bluuebunny/crossref_metadata_2025 huggingface dataset:
FUNGIFORM PAPILLAE HYPERPLASIA (FPH) IDENTIFICATION BY TONGUE TEXTURE ANALYSIS
Authors: HUANG BO , LI NAIMIN
Published: August 2009

Computerized tongue diagnosis can make use of a number of pathological features of the tongue. To date, there have been few computerized applications that focus on the very commonly used and distinctive diagnostic and textural features of the tongue, Fungiform Papillae Hyperplasia (FPH). In this paper, we propose a computer-aided system for identifying the presence or absence of FPH. We first define and partition a region of interest (ROI) for texture acquisition. After preprocessing for detection and removal of reflective points, a set of 2D Gabor filter banks is used to extract and represent textural features. Then, we apply the Linear Discriminant Analysis (LDA) to identify the data sets from the tongue image database. The experimental results reasonably demonstrate the effectiveness of the method described in this paper.
AN EMPIRICAL INVESTIGATION OF LICENSING PROBABILITY AND SECTORAL PATTERN IN TECHNOLOGY TRANSFER
Authors: KIM JUNYOUNG , YOUN JANGHYUK , PARK YONGTAE
Published: August 2009

A technology evaluation model based on the negotiation between a licensor and a licensee is proposed. This model also considered information asymmetry which decreases the probability of technology transfer between the licensor and the licensee. Factors which affect this decrease and sectors which are more exposed to this decrease using the technology evaluation model are explored using the pooled technology transferred cases of NTB (National Technology Bank), Korea. The only explanatory factor which covers all sectors is the "technology regime-based innovation pattern," and the factor named "patent authorization stage" is the most influencing variable within a specific sector. In three sectors of the four "technology regime-based innovation patterns," namely, scale intensive, specialized supplier, and science based, the variation of licensing probability is likely to increase when the "patent authorization stage" increases. Using SOFM and ANOVA, the negotiated result and the value of technology evaluated are likely to increase.
IMAGE FUSION ALGORITHM BASED ON NONSUBSAMPLED CONTOURLET TRANSFORM AND ESTIMATION THEORY
Authors: ZHAO GAO-PENG , BO YU-MING
Published: August 2009

Aimed at the fusion of infrared and visual images, and their application demands, a new image fusion method was proposed based on the nonsubsampled contourlet transform and estimation theory. Firstly, the nonsubsampled contourlet transform was employed to decompose the source images into the low frequency subband coefficient and bandpass directional subband coefficients. Then, for the bandpass directional subband coefficients, the detail coefficients were modeled by the Gaussian mixture distributions and the EM algorithm was used in conjunction with the model to develop an iterative fusion procedure to estimate the model parameters and to produce the fused coefficients; for the fusion of the approximate subband coefficients, the rule was employed based on the energy of the pixel neighboring region. Finally, the fused image was obtained by applying the inverse nonsubsampled contourlet transform. The experimental results showed that the fusion scheme is effective and the fused image is better than that of using the wavelet transform and the contourlet transform.
FEASIBILITY OF THE NON-CODING RNA GRADIENTS IN CELLS
Authors: ZHDANOV VLADIMIR P.
Published: August 2009

In eukaryotic cells, the gene transcription often results in the formation of non-coding RNAs (ncRNAs). The key function of such RNAs is to bind to and modulate the activity of mRNAs and/or proteins. To scrutinize this ncRNA function in a cell, the author (i) proposes a spatio-temporal kinetic model, including ncRNA–protein association and degradation, (ii) derives a criterion of feasibility of the ncRNA gradients, and (iii) shows that this criterion can be satisfied with physically reasonable values of the model parameters. Thus, the ncRNA gradients are feasible. For the ncRNA–mRNA association and degradation, the situation is similar. The likely biological role of such gradients is open for debate.
MONTE CARLO STUDY OF LIPID NANOSCALE ORGANIZATION IN CELL MEMBRANES: TUNING DOMAIN SIZE AND STABILITY NEAR THE PHASE BOUNDARY
Authors: REIGADA RAMON
Published: August 2009

Lipids in cell membranes are organized in cholesterol-rich domains that are involved in many important cellular functions. Depending on the cell state, such structures are believed to develop covering a wide range of submicrometric sizes and different levels of stability. Using a simple Monte Carlo approach I demonstrate that when the membrane lipid mixture approaches a phase boundary, the cell has several mechanisms to tune both the size and stability of these domains. Cholesterol levels variation, presence of ceramides and insertion of proteins provide plausible mechanisms for the control of the nanoscale lipid organization in cell membranes.
Differential effects of insulin deprivation and systemic insulin treatment on plasma protein synthesis in type 1 diabetic people
Authors: Jaleel Abdul , Klaus Katherine A. , Morse Dawn M. , Karakelides Helen , Ward Lawrence E. , Irving Brian A. , Nair K. Sreekumaran
Published: August 2009

It remains to be determined whether systemic insulin replacement normalizes synthesis rates of different plasma proteins and whether there are differential effects on various plasma proteins. We tested a hypothesis that insulin deprivation differentially affects individual plasma protein synthesis and that systemic insulin treatment may not normalize synthesis of all plasma proteins. We measured synthesis rates of 41 plasma proteins in seven each of type 1 diabetic (T1DM) and nondiabetic participants (ND) using [ ring-13C6]phenylalanine as a tracer. T1DM were studied while on chronic insulin treatment and during 8 h insulin deprivation. Insulin treatment normalized glucose levels, but plasma insulin levels were higher during insulin treatment than during insulin deprivation in T1DM and ND. Individual plasma proteins were purified by affinity chromatography and two-dimensional gel electrophoresis. Only 41 protein gel spots from over 300 were chosen based on their protein homogeneity. Insulin deprivation and hyperglycemia either significantly increased ( n = 12) or decreased ( n = 12) synthesis rates of 24 of 41 plasma proteins in T1DM compared with ND. Insulin treatment normalized synthesis rates of 13 of these 24 proteins, which were altered during insulin deprivation. However, insulin treatment significantly altered the synthesis of 14 additional proteins. In conclusion, acute insulin deprivation caused both a decrease and increase in synthesis rates of many plasma proteins with various functions. Moreover, chronic systemic insulin treatment not only did not normalize synthesis of all plasma proteins but also altered synthesis of several additional proteins that were unaltered during insulin deprivation.
Genetic impairment of AMPKα2 signaling does not reduce muscle glucose uptake during treadmill exercise in mice
Authors: Maarbjerg Stine J. , Jørgensen Sebastian B. , Rose Adam J. , Jeppesen Jacob , Jensen Thomas E. , Treebak Jonas T. , Birk Jesper B. , Schjerling Peter , Wojtaszewski Jørgen F. P. , Richter Erik A.
Published: August 2009

Some studies suggest that the 5′-AMP-activated protein kinase (AMPK) is important in regulating muscle glucose uptake in response to intense electrically stimulated contractions. However, it is unknown whether AMPK regulates muscle glucose uptake during in vivo exercise. We studied this in male and female mice overexpressing kinase-dead AMPKα2 (AMPK-KD) in skeletal and heart muscles. Wild-type and AMPK-KD mice were exercised at the same absolute intensity and the same relative intensity (30 and 70% of individual maximal running speed) to correct for reduced exercise capacity of the AMPK-KD mouse. Muscle glucose clearance was measured using 2-deoxy-[3H]glucose as tracer. In wild-type mice, glucose clearance was increased at 30 and 70% of maximal running speed by 40 and 350% in the quadriceps muscle and by 120 and 380% in gastrocnemius muscle, respectively. Glucose clearance was not lower in AMPK-KD muscles compared with wild-type regardless of whether animals were exercised at the same relative or the same absolute intensity. In agreement, surface membrane content of the glucose transporter GLUT4 was increased similarly in AMPK-KD and wild-type muscle in response to running. We also measured signaling of alternative exercise-sensitive pathways that might be compensatorily increased in AMPK-KD muscles. However, increases in phosphorylation of CaMKII, Trisk95, p38 MAPK, and ERK1/2 were not higher in AMPK-KD than in WT muscle. Collectively, these findings suggest that AMPKα2 signaling is not essential in regulating glucose uptake in mouse skeletal muscle during treadmill exercise and that other mechanisms play a central role.
MitoQ administration prevents endotoxin-induced cardiac dysfunction
Authors: Supinski G. S. , Murphy M. P. , Callahan L. A.
Published: August 2009

Sepsis elicits severe alterations in cardiac function, impairing cardiac mitochondrial and pressure-generating capacity. Currently, there are no therapies to prevent sepsis-induced cardiac dysfunction. We tested the hypothesis that administration of a mitochondrially targeted antioxidant, 10-(6′-ubiquinonyl)-decyltriphenylphosphonium (MitoQ), would prevent endotoxin-induced reductions in cardiac mitochondrial and contractile function. Studies were performed on adult rodents ( n = 52) given either saline, endotoxin (8 mg·kg−1·day−1), saline + MitoQ (500 μM), or both endotoxin and MitoQ. At 48 h animals were killed and hearts were removed for determination of either cardiac mitochondrial function (using polarography) or cardiac pressure generation (using the Langendorf technique). We found that endotoxin induced reductions in mitochondrial state 3 respiration rates, the respiratory control ratio, and ATP generation. Moreover, MitoQ administration prevented each of these endotoxin-induced abnormalities, P < 0.001. We also found that endotoxin produced reductions in cardiac pressure-generating capacity, reducing the systolic pressure-diastolic relationship. MitoQ also prevented endotoxin-induced reductions in cardiac pressure generation, P < 0.01. One potential link between mitochondrial and contractile dysfunction is caspase activation; we found that endotoxin increased cardiac levels of active caspases 9 and 3 ( P < 0.001), while MitoQ prevented this increase ( P < 0.01). These data demonstrate that MitoQ is a potent inhibitor of endotoxin-induced mitochondrial and cardiac abnormalities. We speculate that this agent may prove a novel therapy for sepsis-induced cardiac dysfunction.
Uremia impacts renal inflammatory cytokine gene expression in the setting of experimental acute kidney injury
Authors: Zager Richard A. , Johnson Ali C. M. , Lund Steve
Published: August 2009

Inflammatory cytokines are evoked by acute kidney injury (AKI) and may contribute to evolving renal disease. However, the impact of AKI-induced uremia on proinflammatory (e.g., TNF-α, MCP-1, TGF-β1) and anti-inflammatory (e.g., IL-10) cytokine gene expression remains unknown. This study was undertaken to gain some initial insights into this issue. CD-1 mice were subjected to left renal ischemia-reperfusion (I/R) in the absence or presence of uremia (± right ureteral transection). TNF-α, MCP-1, TGF-β1, and IL-10 mRNAs, cytokine protein levels, and RNA polymerase II (Pol II) recruitment to these genes were assessed. Renal cytokine mRNA levels were also contrasted with unilateral vs. bilateral renal parenchymal damage (I/R or ureteral obstruction). Potential effects of uremia on cytokine mRNAs in the absence of parenchymal renal damage [bilateral ureteral transection (BUTx)] were sought. Finally, the impact of simulated in vitro uremia (HK-2 tubular cells exposed to peritoneal dialysate from uremic vs. normal mice) on cytokine mRNA and microRNA profiles was assessed. Uremia blunted TNF-α, MCP-1, and TGF-β1 mRNA increases in all three in vivo parenchymal acute renal failure models. These results were paralleled by reductions in cytokine protein levels and Pol II recruitment to their respective genes. Conversely, uremia increased IL-10 mRNA, both in the presence and absence (BUTx) of parenchymal renal damage. The uremic milieu also suppressed HK-2 cell proinflammatory cytokine mRNA levels and altered the expression of least 69 microRNAs ( P < 0.0001). We conclude that both pro- and anti-inflammatory cytokine gene expressions are influenced by uremia, with a potential predilection toward an anti-inflammatory state. Changes in gene transcription (as reflected by Pol II recruitment), and possible posttranscriptional modifications (known to be induced by microRNAs), are likely involved.
Oculomotor Distraction by Signals Invisible to the Retinotectal and Magnocellular Pathways
Authors: Bompas Aline , Sumner Petroc
Published: August 2009

Irrelevant stimulus onsets interfere with saccade planning to other stimuli, prolonging saccadic latency (the oculomotor distractor effect) or eliciting directional errors (saccadic capture). Such stimulus-driven interference has been associated with the retinotectal pathway, the direct pathway from retina to superior colliculus. Consistent with this theory, the distractor effect has not been found for stimuli visible only to the short-wave cones in the retina (S cones), which are thought not to contribute to the retinotectal pathway. However, S-cone signals are generally slower than luminance signals and such differences in temporal dynamics have not been taken into account when investigating the saccadic distractor effect. Here, by varying the delay between target and distractor, we found that S-cone stimuli do in fact produce a distractor effect, but the optimal delay is generally different from that for luminance distractors. The temporal dynamics of the distractor effect conform to a general framework of saccadic competition that takes sensory transmission time into account. Additionally, we observe that S-cone stimuli are able to produce saccadic capture in our paradigm. We conclude that stimulus-driven oculomotor interference does not rely on the retinotectal pathway, or indeed the magnocellular pathway, which is also blind to our S-cone stimuli.
Odorant Concentration Dependence in Electroolfactograms Recorded From the Human Olfactory Epithelium
Authors: Lapid Hadas , Seo Han-Seok , Schuster Benno , Schneidman Elad , Roth Yehudah , Harel David , Sobel Noam , Hummel Thomas
Published: August 2009

Electroolfactograms (EOGs) are the summated generator potentials of olfactory receptor neurons measured directly from the olfactory epithelium. To validate the sensory origin of the human EOG, we set out to ask whether EOGs measured in humans were odorant concentration dependent. Each of 22 subjects (12 women, mean age = 23.3 yr) was tested with two odorants, either valeric acid and linalool ( n = 12) or isovaleric acid and l-carvone ( n = 10), each delivered at four concentrations diluted with warm (37°C) and humidified (80%) odorless air. In behavior, increased odorant concentration was associated with increased perceived intensity (all F > 5, all P < 0.001). In EOG, increased odorant concentration was associated with increased area under the EOG curve (all F > 8, all P < 0.001). These findings substantiate EOG as a tool for probing olfactory coding directly at the level of olfactory receptor neurons in humans.
Reconstruction for Time‐Domain in vivo EPR 3D Multigradient Oximetric Imaging—A Parallel Processing Perspective
Authors: Dharmaraj Christopher D. , Thadikonda Kishan , Fletcher Anthony R. , Doan Phuc N. , Devasahayam Nallathamby , Matsumoto Shingo , Johnson Calvin A. , Cook John A. , Mitchell James B. , Subramanian Sankaran , Krishna Murali C.
Published: August 2009

Three‐dimensional Oximetric Electron Paramagnetic Resonance Imaging using the Single Point Imaging modality generates unpaired spin density and oxygen images that can readily distinguish between normal and tumor tissues in small animals. It is also possible with fast imaging to track the changes in tissue oxygenation in response to the oxygen content in the breathing air. However, this involves dealing with gigabytes of data for each 3D oximetric imaging experiment involving digital band pass filtering and background noise subtraction, followed by 3D Fourier reconstruction. This process is rather slow in a conventional uniprocessor system. This paper presents a parallelization framework using OpenMP runtime support and parallel MATLAB to execute such computationally intensive programs. The Intel compiler is used to develop a parallel C++ code based on OpenMP. The code is executed on four Dual‐Core AMD Opteron shared memory processors, to reduce the computational burden of the filtration task significantly. The results show that the parallel code for filtration has achieved a speed up factor of 46.66 as against the equivalent serial MATLAB code. In addition, a parallel MATLAB code has been developed to perform 3D Fourier reconstruction. Speedup factors of 4.57 and 4.25 have been achieved during the reconstruction process and oximetry computation, for a data set with 23 × 23 × 23 gradient steps. The execution time has been computed for both the serial and parallel implementations using different dimensions of the data and presented for comparison. The reported system has been designed to be easily accessible even from low‐cost personal computers through local internet (NIHnet). The experimental results demonstrate that the parallel computing provides a source of high computational power to obtain biophysical parameters from 3D EPR oximetric imaging, almost in real‐time.
Sterol Biosynthesis Pathway as Target for Anti-trypanosomatid Drugs
Authors: de Souza Wanderley , Rodrigues Juliany Cola Fernandes
Published: August 2009

Sterols are constituents of the cellular membranes that are essential for their normal structure and function. In mammalian cells, cholesterol is the main sterol found in the various membranes. However, other sterols predominate in eukaryotic microorganisms such as fungi and protozoa. It is now well established that an important metabolic pathway in fungi and in members of the Trypanosomatidae family is one that produces a special class of sterols, including ergosterol, and other 24-methyl sterols, which are required for parasitic growth and viability, but are absent from mammalian host cells. Currently, there are several drugs that interfere with sterol biosynthesis (SB) that are in use to treat diseases such as high cholesterol in humans and fungal infections. In this review, we analyze the effects of drugs such as (a) statins, which act on the mevalonate pathway by inhibiting HMG-CoA reductase, (b) bisphosphonates, which interfere with the isoprenoid pathway in the step catalyzed by farnesyl diphosphate synthase, (c) zaragozic acids and quinuclidines, inhibitors of squalene synthase (SQS), which catalyzes the first committed step in sterol biosynthesis, (d) allylamines, inhibitors of squalene epoxidase, (e) azoles, which inhibit C14α-demethylase, and (f) azasterols, which inhibitΔ24(25)-sterol methyltransferase (SMT). Inhibition of this last step appears to have high selectivity for fungi and trypanosomatids, since this enzyme is not found in mammalian cells. We review here the IC50 values of these various inhibitors, their effects on the growth of trypanosomatids (both in axenic cultures and in cell cultures), and their effects on protozoan structural organization (as evaluted by light and electron microscopy) and lipid composition. The results show that the mitochondrial membrane as well as the membrane lining the protozoan cell body and flagellum are the main targets. Probably as a consequence of these primary effects, other important changes take place in the organization of the kinetoplast DNA network and on the protozoan cell cycle. In addition, apoptosis-like and autophagic processes induced by several of the inhibitors tested led to parasite death.
hCLCA2 Is a p53-Inducible Inhibitor of Breast Cancer Cell Proliferation
Authors: Walia Vijay , Ding Ming , Kumar Sumit , Nie Daotai , Premkumar Louis S. , Elble Randolph C.
Published: August 2009

hCLCA2 is frequently down-regulated in breast cancer and is a candidate tumor suppressor gene. We show here that the hCLCA2 gene is strongly induced by p53 in response to DNA damage. Adenoviral expression of p53 induces hCLCA2 in a variety of breast cell lines. Further, we find that p53 binds to consensus elements in the hCLCA2 promoter and mutation of these sites abolishes p53-responsiveness and induction by DNA damage. Adenoviral transduction of hCLCA2 into immortalized cells induces p53, CDK inhibitors p21 and p27, and cell cycle arrest by 24 hours, and caspase induction and apoptosis by 40 hours postinfection. Transduction of the malignant tumor cell line BT549 on the other hand does not induce p53, p21, or p27 but instead induces apoptosis directly and more rapidly. Knockout and knockdown studies indicate that growth inhibition and apoptosis are signaled via multiple pathways. Conversely, suppression of hCLCA2 by RNA interference enhances proliferation of MCF10A and reduces sensitivity to doxorubicin. Gene expression profiles indicate that hCLCA2 levels are strongly predictive of tumor cell sensitivity to doxorubicin and other chemotherapeutics. Because certain Cl− channels are proposed to promote apoptosis by reducing intracellular pH, we tested whether, and established that, hCLCA2 enhances Cl− current in breast cancer cells and reduces pH to ∼6.7. These results reveal hCLCA2 as a novel p53-inducible growth inhibitor, explain how its down-regulation confers a survival advantage to tumor cells, and suggest both prognostic and therapeutic applications. [Cancer Res 2009;69(16):6624–32]
Moderate Increase in Mdr1a/1b Expression Causes In vivo Resistance to Doxorubicin in a Mouse Model for Hereditary Breast Cancer
Authors: Pajic Marina , Iyer Jayasree K. , Kersbergen Ariena , van der Burg Eline , Nygren Anders O.H. , Jonkers Jos , Borst Piet , Rottenberg Sven
Published: August 2009

We have found previously that acquired doxorubicin resistance in a genetically engineered mouse model for BRCA1-related breast cancer was associated with increased expression of the mouse multidrug resistance (Mdr1) genes, which encode the drug efflux transporter ATP-binding cassette B1/P-glycoprotein (P-gp). Here, we show that even moderate increases of Mdr1 expression (as low as 5-fold) are sufficient to cause doxorubicin resistance. These moderately elevated tumor P-gp levels are below those found in some normal tissues, such as the gut. The resistant phenotype could be completely reversed by the third-generation P-gp inhibitor tariquidar, which provides a useful strategy to circumvent this type of acquired doxorubicin resistance. The presence of MDR1A in drug-resistant tumors with a moderate increase in Mdr1a transcripts could be shown with a newly generated chicken antibody against a mouse P-gp peptide. Our data show the usefulness of realistic preclinical models to characterize levels of Mdr1 gene expression that are sufficient to cause resistance. [Cancer Res 2009;69(16):6396–9]
Sin3B Expression Is Required for Cellular Senescence and Is Up-regulated upon Oncogenic Stress
Authors: Grandinetti Kathryn B. , Jelinic Petar , DiMauro Teresa , Pellegrino Jessica , Fernández Rodríguez Rubén , Finnerty Patricia M. , Ruoff Rachel , Bardeesy Nabeel , Logan Susan K. , David Gregory
Published: August 2009

Serial passage of primary mammalian cells or strong mitogenic signals induce a permanent exit from the cell cycle called senescence. A characteristic of senescent cells is the heterochromatinization of loci encoding pro-proliferative genes, leading to their transcriptional silencing. Senescence is thought to represent a defense mechanism against uncontrolled proliferation and cancer. Consequently, genetic alterations that allow senescence bypass are associated with susceptibility to oncogenic transformation. We show that fibroblasts genetically inactivated for the chromatin-associated Sin3B protein are refractory to replicative and oncogene-induced senescence. Conversely, overexpression of Sin3B triggers senescence and the formation of senescence-associated heterochromatic foci. Although Sin3B is strongly up-regulated upon oncogenic stress, decrease in expression of Sin3B is associated with tumor progression in vivo, suggesting that expression of Sin3B may represent a barrier against transformation. Together, these results underscore the contribution of senescence in tumor suppression and suggest that expression of chromatin modifiers is modulated at specific stages of cellular transformation. Consequently, these findings suggest that modulation of Sin3B-associated activities may represent new therapeutic opportunities for treatment of cancers. [Cancer Res 2009;69(16):6430–7]
β3 Integrin Subunit Mediates the Bone-Resorbing Function Exerted by Cultured Myeloma Plasma Cells
Authors: Tucci Marco , De Palma Raffaele , Lombardi Lucia , Rodolico Gabriella , Berrino Liberato , Dammacco Franco , Silvestris Franco
Published: August 2009

αvβ3 integrin was investigated in multiple myeloma in relation to the in vitro osteoclast-like activity of malignant plasma cells. Myeloma cells from patients with skeleton involvement overexpressed αvβ3 and produced erosion pits on bone substrates, whereas this effect was not observed by cells from patients with no evidence of bone disease. We therefore explored the αvβ3 transcriptional pathway in the bone-resorbing cells. Silencing of β3 chain abrogated the ability to produce erosion pits and extracellular signal-regulated kinase 1/2 phosphorylation resulting in the defective function of cFos and nuclear factor activator T cell 1, the terminal effectors of osteoclast activation. A similar defect occurred in constitutively β3-deficient cells from patients with no skeleton disease. Microarray gene analysis of β3+ myeloma cells showed that several osteoclast-related genes were up-regulated. Their functions include the activation of receptor pathways β3 and c-fms that regulate several osteoclast functions. These data emphasize the postulated role of myeloma cells in multiple myeloma bone disease and suggest that their osteoclast-like activity is regulated, at least in vitro, by the β3 subunit of the integrin. [Cancer Res 2009;69(16):6738–46]
Small-Molecule Multidrug Resistance–Associated Protein 1 Inhibitor Reversan Increases the Therapeutic Index of Chemotherapy in Mouse Models of Neuroblastoma
Authors: Burkhart Catherine A. , Watt Fujiko , Murray Jayne , Pajic Marina , Prokvolit Anatoly , Xue Chengyuan , Flemming Claudia , Smith Janice , Purmal Andrei , Isachenko Nadezhda , Komarov Pavel G. , Gurova Katerina V. , Sartorelli Alan C. , Marshall Glenn M. , Norris Murray D. , Gudkov Andrei V. , Haber Michelle
Published: August 2009

The multidrug resistance–associated protein 1 (MRP1) has been closely linked to poor treatment response in several cancers, most notably neuroblastoma. Homozygous deletion of the MRP1 gene in primary murine neuroblastoma tumors resulted in increased sensitivity to MRP1 substrate drugs (vincristine, etoposide, and doxorubicin) compared with tumors containing both copies of wild-type MRP1, indicating that MRP1 plays a significant role in the drug resistance in this tumor type and defining this multidrug transporter as a target for pharmacologic suppression. A cell-based readout system was created to functionally determine intracellular accumulation of MRP1 substrates using a p53-responsive reporter as an indicator of drug-induced DNA damage. Screening of small-molecule libraries in this readout system revealed pyrazolopyrimidines as a prominent structural class of potent MRP1 inhibitors. Reversan, the lead compound of this class, increased the efficacy of both vincristine and etoposide in murine models of neuroblastoma (syngeneic and human xenografts). As opposed to the majority of inhibitors of multidrug transporters, Reversan was not toxic by itself nor did it increase the toxicity of chemotherapeutic drug exposure in mice. Therefore, Reversan represents a new class of nontoxic MRP1 inhibitor, which may be clinically useful for the treatment of neuroblastoma and other MRP1-overexpressing drug-refractory tumors by increasing their sensitivity to conventional chemotherapy. [Cancer Res 2009;69(16):6573–80]
Cancer-Specific High-Throughput Annotation of Somatic Mutations: Computational Prediction of Driver Missense Mutations
Authors: Carter Hannah , Chen Sining , Isik Leyla , Tyekucheva Svitlana , Velculescu Victor E. , Kinzler Kenneth W. , Vogelstein Bert , Karchin Rachel
Published: August 2009

Large-scale sequencing of cancer genomes has uncovered thousands of DNA alterations, but the functional relevance of the majority of these mutations to tumorigenesis is unknown. We have developed a computational method, called Cancer-specific High-throughput Annotation of Somatic Mutations (CHASM), to identify and prioritize those missense mutations most likely to generate functional changes that enhance tumor cell proliferation. The method has high sensitivity and specificity when discriminating between known driver missense mutations and randomly generated missense mutations (area under receiver operating characteristic curve, >0.91; area under Precision-Recall curve, >0.79). CHASM substantially outperformed previously described missense mutation function prediction methods at discriminating known oncogenic mutations in P53 and the tyrosine kinase epidermal growth factor receptor. We applied the method to 607 missense mutations found in a recent glioblastoma multiforme sequencing study. Based on a model that assumed the glioblastoma multiforme mutations are a mixture of drivers and passengers, we estimate that 8% of these mutations are drivers, causally contributing to tumorigenesis. [Cancer Res 2009;69(16):OF6660–8]
Proline Oxidase Functions as a Mitochondrial Tumor Suppressor in Human Cancers
Authors: Liu Yongmin , Borchert Gregory L. , Donald Steven P. , Diwan Bhalchandra A. , Anver Miriam , Phang James M.
Published: August 2009

Tumor metabolism and bioenergetics have become important topics for cancer research and are promising targets for anticancer therapy. Although glucose serves as the main source of energy, proline, an alternative substrate, is important, especially during nutrient stress. Proline oxidase (POX), catalyzing the first step in proline catabolism, is induced by p53 and can regulate cell survival as well as mediate programmed cell death. In a mouse xenograft tumor model, we found that POX greatly reduced tumor formation by causing G2 cell cycle arrest. Furthermore, immunohistochemical staining showed decreased POX expression in tumor tissues. Importantly, HIF-1α signaling was impaired with POX expression due to the increased production of α-ketoglutarate, a critical substrate for prolyl hydroxylation and degradation of HIF-1α. Combined with previous in vitro findings and reported clinical genetic associations, these new findings lead us to propose POX as a mitochondrial tumor suppressor and a potential target for cancer therapy. [Cancer Res 2009;69(16):6414–22]
Biomarkers of Dietary Energy Restriction in Women at Increased Risk of Breast Cancer
Authors: Ong Kai Ren , Sims Andrew H. , Harvie Michelle , Chapman Mary , Dunn Warwick B. , Broadhurst David , Goodacre Royston , Wilson Mary , Thomas Nicola , Clarke Robert B. , Howell Anthony
Published: August 2009

Dietary energy restriction (DER) reduces risk of spontaneous mammary cancer in rodents. In humans, DER in premenopausal years seems to reduce risk of postmenopausal breast cancer. Markers of DER are required to develop acceptable DER regimens for breast cancer prevention. We therefore examined markers of DER in the breast, adipose tissue, and serum.

Nineteen overweight or obese women at moderately increased risk of breast cancer (lifetime risk, 1 in 6 to 1 in 3) ages between 35 and 45 were randomly allocated to DER [liquid diet, 3,656 kJ/d (864 kcal/d); n = 10] or asked to continue their normal eating patterns (n = 9) for one menstrual cycle. Biopsies of the breast and abdominal fat were taken before and after the intervention. RNA was extracted from whole tissues and breast epithelium (by laser capture microdissection) and hybridized to Affymetrix GeneChips. Longitudinal plasma and urine samples were collected before and after intervention, and metabolic profiles were generated using gas chromatography-mass spectrometry.

DER was associated with significant reductions in weight [−7.0 (±2.3) kg] and in alterations of serum biomarkers of breast cancer risk (insulin, leptin, total and low-density lipoprotein cholesterol, and triglycerides). In both abdominal and breast tissues, as well as isolated breast epithelial cells, genes involved in glycolytic and lipid synthesis pathways (including stearoyl-CoA desaturase, fatty acid desaturase, and aldolase C) were significantly down-regulated.

We conclude that reduced expressions of genes in the lipid metabolism and glycolytic pathways are detectable in breast tissue following DER, and these may represent targets for DER mimetics as effective chemoprophylactic agents.
Expression of Cochlin mRNA Splice Variants in the Inner Ear
Authors: Sekine Kuwon , Ikezono Tetsuo , Matsumura Tomohiro , Shindo Susumu , Watanabe Atsushi , Li Lishu , Pawankar Ruby , Nishino Takeshi , Yagi Toshiaki
Published: August 2009

Proteomic analysis of inner ear proteins revealed unique properties of cochlin, encoded by the COCH gene. We detected 3 cochlin isoforms, p63s, p44s and p40s, in the inner ear tissue and a short 16-kDa isoform, cochlin-tomoprotein (CTP), in the perilymph. The role of the cochlin isoforms has not been elucidated. To improve our understanding of the mechanism of cochlin isoform expression, we investigated rat cochlin mRNA expression in the inner ear and other organs. We performed RNA-ligation-mediated amplification of cDNA ends (RLM-RACE) using RNA isolated from the inner ear and spleen of rats, which are known to express abundant cochlin mRNA. We also examined the expression profile of full-length cochlin mRNA by nested RT-PCR in the cerebrum, cerebellum/brain stem, eye, inner ear, thyroid gland, thymus gland, lung, heart, liver, spleen, adrenal gland, kidney and blood. We verified CTP expression in rat perilymph by Western blot. By RLM-RACE, alternately spliced variants of cochlin mRNA with 3 different lengths were detected (2442, 2008 and 724 bp). The two longer mRNAs encode full-length cochlin with different polyadenylation signals in the 3′-untranslated region, which are expressed both in the ear and spleen. The short variant encodes the limulus factor C, cochlin, late gestation lung protein (LCCL) domain and the N-terminal sequence of the von Willebrand factor A (vWFA1) domain, and this variant was detected only in the ear. All 3 variants have the same transcriptional start site. By RT-PCR, we found that full-length cochlin was expressed in all organs examined, with a splice variant in the heart. By Western blot, we detected short isoforms (11–17 kDa) in the perilymph. Cochlin isoform formation is regulated, at least in part, by alternative splicing at the transcriptional level. The short mRNA was detected only in the inner ear, and this variant may provide a clue to understanding the formation and function of cochlin isoforms.
Tuberculin Anergy in Clinically Normal Individuals
Authors: Muller Konrad , Pye Daryl W. , Martin Leon , Kimpton Wayne G.
Published: August 2009

Tuberculin anergy was demonstrated in a number of clinically normal individuals who, on being immunized with Bacillus Calmette-Guérin (BCG), failed to demonstrate a positive Mantoux skin test. Lymphoid cells from these anergic subjects when stimulated with purified protein derivative (PPD) in vitro did not produce macrophage migration inhibition factor (MIF). Lymphocyte transformation studies demonstrated that while Mantoux-positive, non-BCG immunized Mantoux-negative and anergic individuals could all undergo blast cell transformation in the presence of PHA, only lymphoid cells from Mantoux-positive and a proportion of anergic people were capable of blast cell transformation when cultured with PPD. These results suggest that more than one mechanism may account for tuberculin anergy in clinically normal people operating at the level of lymphokine production and in some cases lymphocyte transformation.
The Mechanism of Passive Sensitization: Occupation of Free IgE Receptors or Exchange with Cell-Bound IgE
Authors: Van Toorenenbergen A.W. , Aalberse R.C , Reerink-Brongers Eveline E.
Published: August 2009

Leukocytes were passively sensitized, as judged by allergen-induced histamine release. Before and after passive sensitization, the amount of total IgE on basophil leukocytes was measured by quantitative immunofluorescence microscopy. No measurable increase in IgE load was observed. When leukocytes were incubated with acid buffer after passive sensitization, no allergen-specific IgE was found in acid eluates of these leukocytes. Preincubation of leukocytes with excess irrelevant IgE resulted in inhibition of a subsequent passive sensitization. Postincubation of in vivo- or in vitro-sensitized leukocytes with excess irrelevant IgE had no effect on the sensitivity of these cells towards allergen. When leukocytes were incubated with TNP-labeled myeloma IgE and subsequently with fluorescein-labeled anti-TNP antibodies, no fluorescence was observed on basophil leukocytes, although binding of TNP-labeled IgE was demonstrated by anti-TNP antiserum-induced histamine release. It is concluded that only small amounts of IgE become bound to basophil leukocytes during passive sensitization, compared with the amounts of IgE already present on these cells. Exchange of IgE between cells and sensitizing serum does not take place to a measurable extent during passive sensitization.
Passive Anaphylaxis in Human Lung Fragments as a Model for Testing Anti-Allergic Drugs: Its Variability and Constraints
Authors: Young Kevin D. , Church Martin K.
Published: August 2009

Histamine release from human lung fragments, passively sensitized and challenged with antigen under standardized experimental conditions, varied between 0 and 41.2% (mean ± SD 15.6 ± 10.0%) in 89 experiments. Over 41 lungs, the mean coefficient of variation for release from 10 to 28 tissue replicates was 21.4% (range 7–44%). Sodium cromoglycate and chlorpromazine were both partial antagonists of histamine release producing, at best, 30–40% inhibition. The cromoglycate analogue, bufrolin, showed similar activity. There was considerable variation in the effects of these drugs both within and between experiments. Salbutamol was a more potent and more effective inhibitor of release but it, too, showed variability. Although theoretically a very appropriate model of allergic asthma, passive anaphylaxis in human lung fragments is quantitatively inconsistent and gives only a gross indication of drug efficacy.
Cow’s Milk Protein Intolerance in Children is Connected with Impaired Lymphoblastic Responses to Mitogens
Authors: Fällström S.-P. , Lindholm L. , Ahlstedt S.
Published: August 2009

The lymphoblastic responses of peripheral blood lymphocytes to mitogens were analyzed in 36 infants and children aged 1–28 months. The patients had confirmed or suspected cow’s milk protein intolerance (CMPI). Immediately after a positive provocation test 12 out of 14 patients with repeatedly confirmed CMPI showed more than 30% decrease of their lymphoblastic responses to pokeweed mitogen and cancanavalin A compared with their initial values. In 10 infants the response to phytohemagglutinin was similarly decreased. In 4 out of 12 infants with previously confirmed CMPI, but with no reaction at challenge, similar decreases were seen to some of the mitogens. In patients with suspected CMPI, but with no obvious reaction to the challenge with cow’s milk, generally an increase of the lymphoblastic responses was seen compared with the inital values.
Granulocyte-Mediated Release of Histamine from Mast Cells
Authors: Stendahl O. , Molin L. , Lindroth M.
Published: August 2009

During phagocytosis, neutrophilic and eosinophilic granulocytes undergo metabolic activation and degranulation. This leads to accumulation of H2O2 and myeloperoxidase (MPO) or eosinophilic peroxidase (EPO) in the extracellular environment in inflammation. In the present investigation we show that the MPO-H2O2-halide system can degranulate isolated mast cells with release of histamine and 3H-serotonin. This is accomplished both in a system containing purified human MPO and in one containing phagocytosing neutrophils. When using purified MPO and a H2O2-generating system, the release was rapid, with 50% release after less than 20 min. The direct histamine release required less (0.65–2 μg/ml) MPO than did the release of 3H-serotonin (1–6 μg/ml). The release reaction was inhibited by azide, catalase but not superoxide dismutase. Studying the effect of the antiinflammatory sulfone compounds dapsone and sulfapyridine on the MPO-mediated histamine release, we show effective inhibition of the histamine release at concentrations of 0.025–0.2 mM of sulfone. 5-aminosalicylic acid exhibited some inhibition only in the cell-free system, whereas sulfasalazine was inactive. The antiinflammatory effect of dapsone in dermatitis herpetiformis and sulfasalazine in ulcerative colitis may in part be explained by their inhibitory effects on the granulocyte-mediated histamine release from mast cells in skin and mucosal tissue.
Properties of Two Monoclonal Antibodies Directed against the Fc and Fab′ Regions of Rat IgE
Authors: Conrad Daniel H. , Studer Elaine , Gervasoni Jim , Mohanakumar T.
Published: August 2009

Hybridoma antibodes directed against the Fc and Fab portions of rat IgE were produced by immunizing BALB/c mice with rat IgE and fusing the spleen cells with the nonsecreting plasmacytoma P3/X63Ag8.653. Two of the antibodies, designated as A2 and B5, were extensively characterized. Competitive binding experiments using rat IgE from the IR 162 and IR 2 immunocytomas and rat IgG indicated that both A2 and B5 were ε-chain specific and not anti-idiotype. A2 also exhibited some cross-reactivity with mouse IgE. When IgE was treated with chymotrypsin so as to produce both F(ab′)2 and Fab fragments, the enzyme-treated IgE retained reactivity with B5, but the reactivity to A2 was lost. Heat denaturation of IgE at 56 °C resulted in a progressive loss of reactivity of the IgE for both A2 and the Fc receptor on rat basophilic leukemia cells; the reactivity of B5 remained unchanged. A2 does not evidently interact with the same site on the Fc of IgE that is involved in binding to the rat basophilic leukemia cell Fc receptor; A2 exerted little influence on the binding of IgE to rat basophilic leukemia cells. Thus, the data indicate that the antigenic site for B5 is in the Fab region of the IgE molecule and that A2 reacts with the IgE Fc. Use of these antibodies to measure cell-bound IgE was also evaluated in dual label experiments, and potential problems in using divalent antibodies to quantitate cell surface antigens are discussed.
A Simple Solid Phase Radioimmunoassay Specific for Human C3b to Detect C3b Receptors on Human Lymphoblastoid Cell Surfaces
Authors: Barel Monique , Charriaut Christiane , Frade Raymond
Published: August 2009

Our aim was to detect C3b receptors on human lymphoblastoid cells using a solid phase radioimmunoassay (RIA) specific for human C3b. RIA was performed by coupling rabbit antihuman C3b to acrylamide beads to make immunobeads. The specificity and sensitivity of binding of 125I-C3b to immunobeads allowed the detection of as little as 6 × 10––10M unlabeled human C3b. The cells were incubated with a C3b concentration (10––9M) giving 25 % inhibition in the RIA. The concentration of unbound C3b was then measured in the cell supernatants using RIA. Results showed that: (a) loss of C3b antigen in the cell supernatant was not due to degradation of C3b molecules, (b) C3b binding could be detected at 37 °C on the four B cell lines, but not on the two T cell lines or on the two non T-non B cell lines tested, (c) C3b bound on the B lymphoblastoid cells was not cleaved, neither into iC3b nor C3c and C3d fragments, supporting the presence of C3b receptors on the cells tested. This method allows screening of a variety of C3b receptor-positive cells.
Enhancement of in vitro and in vivo Antitumor Activity by Cord Factor (6-6′-Dimycolate of Trehalose) Administered Suspended in Saline
Authors: Orbach-Arbouys Simone , Tenu J.P. , Petit J.F.
Published: August 2009

The antitumor activity of trehalose-6,6′-dimycolate (TDM) dispersed in saline was studied in vitro and in vivo. In vitro, macrophages from TDM-treated mice entirely abolish, even at a 1.25:1 effector/target cell ratio, the (3H)TdR incorporation of P815 mastocytoma cells to the culture in which they were added. In vivo, the number of P815 cells harvested 10 days after their injection into TDM-treated mice was reduced by a factor higher than 50. The slopes of the radiolabel elimination of TDM-treated mice injected with 107125I-UdR-labelled L1210 leukemic cells were greater than those of untreated controls. The interest of such a compound which seems to be one of the active structures responsible for the antitumor activity of BCG is its efficiency when administered dispersed in saline.
Cyclic AMP Levels during Stimulation and Inhibition of Histamine Release from Rat Mast Cells by the Calmodulin Inhibitor Trifluoperazine
Authors: Alm Per E.
Published: August 2009

The calmodulin inhibitor trifluoperazine represses compound 48/80-induced histamine release. The effects appear to take place at a step distal to the entrance of calcium into the cell. Trifluoperazine in itself induces histamine release from rat peritoneal mast cells. This release is totally independent of extracellular calcium and at high concentrations of the drug (≥20 μM), it is due to lysis of the cells. Trifluoperazine reduces the compound 48/80-induced decrease in mast cell cyclic AMP content, probably by inhibiting a cyclic nucleotide phosphodiesterase. The trifluoperazine-induced repression of histamine release does not seem to be correlated to the ability of the drug to alter the compound 48/80 induced changes in cyclic AMP.
Studies in Immunodermatology
Authors: Kumar Vijay , Rogozinski Tomasz , Beutner Ernst H. , Jablonska Stefania
Published: August 2009

Explants of normal skin fail to react in direct immunofluorescence tests with stratum corneum antibodies. However, upon stripping with cellophane tape, the horny layer of such explants react in tissue culture. Swabbing of skin explants with ether and chloroform converts stratum corneum antigen (SCAg) from a non-reactive to a reactive form. Treatment with methanol, acetone or phosphate-buffered saline failed to bring about such a conversion. Treatment of skin explants with hyaluronidase and phosphilipase A converst SCAg of at least some skin explants to a reactive form. Treatment with trypsin, chymotrypsin and plasmin abolished the reactivity of SCAg upon prolonged incubation. However, upon short incubation with plasmin, SCAg was converted to a reactive form.
Half-Life of Murine IgE Antibodies in the Mouse
Authors: Hirano Takao , Hom Christine , Ovary Zoltan
Published: August 2009

The half-life of intravenously injected murine anti-DNP IgE antibody was determined in recipient mice by sequential bleeding and was found to be 12 h. Intradermally injected IgE antibody persisted much longer, the half-life was 6 days.