bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro processing and secretion of mutant insulin proteins that cause permanent neonatal diabetes
Authors: Rajan Sindhu , Eames Stefani C. , Park Soo-Young , Labno Christine , Bell Graeme I. , Prince Victoria E. , Philipson Louis H.
Published: December 2009

Permanent neonatal diabetes mellitus is a rare form of insulin-requiring diabetes presenting within the first few weeks or months of life. Mutations in the insulin gene are the second most common cause of this form of diabetes. These mutations are located in critical regions of preproinsulin and are likely to prevent normal processing or folding of the preproinsulin/proinsulin molecule. To characterize these mutations, we transiently expressed proinsulin-GFP fusion proteins in MIN6 mouse insulinoma cells. Our study revealed three groups of mutant proteins: 1) mutations that result in retention of proinsulin in the endoplasmic reticulum (ER) and attenuation of secretion of cotransfected wild-type insulin: C43G, F48C, and C96Y; 2) mutations with partial ER retention, partial recruitment to granules, and attenuation of secretion of wild-type insulin: G32R, G32S, G47V, G90C, and Y108C; and 3) similar to ( 2) but with no significant attenuation of wild-type insulin secretion: A24D and R89C. The mutant insulin proteins do not prevent targeting of wild-type insulin to secretory granules, but most appear to lead to decreased secretion of wild-type insulin. Each of the mutants triggers the expression of the proapoptotic gene Chop, indicating the presence of ER stress.
Binge-pattern alcohol exposure during puberty induces sexually dimorphic changes in genes regulating the HPA axis
Authors: Przybycien-Szymanska Magdalena M. , Rao Yathindar S. , Pak Toni R.
Published: December 2009

Maternal alcohol consumption during critical periods of fetal brain development leads to devastating long-term consequences on adult reproductive physiology, cognitive function, and social behaviors. However, very little is known about the long-term consequences of alcohol consumption during puberty, which is perhaps an equally dynamic and critical period of brain development. Alcohol abuse during adulthood has been linked with an increase in clinically diagnosed anxiety disorders, yet the etiology and neurochemical mechanisms of alcohol-induced anxiety behavior is unknown. In this study, we determined the effects of binge ethanol exposure during puberty on two critical central regulators of stress and anxiety behavior: corticotrophin-releasing hormone (CRH) and arginine vasopressin (AVP). Our results showed that ethanol increased plasma corticosterone (CORT) levels in both sexes, yet binge-treated animals had significantly lower CORT levels than animals exposed to a single dose, suggesting that the hypothalamo-pituitary-adrenal (HPA) axis habituated to the repeated stressful stimuli of ethanol. Binge ethanol exposure also significantly increased CRH and AVP gene expression in the paraventricular nucleus of males, but not females. Overall, our results demonstrate that binge ethanol exposure during puberty changes the central expression of stress-related genes in a sex-specific manner, potentially leading to permanent dysregulation of the HPA axis and long-term behavioral consequences.
Deep Brain Stimulation Does Not Silence Neurons in Subthalamic Nucleus in Parkinson's Patients
Authors: Carlson Jonathan D. , Cleary Daniel R. , Cetas Justin S. , Heinricher Mary M. , Burchiel Kim J.
Published: December 2009

Two broad hypotheses have been advanced to explain the clinical efficacy of deep brain stimulation (DBS) in the subthalamic nucleus (STN) for treatment of Parkinson's disease. One is that stimulation inactivates STN neurons, producing a functional lesion. The other is that electrical stimulation activates the STN output, thus “jamming” pathological activity in basal ganglia-corticothalamic circuits. Evidence consistent with both concepts has been adduced from modeling and animal studies, as well as from recordings in patients. However, the stimulation parameters used in many recording studies have not been well matched to those used clinically. In this study, we recorded STN activity in patients with Parkinson's disease during stimulation delivered through a clinical DBS electrode using standard therapeutic stimulus parameters. A microelectrode was used to record the firing of a single STN neuron during DBS (3–5 V, 80–200 Hz, 90- to 200-μs pulses; 33 neurons/11 patients). Firing rate was unchanged during the stimulus trains, and the recorded neurons did not show prolonged (s) changes in firing rate on termination of the stimulation. However, a brief (∼1 ms), short-latency (6 ms) postpulse inhibition was seen in 10 of 14 neurons analyzed. A subset of neurons displayed altered firing patterns, with a predominant shift toward random firing. These data do not support the idea that DBS inactivates the STN and are instead more consistent with the hypothesis that this stimulation provides a null signal to basal ganglia-corticothalamic circuitry that has been altered as part of Parkinson's disease.
Responses of Ventral Posterior Thalamus Neurons to Three-Dimensional Vestibular and Optic Flow Stimulation
Authors: Meng Hui , Angelaki Dora E.
Published: December 2009

Multisensory neurons tuned to both vestibular and visual motion (optic flow) signals are found in several cortical areas in the dorsal visual stream. Here we examine whether such convergence occurs subcortically in the macaque thalamus. We searched the ventral posterior nuclei, including the anterior pulvinar, as well as the ventro-lateral and ventral posterior lateral nuclei, areas that receive vestibular signals from brain stem and deep cerebellar nuclei. Approximately a quarter of cells responded to three-dimensional (3D) translational and/or rotational motion. More than half of the responsive cells were convergent, thus responded during both rotation and translation. The preferred axes of translation/rotation were distributed throughout 3D space. The majority of the neurons were excited, but some were inhibited, during rotation/translation in darkness. Only a couple of neurons were multisensory being tuned to both vestibular and optic flow stimuli. We conclude that multisensory vestibular/optic flow neurons, which are commonly found in cortical visual and visuomotor areas, are rare in the ventral posterior thalamus.
Fish nutrients decrease expression levels of tumor necrosis factor-α in cultured human macrophages
Authors: Rudkowska Iwona , Marcotte Bruno , Pilon Geneviève , Lavigne Charles , Marette André , Vohl Marie-Claude
Published: December 2009

Numerous studies have demonstrated the beneficial effects of fish consumption on inflammatory markers. Until now, these beneficial effects of fish consumption have been mostly linked to the omega-3 fatty acids (FA). The objective of the present study was to examine, in vitro, whether expression levels of genes involved in the inflammatory response differ in human macrophages incubated with casein hydrolysates (CH) or fish protein hydrolysates (FPH) in the presence or absence of omega-3 FA compared with omega-3 FA alone. Peripheral blood monocytes differentiated into macrophages from 10 men were incubated in the presence of omega-3 FA (10 μM eicosapentaenoic acid and 5 μM docosahexaenoic acid) or CH or FPH (10, 100, 1,000 μg) with or without omega-3 FA for 48 h. Results demonstrate that expression levels of tumor necrosis factorα ( TNFα) had a tendency to be lower after the addition of FPH alone or CH with omega-3 FA compared with omega-3 FA treatment. Furthermore, the combination of FPH and omega-3 FA synergistically decreased expression levels of TNFα compared to treatment with omega-3 FA or FPH alone. No difference on gene expression levels of interleukin-6 was observed between treatments. In conclusion, these preliminary results suggest that the anti-inflammatory effects of fish consumption can be explained by a synergistic effect of the omega-3 FA with the protein components of fish on TNFα expression and therefore contribute to the beneficial effects of fish consumption. Hence, follow-up studies should be performed to confirm the effects of a diet rich in FPH and omega-3 FA on serum proinflammatory cytokine concentrations.
Genes controlling postural changes in blood pressure: comprehensive association analysis of ATP-sensitive potassium channel genes KCNJ8 and ABCC9
Authors: Ellis Justine A. , Lamantia Angela , Chavez Raul , Scurrah Katrina J. , Nichols Colin G. , Harrap Stephen B.
Published: December 2009

Buffering of blood pressure during change of posture such as standing is controlled largely by the baroreflex. In our population-based Victorian Family Heart Study (VFHS), we previously demonstrated that, on average, systolic blood pressure (SBP) changes very little on standing; however, interindividual variation is substantial and shows familial aggregation, with ∼25% of the variance attributable to genetic factors. Our genomewide linkage analysis suggests a region on chromosome 12p that harbors two strong candidate genes, KCNJ8 and ABCC9, encoding the channel-forming inward rectifier subunit Kir6.1 and the ATP-sensitive binding cassette SUR2B, respectively. These are key components of smooth muscle ATP-sensitive potassium (KATP) channels, important regulators of arterial tone and blood flow and central to autonomic baroreceptor control of changes in total peripheral resistance. We performed a comprehensive association analysis of 47 tag single nucleotide polymorphisms (SNPs) spanning the KCNJ8 and ABCC9 gene regions with postural change in SBP (ΔSBP). To augment power, we took a selective genotyping approach in which we compared allele and genotype frequencies between 150 unrelated individuals with high (positive) ΔSBP (≥ 7 mmHg) and 150 unrelated individuals with low (negative) ΔSBP (≤ −7 mmHg) drawn from the offspring generation (18–30 yr) of the VFHS. Association analyses showed that no SNPs demonstrated statistically significant differences in genotype frequencies between groups, particularly after adjustments for multiple testing. We conclude that sequence variants in KCNJ8 and ABCC9 are unlikely to contribute to variation in ΔSBP. Other genes in the identified chromosome 12p region warrant investigation.
The Potential to Forgo Social Welfare Gains through Overreliance on Cost Effectiveness/Cost Utility Analyses in the Evidence Base for Public Health
Authors: Cohen D. R. , Patel N.
Published: December 2009

Economic evaluations of clinical treatments most commonly take the form of cost effectiveness or cost utility analyses. This is appropriate since the main—sometimes the only—benefit of such interventions is increased health. The majority of economic evaluations in public health, however, have also been assessed using these techniques when arguably cost benefit analyses would in many cases have been more appropriate, given its ability to take account of nonhealth benefits as well. An examination of the nonhealth benefits from a sample of studies featured in a recent review of economic evaluations in public health illustrates how overfocusing on cost effectiveness/cost utility analyses may lead to forgoing potential social welfare gains from programmes in public health. Prior to evaluation, programmes should be considered in terms of the potential importance of nonhealth benefits and where these are considerable would be better evaluated by more inclusive economic evaluation techniques.
A Boundary Value Problem with Multivariables Integral Type Condition for Parabolic Equations
Authors: Marhoune A. L. , Lakhal F.
Published: December 2009

We study a boundary value problem with multivariables integral type condition for a class of parabolic equations. We prove the existence, uniqueness, and continuous dependence of the solution upon the data in the functional wieghted Sobolev spaces. Results are obtained by using a functional analysis method based on two‐sided a priori estimates and on the density of the range of the linear operator generated by the considered problem.
WNT16B Is a New Marker of Cellular Senescence That Regulates p53 Activity and the Phosphoinositide 3-Kinase/AKT Pathway
Authors: Binet Romuald , Ythier Damien , Robles Ana I. , Collado Manuel , Larrieu Delphine , Fonti Claire , Brambilla Elisabeth , Brambilla Christian , Serrano Manuel , Harris Curtis C. , Pedeux Rémy
Published: December 2009

Senescence is a tumor suppression mechanism that is induced by several stimuli, including oncogenic signaling and telomere shortening, and controlled by the p53/p21WAF1 signaling pathway. Recently, a critical role for secreted factors has emerged, suggesting that extracellular signals are necessary for the onset and maintenance of senescence. Conversely, factors secreted by senescent cells may promote tumor growth. By using expression profiling techniques, we searched for secreted factors that were overexpressed in fibroblasts undergoing replicative senescence. We identified WNT16B, a member of the WNT family of secreted proteins. We found that WNT16B is overexpressed in cells undergoing stress-induced premature senescence and oncogene-induced senescence in both MRC5 cell line and the in vivo murine model of K-RasV12–induced senescence. By small interfering RNA experiments, we observed that both p53 and WNT16B are necessary for the onset of replicative senescence. WNT16B expression is required for the full transcriptional activation of p21WAF1. Moreover, WNT16B regulates activation of the phosphoinositide 3-kinase (PI3K)/AKT pathway. Overall, we identified WNT16B as a new marker of senescence that regulates p53 activity and the PI3K/AKT pathway and is necessary for the onset of replicative senescence. [Cancer Res 2009;69(24):9183–91]
Epithelial-to-Mesenchymal Transition and Ovarian Tumor Progression Induced by Tissue Transglutaminase
Authors: Shao Minghai , Cao Liyun , Shen Changyu , Satpathy Minati , Chelladurai Bhadrani , Bigsby Robert M. , Nakshatri Harikrishna , Matei Daniela
Published: December 2009

Tissue transglutaminase (TG2), an enzyme that catalyzes Ca2+-dependent aggregation and polymerization of proteins, is overexpressed in ovarian cancer cells and tumors. We previously reported that TG2 facilitates tumor dissemination using an i.p. xenograft model. Here we show that TG2 modulates epithelial-to-mesenchymal transition (EMT), contributing to increased ovarian cancer cell invasiveness and tumor metastasis. By using stable knockdown and overexpression in epithelial ovarian cancer cells, we show that TG2 induces a mesenchymal phenotype, characterized by cadherin switch and invasive behavior in a Matrigel matrix. This is mediated at the transcriptional level by altering the expression levels and function of several transcriptional repressors, including Zeb1. One mechanism through which TG2 induces Zeb1 is by activating the nuclear factor-κB complex. The effects of TG2 on ovarian cancer cell phenotype and invasiveness translate into increased tumor formation and metastasis in vivo, as assessed by an orthotopic ovarian xenograft model. Highly expressed in ovarian tumors, TG2 promotes EMT and enhances ovarian tumor metastasis by activating oncogenic signaling. [Cancer Res 2009;69(24):9192–201]
Tgfbr1 Haploinsufficiency Inhibits the Development of Murine Mutant Kras-Induced Pancreatic Precancer
Authors: Adrian Kevin , Strouch Matthew J. , Zeng Qinghua , Barron Morgan R. , Cheon Eric C. , Honasoge Akilesh , Xu Yanfei , Phukan Sharbani , Sadim Maureen , Bentrem David J. , Pasche Boris , Grippo Paul J.
Published: December 2009

To dissect the role of constitutively altered Tgfbr1 signaling in pancreatic cancer development, we crossed Elastase-KrasG12D (EL-Kras) mice with Tgfbr1 haploinsufficient mice to generate EL-Kras/Tgfbr1+/− mice. Mice were euthanized at 6 to 9 months to compare the incidence, frequency, and size of precancerous lesions in the pancreas. Only 50% of all EL-Kras/Tgfbr1+/− mice developed preinvasive lesions compared with 100% of EL-Kras (wild-type Tgfbr1) mice. The frequency of precancerous lesions was 4-fold lower in haploinsufficient than in control mice. Paradoxically, the precancerous lesions of EL-Kras/Tgfbr1+/− mice were considerably larger than those in EL-Kras mice. Yet, the mitotic index of precancerous cells and the observable levels of fibrosis, lipoatrophy, and lymphocytic infiltration were reduced in EL-Kras/Tgfbr1+/− mice. We conclude that Tgfbr1 signaling promotes the development of precancerous lesions in mice. These findings suggest that individuals with constitutively decreased TGFBR1 expression may have a decreased risk of pancreatic cancer. [Cancer Res 2009;69(24):9169–74]
Cancer Stem Cells and Aneuploid Populations within Developing Tumors Are the Major Determinants of Tumor Dormancy
Authors: Kusumbe Anjali P. , Bapat Sharmila A.
Published: December 2009

Tumor formation involves substantial cell division and genetic instability, but the relationship between quiescent cancer stem cells (CSC) and dividing progenitors in these events is poorly understood. Likewise, the implication of aneuploid cells in solid tumors is uncertain. CSCs are postulated to contribute to tumor dormancy and present a formidable obstacle in limiting treatment outcomes for a majority of cancers, whereas the genetic heterogeneity conjured by aneuploid cells may influence tumor drug resistance. However, direct confirmation of these events remains forthcoming. In the present study, we addressed the identification of tumor dormancy in terms of isolation of therapy-refractory residual tumor cells from tumors that persist in a state of quiescence as label-retaining cells. The choices of label were PKH67/PKH26 dyes that irreversibly bind to the lipid bilayer on cell membranes and get equally partitioned among daughter cells subsequent to each cell division. Consequent characterization revealed that label-retaining cells encompass two different populations capable of remaining in a state of quiescence, i.e., stem-like cells and aneuploid cells. The former express a reversibility of quiescence through retention of functionality and also exhibit therapeutic refractoriness; the latter seem to be either quiescent or proliferation-arrested at steady-state. Subsequent to exposure to selective pressure of chemotherapy, a fraction of these cells may acquire the potential to proliferate in a drug-refractory manner and acquire stem-like characteristics. Collectively, the findings of the present study reveal that tumor-derived CSCs and aneuploid populations contribute to drug resistance and tumor dormancy in cancer progression. [Cancer Res 2009;69(24):9245–53]
Targeting heat shock protein 90 with CUDC-305 overcomes erlotinib resistance in non–small cell lung cancer
Authors: Bao Rudi , Lai Cheng-Jung , Wang Da-Gong , Qu Hui , Yin Ling , Zifcak Brian , Tao Xu , Wang Jing , Atoyan Ruzanna , Samson Maria , Forrester Jeffrey , Xu Guang-Xin , DellaRocca Steven , Borek Mylissa , Zhai Hai-Xiao , Cai Xiong , Qian Changgeng
Published: December 2009

CUDC-305 is a heat shock protein 90 (HSP90) inhibitor of the novel imidazopyridine class. Here, we report its activities in non–small cell lung cancer (NSCLC) cell lines with gene deregulations conferring primary or secondary resistance to epidermal growth factor receptor (EGFR) inhibitors. We show that CUDC-305 binds strongly to HSP90 extracted from erlotinib-resistant NSCLC cells (IC50 70 nmol/L). This result correlates well with the potent antiproliferative activity in erlotinib-resistant NSCLC cell lines (IC50 120–700 nmol/L) reported previously. Furthermore, it exhibits durable inhibition of multiple oncoproteins and induction of apoptosis in erlotinib-resistant NSCLC cells. CUDC-305 potently inhibits tumor growth in subcutaneous xenograft models of H1975 and A549, which harbor EGFR T790M mutation or K-ras mutations conferring acquired and primary erlotinib resistance, respectively. In addition, CUDC-305 significantly prolongs animal survival in orthotopic lung tumor models of H1975 and A549, which may be partially attributed to its preferential exposure in lung tissue. Furthermore, CUDC-305 is able to extend animal survival in a brain metastatic model of H1975, further confirming its ability to cross the blood-brain barrier. Correlating with its effects in various tumor models, CUDC-305 induces degradation of receptor tyrosine kinases and downstream signaling molecules of the PI3K/AKT and RAF/MEK/ERK pathways simultaneously, with concurrent induction of apoptosis in vivo. In a combination study, CUDC-305 enhanced the antitumor activity of a standard-of-care agent in the H1975 tumor model. These results suggest that CUDC-305 holds promise for the treatment of NSCLC with primary or acquired resistance to EGFR inhibitor therapy. [Mol Cancer Ther 2009;8(12):3296–306]
A novel two-step transcriptional activation system for gene therapy directed toward epithelial cells
Authors: Arendt Maja L. , Nasir Lubna , Morgan Iain M.
Published: December 2009

The two-step transcriptional activation (TSTA) mechanism in gene therapy amplifies cell type–specific promoter activity, allowing for increased levels of gene expression in target tissues. In this system, the specific promoter drives expression of a strong transcriptional activator that binds to DNA target sequences located upstream from a second promoter controlling the expression of the therapeutic gene. The majority of previous studies have exploited a fusion between the DNA binding domain of the yeast transcriptional activator Gal4 fused to the VP16 activation domain of herpes simplex virus 1 as the transcriptional activator. In this report, an alternative to this system is described based on a fusion protein containing the DNA binding domain of the bovine papillomavirus 1 transcriptional activator E2 fused to VP16 that induces target gene expression following binding to a minimal bovine papillomavirus 4 promoter containing upstream E2 binding sites and only 3 bp of promoter sequence upstream from the TATA box. VP16-E2 is superior to Gal4-VP16 as the transcriptional activator in a TSTA system driven by either of the two potentially cancer-specific promoters telomerase RNA and telomerase reverse transcriptase in several cell lines. Results also suggest that this new system has an advantage in epithelial cells and is therefore ideal for potential targeting of carcinomas. By incorporating the TRAIL gene as a transgene in the VP16-E2 TSTA system, selective killing of telomerase-positive cells occurs. We propose that our new system should be considered in future TSTA, particularly when targeting epithelial-derived cells. [Mol Cancer Ther 2009;8(12):3244–54]
The antiproliferative activity of the heat shock protein 90 inhibitor IPI-504 is not dependent on NAD(P)H:quinone oxidoreductase 1 activityin vivo
Authors: Douglas Mark , Lim Alice R. , Porter James R. , West Kip , Pink Melissa M. , Ge Jie , Wylie Andrew A. , Tibbits Thomas T. , Biggs Kurtis , Curtis Michael , Palombella Vito J. , Adams Julian , Fritz Christian C. , Normant Emmanuel
Published: December 2009

IPI-504, a water-soluble ansamycin analogue currently being investigated in clinical trials, is a potent inhibitor of the protein chaperone heat shock protein 90 (Hsp90). Inhibition of Hsp90 by IPI-504 triggers the degradation of important oncogenic client proteins. In cells, the free base of IPI-504 hydroquinone exists in a dynamic redox equilibrium with its corresponding quinone (17-AAG); the hydroquinone form binding 50 times more tightly to Hsp90. It has been proposed recently that the NAD(P)H:quinone oxidoreductase NQO1 can produce the active hydroquinone and could be essential for the activity of IPI-504. Here, we have devised a method to directly measure the intracellular ratio of hydroquinone to quinone (HQ/Q) and have applied this measurement to correlate NQO1 enzyme abundance with HQ/Q ratio and cellular activity of IPI-504 in 30 cancer cell lines. Interestingly, the intracellular HQ/Q ratio was correlated with NQO1 levels only in a subset of cell lines and overall was poorly correlated with the growth inhibitory activity of IPI-504. Although artificial overexpression of NQO1 is able to increase the level of hydroquinone and cell sensitivity to IPI-504, it has little effect on the activity of 17-amino-17-demethoxy-geldanamycin, the major active metabolite of IPI-504. This finding could provide an explanation for the biological activity of IPI-504 in xenograft models of cell lines that are not sensitive to IPI-504 in vitro. Our results suggest that NQO1 activity is not a determinant of IPI-504 activity in vivo and, therefore, unlikely to become an important resistance mechanism to IPI-504 in the clinic. [Mol Cancer Ther 2009;8(12):3369–78]
Essential role of mitochondria in apoptosis of cancer cells induced by the marine alkaloid Lamellarin D
Authors: Ballot Caroline , Kluza Jérome , Martoriati Alain , Nyman Ulrika , Formstecher Pierre , Joseph Bertrand , Bailly Christian , Marchetti Philippe
Published: December 2009

Lamellarin D, a potent cytotoxic marine alkaloid, exerts its antitumor action through two complementary pathways: a nuclear route via topoisomerase I inhibition and a mitochondrial targeting. The present study was designed to investigate the contribution of these two pathways for apoptosis in cancer cells. Lamellarin D promoted nuclear apoptosis in leukemia cells without prominent cell cycle arrest. Signals transmitted by lamellarin D initiated apoptosis via the intrinsic apoptotic pathway. The drug induced conformational activation of Bax and decreased the expression levels of antiapoptotic proteins Bcl-2 and cIAP2 in association with activation of caspase-9 and caspase-3. Upon lamellarin D exposure, Fas and Fas-L expression was not modified in leukemia cells. Moreover, leukemia cells deficient in caspase-8 or Fas-associated protein with death domain underwent apoptosis through the typical mitochondrial apoptotic cascade, indicating that cell death induced by lamellarin D was independent of the extrinsic apoptotic pathway. Lamellarin D also exerted a topoisomerase I–mediated DNA damage response resulting in H2AX phosphorylation, and the upregulation of the DNA repair protein Rad51 and of p53, as well as the phosphorylation of p53 at serine 15. However, lamellarin D killed efficiently mutated p53 or p53 null cancer cells, and sensitivity to lamellarin D was abrogated neither by cycloheximide nor in enucleated cells. Lamellarin D–induced cytochrome c release occurs independently of nuclear factors in a cell-free system. These results suggest that lamellarin D exerts its cytotoxic effects primarily by inducing mitochondrial apoptosis independently of nuclear signaling. Thus, lamellarin D constitutes a new proapoptotic agent that may bypass certain forms of apoptosis resistance that occur in tumor cells. [Mol Cancer Ther 2009;8(12):3307–17]
COMP-Ang1 Potentiates the Antitumor Activity of 5-Fluorouracil by Improving Tissue Perfusion in Murine Lewis Lung Carcinoma
Authors: Hwang Jeong-Ah , Lee Eun Hui , Kim Hyun-Woo , Park Jin Bong , Jeon Byeong Hwa , Cho Chung-Hyun
Published: December 2009

Chemotherapy is often hindered by abnormal tumor vascularity, which causes impaired delivery of drugs into the tumor. Angiopoietin-1 has potent roles in angiogenesis and vessel maturation. We report here that an angiopoietin-1 variant, COMP-Ang1, promotes vascular remodeling and pericyte coverage and thereby promotes the efficient delivery of a chemotherapeutic drug into tumors of murine Lewis lung carcinoma. The combination of COMP-Ang1 with the cytotoxic drug 5-fluorouracil potentiated the effect of 5-fluorouracil on tumor growth without increasing animal toxicity. Moreover, COMP-Ang1 increased perfusion into the tumor. Although COMP-Ang1 increased the functional vasculature in the tumor, COMP-Ang1 alone did not promote tumor growth, possibly due to its promotion of increased pericyte coverage. This study suggests that COMP-Ang1 may improve the microcirculation within a tumor by increasing functional vasculature and tissue perfusion and that the combination of chemotherapy together with COMP-Ang1 might be an advantageous therapeutic approach. (Mol Cancer Res 2009;7(12):1920–7)
A DNA Methyltransferase Inhibitor andAll-transRetinoic Acid Reduce Oral Cavity Carcinogenesis Induced by the Carcinogen 4-Nitroquinoline 1-Oxide
Authors: Tang Xiao-Han , Albert Martin , Scognamiglio Theresa , Gudas Lorraine J.
Published: December 2009

The transcriptional silencing of some cell cycle inhibitors and tumor suppressors, such as p16 and retinoic acid receptor β2, by DNA hypermethylation at CpG islands is commonly found in human oral squamous carcinoma cells. We examined the effects of the DNA methyltransferase inhibitor 5-Aza-2′-deoxycytidine (5-Aza; 0.25 mg/kg body weight), all-trans retinoic acid (RA; given at 100 μg/kg body weight and 1 mg/kg body weight), and the combination of 5-Aza and the low-dose RA on murine oral cavity carcinogenesis induced by the carcinogen 4-nitroquinoline 1-oxide (4-NQO) in a mouse model. All the drug treatments were done for 15 weeks after a 10-week 4-NQO treatment. Mice in all drug treatment groups showed decreases in the average numbers of neoplastic tongue lesions. The combination of 5-Aza and RA effectively attenuated tongue lesion severity. Although all drug treatments limited the increase in the percentage of proliferating cell nuclear antigen–positive cells and the decrease in the percentage of p16-positive cells caused by the 4-NQO treatment in mouse tongue epithelial regions without visible lesions and in the neoplastic tongue lesions, the combination of 5-Aza and RA was the most effective. Collectively, our results show that the combination of a DNA demethylating drug and RA has potential as a strategy to reduce oral cavity cancer in this 4-NQO model.
Gender Modifies the Effect of Ursodeoxycholic Acid in a Randomized Controlled Trial in Colorectal Adenoma Patients
Authors: Thompson Patricia A. , Wertheim Betsy C. , Roe Denise J. , Ashbeck Erin L. , Jacobs Elizabeth T. , Lance Peter , Martínez María Elena , Alberts David S.
Published: December 2009

Purpose: Ursodeoxycholic acid (UDCA) was one of the earliest agents investigated as a drug for colorectal cancer prevention. However, UDCA failed to show efficacy to prevent the development of colorectal adenomas in a large, phase III, randomized, placebo-controlled trial. We re-evaluated the effect of UDCA in men and women separately, based on sex-specific differences in bile acid metabolism and suspected variation in etiologic factors contributing to colorectal cancer risk.

Experimental Design: We conducted a secondary analysis of the efficacy of UDCA to prevent colorectal adenoma in men (n = 804) and women (n = 388).

Results: We found no reduction in risk of any metachronous adenoma with UDCA treatment in men or women. However, UDCA treatment significantly lowered the odds of advanced lesions [odds ratio (OR), 0.62; 95% confidence interval (CI), 0.43-0.89] in men, but not women. We also observed significantly higher odds of advanced lesions with UDCA treatment in women who were younger (age, <65 years; OR, 3.24; 95% CI, 1.10-9.56), obese (body mass index, ≥30 kg/m2; OR, 5.45; 95% CI, 1.42-20.9), or in the highest tertile of total dietary fat (≥56.2 g/day; OR, 3.48; 95% CI, 1.35-8.95). In a multivariate model, the interactive effect of fat intake accounted for the modulating effects of age and body mass index in women.

Conclusion: Our findings support the use of UDCA for preventing advanced colorectal adenomas in men. The increased odds of adenoma among women with high fat intake suggest a previously unrecognized harm that warrants further study, especially given the chronic exposure to UDCA in patients with primary biliary cirrhosis and the increasing investigational use of UDCA for several other conditions.
Multiplex Analysis of Toll-Like Receptor-Stimulated Neonatal Cytokine Response
Authors: Caron Justin E. , La Pine Timothy R. , Augustine Nancy H. , Martins Thomas B. , Hill Harry R.
Published: December 2009

Background: The human neonate’s increased susceptibility to bacterial infections is not completely understood. Toll-like receptors (TLRs) have been recognized as pattern-recognition receptors critical to the innate immune response. TLR function in neonates, however, remains incompletely defined. Objective: To examine regulatory and proinflammatory cytokine responses to TLR-1–6 stimulation of cord blood compared to adult blood. Methods: We stimulated cord blood with ligands for each of TLRs 1–6 and compared these responses to adult controls. The following TLR ligands were utilized: Pam3CSK4 (TLR-1 and 2), zymosan (TLR-2 and 6), poly I:C (TLR-3), LPS (TLR-4), and flagellin (TLR-5). Cytokine production was measured with an assay developed in-house utilizing multi-analyte technology. Results: TLR-1–6 stimulation produced higher concentrations of proinflammatory cytokines (IL-1β, IL-6, and IL-8) in cord blood compared to adult blood, with the exception of TLR-4-stimulated TNF-α production, which was significantly lower in cord blood (319 pg/ml) compared to adult blood (645 pg/ml; p = 0.027). In contrast, TLR-1–6 stimulation resulted in decreased concentrations of Th1 and Th2 cytokines in cord blood compared to adult blood, with significantly diminished production of IL-12 (TLRs 1/2, 2/6, 3 and 4), IL-13 (TLR-1–6), and IL-10 (TLR-4). Conclusion: Cord blood production of regulatory Th1 and Th2 cytokines following TLR stimulation is decreased compared to that of adult blood. In contrast, TLR-stimulated proinflammatory cytokine production was markedly higher in neonates than in adults, with the exception of TLR-4-induced TNF-α production. The human neonate’s increased susceptibility to bacterial infections may be related to abnormal TLR responsiveness, with enhanced proinflammatory and decreased regulatory cytokine production.
Clinical Features and Treatment Outcomes of Non-Hodgkin’s Lymphomas Involving Rare Extranodal Sites: A Single-Center Experience
Authors: Yun Jina , Kim Seok Jin , Kim Jung A , Kong Jee Hyun , Lee Soo Hyeon , Kim Kihyun , Ko Young Hyeh , Kim Won Seog
Published: December 2009

Background: The involvement of certain organs such as the adrenal gland and ovaries is rare in non-Hodgkin’s lymphoma (NHL). There are few studies comparing clinical features and prognosis based on the extranodal organ involved. Methods: We selected patients presenting with predominantly extranodal involvement among patients diagnosed with NHL from 1998 to 2009 at the Samsung Medical Center. Results: Forty-eight patients with NHL involving rare extranodal sites were analyzed. The extranodal sites were as follows: adrenal gland (n = 14), ovary (n = 13), pancreas (n = 11), uterus (n = 4), esophagus (n = 4) and prostate (n = 2). Diffuse large B-cell lymphoma (DLBCL) was the most common (n = 39), and the median overall survival (OS) was 16.63 months. There was no significant difference in OS according to the involved sites. Rituximab plus CHOP failed to provide an additive survival benefit over CHOP alone in 39 DLBCL patients. The OS of DLBCL in rare extranodal sites was worse than that in common sites when compared based on tumor stage. Conclusions: NHL involving rare extranodal sites had a poor prognosis, and the impact of rituximab on survival was negligible. Thus, more intensive therapeutic strategies should be considered for NHL involving rare extranodal sites.
Connective Tissue Growth Factor-(CTGF, CCN2) – A Marker, Mediator and Therapeutic Target for Renal Fibrosis
Authors: Phanish Mysore K. , Winn S.K. , Dockrell M.E.C.
Published: December 2009

Connective tissue growth factor (CTGF, CCN2) is a key mediator of tissue fibrosis. CCN2 plays an important role in the development of glomerular and tubulointerstitial fibrosis in progressive kidney diseases. In this review, we discuss the biology of CCN2 with a focus on the regulation of CCN2 gene, cellular mechanisms of profibrotic CCN2 effects and the current in vivo and in vitro evidence for the role of CCN2 in the development of renal fibrosis. We also discuss the therapeutic potential of targeting CCN2 for the treatment of renal fibrosis.
Poor Cell Survival Limits the Beneficial Impact of Mesenchymal Stem Cell Transplantation on Acute Kidney Injury
Authors: Burst Volker R. , Gillis Meyke , Pütsch Florian , Herzog Ruth , Fischer Jürgen H. , Heid Petra , Müller-Ehmsen Jochen , Schenk Kerstin , Fries Jochen W.U. , Baldamus Conrad A. , Benzing Thomas
Published: December 2009

Background: Although renal tubular epithelium has a great capacity for repair it has been suggested that the administration of mesenchymal stem cells may accelerate the recovery following severe ischemic injury. Methods: Here we analyzed the survival rate and organ distribution of transplanted mesenchymal stem cells as well as their contribution to kidney regeneration after ischemic renal injury using functional tests, histological examination as well as quantitative real-time PCR. Results: Intravenously injected stem cells were mainly trapped in lungs and liver. One hour after injection, less than 1% of the injected stem cells could be detected in the injured kidneys. These cells disappeared within the first few days and did not replace renal epithelial cells precluding substantial transdifferentiation. To clarify whether reinforced stem cell delivery might promote sustained survival or conversion to tubular epithelia, stem cells were directly injected into the injured kidneys. Although these grafted cells also did not show sustained survival or contribute to structural renal repair, stem cell injection was associated with a significant but transient initial decrease in serum creatinine. Conclusion: These data suggest that mesenchymal stem cells do not significantly contribute to epithelial renewal after ischemic injury, promoting the idea that the major impact of cell-based therapy for acute kidney injury may result from paracrine or endocrine effects unrelated to stem cell transdifferentiation.
Pattern of Bone Mineral Density in Sickle Cell Disease Patients with the High-Hb F Phenotype
Authors: Gupta Renu , Marouf Rajaa , Adekile Adekunle
Published: December 2009

Low bone mineral density (BMD) is common in sickle cell disease (SCD) patients of all ages due to erythropoietic stress and consequent bone marrow hyperplasia. Kuwaiti SCD patients have a mild clinical phenotype because of their high Hb F level. There has been no previous documentation of BMD in this population of patients. SCD patients (n = 81) and controls (n = 67) were recruited from hematology clinics of Mubarak Hospital, Kuwait. BMD was measured using dual X-ray absorptiometry (Hologic) in the lumbar spine and the hip (left femoral neck). Among the 53 adult patients, the prevalence of low BMD was 67.4% in the spine and 33.3% in the hip while among controls, the figures were 23.1 and 11.3%, respectively. These differences are significant (p < 0.01). In SCD children, the figures were 17.9% (in the lumbar spine) and 3.6% (in the hip), while in controls the figures were 13.3 and 0%, respectively. The differences are not significant (p > 0.05). Patients with frequent vaso-occlusive crisis had significantly lower mean BMD, but those with MRI evidence of avascular necrosis of the femoral head were more likely to have normal or osteosclerotic BMD. Our study showed that osteopenia/osteoporosis is uncommon among Kuwaiti children with SCD but quite common in adults.
Shock Wave Therapy Effectively Attenuates Inflammation in Rat Carotid Artery following Endothelial Denudation by Balloon Catheter
Authors: Shao Pei-Lin , Chiu Chaw-Chi , Yuen Chun-Man , Chua Sarah , Chang Li-Teh , Sheu Jiunn-Jye , Sun Cheuk-Kwan , Wu Chiung-Jen , Wang Ching-Jen , Yip Hon-Kan
Published: December 2009

Background: This study investigates the effectiveness of extracorporeal shock wave (ECSW) in ameliorating inflammatory mediator expression and neointimal formation in a rat model of vascular injury. Methods and Results: Male Sprague-Dawley rats with left carotid artery (LCA) injury induced by balloon dilatation (BD; group 1) were compared with group 2 [LCA injury plus ECSW-181 (defined as 181 total shocks given in LCA at 0.011 mJ/mm2) on day 2 post-LCA injury], and group 3 (normal controls). The rats in each group were further divided into 3 subgroups (n = 6, each) that were sacrificed on postoperative day 3, 7 and 14, respectively. The results demonstrated that, compared to groups 2 and 3, group 1 had significantly increased cellular expression of CD40, interleukin-18, and connexin 43 at each analyzed time point (all p < 0.001). Additionally, LCCA macrophage (CD68) recruitment was substantially increased in group 1 compared to groups 2 and 3 (all p < 0.001). Furthermore, LCA neointimal proliferation and media thickness were markedly higher in group 1 than in groups 2 and 3 on days 7 and 14 post-BD (all p < 0.001). Conclusions: ECSW markedly attenuates inflammatory responses, proliferation of neointima and smooth muscle cells in a rat vascular injury model.
CD9 Gene Variations Are Not Associated with Female Infertility in Humans
Authors: Nishiyama Sachie , Kishi Taro , Kato Takema , Suzuki Machiko , Nishizawa Haruki , Pryor-Koishi Kanako , Sawada Tomio , Nishiyama Yukio , Iwata Nakao , Udagawa Yasuhiro , Kurahashi Hiroki
Published: December 2009

Background/Aims: To determine whether genetic alterations in the CD9 gene are associated with female infertility in humans. Methods: We sequenced the entire coding region of this gene in 86 Japanese women with unexplained infertility and further conducted a case-control study of six tagging single nucleotide polymorphisms (SNPs) in this gene using an additional 164 samples obtained from a fertile control group. Results: No disease-causing mutation in the CD9 gene was evident in these samples and no significant association between the tagging SNPs and the studied cohort was identified. Conclusions: Our findings do not support the hypothesis that genetic alterations of the CD9 gene cause female infertility in humans.
Expression of FXYD3 Protein in Relation to Biological and Clinicopathological Variables in Colorectal Cancers
Authors: Widegren Emma , Önnesjö Sofia , Arbman Gunnar , Kayed Hany , Zentgraf Hanswalter , Kleeff Jörg , Zhang Hong , Sun Xiao-Feng
Published: December 2009

Background: FXYD3 is up-/down-regulated in different types of cancers. We examined FXYD3 expression in colorectal cancers and its relationship to biological and clinicopathological variables. Patients and Methods: Expression of FXYD3 protein was immunohistochemically examined in distant normal mucosa (n = 34), adjacent normal mucosa (n = 72), primary tumour (n = 150) and lymph node metastasis (n = 35) from colorectal cancer patients. Results: FXYD3 was highly expressed in primary tumour compared to adjacent normal mucosa (p = 0.02). FXYD3 was or tended to be positively related to the expression of ras (p = 0.02), p53 (p = 0.06), legumain (p = 0.02) and proliferating cell nuclear antigen (p = 0.03). Moreover, there was a higher frequency of strong FXYD3 expression in Dukes A–C tumours than in D tumours (p = 0.04). The strong FXYD3 expression tended to predict worse survival in the patients with Dukes A + B tumour (p = 0.07), while there was no such tendency in the patients with Dukes C + D tumour (p = 0.94). The tumours located in the colon had a higher degree of FXYD3 expression than the tumours located in the rectum (p = 0.05). Conclusion: The FXYD3 was associated with certain biological variables and may be involved in the development of the relative earlier stages of colorectal cancers.
Benzoxazine Series of Histidine Kinase Inhibitors as Potential Antimicrobial Agents with Activity against Enterococci
Authors: Mueller-Premru Manica , Zidar Nace , Cvitković Špik Vesna , Krope Adrijana , Kikelj Danijel
Published: December 2009

Background: Antibacterial activity of three members of a benzoxazine series of histidine kinase inhibitors (4a, 4b and 6) was studied on standard strains, and with 4b also on clinical isolates of enterococci. Methods: Susceptibility to each compound was tested using a broth macrodilution method with a range of dilutions from 0.016 to 128 μg/ml on four standard strains (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 29213 and Enterococcus faecalis ATCC 29212) and in the case of 4b also on 52 clinical isolates of enterococci (7 vancomycin-susceptible E. faecalis, 15 vancomycin-susceptible Enterococcus faecium and 30 vancomycin-resistant E. faecium), and on two additional standard strains (S. aureus ATCC 43300 and E. faecalis ATCC 51299). Results: Neither of the compounds inhibited the growth of E. coli ATCC 25922 or P. aeruginosa ATCC 27853. Compound 4b inhibited the growth of S. aureus ATCC 29213 at 8 μg/ml, S. aureus ATCC 43300 at 32 μg/ml, E. faecalis ATCC 29212, and E. faecalis ATCC 51299 at 16 μg/ml, while 4a and 6 did so at higher concentrations. For clinical isolates of enterococci, the minimal inhibitory concentrations of 4b were in the range of 0.5–16 μg/ml. Conclusions: Compound 4b promises to be a potentially useful antimicrobial agent for vancomycin-susceptible and -resistant enterococci.
Low Erythropoietin Plasma Levels during Exacerbations of COPD
Authors: Sala Ernest , Balaguer Catalina , Villena Cristina , Ríos Angel , Noguera Aina , Núñez Belén , Agustí Alvar
Published: December 2009

Background: It is known that pro-inflammatory cytokines suppress in vitro the gene expression and protein production of erythropoietin (Epo). We hypothesized that systemic inflammation in patients with chronic obstructive pulmonary disease (COPD) may influence Epo production, particularly during episodes of exacerbation of the disease (ECOPD) where an inflammatory burst is known to occur. Objectives: We compared the plasma levels of Epo and high-sensitivity (hs) C-reactive protein (hsC-RP) in patients hospitalized because of ECOPD (n = 26; FEV1: 48 ± 15% predicted), patients with clinically stable COPD (n = 31; FEV1: 49 ± 17% predicted), smokers with normal lung function (n = 9), and healthy never smokers (n = 9). Methods: Venous blood samples were taken between 9 and 10 a.m. after an overnight fast into tubes with EDTA (10 ml) or without EDTA (10 ml). Plasma levels of Epo (R&D Systems Inc., Minneapolis, Minn., USA) and hsC-RP (BioSource, Belgium) were determined by ELISA. Results: Log-Epo plasma levels were significantly lower (0.46 ± 0.32 mU/ml) in ECOPD than in stable COPD (1.05 ± 0.23 mU/ml), smokers (0.95 ± 0.11 mU/ml) and never smokers with normal lung function (0.92 ± 0.19 mU/ml) (p < 0.01, each). In a subset of 8 COPD patients who could be studied both during ECOPD and clinical stability, log-Epo increased from 0.49 ± 0.42 mU/ml during ECOPD to 0.97 ± 0.19 mU/ml during stability (p < 0.01). In patients with COPD log-Epo was significantly related to hsC-RP (r = –0.55, p < 0.0001) and circulating neutrophils (r = –0.48, p < 0.0001). Conclusions: These results show that the plasma levels of Epo are reduced during ECOPD likely in relation to a burst of systemic inflammation.
Premalignant Circumscribed Palmar Hypokeratosis: A New Form of Circumscribed Palmar Hypokeratosis? Case Report and Literature Review
Authors: Kanitakis J. , Lora V. , Balme B. , Roby J.
Published: December 2009

Circumscribed palmoplantar hypokeratosis (CPH) is a recently described dermatosis of unknown origin, of which 46 cases have been published so far. CPH affects predominantly adult women. It presents clinically with a sharply circumscribed annular erythematous depressed plaque rimmed by a slightly hyperkeratotic border, localized usually over the thenar or hypothenar eminences of the palm, less commonly on the soles. Histopathologically, it is characterized by an abrupt decrease in the horny layer thickness, forming a sharp stair between normal and involved skin. We report herein a new patient with CPH who presented an associated aspect of actinic keratosis, a hitherto unreported finding. This case suggests that CPH can undergo malignant transformation, probably as a result of actinic damage. On this occasion, a complete literature review of CPH is presented.