bluuebunny/crossref_metadata_2025 huggingface dataset:
Stathmin inhibits proliferation and differentiation of dental pulp stem cells via sonic hedgehog/Gli
Authors: Ma Dandan , Yu Haiyue , Xu Shuaimei , Wang He , Zhang Xiaoyi , Ning Tingting , Wu Buling
Published: April 2018

The mineralization of dental pulp stem cells is an important factor in the tissue engineering of teeth, but the mechanism is not yet obvious. This study aimed to identify the effect of Stathmin on the proliferation and osteogenic/odontoblastic differentiation of human dental pulp stem cells (hDPSCs) and to explore whether the Shh signalling pathway was involved in this regulation. First, Stathmin was expressed in the cytoplasm and on the cell membranes of hDPSCs by cell immunofluorescence. Then, by constructing a lentiviral vector, the expression of Stathmin in hDPSCs was inhibited. Treatment with Stathmin shRNA (shRNA‐Stathmin group) inhibited the ability of hDPSCs to proliferate, as demonstrated by a CCK8 assay and flow cytometry analysis, and suppressed the osteogenic/odontoblastic differentiation ability, as demonstrated by alizarin red S staining and osteogenic/odontoblastic differentiation‐related gene (ALP, BSP, OCN, DSPP) activity, compared to that of hDPSCs from the control shRNA group. Molecular analyses showed that the Shh/GLI1 signalling pathway was inhibited when Stathmin was silenced, and purmorphamine, the Shh signalling pathway activator, was added to hDPSCs in the shRNA‐Stathmin group, real‐time PCR and Western blotting confirmed that expression of Shh and its downstream signalling molecules PTCH1, SMO and GLI1 increased significantly. After activating the Shh signalling pathway, the proliferation of hDPSCs increased markedly, as demonstrated by a CCK8 assay and flow cytometry analysis; osteogenic/odontoblastic differentiation‐related gene (ALP, BSP, OCN, DSPP) expression also increased significantly. Collectively, these findings firstly revealed that Stathmin‐Shh/GLI1 signalling pathway plays a positive role in hDPSC proliferation and osteogenic/odontoblastic differentiation.
Human umbilical cord mesenchymal stem cells facilitate the up‐regulation of miR‐153‐3p, whereby attenuating MGO‐induced peritoneal fibrosis in rats
Authors: Li Dong , Lu Zhenyu , Li Xiyuan , Xu Zhongwei , Jiang Jianqing , Zheng Zhenfeng , Jia Junya , Lin Shan , Yan Tiekun
Published: April 2018

MiRNAs contribute greatly to epithelial to mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs), which is a crucial step in peritoneal fibrosis (PF). In this study, we tried to profile whether miRNA expression differences exist after human umbilical cord mesenchymal stem cells (hUCMSCs) treatment in PF rats and investigate the possible role of miR‐153‐3p involved in anti‐EMT process. We randomly assigned 34 rats into three groups: control group (Group Control), MGO‐induced PF rats (Group MGO) and hUCMSCs‐treated rats (Group MGO + hUCMSCs). MiRNA microarrays and real‐time PCR analyses were conducted in three groups. α‐SMA, Snail1 and E‐cadherin expression were detected by Western blot. Luciferase reporter assays were used to detect the effects of miR‐153‐3p overexpression on Snai1 in rat peritoneal mesothelial cells (RPMCs). We identified differentially expressed miRNAs related to EMT, in which miR‐153‐3p demonstrated the greatest increase in Group MGO + hUCMSCs. Transient cotransfection of miR‐153‐3p mimics with luciferase expression plasmids resulted in a significant repression of Snai1 3′‐untranslated region luciferase activity in RPMCs. These studies suggest that miR‐153‐3p is a critical molecule in anti‐EMT effects of hUCMSCs in MGO‐induced PF rats. MiR‐153‐3p might exert its beneficial effect through directly targeting Snai1.
TP53INP1 inhibits hypoxia‐induced vasculogenic mimicry formation via the ROS/snail signalling axis in breast cancer
Authors: Wang Yi , Sun Huizhi , Zhang Danfang , Fan Dan , Zhang Yanhui , Dong Xueyi , Liu Shiqi , Yang Zhao , Ni Chunsheng , Li Yanlei , Liu Fang , Zhao Xiulan
Published: April 2018

Tumour protein p53‐inducible nuclear protein 1 (TP53INP1) is a tumour suppressor associated with malignant tumour metastasis. Vasculogenic mimicry (VM) is a new tumour vascular supply pattern that significantly influences tumour metastasis and contributes to a poor prognosis. However, the molecular mechanism of the relationship between TP53INP1 and breast cancer VM formation is unknown. Here, we explored the underlying mechanism by which TP53INP1 regulates VM formation in vitro and in vivo. High TP53INP1 expression was not only negatively correlated with a poor prognosis but also had a negative relationship with VE‐cadherin, HIF‐1α and Snail expression. TP53INP1 overexpression inhibited breast cancer invasion, migration, epithelial‐mesenchymal transition (EMT) and VM formation; conversely, TP53INP1 down‐regulation promoted these processes in vitro by functional experiments and Western blot analysis. We established a hypoxia model induced by CoCl2 and assessed the effects of TP53INP1 on hypoxia‐induced EMT and VM formation. In addition, we confirmed that a reactive oxygen species (ROS)‐mediated signalling pathway participated in TP53INP1‐mediated VM formation. Together, our results show that TP53INP1 inhibits hypoxia‐induced EMT and VM formation via the ROS/GSK‐3β/Snail pathway in breast cancer, which offers new insights into breast cancer clinical therapy.
Referendums in the European Union: Defective by Birth?
Authors: Cheneval Francis , Ferrín Mónica
Published: April 2018

On the basis of a combined examination of normative claims and empirical evidence this paper discusses minimal criteria for the institutional design of referendums on EU‐internal issues. These criteria concern the mandatory (vs. optional), the simultaneous (vs. serial) and binding (vs. consultative) nature of referendums. The proposed criteria are demanding, both for the Member States and the European Union, but experiences show that the EU is in fact participating actively in EU‐issues referendums and Member States as well as the EU need to surpass the current arbitrary use of plebiscites by governments. On a broader scale the paper contributes to the insight that it might be time to fully address the use of direct democracy at the national and EU levels.
Screening of candidate biomaterials for alveolar augmentation using a critical‐size rat calvaria defect model
Authors: Susin Cristiano , Lee Jaebum , Fiorini Tiago , Koo Ki‐Tae , Schüpbach Peter , Angst Patricia D. M. , Finger Stadler Amanda , Wikesjö Ulf ME
Published: April 2018

Objective

To screen candidate biomaterials intended for alveolar augmentation relative to their potential to enhance local bone formation using a routine critical‐size (ø8‐mm) rat calvaria defect model.

Methods

One hundred and forty male Sprague Dawley outbred rats, age 11–12 weeks, weight 325–375 g, obtained from USDA approved breeder, randomised into 14 groups of 10 animals, each received one of the following treatments: sham‐surgery (empty control), Bio‐Oss (bovine HA/reference control), or candidate biomaterials including bovine HA, synthetic HA/ß‐TCP and calcium phosphate constructs, mineralised/demineralised human bone preparations, a ß‐TCP/calcium sulphate and an HA/calcium sulphate putty. A 4‐week healing interval was chosen to discern local bone formation using incandescent and polarised light microscopy. Statistical analysis used one‐way ANOVA followed by Bonferroni for pairwise comparisons.

Results

Candidate biomaterials all displayed biocompatibility. They exhibited limited, if any, appreciable bioerosion or biodegradation. No statistically significant differences in mean linear defect closure were observed among experimental groups, sham‐surgery displaying the highest score (48.1 ± 24.3%). Sham‐surgery also showed a significantly greater bone area fraction than all other groups (19.8 ± 13.9%, p < .001). The HA/calcium sulphate putty showed a significantly greater residual biomaterial area fraction than all other groups (61.1 ± 8.5%, p < .01).

Conclusion

Within the limitations of this animal model, although biocompatible, none of the tested biomaterials enhanced local bone formation beyond the innate regenerative potential of this craniotomy defect.
Evaluation of the Relationship Between the 18S rRNA/rDNA Ratio and Population Growth in the Marine Diatom Skeletonema tropicum via the Application of an Exogenous Nucleic Acid Standard
Authors: Lin Yun‐Chi , Kang Lee‐Kuo , Shih Chi‐Yu , Gong Gwo‐Ching , Chang Jeng
Published: April 2018

Ribosomal RNA (rRNA) has been regarded as a proxy for metabolic activity and population growth in microbes, but the limitations and assumptions of this approach should be better defined, particularly in eukaryotic microalgae. In this study, the 18S rRNA/rDNA ratio of a marine diatom, Skeletonema tropicum, was examined in batch and semi‐continuous cultures subjected to low nitrogen and phosphorus treatments at a temperature of 20 °C. In the semi‐continuous cultures, the measured 18S rRNA/rDNA ratio ranged from 4.0 × 102 to 5.0 × 103, and the logarithmic form of this ratio increased linearly with the population growth rate under both low nitrogen and low phosphorus conditions. In batch cultures grown under low nitrogen or low phosphorus conditions, log (rRNA/rDNA) also increased linearly with growth rate when the latter ranged between −0.4 and 1.5 day−1. The 18S rRNA/rDNA ratios of Skeletonema sampled from in the southern East China Sea were substantially lower than measured from laboratory cultures. Among the field samples, ratios obtained at a coastal station were higher than those obtained farther offshore. These results imply higher growth rate at the coastal station, but the influences of other factors, such as cell size and temperature, cannot be ruled out.
An In Vitro Model of the Blood–Brain Barrier: Naegleria fowleri Affects the Tight Junction Proteins and Activates the Microvascular Endothelial Cells
Authors: Coronado‐Velázquez Daniel , Betanzos Abigail , Serrano‐Luna Jesús , Shibayama Mineko
Published: April 2018

Naegleria fowleri causes a fatal disease known as primary amoebic meningoencephalitis. This condition is characterized by an acute inflammation that originates from the free passage of peripheral blood cells to the central nervous system through the alteration of the blood–brain barrier. In this work, we established models of the infection in rats and in a primary culture of endothelial cells from rat brains with the aim of evaluating the activation and the alterations of these cells by N. fowleri. We proved that the rat develops the infection similar to the mouse model. We also found that amoebic cysteine proteases produced by the trophozoites and the conditioned medium induced cytopathic effect in the endothelial cells. In addition, N. fowleri can decrease the transendothelial electrical resistance by triggering the destabilization of the tight junction proteins claudin‐5, occludin, and ZO‐1 in a time‐dependent manner. Furthermore, N. fowleri induced the expression of VCAM‐1 and ICAM‐1 and the production of IL‐8, IL‐1β, TNF‐α, and IL‐6 as well as nitric oxide. We conclude that N. fowleri damaged the blood–brain barrier model by disrupting the intercellular junctions and induced the presence of inflammatory mediators by allowing the access of inflammatory cells to the olfactory bulbs.
Mitochondrial Genome of Vannella croatica (Amoebozoa, Discosea, Vannellida)
Authors: Bondarenko Natalya I. , Nassonova Elena S. , Mijanovic Olja , Glotova Anna A. , Kamyshatskaya Oksana G. , Kudryavtsev Alexander A. , Masharsky Alexey E. , Polev Dmitrii E. , Smirnov Alexey V.
Published: April 2018

Mitochondrial genome sequence of Vannella croatica (Amoebozoa, Discosea, Vannellida) was obtained using pulse‐field gel electrophoretic isolation of the circular mitochondrial DNA, followed by the next‐generation sequencing. The mitochondrial DNA of this species has the length of 28,933 bp and contains 12 protein‐coding genes, two ribosomal RNAs, and 16 transfer RNAs. Vannella croatica mitochondrial genome is relatively short compared to other known amoebozoan mitochondrial genomes but is rather gene‐rich and contains significant number of open reading frames.
Spirit‐Filled Geopolitics: Pentecostal Ontologies and the Honduran Coup
Authors: Girard William M.
Published: April 2018

Situado en Copán Ruinas, Honduras, el trabajo de campo en que está basado este artículo, describe el papel de la ontología Cristiana Pentecostal en las lógicas que sustentaron apoyo generalizado de los cristianos al golpe de estado que derrocó al presidente, Manuel Zelaya, en 2009. La literatura reciente que describe cómo el compromiso de algunos movimientos indígenas latinoamericanos representa un quiebre con los marcos modernos de la “política” en aras de argumentar que la forma de hacer política de los pentecostales también es no‐moderna. Entre los elementos no‐modernos de la política pentecostal —a la cual me refiero como “Geopolítica del Espíritu”— está tanto una temporalidad apocalíptica como una noción del campo de política que incorpora “lo sobrenatural”. El artículo utiliza la literatura sobre movimientos indígenas como un marco para describir las formas en que esta política pentecostal se entrecruza, sin reducirse, con el neoliberalismo y con las prácticas e imaginarios del estado‐nación laico, en particular en lo referido a la geopolítica de la Guerra Fría en los ochenta. [Honduras, Centroamérica, cristianismo, geografía, Guerra Fría, ontología]
Sparse‐coding denoising applied to reversible conformational switching of a porphyrin self‐assembled monolayer induced by scanning tunnelling microscopy
Authors: OLIVEIRA J. , BRAGANÇA A.M. , ALCÁCER L. , MORGADO J. , FIGUEIREDO M. , BIOUCAS‐DIAS J. , FERREIRA Q.
Published: April 2018

Scanning tunnelling microscopy (STM) was used to induce conformational molecular switching on a self‐assembled monolayer of zinc‐octaethylporphyrin on a graphite/tetradecane interface at room temperature. A reversible conformational change controlled by applying a tip voltage was observed. Consecutive STM images acquired at alternating tip voltages showed that at 0.4 V the porphyrin monolayer presents a molecular arrangement formed by alternate rows with two different types of structural conformations and when the potential is increased to 0.7 V the monolayer presents only one type of conformation. In this paper, we characterize these porphyrin conformational dynamics by analyzing the STM images, which were improved for better quality and interpretation by means of a denoising algorithm, adapted to process STM images from state of the art image processing and analysis methods.

STM remains the best technique to ‘see’ and to manipulate the matter at atomic scale. A very sharp tip a few angstroms of the surface can provide images of molecules and atoms with a powerful resolution. However, these images are strongly affected by noise which is necessary to correct and eliminate. This paper is about new computational tools specifically developed to denoise the images acquired with STM. The new algorithms were tested in STM images, obtained at room temperature, of porphyrin monolayer which presents reversible conformational change in function of the tip bias voltage. Images with high resolution, acquired in real time, show that the porphyrins have different molecular arrangements whether the tip voltage is 0.4 V or 0.7 V.
Hemodynamic Changes May Indicate Vessel Wall Injury After Stent Retrieval Thrombectomy for Acute Stroke
Authors: Perren Fabienne , Kargiotis Odysseas , Pignat Jean‐Michel , Pereira Vitor Mendes
Published: April 2018

BACKGROUND AND PURPOSE

Stent retrievers have revolutionized endovascular treatment of acute ischemic stroke (AIS). Animal studies showed that mechanical thrombectomy (MT) may cause endothelial injury and intimal layer edema. Using transcranial color‐coded duplex‐sonography (TCCS) we observed postprocedural hemodynamic changes in the treated vessel.

METHODS

We studied AIS patients with large intracranial artery occlusion in whom MT with stent retrievers was performed. Only those with complete recanalization (modified TICI‐2b or 3) as assessed by postprocedural digital subtraction angiography (DSA) and in whom early control TCCS was performed were retained. Patients treated with intra‐arterial thrombolysis or stenting were excluded.

RESULTS

In 31 patients treated within a time period of 4 years (29 with middle cerebral artery [MCA] and 2 with basilar artery [BA] occlusion), postacute stroke brain‐DSA confirmed complete recanalization without residual stenosis or vasospasm. However, in 27 (17 men, mean age 66.3 years) of them TCCS (mean 3.4 days after MT) showed very segmental acceleration of blood flow velocities in the affected arteries (MCA maximum peak systolic velocity [PSVmax] at least >35% as compared to the contralateral side at the same depth; BA PSVmax >40% as compared to velocities at different depths of the same vessel). None showed clinical deterioration. TCCS follow‐up (mean 20 days) showed normalization in 14 of 16 cases.

CONCLUSION

Our TCCS study provides preliminary evidence of focal acceleration of blood flow velocities after MT. Without residual stenosis or vasospasm, this may be a sign of endothelial layer disruption/intimal injury. Further studies are needed to confirm our results.
An In Vitro Evaluation of the Effect of Various Adhesives and Surface Treatments on Bond Strength of Resin Cement to Polyetheretherketone
Authors: Caglar Ipek , Ates Sabit Melih , Yesil Duymus Zeynep
Published: April 2018

Purpose

To investigate the effect of various surface pretreatments and adhesive systems on bond strength of resin cement to polyetheretherketone (PEEK).

Materials and Methods

360 PEEK specimens were divided into 4 groups to receive the following pretreatments (n = 90): no pretreatment, sandblasting, silica coating and Er:YAG laser. Surface roughness (SR) and scanning electron microscopy (SEM) evaluations were conducted after mechanical pretreatments for topographical surface evaluations. After the pretreatments, each group was divided into 3 subgroups (n = 30) according to the adhesive system used: no adhesive system, Visio.link, and Signum PEEK Bond. Resin cement was applied, and specimens underwent 5000 thermocycles (5‐55°C) before shear bond strength (SBS) test. One‐way ANOVA and Tukey test were used to analyze the SR data; 2‐way ANOVA and Fisher's LSD test were used to analyze the SBS data at the confidence interval of 95% (α = 0.05).

Results

The sandblasting group demonstrated the highest SR values (p < 0.05). No significant differences in SR values were identified among silica coating, laser, and control groups (p ˃ 0.05). Conditioning with Visio.link after sandblasting group exhibited the highest SBS values (p < 0.05). No significant differences in SBS values were found between laser and control group (p ˃ 0.05). Conditioned groups presented higher SBS values than unconditioned groups (p < 0.05). Between the two adhesive systems, Visio.link demonstrated statistically significant higher SBS values than Signum PEEK Bond (p < 0.05).

Conclusion

Silica coating and sandblasting were shown to provide superior pretreatment of PEEK surface. Laser pretreatments showed no effectiveness on bond strength of resin cement to PEEK. Adhesive systems improved the resin bonding to PEEK.
Management of fever and neutropenia in children with cancer: A survey of Australian and New Zealand practice
Authors: Haeusler Gabrielle M , Slavin Monica A , Bryant Penelope A , Babl Franz E , Mechinaud Francoise , Thursky Karin A
Published: April 2018

Aim

Variation in the management of fever and neutropenia (FN) in children is well described. The aim of this study was to explore the current management of FN across Australia and New Zealand and highlight areas for improvement.

Methods

A practice survey was administered to paediatric health‐care providers via four clinical and research networks. Using three clinical case vignettes, we explored risk stratification, empiric antibiotics, initial investigations, intravenous–oral switch, ambulatory management and antibiotic duration in children with cancer and FN.

Results

A response was received from 104 participants from 16 different hospitals. FN guideline compliance was rated as moderate or poor by 24% of respondents, and seven different fever definitions were described. There was little variation in the selected empiric monotherapy and dual‐therapy regimens, and almost all respondents recommended first‐dose antibiotics within 1 h. However, 27 different empiric antibiotic combinations were selected for beta‐lactam allergy. An incorrect risk status was assigned to the low‐risk case by 27% of respondents and to the high‐risk case by 41%. Compared to current practice, significantly more respondents would manage the low‐risk case in the ambulatory setting provided adequate resources were in place (43 vs. 85%, P < 0.0001). There was variation in the use of empiric glycopeptides as well as use of aminoglycosides beyond 48 h.

Conclusion

Although the antibiotics selected for empiric management of FN are appropriate and consistent, variation and inaccuracies exist in risk stratification, the selection of monotherapy over dual therapy, empiric antibiotics chosen for beta‐lactam allergy, use of glycopeptides and duration of aminoglycosides.
Efficacy and safety of human chorionic gonadotropin for treatment of cryptorchidism: A meta‐analysis of randomised controlled trials
Authors: Wei Yi , Wang Yangcai , Tang Xiangliang , Liu Bin , Shen Lanju , Long Chunlan , Lin Tao , He Dawei , Wu Shengde , Wei Guanghui
Published: April 2018

Aim

Although human chorionic gonadotropin (hCG) has long been employed in the management of cryptorchidism, its safety and efficacy is still controversial. Hence, in the present study, we conducted a meta‐analysis of the treatment of cryptorchidism using hCG.

Methods

We searched the Medline, Embase, CINAHL, EBSCO, The Cochrane Library, China National Knowledge Infrastructure and WanFang databases. Data were extracted by two reviewers using the designed extraction form. Data up to July 2015 were obtained using the terms ‘cryptorchidism’, ‘chorionic gonadotropin’ and ‘randomised controlled trials’. All the publications were downloaded, and the respective authors were contacted for any further details and clarifications, if deemed necessary. The data analysis included randomised controlled trials that compared hCG with other hormone treatments offered to prepubescent males presenting with cryptorchidism. Testicular descent rate was used as the final positive outcome of the treatments offered. The software Review Manager (RevMan 5.3, The Cochrane Collaboration, London, UK) was used to review the management and data analysis. Risk ratios (RRs) with 95% confidence intervals (CIs) were pooled with a fixed effect model if no heterogeneity was present.

Results

A total of seven trials satisfied the selection criteria. The overall quality of the studies downloaded from various databases was low. Data from these seven studies were divided into three subgroups depending on the design of the trials: Two studies compared hCG with a placebo, and three studies compared hCG with gonadotropin‐releasing hormone (GnRH) in unilateral cryptorchidism, whereas two other studies compared hCG with GnRH in bilateral cryptorchidism. Analysis of these trials revealed no significant differences between the effectiveness of hCG treatment and GnRH treatment in bilateral (RR 0.05, 95% CI (−0.29–0.40), two trials, n = 104, P = 0.76) as well as unilateral cryptorchidism (RR 0.04, 95% CI (−0.12, 0.21), three trials, n = 81, P = 0.61). A meta‐analysis of these studies showed that hCG treatment is not superior to placebo (RR 7.74, 95% CI (0.14–425.72), two trials, n = 31, P = 0.32).

Conclusion

A meta‐analysis of the seven studies led us to conclude that hCG treatment is no more effective than placebo, and there were no significant differences in the effectiveness of hCG versus GnRH treatment.
Melatonin disturbs SUMOylation‐mediated crosstalk between c‐Myc and nestin via MT1 activation and promotes the sensitivity of paclitaxel in brain cancer stem cells
Authors: Lee Hyemin , Lee Hyo‐Jung , Jung Ji Hoon , Shin Eun Ah , Kim Sung‐Hoon
Published: April 2018

Here the underlying antitumor mechanism of melatonin and its potency as a sensitizer of paclitaxel was investigated in X02 cancer stem cells. Melatonin suppressed sphere formation and induced G2/M arrest in X02 cells expressing nestin, CD133, CXCR4, and SOX‐2 as biomarkers of stemness. Furthermore, melatonin reduced the expression of CDK2, CDK4, cyclin D1, cyclin E, and c‐Myc and upregulated cyclin B1 in X02 cells. Notably, genes of c‐Myc related mRNAs were differentially expressed in melatonin‐treated X02 cells by microarray analysis. Consistently, melatonin reduced the expression of c‐Myc at mRNA and protein levels, which was blocked by MG132. Of note, overexpression of c‐Myc increased the expression of nestin, while overexpression of nestin enhanced c‐Myc through crosstalk despite different locations, nucleus, and cytoplasm. Interestingly, melatonin attenuated small ubiquitin‐related modifier‐1 (SUMO‐1) more than SUMO‐2 or SUMO‐3 and disturbed nuclear translocation of nestin for direct binding to c‐Myc by SUMOylation of SUMO‐1 protein by immunofluorescence and immunoprecipitation. Also, melatonin reduced trimethylated histone H3K4me3 and H3K36me3 more than dimethylation in X02 cells by Western blotting and chromatin immunoprecipitation assay. Notably, melatonin upregulated MT1, not MT2, in X02 cells and melatonin receptor inhibitor luzindole blocked the ability of melatonin to decrease the expression of nestin, p‐c‐Myc(S62), and c‐Myc. Furthermore, melatonin promoted cytotoxicity, sub‐G1 accumulation, and apoptotic body formation by Paclitaxcel in X02 cells. Taken together, these findings suggest that melatonin inhibits stemness via suppression of c‐Myc, nestin, and histone methylation via MT1 activation and promotes anticancer effect of Paclitaxcel in brain cancer stem cells.
Melatonin protects prepuberal testis from deleterious effects of bisphenol A or diethylhexyl phthalate by preserving H3K9 methylation
Authors: Zhang Teng , Zhou Yang , Li Lan , Zhao Yong , De Felici Massimo , Reiter Russel J. , Shen Wei
Published: April 2018

A growing number of couples experience fertility issues with almost half being due to malefactors. The exposure to toxic environmental contaminants, such as endocrine disruptors (EDs), has been shown to negatively affect male fertility.EDs are present in the environment, and exposure to these toxins results in the failure of spermatogenesis. The deleterious effects ofEDs on spermatogenesis have been well documented, whereas improvement of infertility associated with spermatogenesis defects remains a great challenge. Herein, we report that in vitro exposure of prepuberal mouse testes to two well‐known endocrine disruptors (EDs), bisphenol A (BPA) or diethylhexyl phthalate (DEHP), impairs spermatogenesis with perturbing self‐renewal, spermatogonia activity, and meiosis. Evidence indicates that such effects are likely due, at least in part, to decreased G9a‐dependent H3K9 di‐methylation. Of note, we found that melatonin (MLT) protected the testis from the negativeEDimpacts with preserving spermatogonia stem and meiotic cells, along with maintaining normal H3K9 di‐methylation in these cells. Taken together, this work documents thatBPAandEDHPadversely affect prepuberal spermatogenesis and perturb crucial epigenetic activities in male germ cells and highlight the protective ability ofMLT.
Melatonin promotes sleep in mice by inhibiting orexin neurons in the perifornical lateral hypothalamus
Authors: Sharma Rishi , Sahota Pradeep , Thakkar Mahesh M.
Published: April 2018

Melatonin promotes sleep. However, the underlying mechanisms are unknown. Orexin neurons in the perifornical lateral hypothalamus (PFH) are pivotal for wake promotion. Does melatonin promote sleep by inhibiting orexin neurons? We used C57BL/6J mice and designed 4 experiments to address this question. Experiment 1 used double‐labeled immunofluorescence and examined the presence of melatonin receptors on orexin neurons. Second, mice, implanted with bilateral guides targeted toward PFH and sleep‐recording electrodes, were infused with melatonin (500 pmole/50 nL/side) at dark onset (onset of active period), and spontaneous bouts of sleep‐wakefulness were examined. Third, mice, implanted with bilateral guides into the PFH, were infused with melatonin (500 pmole/50 nL/side) at dark onset and euthanized 2 hours later, to examine the activation of orexin neurons using c‐Fos expression in orexin neurons. Fourth, mice, implanted with PFH bilateral guides and sleep‐recording electrodes, were infused with melatonin receptor antagonist, luzindole (10 pmol/50 nL/side), at light onset (onset of sleep period), and spontaneous bouts of sleep‐wakefulness were examined. Our results suggest that orexin neurons express MT1, but not MT2 receptors. Melatonin infusion into the PFH, at dark onset, site‐specifically and significantly increased NREM sleep (43.7%, P = .003) and reduced wakefulness (12.3%, P = .013). Local melatonin infusion at dark onset inhibited orexin neurons as evident by a significant reduction (66%, P = .0004) in the number of orexin neurons expressing c‐Fos. Finally, luzindole infusion‐induced blockade of melatonin receptors in PFH at sleep onset significantly increased wakefulness (44.1%, P = .015). Based on these results, we suggest that melatonin may act via the MT1 receptors to inhibit orexin neurons and promote sleep.
Digestive proteases and in vitro protein digestibility of feed ingredients for the Central American river turtle, Dermatemys mawii
Authors: Rangel‐Mendoza J. , Hernández‐García J. , Álvarez‐González C. A. , Guerrero‐Zárate R. , Zenteno‐Ruiz C. E. , López‐Luna M. A.
Published: April 2018

Functional characteristics of digestive proteases and in vitro digestibility of several protein sources were studied in hatchlings of Central American river turtles, Dermatemys mawii. Acidic and alkaline proteases from the stomach and intestines were used, and optimums for acidic proteases were registered at 55°C and pH 2, while alkaline proteases were found at 55°C and pH 9. Ten protein ingredients, both vegetable and animal sources, were explored by the pH‐STAT method, only for alkaline proteases. The degree of hydrolysis was at its highest for squid meal and lowest for blood meal, while the total free amino acids level was at its highest for squid meal, and lowest for wheat gluten meal. Our results indicate that D. mawii has a broad capacity to digest both animal and vegetable sources, and suggests some ingredients more suitable to design artificial diets for this species.
In vitro gas and methane production in rumen fluid from dairy cows fed grass silages differing in plant maturity, compared to in vivo data
Authors: Macome F. M. , Pellikaan W. F. , Hendriks W. H. , Warner D. , Schonewille J. T. , Cone J. W.
Published: April 2018

The relationship between in vitro rumen CH4 production of grass silages, using the gas production technique, and in vivo data obtained with the same cows and rations in respiration chambers was investigated. Silages were made from grass harvested in 2013 on May 6th, May 25th, July 1st and July 8th. The grass silages were used to formulate four different rations which were fed to 24 cows in early and late lactation, resulting in a slightly different dry matter intake (DMI; 16.5 kg/day vs. 15.4 kg/day). The experimental rations consisted of 70% grass silage, 10% maize silage, and 20% concentrates on a dry matter basis. Cows were adapted to the rations for 17 days before rumen fluid was collected via oesophageal tubing, and in vitro gas and CH4 production were analysed. In vitro total gas and CH4 production of the (ensiled) grass expressed as ml/g OM decreased with advancing maturity of the grass. The in vitro CH4 production after 48 hr of incubation expressed in ml/g OM did not correlate with the in vivo CH4 production expressed in g/kg organic matter intake or g/kg DMI (R2 = .00–.18, p ≥ .287). The differences in CH4 emission per unit of intake observed in vivo were rather small between the different rations, which also contributed to the observed poor relationship. Utilizing stepwise multiple regression improved the correlation only slightly. In vitro gas and CH4 production varied based on whether donor cows were previously adapted to the respective ration or not, suggesting that careful adaption to the experimental diet should be envisaged in in vitro gas and CH4 production experiments.
Investigation of carryover effect of prior fibre consumption on growth, serum and tissue metabolic markers in Ossabaw pigs fed a high‐fat diet
Authors: Almeida V. V. , Yan H. , Nakatsu C. H. , Ajuwon K. M.
Published: April 2018

Carryover effect of prior fibre consumption on metabolic markers was investigated. Treatments were arranged in 2 × 2 factorial with 2 fibre sources, 4% inulin or cellulose (Solka‐Floc®) and fat levels (5 or 15%) for the low‐fat diet (LFD) and high‐fat diet (HFD) respectively. Pigs were fed the two fibre diets for the first 56d (nursery phase), and thereafter fed either the LFD or HFD containing no added fibre source from d56 to 140 (growing phase). Pigs on the HFD were heavier (p = .05) than those on LF (64.61 vs. 68.38 kg), regardless of prior fibre type consumed. Pigs that were fed cellulose during the nursery and later fed the HFD had the highest ADG (p < .05). Feeding the HFD resulted in higher back fat (BF) (13.41 and 18.18 ± 0.12 mm for LFD and HFD, respectively; p < .01). The HFD resulted in higher (p < .01) insulin (0.014 and 0.016 ± 0.001 mg/L for LF and HF respectively) and glucose (100.89 and 125.03 ± 4.39 mg/dl for LF and HF respectively) concentrations in the serum. Inulin increased (p ≤ .02) jejunal expression of SREBP‐1c and CL‐4, but reduced (p < .05) TNFɑ and IL‐6 expression in the ileum. Alpha‐diversity was significantly different (p < .05) between the inulin and cellulose fed pigs at the end of the nursery and finishing phases. Therefore, inulin feeding before a HFD may lead to reduction in ADG and inflammatory markers in the small intestine of pigs, and thus prevent future metabolic disorders.
The antimicrobial activity of lavender essential oil (Lavandula angustifolia) and its influence on the production performance of broiler chickens
Authors: Adaszyńska‐Skwirzyńska M. , Szczerbińska D.
Published: April 2018

The aim of the study was the evaluation of the antimicrobial activity (in vitro) of lavender (Lavandula angustifolia) essential oil (LEO) and the effect of its addition to the drinking water of broiler chickens on their production performance. Antimicrobial activity was determined by establishing the minimum inhibitory concentration (MIC) using a series of microdilutions. Bird experiments were carried out on a commercial farm on 300 Ross 308 broilers. One‐day‐old chicks were randomly assigned to three experimental groups of 100 individuals (five replications of 20 individuals each). In the control group, chickens received drinking water without added essential oil throughout the rearing period. In the LEO0.2 and LEO0.4 groups, from 1 to 42 days of bird life, the LEO0.2 group had 0.2 ml/L of essential lavender oil added to the drinking water, while LEO0.4 had 0.4 ml/L added. The results of the experiment showed the antimicrobial activity of LEO and its positive effect on the production results of broiler chickens. Application of higher concentration of essential oil (0.4 ml/L) significantly affected production results (BW, FCR, WCR—p < .01). No differences were observed in FI, WI, F:W and mortality (p > .05). In vitro studies indicate a significant effect of LEO on the inhibition of microbial growth. These results encourage further studies on a larger scale that will confirm antimicrobial efficiency and define the mechanisms of action of Lavandula angustifolia essential oil and its individual components.
Inhibition of Internal Attention to Positive and Negative Representations in Dysphoria
Authors: Isato Ayako , Nishimura Haruki , Yamada Makiko , Mochizuki Satoshi
Published: April 2018

We developed the Internal–External Attention‐Switching Task, which measures the ability to inhibit internal attention and the ability to remove inhibition of internal attention. The Internal–External Attention‐Switching Task was administered to a sample of dysphoric and non‐dysphoric undergraduate or graduate students (N = 44). Difficulty in both inhibition of internal attention to negative representation and removal of this inhibition were shown to be general. Importantly, however, dysphoric individuals could not maintain internal attention or remove inhibition of internal attention to positive representations as easily as non‐dysphoric individuals could. Therefore, reduced attention to positive representations in dysphoric individuals may enable maintenance of their depressive mood.
Emerging psychiatric disorders in Kleine‐Levin syndrome
Authors: Groos Elisabeth , Chaumereuil Charlotte , Flamand Mathilde , Brion Agnes , Bourdin Hubert , Slimani Vanessa , Lecendreux Michel , Arnulf Isabelle
Published: April 2018

In Kleine‐Levin syndrome (KLS), episodes of hypersomnia and cognitive, psychiatric and behavioural disturbances alternate with asymptomatic periods in adolescents. We evaluated whether psychiatric disorders would emerge during asymptomatic periods in a naturalistic, uncontrolled clinical cohort. Patients with primary KLS underwent psychiatric interviews at diagnosis and every year for 1–10 years, leading to diagnosis of former and present comorbid psychiatric disorders. Among the 115 patients (65.2% male and aged 16.1 ± 4.8 years at KLS onset), 19 (16.5%) had a history of psychiatric disorder prior to KLS onset, which persisted afterwards in 10. Twenty‐five (21%) patients developed a new, comorbid psychiatric disorder 1–6 years after KLS onset, during ‘asymptomatic’ periods, including mood disorders (n = 14; including major depressive episodes, n = 8; recurrent depressive episodes, n = 2; bipolar I disorder, n = 1; dysthymic disorder, n = 1; adjustment disorder with depressive mood, n = 1; and mood disorder not otherwise specified, n = 1), anxiety disorders (n = 7), eating disorders (n = 2), psychotic disorders not otherwise specified (n = 2), schizoaffective disorder (n = 1) and cannabis dependence (n = 1). Six patients attempted suicide: two before and two after KLS onset, and two during episodes. Female sex, longer disease course, longer time incapacitated (356 ± 223 versus 155 ± 186 days) and more frequent psychiatric symptoms during episodes (but no family or personal history of psychiatric disorders) were associated with emerging psychiatric disorders. Contrary to the alleged benignity of KLS and normality between episodes, one KLS patient in five suffers from emerging psychiatric disorders. These disorders may depend on personal vulnerability and, most probably, on psychiatric symptoms during episodes.
Schizotypal traits are associated with sleep spindles and rapid eye movement in adolescence
Authors: Kuula Liisa , Merikanto Ilona , Makkonen Tommi , Halonen Risto , Lahti‐Pulkkinen Marius , Lahti Jari , Heinonen Kati , Räikkönen Katri , Pesonen Anu‐Katriina
Published: April 2018

Research suggests an association between schizophrenia and a decrease in sleep spindle activity, as well as a change in sleep architecture. It is unknown how the continuum of psychotic symptoms relates to different features in the sleep electroencephalogram. We set out to examine how sleep architecture and stage 2 spindle activity are associated with schizotypy in a healthy adolescent population. The participants in our study (n = 176, 61% girls) came from a community‐based cohort. Schizotypal traits were evaluated using the Schizotypal Personality Scale (STA) in early adolescence (mean age 12.3 years, SD = 0.5) and the participants underwent ambulatory overnight polysomnography at mean age 16.9 years (SD = 0.1). Sleep was scored in 30‐s epochs into stages 1, 2, 3 and rapid eye movement (REM) sleep. Stage 2 spindles were detected using an automated algorithm. Spindle analyses from central and frontal derivations included spindle duration and density for slow (10–13 Hz) and fast (13–16 Hz) ranges. Covariates included sex and age. Those with the highest STA scores had a higher percentage of REM (B = 2.07 [95% CI, 0.17, 4.0]; p = .03) than those with the lowest scores. Those with the highest scores had shorter spindle duration, as derived from the frontal regions, and a slower oscillation range (B = −0.04 [95% CI, −0.07, −0.01]; p = .023) than those with the lowest scores. We conclude that high levels of schizotypy characteristics measured in early adolescence may be associated with distinguished features of sleep architecture, namely with spindle morphology and a higher proportion of REM sleep.
Effects of enrofloxacin on antioxidant system, microsomal enzymatic activity, and proteomics in porcine liver
Authors: Li Y. , Mou Y. , Thunders M. , Wu Y. , Ai X. , Zhou X. , Qiu J.
Published: April 2018

Enrofloxacin (EF) is a widely used fluoroquinolone, usually regarded as a safe and effective treatment for bacterial infections. Adverse effects of EF have previously been demonstrated in some species, but so far there have been no studies looking specifically at the impact of EF on pigs. In this study, three different doses of EF (5, 25 and 125 mg kg bw−1) were administrated to Bama pigs. The results showed that lipid peroxidation of pig liver tissue occurred with all EF doses. The 125 mg kg dose of EF induced catalase (CAT) and glutathione peroxidase (GSH‐px) and increased CYP450 content in pig liver microsomes. The activity of microsomal NADPH‐cytochrome C reductase (NCCR) was elevated at both the 25 and 125 mg kg doses of EF. Microsomal erythromycin N‐demethylase (ERND) and aminopyrin N‐demethylase (AND) were inhibited by high doses of EF, while aniline‐4‐hydroxylase (AH) was unaffected. None of the EF treatments affected superoxide dismutase (SOD) or cytochrome b5 content. Antioxidases and microsomal enzymes may work together to resist the adverse effects of EF. Proteomic analysis revealed increased protein expression of carboxylesterase (CES) and alpha‐enolase (ENO1) in microsomes as a stress response to EF. These results provide new information about the adverse effect of fluoroquinolones and help guide their usage more effectively in the clinic or animal breeding.
Pharmacokinetics of cefquinome in crucian carp (Carassius auratus gibelio) after oral, intramuscular, intraperitoneal, and bath administration
Authors: Shan Q. , Wang J. , Yang F. , Ma L. , Yin Y. , Liu S. , Li L. , Zheng G.
Published: April 2018

The pharmacokinetics (PK) of cefquinome (CEQ) was studied in crucian carp (Carassius auratus gibelio) after single oral, intramuscular (i.m.), and intraperitoneal (i.p.) administration at a dose of 10 mg/kg body weight and following incubation in a 5 mg/L bath for 5 hr at 25°C. The plasma concentration of CEQ was determined using high‐performance liquid chromatography (HPLC). PK parameters were calculated based on mean CEQ concentration using WinNonlin 6.1 software. The disposition of CEQ following oral, i.m., or i.p. administration was best described by a two‐compartment open model with first‐order absorption. After oral, i.m., and i.p. administration, the maximum plasma concentration (Cmax) values were 1.52, 40.53, and 67.87 μg/ml obtained at 0.25, 0.23, and 0.35 hr, respectively, while the elimination half‐life (T1/2β) values were 4.68, 7.39, and 6.88 hr, respectively; the area under the concentration–time curve (AUC) values were 8.61, 339.11, and 495.06 μg hr/ml, respectively. No CEQ was detected in the plasma after bath incubation. Therapeutic blood concentrations of CEQ can be achieved in the crucian carp following i.m. and i.p. administration at a dosage of 10 mg/kg once every 2 days.
Development and validation of a novel score for fibrosis staging in patients with chronic hepatitis B
Authors: Wu Daxian , Rao Qunfang , Chen Wenqian , Ji Feiyang , Xie Zhongyang , Huang Kaizhou , Chen Er'mei , Zhao Yalei , Ouyang Xiaoxi , Zhang Sainan , Jiang Zhengyi , Zhang Lingjian , Xu Linjie , Gao Hainv , Li Lanjuan
Published: April 2018

Background & Aims

Non‐invasive assessment methods for liver fibrosis are urgently needed. The present study aimed to develop a novel diagnostic model for fibrosis staging in patients with chronic hepatitis B.

Methods

A cross‐sectional set of 417 chronic hepatitis B patients who underwent liver biopsy was enrolled and the METAVIR score was adopted as the reference of fibrosis staging.

Results

Among thyroid hormones, only the level of free tetraiodothyronine (FT4) decreased gradually with the METAVIR fibrosis score (P < .001). FibroStage, a novel diagnosis model that incorporates data on FT4, platelets, cholinesterase, gamma‐glutamyl transpeptidase, and age, was developed using the deriving set (n = 219). For the diagnosis of significant fibrosis, the FibroStage model had a significantly higher area under the receiver operating curve than did the FibroIndex, Forn, and Lok models (all of P < .01) and tended to better than the fibrosis‐4 (P = .0791) but comparable with the aspartate transaminase‐to‐platelet ratio index model (P = .1694). For the diagnosis of advanced fibrosis, FibroStage had a higher area under the receiver operating curve than did the aspartate transaminase‐to‐platelet ratio index, FibroIndex, Forn, and Lok models (all of P < .05) and had a comparable area under the receiver operating curve with the fibrosis‐4 model (P = .2109). For the diagnosis of cirrhosis, the area under the receiver operating curve of FibroStage was higher than those of the aspartate transaminase‐to‐platelet ratio index, fibrosis‐4, FibroIndex, and Lok (all of P < .05) models and was comparable with Forn (P = .1649). These results was validated by a validation set (n = 198).

Conclusion

FT4 may be an indicator for fibrosis staging in chronic hepatitis B patients. FibroStage is a better model than aspartate transaminase‐to‐platelet ratio index, fibrosis‐4, FibroIndex, Forn, and Lok for the comprehensively diagnosis of significant and advanced fibrosis and cirrhosis.
Sp1‐regulated transcription of RasGRP1 promotes hepatocellular carcinoma (HCC) proliferation
Authors: Zhang Xinran , Zhuang Hao , Han Feng , Shao Xiaowen , Liu Yun , Ma Xuda , Wang Zun , Qiang Zhaoyan , Li Yongmei
Published: April 2018

Background and Aims

The role of Ras guanine nucleotide‐releasing protein 1 (RasGRP1) in tumourigenesis has been a subject of debate, and its functions and clinical significance in hepatocellular carcinoma (HCC) remain unknown. Here, we evaluated the expression of RasGRP1 in HCC and determined how it contributes to HCC cell proliferation.

Methods

RasGRP1 expression was measured by quantitative polymerase chain reaction (qPCR) and Western blotting of 24 paired HCC tissues and para‐tumour tissues. RasGRP1 expression was confirmed by immunohistochemical analysis of a tissue microarray from 1 independent cohort. Overall survival (OS) and disease‐free survival (DFS) were estimated using the Kaplan‐Meier method, and risk factors that contributed to OS or DFS were identified using Cox regression analysis. The biologic relevance of RasGRP1 was examined by small interfering RNAs and an exogenous plasmid construct. Chromatin immunoprecipitation assays were performed to examine the binding of Sp1 to the RasGRP1 promoter.

Results

Increased RasGRP1 expression was associated with tumour size (P = .004), tumour‐node‐metastasis stage (P = .032), and Barcelona Clinic Liver Cancer stage (P = .002). RasGRP1 overexpression was an independent prognostic factor in HCC patients. RasGRP1 downregulation inhibited cell proliferation, whereas RasGRP1 overexpression promoted cell proliferation. Moreover, specificity protein 1 bound to the RasGRP1 promoter and promoted RasGRP1 transcription. In addition, RasGRP1 overexpression enhanced activation of the c‐Raf pathway.

Conclusions

RasGRP1 is upregulated in HCC and promotes HCC cell proliferation. Thus, RasGRP1 may be a novel therapeutic target for HCC.