bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic Contributors and Modifiers of Biliary Atresia
Authors: Mezina Anya , Karpen Saul J.
Published: June 2015

To date, the etiology and pathogenic underpinning of the progression of the most prevalent serious neonatal liver disease, biliary atresia, remains elusive. This disease presents as an aggressive form of neonatal cholestasis characterized by the destruction and obliteration of the extrahepatic bile ducts within the first few weeks of life and a rapid progression of biliary fibrosis, likely due to unremitting cholestasis and retention of biliary constituents including bile acids. In ∼5% of patients, biliary atresia is associated with laterality features, suggesting a genetic underpinning to a disease that begins soon after birth. However, biliary atresia does not occur within families and twins are discordant, indicating an absence of strict mendelian inheritance. Despite this, genes related to bile duct dysmorphogenesis/ciliopathies overlapping with features of biliary atresia in both humans and nonhuman model systems have been proposed. Taken together, strict genetic etiologies leading to a common pathway of a neonatal cholangiopathy resulting in biliary atresia remain elusive. Contributions from fibrogenesis- and inflammation-based studies suggest that early engagement of these pathways contributes to disease progression, but a recent double-blind study did not suggest any benefit from early use of corticosteroids. However, there are genetic contributions to the adaptation and response to cholangiopathies and cholestasis that may be present in certain populations that likely impact upon the response to hepatoportoenterostomy and subsequent biliary tract function. Studies utilizing next generation sequencing technologies (e.g., exome analysis) are ongoing in several laboratories around the world; they are expected to provide insights into genetic contributions to biliary atresia outcomes. Altogether, combinations of exome sequencing and large population studies are expected to reveal causative and modifying genes relevant to patients with biliary atresia as a means to provide therapeutic targets and potential opportunities for genetic screening.
Nuclear Receptor Variants in Liver Disease
Authors: Zimmer Vincent , Liebe Roman , Lammert Frank
Published: June 2015

This snapshot reviews the current state of knowledge on genetic variants of nuclear receptors (NRs) involved in regulating various aspects of liver metabolism. Interindividual differences in responses to diet and other ‘in-' and environmental stressors can be caused by variants in components of the NR regulatory gene network. We recapitulate recent evidence for the application of NRs in genetic diagnosis of monogenic liver disease. Genetic analysis of multifactorial liver diseases, such as nonalcoholic fatty liver disease and diabetes mellitus, pinpoints key players in disease predisposition and progression. In particular, NR1H4 variants have been associated with intrahepatic cholestasis of pregnancy and gallstone disease. Other examples include studies of NR1I2 and NR1I3 polymorphisms in patients with drug-induced liver injury and NR5A2 variation in cholangiocarcinoma. Associations of NR gene variants have been identified in patients with dyslipidemia and other metabolic syndrome-associated traits by genome-wide studies. Evidence from these analyses confirms a role for NR variation in common diseases, linking regulatory networks to complex and variable phenotypes. These new insights into the impact of NR variants offer perspectives for their future use in diagnosis and treatment of common diseases.
TGR5 in the Cholangiociliopathies
Authors: Masyuk Tatyana V. , Masyuk Anatoliy I. , LaRusso Nicholas F.
Published: June 2015

A plasma membrane-bound G protein-coupled receptor, TGR5, that transmits bile acid signaling into a cellular response primarily via the cAMP pathway is expressed in human and rodent cholangiocytes and is localized to multiple, diverse subcellular compartments, including primary cilia. Ciliary-associated TGR5 plays an important role in cholangiocyte physiology and may contribute to a group of liver diseases referred to as the ‘cholangiociliopathies', which include polycystic liver disease (PLD) and, possibly, cholangiocarcinoma and primary sclerosing cholangitis. Based on our observations that (1) ciliated and nonciliated cholangiocytes respond to TGR5 activation differently (i.e. the level of cAMP increases in nonciliated cholangiocytes but decreases in ciliated cells) and (2) hepatic cysts are derived from cholangiocytes that are characterized by both malformed cilia and increased cAMP levels, we hypothesized that TGR5-mediated cAMP signaling in cystic cholangiocytes contributes to hepatic cystogenesis. Indeed, our studies show that TGR5 is overexpressed and mislocalized in cystic cholangiocytes, and when activated by ligands, results in increased intracellular cAMP levels, cholangiocyte hyperproliferation and cyst growth. Our studies also show that genetic elimination of TGR5 in an animal model of PLD inhibits hepatic cystogenesis. Collectively, these data suggest the involvement of TGR5 in PLD and that TGR5 targeting in cystic cholangiocytes may have therapeutic potential.
Bile Acid Signaling: Mechanism for Bariatric Surgery, Cure for NASH?
Authors: Kohli Rohit , Myronovych Andriy , Tan Brandon K. , Salazar-Gonzalez Rosa-Maria , Miles Lili , Zhang Wujuan , Oehrle Melissa , Sandoval Darleen A. , Ryan Karen K. , Seeley Randy J. , Setchell Kenneth D.R.
Published: June 2015

Bariatric surgery is the most effective and durable treatment option for obesity today. More importantly, beyond weight loss, bariatric procedures have many advantageous metabolic effects including reversal of obesity-related liver disease - nonalcoholic steatohepatitis (NASH). NASH is an important comorbidity of obesity given that it is a precursor to the development of liver cirrhosis that may necessitate liver transplantation in the long run. Simultaneously, we and others have observed increased serum bile acids in humans and animals that undergo bariatric surgery. Specifically, our preclinical studies have included experimental procedures such as ‘ileal transposition' or bile diversion and established procedures such as Roux-en-Y gastric bypass and the adjustable gastric band. Importantly, these effects are not simply the result of weight loss since our data show that the resolution of NASH and increase in serum bile acids are not seen in rodents that lose an equivalent amount of weight via food restriction. In particular, we have studied the role of altered bile acid signaling, in the potent impact of a bariatric procedure termed ‘vertical sleeve gastrectomy' (VSG). In this review we focus on the mechanisms of NASH resolution and weight loss after VSG surgery. We highlight the fact that bariatric surgeries can be used as ‘laboratories' to dissect the mechanisms by which these procedures work to improve obesity and fatty liver disease. We describe key bile acid signaling elements that may provide potential therapeutic targets for ‘bariatric-mimetic technologies' that could produce benefits similar to bariatric surgery - but without the surgery!
A Plaque-Type Solitary Reticulohistiocytoma in a Two-Year-Old Boy
Authors: Shibuya Rintaro , Tanizaki Hideaki , Kaku Yo , Yonezawa Masao , Ryu Yuri , Otsuka Atsushi , Kabashima Kenji , Miyachi Yoshiki
Published: June 2015

Reticulohistiocytoma (RH) is a dermal histiocytic infiltration composed of large histiocytes with eosinophilic glassy cytoplasm. RH is classified into three clinical forms: solitary RH, diffuse cutaneous RH without systemic involvement and multicentric reticulohistiocytosis with systemic diseases. Solitary RH generally manifests as a nodular lesion in adults without accompanying systemic diseases. Herein, we describe a case of solitary RH with an atypical clinical manifestation as a red-brown-colored plaque in a 2-year-old boy. Atypical presentations of RH may pose diagnostic difficulty unless RH is considered. A correct diagnosis of RH can ensure avoidance of unnecessary invasive procedures.
Rapid Bioassay for Detection of Thyroid-Stimulating Antibodies Using Cyclic Adenosine Monophosphate-Gated Calcium Channel and Aequorin
Authors: Araki Naohiro , Iida Mitsuru , Amino Nobuyuki , Morita Shinji , Ide Akane , Nishihara Eijun , Ito Mitsuru , Saito Jun , Nishikawa Tetsuo , Katsuragi Kiyonori , Miyauchi Akira
Published: June 2015

Background: Thyroid-stimulating antibodies (TSAb) are known to be responsible for hyperthyroidism in Graves' disease (GD). The conventional methods to measure TSAb depend on cell-based assays that require cumbersome procedures and a sterilized tissue culture technique. The aim of the present study was to develop a ready-to-use cell-based assay for measuring TSAb activity without requiring sterilized conditions. Methods: We developed a new assay kit using a frozen Chinese hamster ovary cell line expressing the thyroid-stimulating hormone receptor, cyclic adenosine monophosphate (cAMP)-gated calcium channel and aequorin, tentatively named the aequorin TSAb assay. Activated stimulatory G-protein-coupled adenylate cyclase increases intracellular cAMP, which then binds to the cyclic nucleotide-gated calcium channel. Activation of this channel allows Ca2+ to enter the cell, and the influx of Ca2+ can be measured with aequorin, which is quantified by a luminometer. Results can be obtained in only 4 h without sterilized conditions. TSAb activities were expressed by international units using the NIBSC 08/204 standard. Results: Positive results of aequorin TSAb were obtained in 197 of 199 (98.9%) of untreated patients with GD. Only 1 of 42 (2.3%) patients with painless thyroiditis had a weakly positive aequorin TSAb. All 45 patients with subacute thyroiditis and 185 normal subjects showed negative aequorin TSAb. As for chronic thyroiditis, all 52 euthyroid patients showed negative aequorin TSAb, but 8 of 50 (16.0%) hypothyroid patients had a positive reaction. However, these positive reactions were not induced by serum thyroid-stimulating hormone (TSH) and were thought to be induced by the stimulating activity of anti-TSH receptor immunoglobulins. Conventional porcine TSAb and Elecsys thyroid-stimulating hormone receptor antibodies were positive in 69.3 and 95.5% of GD, respectively. Conclusion: The aequorin TSAb assay was positive in 98.9% of GD and was more sensitive than the conventional assay. This assay can be conducted in only 4 h without sterilized conditions and is practically useful in general clinical laboratories.
Aspirin ‘Resistance': Impact on No-Reflow, Platelet and Inflammatory Biomarkers in Diabetics after ST-Segment Elevation Myocardial Infarction
Authors: Kuliczkowski Wiktor , Gasior Mariusz , Pres Damian , Kaczmarski Jacek , Laszowska Anna , Szewczyk Marta , Hawranek Michal , Tajstra Mateusz , Zeglen Slawomir , Polonski Lech , Serebruany Victor L.
Published: June 2015

Background: The no-reflow (NR) phenomenon exists despite percutaneous coronary intervention (PCI), and is especially prevalent in diabetics. The causes(s) of NR are not fully elucidated, but may be associated with impaired residual platelet and inflammatory reactivity during dual-antiplatelet therapy. Objective: To assess the relationship between dual-antiplatelet therapy, NR and conventional biomarkers suggestive of platelet and inflammatory response in diabetics following ST-segment elevation myocardial infarction (STEMI) treated with PCI. Methods: Sixty diabetics with (n = 27) and without NR (n = 33) were prospectively enrolled. All patients were treated with clopidogrel and aspirin. Platelet and inflammatory biomarkers were assessed serially in the peripheral blood and right atrium before and after PCI and then at 24 h, 7 days and 30 days. Results: Arachidonic acid (AA)-induced platelet aggregation and the serum thromboxane B2 level before and after PCI (in the peripheral and right atrium blood) were significantly higher in the NR patients than in those with no NR. AA-induced aggregation >100 (AUC*min) before PCI predicted NR in diabetic patients with 96.2% sensitivity and 38.5% specificity (AUC 0.66; 95% CI 0.52-0.71; p = 0.029). There were no other correlations between NR and platelet reactivity (collagen, adenosine diphosphate, thrombin receptor agonist peptide-induced aggregation, vasodilator-stimulated phosphoprotein platelet reactivity index, soluble P-selectin, soluble CD40 ligand, platelet-derived growth factor AB and the level of platelet-monocyte aggregates) or between NR and inflammatory indices (i.e. high-sensitivity C-reactive protein, interleukin 6 and interleukin 10). Conclusion: An inadequate response to aspirin, but not to clopidogrel, may be associated with the occurrence of the NR phenomenon in diabetics with STEMI who have been treated with primary PCI.
The Cholangiocyte Glycocalyx Stabilizes the ‘Biliary HCO3- Umbrella': An Integrated Line of Defense against Toxic Bile Acids
Authors: Maillette de Buy Wenniger Lucas J. , Hohenester Simon , Maroni Luca , van Vliet Sandra J. , Oude Elferink Ronald P. , Beuers Ulrich
Published: June 2015

Background: Destruction of cholangiocytes is the hallmark of chronic cholangiopathies such as primary biliary cirrhosis. Under physiologic conditions, cholangiocytes display a striking resistance to the high, millimolar concentrations of toxic bile salts present in bile. We recently showed that a ‘biliary HCO3- umbrella', i.e. apical cholangiocellular HCO3- secretion, prevents cholangiotoxicity of bile acids, and speculated on a role for extracellular membrane-bound glycans in the stabilization of this protective layer. This paper summarizes published and thus far unpublished evidence supporting the role of the glycocalyx in stabilizing the ‘biliary HCO3- umbrella' and thus preventing cholangiotoxicity of bile acids. Key Messages: The apical glycocalyx of a human cholangiocyte cell line and mouse liver sections were visualized by electron microscopy. FACS analysis was used to characterize the surface glycan profile of cultured human cholangiocytes. Using enzymatic digestion with neuraminidase the cholangiocyte glycocalyx was desialylated to test its protective function. Using lectin assays, we demonstrated that the main N-glycans in human and mouse cholangiocytes were sialylated biantennary structures, accompanied by high expression of the H-antigen (α1-2 fucose). Apical neuraminidase treatment induced desialylation without affecting cell viability, but lowered cholangiocellular resistance to bile acid-induced toxicity: both glycochenodeoxycholate and chenodeoxycholate (pKa ≥4), but not taurochenodeoxycholate (pKa <2), displayed cholangiotoxic effects after desialylation. A 24-hour reconstitution period allowed cholangiocytes to recover to a pretreatment bile salt susceptibility pattern. Conclusion: Experimental evidence indicates that an apical cholangiocyte glycocalyx with glycosylated mucins and other glycan-bearing membrane glycoproteins stabilizes the ‘biliary HCO3- umbrella', thus aiding in the protection of human cholangiocytes against bile acid toxicity.
A 14-Year Experience in the Management of Patients with Acquired Immune Thrombotic Thrombocytopenic Purpura in Northern Israel
Authors: Rinott Nadav , Mashiach Tatiana , Horowitz Netanel A. , Schliamser Liliana , Sarig Galit , Keren-Politansky Anat , Dann Eldad J.
Published: June 2015

Acquired idiopathic thrombotic thrombocytopenic purpura (I-TTP) is a life-threatening microangiopathic disorder usually treated with therapeutic plasma exchange (TPE). The current study assessed the role of rituximab in the treatment of complicated I-TTP. The sequence of TTP events was compared in a group of I-TTP patients treated with TPE and a cohort of refractory or relapsed patients who also received rituximab. This retrospective evaluation included 45 I-TTP patients, treated between January 2000 and October 2013, who underwent at least 3 TPE procedures and were followed up until December 2013 or death. Thirty-one patients with an uncomplicated course received TPE only. Fourteen patients had a complicated course due to either a primary refractory/exacerbated disease (n = 5) or relapse (n = 9) and received rituximab together with TPE. The median number of TPE procedures performed in the first TTP episode in the uncomplicated cohort and groups with primary refractory or relapsed TTP was 11, 27 and 45, respectively. The relapse rates per follow-up year in the uncomplicated I-TTP, primary refractory and relapsed I-TTP groups were 0.18, 0.2 and 0.6 episodes, respectively. After rituximab therapy this rate dropped to 0.2 per year in the relapsed subgroup. In conclusion, about a quarter of patients with I-TTP had a complicated course and experienced a major benefit from rituximab in terms of effectiveness and safety.
Preoperative Rosuvastatin Protects Patients with Coronary Artery Disease Undergoing Noncardiac Surgery
Authors: Xia Jinggang , Qu Yang , Yin Chunlin , Xu Dong
Published: June 2015

Objectives: We explored whether preoperative rosuvastatin could protect the cardiac health of patients with coronary artery disease undergoing emergency, noncardiac surgery. Methods: We randomized 550 noncardiac emergency surgery patients with stable coronary artery disease on long-term statin therapy to treatment with and without preoperative rosuvastatin. All patients received rosuvastatin after surgery. We evaluated the incidence of myocardial necrosis and major adverse cardiovascular and cerebrovascular events (MACCE) 30 days and 6 months after surgery. Results: Creatinine kinase-myocardial band (CK-MB) isoform elevations occurred less frequently 12 and 24 h after noncardiac emergency surgery in the experimental group than in the control group (p = 0.029). After surgery, the incidence of MACCE was also lower in the experimental group than in the control group (p = 0.019). The difference was mainly due to the incidence of perioperative myocardial infarction (p = 0.029). Multivariable analysis found that rosuvastatin reload reduced the incidence of MACCE 52% 6 months after surgery (p = 0.03). Conclusions: Preoperative rosuvastatin reload therapy decreases the incidence of myocardial necrosis and MACCE after noncardiac emergency surgery in patients with stable coronary artery disease on long-term statin therapy.
Ultrasonic Vocalizations by the Spectral Tarsier, Tarsius spectrum
Authors: Gursky Sharon
Published: June 2015

Although the vocalizations of spectral tarsiers have been studied for over 3 decades by numerous primatologists, the data in this paper represent the first evidence that this species communicates in the ultrasonic range. In addition, this paper characterizes the types of ultrasonic vocalizations by spectral tarsiers, Tarsius spectrum. Data were collected at Tangkoko Nature Reserve in Sulawesi, Indonesia, from January through April 2013. Recordings were made on a Wildlife Acoustics Ultrasonic Song Meter BAT2 from 10 groups of varying sizes and compositions. The ultrasonic recorder was placed at the base of the group's sleeping tree and recorded from 5.00 to 7.00 h using an omnidirectional microphone. The ultrasonic vocalizations fell into 5 main categories: chirps, twitters, choruses, doubles and whistles. Chirps were the most frequent ultrasonic vocalizations, followed by twitters, choruses, doubles and then whistles. While chirps, twitters and choruses extended from the audible to the ultrasonic range, the doubles and whistles were pure ultrasound. Currently, the function of these ultrasonic vocalizations is not yet clear and requires additional research.
Potential of nor-Ursodeoxycholic Acid in Cholestatic and Metabolic Disorders
Authors: Trauner Michael , Halilbasic Emina , Claudel Thierry , Steinacher Daniel , Fuchs Claudia , Moustafa Tarek , Pollheimer Marion , Krones Elisabeth , Kienbacher Christian , Traussnigg Stefan , Kazemi-Shirazi Lili , Munda Petra , Hofer Harald , Fickert Peter , Paumgartner Gustav
Published: June 2015

24-nor-ursodeoxycholic acid (norUDCA) is a side-chain shortened derivate of ursodeoxycholic acid (UDCA). Since norUDCA is only ineffectively conjugated with glycine or taurine, it has specific physicochemical and therapeutic properties distinct from UDCA. Nonamidated norUDCA undergoes cholehepatic shunting enabling ‘ductular targeting' and inducing a bicarbonate-rich hypercholeresis, with cholangioprotective effects. At the same time it has direct anti-inflammatory, antilipotoxic, anti fibrotic, and antiproliferative properties targeting various liver cell populations. norUDCA appears to be one of the most promising novel treatment approaches targeting the liver and the bile duct system at multifactorial and multicellular levels. This review article is a summary of a lecture given at the XXIII International Bile Acid Meeting (Falk Symposium 194) on ‘Bile Acids as Signal Integrators and Metabolic Modulators' held in Freiburg, October 8-9, 2014, and summarizes the recent progress with norUDCA as a novel therapeutic approach in cholestatic and metabolic (liver) disorders.
Laparoscopic Resection of Schwannoma of the Ascending Colon
Authors: Tashiro Yoshihiko , Matsumoto Fumio , Iwama Keiko , Shimazu Ai , Matsumori Sei , Nohara Shigeo , Miura Hiroyoshi , Takei Masahiko , Namekata Koji , Takase Masaru , Okada Motoi , Tsumura Hidenori
Published: June 2015

Schwannomas of the colon are rare and difficult to diagnose preoperatively. We report a case of schwannoma of the ascending colon that was resected laparoscopically. A 64-year-old woman was referred to our hospital by her local clinic for further evaluation and management of a submucosal tumor of the ascending colon. A definitive preoperative diagnosis could not be reached despite examinations. Gastrointestinal stromal tumor, leiomyoma and lymphoma were the differential diagnoses. We performed a laparoscopic right hemicolectomy with D2 lymph node dissection. Histological findings with hematoxylin-eosin staining revealed spindle-like tumor cells, and immunohistochemical analysis showed that the tumor was positive for S-100 but negative for c-kit, CD34, smooth muscle actin and desmin, with a Ki-67 index of <5%. Thus, the diagnosis in this case was benign schwannoma of the ascending colon.
Beneficial Bacteria Affect Danio rerio Development by the Modulation of Maternal Factors Involved in Autophagic, Apoptotic and Dorsalizing Processes
Authors: Miccoli Andrea , Gioacchini Giorgia , Maradonna Francesca , Benato Francesca , Skobo Tatjana , Carnevali Oliana
Published: June 2015

Background/Aims: Probiotic strains have been recognized to exert important roles in many biological systems, including immune response, growth, development and reproduction. However, to date, no studies have focused either on the relation among probiotics and maternal factors or on probiotics' ability to qualitatively and/or quantitatively modulate maternal transcripts. Methods: In this study, the effects of Lactobacillusrhamnosus administered to parental fish on the control of maternal factors involved in autophagic, apoptotic and dorsalizing processes during zebrafish embryo development were assessed through q-PCRs, WMISH and TUNEL assay. Results: The results we obtained show that probiotic induced significant changes in both maternal and zygotic mRNA levels involved in embryo development. The maternal autophagy-regulating genes herein investigated -ambra1a, ambra1b, beclin, lc3-, as well as those involved in the apoptotic process -caspase3, bcl2, bax- were modulated in disfavor and favor of the treated group, respectively. Also, the key transcripts ruling the dorsalizing process -goosecoid and chordin- were subject to a significant regulation of their gene expression. Conclusion: The results we acquired demonstrated that parentally administered Lactobacillusrhamnosus is able to modulate important physiological processes involved in zebrafish embryo development.
Stem Cell Conditioned Culture Media Attenuated Albumin-Induced Epithelial-Mesenchymal Transition in Renal Tubular Cells
Authors: Hu Junping , Zhu Qing , Li Pin-Lan , Wang Weili , Yi Fan , Li Ningjun
Published: June 2015

Background: Proteinuria-induced epithelial-mesenchymal transition (EMT) plays an important role in progressive renal tubulointerstitial fibrosis in chronic renal disease. Stem cell therapy has been used for different diseases. Stem cell conditioned culture media (SCM) exhibits similar beneficial effects as stem cell therapy. The present study tested the hypothesis that SCM inhibits albumin-induced EMT in cultured renal tubular cells. Methods: Rat renal tubular cells were treated with/without albumin (20 µmg/ml) plus SCM or control cell media (CCM). EMT markers and inflammatory factors were measured by Western blot and fluorescent images. Results: Albumin induced EMT as shown by significant decreases in levels of epithelial marker E-cadherin, increases in mesenchymal markers fibroblast-specific protein 1 and a-smooth muscle actin, and elevations in collagen I. SCM inhibited all these changes. Meanwhile, albumin induced NF-κB translocation from cytosol into nucleus and that SCM blocked the nuclear translocation of NF-κB. Albumin also increased the levels of pro-inflammatory factor monocyte chemoattractant protein-1 (MCP)-1 by nearly 30 fold compared with control. SCM almost abolished albumin-induced increase of MCP-1. Conclusion: These results suggest that SCM attenuated albumin-induced EMT in renal tubular cells via inhibiting activation of inflammatory factors, which may serve as a new therapeutic approach for chronic kidney diseases.
Nuclear SMAD2 Restrains Proliferation of Glioblastoma
Authors: Yu Yunhu , Ran Qishan
Published: June 2015

Aims: Although TGFβ receptor signaling has been shown to play a role in regulation of the growth and metastasis of glioblastoma multiforme (GBM), the downstream pathway through either SMAD2 or SMAD3 has not been elucidated. In this study, we investigate whether nuclear SMAD2 can restrain the proliferation of glioblastoma. Methods: A total of 23 resected specimens from GBM patients were collected for SMAD2 detection. Human GBM cell line A172, U87mg, D341m and Hs683 were maintained in Dulbecco's modified Eagle's medium and transfected with SMAD2 and SMAD3 shRNA plasmids. Gene expression was detected by RT-qPCR and Western and cell growth were detected by MTT assay. Results: Our results showed that the phosphorylated SMAD2 (pSMAD2, the nuclear and functional form of SMAD2) levels in GBM were significantly lower than the paired normal brain tissue in patients. Depletion of SMAD2, but not SMAD3, significantly abolished the inhibitory effects of TGFβ1 on the growth of GBM cells, possibly through pSMAD2-mediated increases in cell-cycle inhibitor, p27. Conclusion: Our data suggest that TGFβ/SMAD2 signaling cascades restrains growth of GBM.
Enhanced Epithelial-to-Mesenchymal Transition Associated with Lysosome Dysfunction in Podocytes: Role of p62/Sequestosome 1 as a Signaling Hub
Authors: Li Guangbi , Li Cai-Xia , Xia Min , Ritter Joseph K , Gehr Todd W. B. , Boini Krishna , Li Pin-Lan
Published: June 2015

Background: Autophagy is of importance in the regulation of cell differentiation and senescence in podocytes. It is possible that derangement of autophagy under different pathological conditions activates or enhances Epithelial-to-Mesenchymal Transition (EMT) in podocytes, resulting in glomerular sclerosis. To test this hypothesis, the present study produced lysosome dysfunction by inhibition of the vacuolar H+-ATPase (V-ATPase) to test whether deficiency of autophagic flux leads to enhancement of EMT in podocytes. Methods and Results: By Western blot and confocal analysis, lysosome inhibition using a V-ATPase inhibitor or its siRNA was found to markedly decreases the epithelial markers (P-cadherin and ZO-1) and increases the mesenchymal markers (FSP-1 and α-SMA). This enhancement was accompanied by deficient autophagic flux, as demonstrated by marked increases in LC3B-II and p62/Sequestosome 1. However, inhibition of autophagosome formation using spaudin-1 significantly attenuated both enhancement of EMT and deficiency of autophagic flux. To explore the mechanisms by which deficient autophagic flux enhances EMT, we tested the role of accumulated p62 as a signal hub in this process. Neither the nuclear factor erythroid 2-related factor 2 (Nrf2) and nuclear kappa-light-chain-enhancer pathways of p62 contributed to enhanced EMT. However, inhibition of cyclin-dependent kinase 1 (CDK1) activity reduced the phosphorylation of p62 and enhanced EMT in podocytes similar to lysosome dysfunction. Conclusion: The lack of phosphorylated p62 leads to a faster exit from cell mitosis, enhanced EMT associated with lysosome dysfunction may be attributed to accumulation of p62 and associated reduction of p62 phosphorylation.
CTGF Positively Regulates BMP12 Induced Tenogenic Differentiation of Tendon Stem Cells and Signaling
Authors: Liu Junpeng , Tao Xu , Chen Lei , Han Weifeng , Zhou You , Tang Kanglai
Published: June 2015

Background/Aims: Disordered differentiation of tendon stem cells (TSCs) during repair of injured tendon can result in the pathogenesis of chronic tendinopathy. Understanding tenocyte differentiation may provide new therapeutic insights for the prevention and treatment of chronic tendinopathy. The aim of our study was to determine if CTGF exerts a similar effect on BMP12-driven differentiation of rat TSCs. Methods: In overexpressing and RNA interference CTGF TSCs, tenogenic differentitation and the expression of related genes were determined by immunofluorescence staining, quantitative PCR, and western blotting, with or without BMP12 treatment. The interaction in vitro between CTGF and BMP12 was detected by Chemical crosslinking assay. Results: Our results showed that BMP12 effectively increased the expression of the tenocyte lineage markers scleraxis (Scx) and tenomodulin (Tnmd) at both mRNA and protein levels. Over-expression of CTGF from a lentiviral vector increased the expression of Scx and Tnmd as well as tendon proteins type I collagen (ColI) and tenascin-C (Tn-C) in TSCs compared to non-treated control cells with or without simultaneous BMP12 stimulation. Knockdown of CTGF expression decreased the expression of Scx, Tnmd, ColI and Tn-C compared to control cells. Chemical crosslinking experiments demonstrated a direct interaction between CTGF and BMP12. Conclusion: In conclusion, BMP12 plays a crucial role in tenogenesis via the Smad1/5/8 pathway, and CTGF positively promotes this effect.
Cloning and Expression of a Novel Target Fusion Protein and its Application in Anti-Tumor Therapy
Authors: Sun Rui , Zhu Yan , Feng Hailiang , Yang Zhenli , Bian Xiaocui , Gu Pei , Wang Chunjing , Liu Yuqin
Published: June 2015

Backgrounds: Epidermal growth factor (EGF) is a 53 amino acid polypeptide and its receptor EGFR is an established therapeutic target for anti-tumor therapy. Two major categories of EGFR-targeted drugs include monoclonal antibodies (mAbs) and small molecular tyrosine kinase inhibitors (TKIs). However, drug resistance occurs in a significant proportion of patients due to EGFR mutations. Since EGFR can maintain activation while abrogating the activity of mAbs or TKIs, or bypass signaling functions while successfully circumventing the EGF-EGFR switch, developing new mechanism-based inhibitors is necessary. Methods: In this study, based on the principle of tumor immunotherapy, a recombinant protein pLLO-hEGF was constructed. The N-terminal portion contains three immunodominant epitopes from listeriolysin O (LLO) and the C-terminal includes EGF. To use EGF as a target vector to recognize EGFR-expressing cancer cells, immunodominant epitopes could enhance immunogenicity of tumor cells for immune cell activation and attack. Results: Recombinant protein pLLO-hEGF was successfully expressed and showed strong affinity to cancer cells. Also, pLLO-hEGF could significantly stimulate human lymphocyte proliferation and the lymphocytes demonstrated enhanced killing potency in EGFR-expressing cancer cells in vitro and in vivo. Conclusion: This study can provide novel strategies and directions in tumor biotherapy.
Aberrant Expression of miRNA and mRNAs in Lesioned Tissues of Graves' Disease
Authors: Qin Qiu , Wang Xuan , Yan Ni , Song Rong-Hua , Cai Tian-Tian , Zhang Wen , Guan Li-Juan , Muhali Fatuma-Said , Zhang Jin-An
Published: June 2015

Background and Aims: Abnormal microRNA (miRNA) expression is found in many diseases including autoimmune diseases. However, little is known about the role of miRNA regulation in Graves' disease (GD). Here, we simultaneously detected different expressions of miRNA and mRNAs in thyroid tissues via a high-throughput transcriptomics approach, known as microarray, in order to reveal the relationship between aberrant expression of miRNAs and mRNAs spectrum and GD. Methods: Totally 7 specimens of thyroid tissue from 4 GD patients and 3 controls were obtained by surgery for microarray analysis. Then, 30 thyroid specimens (18 GD and 12 controls) were also collected for further validation by quantitative real-time PCR ( qRT-PCR ). Results: Statistical analysis showed that the expressions of 5 specific miRNA were increased significantly while those of other 18 miRNA were decreased in thyroid tissue of GD patients (FC≥1.3 or≤0.77 and p<0.05). In addition, the transcription of 1271 mRNAs was up-regulated, while the expression of 777 mRNAs transcripts was down-regulated (FC≥2.0 or≤0.5 and p<0.05). Furthermore, integrated analysis of differentially expressed miRNA and their target mRNAs demonstrated that 2 miRNA (miR-22 and miR-183) were increased while their potential target mRNAs were decreased. 3 miRNA (miR-101, miR-197 and miR-660) were decreased while their potential target mRNAs were increased. The above findings from microarray screening were confirmed by qRT-PCR in more samples. The results were consistent with those observed in the microarray assays. Conclusion: Our study highlights the possibility that miRNA-target gene network may be involved in the pathogenesis of GD and could provide new insights into understanding the pathophysiological mechanisms of GD.
Cardiac-Specific PID1 Overexpression Enhances Pressure Overload-Induced Cardiac Hypertrophy in Mice
Authors: Liu Yaoqiu , Shen Yahui , Zhu Jingai , Liu Ming , Li Xing , Chen Yumei , Kong Xiangqing , Song Guixian , Qian Lingmei
Published: June 2015

Background/Aims: PID1 was originally described as an insulin sensitivity relevance protein, which is also highly expressed in heart tissue. However, its function in the heart is still to be elucidated. Thus this study aimed to investigate the role of PID1 in the heart in response to hypertrophic stimuli. Methods: Samples of human failing hearts from the left ventricles of dilated cardiomyopathy (DCM) patients undergoing heart transplants were collected. Transgenic mice with cardiomyocyte-specific overexpression of PID1 were generated, and cardiac hypertrophy was induced by transverse aortic constriction (TAC). The extent of cardiac hypertrophy was evaluated by echocardiography as well as pathological and molecular analyses of heart samples. Results: A significant increase in PID1 expression was observed in failing human hearts and TAC-treated wild-type mouse hearts. When compared with TAC-treated wild-type mouse hearts, PID1-TG mouse showed a significant exacerbation of cardiac hypertrophy, fibrosis, and dysfunction. Further analysis of the signaling pathway in vivo suggested that these adverse effects of PID1 were associated with the inhibition of AKT, and activation of MAPK pathway. Conclusion: Under pathological conditions, over-expression of PID1 promotes cardiac hypertrophy by regulating the Akt and MAPK pathway.
Cost Analysis of Early versus Delayed Loop Ileostomy Closure: A Case-Matched Study
Authors: Robertson Jason , Linkhorn Hannah , Vather Ryash , Jaung Rebekah , Bissett Ian P.
Published: June 2015

Background/Aims: The optimal timing for the closure of loop ileostomies remains controversial. The aim of the current study was to investigate whether early ileostomy closure (EC) (<2 weeks post-formation) results in significant healthcare savings as against late closure (LC). Methods: Patients with available cost data that underwent EC between January 2008 and December 2012 were compared against matched patients undergoing LC during the same period. Direct hospital costs for the two groups were compared. Results: There were 42 EC patients and 61 LC patients. EC patients had significantly less ileostomy-related complications (p < 0.001) and hospital readmissions (p < 0.001). Operative time (p < 0.001) and operative cost (p = 0.002) were also both significantly less in the EC group. Community nursing costs favoured the LC group (p = 0.047). The EC group had an increased post-closure wound infection rate (p = 0.02). The mean total direct cost per patient was NZD 13,724 (SD NZD 3,736) for EC and NZD 16,728 (SD NZD 8,028) for LC. Representing an average costs saving of NZD 3,004 per patient favouring EC (p = 0.012). Conclusion: Although EC increases the post-closure wound infection rate, EC reduces ileostomy complications, hospital readmissions and operative costs resulting in significant healthcare savings. In order to improve patient outcomes and make EC even more cost effective, efforts should be taken to reduce wound infections.
Do Oral Health Conditions Adversely Impact Young Adults?
Authors: Carvalho Joana C. , Mestrinho Heliana D. , Stevens Sophie , van Wijk Arjen J.
Published: June 2015

This study assessed the extent to which clinically measured oral health conditions, adjusted for sociodemographic and oral health behavior determinants, impact adversely on the oral health-related quality of life (OHRQoL) in a sample of Belgian young adults. The null hypothesis was that, among young adults, the oral health conditions would have no impact on their quality of life. The participants were 611 new patients aged 16-32 years seeking consultation at the Saint-Luc University Hospital in Brussels in 2010-2011. The patients (56.0% female) were examined for their oral health conditions and answered a validated questionnaire about sociodemographic and oral health behavior determinants in addition to questions about their OHRQoL. The abridged Oral Health Impact Profile-14 was used to assess the OHRQoL. Interexaminer reliability for caries was 0.86 (95% CI 0.84-0.89, nonweighted κ). The outcome was a high score on the OHRQoL (median split). Hierarchical logistic regression analysis showed that young adults with clinical absolute D1MFS scores between 9 and 16 (OR = 2.14, p = 0.031) and between 17 and 24 (OR = 3.10, p = 0.003) were significantly more likely to report a high impact on their quality of life than those with lower scores. Also, periodontal conditions compromised significantly (OR = 1.79, p = 0.011) the quality of life of young adults. In conclusion, this study identified oral health conditions with a significant adverse effect on the OHRQoL of young adults. However, the prevalence of young adults reporting impacts on at least 1 performance affected fairly often or very often was limited to 18.7% of the sample.
Epidemiologie der aktinischen Keratose
Authors: Green Adèle C.
Published: June 2015

Die Epidemiologie der aktinischen Keratose (AK) spiegelt deren Verursachung durch kumulative Sonnenexposition wider: Am höchsten ist die Prävalenz bei hellhäutigen Menschen, die in niedrigen Breitengraden leben; besonders betroffen sind die Körperregionen, die der Sonne am meisten ausgesetzt sind, nämlich Hände, Unterarme und Gesicht. AK-Läsionen deuten auf ein erhöhtes Risiko für Basalzellkarzinom, Plattenepithelkarzinom und Melanom hin, vor allem wenn sie in großer Zahl vorliegen und sich zu Arealen sogenannter Feldkanzerisierung vereinigen. Die wichtigsten Risikofaktoren sind männliches Geschlecht, höheres Alter, sonnenempfindlicher Teint, hohe kumulative Sonnenexposition und längerfristige Immunsuppression. Die klinische Auszählung der AK ermöglicht die Beurteilung und Beobachtung der AK-Krankheitslast, jedoch ist eine wirklich präzise Zählung kaum möglich, besonders bei stark lichtgeschädigter Haut. Mehrfach ist nachgewiesen worden, dass AK-Auszählungen unzuverlässig sind, auch wenn sie durch spezialisierte Dermatologen vorgenommen werden. Ungeachtet dieser Herausforderungen zeigt die qualitative Betrachtung von AK-Verläufen eine hohe Umsatzrate, bei der neue Läsionen entstehen und sich vorhandene Läsionen zurückbilden. Ein sehr geringer Anteil der AK-Läsionen durchläuft eine maligne Transformation, jedoch ist die genaue Rate aufgrund der Mängel in der langfristigen AK-Beobachtung unbekannt. Die Primärprävention von AK besteht in der Meidung intensiver Sonneneinstrahlung durch Schutzmaßnahmen wie dem Aufenthalt im tiefen Schatten, schützender Kleidung und regelmäßigem Auftragen von Sonnenschutzmittel auf exponierte Hautareale von frühester Jugend an.
Lichtbedingte Schädigungen: Vorboten von aktinischer Keratose und frühem Plattenepithelkarzinom
Authors: Wei Jerry , Kok Lai Fong , Byrne Scott N. , Halliday Gary M.
Published: June 2015

Ultraviolette (UV-)Strahlung ist aller Wahrscheinlichkeit nach ein treibender Faktor für die Entstehung von Hauttumoren und die Progression von aktinischer Keratose zum Plattenepithelkarzinom. Lichtbedingte Schädigungen treten auf vielen verschiedenen Ebenen auf. UV-Strahlen verursachen Schäden an zahlreichen Zellbestandteilen - darunter Lipide, Proteine und DNA -, die wahrscheinlich allesamt zu UV-bedingtem Hautkrebs beitragen. Zwei biologische Ereignisse, die letztlich durch eine solche Lichtschädigung hervorgerufen werden, sind Mutationen in den Genen, die für die Steuerung der Zellteilung, -differenzierung und -wanderung verantwortlich sind, sowie Immunsuppression. Wenn lichtbedingte DNA-Schäden vor der Zellteilung nicht repariert werden, kann dies zum Einbau eines fehlerhaften Nukleotids in neu synthetisierte DNA führen. Mutationen in kritischen Genen tragen zur Karzinogenese bei. Eine lichtbedingte Schädigung von Proteinen, die z.B. an der DNA-Reparatur beteiligt sind, oder von Proteinen oder Lipiden, die an zellulärer Signalübermittlung partizipieren, kann diese Reparaturmechanismen behindern und zur Mutagenese beitragen. Mutationen in Schlüsselgenen wie TP53, BRM, PTCH1 oder HRAS können zur kutanen Karzinogenese beitragen. Durch UV-Strahlung wird auch die Immunabwehr beeinträchtigt. Lichtbedingte Schädigungen von DNA und Signallipiden sowie andere molekulare Veränderungen sind schädlich für wichtige Zellen der Immunregulation. Lichtgeschädigte dendritische Zellen und Mastzellen mit verändertem Verhalten führen zur Aktivierung von regulatorischen T- und B-Zellen; diese unterdrücken die Immunreaktion auf die Proteinprodukte UV-mutierter Gene. Dadurch kann das Immunsystem seine Schutzfunktion, mutierte Zellen zu zerstören, nicht mehr ausüben, und die transformierten Zellen können zu Hautkrebs progredieren. UV-Strahlung scheint bei jedem dieser Schritte eine zentrale Rolle zu spielen, daher sind Anzeichen lichtbedingter Schädigungen als Vorboten der Entstehung von Hautkrebs aufzufassen.
Three-Year Course of Cannabis Dependence and Prediction of Persistence
Authors: van der Pol Peggy , Liebregts Nienke , de Graaf Ron , Korf Dirk J. , van den Brink Wim , van Laar Margriet
Published: June 2015

Aims: To examine the course and the predictors of the persistence of cannabis dependence. Methods: Through cannabis outlets and chain referral, a prospective enriched community cohort of 207 young adults (aged 18-30) with DSM-IV cannabis dependence at baseline (T0) was formed and followed-up after 1.5 (T1) and 3 (T2) years. The presence of cannabis dependence, cannabis-related problems, functional impairment and treatment was assessed using the Composite International Diagnostic Interview (CIDI 3.0) and the Sheehan Disability Scale (SDS). Predictors of persistence were lifetime cannabis abuse and dependence symptoms, cannabis use characteristics, distant vulnerability factors (e.g. childhood adversity, family history of psychological/substance use problems, impulsivity, mental disorders), and proximal stress factors (recent life events, social support). Results: Four groups were distinguished: persistent dependent (DDD: 28.0%), stable non-persistent (DNN: 40.6%), late non-persistent (DDN: 17.9%) and recurrent dependent (DND: 13.5%). At T2, persisters (DDD) reported significantly more (heavy) cannabis use and cannabis problems than non-persisters (DNN/DDN/DND). Treatment seeking for cannabis-related problems was rare, even among persisters (15.5%). The number (OR = 1.23 (1.03-1.48)) and type (‘role impairment' OR = 2.85 (1.11-7.31), ‘use despite problems' OR = 2.34 (1.15-4.76)) of lifetime cannabis abuse/dependence symptoms were the only independent predictors of persistence with a total explained variance of 8.8%. Conclusions: Persistence of cannabis dependence in the community is low, difficult to predict, and associated with a negative outcome. The substantial proportion of stable non-persisters suggests that screening and monitoring or low-threshold brief interventions may suffice for many non-treatment-seeking cannabis-dependent people. However, those with many lifetime abuse/dependence symptoms may benefit from more intensive interventions.
Lethality of Opioid Overdose in a Community Cohort of Young Heroin Users
Authors: Espelt Albert , Barrio Gregorio , Álamo-Junquera Dolores , Bravo Maria José , Sarasa-Renedo Ana , Vallejo Fernando , Molist Gemma , Brugal M. Teresa ,   ITINERE Project Group
Published: June 2015

Background: The aim of the study was to estimate the lethality of opioid overdose among young heroin users. Methods: A prospective community cohort study was conducted in Barcelona and Madrid, Spain. Participants included 791 heroin users aged 18-30 years who were followed up between 2001 and 2006. Fatal overdoses were identified by record linkage of the cohort with the general mortality register, while non-fatal overdoses were self-reported at baseline and follow-up interviews. The person-years (py) at risk were computed for each participant. Fatal and non-fatal overdose rates were estimated by city. Transition towards injection shortly before the overdose could not be measured. Overdose lethality (rate of fatal overdose in proportion to total overdose) and its 95% CI was estimated using Bayesian models. Results: The adjusted rates of fatal and non-fatal opioid overdose were 0.7/100 py (95% CI: 0.4-1.1) and 15.8/100 py (95% CI: 14.3-17.6), respectively. The adjusted lethality was 4.2% (95% CI: 2.5-6.5). Conclusions: Four out of 100 opioid overdoses are fatal. These are preventable deaths that could be avoided before or after the overdose takes place. Resources are urgently needed to prevent fatal opioid overdose.
Neuropathic Pain, Depression, and Cardiovascular Disease: A National Multicenter Study
Authors: Cragg Jacquelyn J. , Noonan Vanessa K. , Noreau Luc , Borisoff Jaimie F. , Kramer John K.
Published: June 2015

Background: Individuals with spinal cord injury (SCI) have a more than twofold increased risk of heart disease and stroke compared with able-bodied individuals. The increased risk appears to be in excess of the risk conferred by several well-established risk factors, including diabetes, hypertension, and sex. This raises the question whether other factors, secondary to SCI, are also contributing to the development of cardiovascular disease (CVD). Two potential factors associated with SCI and CVD are pain and depression. Both are frequently reported among individuals with SCI, develop in the acute stages of injury, and are commonly described as severe. Therefore, the primary aim of this study was to examine the relationship between pain (and types of pain) and depression with CVD among individuals with SCI. Methods: A total of 1,493 individuals (referred sample) with chronic SCI participated in a self-report cross-sectional multicenter Canada-wide survey from 2011-2012 (mean age ± standard deviation: 49.6 ± 13.9 years). Results: After adjustment for age, sex, and injury characteristics, neuropathic pain and depression were significantly and independently associated with CVD (adjusted odds ratio and 95% confidence interval: 2.27 (1.21, 4.60) for neuropathic pain; 4.07 (2.10, 7.87) for depression). In contrast to neuropathic pain, non-neuropathic pain was not significantly associated with CVD (p = 0.13). Conclusion: In conclusion, these data illustrate important interrelationships between secondary complications following SCI, as well as raise the possibility of neuropathic pain (versus nociceptive pain) as a novel and emerging risk factor for CVD.