bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Botrytis cinerea elicitor protein BcIEB1 interacts with the tobacco PR5‐family protein osmotin and protects the fungus against its antifungal activity
Authors: González Mario , Brito Nélida , González Celedonio 
Published: May 2017

Summary

The broad‐range phytopathogenic fungus Botrytis cinerea secretes hundreds of proteins during infection of its plant hosts. One of these proteins, BcIEB1, is abundantly secreted and is able to elicit plant defenses, probably as a pathogen‐associated molecular pattern, although its native function in B. cinerea biology remains unknown.

Pull‐down experiments designed to isolate the molecular target of BcIEB1 in tobacco resulted in the identification of osmotin, a pathogenesis‐related protein of family 5 that shows antifungal activity. The expression of osmotin in Escherichia coli allowed the verification of the BcIEB1–osmotin interaction with pure proteins by pull‐down and far Western blot experiments, as well as the confirmation of the activity of osmotin against B. cinerea.

Interestingly, B. cinerea Δbcieb1 mutants are more susceptible than the wild‐type to osmotin, and the external addition of pure BcIEB1 protects the Δbcieb1 mutants, as well as Saccharomyces cerevisiae, from the antifungal action of osmotin, thus pointing at PR5 inhibition as the primary native function of BcIEB1.

The question of whether osmotin is also involved in the activation of plant defenses by BcIEB1 is also addressed, and the data suggest that osmotin does not participate in the elicitation process.
Arabidopsis ketoacyl‐CoA synthase 16 (KCS16) forms C36/C38 acyl precursors for leaf trichome and pavement surface wax
Authors: Hegebarth Daniela , Buschhaus Christopher , Joubès Jérôme , Thoraval Didier , Bird David , Jetter Reinhard 
Published: May 2017

AbstractThe aliphatic waxes sealing plant surfaces against environmental stress are generated by fatty acid elongase complexes, each containing a β‐ketoacyl‐CoA synthase (KCS) enzyme that catalyses a crucial condensation forming a new C─C bond to extend the carbon backbone. The relatively high abundance of C35 and C37 alkanes derived from C36 and C38 acyl‐CoAs in Arabidopsis leaf trichomes (relative to other epidermis cells) suggests differences in the elongation machineries of different epidermis cell types, possibly involving KCS16, a condensing enzyme expressed preferentially in trichomes. Here, KCS16 was found expressed primarily in Arabidopsis rosette leaves, flowers and siliques, and the corresponding protein was localized to the endoplasmic reticulum. The cuticular waxes on young leaves and isolated leaf trichomes of ksc16 loss‐of‐function mutants were depleted of C35 and C37 alkanes and alkenes, whereas expression of Arabidopsis KCS16 in yeast and ectopic overexpression in Arabidopsis resulted in accumulation of C36 and C38 fatty acid products. Taken together, our results show that KCS16 is the sole enzyme catalysing the elongation of C34 to C38 acyl‐CoAs in Arabidopsis leaf trichomes and that it contributes to the formation of extra‐long compounds in adjacent pavement cells.
Role of sex in morbidity and mortality of very premature neonates
Authors: Ito Masato , Tamura Masanori , Namba Fumihiko ,   the Neonatal Research Network of Japan
Published: May 2017

AbstractBackgroundThe aim of this study was to investigate the effect of sex on survival and short‐term outcomes of very low‐birthweight infants (VLBWI) born in Japan.MethodsThis study included VLBWI who were admitted to neonatal intensive care units participating in the Neonatal Research Network of Japan, between 2003 and 2012. The primary outcome was a composite of mortality or any major morbidity, including neurologic injury, bronchopulmonary dysplasia (BPD), necrotizing enterocolitis (NEC), or retinopathy of prematurity requiring treatment.ResultsThe composite primary outcome was worse in male infants. Male infants were also more likely to develop BPD. This difference reached statistical significance in neonates born at ≥26 weeks of gestation. In contrast, significant difference in the incidence of NEC was more likely to be seen at 23–25 weeks of gestation.ConclusionMale neonates are more likely to die and are at a higher risk of respiratory and gastrointestinal complications than female neonates.
Tolyporphin—An Unusual Green Chlorin‐like Dioxobacteriochlorin
Authors: Brückner Christian 
Published: May 2017

AbstractThe tolyporphins, a family of green tetrapyrrolic pigments isolated from a cyanobacterium‐microbial ecological unit, possess unique carbohydrate‐derivatized dioxobacteriochlorin frameworks. A brief overview over the history, synthesis, chemistry and biological properties of the tolyporphins forms the backdrop for highlighting the contribution by Hood et al. (Photochem. Photobiol., 2017, https://doi.org/10.1111/php.12781) who demonstrate the optical properties of tolyporphins to be more similar to those of chlorins than of bacteriochlorins. This property could be correlated with the presence of β‐oxo‐functionalities. The study continues to clarify the structure, properties and possible roles of these intriguing chromophores with a range of biomedical properties.
Volitional saccade performance in a large sample of patients with obsessive‐compulsive disorder and unaffected first‐degree relatives
Authors: Bey Katharina , Kloft Lisa , Lennertz Leonhard , Grützmann Rosa , Heinzel Stephan , Kaufmann Christian , Klawohn Julia , Riesel Anja , Meyhöfer Inga , Kathmann Norbert , Wagner Michael 
Published: May 2017

AbstractRecent evidence indicates that patients with obsessive‐compulsive disorder (OCD) as well as their unaffected first‐degree relatives show deficits in the volitional control of saccades, suggesting that volitional saccade performance may constitute an endophenotype of OCD. Here, we aimed to replicate and extend these findings in a large, independent sample. One hundred and fifteen patients with OCD, 103 healthy comparison subjects without a family history of OCD, and 31 unaffected first‐degree relatives of OCD patients were examined using structured clinical interviews and performed a volitional saccade task as well as a prosaccade task. In contrast to previous reports, neither patients nor relatives showed impairments in the performance of volitional saccades compared to healthy controls. Notably, medicated patients did not differ from nonmedicated patients, and there was no effect of depressive comorbidity. Additional analyses investigating correlations between saccade performance and OCD symptom dimensions yielded no significant associations. In conclusion, the present results do not support the notion that volitional saccade execution constitutes an endophenotype of OCD. Possible explanations for inconsistencies with previous studies are discussed.
The brassinosteroid receptor BRI1 can generate cGMP enabling cGMP‐dependent downstream signaling
Authors: Wheeler Janet I. , Wong Aloysius , Marondedze Claudius , Groen Arnoud J. , Kwezi Lusisizwe , Freihat Lubna , Vyas Jignesh , Raji Misjudeen A. , Irving Helen R. , Gehring Chris 
Published: May 2017

SummaryThe brassinosteroid receptor brassinosteroid insensitive 1 (BRI1) is a member of the leucine‐rich repeat receptor‐like kinase family. The intracellular kinase domain of BRI1 is an active kinase and also encapsulates a guanylate cyclase catalytic centre. Using liquid chromatography tandem mass spectrometry, we confirmed that the recombinant cytoplasmic domain of BRI1 generates pmol amounts of cGMP per μg protein with a preference for magnesium over manganese as a co‐factor. Importantly, a functional BRI1 kinase is essential for optimal cGMP generation. Therefore, the guanylate cyclase activity of BRI1 is modulated by the kinase while cGMP, the product of the guanylate cyclase, in turn inhibits BRI1 kinase activity. Furthermore, we show using Arabidopsis root cell cultures that cGMP rapidly potentiates phosphorylation of the downstream substrate brassinosteroid signaling kinase 1 (BSK1). Taken together, our results suggest that cGMP acts as a modulator that enhances downstream signaling while dampening signal generation from the receptor.
Expression and targeting of transcription factor ATF5 in dog gliomas
Authors: York D. , Sproul C. D. , Chikere N. , Dickinson P. J. , Angelastro J. M. 
Published: May 2017

BackgroundActivating transcription factor 5 (ATF5) is a transcription factor that is highly expressed in undifferentiated neural progenitor/stem cells as well as a variety of human cancers including gliomas.AimsIn this study, we examined the expression and localization of ATF5 protein in canine gliomas, and targeting of ATF5 function in canine glioma cell lines.Materials and MethodsParaffin‐embedded canine brain glioma tissue sections and western blots of tumours and glioma cells were immunoassayed with anti‐ATF5 antibody. Viability of glioma cells was tested with a synthetic cell‐penetrating ATF5 peptide (CP‐d/n ATF5) ATF5 antagonist.Results
ATF5 protein expression was in the nucleus and cytoplasm and was present in normal adult brain and tumour samples, with significantly higher expression in tumours as shown by western immunoblotting. CP‐d/n ATF5 was found to decrease cell viability in canine glioma cell lines in vitro in a dose‐dependent manner.ConclusionSimilarities in expression of ATF5 in rodent, dog and human tumours, and cross species efficacy of the CP‐d/n ATF5 peptide support the development of this ATF5‐targeting approach as a novel and translational therapy in dog gliomas.
Biosynthesis of the Pharmaceutically Important Fungal Ergot Alkaloid Dihydrolysergic Acid Requires a Specialized Allele of
cloA
Authors: Arnold Stephanie L. , Panaccione Daniel G. 
Published: May 2017

ABSTRACT

Ergot alkaloids are specialized fungal metabolites that are important as the bases of several pharmaceuticals. Many ergot alkaloids are derivatives of lysergic acid (LA) and have vasoconstrictive activity, whereas several dihydrolysergic acid (DHLA) derivatives are vasorelaxant. The pathway to LA is established, with the P450 monooxygenase CloA playing a key role in oxidizing its substrate agroclavine to LA. We analyzed the activities of products of
cloA
alleles from different fungi relative to DHLA biosynthesis by expressing them in a mutant of the fungus
Neosartorya fumigata
that accumulates festuclavine, the precursor to DHLA. Transformants expressing CloA from
Epichloë typhina
×
Epichloë festucae
, which oxidizes agroclavine to LA, failed to oxidize festuclavine to DHLA. In substrate feeding experiments, these same transformants oxidized exogenously supplied agroclavine to LA, indicating that a functional CloA was produced. A genomic clone of
cloA
from
Claviceps africana
, a sorghum ergot fungus that produces a DHLA derivative, was cloned and expressed in the festuclavine-accumulating mutant of
N. fumigata
, but several introns in this genomic clone were not processed properly. Expression of a synthetic intron-free version of
C. africana
cloA
resulted in the accumulation of DHLA as assessed by fluorescence high-pressure liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS). In substrate feeding experiments, the
C. africana
CloA also accepted agroclavine as the substrate, oxidizing it to LA. The data indicate that a specialized allele of
cloA
is required for DHLA biosynthesis and that the pharmaceutically important compound DHLA can be produced in engineered
N. fumigata
.

IMPORTANCE
Ergot alkaloids are fungal metabolites that have impacted humankind historically as poisons and more recently as pharmaceuticals used to treat dementia, migraines, and other disorders. Much is known about the biosynthesis of ergot alkaloids that are derived from lysergic acid (LA), but important questions remain about a parallel pathway to ergot alkaloids derived from dihydrolysergic acid (DHLA). DHLA-derived alkaloids have minor structural differences compared to LA-derived alkaloids but can have very different activities. To understand how DHLA is made, we analyzed activities of a key enzyme in the DHLA pathway and found that it differed from its counterpart in the LA pathway. Our data indicate a critical difference between the two pathways and provide a strategy for producing DHLA by modifying a model fungus. The ability to produce DHLA in a model fungus may facilitate synthesis of DHLA-derived pharmaceuticals.
Moderate Expression of
SEC16
Increases Protein Secretion by Saccharomyces cerevisiae
Authors: Bao Jichen , Huang Mingtao , Petranovic Dina , Nielsen Jens 
Published: May 2017

ABSTRACT

The yeast
Saccharomyces cerevisiae
is widely used to produce biopharmaceutical proteins. However, the limited capacity of the secretory pathway may reduce its productivity. Here, we increased the secretion of a heterologous α-amylase, a model protein used for studying the protein secretory pathway in yeast, by moderately overexpressing
SEC16
, which is involved in protein translocation from the endoplasmic reticulum to the Golgi apparatus. The moderate overexpression of
SEC16
increased α-amylase secretion by generating more endoplasmic reticulum exit sites. The production of reactive oxygen species resulting from the heterologous α-amylase production was reduced. A genome-wide expression analysis indicated decreased endoplasmic reticulum stress in the strain that moderately overexpressed
SEC16
, which was consistent with a decreased volume of the endoplasmic reticulum. Additionally, fewer mitochondria were observed. Finally, the moderate overexpression of
SEC16
was shown to improve the secretion of two other recombinant proteins,
Trichoderma reesei
endoglucanase I and
Rhizopus oryzae
glucan-1,4-α-glucosidase, indicating that this mechanism is of general relevance.

IMPORTANCE
There is an increasing demand for recombinant proteins to be used as enzymes and pharmaceuticals. The yeast
Saccharomyces cerevisiae
is a cell factory that is widely used to produce recombinant proteins. Our study revealed that moderate overexpression of
SEC16
increased recombinant protein secretion in
S. cerevisiae
. This new strategy can be combined with other targets to engineer cell factories to efficiently produce protein in the future.
Epidemiology of competition injuries in youth karate athletes: a prospective cohort study
Authors: Čierna Dušana , Lystad Reidar P 
Published: May 2017

AimTo determine the injury incidence rate and injury pattern among youth karate athletes competing in national tournaments in Slovakia, and to identify risk factors for injury.MethodsData were collected at nine national youth karate tournaments in Slovakia in 2015 and 2016. Injury incidence rates were calculated per 1000 athlete-exposures (IIRAE) and per 1000 min of exposure (IIRME) with 95% CIs. Subgroups were compared by calculating their rate ratios (RR) with 95% CIs.ResultsThe overall IIRAEand IIRMEwere 45.3 (95% CI 38.7 to 52.6) and 35.9 (95% CI 30.7 to 41.7), respectively. The most frequently injured anatomical region was the head/neck (57%), while the most common type of injury was contusion (85%). The risk of injury for the 12–17-year-old age group was almost twice that of the 6–12-year-old age group, after accounting for exposure time (RRME1.92 (95% CI 1.39 to 2.65)), and the difference was more pronounced for girls than boys (RRME2.47 (95% CI 1.52 to 4.00) vs RRME1.62 (95% CI 1.06 to 2.49), respectively).ConclusionsYouth karate has a relatively large proportion of head injuries. Adolescent and female youth karate athletes are at higher risk of injury compared with their child and male counterparts.
Integrative Analysis Identifies a Novel AXL–PI3 Kinase–PD-L1 Signaling Axis Associated with Radiation Resistance in Head and Neck Cancer
Authors: Skinner Heath D. , Giri Uma , Yang Liang P. , Kumar Manish , Liu Ying , Story Michael D. , Pickering Curtis R. , Byers Lauren A. , Williams Michelle D. , Wang Jing , Shen Li , Yoo Suk Y. , Fan You Hong , Molkentine David P. , Beadle Beth M. , Meyn Raymond E. , Myers Jeffrey N. , Heymach John V. 
Published: May 2017

Abstract
Purpose: The primary cause of death due to head and neck squamous cell carcinoma (HNSCC) is local treatment failure. The goal of this study was to examine this phenomenon using an unbiased approach.
Experimental Design: We utilized human papilloma virus (HPV)-negative cell lines rendered radiation-resistant (RR) via repeated exposure to radiation, a panel of HPV-negative HNSCC cell lines and three cohorts of HPV-negative HNSCC tumors (n = 68, 97, and 114) from patients treated with radiotherapy and subjected to genomic, transcriptomic, and proteomic analysis.
Results: RR cell lines exhibited upregulation of several proteins compared with controls, including increased activation of Axl and PI3 kinase signaling as well as increased expression of PD-L1. Additionally, inhibition of either Axl or PI3 kinase led to decreased PD-L1 expression. When clinical samples were subjected to RPPA and mRNA expression analysis, PD-L1 was correlated with both Axl and PI3K signaling as well as dramatically associated with local failure following radiotherapy. This finding was confirmed examining a third cohort using immunohistochemistry. Indeed, tumors with high expression of PD-L1 had failure rates following radiotherapy of 60%, 70%, and 50% compared with 20%, 25%, and 20% in the PD-L1–low expression group (P = 0.01, 1.9 × 10−3, and 9 × 10−4, respectively). This finding remained significant on multivariate analysis in all groups. Additionally, patients with PD-L1 low/CD8+ tumor-infiltrating lymphocytes high had no local failure or death due to disease (P = 5 × 10−4 and P = 4 × 10−4, respectively).
Conclusions: Taken together, our data point to a targetable Axl–PI3 kinase–PD-L1 axis that is highly associated with radiation resistance. Clin Cancer Res; 23(11); 2713–22. ©2017 AACR.
Abstract A05: An in vivo model reveals a role for Argonaute 2 in oncogenic KRAS driven pancreatic cancer initiation
Authors: Shankar Sunita , Tien Jean , Dommeti Vijaya L. , Zelenka-Wang Sylvia , Chinnaiyan Arul M. 
Published: May 2017

Abstract
Oncogenic mutations in RAS provide a compelling yet intractable therapeutic target. Using co-immunoprecipitation mass spectrometry, we recently identified an interaction between RAS and Argonaute 2 (AGO2), the core component of the RNA silencing machinery exerting translational control of mRNA transcripts. RAS and AGO2 co-sediment and co-localize in intracellular endomembrane bound organelles. The AGO2 N-terminal domain directly binds the KRAS Switch II region, irrespective of GDP/GTP bound to RAS. Functionally, we observed that AGO2 was required for maximal oncogenic KRAS levels and AGO2 knock-down attenuates cell proliferation in mutant KRAS-dependent cells. Intriguingly, AGO2 mediated microRNA processing (RISC activity) was attenuated in cancer cells expressing mutant KRAS compared to those expressing wild type RAS. Yet, these initial investigations using cell line models do not reveal a clear role for AGO2 in KRAS driven oncogenesis. Therefore, we probed the role of AGO2 in the well-established pancreatic ductal adenocarcinoma (PDAC) mouse model driven by oncogenic KRAS.
Towards this end, we generated LSL-KrasG12D; Pft1a Cre; AGO2flox/flox (KCA) mice. Preliminary analysis (8-10 weeks of age) suggests that compared to LSL-KRASG12D (KC) mice, homozygous loss of AGO2 results in increased KRAS driven Acinar to Ductal Metaplasia (ADM) phenotype considered as precursors of PanIN (pancreatic intraepithelial neoplasia) and PDAC. This mirrors the phenotype of loss of Dicer, another microRNA processing enzyme, in the same model further reinforcing a role for microRNAs in restraining KRAS oncogenic programs. Yet unlike Dicer, loss of AGO2 alone did not cause any gross or histological changes in the development of the mouse pancreata. In an effort to understand the underlying mechanisms that are regulated by AGO2, we also studied the activated levels of a variety of signaling molecules in pancreata obtained from different genotypes. Using both Western blot and Immunohistochemical analyses, we demonstrate a previously unknown and critical role for AGO2 in KRAS-MAPK signaling pathway.
Together our data from the in vivo mouse model suggests that AGO2 is an important regulator of KRAS signaling and oncogenic KRAS alters the translational machinery through its interaction with AGO2.
Citation Format: Sunita Shankar, Jean Tien, Vijaya L. Dommeti, Sylvia Zelenka-Wang, Arul M. Chinnaiyan. An in vivo model reveals a role for Argonaute 2 in oncogenic KRAS driven pancreatic cancer initiation. [abstract]. In: Proceedings of the AACR Special Conference on Translational Control of Cancer: A New Frontier in Cancer Biology and Therapy; 2016 Oct 27-30; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2017;77(6 Suppl):Abstract nr A05.
Abstract A11: Profiling of miRNAs expression in pediatric brain tumors
Authors: Tantawy Marwa , Elzayat Mariam , Yehia Dina , Taha Hala 
Published: May 2017

Abstract
Introduction: The understanding of pediatric CNS tumors biology is essential in the development of disease stratification and in development of less toxic therapeutic agents as well as finding novel markers for early diagnosis. MicroRNAs are short 18-25 nucleotide small non-coding RNA molecules regulate gene expression. Recent studies showed that miRNAs play a significant role in brain tumor biology; and may up-regulated or down-regulated in malignancies, which referred their oncogenic or tumor suppressor effect. MiRNA expression patterns have been linked to clinical outcomes, tumor regulation; such as tumor progression, cell growth, cell death and metastasis. The identification of tumor- specific miRNA signatures may assist in future in the discovery of new biomarkers with diagnostic and prognostic utility. The main objective of the present study is to detect the expression of different miRNAs in different subtypes of pediatric CNS tumors to distinguish between them in discovering biomarkers for early detection in addition to develop novel therapies.
Methods: The expression level of 82 miRNAs were detected in 120 cases of pediatric CNS tumors from fixed formalin paraffin embedded tissues (FFPE), divided into four subtypes including; Low Grade Glioma, High Grade Glioma, Ependymoma, and Medulloblastoma using quantitative real time PCR (qRTPCR).
Results: Analysis of qRT-PCR data showed significant differences in miRNA expression between tumor subtypes with P value&lt; 0.05. Low expression of (miR-221, miR-9, and miR-181c/d) and over expression in miR-101, miR-222, miR-139, miR-1827 and miR-34c) in Medulloblastoma patients compared to other subtypes. Low expression of miR-10a and overexpression in (miR-10b, and miR-29a) in Ependymoma patients compared to other subtypes. Low expression of miR-26a and over expression in (miR-19a/b, miR-24, miR-27a, miR- 584 and miR-527) in Low Grade Glioma patients compared to other subtypes.
Conclusion: MicroRNAs are differentially expressed between different subtypes of pediatric CNS tumors suggesting that they may play a significant role in diagnosis to distinguish between different subtypes. A greater understanding of aberrant miRNA expression in pediatric brain tumors may support in the development of novel therapies. In addition, the characterization of tumor-specific miRNA signatures may play an important role in the discovery of biomarkers with diagnostic or prognostic utility.
Note: This abstract was not presented at the conference.
Citation Format: Marwa Tantawy, Mariam Elzayat, Dina Yehia, Hala Taha. Profiling of miRNAs expression in pediatric brain tumors. [abstract]. In: Proceedings of the AACR Special Conference on Translational Control of Cancer: A New Frontier in Cancer Biology and Therapy; 2016 Oct 27-30; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2017;77(6 Suppl):Abstract nr A11.