bluuebunny/crossref_metadata_2025 huggingface dataset:
Chromium hazard and risk assessment: New insights from a detailed speciation study in a standard test medium
Authors: Aharchaou Imad , Py Jean-Sébastien , Cambier Sébastien , Loizeau Jean-Luc , Cornelis Geert , Rousselle Philippe , Battaglia Eric , Vignati Davide A.L.
Published: November 2017

Despite the consensus about the importance of chemical speciation in controlling the bioavailability and ecotoxicity of trace elements, detailed speciation studies during laboratory ecotoxicity testing remain scarce, contributing to uncertainty when extrapolating laboratory findings to real field situations in risk assessment. We characterized the speciation and ecotoxicological effects of chromium (CrIII and CrVI) in the International Organization for Standardization (ISO) medium for algal ecotoxicity testing. Total and dissolved (< 0.22 μm) Cr concentrations showed little variability in media spiked with CrVI, whereas dissolved Cr concentration decreased by as much as 80% over a 72-h time period in medium amended with CrIII. Analyses by ion chromatography inductively coupled plasma mass spectrometry (IC-ICP-MS) highlighted the absence of redox interconversion between CrIII or CrVI both in the presence and absence of algal cells (Raphidocelis subcapitata). Furthermore, the concentration of ionic CrIII dropped below detection limits in less than 2 h with the corresponding formation of carbonate complexes and Cr hydroxides. Precipitation of CrIII in the form of colloidal particles of variable diameters was confirmed by nanoparticle (NP) tracking analysis, single particle ICP-MS, and single particle counting. In terms of time-weighted dissolved (< 0.22 μm) Cr concentration, CrIII was 4 to 10 times more toxic than CrVI. However, CrIII ecotoxicity could arise from interactions between free ionic CrIII and algae at the beginning of the test, from the presence of Cr-bearing NPs, or from a combination of the 2. Future ecotoxicological studies must pay more attention to Cr speciation to reliably compare the ecotoxicity of CrIII and CrVI. Environ Toxicol Chem 2018;37:983–992. © 2017 SETAC
Sewage sludge biochars management—Ecotoxicity, mobility of heavy metals, and soil microbial biomass
Authors: Mierzwa-Hersztek Monika , Gondek Krzysztof , Klimkowicz-Pawlas Agnieszka , Baran Agnieszka , Bajda Tomasz
Published: November 2017

Production of biochar from sewage sludge may be a promising solution for sewage sludge management and improvement of soil properties, including carbon dioxide sequestration. The aim of the present study was to compare the effects of biochars derived from different sewage sludges on soil physicochemical and biological properties, ecotoxicity, and plant yield. Three biochars (produced at a temperature of 300 °C) were applied into sandy acid soil in doses of 0.5, 1, and 2% (w/w). Depending on the type and dose, the application of sewage sludge biochars into the soil caused diverse effects on the parameters of soil biological activity (microbial biomass [Cmic], soil respiration, and value of metabolic quotient). No correlation between the applied dose of biochars and inhibition of Vibrio fischeri luminescence was observed. The factor with a stronger impact on the activity of V. fischeri was the type of biochar. The use of the OSTRACODTOXKIT F test (MicroBioTests) showed that the addition of sewage sludge biochar (regardless of its dose) reduced the soil toxicity to Heterocypris incongruens compared with the control soil. A significant increase of Poa pratensis L. biomass was obtained in soils with 1 and 2% additions of each of the biochars. The addition of biochars in doses of 1 and 2% to the soil had greater effect on the content of mobile forms of Cu, Pb, and Cd than the 0.5% dose compared with the control. Environ Toxicol Chem 2018;37:1197–1207. © 2017 SETAC

Abstract
Long‐term sediment yield from a small catchment in southern Brazil affected by land use and soil management changes
Authors: Minella Jean P.G. , Merten Gustavo H. , Barros Claúdia A.P. , Ramon Rafael , Schlesner Alexandre , Clarke Robin T. , Moro Michele , Dalbianco Leandro
Published: November 2017

Sediments produced from eroding cultivated land can cause on‐site and off‐site effects that cause considerable economic and social impacts. Despite the importance of soil conservation practices (SCP) for the control of soil erosion and improvements in soil hydrological functions, limited information is available regarding the effects of SCP on sediment yield (SY) at the catchment scale. This study aimed to investigate the long‐term relationships between SY and land use, soil management, and rainfall in a small catchment. To determine the effects of anthropogenic and climatic factors on SY, rainfall, streamflow, and suspended sediment concentration were monitored at 10‐min intervals for 14 years (2002–2016), and the land use and soil management changes were surveyed annually. Using a statistical procedure to separate the SY effects of climate, land use, and soil management, we observed pronounced temporal effects of land use and soil management changes on SY. During the first 2 years (2002–2004), the land was predominantly cultivated with tobacco under a traditional tillage system (no cover crops and ploughed soil) using animal traction. In that period, the SY reached approximately 400 t·km−2·year−1. From 2005 to 2009, a soil conservation programme introduced conservation tillage and winter cover crops in the catchment area, which lowered the SY to 50 t·km−2·year−1. In the final period (2010–2016), the SCP were partially abandoned by farmers, and reforested areas increased, resulting in an SY of 150 t·km−2·year−1. This study also discusses the factors associated with the failure to continue using SCP, including structural support and farmer attitudes.
Treatment of acquired Thrombotic Thrombocytopenic Purpura in the U.S. remains heterogeneous: Current and future points of clinical equipoise
Authors: Mazepa Marshall A. , Raval Jay S. , Brecher Mark E. , Park Yara A.
Published: November 2017

Background

The purpose of this survey was to describe current practices in the U.S. for treatment of acquired Thrombotic Thrombocytopenic Purpura (TTP), compare these with prior U.S. and current Canadian practices, and identify areas of clinical equipoise.

Study design and methods

A research team member administered the survey by telephone. Questions included an estimate of the annual patient volume treated, apheresis and medical therapy practices for acquired TTP.

Results

32 centers from 22 states were surveyed. ADAMTS13 activity is used for confirmation of the diagnosis of acquired TTP (97%). Most commonly, daily plasma exchange (therapeutic plasma exchange [TPE]) is initiated with plasma as replacement fluid (91%) at 1.0 Plasma Volume (72%) and stopped with a platelet count of 150 × 109/L (66%), and then TPE is tapered off (69%). Compared with a U.S. survey from 1998, a greater proportion of centers use plasma exclusively as the replacement fluid exclusively (29/32 vs 2/14 in 1998; P < .0001) and taper TPE (22/32 vs 8/20 in 1998, P = .0499). Compared with Canadian survey in 2016, a greater proportion of U.S. centers use plasma over cryosupernatant (29/32 vs 2/13 CAG centers, P < .0001) and initiate TPE with 1.0 PV compared with 1.5 PV (23/32 vs 0/14 CAG centers, P < .0001). Corticosteroid use is common but not universal (U.S. and CAG) and use of rituximab heterogeneous.

Conclusion

Treatment of acquired TTP in the U.S. remains heterogeneous. Points of clinical equipoise identified were PV exchanged (1.0 vs >1.0), tapering of TPE versus none, and rituximab use.
Seeing is believing: A review of apheresis therapy in the treatment of ophthalmologic disease
Authors: Graham Brendan C. , Pulido Jose S. , Winters Jeffrey L.
Published: November 2017

Apheresis procedures have a role in treatment of disparate diseases involving many different organ systems. Often the disease processes where apheresis plays a role in treatment are considered “orphan diseases”—relatively rare disease processes that lack specific pharmaceutical agents or established treatment protocols. Many of these disease processes can affect the eye with devastating results for the eyesight of these patients. The unique ability of apheresis to affect disease by modifying blood plasma and modulating disease‐causing agents therein renders apheresis procedures valuable tools in the treatment of certain ophthalmologic diseases. This review comprehensively evaluates the role of apheresis in the treatment of ophthalmologic diseases of the eye and surrounding orbit including age‐related macular degeneration, bilateral diffuse uveal melanocytic proliferation, paraneoplastic retinopathy, atopic keratoconjunctivitis, sympathetic ophthalmia, and endocrine‐associated ophthalmopathy. Apheresis procedure parameters are provided for the apheresis practitioner based on review of the relevant literature.
Development of spin‐orbit coupling for stochastic configuration interaction techniques
Authors: Murphy Paul , Coe Jeremy P. , Paterson Martin J.
Published: November 2017

To perform spin‐orbit coupling calculations on atoms and molecules, good zeroth‐order wavefunctions are necessary. Here, we present the software development of the Monte Carlo Configuration Interaction (MCCI) method, to enable calculation of such properties, where MCCI iteratively constructs a multireference wavefunction using a stochastic procedure. In this initial work, we aim to establish the efficacy of this technique in predicting the splitting of otherwise degenerate energy levels on a range of atoms and small diatomic molecules. It is hoped that this work will subsequently act as a gateway toward using this method to investigate singlet‐triplet interactions in larger multireference molecules. We show that MCCI can generate very good results using highly compact wavefunctions compared to other techniques, with no prior knowledge of important orbitals. Higher‐order relativistic effects are neglected and spin‐orbit coupling effects are incorporated using first‐order degenerate perturbation theory with the Breit‐Pauli Hamiltonian and effective nuclear charges in the one‐electron operator. Results are obtained and presented for B, C, O, F, Si, S, and Cl atoms and OH, CN, NO, and C2 diatomic radicals including spin‐orbit coupling constants and the relative splitting of the lowest energy degenerate state for each species. Convergence of MCCI to the full configuration interaction result is demonstrated on the multireference problem of stretched OH. We also present results from the singlet‐triplet interaction between the
and both the
and
states of the O2 molecule. © 2017 Wiley Periodicals, Inc.
A reference‐free stockholder partitioning method based on the force on electrons
Authors: Fias Stijn , Heidar‐Zadeh Farnaz , Anderson James S. M. , Ayers Paul W. , Parr Robert G.
Published: November 2017

We argue that when one divides a molecular property into atom‐in‐a‐molecule contributions, one should perform the division based on the property density of the quantity being partitioned. This is opposition to the normal approach, where the electron density is given a privileged role in defining the properties of atoms‐in‐a‐molecule. Because partitioning each molecular property based on its own property density is inconvenient, we design a reference‐free approach that does not (directly) refer atomic property densities. Specifically, we propose a stockholder partitioning method based on relative influence of a molecule's atomic nuclei on the electrons at a given point in space. The resulting method does not depend on an “arbitrary” choice of reference atoms and it has some favorable properties, including the fact that all of the electron density at an atomic nucleus is assigned to that nucleus and the fact all the atoms in a molecule decay at a uniform asymptotic rate. Unfortunately, the resulting model is not easily applied to spatially degenerate ground states. Furthermore, the practical realizations of this strategy that we tried here gave disappointing numerical results. © 2017 Wiley Periodicals, Inc.
EXAT: EXcitonic analysis tool
Authors: Jurinovich Sandro , Cupellini Lorenzo , Guido Ciro A. , Mennucci Benedetta
Published: November 2017

We introduce EXcitonic Analysis Tool (EXAT), a program able to compute optical spectra of large excitonic systems directly from the output of quantum mechanical calculations performed with the popular Gaussian 16 package. The software is able to combine in an excitonic scheme the single‐chromophore properties and exciton couplings to simulate energies, coefficients, and excitonic spectra (UV‐vis, CD, and LD). The effect of the environment can also be included using a Polarizable Continuum Model. EXAT also presents a simple graphical user interface, which shows on‐screen both site and exciton properties. To show the potential of the method, we report two applications on a a chiral perturbed BODIPY system and DNA G‐quadruplexes, respectively. The program is available online at http://molecolab.dcci.unipi.it/tools/. © 2017 Wiley Periodicals, Inc.
Generation of insulin‐producing cells from human adipose‐derived mesenchymal stem cells on PVA scaffold by optimized differentiation protocol
Authors: Enderami Seyed Ehsan , Soleimani Masoud , Mortazavi Yousef , Nadri Samad , Salimi Ali
Published: November 2017

The studies have been done on patient‐specific human adipose‐derived from mesenchymal stem cells (hADSCs) like a series of autologous growth factors and nanofibrous scaffolds (3D culture) will probably have many benefits for regenerative medicine in type 1 diabetes mellitus (TIDM) patients in the future. For this purpose, we established a polyvinyl alcohol (PVA) scaffold and a differentiation protocol by adding platelet‐rich plasma (PRP) that induces the hADSCs into insulin‐producing cells (IPCs). The characteristics of the derived IPCs in 3D culture were compared with conventional culture (2D) groups evaluated at the mRNA and protein levels. The viability of induced pancreatic cells was 14 days. The in vitro studies showed that the treatment of hADSCs in the 3D culture resulted in differentiated cells with strong characteristics of IPCs including pancreatic‐like cells, the expression of the islet‐associated genes at the mRNA and protein levels in comparison of 2D culture group. Furthermore, the immunoassay tests showed that these differentiated cells in these two groups are functional and secreted C‐peptide and insulin in a glucose stimulation challenge. The results of our study for the first time demonstrated that the PVA nanofibrous scaffolds along with the optimized differentiation protocol with PRP can enhance the differentiation of IPCs from hADSCs. In conclusion, this study provides a new approach to the future pancreatic tissue engineering and beta cell replacement therapies for T1DM.
Tumor‐suppressive roles of ΔNp63β‐miR‐205 axis in epithelial–mesenchymal transition of oral squamous cell carcinoma via targeting ZEB1 and ZEB2
Authors: Hashiguchi Yuma , Kawano Shintaro , Goto Yuichi , Yasuda Kaori , Kaneko Naoki , Sakamoto Taiki , Matsubara Ryota , Jinno Teppei , Maruse Yasuyuki , Tanaka Hideaki , Morioka Masahiko , Hattori Taichi , Tanaka Shoichi , Kiyoshima Tamotsu , Nakamura Seiji
Published: November 2017

We previously revealed that epithelial‐to‐mesenchymal transition (EMT) was mediated by ΔNp63β, a splicing variant of ΔNp63, in oral squamous cell carcinoma (OSCC). Recent studies have highlighted the involvement of microRNA (miRNA) in EMT of cancer cells, though the mechanism remains unclear. To identify miRNAs responsible for ΔNp63β‐mediated EMT, miRNA microarray analyses were performed by ΔNp63β‐overexpression in OSCC cells; SQUU‐B, which lacks ΔNp63 expression and displays EMT phenotypes. miRNAs microarray analyses revealed miR‐205 was the most up‐regulated following ΔNp63β‐overexpression. In OSCC cells, miR‐205 expression was positively associated with ΔNp63 and negatively with zinc‐finger E‐box binding homeobox (ZEB) 1 and ZEB2, potential targets of miR‐205. miR‐205 overexpression by miR‐205 mimic transfection into SQUU‐B cells led to decreasing ZEB1, ZEB2, and mesenchymal markers, increasing epithelial markers, and reducing cell motilities, suggesting inhibition of EMT phenotype. Interestingly, the results opposite to this phenomenon were obtained by transfection of miR‐205 inhibitor into OSCC cells, which express ΔNp63 and miR‐205. Furthermore, target protector analyses revealed direct regulation by miR‐205 of ZEB1 and ZEB2 expression. These results showed tumor‐suppressive roles of ΔNp63β and miR‐205 by inhibiting EMT thorough modulating ZEB1 and ZEB2 expression in OSCC.
Regulation of calcification in human aortic smooth muscle cells infected with high‐glucose‐treated Porphyromonas gingivalis
Authors: Chen Te‐Chuan , Lin Chien‐Tsong , Chien Shao‐Ju , Chang Shun‐Fu , Chen Cheng‐Nan
Published: November 2017

Porphyromonas (P.) gingivalis infection leading to the periodontitis has been associated with the development of systemic diseases, including cardiovascular diseases and diabetes. However, the effect of a high concentration of glucose (HG) on the invasion efficiency of P. gingivalis and the consequent modulation of pathogenesis in vascular cells, especially in the vascular smooth muscle cells (VSMCs), remains unclear. Hence, the aim of this study was to investigate whether treating P. gingivalis with HG could change its invasion capability and result in VSMC calcification and the underlying mechanism. Human aortic SMCs (HASMCs) and P. gingivalis strain CCUG25226 were used in this study. We found that HGPg infection of HASMCs could initiate the HASMC calcification by stimulating the autocrine regulation of bone morphogenetic protein (BMP) 4 in HASMCs. The upregulation of BMP4 expression in HASMCs was mediated by toll‐like receptor 4 and ERK1/2‐p38 signaling after P. gingivalis infection. Moreover, the autocrine action of BMP4 in HGPg infection‐initiated HASMC calcification upregulated BMP4‐specific downstream smad1/5/8‐runx2 signaling to increase the expressions of bone‐related matrix proteins, that is, osteopontin, osteocalcin, and alkaline phosphatase. This study elucidates the detailed mechanism of HGPg infection‐initiated calcification of HASMCs and indicates a possible therapeutic role of BMP4 in P. gingivalis infection‐associated vascular calcification.
NONO ubiquitination is mediated by FBW7 and GSK3 β via a degron lost upon chromosomal rearrangement in cancer
Authors: Alfano Luigi , Caporaso Antonella , Altieri Angela , Costa Caterina , Forte Iris M. , Iannuzzi Carmelina A. , Barone Daniela , Esposito Luca , Giordano Antonio , Pentimalli Francesca
Published: November 2017

NONO is an RNA‐binding protein involved in transcription, mRNA splicing, DNA repair, and checkpoint activation in response to UV radiation. NONO expression has been found altered in several tumor types, including prostate, colon, breast, melanoma, and in papillary renal carcinoma, in which an X chromosome inversion generates a NONO‐TFE3 fusion protein. Upon such rearrangement, NONO loses its C‐terminal domain. Through bioinformatics analysis, we identified a putative degron motif, known to be recognized by the Skp1‐Cul1‐F‐box‐protein (SCF) complex. Here, we evaluated how this domain could affect NONO protein biology. We showed that NONO interacts with the nuclear FBW7α isoform and its ubiquitination is regulated following modulation of the GSK3β kinase. Mutation of T428A/T432A within the degron impaired polyubiquitination upon FBW7α and GSK3β overexpression. Overall, our data suggest that NONO is likely subjected to proteasome‐mediated degradation and add NONO to the list of proteins targeted by FBW7, which is itself often deregulated in cancer.
Methylation‐associated DOK1 and DOK2 down‐regulation: Potential biomarkers for predicting adverse prognosis in acute myeloid leukemia
Authors: He Pin‐Fang , Xu Zi‐Jun , Zhou Jing‐Dong , Li Xi‐Xi , Zhang Wei , Wu De‐Hong , Zhang Zhi‐Hui , Lian Xin‐Yue , Yao Xin‐Yu , Deng Zhao‐Qun , Lin Jiang , Qian Jun
Published: November 2017

DOK‐1 and DOK‐2 (DOK1/2) are closely related members of downstream of tyrosine kinase (DOK) family genes, which are found to be frequently rearranged in several hematopoietic cancers. However, the clinical implications of DOK1/2 in acute myeloid leukemia (AML) remain largely unknown. To investigate the clinical significance, real‐time quantitative PCR (RQ‐PCR) was carried out to detect DOK1/2 expressions in 125 de novo AML patients and 28 healthy controls. Real‐time quantitative methylation‐specific PCR (RQ‐MSP) and bisulfite sequencing PCR (BSP) were applied to detect DOK1/2 methylation level and density. DOK1/2 expressions were significantly down‐regulated in AML patients. The promoters of DOK1/2 were highly hypermethylated and negatively correlated with DOK1/2 expressions in AML patients. In addition, we also confirmed that DOK1/2 expressions could be restored by DOK1/2 demethylation using 5‐aza‐2′‐deoxycytidine in leukemia cell line THP‐1. Survival analyses showed that low‐expressed DOK1/2 were associated with markedly shorter overall survival and leukemia free survival in both whole‐cohort AML and non‐M3 AML patients. Multivariate analyses further revealed that DOK1/2 were act as independent prognostic factors in AML patients. These findings indicate that decreased DOK1/2 expressions associated with their promoter hypermethylations predict adverse prognosis in AML.
Retracted: MicroRNA‐497 accelerates apoptosis while inhibiting proliferation, migration, and invasion through negative regulation of the MAPK/ERK signaling pathway via RAF‐1
Authors: Tao Ling , Zhang Chun‐Yu , Guo Lei , Li Xia , Han Na‐Na , Zhou Qi , Liu Zhi‐Le
Published: November 2017

The aim of this study is to explore the various modes of action miR‐497 has on human cervical cancer (CC) cell behavior. We also speculate that miR‐497 achieves its anti‐tumor role by governing RAF‐1 via MAPK/ERK signaling pathway. CC tissues with corresponding adjacent normal tissues were collected from 168 CC patients. RAF‐1‐positive cells were identified by means of immunohistochemistry in tissues. A series of inhibitors, mimics and siRNA against RAF‐1 were introduced to validate regulatory mechanisms for miR‐497 and RAF‐1. Quantitative real‐time polymerase chain reaction (qRT‐PCR) and Western blot assay were employed for evaluating alternations of miR‐497, RAF‐1, and MAPK/ERK signaling pathway. HeLa cell proliferation, invasion, migration, cycle progression, and apoptosis were assessed by means of CCK‐8, wound‐healing, transwell invasion assays, and flow cytometry, respectively. The target prediction program and luciferase activity determination were used to identify miR‐497 targeting RAF‐1. We determined reduced miR‐497 expression and elevated expression of RAF‐1 in CC tissues as opposed to adjacent tissues. Transfection of miR‐497 mimics and siRNA‐RAF‐1 both decreased levels of MEK1, ERK1, and p38 phosphorylation in HeLa cells, inhibited cell proliferation, migration and invasion, induced more cells arrested in the G0/G1 phase, and promoted cell apoptosis; while miR‐497 inhibitors led to opposite results. These findings indicate miR‐497 as a tumor suppressor results from negative regulation of the MAPK/ERK signaling pathway via RAF‐1 in CC.
Toll like receptor signaling pathway as a potential therapeutic target in colorectal cancer
Authors: Moradi‐Marjaneh Reyhaneh , Hassanian Seyed Mahdi , Fiuji Hamid , Soleimanpour Saman , Ferns Gordon A. , Avan Amir , Khazaei Majid
Published: November 2017

Toll like receptor (TLR) signaling is involved in activating innate and adaptive immune responses and plays a critical role in inflammation‐induced diseases such as colorectal cancer (CRC). Dysregulation of this signaling pathway can result in disturbance of epithelial layer hemostasis, chronic inflammatory, excessive repair responses, and development of CRC. There is now substantial evidence for the benefit of targeting of this pathway in cancer treatment, and several agents have been approved, such as BCG (Bacillus Calmette Guérin), MPL (monophosphoryl lipid A) and imiquimod. This review summarizes the current knowledge about the different functions of TLRs on tumor cells and their application in cancer therapy with particular emphasis on recent preclinical and clinical research in treatment of CRC.
miR‐34b‐5p inhibition attenuates lung inflammation and apoptosis in an LPS‐induced acute lung injury mouse model by targeting progranulin
Authors: Xie Wang , Lu Qingchun , Wang Kailing , Lu Jingjing , Gu Xia , Zhu Dongyi , Liu Fanglei , Guo Zhongliang
Published: November 2017

Inflammation and apoptosis play important roles in the initiation and progression of acute lung injury (ALI). Our previous study has shown that progranulin (PGRN) exerts lung protective effects during LPS‐induced ALI. Here, we have investigated the potential roles of PGRN‐targeting microRNAs (miRNAs) in regulating inflammation and apoptosis in ALI and have highlighted the important role of PGRN. LPS‐induced lung injury and the protective roles of PGRN in ALI were first confirmed. The function of miR‐34b‐5p in ALI was determined by transfection of a miR‐34b‐5p mimic or inhibitor in intro and in vivo. The PGRN level gradually increased and subsequently significantly decreased, reaching its lowest value by 24 hr; PGRN was still elevated compared to the control. The change was accompanied by a release of inflammatory mediators and accumulation of inflammatory cells in the lungs. Using bioinformatics analysis and RT‐PCR, we demonstrated that, among 12 putative miRNAs, the kinetics of the miR‐34b‐5p levels were closely associated with PGRN expression in the lung homogenates. The gain‐ and loss‐of‐function analysis, dual‐luciferase reporter assays, and rescue experiments confirmed that PGRN was the functional target of miR‐34b‐5p. Intravenous injection of miR‐34b‐5p antagomir in vivo significantly inhibited miR‐34b‐5p up‐regulation, reduced inflammatory cytokine release, decreased alveolar epithelial cell apoptosis, attenuated lung inflammation, and improved survival by targeting PGRN during ALI. miR‐34b‐5p knockdown attenuates lung inflammation and apoptosis in an LPS‐induced ALI mouse model by targeting PGRN. This study shows that miR‐34b‐5p and PGRN may be potential targets for ALI treatments.
A novel epididymal quiescence factor inhibits sperm motility by modulating NOS activity and intracellular NO‐cGMP pathway
Authors: Ghosh Prasanta , Mukherjee Sandipan , Bhoumik Arpita , Dungdung Sandhya R.
Published: November 2017

Mature and potentially motile spermatozoa stored in cauda epididymis in an inactive state for approximately 30 days; however, during ejaculation they regain motility. To understand the actual molecular mechanism of the sperm quiescence during caudal stay, a proteinaceous quiescence factor (QF) has been purified from caprine epididymal plasma to apparent homogeneity. In the present study complete purification, detailed characterization as well as mechanistic pathway of QF has been described. QF is purified to 215‐fold with 45% activity recovery. It is a 59 kDa monomeric protein with isoelectric point 5.8 and optimally active at pH 7.5. Circular dichroism spectroscopy and atomic force microscopy study confirm its α‐helical secondary structure and globular tertiary conformation. QF is a thermo‐stable protein as higher temperature does not alter its helical structure. N‐terminal amino acid sequencing and MALDI analysis of QF did not find 100% similarity with any available protein of the database, proved its novelty. QF at 2 μM dose inhibits sperm progressive forward motility within 10 min. This motility inhibitory activity of QF is mediated by reducing NOS enzyme activity and subsequently decreasing the intracellular NO and cGMP concentration. It does not modulate intracellular Ca++ and cAMP concentration. QF has no adverse effect on DNA integrity and morphology of spermatozoa. Motility inhibitory action of QF is reversible. Thus, the role of QF in maintaining energy saving quiescence state of mature cauda spermatozoa and its reactive nitrogen species reducing activity may lead to a new direction for storage of spermatozoa and idiopathic male infertility.
LncRNA‐TCONS_00034812 in cell proliferation and apoptosis of pulmonary artery smooth muscle cells and its mechanism
Authors: Liu Yun , Sun Zengxian , Zhu Jinquan , Xiao Bingxin , Dong Jie , Li Xiaomin
Published: November 2017

Long noncoding RNAs (lncRNAs) have been discovered to be playing important role in various biological processes. However, the contribution of lncRNAs to pulmonary artery hypertension (PAH) remains largely unknown. Pulmonary vascular remodeling is an important pathological feature of PAH, leading to increased vascular resistance and reduced compliance. Here, we investigated the biological role of lncRNAs in PAH. Differences in the lncRNAs and mRNAs between hypoxia PAH rats and normoxia rats were screened using microarray analysis. The results showed that 36 lncRNAs and 519 mRNAs were upregulated in the pulmonary arteries (PAs) of hypoxia PAH rats, whereas 111 lncRNAs and 246 mRNAs were downregulated. Expressions of the screened lncRNAs, including TCONS_00034812, were validated by real‐time PCR. We revealed that the expression of TCONS_00034812 was significantly downregulated in PAs of PAH rats and hypoxia pulmonary artery smooth muscle cells (PASMCs). TCONS_00034812 knockdown promoted proliferation and inhibited apoptosis of PASMCs in vitro. Moreover, TCONS_00034812 regulated PASMCs function in vitro. We found that TCONS_00034812 increased the expression of transcription factors Stox1. TCONS_00034812 and Stox1 knockdown mediated PASMCs function through MAPK signaling. Our findings imply lncRNA as a critical regulator in PAH and demonstrate the potential of gene therapy and drug development for treating PAH. The present study reveals a novel mechano responsive lncRNA‐TCONS_00034812, which modulates PASMCs proliferation and apoptosis, and participates in vascular remodeling during PAH.
Development of an in vivo mouse model of discogenic low back pain
Authors: Shi Changgui , Das Vaskar , Li Xin , Kc Ranjan , Qiu Sujun , O‐Sullivan InSug , Ripper Richard L. , Kroin Jeffrey S. , Mwale Fackson , Wallace Atiyayein A. , Zhu Bingqian , Zhao Lan , van Wijnen Andre J. , Ji Mingliang , Lu Jun , Votta‐Velis Gina , Yuan Wen , Im Hee‐Jeong
Published: November 2017

Discogenic low back pain (DLBP) is extremely common and costly. Effective treatments are lacking due to DLBP's unknown pathogenesis. Currently, there are no in vivo mouse models of DLBP, which restricts research in this field. The aim of this study was to establish a reliable DLBP model in mouse that captures the pathological changes in the disc and allows longitudinal pain testing. The model was generated by puncturing the mouse lumbar discs (L4/5, L5/6, and L6/S1) and removing the nucleus pulposus using a microscalpel under the microscope. Histology, molecular pathways, and pain‐related behaviors were examined. Over 12 weeks post‐surgery, animals displayed the mechanical, heat, and cold hyperalgesia along with decreased burrowing and rearing. Histology showed progressive disc degeneration with loss of disc height, nucleus pulposus reduction, proteoglycan depletion, and annular fibrotic disorganization. Immunohistochemistry revealed a substantial increase in inflammatory mediators at 2 and 4 weeks. Nerve growth factor was upregulated from 2 weeks to the end of the experiment. Nerve fiber ingrowth was induced in the injured discs after 4 weeks. Disc‐puncture also produced an upregulation of neuropeptides in dorsal root ganglia neurons and an activation of glial cells in the spinal cord dorsal horn. These findings indicate that the cellular and structural changes in discs, as well as peripheral and central nervous system plasticity, paralleled persistent, and robust behavioral pain responses. Therefore, this mouse DLBP model could be used to investigate mechanisms underlying discogenic pain, thereby facilitating effective drug screening and development of treatments for DLBP.
Muscle regeneration potential and satellite cell activation profile during recovery following hindlimb immobilization in mice
Authors: Guitart Maria , Lloreta Josep , Mañas‐Garcia Laura , Barreiro Esther
Published: November 2017

Reduced muscle activity leads to muscle atrophy and function loss in patients and animal models. Satellite cells (SCs) are postnatal muscle stem cells that play a pivotal role in skeletal muscle regeneration following injury. The regenerative potential, satellite cell numbers, and markers during recovery following immobilization of the hindlimb for 7 days were explored. In mice exposed to 7 days of hindlimb immobilization, in those exposed to recovery (7 days, splint removal), and in contralateral control muscles, muscle precursor cells were isolated from all hindlimb muscles (fluorescence‐activated cell sorting, FACS) and SCs, and muscle regeneration were identified using immunofluorescence (gastrocnemius and soleus) and electron microscopy (EM, gastrocnemius). Expression of ki67, pax7, myoD, and myogenin was quantified (RT‐PCR) from SC FACS yields. Body and grip strength were determined. Following 7 day hindlimb immobilization, a decline in SCs (FACS, immunofluorescence) was observed together with an upregulation of SC activation markers and signs of muscle regeneration including fusion to existing myofibers (EM). Recovery following hindlimb immobilization was characterized by a program of muscle regeneration events. Hindlimb immobilization induced a decline in SCs together with an upregulation of markers of SC activation, suggesting that fusion to existing myofibers takes place during unloading. Muscle recovery induced a significant rise in muscle precursor cells and regeneration events along with reduced SC activation expression markers and a concomitant rise in terminal muscle differentiation expression. These are novel findings of potential applicability for the treatment of disuse muscle atrophy, which is commonly associated with severe chronic and acute conditions.
MicroRNAs as predictive biomarkers for myocardial injury in aged mice following myocardial infarction
Authors: Qipshidze Kelm Natia , Piell Kellianne M. , Wang Eugenia , Cole Marsha P.
Published: November 2017

The occurrence of myocardial infarction (MI) increases appreciably with age. In the Framingham Heart Study, the incidence of MI more than doubles for men and increases more than five‐fold in women (ages 55–64 years compared to 85–94 years). MicroRNAs (miRNAs) quantitatively regulate their target's expression post‐transcriptionally by either silencing action through binding at the 3′UTR domains or degrading the messages at their coding regions. In either case, these regulations affect the cardiac transcriptional output and cardiac function. Among the known cardiac associated miRNA, miRNA‐1, miRNA‐133a, and miRNA‐34a have been shown to induce adverse structural remodeling to impair cardiac contractile function. In the present study, an in vivo model of MI in young (3 month) and old (22 month) mice is used to investigate the possible role whereby these three miRNAs exert negative effects on heart function following MI. Herein we demonstrate that in older mouse heart, all three microRNAs show increased levels of expression, while miRNA‐1 shows a further increase in old mouse heart following MI, which corresponds to left ventricular (LV) wall thinning. These structural changes in cardiac tissue may causes downstream LV dilation and subsequent LV dysfunction. Results presented here suggest that significantly elevated levels of miRNA‐1 in post‐MI old heart could be predictive of cardiac injury in older mice as the high risk biomarker for MI in older individuals.
Inhibition of miR‐200b/miR‐429 contributes to neuropathic pain development through targeting zinc finger E box binding protein‐1
Authors: Yan Xue‐Tao , Zhao Ying , Cheng Xiao‐Li , He Xiang‐Hu , Wang Yu , Zheng Wen‐Zhong , Chen Hu , Wang Yan‐Lin
Published: November 2017

Many studies have reported that microRNAs participate in neuropathic pain development. Previously, miR‐200b and miR‐429 are reported to be involved in various diseases. In our current study, we focused on their roles in neuropathic pain and we found that miR‐200b and miR‐429 were significantly decreased in chronic constriction injury (CCI) rat spinal cords and isolated microglials. miR‐200b and miR‐429 overexpression were able to relieve neuropathic pain through modulating PWT and PWL in CCI rats. Meanwhile, we observed that both miR‐200b and miR‐429 upregulation could repress neuroinflammation via inhibiting inflammatory cytokines such as IL‐6, IL‐1β, and TNF‐α in CCI rats. By carry out bioinformatics technology, Zinc finger E box binding protein‐1 (ZEB1) was predicted as target of miR‐200b, and miR‐429 and dual‐luciferase reporter assays confirmed the correlation between them. ZEB1 has been reported to regulate a lot of diseases. Here, we found that ZEB1 was greatly increased in CCI rats and miR‐200b and miR‐429 overexpression markedly suppressed ZEB1 mRNA expression in rat microglial cells. In addition, knockdown of ZEB1 can reduce neuropathic pain development and co‐transfection of LV‐anti‐miR‐200b/miR‐429 reversed this phenomenon in vivo. Taken these together, our results suggested that miR‐200b/miR‐429 can serve as an important regulator of neuropathic pain development by targeting ZEB1.
Physiological role of urothelial cancer‐associated one long noncoding RNA in human skeletogenic cell differentiation
Authors: Ishikawa Takanori , Nishida Takashi , Ono Mitsuaki , Takarada Takeshi , Nguyen Ha Thi , Kurihara Shinnosuke , Furumatsu Takayuki , Murase Yurika , Takigawa Masaharu , Oohashi Toshitaka , Kamioka Hiroshi , Kubota Satoshi
Published: November 2017

A vast number of long‐noncoding RNAs (lncRNA) are found expressed in human cells, which RNAs have been developed along with human evolution. However, the physiological functions of these lncRNAs remain mostly unknown. In the present study, we for the first time uncovered the fact that one of such lncRNAs plays a significant role in the differentiation of chondrocytes and, possibly, of osteoblasts differentiated from mesenchymal stem cells, which cells eventually construct the human skeleton. The urothelial cancer‐associated 1 (UCA1) lncRNA is known to be associated with several human malignancies. Firstly, we confirmed that UCA1 was expressed in normal human chondrocytes, as well as in a human chondrocytic cell line; whereas it was not detected in human bone marrow mesenchymal stem cells (hBMSCs). Of note, although UCA1 expression was undetectable in hBMSCs, it was markedly induced along with the differentiation toward chondrocytes, suggesting its critical role in chondrogenesis. Consistent with this finding, silencing of the UCA1 gene significantly repressed the expression of chondrogenic genes in human chondrocytic cells. UCA1 gene silencing and hyper‐expression also had a significant impact on the osteoblastic phenotype in a human cell line. Finally, forced expression of UCA1 in a murine chondrocyte precursor, which did not possess a UCA1 gene, overdrove its differentiation into chondrocytes. These results indicate a physiological and important role of this lncRNA in the skeletal development of humans, who require more sustained endochondral ossification and osteogenesis than do smaller vertebrates.
Skeletal muscle performance and ageing
Authors: Tieland Michael , Trouwborst Inez , Clark Brian C.
Published: November 2017

The world population is ageing rapidly. As society ages, the incidence of physical limitations is dramatically increasing, which reduces the quality of life and increases healthcare expenditures. In western society, ~30% of the population over 55 years is confronted with moderate or severe physical limitations. These physical limitations increase the risk of falls, institutionalization, co‐morbidity, and premature death. An important cause of physical limitations is the age‐related loss of skeletal muscle mass, also referred to as sarcopenia. Emerging evidence, however, clearly shows that the decline in skeletal muscle mass is not the sole contributor to the decline in physical performance. For instance, the loss of muscle strength is also a strong contributor to reduced physical performance in the elderly. In addition, there is ample data to suggest that motor coordination, excitation–contraction coupling, skeletal integrity, and other factors related to the nervous, muscular, and skeletal systems are critically important for physical performance in the elderly. To better understand the loss of skeletal muscle performance with ageing, we aim to provide a broad overview on the underlying mechanisms associated with elderly skeletal muscle performance. We start with a system level discussion and continue with a discussion on the influence of lifestyle, biological, and psychosocial factors on elderly skeletal muscle performance. Developing a broad understanding of the many factors affecting elderly skeletal muscle performance has major implications for scientists, clinicians, and health professionals who are developing therapeutic interventions aiming to enhance muscle function and/or prevent mobility and physical limitations and, as such, support healthy ageing.
Near‐infrared diffuse reflectance signals for monitoring spreading depolarizations and progression of the lesion in a male rat focal cerebral ischemia model
Authors: Kawauchi Satoko , Nishidate Izumi , Nawashiro Hiroshi , Sato Shunichi
Published: November 2017

In ischemic stroke research, a better understanding of the pathophysiology and development of neuroprotection methods are crucial, for which in vivo imaging to monitor spreading depolarizations (SDs) and evolution of tissue damage is desired. Since these events are accompanied by cellular morphological changes, light‐scattering signals, which are sensitive to cellular and subcellular morphology, can be used for monitoring them. In this study, we performed transcranial imaging of near‐infrared (NIR) diffuse reflectance at ∼800 nm, which sensitively reflects light‐scattering change, and examined how NIR reflectance is correlated with simultaneously measured cerebral blood flow (CBF) for a rat middle cerebral artery occlusion (MCAO) model. After MCAO, wavelike NIR reflectance changes indicating occurrence of SDs were generated and propagated around the ischemic core for ∼90 min, during which time NIR reflectance increased not only within the ischemic core but also in the peripheral region. The area with increased reflectance expanded with increase in the number of SD occurrences, the correlation coefficient being 0.7686 (n = 5). The area with increased reflectance had become infarcted at 24 hr after MCAO. The infarct region was found to be associated with hypoperfusion or no‐flow response to SD, but hyperemia or hypoperfusion followed by hyperemia response to SD was also observed, and the regional heterogeneity seemed to be connected with the rat cerebrovasculature and hence existence/absence of collateral flow. The results suggest that NIR reflectance signals depicted early evolution of tissue damage, which was not seen by CBF changes, and enabled lesion progression monitoring in the present stroke model.
New algorithm for simulation of proteoglycan loss and collagen degeneration in the knee joint: Data from the osteoarthritis initiative
Authors: Mononen Mika E , Tanska Petri , Isaksson Hanna , Korhonen Rami K.
Published: November 2017

Osteoarthritis is a harmful joint disease but prediction of disease progression is problematic. Currently, there is only one modeling framework which can be applied to predict the progression of knee osteoarthritis but it only considers degenerative changes in the collagen fibril network. Here, we have developed the framework further by considering all of the major tissue changes (proteoglycan content, fluid flow, and collagen fibril network) occurring in osteoarthritis. While excessive levels of tissue stresses controlled degeneration of the collagen fibril network, excessive levels of tissue strains controlled the decrease in proteoglycan content and the increase in permeability. We created four knee joint models with increasing degrees of complexity based on the depth‐wise composition. Models were tested for normal and abnormal, physiologically relevant, loading conditions in the knee. Finally, the predicted depth‐wise compositional changes from each model were compared against experimentally observed compositional changes in vitro. The model incorporating the typical depth‐wise composition of cartilage produced the best match with experimental observations. Consistent with earlier in vitro experiments, this model simulated the greatest proteoglycan depletion in the superficial and middle zones, while the collagen fibril degeneration was located mostly in the superficial zone. The presented algorithm can be used for predicting simultaneous collagen degeneration and proteoglycan loss during the development of knee osteoarthritis. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1673–1683, 2018.
Rapid and green determination of 58 fragrance allergens in plush toys
Authors: Wang Zhijuan , Zhang Qing , Li Haiyu , Lv Qing , Wang Wan , Bai Hua
Published: November 2017

This paper presents a simple, rapid, and green method based on a static headspace gas chromatography–mass spectrometry for determining 58 fragrance allergens in plush toys. This study is the first to meet the requirement for simultaneously determining 66 fragrances, except for eight natural extracts restricted by the European Toy Safety Directive 2009/48/EC. A minimal amount of sample (20 mg) and acetone solvent (20 μL) were placed in a headspace vial. A gas–solid equilibration of fragrances between the headspace and the sample was achieved within 10 min under the vapor atmosphere of acetone at 200°C, which allowed the fragrances in the sample to be measured by gas chromatography–mass spectrometry. Results showed that the interference caused by different sample matrices was negligible. The proposed method exhibited determination results that were highly similar to those of traditional ultrasonic extraction with sufficient sensitivity (limits of quantification: 0.02–10 mg/kg) for 58 fragrances, indicating its accuracy and reliability. The average recoveries ranged from 71.3 to 137.4%, and the relative standard deviation (n = 6) varied from 1.1 to 18.0%. Finally, the method was applied to monitor the fragrances in 20 commercial toys. This study provides a good reference for rapidly and greenly determining the semi‐volatile analytes in toys, textiles, and other products.
Application of high‐speed counter‐current chromatography and HPLC to separate and purify of three polyacetylenes from Platycodon grandiflorum
Authors: Chen Bao , Liu Zhengbo , Zhang Yuwei , Li Wei , Sun Yanjun , Wang Yufang , Wang Yanhua , Sun Yinshi
Published: November 2017

Three polyacetylenes were isolated and purified from Platycodon grandiflorum A. DC for the first time by high‐speed counter‐current chromatography using a two‐phase solvent system composed of hexane/ethyl acetate/methanol/water (1:31:1:31, v/v/v/v) and high‐performance liquid chromatography with an Agilent ZORBAX® SB‐C18 column (4.6 mm × 150 mm, 5 μm). After separation by high‐speed counter‐current chromatography and high‐performance liquid chromatography, we obtained 3.5 mg of platetyolin A, 4.1 mg of platetyolin B, and 18.1 mg of lobetyolin with purities of 97.2, 96.7, and 96.9%, respectively. The purity of each compound was assessed by high‐performance liquid chromatography and the chemical structures were evaluated by high‐resolution electrospray ionization time‐of‐flight mass spectrometry and one‐ and two‐dimensional NMR spectroscopy. Among the isolated compounds, platetyolin A and platetyolin B are newly reported compounds.