bluuebunny/crossref_metadata_2025 huggingface dataset:
Western Blotting with Solutions containing Nanoliter Volumes of Antibody
Authors: Cruz Walma David A. , Collins Joshua W.
Published: May 2019

Whether screening small mammal serum during antibody production or attempting to preserve a stock of precious antibody, this protocol's western blotting method using aliquots containing nanoliter volumes of antibody will benefit researchers. Time‐tested western blotting workflows allowing separation and analysis of proteins are routinely utilized in clinical and laboratory settings. The necessity for relatively large quantities of antibody is a major limitation to this universal tool. This article provides a step‐by‐step protocol for detecting proteins of interest with solutions containing nanoliter volumes of antibody without altering the preceding gel electrophoresis and transfer methods. Important considerations, frequently encountered problems, and means of optimizing reproducibility are discussed. Complementary diagrams, images, and videos are provided. The protocol is demonstrated using 0.3 nanoliters of anti‐serum to detect fibronectin in a human foreskin fibroblast cell line. Finally, two support protocols detailing methods of extracting proteins from cultured cells are reported. Published 2019. This article is a US Government work and is in the public domain in the USA.
Influence of Perfluorohexane‐Enriched Atmosphere on Viscoelasticity and Structural Order of Self‐Assembled Semifluorinated Alkanes at the Air‐Water Interface
Authors: Mielke Salomé , Abuillan Wasim , Veschgini Mariam , Liu Xianhe , Konovalov Oleg , Krafft Marie Pierre , Tanaka Motomu
Published: May 2019

Abstract: Semifluorinated alkanes FnHm self‐assemble into nanometer‐sized surface micelles at the air‐water interface. In this study, we investigated how an atmosphere enriched with perfluorohexane (PFH) influences the interfacial viscoelasticity and structural order of a monolayer of FnHm by the combination of dilational rheology and grazing‐incidence small‐angle X‐ray scattering (GISAXS). The monolayers behaved predominantly elastic which can be attributed to the strong dipole repulsions of the surface domains. Enrichment of the atmosphere with PFH lead to an increase of the compressibility and a decrease of the elastic modulus without altering the structural ordering of the FnHm molecules into highly correlated nanodomains, suggesting the adsorption of PFH molecules to the free spaces between the domains. The capability of FnHm domains to retain the structural integrity in the presence of PFH gas is promising for the fabrication of stable microbubbles for sonographic imaging.
Short‐Lived Radical Intermediates in the Photochemistry of Glucose Oxidase
Authors: Nag Lipsa , Lukacs Andras , Vos Marten H.
Published: May 2019

Glucose oxidase is a flavoprotein that is relatively well‐studied as a physico‐chemical model system. The flavin cofactor is surrounded by several aromatic acid residues that can act as direct and indirect electron donors to photoexcited flavin. Yet, the identity of the photochemical product states is not well established. We present a detailed full spectral reinvestigation of this issue using femtosecond fluorescence and absorption spectroscopy. Based on a recent characterization of the unstable tyrosine cation radical TyrOH•+, we now propose that the primary photoproduct involves this species, which was previously not considered. Formation of this product is followed by competing charge recombination and radical pair stabilization reactions that involve proton transfer and radical transfer to tryptophan. A minimal kinetic model is proposed, including a fraction of TyrOH.+ that is stabilized up to the tens of picoseconds timescale, suggesting a potential role of this species as intermediate in biochemical electron transfer reactions.
Return of the lysergamides. Part V: Analytical and behavioural characterization of 1‐butanoyl‐d‐lysergic acid diethylamide (1B‐LSD)
Authors: Brandt Simon D. , Kavanagh Pierce V. , Westphal Folker , Stratford Alexander , Elliott Simon P. , Dowling Geraldine , Wallach Jason , Halberstadt Adam L.
Published: May 2019

The psychedelic properties of lysergic acid diethylamide (LSD) have captured the imagination of researchers for many years and its rediscovery as an important research tool is evidenced by its clinical use within neuroscientific and therapeutic settings. At the same time, a number of novel LSD analogs have recently emerged as recreational drugs, which makes it necessary to study their analytical and pharmacological properties. One recent addition to this series of LSD analogs is 1‐butanoyl‐LSD (1B‐LSD), a constitutional isomer of 1‐propanoyl‐6‐ethyl‐6‐nor‐lysergic acid diethylamide (1P‐ETH‐LAD), another LSD analog that was described previously. This study presents a comprehensive analytical characterization of 1B‐LSD employing nuclear magnetic resonance spectroscopy (NMR), low‐ and high‐resolution mass spectrometry platforms, gas‐ and liquid chromatography (GC and LC), and GC‐condensed phase and attenuated total reflection infrared spectroscopy analyses. Analytical differentiation of 1B‐LSD from 1P‐ETH‐LAD was straightforward. LSD and other serotonergic hallucinogens induce the head‐twitch response (HTR) in rats and mice, which is believed to be mediated largely by 5‐HT2A receptor activation. HTR studies were conducted in C57BL/6J mice to assess whether 1B‐LSD has LSD‐like behavioral effects. 1B‐LSD produced a dose‐dependent increase in HTR counts, acting with ~14% (ED50 = 976.7 nmol/kg) of the potency of LSD (ED50 = 132.8 nmol/kg). This finding suggests that the behavioral effects of 1B‐LSD are reminiscent of LSD and other serotonergic hallucinogens. The possibility exists that 1B‐LSD serves as a pro‐drug for LSD. Further investigations are warranted to confirm whether 1B‐LSD produces LSD‐like psychoactive effects in humans.
Phosphodiesterase 4 Inhibition as a Therapeutic Target for Alcoholic Liver Disease: From Bedside to Bench
Authors: Rodriguez Walter E. , Wahlang Banrida , Wang Yali , Zhang Jingwen , Vadhanam Manicka V. , Joshi‐Barve Swati , Bauer Philip , Cannon Robert , Ahmadi Ali Reza , Sun Zhaoli , Cameron Andrew , Barve Shirish , Maldonado Claudio , McClain Craig , Gobejishvili Leila
Published: May 2019

Alcoholic liver disease (ALD) is a major cause of liver‐related mortality. There is still no US Food and Drug Administration–approved therapy for ALD, and therefore, identifying therapeutic targets is needed. Our previous work demonstrated that ethanol exposure leads to up‐regulation of cAMP‐degrading phosphodiesterase 4 (PDE4) expression, which compromises normal cAMP signaling in monocytes/macrophages and hepatocytes. This effect of ethanol on cAMP signaling contributes to dysregulated inflammatory response and altered lipid metabolism. It is unknown whether chronic alcohol consumption in humans alters hepatic PDE4 expression and cAMP signaling and whether inadequate cAMP signaling plays a pathogenic role in alcohol‐induced liver injury. Our present work shows that expression of the PDE4 subfamily of enzymes is significantly up‐regulated and cAMP levels are markedly decreased in hepatic tissues of patients with severe ALD. We also demonstrate the anti‐inflammatory efficacy of roflumilast, a clinically available PDE4 inhibitor, on endotoxin‐inducible proinflammatory cytokine production ex vivo in whole blood of patients with alcoholic hepatitis. Moreover, we demonstrate that ethanol‐mediated changes in hepatic PDE4 and cAMP levels play a causal role in liver injury in in vivo and in vitro models of ALD. This study employs a drug delivery system that specifically delivers the PDE4 inhibitor rolipram to the liver to avoid central nervous system side effects associated with this drug. Our results show that PDE4 inhibition significantly attenuates ethanol‐induced hepatic steatosis and injury through multiple mechanisms, including reduced oxidative and endoplasmic reticulum stress both in vivo and in vitro. Conclusion: Increased PDE4 plays a pathogenic role in the development of ALD; hence, directed interventions aimed at inhibiting PDE4 might be an effective treatment for ALD.
Vase‐Kite Equilibrium of Resorcin[4]arene Cavitands Investigated Using Molecular Dynamics Simulations with Ball‐and‐Stick Local Elevation Umbrella Sampling
Authors: Hahn David F. , Milić Jovana V. , Hünenberger Philippe H.
Published: May 2019

A key feature of resorcin[4]arene cavitands is their ability to switch between a closed/contracted (Vase) and an open/expanded (Kite) conformation. The mechanism and dynamics of this interconversion remains, however, elusive. In the present study, the Vase‐Kite transitions of a quinoxaline‐based and of a dinitrobenzene‐based resorcin[4]arene are investigated using molecular dynamics (MD) simulations in three environments (vacuum, chloroform, and toluene) and at three temperatures (198.15, 248.15, and 298.15 K). The challenge of sampling the Vase‐Kite transition, which occurs experimentally on the millisecond time scale, is overcome by calculating relative free energies using ball‐and stick local elevation umbrella sampling (B&S‐LEUS) to enhance the statistics on the relevant states and to promote interconversion transitions. Associated unbiased MD simulations also evidence for the first time a complete Vase‐to‐Kite transition, as well as transitions between degenerate Kite1 and Kite2 forms and solvent‐exchange events. The calculated Vase‐to‐Kite free‐energy changes ΔG are in qualitative agreement with the experimental magnitudes and trends. The level of quantitative agreement is, however, limited by the force‐field accuracy and, in particular, by the approximate treatment of intramolecular interactions at the classical level. The results are in line with a less stable Vase state for the dinitrobenzene compared to the quinoxaline compound, and a negative entropy change ΔS for the Vase‐to‐Kite transition of the latter compound. Relative free energies calculated for intermediates also suggest that the Vase‐Kite transition does not follow a concerted mechanism, but an asynchronous one with sequential opening of the flaps. In particular, the conformation involving two adjacent flaps open in a parallel direction (cis‐p) represents a likely intermediate, which has not been observed experimentally to date.
Breast cancer risk in BRCA1/2 mutation carriers and noncarriers under prospective intensified surveillance
Authors: Engel Christoph , Fischer Christine , Zachariae Silke , Bucksch Karolin , Rhiem Kerstin , Giesecke Jutta , Herold Natalie , Wappenschmidt Barbara , Hübbel Verena , Maringa Monika , Reichstein‐Gnielinski Simone , Hahnen Eric , Bartram Claus R. , Dikow Nicola , Schott Sarah , Speiser Dorothee , Horn Denise , Fallenberg Eva M. , Kiechle Marion , Quante Anne S. , Vesper Anne‐Sophie , Fehm Tanja , Mundhenke Christoph , Arnold Norbert , Leinert Elena , Just Walter , Siebers‐Renelt Ulrike , Weigel Stefanie , Gehrig Andrea , Wöckel Achim , Schlegelberger Brigitte , Pertschy Stefanie , Kast Karin , Wimberger Pauline , Briest Susanne , Loeffler Markus , Bick Ulrich , Schmutzler Rita K. ,   on behalf of the German Consortium for Hereditary Breast and Ovarian Cancer (GC‐HBOC)
Published: May 2019

Comparably little is known about breast cancer (BC) risks in women from families tested negative for BRCA1/2 mutations despite an indicative family history, as opposed to BRCA1/2 mutation carriers. We determined the age‐dependent risks of first and contralateral breast cancer (FBC, CBC) both in noncarriers and carriers of BRCA1/2 mutations, who participated in an intensified breast imaging surveillance program. The study was conducted between January 1, 2005, and September 30, 2017, at 12 university centers of the German Consortium for Hereditary Breast and Ovarian Cancer. Two cohorts were prospectively followed up for incident FBC (n = 4,380; 16,398 person‐years [PY], median baseline age: 39 years) and CBC (n = 2,993; 10,090 PY, median baseline age: 42 years). Cumulative FBC risk at age 60 was 61.8% (95% CI 52.8–70.9%) for BRCA1 mutation carriers, 43.2% (95% CI 32.1–56.3%) for BRCA2 mutation carriers and 15.7% (95% CI 11.9–20.4%) for noncarriers. FBC risks were significantly higher than in the general population, with incidence rate ratios of 23.9 (95% CI 18.9–29.8) for BRCA1 mutation carriers, 13.5 (95% CI 9.2–19.1) for BRCA2 mutation carriers and 4.9 (95% CI 3.8–6.3) for BRCA1/2 noncarriers. Cumulative CBC risk 10 years after FBC was 25.1% (95% CI 19.6–31.9%) for BRCA1 mutation carriers, 6.6% (95% CI 3.4–12.5%) for BRCA2 mutation carriers and 3.6% (95% CI 2.2–5.7%) for noncarriers. CBC risk in noncarriers was similar to women with unilateral BC from the general population. Further studies are needed to confirm whether less intensified surveillance is justified in women from BRCA1/2 negative families with elevated risk.
Label‐free bioanalysis of Leishmania infantum using refractive index tomography with partially coherent illumination
Authors: Soto Juan M. , Mas Alicia , Rodrigo José A. , Alieva Tatiana , Domínguez‐Bernal Gustavo
Published: May 2019

In this work, we report the use of refractive index (RI) tomography for quantitative analysis of unstained DH82 cell line infected with Leishmania infantum. The cell RI is reconstructed by using a modality of optical diffraction tomography technique that employs partially coherent illumination, thus enabling inherent compatibility with conventional wide‐field microscopes. The experimental results demonstrate that the cell dry mass concentration (DMC) obtained from the RI allows for reliable detection and quantitative characterization of the infection and its temporal evolution. The RI provides important insight for studying morphological changes, particularly membrane blebbing linked to an apoptosis (cell death) process induced by the disease. Moreover, the results evidence that infected DH82 cells exhibit a higher DMC than healthy samples. These findings open up promising perspectives for clinical diagnosis of Leishmania.
Retracted: Baicalein retards proliferation and collagen deposition by activating p38MAPK‐JNK via microRNA‐29
Authors: Yang Xiaoliang , Zhang Chunyan , Jiang Jinjie , Li Yinghao
Published: May 2019

Immoderate proliferation and deposition of collagen generally result in hypertrophic scars and even keloids. microRNA‐29 (miR‐29) has been proved as a crucial regulator in these pathological processes. Although mounting evidence have proved baicalein (BAI) impairs scar formation, it is still incompletely understood whether miR‐29 participated in the underlying mechanism. In the present study, NIH‐3T3 cells were stimulated with BAI, and then cell viability was analyzed by cell counting kit‐8 (CCK‐8) and Western blot. We further analyzed total soluble collagen, collagen 1, and alpha‐smooth muscle actin (α‐SMA) in NIH‐3T3 cells, which were exposed to transforming growth factor beta 1 (TGF‐β1)/BAI, using a Sircol assay kit, quantitative reverse transcription‐PCR (qRT‐PCR) and Western blot, respectively. Besides, the miR‐29 inhibitor was transduced and its transfection efficiency was verified by qRT‐PCR. Finally, the phosphorylated p38 mitogen‐activated protein kinase (p38MAPK) and c‐Jun N‐terminal kinase (JNK) were examined by Western blot. BAI effectively retarded NIH‐3T3 proliferation in a dose‐dependent manner. Besides, TGF‐β1‐induced deposition of total soluble collagen and synthesis of collagen 1 and α‐SMA were repressed by BAI at mRNA and protein levels. However, miR‐29 inhibitor reversed the effects of BAI. Remarkably, BAI promoted phosphorylated expression of p38MAPK and JNK while miR‐29 inhibitor reversed its effects on the phosphorylated expression of p38MAPK and JNK. BAI effectively weakened the cell viability and repressed TGF‐β1‐induced total soluble collagen as well as collagen 1 and α‐SMA by upregulating miR‐29. Mechanically, BAI activates the p38MAPK/JNK pathway by promoting miR‐29.
Trifluoperazine as an alternative strategy for the inhibition of tumor growth of colorectal cancer
Authors: Qian Kun , Sun Laiyu , Zhou Guoqing , Ge Haixia , Meng Yue , Li Jingfen , Li Xiao , Fang Xinqiang
Published: May 2019

The development of cancer in patients with schizophrenia is affected by genetic and environmental factors and antipsychotic medication. Several studies found that schizophrenia was associated with decreased risk of some cancers, and the neuroleptic medication might help to reduce the risk of colorectal cancer (CRC). Phenothiazine drugs including trifluoperazine (TFP) are widely used antipsychotic drugs and showed some antitumor effects, we here investigated the potential application of TFP in the treatment of colon cancer. A series doses of TFP were treated to the colon cancer cell line HCT116 and the inhibitory concentration (IC50) of TFP for HCT116 was determined by cell counting kit‐8. The results indicated that the treatment of TFP impaired the cell vitality of HCT116 in a dose‐ and time‐dependent manner. Meanwhile, the Edu assay demonstrated that the proliferation was also inhibited by TFP, which was accompanied with the induction of apoptosis and autophagy. The expression of CCNE1, CDK4, and antiapoptosis factor BCL‐2 was downregulated but the proapoptosis factor BAX was upregulated. The autophagy inhibitor chloroquine could significantly reverse the TFP‐induced apoptosis. Moreover, the ability of migration and invasion of HCT116 was found to be suppressed by TFP, which was associated with the inhibition of epithelial‐mesenchymal transition (EMT). The function of TFP in vivo was further confirmed. The results showed that the administration of TFP remarkably abrogated the tumor growth with decreased tumor volume and proliferation index Ki‐67 level in tumor tissues. The EMT phenotype was also confirmed to be inhibited by TFP in vivo, suggesting the promising antitumor effects of TFP in CRC.
Peimine inhibits the growth and motility of prostate cancer cells and induces apoptosis by disruption of intracellular calcium homeostasis through Ca2+/CaMKII/JNK pathway
Authors: Tan Hailin , Zhang Guiming , Yang Xuecheng , Jing Tao , Shen Daqing , Wang Xinsheng
Published: May 2019

Prostate cancer (PC) is one of the most common malignant tumors in man. Peimine (PM) is a bioactive substance isolated from Fritillaria. Previous studies have shown that PM could inhibit the occurrence of a variety of cancers. However, the roles of PM in PC and its related mechanism have not been elucidated. Calcium (Ca2+) is an important intracellular messenger involved in a variety of cell processes. In this study, we found that the appropriate doses of PM (2.5, 5, and 10 μM) significantly inhibited the growth of PC cells (DU‐145, LNCap, and PC‐3), but has no significant effect on normal prostate cells (RWPE‐1). In addition, PM treatment inhibited the invasion and migration of PC‐3 cells and blocked the epithelial‐mesenchymal transition process. These effects were exhibited a dose‐dependent manner. Furthermore, the current results also showed that PM treatment significantly increased the Ca2+ concentration, the increased Ca2+ promoted the phosphorylation of Ca2+/calmodulin‐dependent protein kinase II (CaMKII) and c‐Jun N‐terminal kinase (JNK), further inhibited the growth and invasion of PC‐3 cells, and induced its apoptosis. Ca2+ chelator BAPTA‐AM (1 μM) could counteract the increase of intracellular Ca2+ concentration. Similarly, JNK pathway inhibitor SP600125 (10 μM) also inhibited cell growth and invasion and induced apoptosis. In addition, experiments in nude mice showed that PM inhibited tumor formation through Ca2+/CaMKII/JNK signaling pathway. In conclusion, our results show that PM inhibits the growth and motility of prostate cancer cells and induces apoptosis by disruption of intracellular calcium homeostasis through Ca2+/CaMKII/JNK pathway.
An ego network analysis approach identified important biomarkers with an association to progression and metastasis of gastric cancer
Authors: Tian Xiaofeng , Ju Haiying , Yang Wei
Published: May 2019

Background

Gastric cancer (GC) is the fifth most common cancer type worldwide. The aim of this study was to identify gastric‐related therapeutic indicators on the basis of the ego network analysis.

Material and methods

The microarray data related to GC was downloaded from ArrayExpress database. All human protein‐protein interaction (PPI) networks were downloaded from the STRING database. Ego genes were identified on the basis of PPI networks and the gene expression in GC, and then co‐expression networks (ego networks) were constructed using these ego genes. On the basis of ego networks, the optimal GO terms and genes were predicted by affinity predictions and cold read predictions. Finally, the predicted genes as effective biomarkers for GC were verified by the bioinformatics analysis.

Results

The differential expression networks were conducted and comprised of 365 edges and 232 nodes, which resulted in 218 ego genes. Although there was no significant difference in the expression of top ten ego genes among different groups of GC samples, it was eventually confirmed that top three optimal GO terms with highest cool read values were translational termination (cool read value = 0.987), translational elongation (cool read value = 0.986), and macromolecular complex disassembly (cool read value = 0.985) and top five optimal genes were UBA52, RPS27A, MAPK1, UBC, and UBB. UBA52, RPS27A, and MAPK1 were verified by the bioinformatics analysis to be related to the progression and metastasis of GC.

Conclusions

An ego network analysis approach is a very effective method for screening GC and the screened genes might be biomarkers for GC diagnosis and treatment.
Research on the roles of genes coding ATP‐binding cassette transporters in Porphyromonas gingivalis pathogenicity
Authors: Gao Li , Ma Yuanyuan , Li Xiting , Zhang Liping , Zhang Chi , Chen Qianying , Zhao Chuanjiang
Published: May 2019

Porphyromonas gingivalis, as a major pathogen of periodontitis, could rapidly adhere to and invade host gingival epithelial cells (GECs) for the induction of infection. One ATP‐binding cassette (ABC) transporter gene was found to be upregulated during this infection process, however, the molecular mechanisms remain unclear. In this study, we systemically investigated the messenger RNA level changes of all ABC transporter family genes in P. gingivalis while being internalized within GECs by real‐time polymerase chain reaction. We identified that two ABC transporter genes, PG_RS04465 (PG1010) and PG_RS07320 (PG1665), were significantly increased in P. gingivalis after coculturing with GECs. Mutant strains with knockout (KO) of these two genes were generated by homogenous recombination. PG_RS04465 and PG_RS07320 KO mutants showed no change in the growth of bacteria per se. Knockdown of PG_RS07320, but not PG_RS04465, caused decreased endotoxin level in the bacteria. In contrast, both mutant strains showed decreased Arg‐ and Lys‐gingipains activities, with significantly reduced adhesion and invasion capabilities. Secreted interleukin‐1β (IL‐1β) and IL‐6 levels in GECs cocultured with PG_RS04465 or PG_RS07320 KO mutants were also decreased, whereas, only the cells cocultured with PG_RS07320 KO mutants showed significant decrease. In addition, virulence study using mouse revealed that both KO mutant strains infection caused less mouse death than wild‐type strains, showing reduced virulence of two KO strains. These results indicated that ABC transporter genes PG_RS04465 and PG_RS07320 are positive regulators of the virulence of P. gingivalis.
MicroRNAs and response to therapy in leukemia
Authors: Sharifi Hossein , Jafari Najaf Abadi Mohammad Hassan , Razi Ebrahim , Mousavi Nousin , Morovati Hamid , Sarvizadeh Mostafa , Taghizadeh Mohsen
Published: May 2019

A variety of epigenetic factors involved in leukemia pathogenesis. Among various epigenetic factors, microRNAs (miRNAs) have emerged as important players, which affect a sequence of cellular and molecular signaling pathways. Leukemia is known as progressive cancer, which is related to many health problems in the world. It has been shown that the destruction of the blood‐forming organs could lead to abnormal effects on the proliferation and development of leukocytes and their precursors. Despite many attempts for approved effective and powerful therapies for patients with leukemia, finding and developing new therapeutic approaches are required. One of the important aspects of leukemia therapy, identification of underlying cellular and molecular mechanisms involved in the pathogenesis of leukemia. Several miRNAs (ie, miR‐103, miR‐101, mit‐7, let‐7i, miR‐424, miR‐27a, and miR‐29c) and play major roles in response to therapy in patients with leukemia. miRNAs exert their effects by targeting a variety of targets, which are associated with response to therapy in patients with leukemia. It seems that more understanding about the roles of miRNAs in response to therapy in patients with leukemia could contribute to better treatment of patients with leukemia. Here, for the first time, we summarized various miRNAs, which are involved in response to therapy in the treatment patients with leukemia.
Comparative assessment of the therapeutic drug targets of C. botulinum ATCC 3502 and C. difficile str. 630 using in silico subtractive proteomics approach
Authors: Bhardwaj Tulika , Haque Shafiul , Somvanshi Pallavi
Published: May 2019

Growing antimicrobial resistance of the pathogens against multiple drugs posed a serious threat to the human health worldwide. This fueled the need of identifying the novel therapeutic targets that can be used for developing new class of the drugs. Recently, there is a substantial rise in the rate of Clostridium infections as well as in the emergence of virulent and antibiotic resistant strains. Hence, there is an urgent need for the identification of potential therapeutic targets and the development of new drugs for the treatment and prevention of Clostridium infections. In the present study, a combinatorial approach involving systems biology and comparative genomics strategy was tested against Clostridium botulinum ATCC 3502 and Clostridium difficile str. 630 pathogens, to render potential therapeutic target at qualitative and quantitative level. This resulted in the identification of five common (present in both the pathogens, 34 in C. botulinum ATCC 3502 and 42 in C. difficile str. 630) drug targets followed by virtual screening–based identification of potential inhibitors employing molecular docking and molecular dynamics simulations. The identified targets will provide a solid platform for the designing of novel wide‐spectrum lead compounds capable of inhibiting their catalytic activities against multidrug‐resistant Clostridium in the near future.
TLR4 and nucleolin influence cell injury, apoptosis and inflammatory factor expression in respiratory syncytial virus‐infected N2a neuronal cells
Authors: Hu Tao , Yu Haiyang , Lu Min , Yuan Xiaoling , Wu Xuan , Qiu Huan , Chen Jason , Huang Shenghai
Published: May 2019

Respiratory syncytial virus (RSV) infection was recently reported to be associated with central nervous system (CNS) symptoms and neurological complications; however, related studies are very limited. Moreover, the molecular mechanism underlying RSV neuropathogenesis is still unclear. Our previous study revealed that toll‐like receptor 4 (TLR4) and nucleolin (C23) could be modulated and that they played a role during RSV infection in mouse neuronal‐2a (N2a) cells. In the present study, the effects of silencing of TLR4 and C23 on RSV propagation and N2a cellular responses were examined by using RNA interference technology. Four N2a cell treatment groups were established, namely, a normal control group, RSV control group, TLR4 siRNA + RSV group, and C23 siRNA + RSV group. Expression changes in NeuN protein and colocalization of C23 and TLR4 with RSV F protein were assessed using confocal microscopy. Changes in TLR4 and C23 mRNA expression, TLR4, C23, TLR3, TLR7, and p‐NF‐κB protein expression, and interleukin (IL)‐8, IL‐6, and tumor necrosis factor (TNF‐α) cytokine secretion was measured using quantitative real‐time reverse‐transcription polymerase chain reaction, Western blot analysis, and enzyme‐linked immunosorbent assay, respectively. RSV titers and the apoptotic status of N2a cells were monitored using plaque formation assays and flow cytometry, respectively. The results indicated that TLR4 and C23 gene knockdown decreased the amount of F protein in RSV‐infected N2a cells, inhibited RSV propagation, attenuated N2a neuronal injury, diminished cell apoptosis levels, downregulated TLR3 and TLR7 protein expression, and reduced inflammatory protein expression. Therefore, TLR4 and C23 knockdown influences cell injury, apoptosis and inflammatory protein expression in RSV‐infected N2a cells.
Retracted: Elevation of circular RNA circ‐POSTN facilitates cell growth and invasion by sponging miR‐1205 in glioma
Authors: Yang Yimin , Zhang Ying , Chen Bin , Ding Lili , Mu Zhuang , Li Yanhua
Published: May 2019

Glioma is the most common type of primary intracranial tumor. Dysregulation of circular RNAs (circRNAs) plays a critical role in multiple solid tumors. However, the expression profiles of circRNAs and their functions in glioma have been rarely studied. The current work aims to investigate the clinical significance of a novel circRNA, circ‐POSTN, in glioma and explore its biological functions and mechanisms in the progression of glioma. We found that circ‐POSTN was highly expressed in glioma tissue samples and cells. High circ‐POSTN expression was significantly linked to larger tumor size, higher World Health Organization grades, and shorter overall survival. Furthermore, silencing of circ‐POSTN in glioma cells could decrease cell growth, migratory and invasive potential, and induce cell apoptosis in LN229 cells. On the contrary, ectopically expressed circ‐POSTN induced the opposite effects in the U251 cell line. By bioinformatic prediction and luciferase reporter assay, we identified that miR‐1205 could be sponged by circ‐POSTN. Further rescue assays demonstrated that the oncogenic functions of circ‐POSTN are partly attributed to its regulation of miR‐1205 in glioma cells. Taken together, our data suggest that circ‐POSTN plays an oncogenic role in glioma progression and may serve as a novel therapeutic target in this deadly disease.
Identification of an adaptor protein‐2 mu gene (AccAP2m) in Apis cerana cerana and its role in oxidative stress responses
Authors: Wang Xinxin , Chao Yuzhen , Wang Ying , Xu Baohua , Wang Chen , Li Han
Published: May 2019

The adaptor protein‐2 (AP‐2) complex refers to a class of proteins that play an important role in cargo recognition and are involved in clathrin‐mediated endocytosis of cargo proteins from the plasma membrane in animal cells. However, there have been few studies of AP‐2 in insects. Here, we isolated an AP‐2 gene from Apis cerana cerana (A. cerana cerana) and explored the function of this gene, named AccAP2m. The obtained amino acid sequence of AccAP2m is highly conserved across species and contains conserved features of the AP‐2 family. Quantitative real‐time PCR analysis showed that AccAP2m mRNA levels were high in newly emerged workers, and the expression levels were higher in muscle than in other tissues. The expression of AccAP2m was altered in response to various environmental stress factors. Moreover, overexpression of recombinant AccAP2m protein enhanced the resistance of the bacteria to HgCl2, CdCl2, and cumyl hydroperoxide. In addition, after knockdown of AccAP2m in A. cerana cerana by RNA interference (RNAi), the transcription of some antioxidant genes was downregulated, and the activities of some antioxidant enzymes decreased. Taken together, these results indicate that AccAP2m may play an indispensable role in response to oxidative stresses.
Tid1‐S attenuates LPS‐induced cardiac hypertrophy and apoptosis through ER‐a mediated modulation of p‐PI3K/p‐Akt signaling cascade
Authors: Chao Chun‐Nun , Lo Jeng‐Fan , Khan Farheen B. , Day Cecilia H. , Lai Chao‐Hung , Chen Chia‐Hua , Chen Ray‐Jade , Viswanadha Vijaya P. , Kuo Chia‐Hua , Huang Chih‐Yang
Published: May 2019

Myocardial dysfunction is clinically relevant? repercussion that follows sepsis. Tid 1 protein has been implicated in many biological process. However, the role of Tid 1 in lipopolysaccharide (LPS)‐induced cardiomyocyte hypertrophy and apoptosis remains elusive. In the current research endeavor, we have elucidated the role of Tid1‐S on LPS‐induced cardiac hypertrophy and apoptosis. Interestingly, we found that overexpression of Tid1‐S suppressed TLR‐4, NFATc3, and BNP protein expression which eventually led to inhibition of LPS‐induced cardiac hypertrophy. Moreover, Tid1‐S overexpression attenuated cellular apoptosis and activated survival proteins p‐PI3K and pser473Akt. Besides this, Tid1‐S overexpression enhanced ER‐a protein expression. Collectively, our data suggest that Tid1‐S plausibly enhance ER‐a protein and further activate p‐PI3K and p
ser473Akt survival protein expression; which thereby led to attenuation of LPS‐induced apoptosis in cardiomyoblast cells. Interestingly, our data suggest that Tid1‐S is involved in attenuation of cardiomyoblast cells damages induced by LPS.
Effect of embryo cryopreservation on derivation efficiency, pluripotency, and differentiation capacity of mouse embryonic stem cells
Authors: Assadollahi Vahideh , Hassanzadeh Kambiz , Abdi Mohammad , Alasvand Masoud , Nasseri Sherko , Fathi Fardin
Published: May 2019

Mouse embryonic stem cells (mESCs) are pluripotent cells that have the capability for self‐renewal. One of the most important factors that affect the efficiency of their isolation is the condition of the mouse embryos. The main objective of this study is to isolate mESCs from C57BL/6 frozen/thawed eight‐cell mouse embryos using serum‐free culture. We generated mESCs from blastocysts that developed from frozen/thawed embryos of C57BL/6 mice by the 3i + LIF medium. Assessments of the isolated mESC lines (MUKF‐1, MUKF‐2, and MUKF‐3) included simple karyotype analysis; polymerase chain reaction of the testis‐determining gene (Sry); determination of alkaline phosphatase (ALP) activity; expressions of pluripotent transcription factors Oct4, Rex1, Sox2, and Nanog by reverse transcription polymerase chain reaction; and immunocytochemistry assessment of OCT4 and SSEA‐I expressions at the protein level. We evaluated the ability of these mESC lines to differentiate into three germ layers by embryoid body (EB) formation. The cell doubling time (DT) of isolated mESCs was determined. The 2‐C57 cell line was served as control. Germline competence of the male mESC line (MUKF‐3) was tested through chimeric mouse production. Three independent mESC lines (MUKF‐1, MUKF‐2, and MUKF‐3) were established from five cryopreserved embryos. The MUKF‐1 and MUKF‐2 lines were female, whereas MUKF‐3 was a male mESC line. Karyotype analysis showed that MUKF‐3 had a diploid karyotype, whereas MUKF‐1 and MUKF‐2 had abnormal karyotypes. All three lines had ALP activity and expressed Oct4, Rex1, and Nanog. Immunocytochemistry assessment for OCT4 and SSEA‐I was positive for all three lines. The DT differed in the three mESC lines. MUKF‐1 and MUKF‐3 could form EB and express developmental genes after spontaneous differentiation. These data demonstrated that probably cryopreservation affected the efficiency of derivation, karyotype, DT, expression of pluripotency, developmental genes, and differentiation capacity of the independent mESC lines.
Integrative analyses of triple negative dysregulated transcripts compared with non‐triple negative tumors and their functional and molecular interactions
Authors: Darbeheshti Farzaneh , Rezaei Nima , Amoli Mahsa M. , Mansoori Yaser , Tavakkoly Bazzaz Javad
Published: May 2019

Triple‐negative (TN) tumors are a subtype of breast cancer with aggressive behaviors and limited targeted therapies. Microarray studies were not concerned with interactions and functional relations of dysregulated transcripts. Here, we aimed to conduct integrative strategy to analyze gene and miRNA available microarray data as well as bioinformatic analyses to catch a more inclusive picture of pivotal dysregulated transcripts and their interactions in TN tumors. Several online datasets and offline bioinformatic tools were used to detect differentially expressed (DE) transcripts, both protein and nonprotein coding, in TN compared with non‐TN tumors and their functional and molecular interactions.

Sixteen upregulated and 58 downregulated genes with a log fold change higher or equal to | 2 | were identified, including nine transcription factors. Coexpression network revealed EN1 as a hub gene, moreover Kaplan–Meier plotter survival analysis indicated that it was an appropriate prognostic marker for TN patients with breast cancer. Functional annotation analysis of protein–protein interaction network showed FOXM1 as an upexpressed and ESR1 as a downexpressed hub genes are suitable targets as far as antitumor protein therapy is concerned in TN breast cancers. The consensus analysis of two microRNA datasets revealed seven DE miRNAs. The gene–transcriptional factor (TF)–miRNA network revealed mir‐135b and mir‐29b are the hub nodes and involved in feedback loops with GATA3. This study suggests that dysregulated TFs and miRNAs have pivotal roles in regulation of TN oncotranscriptomic profile and might become both biomarkers and therapeutic targets.
Association between TL1A gene polymorphisms and systemic lupus erythematosus in a Chinese Han population
Authors: Xu Wang‐Dong , Fu Lu , Liu Xiao‐Yan , Wang Jia‐Min , Yuan Zhi‐Chao , Su Lin‐Chong , Huang An‐Fang
Published: May 2019

Our previous studies showed elevated tumor necrosis factor‐like ligand 1 aberrance (TL1A) expression in systemic lupus erythematosus (SLE). However, TL1A polymorphisms with SLE susceptibility remain to be elucidated. In addition, we made meta‐analysis to evaluate the relationship of TL1A polymorphisms and autoimmune diseases owing to inconsistent results. The present research was carried out by 404 SLE, 150 primary Sjogren's syndrome (pSS) patients, and 574 healthy individuals. Three TL1A polymorphisms (rs3810936, rs6478109, rs7848647) were genotyped using TaqMan genotyping assay. Then, the meta‐analysis was performed by collecting the present case‐control study and previously published research. Results showed that genotypes of rs3810936, rs7848647 were different between SLE patients and healthy controls, whereas no significant association was observed in the three polymorphisms and pSS patients. Genotypes distribution of rs6478109, rs7848647 were strongly related to lupus nephritis within SLE (p = 0.004, p = 0.011), respectively. Moreover, combined meta‐analysis consisted of ten comparative research involving 4,305 patients and 5,600 controls. An association between autoimmune diseases and rs6478109 polymorphism was found. Our findings indicate that gene polymorphisms (rs3810936, rs7848647) of TL1A might correlate with lupus.
The spectrum of viral pathogens in children with severe acute lower respiratory tract infection: A 3‐year prospective study in the pediatric intensive care unit
Authors: Li Ya‐Ting , Liang Ying , Ling Ye‐Sheng , Duan Meng‐Qi , Pan Li , Chen Zhuang‐Gui
Published: May 2019

Background

No comprehensive analysis is available on the viral etiology and clinical characterization among children with severe acute lower respiratory tract infection (SALRTI) in Southern China.

Methods

Cohort of 659 hospitalized children (2 months to 14 years) with SALRTI admitted to the Pediatric Intensive Care Unit (PICU) in the Guangzhou from May 2015 to April 2018 was enrolled in this study. Nasopharyngeal aspirate specimens or induced sputum were tested for eight categories respiratory viral targets. The viral distribution and its clinical characters were statistically analyzed.

Results

Viral pathogen was detected in 326 (49.5%) of children with SALRTI and there were 36 (5.5%) viral coinfections. Overall, the groups of viruses identified were, in descending order of prevalence: Influenza virus (IFV) (n = 94, 14.3%), respiratory syncytial virus (RSV) (n = 75, 11.4%), human rhinovirus (HRV) (n = 56, 8.5%), adenovirus (ADV) (n = 55, 8.3%), parainfluenza (PIV) (n = 47, 7.1%), human coronavirus (HCoV) (n = 15, 2.3%), human metapneumovirus (HMPV) (n = 14, 2.1%) and human bocavirus (HBoV) (n = 11, 1.7%). The positive rate in younger children (< 5 years) was significantly higher than the positive rate detected in elder children (> 5 years) (52.5% vs 35.1%, P = 0.001). There were clear seasonal peaks for IFV, RSV, HRV, ADV, PIV, and HMPV. And the individuals with different viral infection varied significantly in terms of clinical profiles.

Conclusions

Viral infections are present in a consistent proportion of patients admitted to the PICU. IFV, RSV, HRV, and ADV accounted for more than two‐thirds of all viral SALRTI. Our findings could help the prediction, prevention and potential therapeutic approaches of SALRTI in children.
The characteristics of moisture recycling and its impact on regional precipitation against the background of climate warming over Northwest China
Authors: Wu Ping , Ding Yihui , Liu Yanju , Li Xiucang
Published: May 2019

From 1961 to 2015, annual and seasonal precipitation have shown a significant increase over Northwest China (NW). The change of precipitation is a combination result of the changing recycling precipitation (Pw) contributed by local evapotranspiration (ET) and changing precipitation contributed by advection moisture (Po). In this paper, we calculated the ET based on the observation data of meteorological stations and the complementary relationship model. Then used an improved precipitation recycling model to analyse influential factors on the variation of precipitation over the NW from aspects of ET and advection moisture. The annual precipitation recycling ratio (ρ) fluctuates between 4% and 10% and shows an increasing trend at a rate of 0.3% /10a with a significance level of 0.01. ET, which is the internal supplier of the precipitable water, and advection moisture, which is the outside supplier of the precipitable water, both have displayed significantly increasing trends over the NW. The advection moisture contributes precipitation much more than ET, no matter in the annual scale or seasonal scales. In view of the spatial distribution, Pw shows an increasing trend over the whole NW, but distinctly opposite spatial patterns of Po are found between the western area of NW (WNW) and the eastern area of NW (ENW). Such differences caused the precipitation is characterized by a wetting trend in WNW but a drying trend in ENW.
Ovulatory follicular fluid induces sperm phagocytosis by neutrophils, but oviductal fluid around oestrus suppresses its inflammatory effect in the buffalo oviduct in vitro
Authors: Yousef Mohamed S. , Abd‐Elhafeez Hanan H. , Talukder Anup K. , Miyamoto Akio
Published: May 2019

We have recently shown that the conditioned media from bovine oviductal epithelial cell culture suppress sperm phagocytosis by neutrophils, suggesting that the oviduct around oestrus supplies the anti‐inflammatory microenvironment. To investigate the immune response of neutrophils toward the sperm at ovulation in the buffalo oviduct, we examined (a) a detailed distribution of neutrophils in the oviduct in buffaloes, (b) the effect of ovulatory follicular fluid (FF) and oviductal fluid (OF) on sperm phagocytosis by neutrophils, and (c) the interaction of the ovulatory FF with OF on sperm phagocytosis by neutrophils in vitro. Buffalo oviducts were collected from healthy reproductive tracts at a local slaughterhouse. A detailed observation by histological examination and transmission electron microscopy revealed that neutrophils exist in the oviduct epithelium and lumen throughout the oestrous cycle in buffaloes. The number of neutrophils at the oestrus stage was higher in ampulla compared with those in isthmus, whereas they remained relatively constant at the dioestrus stage. Two hours of preincubation of neutrophils with FF enhanced sperm phagocytosis through the formation of neutrophil extracellular traps (NETs) together with H2O2 production, whereas OF around oestrus (eOF) suppressed sperm phagocytosis, NETs formation, and H2O2 production and relieved the above FF‐induced inflammatory response. Our findings show that neutrophils exist in the healthy cyclic oviduct across bovine species, and the OF supplies a strong anti‐inflammatory environment that could minimize the inflammatory effect of the FF that flows into the oviduct lumen after ovulation and supports the occurrence of fertilization.
Specialized naphthoquinones present in Impatiens glandulifera nectaries inhibit the growth of fungal nectar microbes
Authors: Block Anna K. , Yakubova Elena , Widhalm Joshua R.
Published: May 2019

The invasion success of Impatiens glandulifera (Himalayan balsam) in certain parts of Europe and North America has been partially attributed to its ability to compete for bee pollinators with its rich nectar and due to its capacity to produce and release allelopathic 1,4‐naphthoquinones (1,4‐NQs) from its roots and leaves. Given that other 1,4‐NQs present in the digestive fluids of certain carnivorous plants are proposed to control microbial colonization, we investigated the potential for the 1,4‐NQs, 2‐methoxy‐1,4‐naphthoquinone (2‐MNQ) and lawsone, to fulfill an analogous role in the nectaries of I. glandulifera. Both 2‐MNQ and lawsone were detected in the floral nectaries of I. glandulifera at levels comparable to leaves and roots, but were discovered to be at significantly higher levels in its extra‐floral nectaries (EFNs) and to be present in EFN nectar itself. Nectar microbe inhibition assays revealed that the common nectar bacteria Gluconobacter oxydans and Asaia prunellae are not inhibited by 2‐MNQ or lawsone, although both compounds were found to inhibit the growth of the common fungal nectar microbes Metschnikowia reukaufii and Aureobasidium pullulans. Taken together, these findings suggest that 2‐MNQ and lawsone could serve to protect the rich nectar of I. glandulifera against fungal growth. The high abundance of 2‐MNQ and lawsone in I. glandulifera EFNs may also point to an unsuspected mechanism for how allelopathic 1,4‐NQs are leached into the soil where they exhibit their known allelopathic effects.
Genetic basis of surfactant dysfunction in Chinese children: A retrospective study
Authors: Chen Jiehua , Nong Guangmin , Liu Xiuyun , Ji Wei , Zhao Deyu , Ma Hongling , Wang Heping , Zheng Yuejie , Shen Kunling
Published: May 2019

Objective

To investigate the prevalence of surfactant dysfunction (SD) and the genotype distribution in Chinese childhood interstitial lung disease (chILD).

Methods

From December 2013 to December 2016, whole exons and splicing regions of surfactant protein (SP)‐B, SP‐C, and adenosine triphosphate (ATP)‐binding cassette subfamily A member 3 (ABCA3) were sequenced in chILD with unknown etiology in five children's medical centers of China. The sequencing was performed by Next‐generation sequencing technique in a molecular genetics laboratory. The clinical and genetic data were reviewed retrospectively.

Results

In total, 136 patients of age 3 months to 13 years (mean 12.5 ± 9.4 months) were recruited, among which 76 were males. Of the 136 cases of chILD, 13.2% (18 of 136) were diagnosed with SD. In these 18 SD cases, 15 had heterozygous SP‐C deficiencies, two cases had compound heterozygous ABCA3 deficiencies, and no SP‐B deficiency was identified. In SP‐C deficiencies, there were six cases with p.I73T, 2 with p.I73N, 5 with p.V39L, 1 with c.417delA, and 1 case with IVS4, +1G>C. Two cases of ABCA3 mutation were heterozygous with c.1755delC and c.2890G>A; c.3913T>C (R1305W) and exon 13 to 18 deletion. One was negative by sequencing while diagnosed positive by pathology.

Conclusion

The proportion of genetic mutation of SD in chILD is 13.2% in China, of which SP‐C deficiency is predominant. The mutation, SP‐C p.V39L, was found to be relatively prevalent in China and warrants further investigation.