bluuebunny/crossref_metadata_2025 huggingface dataset:
Randomized study of low‐dose versus standard‐dose chemoradiotherapy for unresectable esophageal squamous cell carcinoma (JCOG0303)
Authors: Shinoda Masayuki , Ando Nobutoshi , Kato Ken , Ishikura Satoshi , Kato Hoichi , Tsubosa Yasuhiro , Minashi Keiko , Okabe Hiroshi , Kimura Yusuke , Kawano Tatsuyuki , Kosugi Shin‐Ichi , Toh Yasushi , Nakamura Kenichi , Fukuda Haruhiko ,   Japan Clinical Oncology Group
Published: January 2015

Low‐dose cisplatin and 5‐fluorouracil (LDPF) chemotherapy with daily radiotherapy (RT) is used as an alternative chemoradiotherapy regimen for locally advanced esophageal carcinoma. We evaluated whether RT plus LDPF chemotherapy had an advantage in terms of survival and/or toxicity over RT plus standard‐dose cisplatin and 5‐fluorouracil (SDPF) chemotherapy in this study. This multicenter trial included esophageal cancer patients with clinical T4 disease and/or unresectable regional lymph node metastasis. Patients were randomly assigned to receive RT (2 Gy/fraction, total dose of 60 Gy) with SDPF (arm A) or LDPF (arm B) chemotherapy. The primary endpoint was overall survival (OS). A total of 142 patients (arm A/B, 71/71) from 41 institutions were enrolled between April 2004 and September 2009. The OS hazard ratio in arm B versus arm A was 1.05 (80% confidence interval, 0.78–1.41). There were no differences in toxicities in either arm. Arm B was judged as not promising for further evaluation in the phase III setting. Thus, the Data and Safety Monitoring Committee recommended that the study be terminated. In the updated analyses, median OS and 3‐year OS were 13.1 months and 25.9%, respectively, for arm A and 14.4 months and 25.7%, respectively, for arm B. Daily RT plus LDPF chemotherapy did not qualify for further evaluation as a new treatment option for patients with locally advanced unresectable esophageal cancer. This study was registered at the UMIN Clinical Trials Registry as UMIN000000861.
New metastatic model of human small‐cell lung cancer by orthotopic transplantation in mice
Authors: Sakamoto Shuichi , Inoue Hiroyuki , Ohba Shunichi , Kohda Yasuko , Usami Ihomi , Masuda Tohru , Kawada Manabu , Nomoto Akio
Published: January 2015

Small‐cell lung cancer (SCLC) is an aggressive cancer with high metastatic ability and novel strategies against the metastasis are urgently needed to improve SCLC treatment. However, the mechanism of metastasis of SCLC remains largely to be elucidated. For further studies of SCLC metastasis, we developed a new orthotopic transplantation model in mice. We established a GFP‐labeled subline from the human SCLC cell line DMS273 and transplanted them orthotopically into the lung of nude mice with Matrigel. The GFP‐labeled cells showed significant metastatic activity and formed metastatic foci in distant tissues such as bone, kidney, and brain, as observed in SCLC patients. From a bone metastasis focus of the mouse, we isolated another subline, termed G3H, with enhanced metastatic potential and higher hepatocyte growth factor (HGF) expression than the parental line. Further studies indicated that the HGF/MET signaling pathway was involved in in vitro motility and invasion activities of the G3H cells and treatments with MET inhibitors decreased formation of distant metastases in our orthotopic model using G3H cells. These data indicated that our model mimics the clinical aspect of SCLC such as metastatic tropism and autocrine of HGF/MET signaling. Compared with other orthotopic SCLC models, our model has a superior ability to form distant metastases. Therefore, our model will provide a valuable tool for the study of SCLC metastasis.
Parental and family‐related influences on dental caries in children of Dutch, Moroccan and Turkish origin
Authors: Duijster Denise , de Jong‐Lenters Maddelon , de Ruiter Corine , Thijssen Jill , van Loveren Cor  , Verrips Erik
Published: January 2015

Objectives

The aim of this cross‐sectional study was to investigate the relationship between parental and family‐related factors and childhood dental caries in a sample of 5‐ to 6‐year‐old children of Dutch, Moroccan and Turkish origin. Furthermore, the relationship of parental and family‐related factors with social class and ethnicity was examined.

Methods

The study sample included 92 parent–child dyads (46 cases and 46 controls), which were recruited from a large paediatric dental centre in The Hague, the Netherlands. Cases were children with four or more decayed, missing or filled teeth, and controls were caries free. Validated questionnaires were used to collect data on sociodemographic characteristics, oral health behaviours, parents’ dental self‐efficacy and locus of control (LoC), parenting practices and family functioning. Parenting practices were also assessed using structured video observations of parent–child interactions.

Results

Parents of controls had a more internal LoC, and they were more likely to show positive (observed) parenting in terms of positive involvement, encouragement and problem‐solving, compared to cases (P < 0.05). Lower social class was significantly associated with a lower dental self‐efficacy, a more external LoC and poorer parenting practices. Furthermore, LoC was more external in Moroccan and Turkish parents, compared to Dutch parents.

Conclusion

Parents’ internal LoC and observed positive parenting practices on the dimensions positive involvement, encouragement and problem‐solving were important indicators of dental health in children of Dutch, Moroccan and Turkish origin. Findings suggest that these parental factors are potential mediators of socioeconomic inequalities in children's dental health.
Activated natural killer cells accelerate liver damage in patients with chronic hepatitis B virus infection
Authors: Zheng Q , Zhu Y Y , Chen J , Ye Y B , Li J Y , Liu Y R , Hu M L , Zheng Y C , Jiang J J
Published: January 2015

Emerging evidence indicates that natural killer (NK) cells may contribute to liver injury in patients with hepatitis B virus (HBV) infection. Because HBV infection progresses through various disease phases, the cytolytic profiles of peripheral and intrahepatic NK cells in HBV-infected patients remain to be defined. In this study, we comprehensively characterized intrahepatic and peripheral NK cells in a cohort of HBV-infected individuals, and investigated their impact on liver pathogenesis during chronic HBV infection. The study population included 34 immune-clearance (IC) patients, 36 immune-tolerant (IT) carriers and 10 healthy subjects. We found that the activity of peripheral NK cells from IC patients was functionally elevated compared to IT carriers and controls, and NK cell activation was indicated by an increased expression of CD69, CD107a, interferon (IFN)-γ and tumour necrosis factor (TNF)-α. Further analysis showed that the increased activity of both peripheral and hepatic NK cells was correlated positively with liver injury, which was assessed by serum alanine aminotransferase levels (ALT) and the liver histological activity index (HAI). Interestingly, the frequency of peripheral NK cells was reduced in IC patients (especially those with higher HAI scores of 3–4), but there was a concomitant increase in hepatic NK cells. The functionally activated NK cells are enriched preferentially in the livers of IC patients and skew towards cytolytic activity that accelerates liver injury in chronic hepatitis B (CHB) patients.
408 kb 15q11.2 microduplication by array comparative genomic hybridization in a fetus presenting with exomphalos, micrognathia, tetralogy ofFallot and normal karyotype: A genetic counseling dilemma in paternal carrier status
Authors: Tonni Gabriele , Bellotti Maria , Palmisano Marcella , Alesi Viola , Bertoli Marta , Bonasoni Maria Paola
Published: January 2015

Exomphalos may be associated with chromosomal abnormalities and syndromes. Severe exomphalos (herniation of liver, midgut and spleen) associated with increased nuchal translucency was seen at first trimester screening test. Karyotype by chorionic villus sampling showed normal male fetus. Follow up scan at 16 and 18 weeks of gestation confirmed the severe exomphalos and detected micrognathia and tetralogy ofFallot. Array comparative genomic hybridization (a‐CGH) further demonstrated a 408 kb 15q11.2 microduplication, with the father‐to‐be as healthy carrier. This is the first case of an association between 15q11.2 micorduplication and fetal sonographic anomalies. Genetic counseling for estimation of recurrent risk of congenital anomalies is discussed.
Problems with supertrees based on the subtree prune‐and‐regraft distance, with comments on majority rule supertrees
Authors: Goloboff Pablo A. , Szumik Claudia A.
Published: January 2015

This paper examines a recent proposal to calculate supertrees by minimizing the sum of subtree prune‐and‐regraft distances to the input trees. The supertrees thus calculated may display groups present in a minority of the input trees but contradicted by the majority, or groups that are not supported by any input tree or combination of input trees. The proponents of the method themselves stated that these are serious problems of “matrix representation with parsimony”, but they can in fact occur in their own method. The majority rule supertrees, being explicitly clade‐based, cannot have these problems, and seem much more suited to retrieving common clades from a set of trees with different taxon sets. However, it is dubious that so‐called majority rule supertrees can always be interpreted as displaying those clades present (or compatible with) with a majority of the trees. The majority rule consensus is always a median tree, in terms of the Robinson–Foulds distances (i.e. it minimizes the sum of Robinson–Foulds distances to the input trees). In contrast, majority rule supertrees may not be median—different, contradictory trees may minimize Robinson–Foulds distances, while their strict consensus does not. If being “majority” results from being median in Robinson–Foulds distances, this means that in the supertree setting a “majority” is ambiguously defined, sometimes achievable only by mutually contradictory trees.
THE IMPACT OF SODA SALES TAXES ON CONSUMPTION: EVIDENCE FROM SCANNER DATA
Authors: Colantuoni Francesca , Rojas Christian
Published: January 2015

Scientific evidence on the effect of sugar consumption on obesity has propelled policy makers in several states across the United States to propose the imposition of a tax on soft drinks sales. In this article, we look at the effect of two tax events: a 5.5% sales tax on soft drinks imposed by the state of Maine in 1991 and a 5% sales tax on soft drinks levied in Ohio in 2003. We investigate this question by using sales data collected by scanner devices in the two states, where soda taxes where enacted as well as on neighboring states. We employ a difference‐in‐difference matching estimator (DIDM) that, in our setting, permits the comparison among treatment and control groups based on brand identity. Results suggest that neither sales tax had a statistically significant impact on the consumption of soft drinks. This finding is robust to several alternative specifications. (JEL D12, H22, C90, L66)
Effect of transcutaneous electrical nerve stimulation on peripheral to central blood pressure ratio in healthy subjects
Authors: da Silva Marianne L. , Chiappa Gaspar R. , da Silva Vinicius M. , Neves Laura M. T. , de Lima Alexandra C. G. B. , Tomasi Fernanda P. , Junior Luiz T. G. , Vilela‐Martin José F. , Bottaro Martim , Junior Gerson C.
Published: January 2015

Purpose

To investigate the effect of transcutaneous electrical nerve stimulation (TENS) on the arterial stiffness in healthy young adult and middle‐aged men using the augmentation index (AI‐x) and hemodynamic measures.

Methods

Twenty‐four men (12 aged 27·25 ± 5·53 years and 12 aged 54·83 ± 5·10 years) were randomly allocated to two subgroups: TENS or placebo in ganglion region for 45 min. The AI‐x and hemodynamic measures [late systolic blood pressure (SBP), central blood pressure (CBP), difference between P1 and P2 (ΔP) and tension time index (TTI)] were determined before and after protocols.

Results

TENS resulted in reduction of SBP in younger adults (TENSpre: 111 ± 2; post: 105 ± 2·2 mm Hg; PLACEBOpre: 113 ± 1·8; post: 114 ± 2·5 mm Hg; GEE, P<0·01), whereas no difference was found in middle‐aged group. TENS also resulted in reduction of AI‐x younger adults group (TENSpre: 56 ± 2·8; post: 53 ± 2%; PLACEBOpre: 55 ± 3; post: 58 ± 2·5%; GEE, P<0·01). ΔP and TTI were significantly decreased after the application of TENS in both groups, but significantly greater reductions in TTI and the SBP/CBP ratio were found in the group of younger adults.

Conclusions

The acute application of ganglion TENS attenuated arterial stiffness in younger adults as well as hemodynamic measures in the middle‐aged group. This method could emerge as effective therapy for the management of arterial blood pressure.
Contrast‐enhanced harmonic endoscopic ultrasonography for pancreatobiliary diseases
Authors: Kitano Masayuki , Kamata Ken , Imai Hajime , Miyata Takeshi , Yasukawa Satoru , Yanagisawa Akio , Kudo Masatoshi
Published: January 2015

The combination of second‐generation ultrasound contrast agents and an endoscopic ultrasonography (EUS) system with a broad‐band transducer has allowed contrast‐enhanced harmonic imaging in the field of EUS. In contrast‐enhanced harmonic EUS (CH‐EUS), diffuse homogeneous enhancement is obtained in normal parenchyma of the pancreas. The bile duct and pancreatic duct are depicted as non‐enhanced ductal structures with strong contrast in comparison to the surrounding parenchyma. CH‐EUS identifies pancreatic adenocarcinomas as solid lesions exhibiting hypo‐enhancement with a sensitivity and specificity of 88–96% and 88–94%, respectively. In particular, 80–100% of false‐negative cases in endoscopic ultrasound‐guided fine‐needle aspiration (EUS‐FNA) are correctly classified by CH‐EUS, suggesting CH‐EUS complements EUS‐FNA. Moreover, CH‐EUS improves depiction of some subtle lesions in conventional EUS, thus facilitating EUS‐FNA. For quantitative perfusion analysis, a time–intensity curve (TIC) for the region of interest can be generated during CH‐EUS. The maximum intensity gain and the echo intensity reduction rate from the peak at 1 min obtained by TIC can be used for differentiation of pancreatic adenocarcinoma from other tumors. CH‐EUS is also useful for differentiation of invasive intraductal papillary mucinous neoplasms (IPMN) from non‐invasive IPMN, identification of malignant lesions in the gallbladder, and T‐ and N‐staging of pancreatobiliary tumors.
Phosphorylation of Smad2/3 at the specific linker threonine residue indicates slow-cycling esophageal stem-like cells before re-entry to the cell cycle
Authors: Takahashi Y. , Fukui T. , Kishimoto M. , Suzuki R. , Mitsuyama T. , Sumimoto K. , Okazaki T. , Sakao M. , Sakaguchi Y. , Yoshida K. , Uchida K. , Nishio A. , Matsuzaki K. , Okazaki K.
Published: January 2015

The stem cell compartment in the esophageal epithelium is possibly located in the basal layer. We have identified significant expression of Smad2/3, phosphorylated at specific linker threonine residues (pSmad2/3L-Thr), in the epithelial cells of murine stomach and intestine, and have suggested that these cells are epithelial stem cells. In this study, we explore whether pSmad2/3L-Thr could serve as a biomarker for esophageal stem cells. We examined esophageal tissues from normal C57BL/6 mice and those with esophagitis. Double immunofluorescent staining of pSmad2/3L-Thr with Ki67, CDK4, p63, or CK14 was performed. After immunofluorescent staining, we stained the same sections with hematoxylin-eosin and observed these cells under a light microscope. We used the 5-bromo-2-deoxyuridine (BrdU) labeling assay to examine label retention of pSmad2/3L-Thr immunostaining-positive cells. We collected specimens 5, 10, 15 and 20 days after repeated BrdU administrations and observed double immunofluorescent staining of pSmad2/3L-Thr with BrdU. In the esophagus, pSmad2/3L-Thr immunostaining-positive cells were detected in the basal layer. These cells were detected between Ki67 immunostaining-positive cells, but they were not co-localized with Ki67. pSmad2/3L-Thr immunostaining-positive cells showed co-localization with CDK4, p63, and CK14. Under a light microscope, pSmad2/3L-Thr immunostaining-positive cells indicated undifferentiated morphological features. Until 20 days follow-up period, pSmad2/3L-Thr immunostaining-positive cells were co-localized with BrdU. pSmad2/3L-Thr immunostaining-positive cells significantly increased in the regeneration phase of esophagitis mucosae, as compared with control mice (esophagitis vs. control: 6.889 ± 0.676/cm vs. 4.293 ± 0.659/cm; P < 0.001). We have identified significant expression of pSmad2/3L-Thr in the specific epithelial cells of murine esophagi. We suggest that these cells are slow-cycling epithelial stem-like cells before re-entry to the cell cycle.
Effects of Frame Rate on Two‐Dimensional Speckle Tracking–Derived Measurements of Myocardial Deformation in Premature Infants
Authors: Sanchez Aura A. , Levy Philip T. , Sekarski Timothy J. , Hamvas Aaron , Holland Mark R. , Singh Gautam K.
Published: January 2015

Background

Frame rate (FR) of image acquisition is an important determinant of the reliability of 2‐dimensional speckle tracking echocardiography (2DSTE)‐derived myocardial strain. Premature infants have relatively high heart rates (HR). The aim was to analyze the effects of varying FR on the reproducibility of 2DSTE‐derived right ventricle (RV) and left ventricle (LV) longitudinal strain (LS) and strain rate (LSR) in premature infants.

Methods

RV and LV LS and LSR were measured by 2DSTE in the apical four‐chamber view in 20 premature infants (26 ± 1 weeks) with HR 163 ± 13 bpm. For each subject, 4 sets of cine loops were acquired at FR of <90, 90–110, 110–130, and >130 frames/sec. Two observers measured LS and LSR. Inter‐ and intra‐observer reproducibility was assessed using Bland–Altman analysis, coefficient of variation, and linear regression.

Results

Intra‐observer reproducibility for RV and LV LS was higher at FR >110 frames/sec, and optimum at FR >130 frames/sec. The highest inter‐observer reproducibility for RV and LV LS were at FR >130 and >110 frames/s, respectively. The highest reproducibility for RV and LV systolic and early diastolic LSR was at FR >110 frames/sec. FR/HR ratio >0.7 frames/sec per bpm yielded optimum reproducibility for RV and LV deformation imaging.

Conclusions

The reliability of 2DSTE‐derived RV and LV deformation imaging in premature infants is affected by the FR of image acquisition. Reproducibility is most robust when cine loops are obtained with FR/HR ratio between 0.7 and 0.9 frames/sec per bpm, which likely results from optimal myocardial speckle tracking and mechanical event timing.
Three‐Dimensional Echocardiography‐Based Prediction of Posterior Leaflet Resection
Authors: Rim Yonghoon , Choi Ahnryul , Laing Susan T. , McPherson David D. , Kim Hyunggun
Published: January 2015

Clinical long‐term outcomes have shown that partial leaflet resection followed by ring annuloplasty is a reliable and reproducible surgical repair technique for treatment of mitral valve (MV) leaflet prolapse. We report a 61‐year‐old male for three‐dimensional transesophageal echocardiography (3DTEE)‐based virtual posterior leaflet resection and ring annuloplasty. Severe mitral regurgitation was found and computational evaluation demonstrated substantial leaflet malcoaptation and high stress concentration. Following virtual resection and ring annuloplasty, posterior leaflet prolapse markedly decreased, sufficient leaflet coaptation was restored, and high stress concentration disappeared. Virtual MV repair strategies using 3DTEE have the potential to help optimize MV repair.
RISK ASSESSMENT UNDER A NONLINEAR FISCAL POLICY RULE
Authors: Shiamptanis Christos
Published: January 2015

In the aftermath of the recent debt crisis, many countries are implementing nonlinear fiscal policy rules, whereby the government's responsiveness to debt strengthens at higher levels of debt. This paper examines how a nonlinear fiscal policy rule affects the possibility of future insolvency in a small open economy. We find that (1) the criteria for a nonlinear fiscal rule to eliminate explosive behavior should be tighter than the ones proposed by Bohn (1998); (2) a country that adopts a nonlinear fiscal rule could substantially reduce the probability of a solvency crisis; and (3) a nonlinear fiscal rule allows a country to reduce the possibility of insolvency without large initial responsiveness. (JEL C63, E62, E63, F34, H63)
Individualistic response to past climate changes: niche differentiation promotes diverging Quaternary range dynamics in the subspecies of Testudo graeca
Authors: Anadón J. D. , Graciá E. , Botella F. , Giménez A. , Fahd S. , Fritz U.
Published: January 2015

Understanding how species have responded to past climate changes might inform how current climate change may affect species. However, current perspectives on species’ response to Quaternary cycles mainly stem from studies on mid and high latitudes and limited information is available at lower latitudes. In these systems a greater variation in individual species’ responses to glacial cycles has been suggested, but we lack of clear case studies of such variations. Here, by means of ecological niche modelling, we characterized the niche differentiation of the five subspecies of the tortoise Testudo graeca in north Africa and described the range dynamics suffered by each of the subspecies since the Last Glacial Maxima. Molecular data was employed to infer past population dynamics.

The five subspecies present a clear niche differentiation, particularly in relation to rainfall, covering a range from semiarid to humid species. The ecological differentiation among subspecies promotes very different, even opposite, post‐glacial range dynamics. Since the Last Glacial Maxima, the ranges have either expanded (T. g. graeca), contracted (T. g. marokkensis), shifted northwards (T. g. soussensis) or remained stable (T. g. cyrenaica and T. g. nabeulensis). Molecular data supported the majority of these range dynamics. Our work exemplifies how, in a climate change scenario, phylogenetically very close taxa (i.e. at the subspecies level) might experience strikingly different biogeographical dynamics due to former niche differentiation. Our work supports an individualistic response to glacial cycles, which can be particularly strong in lower latitudes. In these areas precipitation changes during glacial periods may have strongly affected distribution ranges in idiosyncratic ways. We hypothesize that in those species limited by precipitation the response to glacial cycles (and climate changes) is more unpredictable than in those limited by temperature.
Seasonal decline in plant defence is associated with relaxed offensive oviposition behaviour in the viburnum leaf beetle Pyrrhalta viburni
Authors: DESURMONT GAYLORD A. , HAJEK ANN E. , AGRAWAL ANURAG A.
Published: January 2015

1. Plant defence of Viburnum shrubs against oviposition by its specialist herbivore, the viburnum leaf beetle [VLB Pyrrhalta viburni(Paykull)], involves an egg‐crushing wound response in twigs. Although the response is variable among Viburnum species, it can have a strong impact on egg survivorship. Beetles typically aggregate egg masses with conspecifics along infested twigs, forming clusters that can overwhelm the twig response. It was investigated whether twig responses and beetle oviposition behaviour vary seasonally.

2. In a field experiment, twig defences decreased towards the end of the VLB oviposition period: wound response of the North American Viburnum dentatum L. and the European V. opulus L. was reduced by 100% and 54%, respectively, in September compared with the July to August period.

3. Oviposition trials demonstrated a corresponding behavioural change: VLB females displayed aggregative oviposition in August, but not in September.

4. Further tests revealed that late‐season VLB females reverted to aggregative oviposition after being kept on uninfested twigs, whereas females kept on heavily infested twigs did not. This behavioural change suggests that relaxation of aggregative oviposition originates from cues associated with high densities of egg masses.

5. Relaxation of aggregative oviposition may be adaptive (and beneficial for invasion) on shrubs with low levels of defences by reducing intra‐specific competition.
Isolated and combined dystonia syndromes – an update on new genes and their phenotypes
Authors: Balint B. , Bhatia K. P.
Published: January 2015

Recent consensus on the definition, phenomenology and classification of dystonia centres around phenomenology and guides our diagnostic approach for the heterogeneous group of dystonias. Current terminology classifies conditions where dystonia is the sole motor feature (apart from tremor) as ‘isolated dystonia’, while ‘combined dystonia’ refers to dystonias with other accompanying movement disorders. This review highlights recent advances in the genetics of some isolated and combined dystonic syndromes. Some genes, such as ANO3, GNAL and CIZ1, have been discovered for isolated dystonia, but they are probably not a common cause of classic cervical dystonia. Conversely, the phenotype associated with TUBB4A mutations expanded from that of isolated dystonia to a syndrome of hypomyelination with atrophy of the basal ganglia and cerebellum (H‐ABC syndrome). Similarly, ATP1A3 mutations cause a wide phenotypic spectrum ranging from rapid‐onset dystonia‐parkinsonism to alternating hemiplegia of childhood. Other entities entailing dystonia‐parkinsonism include dopamine transporter deficiency syndrome (SLC63 mutations); dopa‐responsive dystonias; young‐onset parkinsonism (PARKIN, PINK1 and DJ‐1 mutations); PRKRA mutations; and X‐linked TAF1 mutations, which rarely can also manifest in women. Clinical and genetic heterogeneity also characterizes myoclonus‐dystonia, which includes not only the classical phenotype associated with epsilon‐sarcoglycan mutations but rarely also presentation of ANO3 gene mutations, TITF1 gene mutations typically underlying benign hereditary chorea, and some dopamine synthesis pathway conditions due to GCH1 and TH mutations. Thus, new genes are being recognized for isolated dystonia, and the phenotype of known genes is broadening and now involves different combined dystonia syndromes.
Expression of oxytocin receptors is greatly reduced in the placenta of heavy mares with retained fetal membranes due to secondary uterine atony
Authors: Rapacz‐Leonard A. , Raś A. , Całka J. , Janowski T. E.
Published: January 2015

Reasons for performing study

Fetal membrane retention can be a life‐threatening condition and its incidence exceeds 50% in heavy draught mares. Although fetal membrane retention is commonly treated with repeated injections of oxytocin, based on the suggestion that it is caused mainly by secondary atony of the uterus, this treatment sometimes fails. This led us to ask if expression of oxytocin receptors differs in mares that retain fetal membranes due to secondary uterine atony.

Objectives

To determine whether expression of oxytocin receptors in equine placental tissues differs when heavy draught mares expel fetal membranes or retain them because of secondary uterine atony.

Study design

Controlled study using archived tissues.

Methods

Placental biopsies (containing the endometrium and allantochorion) were taken from 8 heavy draught mares during parturition. Four mares expelled fetal membranes shortly after foaling (control mares) and 4 mares retained them (expulsion time was >3 h from delivery). The 4 mares that retained fetal membranes had secondary atony of the uterus. The amount of oxytocin receptors was estimated by measuring the intensity of western blot bands. The presence and location of oxytocin receptors were determined by immunocytochemistry.

Results

Oxytocin receptor expression was nearly 50 times less intense in mares with placenta retention due to secondary atony of the uterus and immunocytochemical staining was barely visible. In the control mares, oxytocin receptors were found in both epithelial and endothelial cells of the placenta and staining was most intense where the endometrium contacts the allantochorion.

Conclusions

Inadequate expression of oxytocin receptors may be a cause of uterine atony leading to fetal membrane retention.
Bile salt recognition by human liver fatty acid binding protein
Authors: Favretto Filippo , Santambrogio Carlo , D'Onofrio Mariapina , Molinari Henriette , Grandori Rita , Assfalg Michael
Published: January 2015

Fatty acid binding proteins (FABPs) act as intracellular carriers of lipid molecules, and play a role in global metabolism regulation. Liver FABP (L‐FABP) is prominent among FABPs for its wide ligand repertoire, which includes long‐chain fatty acids as well as bile acids (BAs). In this work, we performed a detailed molecular‐ and atomic‐level analysis of the interactions established by human L‐FABP with nine BAs to understand the binding specificity for this important class of cholesterol‐derived metabolites. Protein–ligand complex formation was monitored using heteronuclear NMR, steady‐state fluorescence spectroscopy, and mass spectrometry. BAs were found to interact with L‐FABP with dissociation constants in the narrow range of 0.6–7 μm; however, the diverse substitution patterns of the sterol nucleus and the presence of side‐chain conjugation resulted in complexes endowed with various degrees of conformational heterogeneity. Trihydroxylated BAs formed monomeric complexes in which single ligand molecules occupied similar internal binding sites, based on chemical‐shift perturbation data. Analysis of NMR line shapes upon progressive addition of taurocholate indicated that the binding mechanism departed from a simple binary association equilibrium, and instead involved intermediates along the binding path. The co‐linear chemical shift behavior observed for L‐FABP complexes with cholate derivatives added insight into conformational dynamics in the presence of ligands. The observed spectroscopic features of L‐FABP/BA complexes, discussed in relation to ligand chemistry, suggest possible molecular determinants of recognition, with implications regarding intracellular BA transport. Our findings suggest that human L‐FABP is a poorly selective, universal BA binder.
Mycobacterium tuberculosis RecA is indispensable for inhibition of the mitogen‐activated protein kinase‐dependent bactericidal activity of THP‐1‐derived macrophages in vitro
Authors: Szulc‐Kielbik Izabela , Brzezinska Marta , Kielbik Michal , Brzostek Anna , Dziadek Jaroslaw , Kania Katarzyna , Sulowska Zofia , Krupa Agnieszka , Klink Magdalena
Published: January 2015

Our knowledge about the mechanisms utilized by Mycobacterium tuberculosis to survive inside macrophages is still incomplete. One of the mechanism that protects M. tuberculosis from the host's microbicidal products and allows bacteria to survive involves DNA repair systems such as the homologous recombination (HR) and nonhomologous end‐joining (NHEJ) pathways. It is accepted that any pathway that contributes to genome maintenance should be considered as potentially important virulence factor. In these studies, we investigated reactive oxygen species, nitric oxide and tumor necrosis factor‐α production by macrophages infected with wild‐type M. tuberculosis, with an HR‐defective mutant (∆recA), with an NHEJ‐defective mutant [∆(ku,ligD)], with a mutant defective for both HR and NHEJ [∆(ku,ligD,recA)], or with appropriate complemented strains. We also assessed the involvement of extracellular signal‐regulated kinases (ERKs) 1 and 2 in the response of macrophages to infection with the above‐mentioned strains, and ERK1/2 phosphorylation in M. tuberculosis‐infected macrophages. We found that mutants lacking RecA induced a greater bactericidal response by macrophages than did the wild‐type strain or an NHEJ‐defective mutant, and activated ERK1/2 was involved only in the response of macrophages to recA deletion mutants [∆(ku,ligD,recA) and ∆recA]. We also demonstrated that only the triple mutant induced ERK1/2 phosphorylation in phorbol‐12‐myristate‐13‐acetate‐stimulated macrophages. Moreover, HR‐defective mutants induced lower amounts of tumor necrosis factor‐α secretion than did the wild‐type or ∆(ku,ligD). Our results indicate that RecA contributes to M. tuberculosis virulence, and also suggest that diminished ERK1/2 activation in macrophages infected with M. tuberculosis possessing recA may be an important mechanism by which wild‐type mycobacteria escape intracellular killing.
A Creative Helicobacter pylori Diagnosis Scheme Based on Multiple Genetic Analysis System: Qualification and Quantitation
Authors: Zhou Lifang , Zhao Fuju , Hu Binjie , Fang Yi , Miao Yingxin , Huang Yiqin , Ji Da'nian , Zhang Jinghao , Xu Lingli , Zhang Yanmei , Bao Zhijun , Zhao Hu
Published: January 2015

Background

Currently, several diagnostic assays for Helicobacter pylori (H. pylori) are available, but each has some limitations. Further, a high‐flux quantitative assay is required to assist clinical diagnosis and monitor the effectiveness of therapy and novel vaccine candidates.

Methods

Three hundred and eighty‐seven adult patients [nonulcer dyspepsia (NUD) 295, peptic ulcer disease (PUD) 77, gastric cancer (GC) 15] were enrolled for gastrointestinal endoscopies. Three biopsy samples from gastric antrum were collected for the following tests: culture, rapid urease test (RUT), histopathology, conventional polymerase chain reaction (PCR), and Multiple Genetic Analysis System (MGAS). The diagnostic capability of H. pylori for all methods was evaluated through the receiver operating characteristic (ROC) curves.

Results

Based on the gold standard, the sensitivity and specificity of MGAS were 92.9 and 92.4%, and positive predict value (PPV) and negative predict value (NPV) were 96.0 and 87.1%, respectively. All the above parameters of MGAS were higher than that of culture (except its specificity), RUT and histopathology, and nearly closed to that of conventional PCR. The area under curve (AUC) was 0.7575 (Culture), 0.8870 (RUT), 0.9000 (Histopathology), 0.9496 (Conventional PCR), and 0.9277 (MGAS). No significant statistical difference was observed for the H. pylori DNA load in different disease groups (p = .067). In contrast, a statistically significant difference in the H. pylori DNA copy number was observed based on age (p = .043) and gender (p = .021).

Conclusions

The data showed that MGAS performed well in detecting H. pylori infection. Furthermore, the quantitative analysis showed that the load of H. pylori was significantly different within both age and gender groups. These results suggested that MGAS could be a potential alternative method for clinical detection and monitoring of the effectiveness of H. pylori therapy.
Optimisation of soya bean oil bleaching by ultrasonic processing and investigate the physico‐chemical properties of bleached soya bean oil
Authors: Abedi Elahe , Sahari Mohammad Ali , Barzegar Mohsen , Azizi Mohammad Hossein
Published: January 2015

In this study, an ultrasonic bleaching method for soya bean oil under different conditions was optimised by monitoring the reduction of oil‐containing colourants. Ultrasonic power, clay content, temperature and time had a significant interaction effect on (P < 0.05) the reduction of oil pigments and colourants. Under optimised conditions, red and yellow colour indices in the ultrasound‐bleached soya bean oil reduced to 74.44% and 75.55%, and 54.18% and 58.85%, respectively, at amplitudes of 85% and 65%. Carotenoid and chlorophyll concentrations also reduced to 81.19% and 83.62%, and 94.66% and 95.26%, respectively, at 85% and 65% ultrasonic power. The overall contents of sterols and tocopherols decreased using ultrasonic bleaching process compared with the neutralised soya bean oil. Although the peroxide value (PV) of the soya bean oil after ultrasonic and control bleaching reduced, this index was higher in the ultrasonic bleached oil than the control one. The results showed that the ultrasonic bleaching process could be an alternative to the industrial bleaching method as it reduced clay usage, temperature and time of bleaching about 35%, 35% and 10%, respectively.
Isolation of antimicrobial agent from the marine algae Cystoseira hakodatensis
Authors: Jun Joon‐Young , Nakajima Sosuke , Yamazaki Koji , Kawai Yuji , Yasui Hajime , Konishi Yasuyuki
Published: January 2015

Cystoseira hakodatensis is an unutilised brown algae belonging to family Sargassaceae. A crude methanol extract from the algae showed inhibitory effects on the growths of Bacillus cereus and Bacillus licheniformis. To isolate the major antimicrobial agent, a sequential active‐guided isolation procedure was applied: liquid–liquid extraction, column chromatography and bio‐autography. A marked antimicrobial agent (active α) was isolated in hydrophobic fraction and was determined to phenolics without carbohydrates and proteins by phytochemical test. Regarding the antimicrobial potential, the isolated active α showed better inhibitory effects against B. cereus and B. licheniformis at 2 and 4 times of lower concentrations (62.5 and 31.3 μg mL−1) in comparison with epigallocatechin gallate. These results showed that C. hakodatensis is a potential source of antimicrobial agent capable of preventing the growth of the two bacteria.
Polymorphisms of the NOS3 gene in Tunisian patients with Behçet's disease
Authors: Kallel A. , Sbaï M. H. , Houman M. H. , Sediri Y. , Ouertani D. , Smiti Khanfir M. , Ben Ghorbel I. , Jemaa R. , Kaabachi N.
Published: January 2015

Behçet's disease (BD) is a multisystem inflammatory disease characterized by recurrent orogenital ulceration, ocular inflammation and skin lesions. Reduced plasma nitric oxide (NO) levels in patients with BD have been implicated in the development of the endothelial abnormalities and thrombotic complications occurring in these patients. Polymorphisms in the endothelial nitric oxide synthase gene (NOS3) have been inconsistently associated with BD. This inconsistency may derive from population stratification secondary to ethnic diversity, and consideration limited to only one rather than combinations of polymorphisms. We studied three genetic variations in the NOS3 gene: a single nucleotide polymorphism in the promoter region −786T>C, in exon 7 (Glu298Asp), and a variable number of tandem repeats in intron 4 (4a4b) of the NOS3 gene in 100 unrelated Tunisian patients with BD and 148 healthy controls. In addition, we also examined the association of NOS3 gene haplotypes with BD. Analyses of the Glu298Asp, −786T>C and 4a4b polymorphisms were made by the polymerase chain reaction (PCR) restriction fragment length polymorphism technique and PCR genotyping, respectively. The distribution of the Glu298Asp genotype differed significantly between patients with BD and controls (P = 0.01). Allele Asp298 was significantly more frequent in patients with BD than in controls (P = 0.005, OR = 1.70, 95% CI 1.14–2.54). In contrast, distribution of alleles and genotypes of −786T>C and 4a4b polymorphisms was not different between the control and BD group. However, the frequency of Asp‐T‐4b haplotype was significantly higher in patients with BD than in healthy controls. By gender, the signification remained only for heterozygous men (P = 0.03) and homozygous women (P = 0.02). These results suggest that Glu298Asp polymorphism of the NOS3 gene is associated with BD susceptibility in Tunisian patients.
Mesectoderm is a major target of retinoic acid action
Authors: Mark Manuel , Ghyselinck Norbert B. , Kastner Philippe , Dupé Valérie , Wendling Olivia , Krezel Wojciech , Mascrez Bénédicte , Chambon Pierre
Published: January 2015

The RAR and RXR families of retinoid nuclear receptors each comprise three isotypes (α, β and γ). In vitro, RARs bind Co their cognate DNA response elements as heterodimers with RXRs. Null mutations of all six isotypes have been generated. The defects displayed by RARα, β and γ single null mutant mice are confined to a small subset of the tissues normally expressing these receptors. This discrepancy reflects the existence of a functional redundancy, since RAR double null mutants exhibit congenital malformations in almost every organ system. In particular, most of the structures derived from the mesectoderm are severely affected. Analysis of mutant mice lacking both RARs and RXRs indicates that RXRα: RARγ heterodimers are instrumental in the patterning of craniofacial skeletal elements, whereas RXRα: RARα heterodimers may be preferentially involved in the generation of neural crest cell‐derived arterial smooth muscle cells. Both RXRα: RARβ and RXRα: RARγ heterodimers appear to function during the development of the ocular mesenchyme. Moreover, atavistic reptilian cranial structures are generated in RAR mutants, suggesting that the RA signal has been implicated in the modification of developmental programs in the mesectoderm during evolution.
Molecular control of odontogenic patterning: positional dependent initiation and morphogenesis
Authors: Thomas Bethan L. , Tucker Abigail S. , Ferguson Christine , Qiu Mensheng , Rubenstein John L. R. , Sharpe Paul T.
Published: January 2015

We have previously proposed that the patterning of the mammalian dentition is determined by an odontogenic homeobox code, whereby positional specification of odontogenic ectomesenchymal cell populations in a given region of the first branchial arch are determined by the combination of different homeobox genes expressed, described in detail in (1). Here we describe the first functional evidence for such a mechanism, and show that development of different types of teeth is controlled by independent genetic pathways. We suggest a mechanism whereby patterning is determined by position dependent control of tooth germ initiation, and propose that the pathway of morphogenesis is directly linked to the early mesenchymal expression of homeobox genes.
Patterning of the murine dentition by homeobox genes
Authors: Thomas Bethan L. , Sharpe Paul T.
Published: January 2015

Homeobox genes have been shown to be important for the regulation of pattern formation of many systems during embryogenesis. Overlapping domains of Hox gene expression in the paraxial mesoderm have been suggested to create a combinatorial code of expression (Hox code) specifying the structures of individual segments such as the vertebrae. Hox genes are not expressed in the neural crest cells contributing to tooth formation, and so a Hox code can not be involved in patterning the dentition. It has previously been proposed that other, non‐Hox homeobox genes may pattern the dentition. Expression data in this paper shows that there is a pattern of overlapping domains of homeobox gene expression in facial mesenchyme prior to the initiation of tooth development. We propose that expression of these genes constitutes an odontogenic homeobox code which patterns the dentition.
Immunolocalisation of collagens in the developing rat molar tooth
Authors: Webb Philip Patrick , Moxham Bernard John , Ralphs James Robert , Benjamin Michael
Published: January 2015

This study identifies different types of collagens during tooth development, maturation and ageing. Tissues from the rat first molar (from animals ranging in age from E14 to 104 wk postnatally) were immunostained using a panel of mono‐ and polyclonal antibodies against types I, II, III, IV, VI, IX and X collagen, fibronectin and laminin. During tooth development, types I and III collagens were expressed in the dental papilla at all stages but were also unexpectedly observed in the stellate reticulum of the enamel organ. Transient expression of type II collagen was also observed in the stellate reticulum during the late bell stage. Types IV and VI collagens, with laminin and fibronectin, were located within the basement membranes of the tooth germ. Collagen types I and III were observed within the developing follicle/periodontal ligament, type III predominating where collagen fibres were inserting into the alveolar bone and cementum. The pattern of types I and III collagen labelling within the periodontal ligament and the dental pulp did not change with age. Thus, some unusual collagen localisations were observed in the tooth germ, particularly within the stellate reticulum.
The bone morphogenetic protein family: multifunctional cellular regulators in the embryo and adult
Authors: Wozney John M.
Published: January 2015

Originally identified as the active components within osteoinductive extracts derived from bone, the BMPs now are known to include a large family of proteins within the TGF‐β superfamily of growth and differentiation factors. Members of the BMP family have been determined to be key signaling molecules in embryogenesis, in species ranging from Drosophila to humans. They are involved in delivering positional information, the development of hard tissues (both bones and teeth), as well as soft tissue types. When implanted into adult animals, several of the BMPs have been shown to initiate the complex cellular process resulting in the induction of bone through both the endochondral and intramembranous bone formation pathways. Preclinical studies have shown the ability of these factors to induce bone and heal large bony defects in a variety of models relevant to clinical problems in orthopedics, as well as the oral and maxillofacial dental areas. For example, implantation of recombinant human BMP‐2 (rhBMP‐2) has been shown to augment the alveolar ridge in several animal models, and when placed in a periodontal environment to restore not only new bone, but also the attachment tissues. Results from ongoing clinical studies support the ability of rhBMP‐2 implants to induce physiologic bone.
Activin and bone morphogenetic protein (BMP) signalling during tooth development
Authors: Heikinheimo Kristiina , Bègue‐Kirn Catherine , Ritvos Olli , Tuuri Timo , Ruch Jean Victor
Published: January 2015

Activins and bone morphogenetic proteins (BMPs) belong to the transforming growth factor‐β (TGF‐β) superfamily of structurally related cytokines. Results of recent in vivo and in vitro studies suggest that activin A, several BMPs, and their cell surface receptors may participate in tooth development. In addition, follistatin, an extracellular protein that may interact with activin and BMP signaling, appears to be involved. This review focuses on recent advances in relation to the roles of activin/BMP receptors, their ligands and extracellular modifiers during tooth development.
Role of exogenous TGF‐β in induction of reparative dentinogenesis in vivo
Authors: Tziafas Dimitrios , Papadimitriou Serafim
Published: January 2015

This study was designed to investigate in vivo i) the role of TGF‐β as an active component of the dentin matrix during induction of reparative dentinogenesis; and ii) the ability of TGF‐β1 isoform to induce dentinogenic events. Dental pulps of dog molars and canines were mechanically exposed, and the following implants were placed intrapulpally for 42 d: i) Demineralized or native autogenous dentin matrix preincubated with a TGF‐β‐neutralizing antibody; ii) Millipore filters soaked with solution containing 100 ng/ml of TGF‐β1 from human platelets; iii) biomatrices and filters soaked in control solutions. After incubation of biomatrices with the TGF‐β‐neutralizing antibody, demineralized dentin completely lost its inductive activity, while native dentin was only able to stimulate formation of fibrodentin. Millipore filters soaked with TGF‐β1 were consistently surrounded by a thick zone of tubular matrix lined with high columnar polarized cells. The data provide evidence that pulp cells can express their dentinogenic potential in response to an appropriate surface containing exogenous TGF‐β1, and that the dentinogenic activity of dentin matrix may at least partly be ascribed to TGF‐β molecule(s).