bluuebunny/crossref_metadata_2025 huggingface dataset:
Preventing microalbuminuria in patients with type 2 diabetes
Authors: Jermendy György , Ruggenenti Piero
Published: November 2006

The public health burden of type 2 diabetes mellitus has been dramatically increasing world‐wide. The chronic complications of type 2 diabetes play an important role in decreasing life expectancy and adversely affecting quality of life. Diabetic nephropathy, which is originally microvascular in nature, is widely considered an important complication of diabetes. In prospective clinical investigations, increased urinary albumin excretion proved to be associated not only with subsequent renal outcomes but also with cardiovascular morbidity/mortality independently of other risk factors. Therefore, microalbuminuria as an early sign of increased urinary albumin excretion should be considered important for both treatment and even for prevention. Preventing microalbuminuria might diminish progression to overt nephropathy and, hopefully, might limit cardiovascular events. Regarding primary prevention of diabetic nephropathy, therapeutic intervention should optimally be initiated at the stage of normoalbuminuria. Although additional factors such as smoking cessation, reduction of protein intake, and treatment of lipid abnormalities are important, providing optimal diabetic control as well as targeting optimal blood pressure are the key elements of a prevention strategy in diabetic patients. Recently, the Bergamo Nephrologic Diabetes Complications Trial (BENEDICT) documented that a significant decrease of the development of persistent microalbuminuria could be achieved by using an ACE‐inhibitor, trandolapril alone or in combination with verapamil SR, a non‐dihydropyridine calcium‐channel blocker in hypertensive type 2 diabetic patients with normoalbuminuria. The results of this primary‐prevention strategy should be corroborated by further investigations to determine whether these beneficial changes could later result in improvement of renal clinical outcomes, macrovascular complications, or both. Copyright © 2006 John Wiley & Sons, Ltd.
Atoh1 expression defines activated progenitors and differentiating hair cells during avian hair cell regeneration
Authors: Cafaro Jon , Lee Gi Soo , Stone Jennifer S.
Published: November 2006

In the avian inner ear, nonsensory supporting cells give rise to new sensory hair cells through two distinct processes: mitosis and direct transdifferentiation. Regulation of supporting cell behavior and cell fate specification during avian hair cell regeneration is poorly characterized. Expression of Atoh1, a proneural transcription factor necessary and sufficient for developmental hair cell specification, was examined using immunofluorescence in quiescent and regenerating hair cell epithelia of mature chickens. In untreated birds, Atoh1 protein was not detected in the auditory epithelium, which is quiescent. In contrast, numerous Atoh1‐positive nuclei were seen in the utricular macula, which undergoes continual hair cell turnover. Atoh1‐positive nuclei emerged in the auditory epithelium by 15 hr post‐ototoxin administration, before overt hair cell damage and supporting cell re‐entry into the cell cycle. Subsequently, Atoh1 labeling was seen in 15% of dividing supporting cells. During cell division, Atoh1 was distributed symmetrically to daughter cells, but Atoh1 levels were dramatically regulated shortly thereafter. After cellular differentiation, Atoh1 labeling was confined to hair cells regenerated through either mitosis or direct transdifferentiation. However, Atoh1 expression in dividing progenitors did not necessarily predict hair cell fate specification in daughter cells. Finally, predominant modes of hair cell regeneration varied significantly across the radial axis of the auditory epithelium, with mitosis most frequent neurally and direct transdifferentiation most frequent abneurally. These observations suggest a role for Atoh1 in re‐specifying supporting cells and in biasing postmitotic cells toward the hair cell fate during hair cell regeneration in the mature chicken ear. Developmental Dynamics 236:156–170, 2007. © 2006 Wiley‐Liss, Inc.
Reactions of GaCp* with a Hemilabile Derivative of Rh6(CO)16 – Synthesis and Structural Characterization of Two Novel Heterometallic Clusters: Rh6(CO)13(μ,κ3‐Ph2PC2H3)(μ3‐GaCp*) and Rh6(CO)13(κ1‐Ph2PC2H3)(μ3‐GaCp*)2
Authors: Grachova Elena V. , Linti Gerald , Stammler Hans‐Georg , Neumann Beate , Tunik Sergey P. , Wadepohl Hubert
Published: November 2006

The reaction of Rh6(CO)14(Ph2PCH=CH2) with a twofold excess of GaCp* (Cp* = C5Me5) affords two novel heterometallic clusters Rh6(CO)13(μ,κ3‐Ph2PC2H3)(μ3‐GaCp*) (1) and Rh6(CO)13(κ1‐Ph2PC2H3)(μ3‐GaCp*)2 (2) in a moderate yield. The hemilabile behavior of coordinated diphenylvinylphosphane in the starting cluster allows the easy insertion of GaCp* into the cluster coordination sphere, which eventually gives the cluster 1 with face‐bridging coordination of the gallium fragment and the phosphane ligand recoordinated in an edge‐bridging position. The further substitution of the vinyl function in 1 for the second GaCp* ligand affords cluster 2, where recoordination of the double bond is blocked by steric hindrance from adjacent GaCp* fragments. Both clusters were structurally characterized by X‐ray crystallography in the solid state and by IR and NMR spectroscopy in solution. Hemilability of diphenylvinylphosphane in 1 was independently confirmed by the cluster reaction with CO, which gives the Rh6(CO)14(κ1‐Ph2PC2H3)(GaCp*) adduct (3) with the phosphane ligand only coordinated through the phosphorus atom. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2007)
A Water Pipe Held Up by a Polyoxometalate Supported Transition Metal Complex: Synthesis and Characterization of [Cu2(phen)2(CH3COO)(CH3COOH)(H2O)2][Al(OH)6Mo6O18]·28H2O
Authors: Shivaiah Vaddypally , Chatterjee Tanmay , Srinivasu Khandregula , Das Samar K.
Published: November 2006

A water pipe, formed exclusively from lattice water molecules, is stabilized in an organic–inorganic hybrid materialCu2(phen)2(CH3COO)(CH3COOH)(H2O)2][Al(OH)6Mo6O18]·28H2O (1). Compound 1 consists of an Anderson‐type heteropolyanion, a copper acetate dimer, and 28 lattice water molecules. Once the relevant crystal is removed from the mother liquor it loses water molecules, which results in the demolition of the water pipe. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2007)
Synthesis of 3‐Methoxy‐16α‐nitro‐14,17‐ethenoestra‐1,3,5(10)‐trien‐17β‐yl Acetate and Fragmentation‐Mediated Pathways to 14β,15β‐Fused N‐Heterocycles and 14β‐Functionalised Alkyl Derivatives
Authors: Baranovsky Alexander V. , Bolibrukh Dmitry A. , Bull James R.
Published: November 2006

The Diels–Alder reaction of 3‐methoxyestra‐1,3,5(10),14,16‐pentaen‐17‐yl acetate (1) with nitroethylene gives 3‐methoxy‐16‐nitro‐14,17‐ethenoestra‐1,3,5(10)‐trien‐17β‐yl acetate (2). Cycloadduct 2 can be reduced with TiCl3, giving oxazine 3 as a result of sigmatropic rearrangement, whereas the treatment of 2 with Lewis acids (TiCl4, SnCl4) leads to a derivative of the cyclic hydroximic acid 10; the latter is viewed as a product of intramolecular rearrangement. Cycloadduct 2 suffers a weak base‐induced cleavage of the C(16)–C(17) bond, releasing the nitrile oxide intermediate, which can be trapped by a dipolarophile or reduced with triphenylphosphane. Thus, the trapping of the nitrile oxide with propargyl alcohol gives the isoxazole 12 in 50 % yield, and the reduction of the nitrile oxide with triphenylphosphane gives nitrile 9 in 94 % yield. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2007)
Synthesis of an Enantiomerically Pure Building Block for the Synthesis of Hydroporphyrins
Authors: Etornam Adukpo Genevieve , Borrmann Tobias , Manski René , Sáez Díaz Rosa I. , Stohrer Wolf‐Dieter , Montforts Franz‐Peter
Published: November 2006

The enantiomerically pure pyrrolidine diester 4 is a useful building block for the synthesis of chiral hydroporphyrin compounds. Treatment of optically active aromatic amines with bislactone 5 gave pairs of N‐alkylated lactam‐lactone diastereomers 6 and 8. These diastereomers were separated by MPL chromatography and in the case of 8 they could be debenzylated to yield the enantiomerically pure lactam‐lactone 7 and its enantiomer. The (–)‐lactam‐lactone enantiomer 7 was further transformed into building block 4.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2007)
A Novel C3‐Symmetric Triol as Chiral Receptor for Ammonium Ions
Authors: Fabris Fabrizio , Pellizzaro Leonardo , Zonta Cristiano , De Lucchi Ottorino
Published: November 2006

The enantiopure C3‐symmetric syn‐benzotriborneol was efficiently obtained in an eight‐step route. Of particular interest from a synthetic point of view are the potassium tert‐butoxide promoted bromination of camphor hydrazone (4) and the observation that the protecting groups influence the syn/anti diasteroselectivity of the cyclotrimerization reaction of 12–14. Complexation studies in deuteriochloroform revealed the capability of the cyclotrimer syn‐1 to act as host for ammonium ions, and in particular, the efficient chiral recognition of the two enantiomers of (1‐phenylethyl)ammonium chloride. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2007)
Bruder, Broder, Brauder – Dialekte in Deutschland
Authors: Lameli Alfred , Rabanus Stefan
Published: November 2006

Flächendeckend und präzise dokumentiert der „Sprachatlas des Deutschen Reiches”︁ die Dialekte des 19. Jahrhunderts. Die vollständige Digitalisierung der Sprachkarten eröffnet der internationalen Forschung neue Möglichkeiten
Das Liebesleben der Zebrafinken
Authors: Unterstell Rembert
Published: November 2006

Der Verhaltensgenetiker Wolfgang Forstmeier studiert die Grundlagen und Konsequenzen sexuellen Verhaltens
Über „Knäuelpilze”︁ und Nebelwald
Authors: Kottke Ingrid , Nebel Martin
Published: November 2006

In den Anden leben Pflanzen in Symbiose mit Wurzelpilzen. Über die Netzwerke der Pilze im Boden wird die Artenvielfalt im Ökosystem gefördert
Power calculations for likelihood ratio tests for offspring genotype risks, maternal effects, and parent‐of‐origin (POO) effects in the presence of missing parental genotypes when unaffected siblings are available
Authors: Rampersaud E. , Morris R.W. , Weinberg C.R. , Speer M.C. , Martin E.R.
Published: November 2006

Genotype‐based likelihood‐ratio tests (LRT) of association that examine maternal and parent‐of‐origin effects have been previously developed in the framework of log‐linear and conditional logistic regression models. In the situation where parental genotypes are missing, the expectation‐maximization (EM) algorithm has been incorporated in the log‐linear approach to allow incomplete triads to contribute to the LRT. We present an extension to this model which we call the Combined_LRT that incorporates additional information from the genotypes of unaffected siblings to improve assignment of incompletely typed families to mating type categories, thereby improving inference of missing parental data. Using simulations involving a realistic array of family structures, we demonstrate the validity of the Combined_LRT under the null hypothesis of no association and provide power comparisons under varying levels of missing data and using sibling genotype data. We demonstrate the improved power of the Combined_LRT compared with the family‐based association test (FBAT), another widely used association test. Lastly, we apply the Combined_LRT to a candidate gene analysis in Autism families, some of which have missing parental genotypes. We conclude that the proposed log‐linear model will be an important tool for future candidate gene studies, for many complex diseases where unaffected siblings can often be ascertained and where epigenetic factors such as imprinting may play a role in disease etiology.Genet. Epidemiol. © 2006 Wiley‐Liss, Inc.
Chronic transplantation of olfactory ensheathing cells promotes partial recovery after complete spinal cord transection in the rat
Authors: López‐Vales Rubèn , Forés Joaquim , Navarro Xavier , Verdú Enrique
Published: November 2006

The goal of this study was to ascertain whether olfactory ensheathing cells (OECs) were able to promote axonal regeneration and functional recovery when transplanted 45 days after complete transection of the thoracic spinal cord in adult rats. OECs promoted partial restitution of supraspinal pathways evaluated by motor evoked potentials and modest recovery of hindlimb movements. In addition, OEC grafts reduced lumbar reflex hyperexcitability from the first month after transplantation. Histological results revealed that OECs facilitated corticospinal and raphespinal axons regrowth through the injury site and into the caudal spinal cord segments. Interestingly, raphespinal but not corticospinal fibers regenerated long distances through the gray matter and reached the lower lumbar segments (L5) of the spinal cord. However, delayed OEC grafts failed to reduce posttraumatic astrogliosis. In conclusion, the beneficial effects found in the present study further support the use of OECs for treating chronic spinal cord injuries. © 2006 Wiley‐Liss, Inc.
Adenovirus vaccination against neu oncogene exerts long‐term protection from tumorigenesis in BALB/neuT transgenic mice
Authors: Gallo Pasquale , Dharmapuri Sridhar , Nuzzo Maurizio , Maldini Daniele , Cipriani Barbara , Forni Guido , Monaci Paolo
Published: November 2006

The transforming rat HER2/neu oncogene (neu), when embedded in the genome of transgenic BALB/c (neuT) mice, provokes the development of an invasive carcinoma in each of their 10 mammary glands. We used the neuT mice model system to evaluate the immunization efficiency and the protective effect of intramuscular injection of adenovirus (Ad) and/or of DNA with electrostimulation (DNA+ES), both expressing the rat p185neu protein. A neu cDNA sequence, which exclusively contains codons preferred by highly expressed mammalian genes, was used in this study. This “optimized” cDNA displayed higher expression in cultured cells and greater cell‐mediated response than the original gene when injected as DNA+ES. Ad expressing the optimized sequence (Ad5‐neu.opt) induced a higher immune response, as measured by the frequency of IFN‐γ‐secreting spleen cells and antibody titers. Different Ad/DNA combinations and immunization schedules confirmed the superiority of Ad5‐neu.opt in inducing a strong Th1‐skewed humoral and CD8+ cell‐mediated response. Two Ad5‐neu.opt injections of 109 viral particles at week 10 and 12 were sufficient to induce the highest response, which persisted at detectable levels up to 33 weeks of age. Anti‐Ad5 antibodies elicited by previous injections neutralized the effect of an additional Ad5‐neu.opt immunization at week 19. A group, which received 3 injections of DNA+ES at week 23, 27 and 31, in addition to the 3 Ad injections at week 10, 12 and 19 showed an increased frequency of IFN‐γ+, CD8+ PBMC at week 25, which persisted at detectable levels till week 38. Ad5‐neu.opt administration at 10 and 12 weeks of age had a significant impact on tumor progression. At 44 weeks, 40% of the mice were completely protected from tumors with a mean tumor of 3.8. In contrast, control mice developed 10 tumors and died by week 27. Vaccination blocked the tumor development at the atypical hyperplasia stage present at the time of treatment. Tumors developing at later times express reduced levels of rat p185neu protein. © 2006 Wiley‐Liss, Inc.
Successful cancer vaccine therapy for carcinoembryonic antigen (CEA)‐expressing colon cancer using genetically modified dendritic cells that express CEA and T helper‐type 1 cytokines in CEA transgenic mice
Authors: Ojima Toshiyasu , Iwahashi Makoto , Nakamura Masaki , Matsuda Kenji , Nakamori Mikihito , Ueda Kentaro , Naka Teiji , Ishida Koichiro , Primus F. James , Yamaue Hiroki
Published: November 2006

This study was designed to determine whether the vaccination of genetically modified dendritic cells (DCs) simultaneously expressing carcinoembryonic antigen (CEA), granulocyte macrophage colony‐stimulating factor (GM‐CSF) and interleukin 12 (IL‐12) can overcome the peripheral T‐cell tolerance to CEA and thereby elicit a therapeutic response in CEA transgenic mice. CEA transgenic mice were immunized once by subcutaneous injection with DCs adenovirally transduced with CEA and T helper‐type 1 cytokine genes. The cytotoxic activity of spleen cells against CEA‐expressing tumors, MC38‐CEA, in the mice immunized with DCs expressing CEA (DC‐AxCACEA) was higher than that in those immunized with DCs‐AxCALacZ (p < 0.0001), and was augmented by the cotransduction with the GM‐CSF/IL‐12 gene (p < 0.05). The vaccination with DC‐AxCACEA/GM‐CSF/IL‐12 could elicit a more potent therapeutic immunity than the vaccination with DC‐AxCACEA in subcutaneous tumor models (p < 0.0001), and 4 of 5 mice showed a complete eradication of the subcutaneous tumors in these vaccination groups. Even in a large tumor model, this vaccination therapy completely eliminated the subcutaneous tumors in all mice. This antitumor activity mostly vanished with the depletion of CD8+ T cells and NK cells in vivo and was completely abrogated with the depletion of CD4+ T cells. A histopathological examination showed no evidence of an autoimmune reaction. No other adverse effects were observed. This vaccination strategy resulted in the generation of highly efficient therapeutic immune responses against MC38‐CEA in the absence of autoimmune responses and demonstrated no adverse effects, and may therefore be useful for future clinical applications as a cancer vaccine therapy. © 2006 Wiley‐Liss, Inc.
Alcohol intake and colorectal cancer risk: A dose–response meta‐analysis of published cohort studies
Authors: Moskal Aurélie , Norat Teresa , Ferrari Pietro , Riboli Elio
Published: November 2006

The epidemiologic evidence support that alcohol intake might be associated with increased colorectal cancer risk. However, the results by anatomic site in the large bowel are inconsistent. We conducted a meta‐analysis of prospective cohort studies published between 1990 and June 2005 on the relationship between alcohol intake and colon and rectal cancer. We quantified associations with colon and rectal cancer using meta‐analysis of relative risk (RR) associated to the highest versus the lowest category of alcohol intake and meta‐analysis of study‐specific dose–response slopes using fixed or random effect models depending on the heterogeneity of effects among studies. Sixteen prospective cohort studies including more than 6,300 patients with colorectal cancer were eligible for inclusion. High alcohol intake was significantly associated with increased risk of colon (RR = 1.50; 95% CI = 1.25, 1.79) and rectal cancer (RR = 1.63; 95% CI = 1.35, 1.97) when comparing the highest with the lowest category of alcohol intake, equivalent to a 15% increase of risk of colon or rectal cancer for an increase of 100 g of alcohol intake per week. The relationship did not differ significantly by anatomical site (colon, rectum). Using meta‐regression analysis, we identified geographical area where the study was conducted as a possible source of between‐study heterogeneity of effects among studies. Lifestyle recommendations for prevention of colorectal cancer should consider limiting alcohol intake. © 2006 Wiley‐Liss, Inc.
Risk and risk reduction involving arginine intake and meat consumption in colorectal tumorigenesis and survival
Authors: Zell Jason A. , Ignatenko Natalia A. , Yerushalmi Hagit F. , Ziogas Argyrios , Besselsen David G. , Gerner Eugene W. , Anton‐Culver Hoda
Published: November 2006

Elevated polyamine and nitric oxide levels (both derived from arginine) promote tumorigenesis, whereas non‐steroidal anti‐inflammatory drugs (NSAIDs) inhibit colorectal cancer (CRC) incidence in experimental and epidemiologic studies. We investigated dietary arginine‐induced intestinal tumorigenesis and NSAID‐inhibitory effects in ApcMin/+ mice differentially expressing nitric oxide synthase‐2 (Nos2). We also studied effects of estimated arginine exposures through meat consumption on tumor characteristics and survival in human CRC cases. Dietary arginine increased high‐grade colon adenoma incidence in ApcMin/+Nos2+/+ mice, but not in Nos2 knockout mice. Additionally, celecoxib suppressed intestinal steady state ornithine decarboxylase RNA levels (p < 0.001), induced steady state spermidine/spermine N1‐acetyltransferase RNA levels (p = 0.002), decreased putrescine levels (p = 0.04) and decreased tumor number in the small intestines of ApcMin/+Nos2+/+ mice (p = 0.0003). Five hundred and eleven cases from our NCI‐supported CRC gene‐environment study were analyzed based on self‐reported meat (as a surrogate for arginine) consumption. Familial CRC cases (n = 144) in the highest meat consumption quartile (Q4) had no statistically significant differences in tumor grade compared to cases in Q1–Q3 (p = 0.32); however, they were observed to have decreased overall survival (OS) (10‐year OS = 42% vs. 65%; p = 0.017), and increased risk of death in an adjusted analysis (hazards ratio [HR] = 2.24; p = 0.007). No differences in tumor grade, OS or adjusted HR were observed for sporadic CRC cases (n = 367) based on meat consumption. Our results suggest important roles for arginine and meat consumption in CRC pathogenesis, and have implications for CRC prevention. © 2006 Wiley‐Liss, Inc.
The prognostic significance of thymidylate synthase and dihydropyrimidine dehydrogenase in colorectal cancer of 303 patients adjuvantly treated with 5‐fluorouracil
Authors: Jensen Søren Astrup , Vainer Ben , Sørensen Jens Benn
Published: November 2006

Cytotoxic effect of 5‐fluorouracil 5‐FU is mediated through inhibition of thymidylate synthase (TS), and 5‐FU is catabolised by dihydropyrimidine dehydrogenase (DPD). Efficacy of 5‐FU may therefore depend on the TS and DPD activity of colorectal cancer. Archival tumour specimens from 303 consecutive patients were analysed for the expression of TS and DPD using immunohistochemistry. All patients were completely resected for colorectal cancer stages II–III and have subsequently received adjuvant treatment with 5‐FU. In a multivariate analysis adjusting for the impact of bowel obstruction and vascular tumour invasion, diffuse TS pattern was significantly associated with increased risk of recurrence (hazard ratio (HR) = 1.9; 95% confidence interval (CI): 1.1–3.2; p = 0.02), but without significant association to death (HR = 1.6; 95% CI: 0.9–2.8; p = 0.08). High TS intensity was not significantly associated with lower risk of recurrence (HR = 0.6; 95% CI: 0.3–1.1; p = 0.07) or death (HR = 0.6; 95% CI: 0.3–1.2; p = 0.2). High DPD intensity was significantly associated with increased risk of recurrence (HR = 1.5; 95% CI: 1.1–2.3; p = 0.03) and death (HR = 1.6; 95% CI: 1.1–2.5; p = 0.02). Patients with a combination of low TS and high DPD intensity were at significantly increased risk of both recurrence (HR = 2.1; 95% CI: 1.0–4.2; p = 0.04) and death (HR = 2.0; 95% CI: 1.0–4.0; p = 0.05). No relationship between tolerability and toxicity of 5‐FU and TS and DPD expression was found. It is concluded that characterizing colorectal carcinomas by TS and DPD expression may disclose subsets of patients with significantly greater risk of disease recurrence and early death. This may be utilized in the selection of patients for treatment approaches and for decision on follow‐up programs. © 2006 Wiley‐Liss, Inc.
Renal cell carcinoma does not express argininosuccinate synthetase and is highly sensitive to arginine deprivationviaarginine deiminase
Authors: Yoon Cheol‐Yong , Shim Young‐Jun , Kim Eun‐Ho , Lee Ju‐Han , Won Nam‐Hee , Kim Jeong‐Hun , Park In‐Sun , Yoon Duck‐Ki , Min Bon‐Hong
Published: November 2006

Recently, pegylated arginine deiminase (ADI; EC 3.5.3.6) has been used to treat the patients with hepatocellular carcinoma or melanoma, in which the level of argininosuccinate synthetase (ASS) activity is low or undetectable. The efficacy of its antitumor activity largely depends on the level of intracellular ASS, which enables tumor cells to recycle citrulline to arginine. Thus, we examined the expression levels of ASS in various cancer cells and found that it is low in renal cell carcinoma (RCC) cells, rendering the cells highly sensitive to arginine deprivation by ADI treatment. Immunohistochemical analysis revealed that in biopsy specimens from RCC patients (n= 98), the expression of ASS is highly demonstrated in the epithelium of normal proximal tubule but not seen in tumor cells. Furthermore, RCC cells treated with ADI showed remarkable growth retardation in a dose dependent manner. ADI also exertedin vivoantiproliferative effect on the allografted renal cell carcinoma (RENCA) tumor cells and prolonged the survival of tumor‐bearing mice. Histological examination of the tumors revealed that tumor angiogenesis and vascular endothelial growth factor (VEGF) expression were significantly diminished by ADI administration. Therefore, these findings suggest that arginine deprivation by ADI could provide a beneficial strategy for the treatment of RCC in ways of inhibitions of arginine availability and neovascularization. © 2006 Wiley‐Liss, Inc.
High resolution tiling‐path BAC array deletion mapping suggests commonly involved 3p21‐p22 tumor suppressor genes in neuroblastoma and more frequent tumors
Authors: Hoebeeck Jasmien , Michels Evi , Menten Björn , Van Roy Nadine , Eggert Angelika , Schramm Alexander , De Preter Katleen , Yigit Nurten , De Smet Els , De Paepe Anne , Laureys Geneviève , Vandesompele Jo , Speleman Frank
Published: November 2006

The recurrent loss of 3p segments in neuroblastoma suggests the implication of 1 or more tumor suppressor genes but thus far few efforts have been made to pinpoint their detailed chromosomal position. To achieve this goal, array‐based comparative genomic hybridization was performed on a panel of 23 neuroblastoma cell lines and 75 primary tumors using a tiling‐path bacterial artificial chromosome array for chromosome 3p. A total of 45 chromosome 3 losses were detected, including whole chromosome losses, large terminal deletions and interstitial deletions. The latter, observed in cell lines as well as a number of distal deletions detected in primary tumors, allowed us to demarcate 3 minimal regions of loss of 3.6 Mb [3p21.31‐p21.2, shortest regions of overlap (SRO)1], 1.4 Mb (3p22.3‐3p22.2, SRO2) and 3.8 Mb (3p25.3‐p25.1, SRO3) in size. The present data significantly extend previous findings and now firmly establish critical regions on 3p implicated in neuroblastoma. Interestingly, the 2 proximal regions coincide with previously defined SROs on 3p21.3 in more frequent tumors including lung and breast cancer. As such, similar tumor suppressor genes may play a critical role in development or progression of a variety of neoplasms, including neuroblastoma. © 2006 Wiley‐Liss, Inc.
RhoB is frequently downregulated in non‐small‐cell lung cancer and resides in the 2p24 homozygous deletion region of a lung cancer cell line
Authors: Sato Naohito , Fukui Takayuki , Taniguchi Tetsuo , Yokoyama Toshihiko , Kondo Masashi , Nagasaka Tetsuro , Goto Yasuhiro , Gao Wentao , Ueda Yuichi , Yokoi Kohei , Minna John D. , Osada Hirotaka , Kondo Yutaka , Sekido Yoshitaka
Published: November 2006

Identification of a homozygous deletion in cancer cells provides strong evidence for the location of a tumor suppressor gene (TSG). We analyzed the 2p24 homozygous deletion of a non‐small‐cell lung cancer (NSCLC) cell line, NCI‐H2882, and found that the deletion size was 3.7 Mbp. Since RhoB, which has been suggested to be a candidate TSG, was located in this region, we analyzed RhoB for alterations in NSCLC. Although we found no mutations in 48 cell lines including 20 NSCLCs, a loss of heterozygosity (LOH) analysis in 128 primary NSCLCs showed that 25 of 62 informative samples had LOH at the RhoB locus. Northern blot analysis of 28 cell lines (including 15 NSCLCs) indicated that RhoB expression was downregulated in 27. We analyzed RhoB expression in 112 primary NSCLCs with immunohistochemistry and found no or a weak RhoB expression in 33 (42%) of 78 adenocarcinomas, whereas we found it in 29 (94%) of 31 squamous cell carcinomas. No or a weak expression of RhoB was more frequently observed in poorly‐ or moderately‐differentiated adenocarcinomas than in well‐differentiated ones (p = 0.0014). Furthermore, no or a weak expression of RhoB indicated a tendency to poor patient prognosis. Although hypermethylation was not found at the promoter region, the RhoB expression in NSCLC cell lines was induced by histone deacetylase inhibition, suggesting that RhoB downregulation may be due to histone modification. The present study demonstrates that RhoB expression is frequently downregulated in NSCLCs by multiple mechanisms, suggesting that RhoB is a candidate TSG for NSCLC. © 2006 Wiley‐Liss, Inc.
Selective cyclooxygenase‐2 inhibitor downregulates the paracrine epithelial–mesenchymal interactions of growth in scirrhous gastric carcinoma
Authors: Yashiro Masakazu , Nakazawa Kazunori , Tendo Masashige , Kosaka Kinshi , Shinto Osamu , Hirakawa Kosei
Published: November 2006

The importance of cancer‐mesenchymal interactions in the aggressive behavior of scirrhous gastric cancer is supported by experimental and clinical evidences. We have previously reported that gastric fibroblasts secretion of keratinocyte growth factor (KGF) underline the remarkable proliferation of scirrhous gastric cancer cells. Cyclooxygenase‐2 (COX‐2) is not only expressed in cancer cells, but also in interstitial fibroblasts in gastric carcinoma. To clarify the mechanisms responsible for the antiproliferation effect of COX‐2 inhibitors, effect of COX‐2 inhibitor on the paracrine epithelial–mesenchymal interactions of growth was examined. Scirrhous gastric cancer cell line, OCUM‐2M, gastric fibroblasts, NF‐21, and COX‐2 inhibitor, JTE‐522, were used. Growth‐interaction was examined by calculating the number of cancer cells or by measuring [3H] thymidine incorporation of cancer cells. Effect of JTE‐522 on KGF expression from NF‐21 cells and OCUM‐2M cells was analyzed by ELISA and RT‐PCR. The conditioned medium from gastric fibroblasts significantly stimulated the growth of scirrhous gastric cancer cells. JTE‐522 at the concentrations of 10−5 and 10−6 M significantly decreased the growth‐stimulating activity of gastric fibroblasts. JTE‐522 reduced the expression of KGF mRNA and the production of KGF from gastric fibroblasts. Oral administration of JTE‐522 significantly decreased the size of xenografted tumor coinoculated with OCUM‐2M cells and NF‐21 cells in nude mice. JTE‐522 decreased COX‐2 expression and Ki67 labeling index within the coinoculated tumor. These findings suggested that a selective COX‐2 inhibitor, JTE‐522, downregulates KGF production from gastric fibroblasts, resulting in the inhibition of paracrine epithelial–mesenchymal interactions of proliferation between scirrhous gastric cancer cells and gastric fibroblasts. © 2006 Wiley‐Liss, Inc.
MDM2 SNP309 T>G alone or in combination with the TP53 R72P polymorphism does not appear to influence disease expression and age of diagnosis of colorectal cancer in HNPCC patients
Authors: Talseth Bente A. , Meldrum Cliff , Suchy Janina , Kurzawski Grzegroz , Lubinski Jan , Scott Rodney J.
Published: November 2006

Disease expression in hereditary nonpolyposis colorectal cancer (HNPCC) cannot be readily explained by mutation site in the respective DNA mismatch repair genes associated with this disorder. One explanation is the role of modifying genes that can either promote or prevent disease development on a background of increased risk. Two single nucleotide polymorphisms in MDM2 and TP53 have been shown to be associated with younger ages of disease onset in HNPCC (TP53) and Li‐Fraumeni syndrome (MDM2). In this study 220 HNPCC patients were examined, from Australia and Poland, all characterized at the molecular level to determine the frequency of the MDM2 SNP309 T>G and to assess its influence on disease expression. The results were then pooled with the results of a previous study to assess the combined influence of the MDM2 SNP309 T>G and TP53 SNP R72P. A significant difference was observed between CRC patients and unaffected MMR gene mutation carriers over the age of 45 years (p = 0.01). The unaffected MMR gene mutation carriers over the age of 45 years who carry the G allele have a reduced risk of developing CRC. The results indicate that the MDM2 SNP309, alone or in combination with TP53 R72P, does not influence age of diagnosis of CRC in individuals with HNPCC. In conclusion, the data indicates the G allele of MDM2 SNP309 might have a protective effect on disease development in HNPCC patients and that age of diagnosis of CRC is not associated with MDM2 SNP309 or TP53 R72P either as single SNPs or combined. © 2006 Wiley‐Liss, Inc.
Comprehensive evaluation of genetic variation in the IGF1 gene and risk of prostate cancer
Authors: Johansson Mattias , McKay James D. , Stattin Pär , Canzian Federico , Boillot Catherine , Wiklund Fredrik , Adami Hans‐Olov , Bälter Katarina , Grönberg Henrik , Kaaks Rudolf
Published: November 2006

Insulin‐like growth factor‐I (IGF1) stimulates cell proliferation, decreases apoptosis, and has been implicated in cancer development. Epidemiological studies have shown elevated levels of circulating IGF1 to be associated with increased risk of prostate cancer. To what extent genetic variation in the IGF1 gene is related to prostate cancer risk is largely unknown. We performed a comprehensive haplotype tagging (HT) assessment of single nucleotide polymorphisms (SNPs) representing the common haplotype variation in the IGF1 gene. We genotyped 10 SNPs (9 haplotype tagging SNPs (htSNPs)) within Cancer Prostate in Sweden (CAPS), a case–control study of 2,863 cases and 1,737 controls, in order to investigate if genetic variation in the IGF1 gene is associated with prostate cancer risk. Three haplotype blocks were identified across the IGF1 gene and 9 SNPs were selected as haplotype tagging SNPs. Common haplotypes in the block covering the 3′ region of the IGF1 gene showed significant global association with prostate cancer risk (p = 0.004), with one particular haplotype giving an odds ratio of 1.46 (95% CI = 1.15–1.84, p = 0.002). This haplotype had a prevalence of 5% in the study population. Our results indicate that common variation in the IGF1 gene, particularly in the 3′ region, may affect prostate cancer risk. Further studies on genetic variations in the IGF1 gene in relation to prostate cancer risk as well as to circulating levels of IGF1 are needed to confirm this novel finding. © 2006 Wiley‐Liss, Inc.
Selective cytotoxicity of benzyl isothiocyanate in the proliferating fibroblastoid cells
Authors: Miyoshi Noriyuki , Uchida Koji , Osawa Toshihiko , Nakamura Yoshimasa
Published: November 2006

In the present study, experiments using presynchronization culture cells demonstrated that benzyl ITC (BITC), previously isolated from a tropical papaya fruit extract, induced the cytotoxic effect preferentially in the proliferating human colon CCD‐18Co cells to the quiescent ones. Quiescent CCD‐18Co cells were virtually unaffected by BITC and marginal cytotoxicity was observed at 15 μM. We observed that BITC dramatically induced the p53 phosphorylation and stabilization only in the quiescent (G0/G1 phase‐arrested) cells, but not significantly in the proliferating human colon CCD‐18Co cells when compared with quiescent ones. We also observed ataxia telangiectasia‐mutated (ATM) phosphorylation in the quiescent cells. The BITC‐induced p53 phosphorylation was counteracted by caffeine treatment, implying the involvement of an ATM/ataxia telangiectasia and Rad3‐related kinase signaling pathway. Moreover, downregulation of p53 by a siRNA resulted in the enhancement of susceptibility to undergo apoptosis by BITC. We also showed here that depletion of p53 abrogated G0/G1 arrest accompanied by the declined expression of p21waf1/cip1 and p27kip1 in CCD‐18Co cells. In conclusion, we identified p53 as a potential negative regulator of the apoptosis induction by BITC in the normal colon CCD‐18Co cells through the inhibition of cell‐cycle progression at the G0/G1 phase. © 2006 Wiley‐Liss, Inc.
Association of interleukin‐1 gene polymorphisms with gastric cancer: A meta‐analysis
Authors: Wang Ping , Xia Harry Hua‐Xiang , Zhang Jian‐Ying , Dai Li‐Ping , Xu Xue‐Qin , Wang Kai‐Juan
Published: November 2006

Previous studies on the association between interleukin‐1 (IL‐1) genetic polymorphisms and the risk of gastric cancer have produced conflicting results. The purpose of this study was to examine the association between IL‐1 genotype and gastric caner by systematically reviewing the risk of the original studies. Thirty‐nine studies, which included 6,863 gastric cancer cases and 8,434 controls, met the inclusion criteria and were included in the meta‐analysis. By pooling all the studies identified, the summary odds ratio (OR) of gastric cancer risk associated with IL‐1B–511T, −31C, +3954T and IL‐1RN*2 was 1.26 (95% confidence interval (CI): 1.03–1.55), 1.00 (95% CI: 0.82–1.22), 1.37 (95% CI: 0.94–2.00) and 1.20 (95% CI: 1.01–1.41), respectively. A stratified analysis showed that IL‐1B–511T was associated with an increased risk of gastric cancer (intestinal type) (OR = 1.76, 95% CI: 1.12–2.57). Moreover, IL‐1RN*2 was also associated with an increased risk of gastric cancer among Caucasians (OR = 1.30, 95% CI: 1.09–1.54). In conclusion, IL‐1B–511 and IL‐1RN genetic polymorphisms are associated with an increased risk of developing gastric cancer. © 2006 Wiley‐Liss, Inc.
Genomic analysis of the 8p11‐12 amplicon in familial breast cancer
Authors: Melchor Lorenzo , Garcia Maria J. , Honrado Emiliano , Pole Jessica C.M. , Alvarez Sara , Edwards Paul A.W. , Caldas Carlos , Brenton James D. , Benítez Javier
Published: November 2006

Amplification of 8p11‐12 has been recurrently reported in sporadic breast cancer. These studies define a complex molecular structure with a set of minimal amplified regions, and different putative oncogenes that show a strong correlation between amplification and over‐expression such as ZNF703/FLJ14299, SPFH2/C8orf2, BRF2 and RAB11FIP. However, none of these studies were carried out on familial breast malignancies. We have studied the incidence, molecular features and clinical value of this amplification in familial breast tumors associated with BRCA1, BRCA2 and non‐BRCA1/2 gene mutations. We detected 9 out of 80 familial tumors with this amplicon by chromosomal comparative genomic hybridization. Next, we used a high‐resolution comparative genomic hybridization array covering the 8p11‐12 region to characterize this chromosomal region. This approach allowed us to define 2 cores of common amplification that largely overlap with those reported in sporadic tumors. Our findings confirm the molecular complexity of this chromosomal region and indicate that this genomic event is a common alteration in breast cancer, present not only in sporadic but also in familial tumors. Finally, we found correlation between the 8p11‐12 amplification and proliferation (Ki‐67) and cyclin E expression, which further proves in familial tumors the poor prognosis association previously reported in sporadic breast cancer. © 2006 Wiley‐Liss, Inc.
PPARγ inhibitors reduce tubulin protein levels by a PPARγ, PPARδ and proteasome‐independent mechanism, resulting in cell cycle arrest, apoptosis and reduced metastasis of colorectal carcinoma cells
Authors: Schaefer Katherine L. , Takahashi Hirokazu , Morales Victor M. , Harris Gianni , Barton Susan , Osawa Emi , Nakajima Atsushi , Saubermann Lawrence J.
Published: November 2006

The nuclear transcription factor peroxisome proliferator‐activated receptor‐gamma (PPARγ) has been identified as an important therapeutic target in murine models of colorectal cancer (CRC). To examine whether PPARγ inhibition has therapeutic effects in late‐stage CRC, the effects of PPARγ inhibitors on CRC cell survival were examined in CRC cell lines and a murine CRC model. Low doses (0.1–1 μM) of PPARγ inhibitors (T0070907, GW9662 and BADGE) did not affect cell survival, while higher doses (10–100 μM) of all 3 PPARγ inhibitors caused caspase‐dependent apoptosis in HT‐29, Caco‐2 and LoVo CRC cell lines. Apoptosis was preceded by altered cell morphology, and this alteration was not prevented by caspase inhibition. PPARγ inhibitors also caused dual G and M cell cycle arrest, which was not required for apoptosis or for morphologic alterations. Furthermore, PPARγ inhibitors triggered loss of the microtubule network. Notably, unlike other standard antimicrotubule agents, PPARγ inhibitors caused microtubule loss by regulating tubulin post‐transcriptionally rather than by altering microtubule polymerization or dynamics. Proteasome inhibition by epoxomicin was unable to prevent tubulin loss. siRNA‐mediated reduction of PPARγ and PPARδ proteins did not replicate the effects of PPARγ inhibitors or interfere with the inhibitors' effects on apoptosis, cell cycle or tubulin. PPARγ inhibitors also reduced CRC cell migration and invasion in assays in vitro and reduced both the number and size of metastases in a HT‐29/SCID xenograft metastatic model of CRC. These results suggest that PPARγ inhibitors are a novel potential antimicrotubule therapy for CRC that acts by directly reducing microtubule precursors. © 2006 Wiley‐Liss, Inc.
In vivo evaluation of the role of DNp73α protein in regulating the p53‐dependent apoptotic pathway after treatment with cytotoxic drugs
Authors: Sabatino Maria Antonietta , Previdi Sara , Broggini Massimo
Published: November 2006

The amino terminus truncated p73 isoform, ΔNp73α, shows dominant negative behavior toward TAp73 and wild‐type p53, and has oncogenic potential. By contrast, we recently showed that in HCT116 clones forced expression of ΔNp73α did not increase in vitro cellular resistance to anticancer agents. The purpose of this study was to characterize in vivo models and to investigate the functional interaction between the ΔNp73α isoform and the p53 pathway. Human colon carcinoma HCT116 clones expressing inducible ΔNp73α (HCT116/DN3, HCT116/DN14) and HCT116/8a (transfected with the mock empty vector), transplanted in immunodeficient nude mice, were used to study the antitumor activity of cis‐diammine‐dichloro‐platinum (cDDP) (4 mg/kg, i.v., q7d × 3) and Doxorubicin (DX) (7.5 mg/kg, i.v., q7d × 3), with or without tetracycline‐induced ΔNp73α overexpression. ΔNp73α expression was confirmed by RT‐PCR, immunoblotting and immunohistochemical analysis. ΔNp73α subcellular localization after DX treatment was checked by an immunofluorescence assay. Western blot was used to analyze p53, p21, Bax, Bcl‐2 and p53AIP1 expression. ΔNp73α overexpression did not modify the antitumor activity of either DX or cDDP in xenograft models. DX reduced ΔNp73α protein expression, without affecting its nuclear localization. p53, p21, Bax and p53AIP1 protein expression increased and Bcl‐2 decreased in HCT116 clone derived tumors 24 hr after DX exposure, independently of the presence of ΔNp73α. Overexpression of ΔNp73α does not affect tumor growth in vivo, does not increase the resistance of established tumors to anticancer agents and does not antagonize p53 apoptotic functions. © 2006 Wiley‐Liss, Inc.
Cetuximab induce antibody‐dependent cellular cytotoxicity against EGFR‐expressing esophageal squamous cell carcinoma
Authors: Kawaguchi Yoshihiko , Kono Koji , Mimura Kousaku , Sugai Hidemitsu , Akaike Hidenori , Fujii Hideki
Published: November 2006

To evaluate the possibility of treatment with antiepidermal growth factor receptor (EGFR) mAb, Cetuximab against esophageal squamous cell carcinoma (SCC), we performed detail analysis of the antibody‐dependent cellular cytotoxicity (ADCC) mediated by Cetuximab against esophageal SCC. Esophageal SCC cell lines with various levels of EGFR (n = 8) were evaluated for their Cetuximab‐mediated ADCC by 51Cr‐release assay. As a result, Cetuximab was able to induce ADCC against EGFR‐expressing esophageal SCC and the activities reflected the degree of EGFR expression on the esophageal SCC. The activities of Cetuximab‐mediated ADCC by patients' PBMC were impaired in comparison with those by healthy donors' PBMC. Moreover, the inhibition of transforming growth factor (TGF)‐β could enhance Cetuximab‐mediated ADCC against TGF‐β‐producing SCC. In conclusion, Cetuximab was able to induce ADCC against EGFR‐expressing esophageal SCC. Some modalities aiming at enhancing the Cetuximab‐mediated ADCC may be necessary for successful Cetuximab treatment of patients with esophageal SCC. © 2006 Wiley‐Liss, Inc.
Endogenous adenosine inhibits CNS terminal Ca2+ currents and exocytosis
Authors: Knott Thomas K. , Marrero Héctor G. , Fenton Richard A. , Custer Edward E. , Dobson James G. , Lemos José R.
Published: November 2006

Bursts of action potentials (APs) are crucial for the release of neurotransmitters from dense core granules. This has been most definitively shown for neuropeptide release in the hypothalamic neurohypophysial system (HNS). Why such bursts are necessary, however, is not well understood. Thus far, biophysical characterization of channels involved in depolarization‐secretion coupling cannot completely explain this phenomenon at HNS terminals, so purinergic feedback mechanisms have been proposed. We have previously shown that ATP, acting via P2X receptors, potentiates release from HNS terminals, but that its metabolite adenosine, via A1 receptors acting on transient Ca2+ currents, inhibit neuropeptide secretion. We now show that endogenous adenosine levels are sufficient to cause tonic inhibition of transient Ca2+ currents and of stimulated exocytosis in HNS terminals. Initial non‐detectable adenosine levels in the static bath increased to 2.9 µM after 40 min. These terminals exhibit an inhibition (39%) of their transient inward Ca2+ current in a static bath when compared to a constant perfusion stream. CPT, an A1 adenosine receptor antagonist, greatly reduced this tonic inhibition. An ecto‐ATPase antagonist, ARL‐67156, similarly reduced tonic inhibition, but CPT had no further effect, suggesting that endogenous adenosine is due to breakdown of released ATP. Finally, stimulated capacitance changes were greatly enhanced (600%) by adding CPT to the static bath. Thus, endogenous adenosine functions at terminals in a negative‐feedback mechanism and, therefore, could help terminate peptide release by bursts of APs initiated in HNS cell bodies. This could be a general mechanism for controlling transmitter release in these and other CNS terminals. J. Cell. Physiol. 210: 309–314, 2007. © 2006 Wiley‐Liss, Inc.
Cisplatin upregulates Saccharomyces cerevisiae genes involved in iron homeostasis through activation of the iron insufficiency‐responsive transcription factor Aft1
Authors: Kimura Akiko , Ohashi Kazuaki , Naganuma Akira
Published: November 2006

The response of Saccharomyces cerevisiae to cisplatin was investigated by examining variations in gene expression using cDNA microarrays and confirming the results by reverse transcription polymerase chain reaction (RT‐PCR). The mRNA levels of 14 proteins involved in iron homeostasis were shown to be increased by cisplatin. Interestingly, the expression of all 14 genes is known to be regulated by Aft1, a transcription factor activated in response to iron insufficiency. The promoter of one of these genes, FET3, has been relatively well studied, so we performed a reporter assay using the FET3 promoter and showed that an Aft1 binding site in the promoter region is indispensable for induction of transcription by cisplatin. The active domain of Aft1 necessary for activation of the FET3 promoter by cisplatin is identical to the one required for activation by bathophenanthroline sulfonate, an inhibitor of cellular iron uptake. Furthermore, we found that cisplatin inhibits the uptake of 55Fe(II) into yeast cells. These findings suggest that cisplatin activates Aft1 through the inhibition of iron uptake into the cells, after which the expression of Aft1 target genes involved in iron uptake might be induced. J. Cell. Physiol. 210: 378–384, 2007. © 2006 Wiley‐Liss, Inc.