bluuebunny/crossref_metadata_2025 huggingface dataset:
Familial Atypical Hemolytic Uremic Syndrome: A Review of Its Genetic and Clinical Aspects
Authors: Bu Fengxiao , Borsa Nicolo , Gianluigi Ardissino , Smith Richard J. H.
Published: November 2012

Atypical hemolytic uremic syndrome (aHUS) is a rare renal disease (two per one million in the USA) characterized by microangiopathic hemolytic anemia, thrombocytopenia, and acute renal failure. Both sporadic (80% of cases) and familial (20% of cases) forms are recognized. The study of familial aHUS has implicated genetic variation in multiple genes in the complement system in disease pathogenesis, helping to define the mechanism whereby complement dysregulation at the cell surface level leads to both sporadic and familial disease. This understanding has culminated in the use of Eculizumab as first-line therapy in disease treatment, significantly changing the care and prognosis of affected patients. However, even with this bright outlook, major challenges remain to understand the complexity of aHUS at the genetic level. It is possible that a more detailed picture of aHUS can be translated to an improved understanding of disease penetrance, which is highly variable, and response to therapy, both in the short and long terms.
Seismic Hazard Assessment for the Tianshui Urban Area, Gansu Province, China
Authors: Wang Zhenming , Butler David T. , Woolery Edward W. , Wang Lanmin
Published: November 2012

A scenario seismic hazard analysis was performed for the city of Tianshui. The scenario hazard analysis utilized the best available geologic and seismological information as well as composite source model (i.e., ground motion simulation) to derive ground motion hazards in terms of acceleration time histories, peak values (e.g., peak ground acceleration and peak ground velocity), and response spectra. This study confirms that Tianshui is facing significant seismic hazard, and certain mitigation measures, such as better seismic design for buildings and other structures, should be developed and implemented. This study shows that PGA of 0.3 g (equivalent to Chinese intensity VIII) should be considered for seismic design of general building and PGA of 0.4 g (equivalent to Chinese intensity IX) for seismic design of critical facility in Tianshui.
Focus on Therapeutic Strategies of Nonalcoholic Fatty Liver Disease
Authors: Durazzo Marilena , Belci Paola , Collo Alessandro , Grisoglio Enrica , Bo Simona
Published: November 2012

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease in the Western world (it affects 30% of the general adult population). The NAFLD encompasses a histological spectrum ranging from simple steatosis to nonalcoholic steatohepatitis (NASH), defined by steatosis, hepatocellular damage, and lobular inflammation in individuals without significant alcohol consumption and negative viral, congenital, and autoimmune liver disease markers. Currently, NAFLD is considered an emerging epidemic in light of the dramatic increase in obesity rates. With the progressive nature of NASH and its rising prevalence there is a significant need for a specific and targeted treatments since to date there has not been any validated therapies for NAFLD other than weight loss, which is well known to have a poor long-term success rate. In recent years, visceral adipose tissue has taken an important role in NAFLD pathogenesis, and current therapeutic approaches aim at reducing visceral obesity and free fatty acid overflow to the liver. This paper is focused on the treatments used for NAFLD and the potential new therapy.
Antioxidant Activity and Hepatoprotective Potential ofPolyalthia longifoliaandCassia spectabilisLeaves against Paracetamol-Induced Liver Injury
Authors: Jothy Subramanion L. , Aziz Azlan , Chen Yeng , Sasidharan Sreenivasan
Published: November 2012

In the present study,in vitroantioxidant, free radical scavenging capacity, and hepatoprotective activity of methanol extracts fromPolyalthia longifoliaandCassia spectabiliswere evaluated using establishedin vitromodels such as ferric-reducing antioxidant power (FRAP), 2,2-diphenyl-1-picryl-hydrazyl(DPPH•), hydroxyl radical(OH•), nitric oxide radical(NO•)scavenging, metal chelating, and antilipidperoxidation activities. Interestingly, all the extracts showed considerablein vitroantioxidant and free radical scavenging activities in a dose-dependent manner when compared to the standard antioxidant which verified the presence of strong antioxidant compound in leaf extracts tested. Phenolic and flavonoid content of these extracts is significantly correlated with antioxidant capacity. SinceP. longifoliaextract was exhibited betterin vitroantioxidant activities, it was subjected forin vivohepatoprotective activity in paracetamol-intoxicated mice. Therapy ofP. longifoliashowed the liver protective effect on biochemical and histopathological alterations. Moreover, histological studies also supported the biochemical finding, that is, the maximum improvement in the histoarchitecture of the liver. Results revealed thatP. longifolialeaf extract could protect the liver against paracetamol-induced oxidative damage by possibly increasing the antioxidant protection mechanism in mice. Our findings indicated thatP. longifoliaandC. spectabilishave potential as good sources of natural antioxidant/antiaging compounds.
Inhibition of Connexin 26/43 and Extracellular-Regulated Kinase Protein Plays a Critical Role in Melatonin Facilitated Gap Junctional Intercellular Communication in Hydrogen Peroxide-Treated HaCaT Keratinocyte Cells
Authors: Lee Hyo-Jung , Lee Hyo-Jeong , Sohn Eun Jung , Lee Eun-Ok , Kim Jin-Hyoung , Lee Min-Ho , Kim Sung-Hoon
Published: November 2012

Though melatonin was known to regulate gap junctional intercellular communication (GJIC) in chick astrocytes and mouse hepatocytes, the underlying mechanism by melatonin was not elucidated in hydrogen peroxide- (H2O2-) treated HaCaT keratinocyte cells until now. In the current study, though melatonin at 2 mM and hydrogen peroxide (H2O2) at 300 μM showed weak cytotoxicity in HaCaT keratinocyte cells, melatonin significantly suppressed the formation of reactive oxygen species (ROS) in H2O2-treated HaCaT cells compared to untreated controls. Also, the scrape-loading dye-transfer assay revealed that melatonin enhances the intercellular communication by introducing Lucifer Yellow into H2O2-treated cells. Furthermore, melatonin significantly enhanced the expression of connexin 26 (Cx26) and connexin 43 (Cx43) at mRNA and protein levels, but not that of connexin 30 (Cx30) in H2O2-treated HaCaT cells. Of note, melatonin attenuated the phosphorylation of extracellular signal-regulated protein kinases (ERKs) more than p38 MAPK or JNK in H2O2-treated HaCaT cells. Conversely, ERK inhibitor PD98059 promoted the intercellular communication in H2O2-treated HaCaT cells. Furthermore, combined treatment of melatonin (200 μM) and vitamin C (10 μg/mL) significantly reduced ROS production in H2O2-treated HaCaT cells. Overall, these findings support the scientific evidences that melatonin facilitates gap junctional intercellular communication in H2O2-treated HaCaT keratinocyte cells via inhibition of connexin 26/43 and ERK as a potent chemopreventive agent.
Traditional Herbal Management of Sickle Cell Anemia: Lessons from Nigeria
Authors: Ameh Sunday J. , Tarfa Florence D. , Ebeshi Benjamin U.
Published: November 2012

Background. Patients in West Africa where sickle cell anemia (SCA) is endemic have for ages been treated with natural products, especially herbs, as, is still the case in rural communities.Objective. In this paper we look closely at some of these herbs to see if there are any lessons to be learnt or clues to be found for optimizing the treatments based on them, as had been done in the case of NIPRISAN, which was developed from herbs in Nigeria based on Yoruba Medicine.Methods. Select publications on SCA, its molecular biology and pathology, and actual and experimental cases of herbal treatment were perused in search of molecular clues that can be linked to chemical constituents of the herbs involved.Results. The study revealed that during the last 2-3 decades, much progress was made in several aspects of SCA pharmacology, especially the approval of hydroxyurea. As for SCA herbalism, this paper revealed that antisickling herbs abound in West Africa and that the most promising may yet be found. Three new antisickling herbs (Entandrophragma utile,Chenopodium ambrosioides, andPetiveria alliacea) were reported in May 2011. At NIPRD, where NIPRISAN was developed, three other recipes are currently awaiting development.Conclusion. The study raised the hope that the search in the Tropics for more effective herbal recipes for managing sickle cell anaemia will be more fruitful with time and effort.
Inhibitory Activities ofZygophyllum album: A Natural Weight-Lowering Plant on Key Enzymes in High-Fat Diet-Fed Rats
Authors: Mnafgui Kais , Hamden Khaled , Ben Salah Hichem , Kchaou Mouna , Nasri Mbarek , Slama Sadok , Derbali Fatma , Allouche Noureddine , Elfeki Abdelfattah
Published: November 2012

Obesity is a serious health problem that increased risk for many complications, including diabetes and cardiovascular disease. The results showed EZA, which found rich in flavonoids and phenolic compounds, exhibited an inhibitory activity on pancreatic lipasein vitrowith IC50of 91.07 μg/mL.In vivoadministration of this extract to HFD-rats lowered body weight and serum leptin level; and inhibited lipase activity of obese rats by 37% leading to notable decrease of T-Ch, TGs and LDL-c levels accompanied with an increase in HDL-c concentration in serum and liver of EZA treated HFD-rats. Moreover, the findings revealed that EZA helped to protect liver tissue from the appearance of fatty cysts. Interestingly, supplementation of EZA modulated key enzyme related to hypertension such as ACE by 36% in serum of HFD animals and improve some of serum electrolytes such as Na+, K+, Cl−, Ca2+and Mg2+. Moreover, EZA significantly protected the liver-kidney function by reverted back near to normal the values of the liver-kidney dysfunction indices AST&ALT, ALP, CPK and GGT activities, decreased T-Bili, creat, urea and uric acid rates. In conclusion, these results showed a strong antihypelipidemic effect of EZA which can delay the occurrence of dislipidemia and hypertension.
Nanobiotechnological Nanocapsules Containing Polyhemoglobin-Tyrosinase: Effects on Murine B16F10 Melanoma Cell Proliferation and Attachment
Authors: Wang Yun , Chang Thomas M. S.
Published: November 2012

We have reported previously that daily intravenous infusions of a soluble nanobiotechnological complex, polyhemoglobin-tyrosinase [polyHb-Tyr], can suppress the growth of murine B16F10 melanoma in a mouse model. In order to avoid the need for daily intravenous injections, we have now extended this further as follows. We have prepared two types of biodegradable nanocapsules containing [polyHb-Tyr]. One type is to increase the circulation time and decrease the frequency of injection and is based on polyethyleneglycol-polylactic acid (PEG-PLA) nanocapsules containing [polyHb-Tyr]. The other type is to allow for intratumoural or local injection and is based on polylactic acid (PLA) nanocapsules containing [polyHb-Tyr]. Cell culture studies show that it can inhibit the proliferation of murine B16F10 melanoma cells in the “proliferation model”. It can also inhibit the attachment of murine B16F10 melanoma cells in the “attachment model.” This could be due to the action of tyrosinase on the depletion of tyrosine or the toxic effect of tyrosine metabolites. The other component, polyhemoglobin (polyHb), plays a smaller role in nanocapsules containing [polyHb-Tyr], and this is most likely by its depletion of nitric oxide needed for melanoma cell growth.
A Case Study of Probit Model Analysis of Factors Affecting Consumption of Packed and Unpacked Milk in Turkey
Authors: Uzunoz Meral , Akcay Yasar
Published: November 2012

This paper focused on the effects of some sociodemographic factors on the decision of the consumer to purchase packed or unpacked fluid milk in Sivas, Turkey. The data were collected from 300 consumers by using face-to-face survey technique. The sample size was determined using the possibility-sampling method. Probit model has been used to analyze the socioeconomic factors affecting milk consumption of households. Four estimators (household size, income, milk preferences reason, and milk price) in the probit model were found statistically significant. According to empirical results, consumers with lower household size and higher income levels tend to consume packed milk consumption. Our study findings suggest that consumers who were sensitive to price were less likely to consume packed milk and believe that packed milk price is expensive compared to unpacked milk price. Also, milk price was effective factor concerning packed and unpacked milk consumption behavior. The majority of consumers read the contents of packed fluid milk and are affected by safety food in their shopping preferences.
Characterization of Blood Oxidative Stress in Type 2 Diabetes Mellitus Patients: Increase in Lipid Peroxidation and SOD Activity
Authors: Bandeira Suziy de M. , Guedes Glaucevane da S. , Fonseca Lucas José S. da , Pires André S. , Gelain Daniel P. , Moreira José Claudio F. , Rabelo Luíza A. , Vasconcelos Sandra Mary L. , Goulart Marília Oliveira F.
Published: November 2012

This study evaluated the oxidative stress through enzymatic and nonenzymatic biomarkers in diabetic patients with and without hypertension and prediabetics. The SOD and CAT (in erythrocytes) and GPx (in plasma) enzymatic activities, plasma levels of lipid peroxidation, and total thiols were measured in the blood of 55 subjects with type 2 diabetes and 38 subjects without diabetes (9 pre-diabetics and 29 controls) aged 40–86 years. The total SOD activity and the lipid peroxidation were higher in diabetics compared to nondiabetics. In stratified groups, the total SOD activity was different for the hypertensive diabetics compared to the prediabetics and normotensive controls. Lipid peroxidation was significantly higher in both groups of diabetics (hypertensive and normotensive) compared to prediabetic groups and hypertensive and normotensive controls. There was no significant difference in the CAT and GPx activities, as well as in the concentration of total thiols in the groups studied. Present data strongly suggest the involvement of oxidative stress in the pathophysiology of diabetes, revealing that the increased lipid peroxidation has a close relationship with high glucose levels, as observed by the fasting glucose and HbA1c levels. The results evidence the correlation between lipid peroxidation and DM, irrespective of the presence of hypertension.
Closing the Window on Light Charged Higgs Bosons in the NMSSM
Authors: Rathsman Johan , Rössler Thomas
Published: November 2012

In the Next-to-Minimal SuperSymmetric Model (NMSSM) the lightest CP-odd Higgs bosons (a1) can be very light. As a consequence, in addition to the standard charged Higgs boson (h±) decays considered in the MSSM for a light charged Higgs (mh±<mt), the branching fraction forh±→a1Wcan be dominant. We investigate how this signal can be searched for intt-production at the Large Hadron Collider (LHC) in the case that (ma1≳2mB) with thea1giving rise to a singlebb--jet and discuss to what extent the LHC experiments are able to discover such a scenario with an integrated luminosity ~20 fb−1. We also discuss the implications of the possible Higgs-signal observed at the LHC.
Phase-Conjugate-State Pairs in Entangled States
Authors: Namiki Ryo
Published: November 2012

We consider the probability that a bipartite quantum state contains phase-conjugate-state (PCS) pairs and/or identical-state pairs as signatures of quantum entanglement. While the fraction of the PCS pairs directly indicates the property of a maximally entangled state, the fraction of the identical-state pairs negatively determines antisymmetric entangled states such as singlet states. We also consider the physical limits of these probabilities. This imposes fundamental restrictions on the pair appearance of the states with respect to the local access of the physical system. For continuous-variable system, we investigate similar relations by employing the pairs of phase-conjugate coherent states. We also address the role of the PCS pairs for quantum teleportation in both discrete-variable and continuous-variable systems.
Importance of the Brain Angiotensin System in Parkinson’s Disease
Authors: Wright John W. , Harding Joseph W.
Published: November 2012

Parkinson’s disease (PD) has become a major health problem affecting 1.5% of the world’s population over 65 years of age. As life expectancy has increased so has the occurrence of PD. The primary direct consequence of this disease is the loss of dopaminergic (DA) neurons in the substantia nigra and striatum. As the intensity of motor dysfunction increases, the symptomatic triad of bradykinesia, tremors-at-rest, and rigidity occur. Progressive neurodegeneration may also impact non-DA neurotransmitter systems including cholinergic, noradrenergic, and serotonergic, often leading to the development of depression, sleep disturbances, dementia, and autonomic nervous system failure. L-DOPA is the most efficacious oral delivery treatment for controlling motor symptoms; however, this approach is ineffective regarding nonmotor symptoms. New treatment strategies are needed designed to provide neuroprotection and encourage neurogenesis and synaptogenesis to slow or reverse this disease process. The hepatocyte growth factor (HGF)/c-Met receptor system is a member of the growth factor family and has been shown to protect against degeneration of DA neurons in animal models. Recently, small angiotensin-based blood-brain barrier penetrant mimetics have been developed that activate this HGF/c-Met system. These compounds may offer a new and novel approach to the treatment of Parkinson’s disease.
Modelling of the Curvature Term of the Flame Surface Density Transport Equation for Large Eddy Simulations
Authors: Katragadda Mohit , Chakraborty Nilanjan
Published: November 2012

A simplified chemistry based three-dimensional Direct Numerical Simulation (DNS) database of freely propagating statistically planar turbulent premixed flames with a range of different values of turbulent Reynolds number has been used for the a priori modelling of the curvature term of the generalised Flame Surface Density (FSD) transport equation in the context of Large Eddy Simulation (LES). The curvature term has been split into the contributions arising due to the reaction and normal diffusion components of displacement speed and the term originating from the tangential diffusion component of displacement speed. Subsequently, these contributions of the curvature term have been split into the resolved and subgrid contributions. New models have been proposed for the subgrid curvature terms arising from the combined reaction and normal diffusion components and the tangential diffusion component of displacement speed. The performances of the new model and the existing models for the subgrid curvature term have been compared with the corresponding quantity extracted from the explicitly filtered DNS data. The new model for the subgrid curvature term is shown to perform satisfactorily in all cases considered in the current study, accounting for wide variations in LES filter size.
Coupled TLC and MALDI-TOF/MS Analyses of the Lipid Extract of the Hyperthermophilic ArchaeonPyrococcus furiosus
Authors: Lobasso Simona , Lopalco Patrizia , Angelini Roberto , Vitale Rita , Huber Harald , Müller Volker , Corcelli Angela
Published: November 2012

The lipidome of the marine hyperthermophilic archaeonPyrococcus furiosuswas studied by means of combined thin-layer chromatography and MALDI-TOF/MS analyses of the total lipid extract. 80–90% of the major polar lipids were represented by archaeol lipids (diethers) and the remaining part by caldarchaeol lipids (tetraethers). The direct analysis of lipids on chromatography plate showed the presence of the diphytanylglycerol analogues of phosphatidylinositol and phosphatidylglycerol, theN-acetylglucosamine-diphytanylglycerol phosphate plus some caldarchaeol lipids different from those previously described. In addition, evidence for the presence of the dimeric ether lipid cardiolipin is reported, suggesting that cardiolipins are ubiquitous in archaea.
A Truncated Waveguide Fed by a Microstrip as a Radiating Element for High-Performance Automotive Anticollision Radars
Authors: Casula Giovanni Andrea , Mazzarella Giuseppe , Montisci Giorgio
Published: November 2012

A small truncated waveguide fed by a microstrip line through a transverse coupling slot is proposed and assessed as a high-performance antenna and array element in the K band and above. This antenna allows to obtain a high radiated power, with a very low cross-polar component in the radiated field. It is therefore particularly suitable for application in automotive anticollision radars. The proposed radiating element has been analyzed by a numerical code based on an in-house method of moments, and the microstrip feeding line has been modeled by its equivalent magnetic-wall waveguide. A linear array of such elements has been designed and matched with a BPF-inspired matching network allowing an in-band behavior suitable for anti-collision radar use, with an out-of-band rejection large enough to avoid the first receiving BPF.
Activated Carbon‐Fly Ash‐Nanometal Oxide Composite Materials: Preparation, Characterization, and Tributyltin Removal Efficiency
Authors: Ayanda Olushola S. , Fatoki Olalekan S. , Adekola Folahan A. , Ximba Bhekumusa J.
Published: November 2012

The physicochemical properties, nature, and morphology of composite materials involving activated carbon, fly ash, nFe3O4, nSiO2, and nZnO were investigated and compared. Nature and morphology characterizations were carried out by means of scanning electron and transmission electron microscopy, X‐ray diffraction, and Fourier transform infrared spectroscopy. Other physicochemical characterizations undertaken were CNH analysis, ash content, pH, point of zero charge, and surface area and porosity determination by BET. Experimental results obtained revealed that activated carbon, nSiO2, activated carbon‐fly ash, activated carbon‐fly ash‐nFe3O4, activated carbon‐fly ash‐nSiO2, and activated carbon‐fly ash‐nZnO composite materials exhibited net negative charge on their surfaces while fly ash, nFe3O4, and nZnO possessed net positive charge on their surfaces. Relatively higher removal efficiency (>99%) of TBT was obtained for all the composite materials compared to their respective precursors except for activated carbon. These composite materials therefore offer great potential for the remediation of TBT in wastewaters.
Synthesis and Antimicrobial Activity of Some [1,2,4]‐Triazole Derivatives
Authors: Patil Bhimagouda S. , Krishnamurthy G. , Shashikumar N. D. , Lokesh M. R. , Bhojya Naik H. S.
Published: November 2012

A series of novel [1,2,4]‐triazolo piperidine (8), [1,2,4]‐triazolo piperazine (9a-c), [1,2,4]‐triazolo phenylether (10a-e), and [1,2,4]‐triazolo aniline (11a-c) derivatives have been synthesized. The chemical structures of the newly synthesized compounds were characterized by IR, 1H NMR, 13C NMR, and LCMS. The newly synthesized compounds were screened for antimicrobial activity. Among all the compounds tested, 11b (R4 = 4‐MeO–) showed the highest activity against Staphylococcus aureus and Escherichia coli, and 9a (R1 and R2 = Cl) showed the highest activity against Pseudomonas Aeruginosa.
Linear and Nonlinear Regression Methods for Equilibrium Modelling of p‐Nitrophenol Biosorption by Rhizopus oryzae: Comparison of Error Analysis Criteria
Authors: Yaneva Zvezdelina Lyubenova , Koumanova Bogdana Koumanova , Georgieva Nedyalka Valkanova
Published: November 2012

The study assessed the applicability of Rhizopus oryzae dead fungi as a biosorbent medium for p‐nitrophenol (p‐NP) removal from aqueous phase. The extent of biosorption was measured through five equilibrium sorption isotherms represented by the Langmuir, Freundlich, Redlich‐Peterson, multilayer and Fritz‐Schlunder models. Linear and nonlinear regression methods were compared to determine the best‐fitting equilibrium model to the experimental data. A detailed error analysis was undertaken to investigate the effect of applying seven error criteria for the determination of the single‐component isotherm parameters. According to the comparison of the error functions and to the estimation of the corrected Akaike information criterion (AICC), the Freundlich equation was ranked as the first and the Fritz‐Schlunder as the second best‐fitting models describing the experimental data. The present investigations proved the high efficiency (94%) of Rhizopus Oryzae as an alternative adsorbent for p‐NP removal from aqueous phase and revealed the mechanism of the separation process.
Antioxidant Activity and Isolation of Luteoline from Centaurea behen L. Grown in Iran
Authors: Esmaeili Akbar , Mousavi Zahra , Shokrollahi Maryam , Shafaghat Ali
Published: November 2012

Flavonoids are secondary metabolites providing Ultraviolet‐visible (UV) spectroscopy protection and color in almost all terrestrial plants and fruits. They have a fused ring system consisting of an aromatic ring and a benzopyran ring with a phenyl substituent. As their biological activities have an impact on human health, they serve as target molecules in the development of new drugs. The objective of this research was to study the antioxidant activity and chemical analysis of the luteoline from Centaurea behen L. (Compositae family). The aerial parts of powdered and dried C. behen were extracted with methanol (MeOH) in a Soxhlet apparatus over a period of 2 days. The concentrated total extract was extracted with petroleum ether, diethylether, and methanol. From the methanol extract of the aerial parts of C. behen, the flavonoid derivative (luteoline) was identified. The aerial parts’ extract demonstrated effective antioxidant activity measured in terms of half‐maximal inhibitory concentration (IC50). The product extract has been isolated by UV, column chromatography (CC), and preparative high‐performance liquid chromatography (HPLC). The structures involved were elucidated by 1H and 13C nuclear magnetic resonance (NMR) and heteronuclear multiple‐bond correlation (HMBC) spectra. The compound identified had not been reported in previous studies of C. behen L.
Genomic Profiling of Isolated Circulating Tumor Cells from Metastatic Breast Cancer Patients
Authors: Magbanua Mark Jesus M. , Sosa Eduardo V. , Roy Ritu , Eisenbud Lauren E. , Scott Janet H. , Olshen Adam , Pinkel Dan , Rugo Hope S. , Park John W.
Published: November 2012

Molecular characterization of circulating tumor cells (CTC) from blood is technically challenging because cells are rare and difficult to isolate. We developed a novel approach to isolate CTCs from blood via immunomagnetic enrichment followed by fluorescence-activated cell sorting (IE–FACS). Isolated CTCs were subjected to genome-wide copy number analysis via array comparative genomic hybridization (aCGH). In clinical studies, CTCs were isolated from 181 patients with metastatic breast cancer, 102 of which were successfully profiled, including matched archival primary tumor from five patients. CTCs revealed a wide range of copy number alterations including those previously reported in breast cancer. Comparison with two published aCGH datasets of primary breast tumors revealed similar frequencies of recurrent genomic copy number aberrations. In addition, serial testing of CTCs confirmed reproducibility and indicated genomic change over time. Comparison of CTCs with matched archival primary tumors confirmed shared lineage as well as some divergence. We showed that it is feasible to isolate CTCs away from hematopoietic cells with high purity through IE–FACS and profile them via aCGH analysis. Our approach may be used to explore genomic events involved in cancer progression and to monitor therapeutic efficacy of targeted therapies in clinical trials in a relatively noninvasive manner. Cancer Res; 73(1); 30–40. ©2012 AACR.
Overexpression of the Mitochondrial Folate and Glycine–Serine Pathway: A New Determinant of Methotrexate Selectivity in Tumors
Authors: Vazquez Alexei , Tedeschi Philip M. , Bertino Joseph R.
Published: November 2012

Previous studies have documented the roles of transport via the reduced folate carrier, retention via polyglutamylation, and increased levels of the target enzyme, dihydrofolate reductase in sensitivity to methotrexate. Recent studies have shown that the mitochondrial enzymes in the cellular metabolism of serine, folate, and glycine are overexpressed in a subset of human cancers and that their expression is required for tumor maintenance. In this Perspective article, we propose that the expression of mitochondrial enzymes in the metabolism of serine and glycine, in addition to those involved in folate metabolism, are determinants of the response to methotrexate. Furthermore, we show that myc activation in tumors is associated with upregulation of these enzymes. We propose that patients whose tumors show this phenotype will be sensitive to folate antagonists targeting thymidylate or purine biosynthesis. Cancer Res; 73(2); 478–82. ©2012 AACR.
Identification of Serum Proton NMR Metabolomic Fingerprints Associated with Hepatocellular Carcinoma in Patients with Alcoholic Cirrhosis
Authors: Nahon Pierre , Amathieu Roland , Triba Mohamed N. , Bouchemal Nadia , Nault Jean-Charles , Ziol Marianne , Seror Olivier , Dhonneur Gilles , Trinchet Jean-Claude , Beaugrand Michel , Le Moyec Laurence
Published: November 2012

Purpose: Metabolomics depicts metabolic changes in biologic systems using a multiparametric analysis technique. This study assessed the metabolomic profiles of serum, obtained by proton nuclear magnetic resonance (NMR) spectroscopy, from cirrhotic patients with and without hepatocellular carcinoma (HCC).

Experimental Design: The study included 154 consecutive patients with compensated biopsy-proven alcoholic cirrhosis. Among these, 93 had cirrhosis without HCC, 28 had biopsy-proven HCC within the Milan criteria and were eligible for curative treatment (small HCC), and 33 had HCC outside the Milan criteria (large HCC). Proton spectra were acquired at 500 MHz. An orthogonal partial latent structure [orthogonal projection to latent structure (OPLS)] analysis model was built to discriminate large HCC spectra from cirrhotic spectra. Small HCC spectra were secondarily projected using previously built OPLS discriminant components.

Results: The OPLS model showed discrimination between cirrhotic and large HCC spectra. Metabolites that significantly increased with large HCC were glutamate, acetate, and N-acetyl glycoproteins, whereas metabolites that correlated with cirrhosis were lipids and glutamine. Projection of small HCC samples into the OPLS model showed a heterogeneous distribution between large HCC and cirrhotic samples. Small HCC patients with metabolomic profile similar to those of large HCC group had higher incidences of recurrence or death during follow-up.

Conclusions: Serum NMR-based metabolomics identified metabolic fingerprints that could be specific to large HCC in cirrhotic livers. From a metabolomic standpoint, some patients with small HCC, who are eligible for curative treatments, seem to behave as patients with advanced cancerous disease. It would be useful to further prospectively investigate these patients to define a subgroup with a worse prognosis. Clin Cancer Res; 18(24); 6714–22. ©2012 AACR.
Expression Profiling of Archival Tumors for Long-term Health Studies
Authors: Waldron Levi , Ogino Shuji , Hoshida Yujin , Shima Kaori , McCart Reed Amy E. , Simpson Peter T. , Baba Yoshifumi , Nosho Katsuhiko , Segata Nicola , Vargas Ana Cristina , Cummings Margaret C. , Lakhani Sunil R. , Kirkner Gregory J. , Giovannucci Edward , Quackenbush John , Golub Todd R. , Fuchs Charles S. , Parmigiani Giovanni , Huttenhower Curtis
Published: November 2012

Purpose: More than 20 million archival tissue samples are stored annually in the United States as formalin-fixed, paraffin-embedded (FFPE) blocks, but RNA degradation during fixation and storage has prevented their use for transcriptional profiling. New and highly sensitive assays for whole-transcriptome microarray analysis of FFPE tissues are now available, but resulting data include noise and variability for which previous expression array methods are inadequate.

Experimental Design: We present the two largest whole-genome expression studies from FFPE tissues to date, comprising 1,003 colorectal cancer (CRC) and 168 breast cancer samples, combined with a meta-analysis of 14 new and published FFPE microarray datasets. We develop and validate quality control (QC) methods through technical replication, independent samples, comparison to results from fresh-frozen tissue, and recovery of expected associations between gene expression and protein abundance.

Results: Archival tissues from large, multicenter studies showed a much wider range of transcriptional data quality relative to smaller or frozen tissue studies and required stringent QC for subsequent analysis. We developed novel methods for such QC of archival tissue expression profiles based on sample dynamic range and per-study median profile. This enabled validated identification of gene signatures of microsatellite instability and additional features of CRC, and improved recovery of associations between gene expression and protein abundance of MLH1, FASN, CDX2, MGMT, and SIRT1 in CRC tumors.

Conclusions: These methods for large-scale QC of FFPE expression profiles enable study of the cancer transcriptome in relation to extensive clinicopathological information, tumor molecular biomarkers, and long-term lifestyle and outcome data. Clin Cancer Res; 18(22); 6136–46. ©2012 AACR.
Targeted MET Inhibition in Castration-Resistant Prostate Cancer: A Randomized Phase II Study and Biomarker Analysis with Rilotumumab plus Mitoxantrone and Prednisone
Authors: Ryan Charles J. , Rosenthal Mark , Ng Siobhan , Alumkal Joshi , Picus Joel , Gravis Gwenaëlle , Fizazi Karim , Forget Frédéric , Machiels Jean-Pascal , Srinivas Sandy , Zhu Min , Tang Rui , Oliner Kelly S. , Jiang Yizhou , Loh Elwyn , Dubey Sarita , Gerritsen Winald R.
Published: November 2012

Purpose: To evaluate the efficacy, safety, biomarkers, and pharmacokinetics of rilotumumab, a fully human, monoclonal antibody against hepatocyte growth factor (HGF)/scatter factor, combined with mitoxantrone and prednisone (MP) in patients with castration-resistant prostate cancer (CRPC).

Experimental Design: This double-blinded phase II study randomized (1:1:1) patients with progressive, taxane-refractory CRPC to receive MP (12 mg/m2 i.v. day 1, 5 mg twice a day orally days 1–21, respectively) plus 15 mg/kg rilotumumab, 7.5 mg/kg rilotumumab, or placebo (i.v. day 1) every 3 weeks. The primary endpoint was overall survival (OS).

Results: One hundred and forty-four patients were randomized. Median OS was 12.2 versus 11.1 months [HR, 1.10; 80% confidence interval (CI), 0.82–1.48] in the combined rilotumumab versus control arms. Median progression-free survival was 3.0 versus 2.9 months (HR, 1.02; 80% CI, 0.79–1.31). Treatment appeared well tolerated with peripheral edema (24% vs. 8%) being more common with rilotumumab. A trend toward unfavorable OS was observed in patients with high tumor MET expression regardless of treatment. Soluble MET levels increased in all treatment arms. Total HGF levels increased in the rilotumumab arms. Rilotumumab showed linear pharmacokinetics when co-administered with MP.

Conclusions: Rilotumumab plus MP had manageable toxicities and showed no efficacy improvements in this estimation study. High tumor MET expression may identify patients with CRPC with poorer prognosis. Clin Cancer Res; 19(1); 215–24. ©2012 AACR.
Is There a Link Between Genome-Wide Hypomethylation in Blood and Cancer Risk?
Authors: Brennan Kevin , Flanagan James M.
Published: November 2012

Cancer cells display widespread genetic and epigenetic abnormalities, but the contribution to disease risk, particularly in normal tissue before disease, is not yet established. Genome-wide hypomethylation occurs frequently in tumors and may facilitate chromosome instability, aberrant transcription and transposable elements reactivation. Several epidemiologic case–control studies have reported genomic hypomethylation in peripheral blood of cancer patients, suggesting a systemic effect of hypomethylation on disease predisposition, which may be exploited for biomarker development. However, more recent studies have failed to reproduce this. Here, we report a meta-analysis, indicating a consistent inverse association between genomic 5-methylcytosine levels and cancer risk [95% confidence interval (CI), 1.2–6.1], but no overall risk association for studies using surrogates for genomic methylation, including methylation at the LINE-1 repetitive element (95% CI, 0.8–1.7). However, studies have been highly heterogeneous in terms of experimental design, assay type, and analytical methods. We discuss the limitations of the current approaches, including the low interindividual variability of surrogate assays such as LINE1 and the importance of using prospective studies to investigate DNA methylation in disease risk. Insights into genomic location of hypomethylation, from recent whole genome, high-resolution methylome maps, will help address this interesting and clinically important question. Cancer Prev Res; 5(12); 1345–57. ©2012 AACR.
Sleepiness at the Time of Testing Impairs Olfactory Performance
Authors: Ghielmini Enea , Poryazova Rositsa , Baumann Christian R. , Bassetti Claudio L.
Published: November 2012

Background: We aimed to investigate the differential roles of hypocretin versus dopamine dysfunction versus excessive daytime sleepiness (EDS) in the development of hyposmia. Olfaction in patients with Parkinson disease, restless leg syndrome, narcolepsy with cataplexy, EDS, and healthy controls was compared. Methods: Sixty-six subjects participated in the study: 14 with PD, 13 with NC, 12 with RLS, 8 with EDS, and 20 healthy controls. Olfaction was tested using standardized Sniffin’Sticks test. Sleepiness was assessed using Karolinska, Stanford and Epworth sleepiness scales. Results: Olfactory discrimination correlated negatively with subjective momentary sleepiness. A significant deficit in olfaction was found in PD patients with respect to all other groups in all olfactory domains. No significant differences were found between the other groups. Conclusion: We could not confirm decreased olfaction in patients with NC. Yet the significant correlation between momentary sleepiness and olfactory function suggests that sleepiness is a potential confounding factor in the assessment of olfaction, e.g. in NC. Furthermore, our results confirm that olfaction is impaired in PD, whereas it is normal in RLS.
Arrhythmogenic Right Ventricular Cardiomyopathy: The Challenge of Genetic Interpretation in Clinically Suspected Cases
Authors: Anastasakis Aris , Vouliotis Apostolos-Ilias , Protonotarios Nikos , Stefanadis Christodoulos
Published: November 2012

This is the case of a 43-year-old Caucasian man with frequent episodes of paroxysmal atrial fibrillation (AF) and normal resting electrocardiogram (ECG), who fulfilled two minor diagnostic criteria for arrhythmogenic right ventricular cardiomyopathy (ARVC): late potentials by signal-averaged ECG and regional right ventricular outflow tract (RVOT) dyskinesia with mildly dilated RVOT end-diastolic diameter. Genetic test results revealed a disease-associated missense mutation in DSC2 (p.E102K), adding a major diagnostic criterion according to recently published modified Task Force Criteria. However, 2 years after successful ablative therapy for AF, the patient remains completely asymptomatic, without any clinical signs of ARVC. Both ventricular and supraventricular arrhythmias had vanished after AF ablation. Our patient mainly suffered AF without significant ventricular arrhythmias, a very uncommon clinical presentation of ARVC.
A Beautiful Stroke? A Side Note on the 75th Anniversary of the Spectacular Death of the French Organist and Composer Louis Vierne (1870–1937)
Authors: Foerch Christian , Lemercier Sophie , Hennerici Michael G.
Published: November 2012

The great French organist and composer Louis Vierne (1870–1937) died while performing an organ recital at Notre Dame cathedral in Paris – right in front of the console. This historical article provides insights into the biography of a highly talented musician who was challenged by disability and diseases throughout his career. A special focus is placed on the circumstances of Vierne’s remarkable death. Until now, both a primary cerebrovascular event and a ‘heart attack’ are discussed in reference books and encyclopedias as the immanent causes of death. From the perspective of a stroke neurologist, a reappraisal of Vierne’s medical history and the events that happened during his last concert is presented.
Fluorescence Imaging of Intracellular Calcium Signals in Intact Kidney Tissue
Authors: Peppiatt-Wildman C.M. , Crawford C. , Hall A.M.
Published: November 2012

Background: Intracellular calcium (Ca2+) plays an important role in normal renal physiology and in the pathogenesis of various kidney diseases; however, the study of Ca2+ signals in intact tissue has been limited by technical difficulties, including achieving adequate loading of Ca2+-sensitive fluorescent dyes. The kidney slice preparation represents a model whereby three-dimensional tissue architecture is preserved and structures in both the cortex and medulla can be imaged using confocal or multiphoton microscopy. Methods: Ca2+-sensitive dyes Rhod-2, Fura-red and Fluo-4 were loaded into tubular and vascular cells in rat kidney slices using a re-circulating perfusion system and real-time imaging of Ca2+ signals was recorded by confocal microscopy. Kidney slices were also obtained from transgenic mice expressing the GCaMP2 Ca2+-sensor in their endothelial cells and real time Ca2+ transients stimulated by physiological stimuli. Results: Wide spread loading of Ca2+ indicators was achieved in the tubular and vascular structures of both the medulla and cortex. Real time Ca2+ signals were successfully recorded in different intracellular compartments of both rat and mouse cortical and medullary tubules in response to physiological stimuli (ATP and angiotensin II). Glomerular Ca2+ transients were similarly recorded in kidney slices taken from the transgenic mouse expressing the GCaMP2 Ca2+-sensor. Conclusion: We present new approaches that can be adopted to image cytosolic and mitochondrial Ca2+ signals within various cell types in intact kidney tissue. Moreover, techniques described in this study can be used to facilitate future detailed investigations of intracellular Ca2+ homeostasis in renal health and disease.
Alzheimer’s Disease: Differences of Transdermal versus Oral Treatment on Caregiving Time
Authors: Riedel O. , Emmrich A. , Klotsche J. , Dodel R. , Förstl H. , Maier W. , Reichmann H. , Wittchen H.-U.
Published: November 2012

Background: Data on indirect effects of dementia treatment on caregiver burden obtained from naturalistic studies are still lacking. We explored differences between patients with oral and transdermal application of acetylcholine esterase inhibitors regarding caregiver’s time burden and psychopathology. Methods: A cross-sectional naturalistic cohort study of 403 patients in outpatient care with three treatment groups (none, oral, and transdermal) was conducted. Assessments included a standardized clinical burden questionnaire and a standardized caregiver interview. Results: Any treatment was associated with lower burden in most measures. Transdermal treatment was superior regarding (1) administration time (p < 0.001); (2) rates of administration problems (p = 0.031); (3) burden in activities of daily living (p = 0.008), and (4) caregiver anxiety (OR 0.25; 95% CI 0.05–0.99). Caregivers did not report better quality of life regarding mental/physical health. Physicians’ and caregivers’ ratings of patients’ improvements were not associated (ĸ = 0.01–0.06). Conclusions: Benefits associated with transdermal treatment do not translate into a better ‘generic quality of life’ of the caregiver. The substantially different perceptions of patients’ improvements need to be considered in future studies.
Pemetrexed for Previously Treated Patients with Non-Small Cell Lung Cancer and Differences in Efficacy according to Thymidylate Synthase Expression
Authors: Igawa Satoshi , Ryuge Shinichiro , Wada Mayuko , Otani Sakiko , Maki Sachiyo , Takakura Akira , Katono Ken , Sasaki Jiichiro , Sato Yuichi , Masuda Noriyuki
Published: November 2012

The purpose of this study was to evaluate the efficacy of pemetrexed monotherapy in previously treated patients with advanced non-small cell lung cancer (NSCLC) including salvage treatment, and to evaluate whether thymidylate synthase (TS) expression is a predictor for pemetrexed efficacy. Hundred and four previously treated patients with advanced NSCLC who received pemetrexed monotherapy were retrospectively evaluated for clinical efficacy and toxicity. If available, tissue specimens of patients were also analyzed immunohistochemically for TS expression. The patients’ median age was 65 years (range: 43–82). An overall response rate of 9.6% and a median progression-free survival (PFS) time of 3.4 months were achieved. The response rates for the second-line, third-line, fourth-line or further treatments were 9.1, 9.3 and 10.2% (p = 0.33); the median PFS were 3.3, 3.2 and 3.8 months (p = 0.21). The median follow-up duration was 14.9 months; the median overall survival (OS) was 11.9 months. The median PFS and OS were significantly longer in the TS-negative group than in the TS-positive group (5.8 months vs. 1.6 months; p = 0.03, and 14.7 months vs. 8.6 months; p = 0.04, respectively). Pemetrexed monotherapy could be considered as an option in the fourth or later lines of treatment of previously treated patients with advanced NSCLC as well as a second- or third-line treatment, and TS expression may be a potentially predictive factor for pemetrexed efficacy in NSCLC patients.
Is Peak Concentration Needed in Therapeutic Drug Monitoring of Vancomycin? A Pharmacokinetic-Pharmacodynamic Analysis in Patients with Methicillin-Resistant Staphylococcus aureus Pneumonia
Authors: Suzuki Yosuke , Kawasaki Kanako , Sato Yuhki , Tokimatsu Issei , Itoh Hiroki , Hiramatsu Kazufumi , Takeyama Masaharu , Kadota Jun-ichi
Published: November 2012

Background: We analyzed the pharmacokinetic-pharmacodynamic relationship of vancomycin to determine the drug exposure parameters that correlate with the efficacy and nephrotoxicity of vancomycin in patients with methicillin-resistant Staphylococcus aureus pneumonia and evaluated the need to use peak concentration in therapeutic drug monitoring (TDM). Methods: Serum drug concentrations of 31 hospitalized patients treated with vancomycin for methicillin-resistant S. aureus pneumonia were collected. Results: Significant differences in trough concentration (Cmin)/minimum inhibitory concentration (MIC) and area under the serum concentration-time curve (AUC0–24)/MIC were observed between the response and non-response groups. Significant differences in Cmin and AUC0–24 were observed between the nephrotoxicity and non-nephrotoxicity groups. Receiver operating characteristic curves revealed high predictive values of Cmin/MIC and AUC0–24/MIC for efficacy and of Cmin and AUC0–24 for safety of vancomycin. Conclusions: These results suggest little need to use peak concentration in vancomycin TDM because Cmin/MIC and Cmin are sufficient to predict the efficacy and safety of vancomycin.