bluuebunny/crossref_metadata_2025 huggingface dataset:
Derivatives of Coenzyme F430 with a Covalently Attached α‐Axial Ligand. Part II
Authors: Bauer Carsten , Jaun Bernhard
Published: December 2003

Coenzyme F430 pentamethyl ester 2 was partially hydrolyzed to a mixture of the five F430 tetramethyl esters 7–11, which were separated by HPLC and identified by means of a full NMR characterization. The tetramethyl ester with a free COOH group at the side chain at C(3) of F430 was coupled to the N‐terminus of the peptidic spacerligand construct 12 selected and studied as described before. The UV/VIS and NMR spectra in CH2Cl2/3,3,3‐trifluoroethanol 6 : 1 show that the new derivative, the NiII(33‐dehydroxy‐83,122,133,182‐tetra‐O‐methyl‐F430‐33‐yl)‐L‐prolyl‐L‐prolyl‐Nπ‐methyl‐L‐histidine methyl ester (13), is an intramolecular, pentacoordinate, paramagnetic complex. In the same solvent system, the parent 33,83,122,133,182‐penta‐O‐methyl‐F430 (2) is four coordinate and diamagnetic even in the presence of equimolar 1H‐imidazole. Protonation of the axially coordinating histidine residue of 13 gave the diamagnetic tetracoordinate base‐off form, which allowed us to establish the constitution of 13 by NMR.
Pentopyranosyl Oligonucleotide Systems. 9th Communication
Authors: Pitsch Stefan , Wendeborn Sebastian , Krishnamurthy Ramanarayanan , Holzner Armin , Minton Mark , Bolli Martin , Miculca Christian , Windhab Norbert , Micura Ronald , Stanek Michael , Jaun Bernhard , Eschenmoser Albert
Published: December 2003

Pyranosyl‐RNA (‘p‐RNA’ ) is an oligonucleotide system isomeric to natural RNA and composed of the very same building blocks as RNA. Its generational, chemical, and informational properties are deemed to be those of an alternative nucleic acid system that could have been a candidate in Nature's evolutionary choice of the molecular basis of genetic function. We consider the study of the chemistry of p‐RNA as etiologically relevant in the sense that knowledge of its structural, chemical, and informational properties on the chemical level offers both a perspective and reference points for the recognition of specific structural assets of the RNA structure that made it the (supposedly) superior system among possible alternatives and, therefore, the system that became part of biology as we know it today. The paper describes the chemical synthesis of β‐d‐ (and L)‐ribopyranosyl‐(4′→2′)‐oligonucleotide sequences, presents a resume of their structural and chemical properties, and cautiously discusses what we may and may not have learned from the pyranosyl isomer of RNA with respect to the conundrum of RNA's origin.
Nizatidine for the treatment of patients with quetiapine‐induced weight gain
Authors: Atmaca Murad , Kuloglu Murat , Tezcan Ertan , Ustundag Bilal , Kilic Nulufer
Published: January 2004

It has been reported that nizatidine may reduce weight gain in schizophrenic patients on olanzapine treatment. Leptin has been reported to be associated with antipsychotic‐induced weight gain. Thus, the purpose of the study was to evaluate whether nizatidine might be useful for the treatment of quetiapine‐induced weight gain. Among the patients on the quetiapine monotherapy, 47 participated in the study for the two and half months of the open‐label screening period. However, 28 patients who gained considerable weight in this period entered the 8‐week, double‐blind and placebo‐controlled phase. These patients were randomly divided into two groups; quetiapine plus nizatidine (group I) and quetiapine plus placebo (group II) for the 8‐week double‐blind phase. The patients were evaluated at the baseline and at week 8 with respect to the positive and negative syndrome scale, body mass index, weight and serum leptin levels. The mean weight and leptin levels exhibited modest increases in both groups for the open‐label screening period. In the double‐blind period, in group I, a minimal, but not statistically significant, decrease in weight was observed, with a mean of 1.0 ± 0.6 kg. The weight increased in group II. The leptin levels decreased by a mean of 0.6 ± 0.6 ng/ml in group I, and increased by 1.0 ± 0.9 ng/ml in group II. At evaluation at week 8, a trend toward statistical significance in the mean serum leptin levels between groups was detected. The results suggest that nizatidine treatment may stop but not reduce the weight gain and is correlated with leptin levels in patients with schizophrenia on quetiapine treatment. Copyright © 2003 John Wiley & Sons, Ltd.
Effect of glucose on tobacco withdrawal symptoms in recent quitters using bupropion or nicotine replacement
Authors: McRobbie Hayden , Hajek Peter
Published: January 2004

Rationale and objectives

Glucose has been shown to alleviate the desire to smoke in abstaining heavy smokers and to increase 1‐month abstinence rates on its own as well as when combined with nicotine patches. It is not known whether a single dose of glucose can provide additional withdrawal relief in patients who have already abstained for a period of time, and whether it can assist patients using bupropion.

Methods

Seventy‐five volunteers from a smoking cessation clinic who maintained 1 week of validated continuous abstinence were randomized to receive four 3 mg dextrose or placebo (sorbitol) tablets. There were 31 bupropion and 44 NRT users. Measures of desire to smoke and of five withdrawal symptoms were taken before taking the tablets, and then at 5 min intervals for 20 min.

Results

Despite low baseline ratings of withdrawal discomfort, glucose tablets significantly reduced irritability and hunger in bupropion users. Two other effects including reduction in composite withdrawal score approached but did not reach statistical significance. The effects emerged 10–15 min after taking the tablets. No glucose effect was detected in patients using NRT.

Conclusions

A single dose of glucose taken after a week of sustained abstinence may reduce withdrawal discomfort in patients on bupropion. Further research is warranted in combining glucose and bupropion, and in opportunistic use of glucose tablets in tempting situations during a smoking cessation attempt. Copyright © 2004 John Wiley & Sons, Ltd.
Olanzapine treatment of eight adolescent patients with psychosis
Authors: Ercan Eyup Sabri , Kutlu Ayşe , Varan Azmi , Çikoğlu Sibel , Coşkunol Hakan , Bayraktar Erhan
Published: January 2004

Although olanzapine is known as a widely used atypical antipsychotic there have been very few studies about its use in children and adolescents. Eight adolescent patients who were diagnosed as having schizophrenia or schizoaffective disorder, and treated with olanzapine are reported in this case series. The patients were followed‐up for 17.5 weeks in the range 4–26 weeks. According to the CGI improvement assessment at the end of the follow‐up period, three of eight cases were rated as very much or much improved, three as minimally improved and two as not improved. Olanzapine was well tolerated by the adolescents in this case series except for weight gain. Our results suggest that olanzapine may be an effective antipsychotic for some psychotic adolescents and during olanzapine trials weight gain. Should be monitored. Copyright © 2004 John Wiley & Sons, Ltd.
In vitro microsomal metabolism of imipramine under conditions mimicking the in vivo steady‐state situation
Authors: Linnet Kristian
Published: January 2004

A steady‐state model is presented of the metabolism of imipramine. In this model, the N‐demethylation and 2‐hydroxylation rates of imipramine were measured with and without the presence of metabolites by using a tritium isotope of imipramine. At an imipramine concentration of 5 μmol/l, desipramine, in the concentration ratio 1:2, decreased the total metabolic rate by 70%. The 2‐hydroxylation pathway was mainly inhibited, thereby increasing the N‐demethylation pathway from 25% to 62% in the presence of desipramine. The additional presence of 2‐hydroxy‐imipramine did not change this situation. A study on the relative influence of CYP1A2 and 3A4 only revealed minor changes in the presence of desipramine. In conclusion, the presence of metabolites in metabolism studies undertaken in vitro may reflect the changes from the single‐ to the multiple‐dose situation observed clinically and therefore constitute a better model for the clinical application of a drug. Copyright © 2004 John Wiley & Sons, Ltd.
The treatment of depression with different formulations of venlafaxine: a comparative analysis
Authors: Olver James S. , Burrows Graham D. , Norman Trevor R.
Published: January 2004

Venlafaxine is the first of a group of antidepressants that show dual reuptake inhibition of serotonin and noradrenaline (SNRIs). Originally marketed in an immediate release (IR) formulation a microencapsulated, extended release (XR) formulation is now available. Significant differences exist between these two formulations with respect to pharmacokinetic parameters which have an impact on clinical use. The XR has lower maximum plasma concentrations (Cmax) and achieves these at a later time (higher Tmax). The longer apparent elimination half‐life of the drug after single XR doses suggests that it is suitable for once daily dosing compared with the twice daily dosing regimen required by the IR formulation. With respect to antidepressant efficacy the XR formulation is equivalent to other marketed antidepressants and to the IR formulation. Consistent with its pharmacokinetic properties the use of the XR formulation is associated with less nausea and dizziness at the initiation of therapy. While in clinical usage XR might be expected to increase compliance with medication and to reduce discontinuation syndromes there are few comparative studies for which this has been evaluated. The XR formulation of venlafaxine is no worse than the IR form with respect to tolerability and offers some benefits to patients in terms of ease of use. On the other hand there does not appear to be any increase in the efficacy of the active agent. Copyright © 2004 John Wiley & Sons, Ltd.
Double‐blind comparative study of the action of repeated administration of milnacipran versus placebo on cognitive functions in healthy volunteers
Authors: Poirier M. F. , Galinowski A. , Amado I. , Longevialle R. , Bourdel M. C. , Tournoux A. , Serre C. , Loo H.
Published: January 2004

Objective

The principal objective was to compare the effects of milnacipran, an antidepressant characterized by a dual‐action on serotonin and noradrenaline reuptake, with placebo on memory, attention and psychomotor performance in healthy volunteers. The secondary objective was to evaluate the effects of milnacipran on mood, anxiety and vigilance in these subjects.

Methods

In a double‐blind crossover randomized trial, milnacipran (50 mg b.d.) or placebo was administered during two periods of 7 days separated by a washout period of 7 days. Memory tests (recall of words, images and coloured bars), tests to evaluate attention and vigilance (squares test, critical flicker fusion test and choice reaction time test) and visual analogue scales for affect and sleep were used.

Results

There were no significant differences between milnacipran and placebo groups with respect to the psychomotor functions tested. No differences were observed in the Norris scales for vigilance, anxiety or satisfaction or in the sleep questionnaire (sleep latency, sleep quality and waking).

Conclusion

Milnacipran, administered at 100 mg per day for 7 days to healthy volunteers, had no effects on cognitive functions. Copyright © 2004 John Wiley & Sons, Ltd.
P16INK4a as an adjunct marker in liquid‐based cervical cytology
Authors: Sahebali Shaira , Depuydt Christophe E. , Segers Kurt , Moeneclaey Liliane M. , Vereecken Annie J. , Van Marck Eric , Bogers Johannes J.
Published: January 2004

Cytological screening for cervical cancer is hampered by high false negative rates. Inter‐observer reproducibility needs optimizing. The potential of p16INK4a as a biomarker for cervical lesions was examined in a study of liquid‐based cytology (LBC), HPV DNA testing by MY09/MY11 consensus PCR and type‐specific PCRs and p16INK4a immunocytochemistry on a series of 291 patients selected from routine screening. Comparison of the number of p16INK4a immunoreactive cells/1,000 cells exhibited a significantly higher mean count in HSIL (8.80 ± 1.13) than other cytological groups. The mean count of LSIL (1.09 ± 0.18) was significantly higher than that of the negative group (0.82 ± 0.40). ASC‐H and HSIL combined showed a significantly higher mean count (6.46 ± 1.17) than negative, ASC, ASC‐US and LSIL. The mean count of immunoreactive cells/1,000 cells was significantly higher in HPV16 positive samples (3.22 ± 0.72) than in samples containing infections with types of unknown malignant potential (0.83 ± 0.26) or HPV negative samples (1.17 ± 0.41). The mean count in infections with other high‐risk HPV types (2.55 ± 0.52) was significantly higher than that in HPV negative samples. Receiver‐operating characteristic curves yielded a test accuracy (area under curve) of 0.76, 0.79, 0.88 and 0.95 for ASCUS, LSIL, ASC‐H/HSIL and HSIL, respectively. Thresholds for 95% sensitivity were at 0.005, 0.007, 0.098 and 0.445 immunopositive cells/1,000 cells for ASCUS, LSIL, ASC‐H/HSIL and HSIL, respectively. The 95% specificity threshold for the detection of HSIL was at 1.87 immunopositive cells/1,000 cells. P16INK4a immunocytochemistry can be used as an adjunct to LBC in cervical screening, because it has a good diagnostic accuracy to discriminate HSIL and ASC‐H from other lesions. It could be used as a surrogate marker of high‐risk HPV infections. © 2003 Wiley‐Liss, Inc.
Effect of vitamin D analog (1α hydroxy D5) immunoconjugated to Her‐2 antibody on breast cancer
Authors: Punj Vasil , Graves Jewell M. , Mehta Rajeshwari R.
Published: January 2004

We previously showed that a new vitamin D analog, 1α(OH)D5 (D5), induced differentiation and inhibited the growth of breast cancer cells. In this report, we examined whether D5 specifically delivered to breast cancer cells could have any therapeutic effect. D5 was linked to Her‐2 antibody using sulfosuccinimidyl 6‐4 azido nitrophenylamido hexanode (SANPAH) as a linker. The Her‐2 antibody selected in our study had no significant effect on the in vitro or in vivo growth of breast cancer cells; however, it had cell‐differentiating action. In vitro, D5‐Her‐2 antibody conjugate (IMC) showed the ability to specifically bind to Her‐2‐expressing cells, to compete with Her‐2 antibody for surface receptor and to cause internalization. IMC (equivalent to 5 μg Her‐2 antibody given intraperitoneally once weekly for 6 weeks) significantly inhibited the growth of BT‐474 cells transplanted into athymic mice. The in vivo growth‐inhibitory effect of IMC treatment was similar to that observed in animals receiving D5 continuously as a dietary supplement. These results show that the targeted delivery of D5 by immunoconjugation to cell surface receptor antibodies may be of potential therapeutic value for the treatment of Her‐2 positive breast cancer. © 2003 Wiley‐Liss, Inc.
Analysis of the level of mRNA expression of the membrane regulators of complement, CD59, CD55 and CD46, in breast cancer
Authors: Rushmere Neil K. , Knowlden Janice M. , Gee Julia M.W. , Harper Maureen E. , Robertson John F. , Morgan B. Paul , Nicholson Robert I.
Published: January 2004

We have examined the relative mRNA expression of the complement (C) regulatory proteins CD59, CD55 and CD46 in RNA isolated from 50 primary breast cancer specimens using a semiquantitative RT‐PCR approach. Having normalized the mRNA expression levels of the C regulators relative to actin, we subsequently correlated their expression with estrogen receptor (ER) and various clinical, pathologic and biochemical features of the disease. CD59 and CD46 were detected in all clinical biopsies, while CD55 mRNA was detected in the majority of samples. The comparative levels of expression between the 3 regulators analyzed, using Spearman rank correlation test, revealed a significant association (p = 0.01; r = 0.36) between CD46 and CD59. CD46 exhibited the most striking pattern of association, with increased levels of expression being associated with ER‐positive samples and lower levels of expression associated with a loss of differentiation and epidermal growth factor receptor positivity. Application of Spearman rank correlation test revealed CD46 expression was significantly associated with expression of ER at the level of protein (p = 0.031; r = 0.31) and mRNA (p < 0.001; r = 0.52). CD46 expression also correlated with insulin‐like growth factor receptor‐positive samples using Spearman rank correlation test (p = 0.016; r = 0.34), but negatively associated with tumor samples either exhibiting histologic grade 3 when compared to grades 1 or 2 or displaying elevated levels of inflammatory cell infiltrate. Immunohistochemical analysis of a limited series (n = 8) of paraffin‐embedded breast cancers indicated that the level of CD46 protein expression directly associates with that of the mRNA and, where prominent, is localized in the tumor epithelial cell population, including at the plasma membrane. These data provide new information on expression of these important regulators in breast cancer and suggest that CD46 should be evaluated as a novel prognostic indicator. © 2003 Wiley‐Liss, Inc.
Expression of sperm protein 17 (Sp17) in ovarian cancer
Authors: Straughn J. Michael , Shaw Denise R. , Guerrero Antonio , Bhoola Snehal M. , Racelis Alicia , Wang Zhiqing , Chiriva‐Internati Maurizio , Grizzle William E. , Alvarez Ronald D. , Lim Seah H. , Strong Theresa V.
Published: January 2004

Sperm protein 17 (Sp17) is an antigenic protein highly expressed in spermatozoa. Sp17 expression was demonstrated recently in multiple myeloma, suggesting that it may be a novel cancer‐testis antigen. Expression of Sp17 mRNA and protein was examined in human ovarian tumors. Sp17 mRNA was evaluated by reverse transcription‐polymerase chain reaction (RT‐PCR) and Northern blot analysis of RNA derived from epithelial ovarian tumors and normal tissues. RT‐PCR analysis detected Sp17 transcripts in 15 of 18 (83%) primary ovarian tumors. The transcript was not detected in RNA derived from normal uterus or cervix, whereas weak expression was noted in some normal ovarian tissue samples. Northern blot analysis showed no detectable Sp17 mRNA expression in normal tissues, including normal ovary, but showed Sp17 expression in 17 of 25 ovarian tumors (68%). To evaluate protein expression, mouse monoclonal antibodies were produced against recombinant Sp17 protein and used in Western blot and immunohistochemical analyses of normal reproductive tissue and primary ovarian tumor samples. Sp17 protein was detected by Western blot analysis in normal spermatozoa and in 8 of 19 ovarian tumor samples. Immunohistochemical studies showed Sp17 expression in spermatozoa, ciliated cells of the female reproductive tract, and most ovarian tumors evaluated. Tumors showed a predominately nuclear localization of Sp17 expression, with some cytoplasmic staining. These results demonstrate that Sp17, a protein with restricted expression in somatic tissues, is expressed in ovarian tumors. Because Sp17 is immunogenic, it may represent a novel target for immunotherapeutic interventions for ovarian cancer patients. © 2003 Wiley‐Liss, Inc.
Screen‐detected prostate cancer and the insulin‐like growth factor axis: Results of a population‐based case‐control study
Authors: Oliver Steven E. , Gunnell David , Donovan Jenny , Peters Tim J. , Persad Raj , Gillatt David , Pearce Anya , Neal David E. , Hamdy Freddie C. , Holly Jeff
Published: January 2004

Higher circulating levels of IGF‐I have been associated with increased risk of prostate and some other cancers. Most research on prostate cancer has been based on men with symptoms or identified following treatment of benign disease. However, increasing numbers of cancer cases are now detected in asymptomatic men following prostate‐specific antigen (PSA) tests. We therefore used a population‐based case‐finding exercise using the PSA test to examine whether associations between the IGF axis and cancer risk were apparent in this population. A matched case‐control study was conducted among 7,383 men (50–70 years) receiving a PSA test as part of a case‐finding exercise. Assays of IGF‐I, IGF‐II, IGFBP‐2 and IGFBP‐3 were performed on cases and 2 controls matched on age, recruitment center and calendar time. Analyses were based on 176 cases and 324 matched controls. The risk of prostate cancer increased across quartiles of IGF‐I (highest vs. lowest quartile, OR = 2.34; 95% CI = 1.26–4.34; ptrend = 0.02) and IGF‐II (OR = 1.78; 95% CI = 0.94–3.15; ptrend = 0.09). Controlling for smoking history and IGFBP‐3 strengthened associations with cancer for both IGF‐I (OR = 3.00; 95% CI = 1.50–6.01; ptrend 0.005) and IGF‐II (OR = 2.02; 95% CI = 1.07–3.84; ptrend = 0.04) Associations between the IGFs and cancer risk were stronger for advanced cases. Our findings suggest that both IGF‐I and IGF‐II are associated with an increased risk of screen‐detected prostate cancer. © 2003 Wiley‐Liss, Inc.
HLA‐B35‐restricted immune responses against survivin in cancer patients
Authors: Reker Sine , Becker Jürgen C. , Svane Inge Marie , Ralfkiaer Elisabeth , Straten Per thor , Andersen Mads Hald
Published: January 2004

Two HLA‐A2 restricted epitopes have recently been identified from the broadly expressed tumor antigen survivin, and several vaccination trials in cancer patients based on these survivin‐derived peptides have been initiated. Consequently, there is a crucial need for the identification of survivin epitopes restricted to other HLA‐molecules in order to extend the proportion of patients that can enter these ongoing clinical trials. In the present study, we characterized 2 survivin‐derived epitopes, which are restricted to HLA‐B35. Specific T‐cell reactivity against these survivin‐derived epitopes was found in the peripheral blood from patients with different B‐cell malignancies and melanoma. Substitution of the C‐terminal anchor residue of the survivin‐derived peptides improved the recognition by tumor‐infiltrating lymphocytes from melanoma patients. Furthermore, we demonstrated spontaneous cytotoxic T‐cell responses to survivin in a primary melanoma lesion. The characterization of these epitopes allows more patients can be included in the ongoing peptide‐based survivin vaccination trials against cancer. © 2003 Wiley‐Liss, Inc.
Soft‐tissue sarcoma and dioxin: A case‐control study
Authors: Tuomisto Jouni T. , Pekkanen Juha , Kiviranta Hannu , Tukiainen Erkki , Vartiainen Terttu , Tuomisto Jouko
Published: January 2004

Soft‐tissue sarcoma has been proposed to be a candidate for a dioxin‐induced cancer. However, previous epidemiologic studies have suffered from poor exposure data and mixed exposures. We studied the association between sarcoma risk and individually estimated dioxin exposure in a general population exposed to relatively low levels of dioxin via food. A multicenter prospective case‐control study was conducted in 4 university hospitals and 12 other hospitals in southern Finland. Participants included 110 patients with soft‐tissue sarcoma (cases) and 227 area‐ and age‐matched controls. Controls were patients operated for appendicitis. Individual dioxin concentrations were analyzed from subcutaneous fat samples by gas chromatography‐mass spectrometry. The average (range) dioxin concentration was 33.4 (4.4–145.5) ng/kg (toxic equivalencies in fat according to WHO). No increased risk associated with increased dioxin concentration was observed. In contrast, the highest risk of sarcoma was found at low levels of dioxin. Odds ratios for different quintiles as compared with the lowest quintile of dioxins (median, 11.5 ng/kg) varied from 0.43 (95% CI = 0.18–1.05) to 0.65 (95% CI = 0.22–1.95). The result was little affected by studied confounders and the findings were similar for different sarcoma subtypes, age groups and study areas. The results imply that dioxin does not increase the risk of soft‐tissue sarcoma at the present population levels. © 2003 Wiley‐Liss, Inc.
Fine mapping of chromosome 22q tumor suppressor gene candidate regions in astrocytoma
Authors: Hartmann Christian , Nümann Astrid , Mueller Wolf , Holtkamp Nikola , Simon Matthias , von Deimling Andreas
Published: January 2004

Astrocytomas and glioblastomas are the most frequent primary brain tumors in adults. Mutations and altered expression of multiple genes have been found to contribute to the genesis of these tumors. However, many factors in the genesis of astrocytic gliomas are not resolved yet. The frequent losses on several chromosomes indicate the role of still unidentified tumor suppressor genes. Loss of heterozygosity (LOH) on 22q has been described in up to 30% of astrocytic tumors and may be associated with progression to anaplasia. In a first step, information from the nearly finished physical sequence of chromosome 22 were used to map LOH data from 22q deletion studies on different tumor entities to identify potential tumor suppressor gene candidate regions. Next, a series of 153 astrocytic gliomas was examined with 11 polymorphic markers spanning these regions. Forty‐nine (32%) astrocytic gliomas exhibited LOH on 22q, 17 (35%) of which lost heterozygosity for all markers and 32 (65%) of which carried interstitial or partial deletions. Two regions were identified on the physical DNA sequence. The centromeric region spans 3 Mb and the telomeric region 2.7 Mb. The reduced size of these regions now allows direct analysis of all genes included. We already performed mutation analysis on 4 candidate genes from these regions (MYO18B, DJ1042K10.2, MKL1 and EP300), but did not find any mutations in astrocytic tumors. © 2003 Wiley‐Liss, Inc.
Nadir prostate‐specific antigen best predicts the progression to androgen‐independent prostate cancer
Authors: Morote Juan , Trilla Enrique , Esquena Salvador , Abascal José María , Reventos Jaume
Published: January 2004

The objective of our study was to analyze the value of prostate‐specific antigen (PSA) levels before and after androgen suppression to predict the time to androgen‐independent progression (AIP) in patients with advanced and metastatic prostate cancer. A series of 283 prostate cancer patients under androgen suppression as a single treatment was studied. The disease was locally advanced in 98 patients and metastatic in the remainder 185. AIP was defined after 2 consecutive increases of serum PSA after the nadir value. The mean follow‐up before AIP was 29.2 months (3–198). AIP was detected in 205 patients (72.4%). In 152 patients (74.1%), the event was detected within 24 months, while in 53 patients (25.9%), it was observed beyond 24 months. The multivariate analysis showed that the nadir PSA and the time to reach the nadir PSA were the most significant predictors of the time to AIP. The odds ratio of having a biochemical response greater than 24 months was 20 times higher in patients that achieved an undetectable PSA level of 0.2 ng/mL or less. Moreover in those patients whose nadir PSA reached beyond 12 months after androgen suppression the odds ratio was 18 times higher. These results show that the ability to achieve an undetectable nadir PSA and the time to reach it are the most significant predictors of the time to AIP in patients with locally advanced and metastatic prostate cancer under androgen suppression as a single therapy. © 2003 Wiley‐Liss, Inc.
Enhanced radiosensitization and chemosensitization in NF‐κB‐suppressed human oral cancer cells via the inhibition of γ‐irradiation‐ and 5‐FU‐induced production of IL‐6 and IL‐8
Authors: Tamatani Tetsuya , Azuma Masayuki , Ashida Yuki , Motegi Katsumi , Takashima Rina , Harada Koji , Kawaguchi Shin‐ichi , Sato Mitsunobu
Published: January 2004

We examined the mechanisms underlying the enhancement of radiosensitivity and chemosensitivity to γ‐irradiation (IR) and 5‐Fluorouracil (5‐FU) in human oral carcinoma cells (B88) in which NF‐κB activity was constitutively suppressed. Three super‐repressor form of IκBα cDNA‐transfected cell (B88mI) clones and 1 empty vector‐transfected cell clone (B88neo) have been established. We found that the tumor‐forming ability in nude mice of B88mI clones was significantly lower than that of B88 or B88neo. This suppressed ability in tumorigenicity was attributed to the down‐regulation of the expression of interleukin (IL)‐1α, IL‐6, IL‐8, vascular endothelial growth factor (VEGF) and matrix metalloproteinase (MMP)‐9 in B88mI cell clones as compared to that in B88 or B88neo. IR and 5‐FU induced a much greater degree of apoptosis, as evidenced by flow cytometry analysis and annexin V staining, in B88mI cell clones than in B88 or B88neo. When tumor‐bearing nude mice were treated with IR or 5‐FU, the suppression of tumor growth was significantly augmented in B88mI cell clones as compared to that in B88 or B88neo. ELISA analysis indicated that although a remarkable increase in production of IL‐6 and IL‐8 was observed in B88 and B88neo after in vitro exposure to IR or treatment with 5‐FU, radiotherapy and chemotherapy‐induced production of these cytokines was significantly suppressed in B88mI cell clones. These findings suggest that production of angiogenic factors and growth factors in response to radiotherapy and chemotherapy is a principal mechanism of inducible radioresistance and chemoresistance in human oral cancers, and establish the inhibition of NF‐κB as a rational approach to improve conventional radiotherapy and chemotherapy outcomes. © 2003 Wiley‐Liss, Inc.
Heterozygote deficiency in thymidylate synthase enhancer region polymorphism genotype distribution in Hungarian colorectal cancer patients
Authors: Adleff Vilmos , Hitre Erika , Köves István , Orosz Zsolt , Hajnal Anna , Kralovánszky Judit
Published: January 2004

Thymidylate synthase (TS) gene polymorphisms are important as prognostic factors in cancer chemotherapy, but recent results describe that the TS enhancer region (TSER) polymorphic genotypes may also modulate risk for malignancies. Two functionally important and ethnically diverse polymorphisms are present on the TS transcript, TSER, a repeat polymorphism (2 or 3 repeats; 2R, 3R) affecting TS expression, and a 6 bp ins/del polymorphism on the 3′ UTR (position TS1494, del6 or ins6), which may influence mRNA stability. Hungarian population has one of the highest colorectal cancer (CRC) mortality rates in Europe, and several elevated dietary risk factors affect a large part of the population. In our study (99 primary CRC cases), population analysis of the patient group genotype frequencies revealed a departure from the Hardy‐Weinberg equilibrium and significant heterozygote deficiency (p < 0.05) at the TSER locus. Despite the strong linkage between the 2 polymorphic loci, case TS1494del genotype frequencies were normally distributed, as well as the genotype frequencies of the healthy control population (n = 102), at both loci. Case‐control comparison demonstrated a lower relative risk of TSER heterozygotes (OR = 0.47; CI = 0.27–0.83; p = 0.008) and a possible higher prevalence of the 3R3R&ins6/del6 in the CRC group. The observation that heterozygotes are those less susceptible for CRC in the Hungarian population may support the possibility of 2 different pathways in which TS may play a role in colorectal carcinogenesis, probably nutrient (or folate)‐dependently. The lack of similar genotype effect seen with TS1494del polymorphism and the increased presence of one genotype combination (3R3R&ins6/del6) in the patient group suggest a possible TS haplotype effect influencing CRC risk. © 2003 Wiley‐Liss, Inc.
Guanine nucleotide depletion triggers cell cycle arrest and apoptosis in human neuroblastoma cell lines
Authors: Messina Elisa , Gazzaniga Paola , Micheli Vanna , Guaglianone Maria Rosaria , Barbato Silvia , Morrone Stefania , Frati Luigi , Aglianò Anna Maria , Giacomello Alessandro
Published: January 2004

Mycophenolic acid (MPA) specifically inhibits inosine‐5′‐monophosphate dehydrogenase, the first committed step toward GMP biosynthesis. In its morpholinoethyl ester pro‐drug form it is one of the most promising immunosuppressive drugs recently developed. The aim of the present study was to investigate the in vitro effects of MPA, at concentrations readily attainable during immunosuppressive therapy, on 3 human neuroblastoma cell lines (LAN5, SHEP and IMR32). Mycophenolic acid (0.1–10 μM) caused a decrease of intracellular levels of guanine nucleotides, a G1 arrest and a time‐ and dose‐dependent death by apoptosis. These effects, associated with an up‐regulation of p53, p21 and bax, a shuttling of p53 protein into the nucleus and a down‐regulation of bcl‐2, survivin and p27 protein, were reversed by the simultaneous addition of guanine or guanosine and were more evident using nondialysed serum containing hypoxanthine. These results suggest that in neuroblastoma cell lines clinically attainable concentrations of mycophenolic acid deplete guanine nucleotide pools triggering G1 arrest and apoptosis through p53‐mediated pathways, indicating a potential role of its morpholinoethyl ester pro‐drug in the management of patients with neuroectodermal tumors. © 2003 Wiley‐Liss, Inc.
Human papillomavirus (HPV) type 16‐specific CD8+ T cell responses in women with high grade vulvar intraepithelial neoplasia
Authors: Todd Richard W. , Roberts Sally , Mann Christopher H. , Luesley David M. , Gallimore Phillip H. , Steele Jane C.
Published: January 2004

Human papillomavirus (HPV)‐associated vulvar intraepithelial neoplasia (VIN) has serious sequelae for the sufferer. Current treatments are associated with poor response and high relapse rates. The development of HPV‐specific T cell immunotherapies offers a new approach to treatment. This will require a detailed understanding of the spectrum of T cell responses induced by HPV antigens, and how effectively viral antigens can be accessed by the immune system. We have investigated the frequency and spectrum of HPV16‐specific CD8+ T cell responses to three HPV16 antigens in 9 women with high grade VIN (VIN3). CD4‐depleted populations of responder cells were screened against overlapping 30–35mer peptides covering the sequences of HPV16 E6, E7 and E4 using ELISPOT assays of IFN‐γ release. We demonstrated CD8+ T cell reactivity to one or more of the proteins in 6 of 9 patient samples. All 6 of these responders recognised peptides covering the E7 protein, 3 of 9 women responded to E6 peptides, but no reactivity was seen to E4. Our results suggest that HPV16‐specific cytotoxic T cells (CTLs) are relatively common in women with persistent VIN3. The HPV‐specific CTL response, however, seems to be ineffective. There is some evidence that there are problems associated with the processing and presentation of HPV antigens by the infected vulvar epithelium. It will be crucial to address this in the design of any T cell based therapy for HPV‐associated VIN and vulval cancer. © 2003 Wiley‐Liss, Inc.
Significance of COX‐2 expression in human renal cell carcinoma cell lines
Authors: Chen Qinzhong , Shinohara Nobuo , Abe Takashige , Watanabe Takafumi , Nonomura Katsuya , Koyanagi Tomohiko
Published: January 2004

Accumulating evidences indicate that cyclooxygenase (COX)‐2 plays an important role in tumorigenesis in many human cancers. Yet the relationship between COX‐2 and human renal cell carcinoma (RCC) remains unclear. The aim of our study was to evaluate COX‐2 expression in human RCC cell lines and its role in tumorigenesis of human RCC. Among the human RCC cell lines (SMKT‐R4, OS‐RC‐2, ACHN) and normal renal cell line RPTEC, COX‐2 overexpression was found in OS‐RC‐2 cells both at mRNA and protein levels. COX‐2 sense‐ and antisense‐orientated vectors were constructed and transferred into RCC cells. Significant suppression of cellular proliferation was demonstrated in OS‐RC‐2 antisense transfectants, whereas promotion was found in SMKT‐R4 sense transfectants by colony‐forming assay despite the observation that COX‐2 specific inhibitor NS398 exhibited similar IC50 among RCC cell lines by MTT assay. In comparison with parent cells and sense transfectants, significant suppression of COX‐2 expression and PGE2 production and increase in butyrate‐induced apoptosis were observed in OS‐RC‐2 antisense transfectants by Western blot, ELISA assay and FACS analysis, respectively. Furthermore, tumor growth and angiogenesis of OS‐RC‐2 antisense transfectants in nude mice was significantly suppressed and the survival time of these mice was significantly prolonged. Our study demonstrates that COX‐2 is overexpressed in OS‐RC‐2 RCC cell line and plays an important role in tumorigenesis of the cells in vivo, which implies that COX‐2 may be a therapeutic target for COX‐2‐expressing RCC, and that suppression of COX‐2 expression by antisense‐based strategy may have potential utility in treatment of COX‐2‐expressing RCC. © 2003 Wiley‐Liss, Inc.
Differential gene‐expression profiling in the leukemia cell lines derived from indolent and aggressive phases of CD56+ T‐cell large granular lymphocyte leukemia
Authors: Daibata Masanori , Matsuo Yoshinobu , Machida Hisanori , Taguchi Takahiro , Ohtsuki Yuji , Taguchi Hirokuni
Published: January 2004

As a rule, T cell large granular lymphocyte (T‐LGL) leukemia runs a chronic clinical course without need for therapy. Some cases, however, progress to an aggressive disease after the indolent clinical stage. The transformation mechanism into a high‐grade malignancy has not been well studied. We have established 2 leukemia cell lines, MOTN‐1 and PLT‐2, derived from the same clone of CD56+ T‐LGL leukemia in chronic and aggressive phases, respectively. The paired availability of such cell lines is valuable in biologic and genetic investigation of T‐LGL leukemia. We used a microarray containing 406 cDNAs to elucidate alterations of gene expression between the 2 cell lines. We found a number of genes that were differentially expressed: 13 genes with increased expression and 3 genes with reduced expression in PLT‐2 cells as compared to MOTN‐1 cells. Increased expression of the dek, rac, Op18, CD6, CD58, CD106, Id2, ATF4, IRF5, ELL2 and D6 genes, and reduced expression of the GzmA and GzmK genes were confirmed by real‐time quantitative reverse transcription‐PCR, whose results paralleled the microarray data. These upregulated genes encode oncoproteins, cell surface antigens including molecules related to T cell proliferation, transcription factors, and a chemokine receptor. The two downregulated genes encode granzymes that play an important role for induction of cell death. These findings suggest that there is differential gene expression in different clinical phases of T‐LGL leukemia and these differentially expressed genes would be potential targets for further studies to identify the genes involved in the transformation process of T‐LGL leukemia. © 2003 Wiley‐Liss, Inc.
Increased expression of c‐Ski as a co‐repressor in transforming growth factor‐β signaling correlates with progression of esophageal squamous cell carcinoma
Authors: Fukuchi Minoru , Nakajima Masanobu , Fukai Yasuyuki , Miyazaki Tatsuya , Masuda Norihiro , Sohda Makoto , Manda Ryokuhei , Tsukada Katsuhiko , Kato Hiroyuki , Kuwano Hiroyuki
Published: January 2004

Transforming growth factor‐β (TGF‐β) regulates cell growth inhibition, and inactivation of the TGF‐β signaling pathway contributes to tumor development. In our previous study, altered expression of TGF‐β, TGF‐β‐specific receptors and Smad4 was shown to correlate with tumor progression in esophageal squamous cell carcinoma (SCC). These components, however, were maintained normally in some patients with esophageal SCC. In our study, the mechanism by which aggressive esophageal SCC maintains these components was investigated, with particular emphasis on the participation of c‐Ski and SnoN as transcriptional co‐repressors in TGF‐β signaling. Immunohistochemistry for c‐Ski and SnoN was carried out on surgical specimens obtained from 80 patients with esophageal SCC. The expression of c‐Ski and SnoN was also studied in 6 established cell lines derived from esophageal SCC and compared to an immortalized human esophageal cell line by Western blotting. High levels of expression of c‐Ski, detected immunohistologically, were found to correlate with depth of invasion (p = 0.0080) and pathologic stage (p = 0.0447). There was, however, no significant correlation between expression of SnoN and clinicopathologic characteristics. A significant correlation between c‐Ski and TGF‐β expression was observed. Moreover, in patients with TGF‐β negative expression, the survival rates of patients with c‐Ski positive expression were significantly lower than those of patients with c‐Ski negative expression (p = 0.0486). c‐Ski was expressed at a high level in 5 of 6 cell lines derived from esophageal SCC compared to immortalized esophageal keratinocytes. Furthermore, the cyclin‐dependent kinase (CDK) inhibitor, p21 that was up‐regulated by TGF‐β signaling was expressed at a low level in the 5 cell lines. The expression of c‐Ski protein as a transcriptional co‐repressor in TGF‐β signaling seems to be correlated with tumor progression of esophageal SCC. © 2003 Wiley‐Liss, Inc.
Frequent hypermethylation of 5′ flanking region of TIMP‐2 gene in cervical cancer
Authors: Ivanova Tatyana , Vinokurova Svetlana , Petrenko Anatolii , Eshilev Ernest , Solovyova Nina , Kisseljov Fjodor , Kisseljova Natalia
Published: January 2004

Tissue inhibitor of metalloproteinase‐2 (TIMP‐2) is an endogenous inhibitor of matrix metalloproteinases (MMPs). This multifunctional protein regulates activities of MMPs and possesses growth promoting effect in cell culture, anti‐tumoral, anti‐apoptotic and anti‐angiogenic effects in animal model systems in vivo. It has been shown that this gene is downregulated in cervical carcinomas. The mechanism of inhibition of TIMP‐2 expression remains obscure. We have examined whether aberrant DNA methylation of the 5′CpG island of the TIMP‐2 gene is involved in its inhibition during cervical carcinogenesis. Bisulfite‐modified DNA sequencing and MSP assay showed aberrant methylation of TIMP‐2 5′‐CpG island in 17 of 36 (47%) invasive cervical carcinomas and in 2 of 3 cervical cancer cell lines. TIMP‐2 gene was mostly unmethylated in the morphologically normal tissues adjacent to the tumors, whereas methylated alleles of this gene were found in 4 samples. Each tumor and each cell line DNA was characterized by unique methylation pattern, however a discrete region of TIMP‐2 CpG island upstream to the transcription start site was densely methylated in all hypermethylated DNA samples examined. The expression of TIMP‐2 mRNA can be restored in the cell lines, in which this discrete region of TIMP‐2 CpG island is methylated, by treatment with demethylating agents, 5‐azacytidine and 5‐aza‐2′‐deoxycytidine. Our data suggest that the aberrant methylation of TIMP‐2 favors the development of primary cervical tumors. We describe for the first time the aberrant hypermethylation of TIMP‐2 gene in human cancer. © 2003 Wiley‐Liss, Inc.
Inducible Hsp70 as target of anticancer immunotherapy: Identification of HLA‐A*0201‐restricted epitopes
Authors: Faure Olivier , Graff‐Dubois Stéphanie , Bretaudeau Laurent , Derré Laurent , Gross David‐Alexandre , Alves Pedro M. S. , Cornet Sébastien , Duffour Marie‐Thérèse , Chouaib Salem , Miconnet Isabelle , Grégoire Marc , Jotereau Francine , Lemonnier François A. , Abastado Jean‐Pierre , Kosmatopoulos Kostas
Published: January 2004

The design of a broad application tumor vaccine requires the identification of tumor antigens expressed in a majority of tumors of various origins. We questioned whether the major stress‐inducible heat shock protein Hsp70 (also known as Hsp72), a protein frequently overexpressed in human tumors of various histological origins, but not in most physiological normal tissues, constitutes a tumor antigen. We selected the p391 and p393 peptides from the sequence of the human inducible Hsp70 that had a high affinity for HLA‐A*0201. These peptides were able to trigger a CTL response in vivo in HLA‐A*0201‐transgenic HHD mice and in vitro in HLA‐A*0201+ healthy donors. p391‐ and p393‐specific human and murine CTL recognized human tumor cells overexpressing Hsp70 in a HLA‐A*0201‐restricted manner. Tetramer analysis of TILs showed that these Hsp70 epitopes are targets of an immune response in many HLA‐A*0201+ breast cancer patients. Hsp70 is a tumor antigen and the Hsp70‐derived peptides p391 and p393 could be used to raise a cytotoxic response against tumors of various origins. © 2003 Wiley‐Liss, Inc.
Exposure to cement dust, related occupational groups and laryngeal cancer risk: Results of a population based case‐control study
Authors: Dietz Andreas , Ramroth Heribert , Urban Tobias , Ahrens Wolfgang , Becher Heiko
Published: January 2004

A population‐based case‐control study was performed in the Rhein‐Neckar region, Germany, to evaluate occupational risk factors for the development of laryngeal cancer (“Rhein‐Neckar‐Larynx Study”). Between May 1998 and December 2000, 257 patients (236 males, 21 females), aged 37–80, with histologically confirmed laryngeal cancer, as well as 769 population control persons (702 males, 67 females), were included (1:3 frequency matched by age and sex). History of occupational exposures, as well as other risk factors (tobacco, alcohol), was obtained with face‐to‐face interviews using a detailed standardized questionnaire. The complete individual work history was assessed. A detailed assessment of work conditions was obtained by job‐specific questionnaires (JSQs) for selected jobs known to be associated with exposure to potential laryngeal carcinogens. Estimates for total exposure hours by substance were calculated based on JSQs. Published occupational hygiene data were used to infer semiquantitative scores of exposure intensity for specific job tasks. After adjustment for tobacco and alcohol intake, a significant elevated odds ratios (OR) could be demonstrated for persons that were exposed to cement during their work as building and construction workers. An OR of 2.42 was calculated for workers of the high exposed subgroup (95% confidence interval: 1.14‐5.15; p < 0.001). Smoking was the main confounding factor because the unadjusted cement OR of 3.20 dropped down to 2.42 after adjustment for tobacco intake. We conclude that there is good evidence for cement dust exposure acting as a tobacco, alcohol and asbestos independent risk factor for laryngeal carcinoma. Our study gives a base for further toxicologic investigations on this topic. © 2003 Wiley‐Liss, Inc.
The increase of female breast cancer incidence in Japan: Emergence of birth cohort effect
Authors: Minami Yuko , Tsubono Yoshitaka , Nishino Yoshikazu , Ohuchi Noriaki , Shibuya Daisuke , Hisamichi Shigeru
Published: January 2004

During recent decades, breast cancer incidence has been increasing in Japan. According to the latest reports from several cancer registries in Japan, the breast has become the leading cancer site in female cancer incidence. To analyze the trend of breast cancer incidence in detail, we summarized female breast cancer incidence in Miyagi Prefecture, Japan during 1959–1997, and evaluated the period and cohort effect on breast cancer incidence using the age‐period‐cohort model. Age‐specific and age‐standardized rates have increased over successive calendar periods. Around 1980, an accelerated increase in these incidence rates took place. A full model including age, period and cohort was best fitted to the trend of incidence. In the model, the effects of period and cohort were statistically significant. The nonlinear effect for cohort indicates an increasing trend, beginning with the cohort in 1888–1897, and the nonlinear effect for period showed a clear increase in risk with calendar period. Furthermore, the full model including a linear component showed a steadily upward trend in the cohort effect. Based on our own epidemiologic studies previously conducted in Miyagi Prefecture, and other published reports, the cohort effect is likely to be related to the change in prevalence of women with risk factors such as low parity and insufficient breastfeeding. We believe that the emergence of the cohort effect is an important finding, although the period effect may also persist. The significant cohort effect may give a caution for continuous increase of breast cancer incidence in Japan. © 2003 Wiley‐Liss, Inc.