited by activation of the LXR pathway.

Graphical abstract

Teaser

Macrophages constitute a preferential niche for intracellular bacteria to replicate. Matalonga et al. identified an antibacterial circuit mediated by pharmacological targeting of the liver X receptor (LXR) pathway that impacts intracellular NAD metabolism and interferes with macrophage cytoskeletal rearrangements subverted by invasive bacteria.


http://ift.tt/2kMW9pR

Endothelial Basement Membrane Laminin 511 Contributes to Endothelial Junctional Tightness and Thereby Inhibits Leukocyte Transmigration

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Jian Song, Xueli Zhang, Konrad Buscher, Ying Wang, Huiyu Wang, Jacopo Di Russo, Lixia Li, Stefan Lütke-Enking, Alexander Zarbock, Anika Stadtmann, Paul Striewski, Benedikt Wirth, Ivan Kuzmanov, Heinz Wiendl, Dörte Schulte, Dietmar Vestweber, Lydia Sorokin
Endothelial basement membranes constitute barriers to extravasating leukocytes during inflammation, a process where laminin isoforms define sites of leukocyte exit; however, how this occurs is poorly understood. In addition to a direct effect on leukocyte transmigration, we show that laminin 511 affects endothelial barrier function by stabilizing VE-cadherin at junctions and downregulating expression of CD99L2, correlating with reduced neutrophil extravasation. Binding of endothelial cells to laminin 511, but not laminin 411 or non-endothelial laminin 111, enhanced transendothelial cell electrical resistance (TEER) and inhibited neutrophil transmigration. Data suggest that endothelial adhesion to laminin 511 via β1 and β3 integrins mediates RhoA-induced VE-cadherin localization to cell-cell borders, and while CD99L2 downregulation requires integrin β1, it is RhoA-independent. Our data demonstrate that molecular information provided by basement membrane laminin 511 affects leukocyte extravasation both directly and indirectly by modulating endothelial barrier properties.

Graphical abstract

Teaser

Endothelial basement membranes are known to restrict leukocyte extravasation; however, how this occurs is unclear. Song et al. show that molecular information gained from basement membrane laminins both directly affects immune cell migration patterns and influences the endothelial barrier properties, thus defining sites where extravasation can occur.


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EBI2 Is Highly Expressed in Multiple Sclerosis Lesions and Promotes Early CNS Migration of Encephalitogenic CD4 T Cells

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Florian Wanke, Sonja Moos, Andrew L. Croxford, André P. Heinen, Stephanie Gräf, Bettina Kalt, Denise Tischner, Juan Zhang, Isabelle Christen, Julia Bruttger, Nir Yogev, Yilang Tang, Morad Zayoud, Nicole Israel, Khalad Karram, Sonja Reißig, Sonja M. Lacher, Christian Reichhold, Ilgiz A. Mufazalov, Avraham Ben-Nun, Tanja Kuhlmann, Nina Wettschureck, Andreas W. Sailer, Klaus Rajewsky, Stefano Casola, Ari Waisman, Florian C. Kurschus
Arrival of encephalitogenic T cells at inflammatory foci represents a critical step in development of experimental autoimmune encephalomyelitis (EAE), the animal model for multiple sclerosis. EBI2 and its ligand, 7α,25-OHC, direct immune cell localization in secondary lymphoid organs. CH25H and CYP7B1 hydroxylate cholesterol to 7α,25-OHC. During EAE, we found increased expression of CH25H by microglia and CYP7B1 by CNS-infiltrating immune cells elevating the ligand concentration in the CNS. Two critical pro-inflammatory cytokines, interleukin-23 (IL-23) and interleukin-1 beta (IL-1β), maintained expression of EBI2 in differentiating Th17 cells. In line with this, EBI2 enhanced early migration of encephalitogenic T cells into the CNS in a transfer EAE model. Nonetheless, EBI2 was dispensable in active EAE. Human Th17 cells do also express EBI2, and EBI2 expressing cells are abundant within multiple sclerosis (MS) white matter lesions. These findings implicate EBI2 as a mediator of CNS autoimmunity and describe mechanistically its contribution to the migration of autoreactive T cells into inflamed organs.

Graphical abstract

Teaser

Wanke et al. show that EBI2 is expressed by Th17 cells in inflammation and that EBI2 promotes early CNS infiltration in passive EAE. Furthermore, they show that CH25H, CYP7B1, and 7α,25-OHC are upregulated in the CNS in EAE and that EBI2 expressing cells are highly present in MS lesions.


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The Atypical Ubiquitin E2 Conjugase UBE2L3 Is an Indirect Caspase-1 Target and Controls IL-1β Secretion by Inflammasomes

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Matthew J.G. Eldridge, Julia Sanchez-Garrido, Gil Ferreira Hoben, Philippa J. Goddard, Avinash R. Shenoy
Caspase-1 activation by inflammasome signaling scaffolds initiates inflammation and antimicrobial responses. Caspase-1 proteolytically converts newly induced pro-interleukin 1 beta (IL-1β) into its mature form and directs its secretion, triggering pyroptosis and release of non-substrate alarmins such as interleukin 1 alpha (IL-1α) and HMGB1. While some caspase-1 substrates involved in these events are known, the identities and roles of non-proteolytic targets remain unknown. Here, we use unbiased proteomics to show that the UBE2L3 ubiquitin conjugase is an indirect target of caspase-1. Caspase-1, but not caspase-4, controls pyroptosis- and ubiquitin-independent proteasomal degradation of UBE2L3 upon canonical and non-canonical inflammasome activation by sterile danger signals and bacterial infection. Mechanistically, UBE2L3 acts post-translationally to promote K48-ubiquitylation and turnover of pro-IL-1β and dampen mature-IL-1β production. UBE2L3 depletion increases pro-IL-1β levels and mature-IL-1β secretion by inflammasomes. These findings regarding UBE2L3 as a molecular rheostat have implications for IL-1-driven pathology in hereditary fever syndromes and in autoinflammatory conditions associated with UBE2L3 polymorphisms.

Graphical abstract

Teaser

Eldridge et al. show that the UBE2L3 ubiquitin E2 conjugating enzyme is targeted for proteasomal degradation by inflammasomes and is therefore an indirect target of human and mouse caspase-1. They also find that UBE2L3 is a post-translational regulator of pro-IL-1β protein levels and thus regulates mature IL-1β production.


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SR-B1 Is a Silica Receptor that Mediates Canonical Inflammasome Activation

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Misato Tsugita, Nobuyuki Morimoto, Manabu Tashiro, Kengo Kinoshita, Masafumi Nakayama
The inhalation of silica dust is associated with fibrosis and lung cancer, which are triggered by macrophage inflammatory responses; however, how macrophages recognize silica remains largely unknown. Here, we identify by functional expression cloning the class B scavenger receptor SR-B1 as a silica receptor. Through an extracellular α-helix, both mouse and human SR-B1 specifically recognized amorphous and crystalline silica, but not titanium dioxide nanoparticles, latex nanoparticles, or monosodium urate crystals, although all particles exhibited negative surface potentials. Genetic deletion of SR-B1 and masking of SR-B1 by monoclonal antibodies showed that SR-B1-mediated recognition of silica is associated with caspase-1-mediated inflammatory responses in mouse macrophages and human peripheral blood monocytes. Furthermore, SR-B1 was involved in silica-induced pulmonary inflammation in mice. These results indicate that SR-B1 is a silica receptor associated with canonical inflammasome activation.

Graphical abstract

Teaser

Recognition of silica by macrophages causes inflammation associated with fibrosis and cancer, but how macrophages recognize silica remains unclear. Tsugita et al. now identify SR-B1 as a silica receptor that mediates canonical inflammasome activation and subsequent lung inflammation.


http://ift.tt/2kMQEr9

Immune Escape via a Transient Gene Expression Program Enables Productive Replication of a Latent Pathogen

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Jessica A. Linderman, Mariko Kobayashi, Vinayak Rayannavar, John J. Fak, Robert B. Darnell, Moses V. Chao, Angus C. Wilson, Ian Mohr
How type I and II interferons prevent periodic reemergence of latent pathogens in tissues of diverse cell types remains unknown. Using homogeneous neuron cultures latently infected with herpes simplex virus 1, we show that extrinsic type I or II interferon acts directly on neurons to induce unique gene expression signatures and inhibit the reactivation-specific burst of viral genome-wide transcription called phase I. Surprisingly, interferons suppressed reactivation only during a limited period early in phase I preceding productive virus growth. Sensitivity to type II interferon was selectively lost if viral ICP0, which normally accumulates later in phase I, was expressed before reactivation. Thus, interferons suppress reactivation by preventing initial expression of latent genomes but are ineffective once phase I viral proteins accumulate, limiting interferon action. This demonstrates that inducible reactivation from latency is only transiently sensitive to interferon. Moreover, it illustrates how latent pathogens escape host immune control to periodically replicate by rapidly deploying an interferon-resistant state.

Graphical abstract

Teaser

How and where interferons suppress latent HSV-1 reactivation is poorly understood. Linderman et al. show that a neuron-intrinsic interferon response blocks reactivation by preventing the initial viral genome-wide burst of transcription. Furthermore, they establish that expressing HSV-1 ICP0 in this first phase temporally restricts IFN action to allow subsequent lytic replication.


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Benzotriazoles Reactivate Latent HIV-1 through Inactivation of STAT5 SUMOylation

Publication date: 31 January 2017
Source:Cell Reports, Volume 18, Issue 5
Author(s): Alberto Bosque, Kyle A. Nilson, Amanda B. Macedo, Adam M. Spivak, Nancie M. Archin, Ryan M. Van Wagoner, Laura J. Martins, Camille L. Novis, Matthew A. Szaniawski, Chris M. Ireland, David M. Margolis, David H. Price, Vicente Planelles
The presence of latent HIV-1 in infected individuals represents a major barrier preventing viral eradication. For that reason, reactivation of latent viruses in the presence of antiretroviral regimens has been proposed as a therapeutic strategy to achieve remission. We screened for small molecules and identified several benzotriazole derivatives with the ability to reactivate latent HIV-1. In the presence of IL-2, benzotriazoles reactivated and reduced the latent reservoir in primary cells, and, remarkably, viral reactivation was achieved without inducing cell proliferation, T cell activation, or cytokine release. Mechanistic studies showed that benzotriazoles block SUMOylation of phosphorylated STAT5, increasing STAT5's activity and occupancy of the HIV-1 LTR. Our results identify benzotriazoles as latency reversing agents and STAT5 signaling and SUMOylation as targets for HIV-1 eradication strategies. These compounds represent a different direction in the search for "shock and kill" therapies.

Graphical abstract

Teaser

Latent HIV-1 represents a barrier toward HIV-1 eradication. Bosque et al. identify a family of compounds that have the ability to reactivate and decrease latent HIV-1. These compounds block SUMOylation of STAT5 and represent a target for HIV-1 eradication strategies.


http://ift.tt/2kQUEWw

A Double-Mediator Based Whole Cell Electrochemical Biosensor for Acute Biotoxicity Assessment of Wastewater

Publication date: Available online 31 January 2017
Source:Talanta
Author(s): Guanyue Gao, Deyu Fang, Yuan Yu, Liangzhuan Wu, Yu Wang, Jinfang Zhi
This work investigates the feasibility and sensitivity of a double-mediator based whole cell electrochemical biosensor to detect the acute biotoxicity of wastewater. The lipophilic mediator menadione was used to mediate the intracellular metabolic activities whereas hydrophilic potassium ferricyanide was employed as extracellular electron acceptor to transport the electron from the menadiol to anode. A chitosan hydrogel polymer film with boron-doped nanocrystalline diamond (BND) particles was electrodeposited onto a glassy carbon (GC) electrode to immobilize Saccharomyces cerevisiae cells and the mediators. The feasibility of the as-prepared biosensor was verified by determine the acute biotoxicity of four heavy metal ions(Cu²⁺, Cd²⁺, Ni²⁺, Pb²⁺), three phenol pollutants (3,5-dichlorophenol, 4-chlorophenol, phenol) and three real wastewater samples. The IC50 values for Cu²⁺, Cd²⁺, Ni²⁺, Pb²⁺ are 10.12mg/L,13.88mg/L, 17.06mg/L and 34.56mg/L. And the IC50 value is 16.48mg/L, 34.40mg/L and 44.55mg/L for 3,5-dichlorophenol, 4-chlorophenol and phenol, respectively. The results of this work indicate that the double-mediator based whole cell electrochemical biosensor could be applied into the acute toxicity assessment of real wastewater samples with excellent performance and highlight their merit as portable and sensitive, which may providing a reasonable and reliable way for wastewater toxicity online detection.

Graphical abstract

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Standardization of Oncoplastic Breast Conserving Surgery

Publication date: Available online 31 January 2017
Source:European Journal of Surgical Oncology (EJSO)
Author(s): Walter P. Weber, Savas Deniz Soysal, Ilario Fulco, Marina Barandun, Doris Babst, Daniel Kalbermatten, Dirk J. Schaefer, Daniel Oertli, Elisabeth A. Kappos, Martin Haug
The emphasis on esthetic outcomes and quality of life after breast cancer surgery has motivated surgeons to develop oncoplastic breast conserving surgery (OPS). Training programs are still rare in most countries, and there is little standardization, which challenges the scientific evaluation of the techniques. The present article attempts to standardize OPS nomenclature, indications, and reconstruction choice selection embedded in a thorough review of the literature. We propose four breast conserving surgery (BCS) categories: Conventional tumorectomy, oncoplastic mastopexy, oncoplastic tumorectomy and oncoplastic reduction mammoplasty. The main volume displacement techniques are glandular re-approximation, use of tailored glandular or dermoglandular flaps and nipple-areola complex pedicles. We developed an indication algorithm based on the size and shape of the breast as well as the size and location of the tumor. A reconstruction algorithm suggests a selection of suitable tailored flaps and pedicles based on tumor location and vascular supply of the breast. The application of these algorithms results in known and novel OPS techniques, which are presented here with long-term results. We designed the algorithms to help tailor every operation to the individual patient in a standardized manner, since OPS is now on the rise, more than two decades after the publication of the first techniques. A rapidly increasing body of observational evidence suggests comparable rates of local recurrence between OPS and conventional BCS. Importantly, the rates of clear resection margins are in favor of OPS despite extended indications to larger tumors. Finally, OPS optimizes patient satisfaction by improving esthetic outcomes after BCS.



http://ift.tt/2kd8HH5

Binding of the chemokine CXCL12α to its natural extracellular matrix ligand heparan sulfate enables myoblast adhesion and facilitates cell motility

Publication date: April 2017
Source:Biomaterials, Volume 123
Author(s): Dhruv Thakar, Fabien Dalonneau, Elisa Migliorini, Hugues Lortat-Jacob, Didier Boturyn, Corinne Albiges-Rizo, Liliane Coche-Guerente, Catherine Picart, Ralf P. Richter
The chemokine CXCL12α is a potent chemoattractant that guides the migration of muscle precursor cells (myoblasts) during myogenesis and muscle regeneration. To study how the molecular presentation of chemokines influences myoblast adhesion and motility, we designed multifunctional biomimetic surfaces as a tuneable signalling platform that enabled the response of myoblasts to selected extracellular cues to be studied in a well-defined environment. Using this platform, we demonstrate that CXCL12α, when presented by its natural extracellular matrix ligand heparan sulfate (HS), enables the adhesion and spreading of myoblasts and facilitates their active migration. In contrast, myoblasts also adhered and spread on CXCL12α that was quasi-irreversibly surface-bound in the absence of HS, but were essentially immotile. Moreover, co-presentation of the cyclic RGD peptide as integrin ligand along with HS-bound CXCL12α led to enhanced spreading and motility, in a way that indicates cooperation between CXCR4 (the CXCL12α receptor) and integrins (the RGD receptors). Our findings reveal the critical role of HS in CXCL12α induced myoblast adhesion and migration. The biomimetic surfaces developed here hold promise for mechanistic studies of cellular responses to different presentations of biomolecules. They may be broadly applicable for dissecting the signalling pathways underlying receptor cross-talks, and thus may guide the development of novel biomaterials that promote highly specific cellular responses.



http://ift.tt/2kbRBZT

Limiting the protein corona: A successful strategy for in vivo active targeting of anti-HER2 nanobody-functionalized nanostars

Publication date: April 2017
Source:Biomaterials, Volume 123
Author(s): Antoine D'Hollander, Hilde Jans, Greetje Vande Velde, Charlotte Verstraete, Sam Massa, Nick Devoogdt, Tim Stakenborg, Serge Muyldermans, Liesbet Lagae, Uwe Himmelreich
Gold nanoparticles hold great promise as anti-cancer theranostic agents against cancer by actively targeting the tumor cells. As this potential has been supported numerously during in vitro experiments, the effective application is hampered by our limited understanding and control of the interactions within complex in vivo biological systems. When these nanoparticles are exposed to a biological environment, their surfaces become covered with proteins and biomolecules, referred to as the protein corona, reducing the active targeting capabilities. We demonstrate a chemical strategy to overcome this issue by reducing the protein corona's thickness by blocking the active groups of the self-assembled monolayer on gold nanostars. An optimal blocking agent, 2-mercapto ethanol, has been selected based on charge and length of the carbon chain. By using a nanobody as a biological ligand of the human epidermal growth factor 2 receptor (HER2), the active targeting is demonstrated in vitro and in vivo in an experimental tumor model by using darkfield microscopy and photoacoustic imaging. In this study, we have established gold nanostars as a conceivable theranostic agent with a specificity for HER2-positive tumors.



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Selective transduction of astrocytic and neuronal CNS subpopulations by lentiviral vectors pseudotyped with Chikungunya virus envelope

Publication date: April 2017
Source:Biomaterials, Volume 123
Author(s): Ioanna Eleftheriadou, Michael Dieringer, Xuan Ying Poh, Julia Sanchez-Garrido, Yunan Gao, Argyro Sgourou, Laura E. Simmons, Nicholas D. Mazarakis
Lentiviral vectors are gene delivery vehicles that integrate into the host genome of dividing and non-dividing mammalian cells facilitating long-term transgene expression. Lentiviral vector versatility is greatly increased by incorporating heterologous viral envelope proteins onto the vector particles instead of the native envelope, conferring on these pseudotyped vectors a modified tropism and host range specificity. We investigated the pseudotyping efficiency of HIV-1 based lentiviral vectors with alphaviral envelope proteins from the Chikungunya Virus (CHIKV-G) and Sindbis Virus (SINV-G). Following vector production optimisation, titres for the CHIKV-G pseudotype were comparable to the VSV-G pseudotype but those for the SINV-G pseudotype were significantly lower. High titre CHIKV-G pseudotyped vector efficiently transduced various human and mouse neural cell lines and normal human astrocytes (NHA) in vitro. Although transduction was broad, tropism for NHAs was observed. In vivo stereotaxic delivery in striatum, thalamus and hippocampus respectively in the adult rat brain revealed localised transduction restricted to striatal astrocytes and hippocampal dentate granule neurons. Transduction of different subtypes of granule neurons from precursor to post-mitotic stages of differentiation was evident in the sub-granular zone and dentate granule cell layer. No significant inflammatory response was observed, but comparable to that of VSV-G pseudotyped lentiviral vectors. Robust long-term expression followed for three months post-transduction along with absence of neuroinflammation, coupled to the selective and unique neuron/glial tropism indicates that these vectors could be useful for modelling and gene therapy studies in the CNS.



http://ift.tt/2jpUB85

Effects of intranasal oxytocin on interpretation and expression of emotions in anorexia nervosa

Abstract

Background

Altered social-emotional functioning is believed to play an important role in the development and maintenance of anorexia nervosa (AN). Recently, there has been increasing interest investigating the role of intranasal oxytocin in social-emotional processing. Thus the present study aimed to investigate the effects of intranasal oxytocin on interpretation and expression of emotions among people with AN.

Method

Thirty women with AN and twenty-nine age-matched healthy women took part in the current study, which used a double blind, placebo controlled, crossover design. The participants received a single dose of 40 IU of intranasal oxytocin in one session and a placebo spray in the other. 15 minutes after administration the participants completed the Reading the Mind in the Eyes Task (RMET) to assess interpretation of complex emotions and mental states followed by a video task, which assessed expressions of facial affect while viewing humorous and sad film clips.

Results

The intranasal oxytocin did not significantly influence expression or interpretation of emotions in the AN or HC groups. The AN group expressed significantly less positive emotion, spent more time looking away, and reported experiencing significantly more negative affect in response to the film clips.

Conclusions

The finding that intranasal oxytocin had little to no effect on interpretation or expression of emotions in either group supports the notion that the effects of oxytocin on social-emotional processing are not straightforward and may depend on individual and environmental differences as well as the emotion being processed. Still, replication of these findings is necessary to explore the effect of timing on the effects of oxytocin before firm conclusions can be drawn. Nonetheless, these findings add to the steady accumulation of evidence that people with AN have reduced emotional expression and avoidance of emotionally provoking stimuli.

This article is protected by copyright. All rights reserved.

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Coffee and tea consumption and risk of leukaemia in an adult population: A reanalysis of the Italian multicentre case-control study

Publication date: April 2017
Source:Cancer Epidemiology, Volume 47
Author(s): Stefano Parodi, Domenico Franco Merlo, Emanuele Stagnaro
BackgroundCoffee and tea are the most frequently consumed beverages in the world. Their potential effect on the risk of developing different types of malignancies has been largely investigated, but studies on leukaemia in adults are scarce.MethodsThe present investigation is aimed at evaluating the potential role of regular coffee and tea intake on the risk of adult leukaemia by reanalysing a large population based case-control study carried out in Italy, a country with a high coffee consumption and a low use of green tea. Interviewed subjects, recruited between 1990 and 1993 in 11 Italian areas, included 1771 controls and 651 leukaemia cases. Association between Acute Myeloid Leukaemia (AML), Acute Lymphoid Leukaemia, Chronic Myeloid Leukaemia, Chronic Lymphoid Leukaemia, and use of coffee and tea was evaluated by standard logistic regression. Odds Ratios (OR) were estimated adjusting for the following potential confounders: gender, age, residence area, smoking habit, educational level, previous chemotherapy treatment, alcohol consumption and exposure to electromagnetic fields, radiation, pesticides and aromatic hydrocarbons.ResultsNo association was observed between regular use of coffee and any type of leukaemia. A small protective effect of tea intake was found among myeloid malignancies, which was more evident among AML (OR=0.68, 95%CI: 0.49–0.94). However, no clear dose-response relation was found.ConclusionThe lower risk of leukaemia among regular coffee consumers, reported by a few of previous small studies, was not confirmed. The protective effect of tea on the AML risk is only partly consistent with results from other investigations.



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Phase 1 study of new formulation of patritumab (U3-1287) Process 2, a fully human anti-HER3 monoclonal antibody in combination with erlotinib in Japanese patients with advanced non-small cell lung cancer

Abstract

Background

This phase 1 study evaluated the safety, tolerability, pharmacokinetics and efficacy of patritumab (U3-1287) Process 2, a new formulation of fully human anti-HER3 monoclonal antibody in combination with erlotinib, an epidermal growth factor receptortyrosine kinase inhibitor (EGFR-TKI) in prior chemotherapy treated Japanese patients with advanced non-small cell lung cancer (NSCLC).

Methods

Patients received intravenous patritumab Process 2 formulation at 9 mg/kg every 3 weeks after initiation of 18 mg/kg loading dose combined with continuous daily dose of erlotinib (150 mg QD) until any of the withdrawal criteria are met. Adverse events (AEs) were assessed using CTCAE v4.0 and tumor response was assessed using RECIST v1.1. Full pharmacokinetic sampling and serum biomarker analyses were mainly performed during cycle 1 and 2.

Results

Total of six EGFR-mutant NSCLC patients including one EGFR-TKI naïve patient received patritumab Process 2 formulation combined with erlotinib. No dose-limiting toxicities were observed. The most frequent AEs were gastrointestinal or skin toxicities, which were generally mild and manageable. One patient discontinued from study due to reversible grade 3 interstitial lung disease. The mean area under the curve (AUC) value was 2640 μg/day/mL; the Cmax value was 434 μg/mL, respectively. The median progression-free survival (95% confidence interval) was 220.0 (100.0–363.0) days. HER3 ligand heregulin was detected in serum from only a patient that maintained most durable stable disease.

Conclusions

Patritumab Process 2 formulation in combination with erlotinib was well tolerated compatible with favorable PK profile in Japanese patients with advanced NSCLC.

http://ift.tt/2kcDp3a

Morphology of palatally impacted canines: A case-controlled cone-beam volumetric tomography study

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Pilana Vithanage Kalani Shihanika Hettiarachchi, Richard John Olive, Paul Monsour
IntroductionThe aim of this study was to investigate the relationships between an apical curvature or a hook and the crown/root ratio in subjects with and without palatally impacted maxillary canines.MethodsAn experimental group of 44 patients (17 boys, 27 girls; mean age, 13.6 years) with 59 palatally impacted maxillary canines was selected from the records of patients referred to a radiology practice for cone-beam imaging. If a patient had bilateral palatally impacted canines, 1 canine was randomly selected for analysis. The palatally impacted canine group was matched for age and sex with 49 normal subjects (25 boys, 24 girls; mean age, 13.2 years) with 98 canines. Cone-beam DICOM files were imported into In Vivo imaging software (version 5.3; Anatomage, San Jose, Calif) for analysis. The angulations and linear variables of the maxillary canines were measured by using the software measurement tools. Chi-square and independent t tests were used to test for differences between the groups.ResultsThe presence of a hook at the apical third and other root curvature were significantly different between the 2 groups (P <0.001 and P <0.05, respectively). Of the 44 palatally impacted canines, 16 (36.4%) had an apical hook and only 1 canine in the control group had an apical hook (1.0%). The mean root length of the palatally impacted canines was 2.66 mm shorter (P <0.001), and the mean crown/root ratio was significantly greater for the palatally impacted canines compared with the nonimpacted group (P <0.001).ConclusionsPalatally impacted canines have a greater tendency to develop apical hooks and are less likely to develop other root curvatures than are nonimpacted canines. Also, they have shorter roots resulting in larger crown/root ratios compared with the control group.



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Editorial Board

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2





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February 2017:151(2)

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Allen H. Moffitt




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Information for readers

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2





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Association between 3-dimensional mandibular morphology and condylar movement in subjects with mandibular asymmetry

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Michiyo Ikeda, Jun J. Miyamoto, Jun-ichi Takada, Keiji Moriyama
IntroductionThe purpose of this study was to evaluate the hypothesis that 3-dimensional mandibular morphology is correlated with condylar movement in patients with mandibular asymmetry.MethodsSubjects were classified into 2 groups (n = 25 each): mandibular asymmetry with a menton deviation greater than 4 mm and no mandibular asymmetry with a menton deviation less than 4 mm. Linear and volumetric measurements of 3-dimensional mandibular morphology were recorded using computed tomography. Mandibular functional movement was recorded by computerized axiography (CADIAX; Gamma Dental, Klosterneuburg, Austria), and condylar path length, sagittal condylar inclination, and transverse condylar inclination on protrusion were measured. We calculated side-to-side asymmetry (shifted side vs nonshifted side) in mandibular morphology and assessed condylar movement by using an asymmetry ratio (nonshifted side/shifted side).ResultsSignificant differences in mandibular morphology and condylar movement were found between the 2 groups. In the group with menton deviation greater than 4 mm, significant correlations were found between the asymmetry ratio of mandibular morphology and condylar movement: ie, condylar path length and transverse condylar inclination. No significant correlations were found between any of these measurements in the group with menton deviation less than 4 mm.ConclusionsIn support of our hypothesis, the results suggested that 3-dimensional mandibular morphologic asymmetry is associated with condylar movement in subjects with mandibular asymmetry.



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A new definition for oral health developed by the FDI World Dental Federation opens the door to a universal definition of oral health

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Michael Glick, David M. Williams, Dushanka V. Kleinman, Marko Vujicic, Richard G. Watt, Robert J. Weyant




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Root resorption of maxillary incisors retracted with and without skeletal anchorage

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Sérgio Estelita Barros, Guilherme Janson, Kelly Chiqueto, Vitor Oliveira Baldo, Taiana Oliveira Baldo
IntroductionOur objective was to compare root resorption degree of the maxillary central incisors retracted with and without skeletal anchorage.MethodsThis nonrandomized historical control study included 37 patients requiring maximum anterior retraction and treated with extraction of 2 maxillary premolars. Group 1 consisted of 22 patients (11 male, 11 female) in whom anterior retraction was performed without skeletal anchorage, and group 2 included 15 patients (3 male, 12 female) treated with skeletally anchored anterior retraction. Periapical radiographs were used to evaluate root resorption degree by a scoring system. The groups were compared regarding the resorption score and resorption degree distribution with the Mann-Whitney U test, chi-square test, and Z test on proportions.ResultsThere was no statistically significant intergroup difference regarding root resorption, but the number of patients with severe and extreme root resorption degrees was significantly greater in group 2.ConclusionsAlthough the root resorption degree of the skeletal anchorage group was not significantly different from the group without skeletal anchorage, the number of patients with severe to extreme resorption in the first group was significantly greater. Therefore, careful clinical monitoring of skeletally anchored anterior retraction is needed, especially when there are known root resorption predisposing factors.



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Rationale behind Twin-block incline

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Michael J. Trenouth




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Authors' response

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Peilun Li, Jing Feng, Gang Shen, Ning Zhao




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Table of Contents

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2





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Indirectly loaded miniscrews for anchorage control

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Xiaolong Li, Wenli Lai




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Growth changes in the anterior and middle cranial bases assessed with cone-beam computed tomography in adolescents

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Mona Afrand, Heesoo Oh, Carlos Flores-Mir, Manuel O. Lagravère-Vich
IntroductionInitially, cone-beam computed tomography images from dry skulls were used to 3 dimensionally evaluate intrarater and interrater reliabilities and accuracy of selected 3-dimensional landmarks located in the anterior and middle cranial bases. Thereafter, dimensional changes of the anterior and middle cranial bases with growth were evaluated by using the previously selected landmarks.MethodsCone-beam computed tomography images of 10 dry skulls were used to identify useful landmarks from different areas of the anterior and middle cranial bases based on their reliability and accuracy. These selected landmarks were identified in the images of an already available sample of adolescents (n = 60) taken at 2 time points (19 months apart) to assess dimensional changes with growth.ResultsThe majority of the proposed 3-dimensional landmarks with the exception of the lesser wing of the sphenoid showed acceptable intrarater and interrater reliabilities. The distances measured between foramina and canals in the transverse dimension showed evidence of increases in size. However, the mean amounts of increase in these transverse distances were equal to or less than 1.05 mm (from 1.1% to 4.1%). No change was observed between the right and left anterior and posterior clinoid processes. The vertical dimensions showed evidence of some changes, but these were within 2% of the original distances.ConclusionsIn this adolescent sample, minor growth-related changes were observed in the anterior and middle cranial bases. The midsagittal area of the anterior cranial base (foramen caecum to presphenoid) was stable. The right and left anterior and posterior clinoid processes can be used for transverse superimposition.



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Authors' response

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Nitika Monga, Om Prakash Kharbanda, Vilas Samrit




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Transport distraction osteogenesis combined with orthodontic treatment in a patient with unilateral temporomandibular joint ankylosis

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Seo-Yeon Jung, Jung-Hyun Park, Hyung-Sik Park, Hyoung-Seon Baik
Temporomandibular joint ankylosis manifests a range of clinical characteristics dependent upon the age of onset, the affected side, and the severity. When it occurs during development, mandibular growth is affected, resulting in severe retrusion or asymmetry as well as limitation of mandibular movements. Progressive asymmetric mandibular growth in unilateral ankylosis causes canting of the occlusal plane. In this case report, we present a successful temporomandibular joint reconstruction using transport distraction osteogenesis combined with camouflage orthodontic treatment for occlusal canting correction of a patient with unilateral temporomandibular joint ankylosis and severe facial asymmetry.



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More precision in expansion, more accuracy in retention

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Thiago L. Monte, Paulo Roberto Aranha Nouer




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When communicating isn't enough

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Laurance Jerrold




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Authors' response

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): José Valladares-Neto, Karine Evangelista, Hianne Miranda de Torres, Matheus Melo Pithon, Maria Alves Garcia Santos Silva




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Directory: AAO Officers and Organizations

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2





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Esthetic evaluation of implants vs canine substitution

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Katja Kritzler




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Authors’ response

Publication date: February 2017
Source:American Journal of Orthodontics and Dentofacial Orthopedics, Volume 151, Issue 2
Author(s): Ute Schneider, Lorenz Moser, Marzia Fornasetti, Michele Piattella, Giuseppe Siciliani




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Standards for Reporting EMG Data

Publication date: February 2017
Source:Journal of Electromyography and Kinesiology, Volume 32





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Debaryomyces hansenii up regulates superoxide dismutase gene expression and enhances the immune response and survival in Pacific red snapper (Lutjanus peru) leukocytes after Vibrio parahaemolyticus infection

Publication date: June 2017
Source:Developmental & Comparative Immunology, Volume 71
Author(s): Carlos Angulo, Minerva Maldonado, Karen Delgado, Martha Reyes-Becerril
Application of yeast is increasing to improve welfare and promotes growth in aquaculture. The halotolerant yeast Debaryomyces hansenii is normally a non-pathogenic yeast with probiotic properties and potential source of antioxidant enzymes as superoxide dismutase. Here, first, we characterized the sequence features of MnSOD and icCu/ZnSOD from Pacific red snapper, and second, we evaluated the potential antioxidant immune responses of the marine yeast Debaryomyces hansenii strain CBS004 in leukocytes which were then subjected to Vibrio parahaemolyticus infection. In silico analysis revealed that LpMnSOD consisted of 1186 bp, with an ORF of 678 bp encoding a 225 amino acid protein and LpicCu/ZnSOD consisted of 1090 bp in length with an ORF of 465 bp encoding a 154 amino acid protein. Multiple alignment analyzes revealed many conserved regions and active sites among its orthologs. In vitro assays using head-kidney and spleen leukocytes immunostimulated with D. hansenii and zymosan in response to V. parahaemolyticus infection reveled that D. hansenii strain CBS004 significantly increased transcriptions of MnSOD and icCu/ZnSOD genes. Flow cytometry assay showed that D. hansenii was able to inhibit apoptosis caused by V. parahaemolyticus in the Pacific red snapper leukocytes and enhanced the phagocytic capacity in head-kidney leukocytes. Immunological assays reveled an increased in superoxide dismutase and peroxidase activities, as well as, in nitric oxide production and reactive oxygen species production (respiratory burst) in fish stimulated with D. hansenii. Finally, our results. These results strongly support the idea that marine yeast Debaryomyces hansenii strain CBS004 can stimulate the antioxidant immune mechanism in head-kidney and spleen leukocytes.



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Inside front cover

Publication date: 1 April 2017
Source:Talanta, Volume 165





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A feasible method for indirect quantification of L-T4 in drugs by iodine determination

Publication date: 1 May 2017
Source:Talanta, Volume 166
Author(s): Marcia Foster Mesko, Alessandra Cortes Teotonio, Dirce Taina Teixeira Oliveira, Diogo La Rosa Novo, Vanize Caldeira Costa
In this work, a method combining microwave-induced combustion (MIC) for sample preparation of commercial levothyroxine sodium (L-T4) drugs (L-T4: 25−200µg/tablet), and potentiometry with ion selective electrode (ISE) for iodine determination and subsequent indirect quantification of L-T4 was proposed. The type and concentration of the absorbing solution were evaluated to select the most suitable conditions for this study. Using the MIC method, it was possible to use solutions as diluted as 150mmolL⁻¹ (NH4)2CO3 (for samples containing 25−200µg of L-T4/tablet) for I absorption. In these conditions, recoveries for L-T4 were between 99% and 101%, and relative standard deviations were lower than 10%. The limit of detection for L-T4 was 11.2µg/tablet, which is almost two times lower than the minimum concentration of L-T4 in commercially available drugs. Thus, the MIC was suitable for the digestion of several L-T4 drugs for subsequent I determination by ISE and indirect quantification of L-T4. Furthermore, the proposed method presents high throughput with low reagent consumption and consequently lower waste generation, making it suitable for routine determination of L-T4 in drugs. From the obtained results, it was possible to observe that one of the analyzed samples is not in agreement with the limits established by the United States Pharmacopeia, indicating the importance of the drug quality control. The United States Pharmacopeia establishes that each tablet must contain between 90% and 110% of the amount of active substance declared by the manufacturer.

Graphical abstract

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Smartphone-based immunosensor for CA125 detection

Publication date: 1 May 2017
Source:Talanta, Volume 166
Author(s): Oana Hosu, Andrea Ravalli, Giuseppe Mattia Lo Piccolo, Cecilia Cristea, Robert Sandulescu, Giovanna Marrazza
In this work, we report the design, the development and the characterization of the analytical performances of a colorimetric smartphone-based immunosensor for the detection of cancer antigen 125 (CA125). The immunosensor was based on a sandwich strategy in which the primary antibody was immobilized by spotting onto the 3D nitrocellulose membrane. The immunospots were subsequently incubated with CA125 solutions, followed by the affinity reaction with a secondary antibody labeled with gold nanoparticles (AuNPs). The antibody-AuNPs captured onto immunospots induced the silver deposition from a silver enhancer solution leading to the formation of gold-silver nanoparticles of different grey color spots depending on CA125 concentration. The 8 megapixels smartphone camera was integrated in a home-made dark box and used as transducer of color image acquisition and data handling. The pixel intensity of the captured images was determined by an image processing algorithm. The experimental parameters involved in each step of the immunosensor design were studied and optimized, obtaining a limit of detection of 30U/mL CA125. The selectivity of the immunoassay was proven against different concentration solutions of Vascular Endothelial Growth Factor (VEGF) antigen as an unspecific protein when a blank signal was obtained for all tested solutions. Finally, preliminary experiments in human serum samples spiked with CA125 protein were also performed. Therefore, the proposed system could represent a powerful point-of-care tool for the next generation technology for detecting and monitoring cancer biomarkers at early stages by taking advantage of nowadays gadgets with enhanced features such as smartphones.

Graphical abstract

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A single molecular fluorescent probe for selective and sensitive detection of nitroaromatic explosives: A new strategy for the mask-free discrimination of TNT and TNP within same sample

Publication date: 1 May 2017
Source:Talanta, Volume 166
Author(s): Zhe Zhang, Shusen Chen, Rui Shi, Jiawen Ji, Dequan Wang, Shaohua Jin, Tongyu Han, Chenxiao Zhou, Qinghai Shu
A simple naphthalene based fluorescent probe was first time reported as dual sensing of 2,4,6-trinitrotolune (TNT) and 2,4,6-trinitrophenol (TNP) by distinguishable changes in both solution color change and fluorescence within same sample without any mask agent. Upon addition of TNT and TNP, the strong emission quenching at 412nm and a new emission band at 530nm was observed, respectively. In addition, the sensing mechanism was evaluated by DFT calculations by Gaussian 09 software.

Graphical abstract

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Fast method to quantify PAHs in contaminated soils by direct thermodesorption using analytical pyrolysis

Publication date: 1 May 2017
Source:Talanta, Volume 166
Author(s): C. Biache, C. Lorgeoux, A. Saada, S. Colombano, P. Faure
A method for polycyclic aromatic hydrocarbon (PAH) quantification, based on pyrolysis at 450°C combined with gas chromatography coupled with mass spectrometry and flame ionization detection (Py-GC-MS/FID), was developed and compared to a conventional PAH quantification method using accelerated solvent extraction and GC–MS analyses. The PAH contents of three coking plant soils, one gas plant soil, two wood-treating facility soils and one certified reference material (CRM - BCR 524) were determined using both methods. The results obtained with both methods showed a good match, especially in the case of the CRM. The other soil samples presented higher variability which was greatly reduced by crushing the samples to lower particle size (from <500 to <100µm). Higher contents of low molecular weight (LMW) PAHs were quantified with the Py-GC-MS/FID than with the conventional method, probably because of a slight cracking phenomenon occurring during the pyrolysis and/or a loss of the LMW compounds during the sample concentration required for the conventional method. Because of the limited sample preparation and the fact that no solvent was used, the pyrolysis-based method was proven to be a faster, less expensive and more environmentally friendly than the classical methods for PAH quantification in contaminated soils.

Graphical abstract

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Editorial Board

Publication date: 1 April 2017
Source:Talanta, Volume 165





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Antagonizing the different stages of kappa opioid receptor activation selectively and independently attenuates acquisition and consolidation of associative memories

Publication date: 14 April 2017
Source:Behavioural Brain Research, Volume 323
Author(s): Ryan Loh, Lily Chau, Ali Aijaz, Kevin Wu, Roberto Galvez
Previous work from our laboratory has shown that nonspecific kappa opioid receptor (KOR) antagonism in primary somatosensory cortex (S1) can inhibit acquisition for the forebrain-dependent associative task, Whisker-Trace Eyeblink conditioning (WTEB). Although studies have demonstrated that KOR activation can alter stimuli salience, our studies controlled for these factors, demonstrating that KOR also plays a role in facilitating learning. KOR has two distinct phases of activation followed by internalization/downregulation, that each independently activate kinases and transcription factors known to mediate task acquisition and memory consolidation respectively. The current study demonstrated that antagonism of the initial phase of KOR activation in S1 via local injections of the g-protein inhibitor, pertussis toxin (PTX), blocked initial WTEB acquisition without affecting retention of the association. In contrast, KOR late phase antagonism in S1 via local injections of the GRK3-specific antagonist, guanidinonaltrindole (GNTI), blocked retention of the WTEB association without affecting task acquisition. Consistent with the known mechanism for KOR activation, KOR protein expression in S1 was found to be decreased following WTEB training, further supporting the involvement of neocortical KOR activation with learning. Prior studies have shown that task acquisition and memory consolidation are mediated by distinct molecular processes; however, little is known regarding a potential mechanism driving these processes. The current study suggests that neocortical KOR activation mediates activation of these processes with learning. This study provides the first evidence for a time- and learning-dependent property of neocortical KOR in facilitating acquisition and consolidation of associative memories, while elucidating an unexplored neocortical learning mechanism.



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Nrf2- and Bach1 May Play a Role in the Modulation of Ultraviolet A-Induced Oxidative Stress by Acetyl-11-Keto-β-Boswellic Acid in Skin Keratinocytes

Background: Exposure of human skin to solar ultraviolet A (UVA) irradiation causes severe oxidative stress with damage to various cellular components and concomitant inflammation and carcinogenesis. Objective: The aim of this study is to investigate the protective effect of acetyl-11-keto-β-boswellic acid (AKBA) against UVA radiation on human skin keratinocytes. Methods: HaCaT cells were pretreated with AKBA followed by UVA irradiation. Radiation effects on cell morphology, cell viability, intracellular reactive oxygen species (ROS) levels, and antioxidant enzymes were examined. Results: AKBA reduces UVA irradiation-induced cell viability loss, accompanied by a decreased production of UVA-induced ROS, decreased malondialdehyde, and increased superoxide dismutase expression. In addition, AKBA increased basal and UVA-induced levels of Nrf2 (NF-E2-related factor 2), the redox-sensitive factor, and its target genes NQO1 and heme oxygenase-1 (HO-1), whereas expression of the transcriptional repressor Bach1 (BTB and CNC homology 1) was reduced. Furthermore, the cytoprotective effects of AKBA against UVA-derived oxidative damage were accompanied by modulating expression of inflammatory mediators (i.e., cyclooxygenase-2 and nuclear factor-#x03BA;B) and NOX1. Conclusions: AKBA protects skin cells from UVA-induced damage by modulating inflammatory mediators and/or ROS production. Therefore, AKBA has potential in the development of skin care products.
Skin Pharmacol Physiol 2017;30:13-23

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Intact wound repair activity of human mesenchymal stem cells after YM155 mediated selective ablation of undifferentiated human embryonic stem cells

Publication date: Available online 31 January 2017
Source:Journal of Dermatological Science
Author(s): Keun-Tae Kim, Ho-Chang Jeong, C-Yoon Kim, Eun-Young Kim, Si-Hyun Heo, Seung-Ju Cho, Ki-Sung Hong, Hyuk-Jin Cha
BackgroundRisk of teratoma formation during human pluripotent stem cell (hPSC)-based cell therapy is one of the technical hurdles that must be resolved before their wider clinical application. To this end, selective ablation of undifferentiated hPSCs has been achieved using small molecules whose application should be safe for differentiated cells derived from the hPSCs.ObjectiveHowever, the functional safety of such small molecules in the cells differentiated from hPSCs has not yet been extensively validated.MethodWe used the survivin inhibitor YM155, which induced highly selective cell death of hPSCs for ablating undifferentiated hESCs after differentiation to human mesenchymal stem cells (hMSCs) and examined whether hMSCs remained fully functional after being exposed by YM155.ResultsWe demonstrated that human mesenchymal stem cells (hMSCs) derived from human embryonic stem cells (hESCs) remained fully functional in vitro and in vivo, while hESCs were selectively ablated.ConclusionThese results suggest that a single treatment with YM155 after differentiation of hMSCs would be a valid approach for teratoma-free cell therapy.



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Perirhinal cortex tracks degree of recent as well as cumulative lifetime experience with object concepts

Publication date: Available online 31 January 2017
Source:Cortex
Author(s): Devin Duke, Chris B. Martin, Ben Bowles, Ken McRae, Stefan Köhler
Evidence from numerous sources indicates that recognition of the prior occurrence of objects requires computations of perirhinal cortex (PrC) in the medial temporal lobe. Extant research has primarily probed recognition memory based on item exposure in a recent experimental study episode. Outside the laboratory, however, familiarity for objects typically accrues gradually with learning across many different episodic contexts, which can be distributed over a lifetime of experience. It is currently unknown whether PrC also tracks this cumulative lifetime experience with object concepts accrued outside the laboratory. To address this issue, we conducted an fMRI experiment in healthy individuals in which we compared judgments of the perceived lifetime familiarity with object concepts, a task that has previously been employed in many normative studies on concept knowledge, with frequency judgments for recent laboratory exposure in a study phase. Guided by neurophysiological data showing that neurons in primate PrC signal prior object exposure at multiple time scales, we predicted that PrC responses would track perceived prior experience in both types of judgments. Left perirhinal cortex and a number of cortical regions that are often co-activated as part of the default-mode network showed an increase in BOLD response in relation to increases in the perceived cumulative lifetime familiarity of object concepts. These regions included the left hippocampus, left mid-lateral temporal cortex, as well as anterior and posterior cortical midline structures. Critically, left PrC was found to be the only region that showed this response in combination with the typically observed decrease in signal for perceived recent exposure in the experimental study phase. These findings provide, to our knowledge, the first evidence tying signals in human PrC to variations in cumulative lifetime experience with object concepts. These results offer a new link between its role in recognition memory and a broader role in conceptual processing.



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Action perception as hypothesis testing

Publication date: Available online 31 January 2017
Source:Cortex
Author(s): Francesco Donnarumma, Marcello Costantini, Ettore Ambrosini, Karl Friston, Giovanni Pezzulo
We present a novel computational model that describes action perception as an active inferential process that combines motor prediction (the reuse of our own motor system to predict perceived movements) and hypothesis testing (the use of eye movements to disambiguate amongst hypotheses). The system uses a generative model of how (arm and hand) actions are performed to generate hypothesis-specific visual predictions, and directs saccades to the most informative places of the visual scene to test these predictions – and underlying hypotheses. We test the model using eye movement data from a human action observation study. In both the human study and our model, saccades are proactive whenever context affords accurate action prediction; but uncertainty induces a more reactive gaze strategy, via tracking the observed movements. Our model offers a novel perspective on action observation that highlights its active nature based on prediction dynamics and hypothesis testing.



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Antioxidative and anticancer properties of Licochalcone A from licorice

Publication date: 23 February 2017
Source:Journal of Ethnopharmacology, Volume 198
Author(s): Xiangrong Chen, Zuojia Liu, Rizeng Meng, Ce Shi, Na Guo
Ethnopharmacological relevanceLicochalcone A (LCA) is a characteristic chalcone that is found in licorice, which is a traditional medicinal plant. In traditional medicine, LCA possesses many potential biological activities, including anti-parasitic, anti-inflammatory and antitumor activities.Aim of the studyTo determine the antioxidant activity of LCA and, on this basis, to investigate the role of its anticancer activity.Materials and methodsTo validate the antioxidant activity of LCA, the proteins SOD, CAT and GPx1 were analyzed using western blotting and cellular antioxidant activity (CAA) assays. Oxidative free radicals are associated with cancer cells. Therefore, the anticancer activity of LCA was also evaluated. To assess the anticancer activity, cell viability assays were performed and apoptosis was evaluated. In addition, MAPK-related proteins were analyzed using western blotting.ResultsThe experimental data showed that the EC50 of LCA is 58.79±0.05μg/mL and 46.29±0.05μg/mL under the two conditions tested, with or without PBS. In addition, LCA at a concentration of approximately 2–8μg/mL can induce the expression of SOD, CAT and GPx1 proteins. Further, LCA inhibits the growth of HepG2 cells through cell proliferation arrest and the subsequent induction of apoptosis, and LCA attenuated the p38/JNK/ERK signaling pathway in a dose-dependent manner.ConclusionThe results showed that LCA suppresses the oxidation of cells and markedly inhibits the proliferation of cancer cells. These findings confirm the traditional use of LCA in folk medicine.

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Anti-malarial synergy of secondary metabolites from Morinda lucida Benth

Publication date: Available online 30 January 2017
Source:Journal of Ethnopharmacology
Author(s): Bertha Chithambo, Xavier Siwe Noundou, Rui W.M. Krause
Ethnopharmacological relevanceThe roots, stem and leaves of Morinda lucida are used in some African countries as treatment against different types of fevers including yellow fever, malaria, trypanosomiasis and feverish conditions during child birth.Aim of the studyTo determine the in vitro cell toxicity and anti-malarial activity of the extracts of stem bark of M. lucida and to identify the secondary metabolites in the extract that may be responsible for this activity.Materials and methodsThe cell toxicity studies of crude extract [dichloromethane (DCM):Methanol (MeOH) in a ratio of1:1 (v/v)] as well as compounds isolated from the same extract were carried out using human cervix adenocarcinoma cells (HeLa cells); while the anti-malarial activities of the same samples were performed against Plasmodium falciparum strain 3D7 using the parasite lactate dehydrogenase (pLDH) assay. The isolation of the active compounds was carried out using chromatographic techniques (column and thin layer chromatography) where as mass spectrometry (MS), Fourier transform infrared spectroscopy (FTIR) as well as 1D- and 2D- nuclear magnetic resonance (NMR) analyses were employed in the characterisation and identification of the isolated secondary metabolites.ResultsThe pLDH and cell toxicity assays for the crude extract and the fractions of M. lucida indicated that some fractions reduced the malaria parasite viability by approximately 50% at 100μg/mL and they were not significantly cytotoxic. An IC50 done on the crude extract gave a value of 25μg/mL. The % cell viability for the crude extract in cell toxicity assay remained at 100%. Seven chemical constituents i.e. asperuloside (1), asperulosidic acid (2), stigmasterol (3a), β-sitosterol (3b), cycloartenol (3c), campesterol (3d) and 5,15-O-dimethylmorindol (4) were isolated from the DCM-MeOH extract of stem bark. The isolated compounds tested were not that active by themselves individually at 20μM but their activities were increased when the isolated compounds were combined. As seen when compounds 2, 3 and 4 (% viability: 93, 123 and 101 respectively) were combined yielding an IC50 value of 17μM. Furthermore, this is the first report of compounds 1, 2, 3c, 3d and 4 isolated from M. lucida.ConclusionThe crude extract completely suppressed the growth of P. falciparum. This indicates that the crude extract contains many compounds that might be acting in synergy. The observed activity of the crude extract and the samples containing a mixture of different compounds support the traditional use of M. lucida for the treatment of malaria.

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