bluuebunny/crossref_metadata_2025 huggingface dataset:
Why do German physicians reject managed care?
Authors: Ehlert Andree , Oberschachtsiek Dirk
Published: August 2018

Background

Approximately 20 years after the launch of managed care (MC) in Germany, the initial dynamics have turned into an MC backlash with a poor image for MC among physicians and the insured. Factors in MC contract rejection by physicians have not previously been studied systematically.

Objective

The objective of the study is to show that observed physician characteristics explain MC contract rejection in a quantitative model. These econometric findings will be related to suitable political measures to foster physicians' MC acceptance.

Methods

Results are based on 500 completed responses to a survey among practicing physicians in Northern Germany. Physician cooperation, attitudes toward MC, and practice characteristics are addressed. A Heckman approach accounts for potential preselection of physicians by insurers. Econometric findings are complemented by a qualitative analysis of free‐text answers.

Results

Private patient share, physician age, and number of physicians per practice significantly increase rejection probability, whereas it is decreased by rural location. Qualitatively, administrative burden and loss of professional autonomy are the main reasons for MC refusal.

Conclusion

Current health policy focuses on institutional measures such as innovation funding to promote MC. Our results show that it may be more effective to turn attention to practicing physicians' preferences as a bottleneck to MC development.
Moving towards universal health coverage: The need for a strengthened planning process
Authors: Nabyonga‐Orem Juliet , Nabukalu Belinda J. , Andemichael Ghirmay , Khosi‐Mthetwa Rosemary , Shaame Attiye , Myeni Sebentile , Quinto Ebony , Dovlo Delanyo
Published: August 2018

As countries embrace the ambitious universal health coverage (UHC) agenda whose major tenents include reaching everyone with the needed good quality services, strengthening the planning process to work towards a common objective is paramount. Drawing from country experiences—Swaziland and Zanzibar, we reviewed strategic planning processes to assess the extent to which they impact on realising alignment towards a collective health sector objective. Employing qualitative approaches, we reviewed strategic plans under implementation in the health sector and using an interview guide consisting of open‐ended questions, interviewed key informants at the national and district level. Results showed that strategic plans are too many with majority of program strategies not well aligned to the health sector strategic plan, are not costed, and there overlaps in objectives among the several strategies addressing the same program. Weaknesses in the development process, perceived poor quality of the strategies, limited capacity, high staff turnover, and inadequate funding were the identified challenges that abate the utility of the strategic plans. Moving towards UHC starts with a robust planning process that rallies all actors and all available resources around a common objective. The planning process should be strengthened through ensuring participatory processes, evidence informed prioritisation, MoH institutional capacity to lead the process, and consideration for implementation feasibility. Flexibility to take into consideration emerging evidence and new developments in global health needs consideration.
Inhibiting homologous recombination decreases extrachromosomal amplification but has no effect on intrachromosomal amplification in methotrexate‐resistant colon cancer cells
Authors: Cai Mengdi , Zhang Huishu , Hou Liqing , Gao Wei , Song Ying , Cui Xiaobo , Li Chunxiang , Guan Rongwei , Ma Jinfa , Wang Xu , Han Yue , Lv Yafan , Chen Feng , Wang Ping , Meng Xiangning , Fu Songbin
Published: August 2018

Gene amplification, which involves the two major topographical structures double minutes (DMs) and homegeneously stained region (HSR), is a common mechanism of treatment resistance in cancer and is initiated by DNA double‐strand breaks. NHEJ, one of DSB repair pathways, is involved in gene amplification as we demonstrated previously. However, the involvement of homologous recombination, another DSB repair pathway, in gene amplification remains to be explored. To better understand the association between HR and gene amplification, we detected HR activity in DM‐ and HSR‐containing MTX‐resistant HT‐29 colon cancer cells. In DM‐containing MTX‐resistant cells, we found increased homologous recombination activity compared with that in MTX‐sensitive cells. Therefore, we suppressed HR activity by silencing BRCA1, the key player in the HR pathway. The attenuation of HR activity decreased the numbers of DMs and DM‐form amplified gene copies and increased the exclusion of micronuclei and nuclear buds that contained DM‐form amplification; these changes were accompanied by cell cycle acceleration and increased MTX sensitivity. In contrast, BRCA1 silencing did not influence the number of amplified genes and MTX sensitivity in HSR‐containing MTX‐resistant cells. In conclusion, our results suggest that the HR pathway plays different roles in extrachromosomal and intrachromosomal gene amplification and may be a new target to improve chemotherapeutic outcome by decreasing extrachromosomal amplification in cancer.
Performance evaluation of different precoding matrices for PAPR reduction in OFDM systems
Authors: Mounir Mohamed , Tarrad Ibrahim F. , Youssef Mohamed I.
Published: August 2018

Orthogonal frequency division multiplexing (OFDM) is an attractive technique for wireless communication systems due to its ability to mitigate frequency selectivity. However, OFDM suffers from high peak‐to‐average power ratio (PAPR) problem, which reduces the power amplifier (PA) efficiency or worse it degrades bit error rate (BER) performance and increases out‐of‐band radiation. In literature, there are different PAPR reduction techniques. Among them PAPR reduction by precoding matrices has small computational complexity along with high PAPR reduction gain. In this paper the six precoding matrices found in the literature are compared. Results showed that, all precoding matrices, except the one that based on square‐root raised cosine function (SRC), are not effective in terms of BER performance in presence of nonlinear PA, especially in high modulation order schemes (eg, 16‐QAM and 64‐QAM). However, precoding technique based on SRC matrix requires high data rate loss, to enhance the BER performance in presence of nonlinear PA. So, it can be said that, precoding technique worsens the problem especially in high modulation order schemes, except the SRC matrix‐based precoding technique, which cannot be used without high data rate loss.
In vitro assays and techniques utilized in anticancer drug discovery
Authors: Ediriweera Meran Keshawa , Tennekoon Kamani Hemamala , Samarakoon Sameera Ranganath
Published: August 2018

Development of a cancer is a multistep process and six major hallmarks of cancer that are known to control malignant transformation have been described. Anticancer drug development is a tedious process, requiring a number of in vitro, in vivo and clinical studies. In vitro assays provide an initial platform for cancer drug discovery approaches. A wide range of in vitro assays/techniques have been developed to evaluate each hallmark feature of cancer and selection of a particular in vitro assay or technique mainly depends on the specific research question (s) to be examined. In the present review, we have described some commonly utilized in vitro assays and techniques used to examine cell viability/proliferation, apoptosis, cellular senescence, invasion and migration, oxidative stress and antioxidant effects, gene and protein expression, angiogenesis and genomic alterations in cancer drug discovery. Additionally, uses of modern techniques such as high throughput screening, high content screening and reporter gene assays in cancer drug discovery have also been described.
Increasing the tissue thickness at implant sites using guided bone regeneration and an additional collagen matrix: Histologic observations in beagle dogs
Authors: Seo Gi‐Young , Thoma Daniel Stefan , Jung Ui‐Won , Lee Jung‐Seok
Published: August 2018

Objectives

To histologically determine the alteration in horizontal mucosal thickness at sites that received guided bone regeneration (GBR) with additional use of collagen matrix and to assess whether bone formation is affected by adding collagen matrix at GBR sites at 8 weeks of healing.

Materials and methods

Eight weeks after bilateral extraction of maxillary premolars, standardized defects were created on buccal side of edentulous ridges in four beagle dogs. One side was randomly allocated as control (biphasic calcium phosphate plus collagen membrane; GBR only), while contralateral side was allocated as test (biphasic calcium phosphate plus collagen membrane plus an additional layer of collagen matrix). Histologic observations, histomorphometric and micro‐computed tomography analyses were performed after 8 weeks.

Results

Membrane complex comprising residual collagen membrane and adjacent dense connective tissue was observed at both control and test sites. The thickness in the histologic analysis were 1.69 ± 0.23 mm (control) and 1.76 ± 0.07 mm (test) in histologic analysis and were 2.03 ± 0.26 mm (control) and 2.14 ± 0.24 mm (test) in radiographic analysis. The thickness of the membrane complex in soft‐tissue layer were 723.0 ± 241.6 μm (control) and 984.6 ± 334.4 μm (test). The percentage of new bone formation were 22.30 ± 5.92% (control) and 25.50 ± 8.08% (test). All measured outcome did not show significant differences between control and test groups.

Conclusion

The addition of collagen matrix on top of standard GBR procedure did not increase the soft tissue thickness and dense connective tissue formation at 8 weeks of healing. Bone regeneration was not affected by the addition of collagen matrix. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 741–749, 2019.
Paeoniflorin inhibits mast cell–mediated allergic inflammation in allergic rhinitis
Authors: Wang Guanghui , Cheng Nan
Published: August 2018

Paeoniflorin (PF), one of the main effective ingredients from the root of Paeonia lactiflora Pall., was reported to possess antitumor, anti‐inflammatory, and antiallergic properties. However, the roles of PF in activated human mast cell line, HMC‐1 cells, have not yet been elucidated. Thus, the aim of this study was to examine the antiallergic and anti‐inflammatory effects of PF on phorbol‐12‐myristate 13‐acetate plus calcium ionophore (PMACI)–induced human mast cells and to identify the mechanism responsible for these effects. Our results demonstrated that pretreatment with PF effectively attenuated PMACI‐induced production of tumor necrosis factor‐α and interleukin 1β in HMC‐1 cells. In addition, PF significantly suppressed PMACI‐induced histamine release and caspase‐1 activation in HMC‐1 cells. Furthermore, PF prevented the activation of nuclear factor κB (NF‐κB) and mitogen‐activated protein kinase (MAPK) signaling pathways in activated HMC‐1 cells. In conclusion, we showed for the first time that PF attenuated the mast cell–mediated allergic inflammatory response through suppressing the NF‐κB and MAPK signaling pathways.
The establishment of clonally derived chicken embryonic fibroblast cell line (CSC) with high transfection efficiency and ability as a feeder cell
Authors: Zhao Ruifeng , Jin Jing , Sun Xinyu , Jin Kai , Wang Man , Ahmed Mahmoud F. , Zuo Qisheng , Zhang Yani , Zhao Zhenhua , Chen Guohong , Li Bichun
Published: August 2018

This study established a single cloned chicken embryonic fibroblast (CEF) cell line. It solves the main problem of the instability of a cultured primary cell and its impact on the experiment. In this study, CEF pass through this crisis and formed a continuous cell line after subculture. We isolated single postcrisis CEF by a mouth pipette under a convert microscope then established a single cloned cell line named CSC‐1‐5 which passaged continuously from 96‐well plates to 60 mm culture plates. CSC has a normal chicken diploid karyotype, no tumorigenicity, and a high G2/M phase cell ratio. We found that Fugene could mediate the transfection of CSCs efficiently; it was significantly improved compared with the primary cells. It could also promote the proliferation of chicken embryonic stem cell as a feeder layer.
Identification of novel heterozygous Apex 1 gene variant (Glu87Gln) in patients with head and neck cancer of Indian origin
Authors: Abarna R , Dutta Debnarayan , Sneha P , George Priya Doss C , Anbalagan M
Published: August 2018

Gene polymorphism among humans is one of the factors governing individual’s susceptibility and resistance to various diseases including cancer. DNA repair enzymes play an important role in protecting our genome from various mutagens and preventing cancer. The role of DNA repair enzyme Apurinic/Apyrimidinic endodeoxyribonuclease 1 (Apex 1) in cancer has been very well documented. Using genomic DNA, Apex 1 coding region of 76 patients (n = 76) with head and neck cancer were amplified and sequenced to detect variations in the sequence. Of 76 patients, 1 patient with heterozygous novel Apex 1 variant (Glu87Gln) was identified. A comparative analysis of wild type and variant protein using in silico approach was performed to understand the difference in the structure and the function. This further revealed that the variant had a slight impact on the structure, which affected the stability and function of the protein. Using the state‐of‐the‐art Molecular dynamic simulation analysis, we observed a loss in number of hydrogen bonds and salt bridge with a substitution of Gln for Glu at Position 87. This could be a possible reason behind the loss of stability/function of the protein. This study revealed a new variant of the Apex 1 gene; further studies will lead to the novel roles played by the variant Apex 1 protein in cause, disease progression, and response to the treatment in patients with cancer with Glu87Gln variant.
Overexpression of dedicator of cytokinesis 2 correlates with good prognosis in colorectal cancer associated with more prominent CD8+ lymphocytes infiltration: a colorectal cancer analysis
Authors: Miao Sen , Zhang Ren‐Ya , Wang Wei , Wang Hong‐Bo , Meng Li‐Li , Zu Li‐Dong , Fu Guo‐Hui
Published: August 2018

Recently, dedicator of cytokinesis 2 (DOCK2) has been reportedly exhibited high mutation prevalence in the Asian colorectal cancer (CRC) cohort. However, the expression pattern of DOCK2 and its clinical significance in CRC were still unknown. To characterize the role of DOCK2, a tissue microarray (TMA) containing 481 archived paraffin‐embedded CRC specimens was performed by immunohistochemistry. Among which, 54 primary CRC tissues showed high expression of DOCK2 protein, while others were negative. Moreover, DOCK2 expression was positively associated with invasion depth (P < .001) and tumor size (P = .016). Significantly, Kaplan‐Meier survival analysis revealed that patients with higher DOCK2 expression had a longer overall survival time (P = .017). Furthermore, univariate and multivariate Cox regression analysis confirmed that DOCK2 is an independent prognostic marker in CRC (P = .049,; HR, 0.519; 95% CI, 0.270 to 0.997). In addition, we observed a strong correlation between the infiltration of CD8+ T lymphocytes and DOCK2 expression (P = .0119). Our findings demonstrated that overexpressed DOCK2 might involve in recruiting CD8+ T lymphocytes and serve as a novel prognostic indicator and indicated a potential therapeutic strategy by restoring DOCK2 for CRC.
Oncostatin M–induced blood‐brain barrier impairment is due to prolonged activation of STAT3 signaling in vitro
Authors: Takata Fuyuko , Dohgu Shinya , Matsumoto Junichi , Machida Takashi , Sakaguchi Shinya , Kimura Ikuya , Yamauchi Atsushi , Kataoka Yasufumi
Published: August 2018

Oncostatin M (OSM) is a member of the interleukin (IL)‐6 family cytokines. We previously demonstrated that OSM induces blood‐brain barrier (BBB) impairment. However, functional characterization of IL‐6 family cytokines in BBB regulation and the cytokine‐related intracellular signaling pathway remain unclear. In this study, we demonstrate that among IL‐6 family cytokines, including IL‐6 and leukemia inhibitory factor (LIF), OSM is the most potent molecule for inducing BBB dysfunction via prolonged activation of signal transducer and activator of transcription (STAT) 3 following Janus‐activated kinase (JAK) activation. OSM but not IL‐6 and LIF (100 ng/mL for 24 hours) markedly produced increased sodium fluorescein permeability and decreased transendothelial electrical resistance in rat brain endothelial cell (RBEC) monolayers. This OSM‐induced BBB dysfunction was accompanied by decreased levels of claudin‐5 expression in RBECs, which were ameliorated by JAK inhibitor. We examined the time‐course of STAT3 phosphorylation in RBECs treated with OSM, IL‐6, and LIF. OSM upregulated STAT3 phosphorylation levels during a 24 hours period with a peak at 10 minutes. While IL‐6 and LIF transiently increased phosphorylated STAT3 at 10 minutes after addition, this phosphorylation decreased during the period from 1 to 24 hours after addition. These findings suggest that OSM‐induced sustained increases in STAT3 phosphorylation levels largely contribute to BBB impairment. Thus, elevated OSM levels and activation of brain endothelial JAK/STAT3 signaling pathway under pathological conditions should be considered as a possible hallmark for induction and development of BBB impairment.
Iron metabolism gene expression and prognostic features of hepatocellular carcinoma
Authors: Shen Ying , Li Xin , Zhao Bin , Xue Yanru , Wang Shenghang , Chen Xin , Yang Jiancheng , Lv Huanhuan , Shang Peng
Published: August 2018

Iron metabolism is crucial to hepatocellular carcinoma progression and is a key determinant of prognosis. Protein‐protein interactions within the iron metabolism gene network were analyzed using the European Molecular Biology Laboratory's Search Tool for Recurring Instances of Neighbouring Genes/Proteins database. We obtained 423 liver hepatocellular carcinoma gene expression profiles from the Cancer Genome Atlas database. The expression and pathway enrichment of representative iron intake genes (TFRC and DMT1), utilization genes (FTH1, FTL, HIF1A, HMOX1, SLC25A37, and SLC25A38), and efflux genes (FLVCR1 and SLC40A1) was investigated in tumor and adjacent tissues. We determined the relationship between iron metabolism and the prognostic features of liver hepatocellular carcinoma. The liver metabolism genes TFRC and FLVCR1 were related to survival, disease status, and prognosis in patients with hepatocellular carcinoma. Our results provide novel insight into liver cancer therapy.
A prognostic 3‐long noncoding RNA signature for patients with gastric cancer
Authors: Cheng Peng
Published: August 2018

Current studies showed that long noncoding RNAs (lncRNAs) may act as prognostic biomarkers in a variety of cancers. The aim of this study was to identify and assess a prognostic lncRNA signature in patients with gastric carcinoma (GC). LncRNAs expression profiles and corresponding clinicopathological data for 350 patients with GC were obtained from The Cancer Genome Atlas (TCGA), and the least absolute shrinkage and selection operator Cox (LASSO Cox) regression model was used to identify the lncRNA signature. Finally, 3 lncRNAs (CYP4A22‐AS1, AP000695.6, and RP11‐108M12.3) were identified using LASSO Cox. The prognostic score was imputed as follows: (0.354 × expression level of AP000695.6) + (− 0.899 × expression level of CYP4A22‐AS1) + (0.881 × expression level of RP11‐108M12.3). The nomogram that integrated independent prognostic factors (age, American Joint Committee on Cancer‐lymph node status, and residual tumor and risk score) was constructed. In TCGA cohort, the area under the curve (AUC) for the predictive nomogram was 0.737 (19‐month survival). The calibration curve also demonstrated satisfactory agreement between predictive values and observation values in the probabilities of 1‐, 3‐, and 5‐year overall survival. We also established prognostic model using machine learning technique, and the corresponding AUC for the predictive model was 0.756 (19‐month survival). The Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the Hippo signaling pathway was the main pathway associated with the 3‐lncRNA signature.
Inhibition of transforming growth factor‐β signaling pathway enhances the osteogenic differentiation of unrestricted somatic stem cells
Authors: Hedayati Sara , Parvaneh Tafreshi Azita , Moradi Nahid , Zeynali Bahman
Published: August 2018

In recent years, extensive studies have been performed to enhance stem cell‐based therapies for bone and cartilage repair. Among various sources of stem cells, cord blood‐derived unrestricted somatic stem cells (USSCs) seem to be the most appropriate option for an autologous transplantation. Among different signaling pathways, the transforming growth factor‐β (TGF‐β) pathway is shown as an important regulator of proliferation and osteogenic differentiation in osteoblast progenitors as well as mesenchymal stem cells. Due to its contradictory and temporally variable effects on different cell types, we sought to investigate whether and how the TGF‐β signaling pathway regulates the osteogenic differentiation of the USSCs. Therefore, in the current study, we treated USSCs with the recombinant protein TGF‐β1 (1 ng/mL) and showed that the expression of matrix metalloproteinase 9, a well‐known effector in this pathway, was significantly induced, indicating that the TGF‐β signaling pathway is active in USSCs. Then we applied a TGF‐β receptor antagonist (SB431542; 10 μM) to the osteogenic media cultured USSCs for single periods of 3.5 days within the 21‐day differentiation period starting at day 0, 3.5, 7, 10.5, 14, and 17.5. The expression analysis results of the of the osteogenic marker runt‐related transcription factor 2 as well as the production of bone matrix showed that SB431542 induced the osteogenic differentiation of USSCs more significantly during the early stage of differentiation, suggesting that the TGF‐β pathway temporally regulates the osteogenic differentiation of USSCs.
Sodium–glucose cotransporter 2 inhibitors and inflammation in chronic kidney disease: Possible molecular pathways
Authors: Yaribeygi Habib , Butler Alexandra E. , Atkin Stephen L. , Katsiki Niki , Sahebkar Amirhossein
Published: August 2018

Clinical trials with sodium–glucose cotransporter 2 (SGLT2) inhibitors (empagliflozin, dapagliflozin, and canagliflozin) have shown a decrease in the progression of chronic kidney disease (CKD). SGLT2 inhibitors represent a new category of oral antidiabetic agents that can also reduce systolic and diastolic blood pressure, as well as serum uric acid, and improve the glomerular filtration rate. Apart from affecting renal hemodynamics and glycotoxicity, evidence suggests that SGLT2 inhibitors may be renoprotective due to their effects on inflammation in renal tissues. Inflammatory responses play a prominent role in the pathophysiology of CKD as several structural and functional disorders of renal failure are strongly related to the overproduction of proinflammatory mediators. The present review discusses the anti‐inflammatory properties of SGLT2 inhibitors. The different molecular pathways through which SGLT2 inhibitors may affect inflammation in the kidneys are also commented upon.
Selection of reference genes for quantitative real‐time polymerase chain reaction studies in rat osteoblasts
Authors: Abuna Rodrigo P. F. , Oliveira Fabiola S. , Ramos Jaqueline I. R. , Lopes Helena B. , Freitas Gileade P. , Souza Alann T. P. , Beloti Marcio M. , Rosa Adalberto L.
Published: August 2018

Quantitative real‐time polymerase chain reaction (qRT‐PCR) is a powerful tool to evaluate gene expression, but its accuracy depends on the choice and stability of the reference genes used for normalization. In this study, we aimed to identify reference genes for studies on osteoblasts derived from rat bone marrow mesenchymal stem cells (bone marrow osteoblasts), osteoblasts derived from newborn rat calvarial (calvarial osteoblasts), and rat osteosarcoma cell line UMR‐106. The osteoblast phenotype was characterized by ALP activity and extracellular matrix mineralization. Thirty‐one candidates for reference genes from a Taqman® array were assessed by qRT‐PCR, and their expressions were analyzed by five different approaches. The data showed that several of the most traditional reference genes, such as Actb and Gapdh, were inadequate for normalization and that the experimental conditions may affect gene stability. Eif2b1 was frequently identified among the best reference genes in bone marrow osteoblasts, calvarial osteoblasts, and UMR‐106 osteoblasts. Selected stable and unstable reference genes were used to normalize the gene expression of Runx2, Alp, and Oc. The data showed statistically significant differences in the expression of these genes depending on the stability of the reference gene used for normalization, creating a bias that may induce incorrect assumptions in terms of osteoblast characterization of these cells. In conclusion, our study indicates that a rigorous selection of reference genes is a key step in qRT‐PCR studies in osteoblasts to generate precise and reliable data.
Novel role of Giα2 in cell migration: Downstream of PI3‐kinase–AKT and Rac1 in prostate cancer cells
Authors: Caggia Silvia , Chunduri HimaBindu , Millena Ana C. , Perkins Jonathan N. , Venugopal Smrruthi V. , Vo BaoHan T. , Li Chunliang , Tu Yaping , Khan Shafiq A.
Published: August 2018

Tumor cell motility is the essential step in cancer metastasis. Previously, we showed that oxytocin and epidermal growth factor (EGF) effects on cell migration in prostate cancer cells require Giα2 protein. In the current study, we investigated the interactions among G‐protein coupled receptor (GPCR), Giα2, PI3‐kinase, and Rac1 activation in the induction of migratory and invasive behavior by diverse stimuli. Knockdown and knockout of endogenous Giα2 in PC3 cells resulted in attenuation of transforming growth factor β1 (TGFβ1), oxytocin, SDF‐1α, and EGF effects on cell migration and invasion. In addition, knockdown of Giα2 in E006AA cells attenuated cell migration and overexpression of Giα2 in LNCaP cells caused significant increase in basal and EGF‐stimulated cell migration. Pretreatment of PC3 cells with Pertussis toxin resulted in attenuation of TGFβ1‐ and oxytocin‐induced migratory behavior and PI3‐kinase activation without affecting EGF‐induced PI3‐kinase activation and cell migration. Basal‐ and EGF‐induced activation of Rac1 in PC3 and DU145 cells were not affected in cells after Giα2 knockdown. On the other hand, Giα2 knockdown abolished the migratory capability of PC3 cells overexpressing constitutively active Rac1. The knockdown or knockout of Giα2 resulted in impaired formation of lamellipodia at the leading edge of the migrating cells. We conclude that Giα2 protein acts at two different levels which are both dependent and independent of GPCR signaling to induce cell migration and invasion in prostate cancer cells and its action is downstream of PI3‐kinase–AKT–Rac1 axis.
Effects of biphasic and monophasic electrical stimulation on mitochondrial dynamics, cell apoptosis, and cell proliferation
Authors: Love Maria R. , Sripetchwandee Jirapas , Palee Siripong , Chattipakorn Siriporn C. , Mower Morton M. , Chattipakorn Nipon
Published: August 2018

Currently, electrical stimulation (ES) is used to induce changes in various tissues and cellular processes, but its effects on mitochondrial dynamics and mechanisms are unknown. The aim of this study was to compare the effects of monophasic and biphasic, anodal, and cathodal ES on apoptosis, proliferation, and mitochondrial dynamics in neuroblastoma SH‐SY5Y cells. Cells were cultured and treated with ES. Alamar blue assay was performed to measure cell proliferation. The proteins expression of apoptotic‐related proteins Bcl‐2 associated X (Bax), B cell lymphoma 2 (Bcl‐2), optic‐atrophy‐1 (OPA1), mitofusin2 (Mfn2), phosphorylated dynamin‐related protein 1 at serine 616 (p‐DRP1), and total dynamin‐related protein 1 (Total‐DRP1) were also determined. The results showed that monophasic anodal and biphasic anodal/cathodal (Bi Anod) ES for 1 hr at 125 pulses per minute (2.0 Hz) produced the most significant increase in cell proliferation. In addition, monophasic anodal and Bi Anod ES treated cells displayed a significant increase in the levels of anti‐apoptotic protein Bcl‐2, whereas the Bax levels were not changed. Moreover, the levels of Mfn2 were increased in the cells treated by Bi Anod, and OPA1 was increased by monophasic anodal and Bi Anod ES, indicating increased mitochondrial fusion in these ES‐treated cells. However, the levels of mitochondrial fission indicated by DRP1 remained unchanged compared with non‐stimulated cells. These findings were confirmed through visualization of mitochondria using Mitotracker Deep Red, demonstrating that monophasic anodal and Bi Anod ES could induce pro‐survival effects in SH‐SY5Y cells through increasing cell proliferation and mitochondrial fusion. Future research is needed to validate these findings for the clinical application of monophasic anodal and Bi Anod ES.
Biomedical applications of carbon nanomaterials: Drug and gene delivery potentials
Authors: Mohajeri Mohammad , Behnam Behzad , Sahebkar Amirhossein
Published: August 2018

One of the major components in the development of nanomedicines is the choice of the right biomaterial, which notably determines the subsequent biological responses. The popularity of carbon nanomaterials (CNMs) has been on the rise due to their numerous applications in the fields of drug delivery, bioimaging, tissue engineering, and biosensing. Owing to their considerably high surface area, multifunctional surface chemistry, and excellent optical activity, novel functionalized CNMs possess efficient drug‐loading capacity, biocompatibility, and lack of immunogenicity. Over the past few decades, several advances have been made on the functionalization of CNMs to minimize their health concerns and enhance their biosafety. Recent evidence has also implied that CNMs can be functionalized with bioactive peptides, proteins, nucleic acids, and drugs to achieve composites with remarkably low toxicity and high pharmaceutical efficiency. This review focuses on the three main classes of CNMs, including fullerenes, graphenes, and carbon nanotubes, and their recent biomedical applications.
Analysis of pro‐ and anti‐inflammatory cytokine gene variants and serum cytokine levels as prognostic markers in breast cancer
Authors: Kaur Raman Preet , Vasudeva Kanika , Singla Heena , Benipal Raja Paramjeet Singh , Khetarpal Preeti , Munshi Anjana
Published: August 2018

The aim of current study was to evaluate the genetic variation in all the genes encoding pro‐ and anti‐inflammatory cytokines in association with breast cancer development in patients from Malwa region of Punjab. The importance of the levels of interleukin (IL)‐17, tumor necrosis factor, interferon γ, IL‐10, IL‐6, IL‐4, and IL‐2 with respect to clinicopathological data, prognosis, and disease‐free survival was also determined in these patients. Two hundred and fifty female breast cancer patients and 250 age‐matched controls were screened for variations in cytokine‐encoding genes using global screening array microchip and PCR‐RFLP. The level of cytokines was estimated in 150 patients and 60 age‐matched controls using BD™ Cytometric Bead Array (CBA) Human Th1/Th2/Th17 cytokine kit by BD Accuri flow cytometer. The difference in cytokine levels was evaluated by Mann–Whitney test. No significant variation in the genes encoding various cytokines was found between patients and controls. Out of the seven cytokines evaluated, the levels of IL‐6 and IL‐17a were found to be significantly high in patients in comparison with controls (
p = 0.001 and 0.02, respectively). The elevated levels of these cytokines are also associated significantly with poor outcome. We did not find any specific variation in the genes encoding various cytokines between patients and controls. However, there was a significant difference in the serum levels of IL‐6 and IL‐17a between patients and controls, and the elevated levels of these two cytokines associated significantly with poor outcome in breast cancer patients and, therefore, can be used as prognostic markers.
Endurance training prevents inflammation and apoptosis in hypothalamic neurons of obese mice
Authors: Marinho Rodolfo , Munõz Vitor R. , Pauli Luciana S. S. , Ropelle Eloize C. C. , de Moura Leandro P. , Moraes Juliana C. , Moura‐Assis Alexandre , Cintra Dennys E. , da Silva Adelino S. R. , Ropelle Eduardo R. , Pauli José R.
Published: August 2018

This study investigated the effects of exercise training in regulating inflammatory processes, endoplasmic reticulum stress, and apoptosis in hypothalamic neurons of obese mice. Swiss mice were distributed into three groups: Lean mice (Lean), sedentary animals fed a standard diet; obese mice (Obese), sedentary animals fed a high‐fat diet (HFD); trained obese mice (T. Obese), animals fed with HFD and concurrently subjected to an endurance training protocol for 8 weeks. In the endurance training protocol, mice ran on a treadmill at 60% of peak workload for 1 hr, 5 days/week for 8 weeks. Twenty‐four hours after the last exercise session, the euthanasia was performed. Western blot, quantitative real‐time polymerase chain reaction, and terminal deoxynucleotide transferase biotin‐dUTP nick end‐labeling (TUNEL) techniques were used for the analysis of interest. The results show exercise training increased phosphorylation of leptin signaling pathway proteins (pJAK2/pSTAT3) and reduced the content of tumor necrosis factor α, toll‐like receptor 4, suppressor of cytokine signaling 3, protein–tyrosine phosphatase 1B as well as the phosphorylation of IkB kinase in the hypothalamus of T. Obese animals. A reduction of macrophage activation and phosphorylation of eukaryotic initiation factor 2α, and protein kinase RNA‐like endoplasmic reticulum kinase (PERK) were also observed in exercised animals. Furthermore, exercise decreased the expression of the proapoptotic protein (PARP1) and increased anti‐inflammatory (IL‐10) and antiapoptotic (Bcl2) proteins. Using the TUNEL technique, we observed that the exercised animals had lower DNA fragmentation. Finally, physical exercise preserved pro‐opiomelanocortin messenger RNA content. In conclusion, exercise training was able to reorganize the control of the energy balance through anti‐inflammatory and antiapoptotic responses in hypothalamic tissue of obese mice.
MALAT1 induces osteosarcoma progression by targeting miR‐206/CDK9 axis
Authors: Ren Dong , Zheng Hao , Fei Sang , Zhao Jia‐Li
Published: August 2018

Long noncoding RNAs (LncRNAs) have been reported to participate in cancer development, including osteosarcoma. Here, in our study, we observed that lncRNA metastasis‐associated lung adenocarcinoma transcription 1 (MALAT1) was remarkably overexpressed in osteosarcoma. However, the role it plays in osteosarcoma proliferation mediated by miR‐206/cyclin‐dependent kinase 9 (CDK9) axis remains uninvestigated. It was found that miR‐206 was decreased and CDK9 was elevated in human osteosarcoma cells including MG63, Saos‐2, U2OS, and KHOS compared with human osteoblast cell line hFOB 1.19. In addition, it was exhibited that knockdown of MALAT1 was able to inhibit osteosarcoma cell proliferation, which suggested that MALAT1 played an oncogenic role in osteosarcoma development. Bioinformatics analysis indicated that MALAT1 can function as a competing endogenous RNA by sponging miR‐206. Because miR‐206 has been identified as a significant tumor suppressive gene in multiple cancers, we validated that mimics of miR‐206 can restrain osteosarcoma progression. Furthermore, dual‐luciferase reporter assay, RNA binding protein immunoprecipitation, and RNA pull‐down assay demonstrated the correlation between miR‐206 and MALAT1. Besides these, CDK9 was predicted as a downstream gene of miR‐206, and we observed that MALAT1 can regulate osteosarcoma progress by modulating CDK9 expression via sponging miR‐206. In conclusion, our study implied that MALAT1/miR‐206/CDK9 axis can provide novel insights into the biological mechanism of osteosarcoma progression.
Investigation on silent bacterial infections in specimens from pregnant women affected by spontaneous miscarriage
Authors: Contini Carlo , Rotondo John C. , Magagnoli Federica , Maritati Martina , Seraceni Silva , Graziano Angela , Poggi Alice , Capucci Roberta , Vesce Fortunato , Tognon Mauro , Martini Fernanda
Published: August 2018

Miscarriage is one of the main complications occurring in pregnancy. The association between adverse pregnancy outcomes and silent bacterial infections has been poorly investigated.Ureaplasma parvumandurealiticum, Mycoplasma genitaliumandhominisandChlamydia trachomatisDNA sequences have been investigated by polymerase chain reaction (PCR) methods in chorionic villi tissues and peripheral blood mononuclear cells (PBMCs) from females with spontaneous abortion (SA,n = 100) and females who underwent voluntary interruption of pregnancy (VI,n = 100).U. parvumDNA was detected in 14% and 15% of SA and VI, respectively, with a mean of bacterial DNA load of 1.3 × 10−1copy/cell in SA and 2.8 × 10−3copy/cell in VI;U. urealiticumDNA was detected in 3% and 2% of SA and VI specimens, respectively, with a mean DNA load of 3.3 × 10−3copy/cell in SA and 1.6 × 10−3copy/cell in VI;M. hominisDNA was detected in 5% of SA specimens with a DNA load of 1.3 × 10−4copy/cell and in 6% of VI specimens with a DNA load of 1.4 × 10−4copy/cell;C. trachomatisDNA was detected in 3% of SA specimens with a DNA load of 1.5 × 10−4copy/cell and in 4% of VI specimens with a mean DNA load of 1.4 × 10−4copy/cell. In PBMCs from the SA and VI groups,Ureaplasma spp, Mycoplasma sppandC. trachomatisDNAs were detected with a prevalence of 1%–3%. Bacteria were investigated, for the first time, by quantitative real‐time PCR (qPCR) in chorionic villi tissues and PBMCs from women affected by SA and VI. These data may help to understand the role and our knowledge of the silent infections in SA.
Ferritinophagy/ferroptosis: Iron‐related newcomers in human diseases
Authors: Tang Mingzhu , Chen Zhe , Wu Di , Chen Linxi
Published: August 2018

Nuclear receptor coactivator 4 mediated ferritinophagy is an autophagic phenomenon that specifically involves ferritin to release intracellular free iron. Ferritinophagy is implicated in maintaining efficient erythropoiesis. Notably, ferritinophagy also plays a central role in driving some pathological processes, including Parkinson’s disease (PD) and urinary tract infections. Some evidence has demonstrated that ferritinophagy is critical to induce ferroptosis. Ferroptosis is a newly nonapoptotic form of cell death, characterized by the accumulation of iron‐based lipid reactive oxygen species. Ferroptosis plays an important role in inhibiting some types of cancers, such as hepatocellular carcinoma, pancreatic carcinoma, prostate cancer, and breast cancer. Conversely, the activation of ferroptosis accelerates neurodegeneration diseases, including PD and Alzheimer’s disease. Therefore, in this review, we summarize the regulatory mechanisms related to ferritinophagy and ferroptosis. Moreover, the distinctive effects of ferritinophagy in human erythropoiesis and some pathologies, coupled with the promotive or inhibitory role of tumorous and neurodegenerative diseases mediated by ferroptosis, are elucidated. Obviously, activating or inhibiting ferroptosis could be exploited to achieve desirable therapeutic effects on diverse cancers and neurodegeneration diseases. Interrupting ferritinophagy to control iron level might provide a potentially therapeutic avenue to suppress urinary tract infections.
Exercise ameliorates high‐fat diet‐induced impairment of differentiation of adipose‐derived stem cells into neuron‐like cells in rats
Authors: Kato Hisashi , Minamizato Hidemasa , Ohno Hideki , Ohira Yoshinobu , Izawa Tetsuya
Published: August 2018

Adipose‐derived stem cells (ADSCs) can differentiate into neurons under particular conditions. It remains largely unknown whether this differentiation potential is affected by physical conditions such as obesity, which modulates the functions of adipose tissue. In this study, we determined the impact of either a 9‐week high‐fat diet (60% fat; HFD) or 9‐week exercise training on the differentiation potential of ADSCs into neuron‐like cells in male Wistar rats. Rats were randomly assigned to a normal diet‐fed (ND‐SED) group, HFD‐fed (HFD‐SED) group, or exercise‐trained HFD‐fed group (HFD‐EX). After a 9‐week intervention, ADSCs from all groups differentiated into neuron‐like cells. Expression of neuronal marker proteins (nestin, βIII‐tubulin, and microtubule‐associated protein 2 [MAP2]) and the average length of cell neurites were lower in cells from HFD‐SED rats than in other groups. Instead, protein expression of COX IV and Cyt‐c, the Bax/Bcl‐2 and LC3‐II/I ratio, and the malondialdehyde level in culture medium were higher in cells from HFD‐SED rats. No significant difference between ND‐SED and HFD‐EX rats was observed, except for the average length of cell neurites in MAP2. Thus, HFD impaired the differentiation potential of ADSCs into neuron‐like cells, which was accompanied by increases in apoptotic activity and oxidative stress. Importantly, exercise training ameliorated the HFD‐induced impairment of neurogenesis in ADSCs. The adipose tissue microenvironment could influence the differentiation potential of ADSCs, a source of autologous stem cell therapy.
Nine hub genes as the potential indicator for the clinical outcome of diabetic nephropathy
Authors: Song Xiaoping , Gong Min , Chen Yanping , Liu Hui , Zhang Jun
Published: August 2018

Diabetic nephropathy (DN) is one of the most serious and dangerous chronic complications caused by diabetes mellitus, and the identification and development of novel biomarkers could be beneficial for the diagnosis and prognosis of DN patients. This study focused on researching the differently expressed pattern of the DN samples from glomeruli and tubulointerstitium. Significance analysis of microarrays (SAM) was used to identify differentially regulated genes in 44 microdissected human kidney samples. Functional enrichment analysis was used to analyze the functions these genes are mostly enriched in. Besides, protein–protein interaction (PPI) network was used to select the hub genes that were associated with DN. The gene expression pattern of DN samples from glomeruli and tubulointerstitium was found to be quite different. It showed that the recurrence rate of DN originating from glomeruli and control samples was lower than that from tubulointerstitium and control samples. A total of 332 differentially expressed genes were identified between glomeruli tissues and tubulointerstitium tissues. PPI network analysis was performed on these 332 genes. Finally, nine hub genes were selected as the most potential biomarkers in the occurrence of DN. In conclusion, a total of 332 genes were found to be related to DN, and these genes were found to be mostly enriched in pathways such as immunity, inflammatory, and vascular pathways. Three genes VAV1, LCK, and Plk had the potential to serve as indicators for the occurrence and development of DN in clinical management.
Monocyte‐to‐HDL‐cholesterol ratio as a prognostic marker in cardiovascular diseases
Authors: Ganjali Shiva , Gotto Antonio M. , Ruscica Massimiliano , Atkin Stephen L. , Butler Alexandra E. , Banach Maciej , Sahebkar Amirhossein
Published: August 2018

Inflammation and lipid accumulation are two basic hallmarks of atherosclerosis as a chronic disease. Inflammation not only is a local response but can also be considered as a systemic process followed by an elevation of inflammatory mediators. Monocytes are a major source of proinflammatory species during atherogenesis. In atherosclerosis, modified low‐density lipoproteins (LDLs) are removed by macrophages; these are recruited in the vessel wall, inducing the release of inflammatory cytokines in inflamed tissue. Hence, inflammatory cholesterol ester‐loaded plaque is generated. High‐density lipoprotein‐cholesterol (HDL‐C) exhibits antiatherosclerotic effects by neutralizing the proinflammatory and pro‐oxidant effects of monocytes via inhibiting the migration of macrophages and LDL oxidation in addition to the efflux of cholesterol from these cells. Furthermore, HDL plays a role in suppressing the activation of monocytes and proliferation–differentiation of monocyte progenitor cells. Thus, accumulation of monocytes and reduction of HDL‐C may participate in atherosclerosis and cardiovascular diseases (CVD). Given that the relationship between the high number of monocytes and low HDL‐C levels has been reported in inflammatory disorders, this review focused on understanding whether the monocyte‐to‐HDL ratio could be a convenient marker to predict atherosclerosis development and progression, hallmarks of CV events, instead of the individual monocyte count or HDL‐C level.
Infection‐associated epigenetic alterations in gastric cancer: New insight in cancer therapy
Authors: Fattahi Sadegh , Kosari‐Monfared Mohadeseh , Ghadami Elham , Golpour Monireh , Khodadadi Parastoo , Ghasemiyan Mohammad , Akhavan‐Niaki Haleh
Published: August 2018

Gastric cancer risk is higher for malignancies motivated by bacterial and viral infections. Epigenetic abnormalities including DNA methylation, histone modifications, and noncoding RNAs are important regulatory key players in gastric cancer development in infected patients. Epigenetic memory restoration is an extremely interesting phenomenon which should be considered in therapeutic approaches. In vitro and in vivo antiviral treatments in combination with epigenetic therapeutic strategies along with standard chemotherapy revealed promising outcomes in gastric cancer prevention and treatment. This review summarizes our current understanding of the gastric cancer infections and epigenetic alterations caused by these agents. We focus on studies highlighting recent advances in epigenetic restoration by target specific drugs and present also a comprehensive overview of effective antiviral drug treatments against gastric cancer.
Estrogen‐related receptor γ negatively regulates osteoclastogenesis and protects against inflammatory bone loss
Authors: Kim Hyun‐Ju , Kim Bo Kyung , Ohk Boram , Yoon Hye‐Jin , Kang Woo Youl , Cho Seungil , Seong Sook Jin , Lee Hae Won , Yoon Young‐Ran
Published: August 2018

Estrogen‐related receptor γ (ERRγ) is an orphan nuclear receptor that plays an important role in various metabolic processes under physiological and pathophysiological conditions. Here, we report that ERRγ functions as a negative regulator in receptor activator of nuclear factor κΒ ligand (RANKL)–induced osteoclast differentiation. We observed that ERRγ was strongly expressed in osteoclast precursors, bone marrow–derived macrophages (BMMs) while its expression was significantly reduced by RANKL during osteoclastogenesis. Overexpression of ERRγ in BMMs suppressed the formation of multinucleated osteoclasts and attenuated the induction of c‐Fos and nuclear factor of activated T cells c1, which are critical modulators in osteoclastogenesis. Similarly, the treatment of ERRγ agonists, N‐(4‐(diethylaminobenzylidenyl)‐N'‐(4‐hydroxybenzoyl)‐hydrazine (DY131) or GSK4716, also inhibited osteoclast generation and the expression of these key modulators. On the other hand, shRNA‐mediated knockdown of ERRγ accelerated the formation of bone‐resorbing cells and the expression of osteoclastogenic markers. Forced expression of ERRγ blocked RANKL‐stimulated phosphorylation of the nuclear factor κB (NF‐κB) inhibitor IκBα and suppressed NF‐κB transcriptional activity induced by RANKL or the NF‐κB subunit p65. Furthermore, by employing a pharmacological approach, we showed that the ERRγ agonist DY131 protected against inflammatory bone loss induced by lipopolysaccharide in vivo. Together, our findings reveal that ERRγ is a pivotal regulator in RANKL‐mediated osteoclastogenesis and suggest that ERRγ may have potential as a therapeutic target for pathological bone loss.