bluuebunny/crossref_metadata_2025 huggingface dataset:
Perturbations in intracellular Ca2+ handling in skeletal muscle in the G93A*SOD1 mouse model of amyotrophic lateral sclerosis
Authors: Chin Eva R. , Chen Dapeng , Bobyk Kostyantyn D. , Mázala Davi A. G.
Published: September 2014

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by skeletal muscle atrophy and weakness, ultimately leading to respiratory failure. The purpose of this study was to assess changes in skeletal muscle excitation-contraction (E-C) coupling and intracellular Ca2+ handling during disease progression in the G93A*SOD1 ALS transgenic (ALS Tg) mouse model. To assess E-C coupling, single muscle fibers were electrically stimulated (10–150 Hz), and intracellular free Ca2+ concentration was assessed using fura-2. There were no differences in peak fura-2 ratio at any stimulation frequency at 70 days (early presymptomatic). However, at 90 days (late presymptomatic) and 120–140 days (symptomatic), fura-2 ratio was increased at 10 Hz in ALS Tg compared with wild-type (WT) fibers (0.670 ± 0.02 vs. 0.585 ± 0.02 for 120–140 days; P < 0.05). There was also a significant increase in resting fura-2 ratio at 90 days (0.351 ± 0.008 vs. 0.390 ± 0.009 in WT vs. ALS Tg; P < 0.05) and 120–140 days (0.374 ± 0.001 vs. 0.415 ± 0.003 in WT vs. ALS Tg; P < 0.05). These increases in intracellular Ca2+ in ALS Tg muscle were associated with reductions in the sarcoplasmic/endoplasmic reticulum Ca2+ ATPase proteins SERCA1 (to 54% and 19% of WT) and SERCA2 (to 56% and 11% of WT) and parvalbumin (to 80 and 62% of WT) in gastrocnemius muscle at 90 and 120–140 days, respectively. There was no change in dihydropyridine receptor/l-type Ca2+ channel at any age. Overall, these data demonstrate minimal changes in electrically evoked Ca2+ transients but elevations in intracellular Ca2+ attributable to decreased Ca2+-clearance proteins. These data suggest that elevations in cellular Ca2+ could contribute to muscle weakness during disease progression in ALS mice.
Functional mitochondrial analysis in acute brain sections from adult rats reveals mitochondrial dysfunction in a rat model of migraine
Authors: Fried Nathan T. , Moffat Cynthia , Seifert Erin L. , Oshinsky Michael L.
Published: September 2014

Mitochondrial dysfunction has been implicated in many neurological disorders that only develop or are much more severe in adults, yet no methodology exists that allows for medium-throughput functional mitochondrial analysis of brain sections from adult animals. We developed a technique for quantifying mitochondrial respiration in acutely isolated adult rat brain sections with the Seahorse XF Analyzer. Evaluating a range of conditions made quantifying mitochondrial function from acutely derived adult brain sections from the cortex, cerebellum, and trigeminal nucleus caudalis possible. Optimization of this technique demonstrated that the ideal section size was 1 mm wide. We found that sectioning brains at physiological temperatures was necessary for consistent metabolic analysis of trigeminal nucleus caudalis sections. Oxygen consumption in these sections was highly coupled to ATP synthesis, had robust spare respiratory capacities, and had limited nonmitochondrial respiration, all indicative of healthy tissue. We demonstrate the effectiveness of this technique by identifying a decreased spare respiratory capacity in the trigeminal nucleus caudalis of a rat model of chronic migraine, a neurological disorder that has been associated with mitochondrial dysfunction. This technique allows for 24 acutely isolated sections from multiple brain regions of a single adult rat to be analyzed simultaneously with four sequential drug treatments, greatly advancing the ability to study mitochondrial physiology in adult neurological disorders.
Characterization of duodenal expression and localization of fatty acid-sensing receptors in humans: relationships with body mass index
Authors: Little Tanya J. , Isaacs Nicole J. , Young Richard L. , Ott Raffael , Nguyen Nam Q. , Rayner Christopher K. , Horowitz Michael , Feinle-Bisset Christine
Published: September 2014

Fatty acids (FAs) stimulate the secretion of gastrointestinal hormones, including cholecystokinin (CCK) and glucagon like peptide-1 (GLP-1), which suppress energy intake. In obesity, gastrointestinal responses to FAs are attenuated. Recent studies have identified a key role for the FA-sensing receptors cluster of differentiation (CD)36, G protein-coupled receptor (GPR)40, GPR120, and GPR119 in mediating gastrointestinal hormone secretion. This study aimed to determine the expression and localization of these receptors in the duodenum of humans and to examine relationships with obesity. Duodenal mucosal biopsies were collected from nine lean [body mass index (BMI): 22 ± 1 kg/m2], six overweight (BMI: 28 ± 1 kg/m2), and seven obese (BMI: 49 ± 5 kg/m2) participants. Absolute levels of receptor transcripts were quantified using RT-PCR, while immunohistochemistry was used for localization. Transcripts were expressed in the duodenum of lean, overweight, and obese individuals with abundance of CD36>>GPR40>GPR120>GPR119. Expression levels of GPR120 ( r = 0.46, P = 0.03) and CD36 ( r = 0.69, P = 0.0004) were directly correlated with BMI. There was an inverse correlation between expression of GPR119 with BMI ( r2 = 0.26, P = 0.016). Immunolabeling studies localized CD36 to the brush border membrane of the duodenal mucosa and GPR40, GPR120, and GPR119 to enteroendocrine cells. The number of cells immunolabeled with CCK ( r = −0.54, P = 0.03) and GLP-1 ( r = −0.49, P = 0.045) was inversely correlated with BMI, such that duodenal CCK and GLP-1 cell density decreased with increasing BMI. In conclusion, CD36, GPR40, GPR120, and GPR119 are expressed in the human duodenum. Transcript levels of duodenal FA receptors and enteroendocrine cell density are altered with increasing BMI, suggesting that these changes may underlie decreased gastrointestinal hormone responses to fat and impaired energy intake regulation in obesity.
Systems biology analysis of omeprazole therapy in cirrhosis demonstrates significant shifts in gut microbiota composition and function
Authors: Bajaj Jasmohan S. , Cox I. Jane , Betrapally Naga S. , Heuman Douglas M. , Schubert Mitchell L. , Ratneswaran Maiyuran , Hylemon Phillip B. , White Melanie B. , Daita Kalyani , Noble Nicole A. , Sikaroodi Masoumeh , Williams Roger , Crossey Mary M. E. , Taylor-Robinson Simon D. , Gillevet Patrick M.
Published: September 2014

Proton pump inhibitors (PPI) have been associated with infectious complications in cirrhosis, but their impact on distal gut microbiota composition and function is unclear. We aimed to evaluate changes in stool microbiota composition and function in patients with cirrhosis and healthy controls after omeprazole therapy. Both 15 compensated cirrhotic patients and 15 age-matched controls underwent serum gastrin measurement, stool microbiota profiling with multitagged pyrosequencing, and urinary metabolic profiling with NMR spectroscopy to assess microbial cometabolites before/after a 14-day course of 40 mg/day omeprazole under constant diet conditions. Results before (pre) and after PPI were compared in both groups, compared with baseline by systems biology techniques. Adherence was >95% without changes in diet or MELD (model for end-stage liver disease) score during the study. Serum gastrin concentrations significantly increased after PPI in cirrhosis (pre 38.3 ± 35.8 vs. 115.6 ± 79.3 pg/ml P < 0.0001) and controls (pre 29.9 ± 14.5 vs. 116.0 ± 74.0 pg/ml, P = 0.001). A significant microbiota change was seen in both controls and cirrhosis after omeprazole (QIIME P < 0.0001). Relative Streptococcaceae abundance, normally abundant in saliva, significantly increased postomeprazole in controls (1 vs. 5%) and cirrhosis (0 vs. 9%) and was correlated with serum gastrin levels ( r = 0.4, P = 0.005). We found significantly reduced hippurate in cirrhosis vs. controls both pre- and postomeprazole and increased lactate in both groups post vs. preomeprazole, whereas dimethylamine (DMA) decreased in cirrhosis only. On correlation network analysis, significant changes in linkages of bacteria with metabolites (hippurate/DMA/lactate) were found postomeprazole, compared with pre-PPI in cirrhosis patients. In conclusion, omeprazole is associated with a microbiota shift and functional change in the distal gut in patients with compensated cirrhosis that could set the stage for bacterial overgrowth.
Overnutrition, mTOR signaling, and cardiovascular diseases
Authors: Jia Guanghong , Aroor Annayya R. , Martinez-Lemus Luis A. , Sowers James R.
Published: September 2014

The prevalence of obesity and associated medical disorders has increased dramatically in the United States and throughout much of the world in the past decade. Obesity, induced by excess intake of carbohydrates and fats, is a major cause of Type 2 diabetes, hypertension, and the cardiorenal metabolic syndrome. There is emerging evidence that excessive nutrient intake promotes signaling through the mammalian target of rapamycin (mTOR), which, in turn, may lead to alterations of cellular metabolic signaling leading to insulin resistance and obesity-related diseases, such as diabetes, cardiovascular and kidney disease, as well as cancer. While the pivotal role of mTOR signaling in regulating metabolic stress, autophagy, and adaptive immune responses has received increasing attention, there remain many gaps in our knowledge regarding this important nutrient sensor. For example, the precise cellular signaling mechanisms linking excessive nutrient intake and enhanced mTOR signaling with increased cardiovascular and kidney disease, as well as cancer, are not well understood. In this review, we focus on the effects that the interaction between excess intake of nutrients and enhanced mTOR signaling have on the promotion of obesity-associated diseases and potential therapeutic strategies involving targeting mTOR signaling.
Gaze locations affect explicit process but not implicit process during visuomotor adaptation
Authors: Rand Miya K. , Rentsch Sebastian
Published: September 2014

The role of vision in implicit and explicit processes involved in adaptation to novel visuomotor transformations is not well-understood. We manipulated subjects' gaze locations through instructions during a visuomotor rotation task that established a conflict between implicit and explicit processes. Subjects were informed of a rotated visual feedback (45° counterclockwise from the desired target) and instructed to counteract it by using an explicit aiming strategy to the neighboring target (45° clockwise from the target). Simultaneously, they were instructed to gaze at either the desired target (target-gaze group), the neighboring target (hand-target-gaze group), or anywhere (free-gaze group) during aiming. After initial elimination of behavioral errors caused by strategic aiming, the subjects gradually overcompensated the rotation in the early practice, thereby increasing behavioral errors (i.e., a drift). This was caused by an implicit adaptation overriding the explicit strategy. Notably, prescribed gaze locations did not affect this implicit adaptation. In the late practice, the target-gaze and free-gaze groups reduced the drift, whereas the hand-target-gaze group did not. Furthermore, the free-gaze group changed gaze locations for strategic aiming through practice from the neighboring target to the desired target. The onset of this change was correlated with the onset of the drift reduction. These results suggest that gaze locations critically affect explicit adjustments of aiming directions to reduce the drift by taking into account the implicit adaptation that is occurring in parallel. Taken together, spatial eye-hand coordination that ties the gaze and the reach target influences the explicit process but not the implicit process.
Spectral sensitivity differences between rhesus monkeys and humans: implications for neurophysiology
Authors: Lindbloom-Brown Zachary , Tait Leah J. , Horwitz Gregory D.
Published: September 2014

Spectral sensitivity of humans and rhesus monkeys was compared using identical displays and similar procedures. Detection thresholds were measured for the following: 1) 15-Hz modulation of a blue and a green cathode-ray tube phosphor; 2) 15-Hz modulation of all three phosphors together; and 3) slow (<1 Hz) modulations of a blue and a green phosphor under scotopic conditions. Monkeys had lower blue-to-green threshold ratios than humans at all eccentricities tested (0.5 to 7°), consistent with a lower lens optical density in monkeys. In addition to apparently having a lower lens density than humans, monkeys were more sensitive to 15-Hz red-green isoluminant modulations than humans, an effect that cannot be explained by optical factors.
An Implementation Model of Teaching Evaluation Questionnaire System Based on Cloud Computing
Authors: Chen Chih-Yung , Tu Jih-Fu
Published: September 2014

The teaching evaluation questionnaire in the university has been an essential routine. The administrative staff of academic affairs shall obey the rules, fairness, and valid principles within the period of finishing teaching evaluation questionnaire. To ensure the accuracy and timeliness of the large number of questionnaires, it makes the administrative staff of academic affairs face the challenge and pressure if the students are too many. Program potency low or bad often came from the development staff being familiar with bad program development idea; the bad designs of system program in itself causes the factor proportion to occupy more higher, because the similar application program mode of writing is, respectively, different. However, the exploiter may find the method of the little consumption of system resources to complete the similar work task. We develop the software service of cloud platform and the educational administration personnel operates process in the software service of cloud platform. The cloud platform reduces the system wait and calculation time of questionnaire, improves and simplifies system operation flow, and promotes 10 times of program efficiencies as well as solving the load overweight problem of potency bottleneck.
Cytolethal Distending Toxin: A Unique Variation on the AB Toxin Paradigm
Authors: DiRienzo Joseph M.
Published: September 2014

Some of the most potent toxins produced by plants and bacteria are members of a large family known as the AB toxins. AB toxins are generally characterized by a heterogenous complex consisting of two protein chains arranged in various monomeric or polymeric configurations. The newest class within this superfamily is the cytolethal distending toxin (Cdt). The Cdt is represented by a subfamily of toxins produced by a group of taxonomically distinct Gram negative bacteria. Members of this subfamily have a related AB-type chain or subunit configuration and properties distinctive to the AB paradigm. In this review, the unique structural and cytotoxic properties of the Cdt subfamily, target cell specificities, intoxication pathway, modes of action, and relationship to the AB toxin superfamily are compared and contrasted.
Design Optimization of Mechanical Components Using an Enhanced Teaching‐Learning Based Optimization Algorithm with Differential Operator
Authors: Thamaraikannan B. , Thirunavukkarasu V.
Published: September 2014

This paper studies in detail the background and implementation of a teaching‐learning based optimization (TLBO) algorithm with differential operator for optimization task of a few mechanical components, which are essential for most of the mechanical engineering applications. Like most of the other heuristic techniques, TLBO is also a population‐based method and uses a population of solutions to proceed to the global solution. A differential operator is incorporated into the TLBO for effective search of better solutions. To validate the effectiveness of the proposed method, three typical optimization problems are considered in this research: firstly, to optimize the weight in a belt‐pulley drive, secondly, to optimize the volume in a closed coil helical spring, and finally to optimize the weight in a hollow shaft. have been demonstrated. Simulation result on the optimization (mechanical components) problems reveals the ability of the proposed methodology to find better optimal solutions compared to other optimization algorithms.
Location Prediction‐Based Data Dissemination Using Swarm Intelligence in Opportunistic Cognitive Networks
Authors: Li Jie , Wang Xingwei , Jia Jie , Wang Pengfei , Zhou Yan , Zhao Zhijie
Published: September 2014

Swarm intelligence is widely used in the application of communication networks. In this paper we adopt a biologically inspired strategy to investigate the data dissemination problem in the opportunistic cognitive networks (OCNs). We model the system as a centralized and distributed hybrid system including a location prediction server and a pervasive environment deploying the large‐scale human‐centric devices. To exploit such environment, data gathering and dissemination are fundamentally based on the contact opportunities. To tackle the lack of contemporaneous end‐to‐end connectivity in opportunistic networks, we apply ant colony optimization as a cognitive heuristic technology to formulate a self‐adaptive dissemination‐based routing scheme in opportunistic cognitive networks. This routing strategy has attempted to find the most appropriate nodes conveying messages to the destination node based on the location prediction information and intimacy between nodes, which uses the online unsupervised learning on geographical locations and the biologically inspired algorithm on the relationship of nodes to estimate the delivery probability. Extensive simulation is carried out on the real‐world traces to evaluate the accuracy of the location prediction and the proposed scheme in terms of transmission cost, delivery ratio, average hops, and delivery latency, which achieves better routing performances compared to the typical routing schemes in OCNs.
Safety Impacts of Push‐Button and Countdown Timer on Nonmotorized Traffic at Intersections
Authors: Zhou Bei , Roshandeh Arash M. , Zhang Shengrui
Published: September 2014

This paper applies the random parameters negative binominal model to investigate safety impacts of push‐button and countdown timer on pedestrians and cyclists at urban intersections. To account for possible unobserved heterogeneity which could vary from one intersection to another, random parameters model is introduced. A simulation‐based maximum likelihood method using Halton draws is applied to estimate the maximum likelihood of random parameters in the model. Dataset containing pedestrians’ and cyclists’ crash data of 1,001 intersections from Chicago is utilized to establish the statistical relationship between crash frequencies and potential impact factors. LIMDEP (Version 9.0) statistical package is utilized for modeling. The parameter estimation results indicate that existence of push‐button and countdown timer could significantly reduce crash frequencies of pedestrians and cyclists at intersections. Increasing number of through traffic lanes, left turn lanes, and ratio of major direction AADT to minor direction AADT, tend to increase crash frequencies. Annual average daily left turn traffic has a negative impact on pedestrians’ safety, but its impact on cyclists’ crash frequency is statistically insignificant at 90% confidence level. The results of current study could provide important insights for nonmotorized traffic safety improvement projects in both planning and operational levels.
Effects of Processing (Boiling and Roasting) on the Nutritional and Antinutritional Properties of Bambara Groundnuts (Vigna subterranea [L.] Verdc.) from Southern Kaduna, Nigeria
Authors: Ndidi Uche Samuel , Ndidi Charity Unekwuojo , Aimola Idowu Asegame , Bassa Obed Yakubu , Mankilik Mary , Adamu Zainab
Published: September 2014

This research analyzed the effect of processing (boiling and roasting) on the proximate, antinutrient, and mineral composition of Vigna subterranea seeds. The proximate composition analysis showed significant difference (P<0.05) between the levels of crude lipid, crude fiber, gross energy, carbohydrate, and moisture content in the raw and processed V. subterranea. However, no significant difference (P<0.05) was observed in protein content of processed V. subterranea as compared to the raw seeds. Analyses of antinutrient composition show that processing significantly reduced the levels of oxalate, tannins, phytate, trypsin inhibitor, and hydrogen cyanide contents of V. subterranea. While magnesium, potassium, and phosphorus were the most abundant macrominerals in V. subterranea, zinc was the most abundant micromineral. Correlation analysis revealed that the levels of crude protein, ash, moisture, and lipid were negatively affected by phytate, tannins, and oxalate. The results, therefore, suggest that processing significantly lowered the levels of antinutrients in V. subterranea, thereby making it safer for consumption.
A Case of Onychomycosis Caused byRhodotorula glutinis
Authors: Uludag Altun Hatice , Meral Tuba , Turk Aribas Emel , Gorpelioglu Canan , Karabicak Nilgun
Published: September 2014

Rhodotorulaspp. have emerged as opportunistic pathogens, particularly in immunocompromised patients. The current study reports a case of onychomycosis caused byRhodotorula glutinisin a 74-year-old immunocompetent female. The causative agent was identified asR. glutinisbased on the pinkish-orange color; mucoid-appearing yeast colonies on Sabouraud Dextrose Agar at 25°C; morphological evaluation in the Corn Meal-Tween 80 agar; observed oval/round budding yeast at 25°C for 72 hours; no observed pseudohyphae; positive urease activity at 25°C for 4 days; and assimilation features detected by API ID 32C kit and automated Vitek Yeast Biochemical Card 2 system. Antifungal susceptibility test results were as follows: amphotericin B (MIC = 0.5 µg/mL), fluconazole (MIC = 128 µg/mL), itraconazole (MIC = 0.125 µg/mL), voriconazole (MIC = 1 µg/mL), posaconazole (MIC = 0.5 µg/mL), anidulafungin (MIC = 0.5 µg/mL), and caspofungin (MIC = 16 µg/mL). Antifungal therapy was initiated with oral itraconazole at a dose of 400 mg/day; seven-day pulse therapy was planned at intervals of three weeks. Clinical recovery was observed in the clinical evaluation of the patient before the start of the third cure. AlthoughR. glutinishas rarely been reported as the causative agent of onychomycosis, it should be considered.
The Preventable Risk Integrated ModEl and Its Use to Estimate the Health Impact of Public Health Policy Scenarios
Authors: Scarborough Peter , Harrington Richard A. , Mizdrak Anja , Zhou Lijuan Marissa , Doherty Aiden
Published: September 2014

Noncommunicable disease (NCD) scenario models are an essential part of the public health toolkit, allowing for an estimate of the health impact of population-level interventions that are not amenable to assessment by standard epidemiological study designs (e.g., health-related food taxes and physical infrastructure projects) and extrapolating results from small samples to the whole population. The PRIME (Preventable Risk Integrated ModEl) is an openly available NCD scenario model that estimates the effect of population-level changes in diet, physical activity, and alcohol and tobacco consumption on NCD mortality. The structure and methods employed in the PRIME are described here in detail, including the development of open source code that will support a PRIME web application to be launched in 2015. This paper reviews scenario results from eleven papers that have used the PRIME, including estimates of the impact of achieving government recommendations for healthy diets, health-related food taxes and subsidies, and low-carbon diets. Future challenges for NCD scenario modelling, including the need for more comparisons between models and the improvement of future prediction of NCD rates, are also discussed.
Ag-Presenting CpG-Activated pDCs Prime Th17 Cells That Induce Tumor Regression
Authors: Guéry Leslie , Dubrot Juan , Lippens Carla , Brighouse Dale , Malinge Pauline , Irla Magali , Pot Caroline , Reith Walter , Waldburger Jean-Marc , Hugues Stéphanie
Published: September 2014

Plasmacytoid dendritic cells (pDC) rapidly and massively produce type I IFN and other inflammatory cytokines in response to foreign nucleic acids, thereby indirectly influencing T-cell responses. Moreover, antigen (Ag)-presenting pDCs directly regulate T-cell differentiation. Depending on the immune environment, pDCs exhibit either tolerogenic or immunogenic properties. Here, we show that CpG-activated pDCs promote efficient Th17 differentiation. Indeed, Th17 responses are defective in mice selectively lacking MHCII on pDCs upon antigenic challenge. Importantly, in those mice, the frequency of Th17 cells infiltrating solid tumors is impaired. As a result, the recruitment of infiltrating leukocytes in tumors, including tumor-specific cytotoxic T lymphocytes (CTL), is altered and results in increased tumor growth. Importantly, following immunization with tumor Ag and CpG-B, MHCII-restricted Ag presentation by pDCs promotes the differentiation of antitumor Th17 cells that induce intratumor CTL recruitment and subsequent regression of established tumors. Our results highlight a new role for Ag presenting activated pDCs in promoting the development of Th17 cells and impacting on antitumor immunity. Cancer Res; 74(22); 6430–40. ©2014 AACR.
SA-4-1BBL and Monophosphoryl Lipid A Constitute an Efficacious Combination Adjuvant for Cancer Vaccines
Authors: Srivastava Abhishek K. , Dinc Gunes , Sharma Rajesh K. , Yolcu Esma S. , Zhao Hong , Shirwan Haval
Published: September 2014

Vaccines based on tumor-associated antigens (TAA) have limited therapeutic efficacy due to their weak immunogenic nature and the various immune evasion mechanisms active in advanced tumors. In an effort to overcome these limitations, we evaluated a combination of the T-cell costimulatory molecule SA-4-1BBL with the TLR4 agonist monophosphoryl lipid A (MPL) as a novel vaccine adjuvant system. In the TC-1 mouse allograft model of human papilloma virus (HPV)-induced cancer, a single administration of this combination adjuvant with HPV E7 protein caused tumor rejection in all tumor-bearing mice. On its own, SA-4-1BBL outperformed MPL in this setting. Against established tumors, two vaccinations were sufficient to elicit rejection in the majority of mice. In the metastatic model of Lewis lung carcinoma, vaccination of the TAA survivin with SA-4-1BBL/MPL yielded superior efficacy against pulmonary metastases. Therapeutic efficacy of SA-4-1BBL/MPL was achieved in the absence of detectable toxicity, correlating with enhanced dendritic cell activation, CD8+ T-cell function, and an increased intratumoral ratio of CD8+ T effector cells to CD4+FoxP3+ T regulatory cells. Unexpectedly, use of MPL on its own was associated with unfavorable intratumoral ratios of these T-cell populations, resulting in suboptimal efficacy. The efficacy of MPL monotherapy was restored by depletion of T regulatory cells, whereas eliminating CD8+ T cells abolished the efficacy of its combination with SA-4-1BBL. Mechanistic investigations showed that IFNγ played a critical role in supporting the therapeutic effect of SA-4-1BBL/MPL. Taken together, our results offer a preclinical proof of concept for the use of a powerful new adjuvant system for TAA-based cancer vaccines. Cancer Res; 74(22); 6441–51. ©2014 AACR.
LIM and SH3 Protein 1 Induces TGFβ-Mediated Epithelial–Mesenchymal Transition in Human Colorectal Cancer by Regulating S100A4 Expression
Authors: Wang Hui , Shi Jiaolong , Luo Yuhao , Liao Qing , Niu Ya , Zhang Feifei , Shao Ziyun , Ding Yanqing , Zhao Liang
Published: September 2014

Purpose: The expression of LIM and SH3 protein 1 (LASP1) was upregulated in colorectal cancer cases, thereby contributing to the aggressive phenotypes of colorectal cancer cells. However, we still cannot decipher the underlying molecular mechanism associated with colorectal cancer metastasis.

Experimental Design: In this study, IHC was performed to investigate the expression of proteins in human colorectal cancer tissues. Western blot analysis was used to assess the LASP1-induced signal pathway. Two-dimensional difference gel electrophoresis was performed to screen LASP1-modulated proteins and uncover the molecular mechanism of LASP1. TGFβ was used to induce an epithelial–mesenchymal transition (EMT).

Results: LASP1 expression was correlated with the mesenchymal marker vimentin and was inversely correlated with epithelial markers, namely, E-cadherin and β-catenin, in clinical colorectal cancer samples. The gain- and loss-of-function assay showed that LASP1 induces EMT-like phenotypes in vitro and in vivo. S100A4, identified as a LASP1-modulated protein, was upregulated by LASP1. Moreover, it is frequently coexpressed with LASP1 in colorectal cancer. S100A4 was required for EMT, and an increased cell invasiveness of colorectal cancer cell is induced by LASP1. Furthermore, the stimulation of TGFβ resulted in an activated Smad pathway that increased the expression of LASP1 and S100A4. The depletion of LASP1 or S100A4 expression inhibited the TGFβ signaling pathway. Moreover, it significantly weakened the proinvasive effects of TGFβ on colorectal cancer cells.

Conclusion: These findings elucidate the central role of LASP1 in the TGFβ-mediated EMT process and suggest a potential target for the clinical intervention in patients with advanced colorectal cancer. Clin Cancer Res; 20(22); 5835–47. ©2014 AACR.
SGN–LIV1A: A Novel Antibody–Drug Conjugate Targeting LIV-1 for the Treatment of Metastatic Breast Cancer
Authors: Sussman Django , Smith Leia M. , Anderson Martha E. , Duniho Steve , Hunter Joshua H. , Kostner Heather , Miyamoto Jamie B. , Nesterova Albina , Westendorf Lori , Van Epps Heather A. , Whiting Nancy , Benjamin Dennis R.
Published: September 2014

In this article, we describe a novel antibody–drug conjugate (ADC; SGN–LIV1A), targeting the zinc transporter LIV-1 (SLC39A6) for the treatment of metastatic breast cancer. LIV-1 was previously known to be expressed by estrogen receptor–positive breast cancers. In this study, we show that LIV-1 expression is maintained after hormonal therapy in primary and metastatic sites and is also upregulated in triple-negative breast cancers. In addition to breast cancer, other indications showing LIV-1 expression include melanoma, prostate, ovarian, and uterine cancer. SGN–LIV1A consists of a humanized antibody conjugated through a proteolytically cleavable linker to monomethyl auristatin E, a potent microtubule-disrupting agent. When bound to surface-expressed LIV-1 on immortalized cell lines, this ADC is internalized and traffics to the lysozome. SGN–LIV1A displays specific in vitro cytotoxic activity against LIV-1–expressing cancer cells. In vitro results are recapitulated in vivo where antitumor activity is demonstrated in tumor models of breast and cervical cancer lineages. These results support the clinical evaluation of SGN–LIV1A as a novel therapeutic agent for patients with LIV-1–expressing cancer. Mol Cancer Ther; 13(12); 2991–3000. ©2014 AACR.
CBS9106-Induced CRM1 Degradation Is Mediated by Cullin Ring Ligase Activity and the Neddylation Pathway
Authors: Saito Naoya , Sakakibara Keiichi , Sato Takuji , Friedman Jonathan M. , Kufe Donald W. , VonHoff Daniel D. , Kawabe Takumi
Published: September 2014

Chromosome region maintenance 1 (CRM1) mediates the nuclear export of proteins and mRNAs, and is overexpressed in various cancers. Recent studies have also reported that CRM1 protein expression is a negative prognostic factor in patients with cancer. Therefore, CRM1 is considered a potential target for anticancer therapy. Our previous study demonstrated that CBS9106, a synthetic small-molecular inhibitor of CRM1, decreases CRM1 protein through proteasomal degradation without affecting CRM1 mRNA levels. However, the mechanism by which CRM1 is degraded is not well understood. Here, we demonstrate a novel signaling pathway that plays an important role in CBS9106-induced CRM1 degradation. We found that MLN4924, a selective inhibitor of NEDD8-activating enzyme (NAE), effectively inhibits cullin neddylation and attenuates CBS9106-induced CRM1 degradation in a time- and dose-dependent manner. MLN4924 also attenuated CBS9106-induced nuclear accumulation of Ran-binding protein 1 (RanBP1), cell growth inhibition, and apoptosis. Furthermore, RNAi-mediated knockdown of neddylation pathway proteins (NEDD8 and UBA3) or cullin ring ligase (CRL) component protein (Rbx1) attenuated CRM1 protein degradation and G1 phase cell-cycle arrest by CBS9106. Knockdown of CSN5 or CAND1 also partially inhibited CBS9106-induced CRM1 degradation. These findings demonstrate that CBS9106-induced CRM1 degradation is conferred by CRL activity involving the neddylation pathway, and that this response to CBS9106 leads to cell growth inhibition and apoptosis. Mol Cancer Ther; 13(12); 3013–23. ©2014 AACR.
Triple-Negative Breast Cancer Patients Treated at MD Anderson Cancer Center in Phase I Trials: Improved Outcomes with Combination Chemotherapy and Targeted Agents
Authors: Ganesan Prasanth , Moulder Stacy , Lee J. Jack , Janku Filip , Valero Vicente , Zinner Ralph G. , Naing Aung , Fu Siqing , Tsimberidou Apostolia M. , Hong David , Stephen Bettzy , Stephens Philip , Yelensky Roman , Meric-Bernstam Funda , Kurzrock Razelle , Wheler Jennifer J.
Published: September 2014

Patients with metastatic triple-negative breast cancer (TNBC) have poor treatment outcomes. We reviewed the electronic records of consecutive patients with metastatic TNBC treated in phase I clinic at MD Anderson Cancer Center (Houston, TX) between Augu st 2005 and May 2012. One hundred and six patients received at least 1 phase I trial. Twelve of 98 evaluable patients (12%) had either complete response (CR; n = 1), partial response (PR; n = 7), or stable disease ≥6 months (SD; n = 4). Patients treated on matched therapy (n = 16) compared with those on nonmatched therapy (n = 90) had improved SD ≥ 6 months/PR/CR (33% vs. 8%; P = 0.018) and longer progression-free survival (PFS; median, 6.4 vs. 1.9 months; P = 0.001). Eleven of 57 evaluable patients (19%) treated with combination chemotherapy and targeted therapy had SD ≥ 6 months/PR/CR versus 1 of 41 evaluable patients (2%) treated on other phase I trials (P = 0.013), and longer PFS (3.0 vs. 1.6 months; P < 0.0001). Patients with molecular alterations in the PI3K/AKT/mTOR pathway treated on matched therapy (n = 16) had improved PFS compared with those with and without molecular alterations treated on nonmatched therapy (n = 27; 6.4 vs. 3.2 months; P = 0.036). On multivariate analysis, improved PFS was associated with treatment with combined chemotherapy and targeted agents (P = 0.0002), ≤2 metastatic sites (P = 0.003), therapy with PI3K/AKT/mTOR inhibitors for those with cognate pathway abnormalities (P = 0.018), and treatment with antiangiogenic agents (P = 0.023). In summary, combinations of chemotherapy and angiogenesis and/or PI3K/AKT/mTOR inhibitors demonstrated improved outcomes in patients with metastatic TNBC. Mol Cancer Ther; 13(12); 3175–84. ©2014 AACR.
MEK Inhibitor for Gastric Cancer with MEK1 Gene Mutations
Authors: Sogabe Shunsuke , Togashi Yosuke , Kato Hiroaki , Kogita Akihiro , Mizukami Takuro , Sakamoto Yoichi , Banno Eri , Terashima Masato , Hayashi Hidetoshi , de Velasco Marco A. , Sakai Kazuko , Fujita Yoshihiko , Tomida Shuta , Yasuda Takushi , Takeyama Yoshifumi , Okuno Kiyotaka , Nishio Kazuto
Published: September 2014

The prognosis for patients with unresectable advanced or recurrent gastric cancer remains poor. The identification of additional oncogenes with influences similar to those of epidermal growth factor receptor gene mutations, upon which the growth of cancer cells is dependent, is needed. In this study, we evaluated sensitivity to MEK inhibitors (GSK1120212 and PD0325901) in several gastric cancer cell lines in vitro and found three poorly differentiated gastric cancer cell lines that were hypersensitive to the inhibitors. The sequence analyses in these three cell lines revealed that one cell line had a novel MEK1 mutation, while the other two had previously reported KRAS and MEK1 mutations, respectively; the gene statuses of the other resistant cell lines were all wild-type. Experiments using MEK1 expression vectors demonstrated that the MEK1 mutations induced the phosphorylation of ERK1/2 and had a transforming potential, enhancing the tumorigenicity. The MEK inhibitor dramatically reduced the phosphorylation of ERK1/2 and induced apoptosis in the cell lines with MEK1 mutations. In vivo, tumor growth was also dramatically decreased by an inhibitor. One of the 46 gastric cancer clinical samples that were examined had a MEK1 mutation; this tumor had a poorly differentiated histology. Considering the addiction of cancer cells to active MEK1 mutations for proliferation, gastric cancer with such oncogenic MEK1 mutations might be suitable for targeted therapy with MEK inhibitors. Mol Cancer Ther; 13(12); 3098–106. ©2014 AACR.
Immune Activation and a 9-Year Ongoing Complete Remission Following CD40 Antibody Therapy and Metastasectomy in a Patient with Metastatic Melanoma
Authors: Bajor David L. , Xu Xiaowei , Torigian Drew A. , Mick Rosemarie , Garcia Laura R. , Richman Lee P. , Desmarais Cindy , Nathanson Katherine L. , Schuchter Lynn M. , Kalos Michael , Vonderheide Robert H.
Published: September 2014

Direct immune activation via agonistic mAbs is a potentially complementary approach to therapeutic blockade of inhibitory immune receptors in cancer. Here, we provide genetic analysis of the immunologic consequences associated with the use of an agonistic CD40 mAb in a patient with metastatic melanoma who responded, underwent a single metastasectomy, and then achieved a complete remission ongoing for more than 9 years after starting therapy. Tumor microenvironment after immunotherapy was associated with proinflammatory modulations and emergence of a de novo T-cell repertoire as detected by next-generation sequencing of T-cell receptors (TCR) in the tumor and blood. The de novo T-cell repertoire identified in the posttreatment metastasectomy sample was also present—and in some cases expanded—in the circulation years after completion of therapy. Comprehensive study of this “exceptional responder” highlights the emerging potential of direct immune agonists in the next wave of cancer immunotherapies and a potential role for TCR deep sequencing in cancer immune assessment. Cancer Immunol Res; 2(11); 1051–8. ©2014 AACR.
Merkel Cell Carcinoma: Differences between Sun-Exposed and Non-Sun-Exposed Variants - A Clinical Analysis of 36 Cases
Authors: Marcoval Joaquim , Ferreres Josep Ramon , Penín Rosa María , Pérez Diana , Viñals Joan Maria
Published: September 2014

Background: There are few studies focusing on the clinical characteristics of Merkel cell carcinoma (MCC). Objective: To retrospectively analyze the clinical features of our patients and their relationship with sun exposure. Methods: Thirty-six patients diagnosed with MCC (20 men and 16 women, mean age 72.08 years) were included in the study. Results: 21 patients developed MCC in sun-exposed skin and 15 patients in non-sun-exposed areas. MCC was >2 cm in 19 cases. Six of the 7 patients who died as a result of MCC had non-sun-exposed tumors. Only tumor size >2 cm significantly influenced survival (p = 0.033). Conclusion: Sun-exposed lesions tended to be <2 cm in diameter and were more common in men, while non-sun-exposed tumors were larger, usually occurring in women and carrying a greater likelihood of death by MCC. Non-sun-exposed tumors usually present as fast-growing, multilobar nodular lesions with a smooth shiny surface.
Fahr's Disease Linked to a Novel SLC20A2 Gene Mutation Manifesting with Dynamic Aphasia
Authors: Brighina Laura , Saracchi Enrico , Ferri Francesca , Gagliardi Monica , Tarantino Patrizia , Morzenti Sabrina , Musarra Monica , Patassini Mirko , Annesi Grazia , Ferrarese Carlo
Published: September 2014

Background: Idiopathic basal ganglia calcification (IBGC), also known as Fahr's disease, is a rare disorder characterized by widespread cerebral calcifications, an autosomal dominant pattern of inheritance and clinical and genetic heterogeneity. The recently identified IBGC gene, SLC20A2, encodes for type III sodium-dependent phosphate transporter 2 and its loss-of-function mutations may lead to the regional accumulation of inorganic phosphate in the brain, causing calcium phosphate deposition. Objective: To describe the clinical, neuroimaging and genetic findings in an Italian family with IBGC. Methods: The family members underwent clinical and radiological examination in order to diagnose IBGC according to standard criteria and screening for SLC20A2 gene mutations. The affected subjects also underwent neuropsychological longitudinal assessments and functional neuroimaging investigations. Results: The 2 affected family members harbored a novel missense mutation, G1618A, in the SLC20A2 gene, leading to gly540-to-arg (G540R) substitution in a highly conserved residue. This is the first SLC20A2 gene mutation associated with familial IBGC reported in the Italian population and is damaging according to all prediction programs. In the index case we observed a fair correlation between cortical areas with no calcifications but with significant hypometabolism at [18F]FDG-PET (inferior frontal premotor cortex) and the neuropsychological picture dominated by dynamic aphasia and buccofacial apraxia. Conclusion: These findings expand the catalog of SLC20A2 mutations identified to date and add dynamic aphasia to the spectrum of neuropsychological deficits reported in IBGC, supporting the use of functional neuroimaging studies for better investigation of genotype-phenotype correlations.
Pregnancy Rate in Endometriosis Patients according to the Severity of the Disease after Using a Combined Approach of Laparoscopy, GnRH Agonist Treatment and in vitro Fertilization
Authors: Sõritsa Deniss , Saare Merli , Laisk-Podar Triin , Peters Maire , Sõritsa Andrei , Matt Kadri , Karro Helle , Salumets Andres
Published: September 2014

Aim: To evaluate the effects of combined treatment approaches on endometriosis-associated infertility in different stages of endometriosis using laparoscopy, gonadotropin-releasing hormone (GnRH) agonist (GnRHa) therapy and in vitro fertilization (IVF). Methods: This retrospective study was carried out on 179 women with surgically confirmed endometriosis. Patients were divided into subgroups: group 1 (stage I-II, n = 121) and group 2 (stage III-IV, n = 58). Patients eligible for IVF, who were found to have adenomyosis or moderate to severe endometriosis, were also given postoperative GnRHa. Pregnancy and delivery rates were cumulatively calculated during 5 years according to the severity of the disease. Results: The overall pregnancy, delivery and miscarriage rates were 66.5, 56.4 and 15.1%, respectively, for all patients following spontaneous and assisted conception. There were no significant differences in reproductive outcomes between the study groups. The pregnancy and delivery rates were also comparable within group 1 between the patients with and without GnRHa treatment. Conclusion: Pregnancy and delivery rates at different stages of endometriosis were not affected by the different approaches used for infertility treatment, with >60 and >50% of patients having conceived and delivered a baby, respectively, in both groups. The usefulness of GnRHa treatment for endometriosis patients with minimal to mild forms is questionable and deserves further studies.
Relax, It's Just Laparoscopy! A Prospective Randomized Trial on Heart Rate Variability of the Surgeon in Robot-Assisted versus Conventional Laparoscopic Cholecystectomy
Authors: Heemskerk Jeroen , Zandbergen H. Reinier , Keet Sander W.M. , Martijnse Ingrid , van Montfort Gust , Peters Rob J.A. , Svircevic Vesna , Bouwman R. Arthur , Baeten Cor G.M.I. , Bouvy Nicole D.
Published: September 2014

Background: Laparoscopic surgery might be beneficial for the patient, but it imposes increased physical and mental strain on the surgeon. Robot-assisted laparoscopic surgery addresses some of the laparoscopic drawbacks and may potentially reduce mental strain. This could reduce the risk of surgeon's fatigue, mishaps and strain-induced illnesses, which may eventually improve the safety of laparoscopic surgical procedures. Methods: To test this hypothesis, a randomized study was performed, comparing both heart rate and heart rate variability (HRV) of the surgeon as a measure of total and mental strain, respectively, during conventional and robot-assisted laparoscopic cholecystectomy. Results: Both heart rate and HRV (the low-frequency band/high-frequency band ratio) were significantly decreased when using robotic assistance. Conclusions: These data suggest the use of the daVinci® Surgical System leads to less physical and mental strain of the surgeon during surgery. However, assessing mental strain by means of HRV is cumbersome since there is no clear cutoff point or scale for maximum tolerated strain levels and its related effects on surgeon's health.
Bilateral Sequential NAION following Cataract Extraction: Case Report and Review of the Literature
Authors: Barequet Dana , Moisseiev Elad , Michaeli Adi , Dotan Gad
Published: September 2014

We report a 64-year-old patient who developed nonarteritic ischemic optic neuropathy (NAION) in both eyes following cataract extraction. The cataract surgeries in both eyes were uneventful and performed within a year, and NAION occurred a few months postoperatively in both eyes. A review of the literature on this rare complication is provided. This case report serves to raise awareness among cataract surgeons about this potential complication associated with irreversible visual loss, and especially about its high risk of bilaterality.
Dihydropyrimidine Dehydrogenase Deficiency in Two Malaysian Siblings with Abnormal MRI Findings
Authors: Chen Bee Chin , Mohd Rawi Rowani , Meinsma Rutger , Meijer Judith , Hennekam Raoul C.M. , van Kuilenburg André B.P.
Published: September 2014

Dihydropyrimidine dehydrogenase (DPD) deficiency is an autosomal recessive disorder of the pyrimidine metabolism. Deficiency of this enzyme leads to an accumulation of thymine and uracil and a deficiency of metabolites distal to the catabolic enzyme. The disorder presents with a wide clinical spectrum, ranging from asymptomatic to severe neurological manifestations, including intellectual disability, seizures, microcephaly, autistic behavior, and eye abnormalities. Here, we report on an 11-year-old Malaysian girl and her 6-year-old brother with DPD deficiency who presented with intellectual disability, microcephaly, and hypotonia. Brain MRI scans showed generalized cerebral and cerebellar atrophy and callosal body dysgenesis in the boy. Urine analysis showed strongly elevated levels of uracil in the girl and boy (571 and 578 mmol/mol creatinine, respectively) and thymine (425 and 427 mmol/mol creatinine, respectively). Sequence analysis of the DPYD gene showed that both siblings were homozygous for the mutation c.1651G>A (pAla551Thr).