bluuebunny/crossref_metadata_2025 huggingface dataset:
Rhabdomyosarcoma in adults
Authors: Ferrari Andrea , Dileo Palma , Casanova Michela , Bertulli Rossella , Meazza Cristina , Gandola Lorenza , Navarria Pierina , Collini Paola , Gronchi Alessandro , Olmi Patrizia , Fossati‐Bellani Franca , Casali Paolo G.
Published: July 2003

BACKGROUND

The goal of the current study was to clarify treatment outcomes for adult patients with rhabdomyosarcoma (RMS). Published series have reported definitively worse results for adults with RMS compared with children with RMS. This finding casts doubt on whether RMS is the same disease in adults as it is in children.

METHODS

Of 190 patients with RMS who were age 18 years or older and whose cases were recorded over a 25‐year span in the pathology database of the Istituto Nazionale Tumori (Milan, Italy), 171 could be analyzed retrospectively for treatment outcome. The authors attempted to stratify patients according to the degree to which they had been treated appropriately, based on current treatment guidelines for childhood RMS.

RESULTS

The overall rate of response to chemotherapy was 85%. For the entire series, 5‐year event‐free survival and 5‐year overall survival (OS) were 28% and 40%, respectively. Among the 110 patients with embryonal, alveolar, or ‘not otherwise specified’ RMS, 5‐year OS was 46%; however, 5‐year OS was 61% for patients within this group (39% of the total) who had high scores for appropriate treatment.

CONCLUSIONS

The current series parallels other published series in that it confirms the finding of a relatively poor long‐term outcome for adult patients with RMS. However, for patients whose treatment adhered to the current guidelines for treatment of children, outcome was similar to what has been reported in pediatric series. In addition, the rate of response to chemotherapy for the entire series was similar to the rate typically observed among children. These findings suggest that adults and children with RMS should receive similar treatment. Treatment protocols adopted from pediatric programs but tailored to adults could increase adults' chances of receiving appropriate treatment; prospective studies are needed to test this idea. Cancer 2003;98:571–80. © 2003 American Cancer Society.

DOI 10.1002/cncr.11550
Levels of the soluble, 55‐kilodalton isoform of tumor necrosis factor receptor in bone marrow are correlated with the clinical outcome of children with acute lymphoblastic leukemia in first recurrence
Authors: Wu Shuling , Korte Alexander , Gessner Reinhard , Henze Guenter , Seeger Karlheinz
Published: July 2003

BACKGROUND

It has been shown that the soluble, 55‐kilodalton isoform of tumor necrosis factor receptor (sTNFRp55) enhances tumor survival by exhibiting competitive ligand binding. The objective of the current study was to determine the levels of sTNFRp55 and their impact on outcome in 106 children with acute lymphoblastic leukemia (ALL) in first recurrence.

METHODS

Between January 1997 and December 2001, bone marrow (BM) samples were collected from 106 children with a first recurrence of ALL at diagnosis. These patients were enrolled in the Berlin–Frankfurt–Münster (BFM) ALL recurrence trial, ALL‐REZ BFM 90‐96. Levels of sTNFRp55 in BM samples were determined with a commercially available enzyme‐linked immunosorbent assay kit. Event‐free survival (EFS) and overall survival were assessed from the date of study entry or the date of randomization, as appropriate.

RESULTS

The mean sTNFRp55 level (± standard deviation) was 3.40 ± 2.57 ng/mL. High levels of sTNFRp55 were associated with shorter duration of first complete remission and observation time as well as poor response to chemotherapy. Most importantly, the probability of EFS (pEFS) at 3 years was significantly worse for children with recurrent ALL who had sTNFRp55 levels greater than the median value (> 2.77 ng/mL) compared with patients who had levels that were less than the median value (pEFS: 0.44 ± 0.10 ng/mL vs. 0.12 ± 0.10 ng/mL; P = 0.006). It is noteworthy that the sTNFRp55 levels in 22 children with recurrent, TEL‐AML1‐positive ALL ([t(12;21)(p13;q22)]; 2.69 ± 1.05 ng/mL) were significantly lower compared with the levels in children who had TEL‐AML1‐negative ALL (3.34 ± 1.49 ng/mL; P < 0.05).

CONCLUSIONS

The results indicated that a high sTNFRp55 level represents a negative prognostic factor for children with recurrent ALL in terms of EFS and overall survival. Cancer 2003;98:625–31. © 2003 American Cancer Society.

DOI 10.1002/cncr.11553
Multiple primary tumors involving cancer of the brain and central nervous system as the first or subsequent cancer
Authors: Inskip Peter D.
Published: July 2003

BACKGROUND

This study was undertaken to determine whether cancer of the brain and central nervous system (CNS) occurs together with other types of cancer more often or less often than would be expected due to chance.

METHODS

The study was based on data collected by the Surveillance, Epidemiology, and End Results (SEER) Program for cancers diagnosed in the U.S. between 1973 and 1998. The standardized incidence ratio (SIR) for new malignancies was estimated as the number of patients diagnosed with a particular type of cancer (observed), divided by the number of diagnoses expected based on person‐years of follow‐up and incidence rates for SEER cancer registries.

RESULTS

Among patients who were diagnosed first with cancer of the brain or CNS, statistically significant excesses were observed for cancers of bone (SIR = 14.4), soft tissue (SIR = 4.6), brain and CNS (SIR = 5.9), salivary gland (SIR = 5.1), and thyroid gland (SIR = 2.7) in addition to acute myelocytic leukemia (SIR = 4.1) and melanoma of the skin (SIR = 1.7). Excess risk of new malignancies was markedly greater after first cancers of the brain or CNS diagnosed in childhood compared with similar diagnoses in adulthood. In reverse associations, significant excesses of cancer of the brain and CNS were observed only after diagnoses of acute lymphocytic leukemia (SIR = 7.4) and cancer of the testis (SIR = 1.8) or the thyroid gland (SIR = 1.7).

CONCLUSIONS

Cancer treatment appears to have been the major factor underlying most of the positive associations between brain cancer and primary cancers of other types, with a probable lesser contribution from shared genetic susceptibility. Results provide little or no evidence that brain cancer shares important etiologic factors with the common cancers of adulthood. Cancer 2003;98:562–70. Published 2003 by the American Cancer Society.

DOI 10.1002/cncr.11554
Oxidative stress is generated via the mitochondrial respiratory chain during plant cell apoptosis
Authors: Weir Iona E. , Pham Nhu‐An , Hedley David W.
Published: July 2003

Background

We present evidence that in plant cells DNA damage induced by the topoisomerase trapping drug camptothecin induces oxidative stress via the mitochondrial respiratory chain.

Methods

Flow cytometry was used to analyse mitochondrial respiratory chain activity by simultaneous measurement of mitochondrial generation of reactive oxygen intermediates (ROI) and mitochondrial membrane potential (ΔΨm), in live functioning sugarbeet protoplasts. The mitochondrial respiratory chain function was assessed by the addition of substrate and inhibitors to the digitonin‐permeabilised protoplasts. Mitochondrial activity was also measured in protoplasts at different stages of induced apoptosis by camptothecin.

Results

The addition of camptothecin to sugarbeet protoplasts resulted in increased mitochondrial generation of ROI, occurring concurrently with the hyperpolarisation of the mitochondrial inner membrane. Subsequently, there was a decrease in mitochondrial‐generated ROI, in association with a decrease in ΔΨm. These camptothecin‐treated digitonin‐permeabilised protoplasts could be re‐energised and ROI decreased by the addition of malic acid in the early stages of apoptosis induction, but not in the latter stages. There was a depolarisation of the mitochondrial inner membrane in the final stages of apoptosis.

Conclusions

These results are consistent with impaired adenine nucleotide exchange across the mitochondrial membrane, suggesting that similar mechanisms regulate plant apoptosis as those described for mammalian apoptosis. Cytometry Part A 54A:109–117, 2003. © 2003 Wiley‐Liss, Inc.
NF‐κB inhibitor sesquiterpene parthenolide induces concurrently atypical apoptosis and cell necrosis: Difficulties in identification of dead cells in such cultures
Authors: Pozarowski Piotr , Halicka Dorota H. , Darzynkiewicz Zbigniew
Published: July 2003

Background

Apoptosis and necrosis (“accidental cell death”) are distinct modes of cell death. The feature that often distinguishes apoptotic from necrotic cells is preservation of the plasma membrane integrity, reflected by ability of the former cells to exclude cationic dyes such as propidium iodide (PI) for a certain length of time. During necrosis, the plasma membrane is rapidly ruptured and necrotic cells stain intensely with PI. While studying cytostatic effects of the anti‐inflammatory sesquiterpene parthenolide (PRT), we have noticed that, concurrent with apoptosis, the cells were dying by necrosis in the same cultures. Furthermore, because apoptosis was atypical, reflected by rapid loss of plasma membrane integrity, it was difficult to distinguish apoptotic from necrotic cells based on this feature.

Methods

Three methods were used to distinguish apoptosis from necrosis: (a) HL‐60 cells treated with PRT were subjected to analysis of caspases activation using antibody that detects activated (cleaved) caspase‐3, (b) apoptotic cells were identified by binding of fluorochrome‐labeled inhibitor of caspases FAM‐VAD‐FMK combined with the PI exclusion assay, and (c) cellular DNA and RNA were differentially stained with acridine orange (AO).

Results

Apoptotic cells were characterized by (a) caspase‐3 activation detected immunocytochemically and (b) binding of FAM‐VAD‐FMK followed by or concurrent with (c) loss of ability to exclude PI, (d) deficit in DNA content, and (e) relatively little changed RNA content. Necrotic cells showed (a) no evidence of caspase‐3 activation, (b) no binding of FAM‐VAD‐FMK, (c) inability to exclude PI, (d) rapid loss of RNA, and (e) unchanged DNA content.

Conclusions

Identification of apoptotic cells versus necrotic cells was possible either based on the evidence of caspase‐3 activation, by labeling with FAM‐VAD‐FMK combined with PI or by differential staining of cellular DNA and RNA with AO. The data indicate that plasma membrane appears to be one of the targets of PRT, because its integrity is lost very early during cell death, which is reflected by atypical apoptosis and by primary necrosis (lysis of the membrane). Cytometry Part A 54A:118–124, 2003. © 2003 Wiley‐Liss, Inc.
Novel method for the isolation and characterisation of the putative prostatic stem cell
Authors: Bhatt Rupesh I. , Brown Michael D. , Hart Claire A. , Gilmore Paul , Ramani Vijay A.C. , George Nicholas J. , Clarke Noel W.
Published: July 2003

Background

Prostate stem cells, responsible for the development, maturation, and function of the prostate, have been implicated in the aetiology of both benign prostate hyperplasia (BPH) and prostate cancer (CaP). However, research has been hampered by the lack of a definitive stem cell marker. We have adapted the protocol for differential Hoechst 33342 uptake by hemopoietic stem cells to enable isolation of putative stem cells from the prostate.

Methods

Prostate epithelial cells isolated from prostate tissue obtained from patients with BPH after transurethral resection of the prostate were stained with Hoechst 33342. The Hoechst 33342 Red/Blue flow cytometry profile was then determined. Hoechst 33342 and Pyronin Y staining was used to determined the cell cycle status.

Results

A verapamil‐sensitive side population (SP) can be isolated from primary prostate tissue accounting for 1.38% ± 0.07% of prostate epithelial cells. Cell cycle analysis of this SP population revealed that the majority of SP cells are in either G0 (12.38 ± 0.31%) or G1 (63.19 ± 2.13%).

Conclusions

The Hoechst 33342 dye efflux protocol can be adapted for the isolation of a SP from primary prostate tissue. Cytometry Part A 54A:89–99, 2003. © 2003 Wiley‐Liss, Inc.
Sensitive and reliable JC‐1 and TOTO‐3 double staining to assess mitochondrial transmembrane potential and plasma membrane integrity: Interest for cell death investigations
Authors: Zuliani Thomas , Duval Raphaël , Jayat Chantal , Schnébert Sylviane , André Patrice , Dumas Marc , Ratinaud Marie‐Hélène
Published: July 2003

Background

Apoptosis is currently studied by flow cytometry with mitochondrial membrane potential (Δψmt) and membrane integrity fluorochromes. Rhodamine 123 and DiOC6(3) remain controversial to identify cells displaying a low Δψmt. JC‐1 constitutes a good Δψmt indicator, due to a fluorescence shift from green to orange emission, according to the increase in Δψmt. Nevertheless, it is not feasible to analyze it simultaneously with propidium iodide. Among available fluorescent probes, TOTO‐3 seems to be a good candidate for double staining with JC‐1.

Methods

Cell death of HaCaT cells was induced by H2O2 and FasL. Samples were stained with DiOC6(3)/IP or JC‐1/TOTO‐3 then analyzed by flow cytometry. Results were supported by confocal microscopy analyses of mitochondrial membrane potential. Moreover, cell morphology was determined on the sorted subpopulations defined on the basis of staining (JC‐1 versus TOTO‐3).

Results

We found that JC‐1 is a more efficient mitochondrial probe than DiOC6(3). After stress induction, the fluorescence level of JC‐1 and TOTO‐3 clearly defined three fluorescent subpopulations, respectively: (1) JC‐1high and TOTO‐3low, (2) JC‐1low and TOTO‐3medium, and (3) JC‐1low and TOTO‐3high. Their morphologic aspects after cell sorting indicated that they corresponded to three functional states (intact, apoptotic, and necrotic cells), and data were supported by caspase activity measurements.

Conclusions

We propose a reliable and efficient staining, with JC‐1 and TOTO‐3 to discriminate three functional cellular states: intact, apoptotic, and necrotic/late apoptotic cells by flow cytometry. Cytometry Part A 54A:100–108, 2003. © 2003 Wiley‐Liss, Inc.
An aperture‐shifting light‐microscopic method for rapidly quantifying positions of cells in 3D matrices
Authors: Burton Kevin
Published: July 2003

Background

Rapid measurements of large numbers of cells in 3D are often required for measurements of cell migration. A method is presented for quantifying the position of cells in three‐dimensional gels using brightfield microscopy.

Methods

Images were recorded using transmitted light with a closed condenser aperture diaphragm positioned off axis to produce oblique illumination. Two or more images, each at the same focal plane, were obtained with the aperture shifted equally to either side of the optical axis. All in‐focus objects were at the same position in the two images, but out‐of‐focus objects were displaced parallel to the aperture movement.

Results

The method was tested using gels containing 12‐μm‐diameter glass beads or cells that had migrated several hundred micrometers into the gel. The position in the image plane varied linearly with axial position, being reversed for objects above and below the focal plane. Beads and cells could be visualized up to a depth of >1 mm in gels.

Conclusions

The method facilitates measurements of positions of cells in 3D matrices by eliminating the need to perform optical sectioning and can be used with any brightfield microscope.
Cytometry Part A 54A:125–131, 2003. © 2003 Wiley‐Liss, Inc.
Immunoreactivity of Stat5 phosphorylated on tyrosine as a cell‐based measure of Bcr/Abl kinase activity
Authors: Jacobberger James W. , Sramkoski R. Michael , Frisa Phyllis S. , Ye Peggy Peng , Gottlieb Megan A. , Hedley David W. , Shankey T. Vincent , Smith Bradley L. , Paniagua Mary , Goolsby Charles L.
Published: July 2003

Background

Stat51 (Signal Transducer and Activator of Transcription 5) is normally phosphorylated and activated by Janus kinases. In cells transformed with BCR/ABL, Stat5 is constitutively activated by promiscuous phosphorylation. Cytometry of intracellular antigens can be used to evaluate cell treatments affecting gene expression, because it precisely provides the fraction of affected cells and the quantitative change in expression. Here, we asked whether we could measure a phosphorylated epitope on Stat5 by cytometry, and whether that measurement would respond to Bcr/Abl inhibition.

Methods

Chronic myelogenous leukemia (CML) cell lines or control Bcr/Abl‐negative cells were treated or not with imatinib mesylate, fixed and permeabilized with formaldehyde followed by methanol; reacted with rabbit polyclonal and mouse monoclonal antibodies against an epitope including tyrosine 694 of Stat5a (pSTAT5); reacted with antibodies that mark mitotic cells; counterstained with secondary fluorescent antibodies and 4′,6‐diamidino‐2‐phenylindole (DAPI); and then subjected to flow cytometry. Western blotting was performed with pSTAT5 and Stat5 antibodies.

Results

Optimal fixation and staining parameters were established for pSTAT5 antibodies with K562 cells. These cells displayed high levels of immunoreactivity with pSTAT5 probes that could be inhibited uniformly with imatinib mesylate in a dose‐response and time‐dependent manner. The IC50 for downregulation of pSTAT5 immunoreactivity for K562 cells by cytometry was ∼70 nM. The inhibition half‐time was approximately 1 min. At micromolar doses this reactivity remained minimal for up to 7 days. Cultured cells also displayed a population of negative cells that increased under conditions related to cessation of cell growth (media nutrient depletion). This study also showed quantitatively that a rabbit polyclonal antibody that cross‐reacted with an additional epitope could be used successfully as a measure of Bcr/Abl activity.

Conclusion

We have developed a sensitive cytometric assay for Bcr/Abl kinase activity in human hematopoietic cell lines. Cytometry Part A 54A:75–88, 2003. © 2003 Wiley‐Liss, Inc.
Nerve collagens from diabetic and nondiabetic Sprague–Dawley and biobreeding rats: an atomic force microscopy study
Authors: Wang H. , Layton B. E. , Sastry A. M.
Published: July 2003

Background

Alterations in rat's nerve collagens due to diabetes may be related to the permanence of damage due to diabetic neuropathy. We (1) provide a methodology for determining the diameters of collagen fibers accounting for atomic force microscope (AFM) imaging artifacts, (2) present data on structural differences in sciatic nerve endoneurial, epineurial and tail tendon collagens of control and diabetic Sprague–Dawley and BioBreeding rats, and (3) compare results with literature values.

Methods

We measured collagen diameters and band spacing on endoneurial and epineurial sciatic nerve tissue, and tail tendon, in control and diabetic rats (STZ‐induced 12‐week diabetic SD and 16‐week spontaneously diabetic BB rats). We also developed a model to interpret the raw AFM data.

Results

All types of fibrillar collagen diameters studied became larger for diabetic versus control animals. Values for diabetic and control collagen fiber diameters in SD rats were 78 nm and 72 nm for SN epineurium, and 49 nm and 43 nm for SN endoneurium. For diabetic and control BB rats, these values were 83 nm and 77 nm (SN epineurium) and 49 nm and 43 nm (SN endoneurium). Values of 161 nm and 125 nm were found for diabetic and control tail tendon of BB rats. No significant changes were observed in any of the five comparisons made in D‐band spacings that ranged from 63 to 69 nm.

Conclusions

The best means we have found to reduce raw AFM data is to measure several diameters with a single scan, using valley‐to‐valley measurements. Structural, fibrillar collagens of the nerve and tendon become larger in rats exposed to prolonged diabetes. Copyright © 2003 John Wiley & Sons, Ltd.
Cardioprotection with pioglitazone is abolished by nitric oxide synthase inhibitor in ischemic rabbit hearts–comparison of the effects of pioglitazone and metformin
Authors: Kawabata Hitoshi , Ishikawa Kinji
Published: July 2003

Background

The effects of two drugs representing different classes of antidiabetic pharmacology (pioglitazone, a thiazolidinedione; and metformin, a biguanide) on the myocardial metabolism in the ischemia are poorly understood.

Methods

To test the hypothesis that cardioprotection of pioglitazone and metformin is associated with nitric oxide (NO), we studied the high energy phosphate metabolism by 31P‐nuclear magnetic resonance (NMR) in isolated rabbit hearts. Forty‐five minutes of continuous normothermic global ischemia was carried out. Pioglitazone or metformin was administered at the beginning, 60 min prior to the global ischemia, with or without the nitric oxide synthase inhibitor, L‐NAME, administered 5 min or 60 min prior to the ischemia. In the first experiment, whether NO was produced or not by administration of pioglitazone, for the prevention of myocardial ischemic injury, was investigated. Hearts of male Japanese white rabbits were divided into 4 experimental groups: the control (C) group, the P group consisting of pioglitazone treatment, the P + L5 group consisting of pioglitazone treatment with L‐NAME (5 min before ischemia), and the P + L60 group consisting of pioglitazone treatment with L‐NAME (60 min before ischemia). In the next experiment, a comparison between the effects of pioglitazone and metformin in preventing ischemic injury were studied. The hearts were divided into 4 experimental groups: the control (C) group, the P group consisting of pioglitazone treatment, the P + L5 group consisting of pioglitazone treatment with L‐NAME (5 min before ischemia), the M group consisting of metformin treatment, and the M + L5 group consisting of metformin treatment with L‐NAME (5 min before ischemia).

Results

In the first experiment, the decrease in adenosine triphosphate (ATP) during ischemia was significantly inhibited in the P group in comparison with the C group (P < 0.01). However, the decrease in ATP was not inhibited in the P + L5 group during ischemia. In contrast, in the P + L60 group, the decrease in ATP was not inhibited during a part of ischemia. In the next experiment, a comparison between the effects of pioglitazone and metformin in preventing ischemic injury was studied. As a result of administration of either pioglitazone or metformin, there was no difference between groups with and without L‐NAME.

Conclusion

These results suggest that pioglitazone has a significant beneficial effect on improving the myocardial energy metabolism during ischemia. This cardioprotection may be dependent on nitric oxide (NO) synthase during ischemia more than preischemia. Furthermore, the present findings suggest that both pioglitazone and metformin have equal cardioprotective effects mediated by NO on myocardial ischemic injury in rabbits. Copyright © 2003 John Wiley & Sons, Ltd.
Assessment of insulin sensitivity in vivo in control and diabetic mice with a radioactive tracer of glucose transport: [125I]‐6‐deoxy‐6‐iodo‐D‐glucose
Authors: Perret P. , Ghezzi C. , Mathieu J. P. , Morin C. , Fagret D.
Published: July 2003

Background

Impairment of insulin‐stimulated glucose transport is a characteristic of type 2 diabetes. A radioactive glucose analogue has been synthesized: [125I]‐6‐deoxy‐6‐iodo‐D‐glucose. Its biological behaviour in vitro is similar to that of 3‐O‐methyl‐D‐glucose, the reference tracer of glucose transport. The aim of the present study was to determine the ability of [125I]‐6‐deoxy‐6‐iodo‐D‐glucose to evaluate variations in glucose transport in vivo.

Methods

Biodistributions of [125I]‐6‐deoxy‐6‐iodo‐D‐glucose were performed with or without exogenous insulin (iv injection of 1.5 IU/kg) in db/+ non‐diabetic control mice and in db/db type 2 diabetic mice, exhibiting a severe insulin resistance characterized by a lack of increase in glucose uptake in response to insulin.

Results

In db/+ mice, insulin increased [125I]‐6‐deoxy‐6‐iodo‐D‐glucose transport by 30% in most insulin‐sensitive tissues (heart, diaphragm and skeletal muscle, p < 0.05) and had no effect in other organs. In db/db mice, [125I]‐6‐deoxy‐6‐iodo‐D‐glucose transport in these organs was not modified by insulin.

Conclusion

[125I]‐6‐deoxy‐6‐iodo‐D‐glucose is able to trace in vivo an increase in glucose transport with insulin in non‐diabetic mice and a defect of glucose transport in type 2 diabetic mice. It is the first time that an iodinated analogue of glucose has shown such promising results after in vivo injection. The use of this tracer to assess glucose transport in vivo in humans via nuclear imaging warrants further investigation. Copyright © 2003 John Wiley & Sons, Ltd.
Early cell‐specific changes in nitric oxide synthases, reactive nitrogen species formation, and ubiquitinylation during diabetes‐related bladder remodeling
Authors: Poladia Deepali Pitre , Bauer John Anthony
Published: July 2003

Background

Urinary bladder dysfunction in diabetes is a well‐recognized phenomenon but the mechanisms involved and initiating events are not clear. The physiological production of nitric oxide (NO) plays an important role in bladder tone and local immune defense, and recent studies have shown that NO‐derived reactive nitrogen species occur in many settings of chronic disease, including other diabetic complications. Here, we investigated the early time‐dependent and cell‐specific changes in the nitric oxide synthase isoforms (NOS1, 2, and 3), peroxynitrite, and ubiquitinylation in the well‐documented streptozotocin‐induced rat model of diabetes.

Methods

Immunohistochemical methods and automated digital imaging were used for the measurement of morphometric and histochemical analysis of the bladder tissue regions. Region‐specific 3‐nitrotyrosine (a biomarker of NO dysregulation and reactive nitrogen species formation) and ubiquitinylated protein prevalence (marker of proteasomal activity) were also investigated.

Results

Immunohistochemistry revealed early, time‐dependent, and cell‐specific alterations in the three isoforms of NOS. We also observed region‐specific increases in protein nitration, demonstrating first‐time evidence of reactive nitrogen species formation in this setting. The changes in nitration did not pattern changes in NOS2 induction or tissue ubiquitinylation, and these alterations preceded any detectable changes in bladder structure (3 days vs 2 weeks) in this same animal preparation.

Conclusions

These data demonstrate that selective and regionally distinct changes in nitric oxide production and impaired nitric oxide control are early events during diabetic cystopathy and that mechanisms leading to increased oxidative stress and proteasomal activation may be key participants leading to organ dysfunction in this setting. Copyright © 2003 John Wiley & Sons, Ltd.
Diabetic atrial fibrillation patients: mortality and risk for stroke or embolism during a 10‐year follow‐up
Authors: Klem Igor , Wehinger Cornelius , Schneider Barbara , Hartl Elisabeth , Finsterer Josef , Stöllberger Claudia
Published: July 2003

Background

To compare in atrial fibrillation patients with and without diabetes, (1) baseline characteristics, (2) additional risk factors for stroke or peripheric or visceral embolism (hypertension, previous stroke, age >75 years), (3) mortality, (4) stroke or embolism, and (5) oral anticoagulation in the year 2000.

Methods

Included were 409 outpatients with nonrheumatic atrial fibrillation (62 ± 12 years, 36% female). All underwent transthoracic and transesophageal echocardiography. Patients with thrombi received oral anticoagulation; patients without thrombi received aspirin until the follow‐up in 1995; afterwards, oral anticoagulation according to risk factors for stroke or embolism was recommended. Patients were contacted during the year 2000.

Results

Type 2 diabetes was diagnosed in 73 patients (18%). Sixteen (22%) diabetic and 169 (50%) nondiabetic patients had no other risk factors for stroke or embolism (p < 0.0001). Diabetic patients were older, had more frequent heart failure, hypertension, myocardial infarction, left ventricular dysfunction, valvular abnormalities, left atrial or appendage thrombi, larger left atria, and left atrial appendages than nondiabetic patients. Mean follow‐up was 115 months. Diabetic patients had a higher mortality than nondiabetic patients (7%/year versus 4%/year, p < 0.0001). The rate of stroke or embolism of diabetic (3%/year) and nondiabetic patients (2%/year) was similar. The rate of oral anticoagulation was higher in diabetic than in nondiabetic patients (p = 0.0066).

Conclusions

Diabetic patients with atrial fibrillation frequently have additional risk factors for stroke or embolism, and thus should be treated with oral anticoagulation. Whether in the rare cases of atrial fibrillation, in whom diabetes is the only clinical risk factor, oral anticoagulation is indicated cannot be answered by the present study. Copyright © 2003 John Wiley & Sons, Ltd.
Insulin secretion and action show differences in impaired fasting glucose and in impaired glucose tolerance in Asian Indians
Authors: Snehalatha C. , Ramachandran A. , Sivasankari S. , Satyavani K. , Vijay V.
Published: July 2003

Background

The aim of the study was to compare the abnormalities in insulin secretion and insulin sensitivity in impaired fasting glucose (IFG) and in impaired glucose tolerance (IGT).

Methods

Glucose tolerance was determined by oral glucose tolerance test in 289 non‐diabetic subjects selected for this study (M : F, 160 : 129, mean age 42.2 ± 11.0 (SD) years, mean body mass index (BMI) 24.9 ± 4.2 kg/m2). Diabetic subjects were excluded from the study. The association of insulin resistance and β‐cell function (calculated by HOMA model) with IFG and IGT was studied using linear and multiple logistic regression analyses. Subjects with normoglycaemia (NGT) were used as controls.

Results

Insulin resistance was significantly more in IFG and in IGT than in NGT. β‐cell dysfunction was significantly higher in IFG than in IGT. IFG was associated with both insulin resistance (OR = 9.11, p < 0.001) and β‐cell dysfunction (inverse correlation, OR = 0.103, p < 0.001), while only insulin resistance was significantly associated with IGT (OR = 1.24, p = 0.005).

Conclusions

In urban South Asian Indians, IFG differed from IGT in having combined insulin resistance and β‐cell dysfunction, while IGT was associated only with insulin resistance. Copyright © 2003 John Wiley & Sons, Ltd.
Intermediate metabolism in normal pregnancy and in gestational diabetes
Authors: Di Cianni G. , Miccoli R. , Volpe L. , Lencioni C. , Del Prato S.
Published: July 2003

Complex though integrated hormonal and metabolic changes characterize pregnancy. In the face of progressive decline in insulin action, glucose homeostasis is maintained through a compensatory increase in insulin secretion. This switches energy production from carbohydrates to lipids, making glucose readily available to the fetus. This precise and entangled hormonal and metabolic condition can, however, be disrupted and diabetic hyperglycemia can develop (gestational diabetes). The increase in plasma glucose level is believed to confer significant risk of complications to both the mother and the fetus and the newborn. Moreover, exposition of fetal tissues to the diabetic maternal environment can translate into an increased risk for development of diabetes and/or the metabolic syndrome in the adult life. In women with previous gestational diabetes, the risk of developing type 2 diabetes is greatly enhanced, to the point that GDM represents an early stage in the natural history of type 2 diabetes. In these women, accurate follow‐up and prevention strategies are needed to reduce the subsequent development of overt diabetes. This paper will review current knowledge on the modifications occurring in normal pregnancy, while outlining the mechanisms. In this paper, we will review the changes of intermediary metabolism occurring during pregnancy. In particular, we will outline the mechanisms responsible for gestational diabetes; the link between these alterations and associated maternal and neonatal morbidity will be examined. Copyright © 2003 John Wiley & Sons, Ltd.
The role of Lay Review Committees in diabetes research
Authors: Stenger David P.
Published: July 2003

Type 1 diabetes is unique among disease entities in having a large voluntary health nonprofit organization (the Juvenile Diabetes Research Foundation) that employs the process of review by laypersons (following a review by scientists) in selecting the recipients of its funding awards to individual investigators/trainees: grants, career‐development awards, fellowships, and ‘innovative grants.’ Therefore, that organization can be a suitable model on which an examination of lay review can be based. This paper summarizes (1) the history of how lay review originated and (2) this foundation's experience with it, (3) assesses the impact of the procedure on the discipline of diabetes science, and (4) examines the role it might play in the future, given the current state of that discipline. Copyright © 2003 John Wiley & Sons, Ltd.
Statin therapy for the treatment of diabetic dyslipidemia
Authors: Haffner Steven M.
Published: July 2003

The increasing prevalence of type 2 diabetes is a major problem for healthcare providers globally, since it is associated with serious microvascular and macrovascular complications. Although microvascular complications can be largely reduced with strict glycemic control, prevention of macrovascular disease involves a multifaceted approach that addresses all major risk factors, including dyslipidemia, hypertension, and insulin insensitivity. In particular, the treatment of diabetic dyslipidemia is a major challenge for diabetologists and cardiologists, as it is characterized by an array of lipid abnormalities. The management of diabetic dyslipidemia should initially include lifestyle approaches such as improved nutrition and weight reduction; however, the majority of patients require the addition of pharmacotherapy. Whilst insulin and/or oral hypoglycemic drugs are generally prescribed for the treatment of hyperglycemia, the addition of lipid‐lowering drugs may be necessary for the control of diabetic dyslipidemia. The American Diabetes Association guidelines recommend lowering of low‐density lipoprotein cholesterol (LDL‐C) as a first priority. Hydroxy‐methylglutaryl coenzyme A reductase inhibitors (statins) are recommended for first‐line therapy in diabetic patients, since these agents are effective at reducing LDL‐C levels. Whilst statins provide effective control of dyslipidemia in the majority of patients, more efficacious treatment regimens would provide greater benefit to more patients. Combination therapies may provide one solution to obtaining maximal lipid profile modifications, although the introduction of new, more efficacious agents for use as monotherapy may provide a more acceptable option, as drug combinations are often associated with poor tolerability and patient compliance. Copyright © 2003 John Wiley & Sons, Ltd.
Synthesis and Structure of New Macrocycles Including Spiro‐1,3‐Dioxane Units
Authors: Grosu Ion , Bogdan Elena , Plé Gérard , Toupet Loïc , Ramondenc Yvan , Condamine Eric , Peulon‐Agasse Valérie , Balog Mirela
Published: July 2003

The efficient synthesis of new macrocycles — “monomers” and “dimers” — containing spiro‐1,3‐dioxane units is reported. The structure of the compounds was determined from high‐field NMR spectra, FAB and MALDI+ mass spectrometry investigations. The solid‐state molecular structures of three compounds was determined. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Total Synthesis of Anhydromarasmone
Authors: Jauch Johann , Wallner Carsten , Herdtweck Eberhardt
Published: July 2003

A synthesis of anhydromarasmone is presented using, as key step, an iridium‐catalyzed double‐bond isomerization from a homoallyl ether to an allyl ether. Other transition metal complexes employed for double‐bond transposition resulted in the vinyl ether as the final product. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Photoinduced Charge Separation in Cyclohexylidene‐Based Donor−(σ‐Bridge)−Acceptor Compounds − Building Blocks for Materials
Authors: Oosterbaan Wibren D. , van Gerven Paul C. M. , van Walree Cornelis A. , Koeberg Mattijs , Piet Jacob J. , Havenith Remco W. A. , Zwikker Jan W. , Jenneskens Leonardus W. , Gleiter Rolf
Published: July 2003

We describe the syntheses and properties of five semi‐rigid donor−(σ‐bridge)−acceptor (D−σ−A) compounds. These compounds, which are candidates for incorporation in materials, have aniline‐type donors and (mixed) cyano‐ and alkoxycarbonyl‐1,1‐difunctionalized vinyl acceptors that are separated by a four‐σ‐bond bridge. Their electronic properties were studied by means of photoelectron spectroscopy in combination with ab initio MO calculations, cyclic voltammetry, UV‐Vis absorption spectroscopy, (time‐resolved) fluorescence spectroscopy, and time‐resolved microwave conductivity (TRMC). We found that these D−σ−A compounds exhibit a weak intramolecular charge‐transfer absorption band that borrows its intensity from local transitions. Upon photoexcitation, a charge‐separated state is generated in virtually quantitative yield. Conformational contraction or “folding” of this state takes place for the two D−σ−A compounds with a dicyanovinyl acceptor in apolar solvents. The absence of folding in compounds with an alkoxycarbonyl‐functionalized acceptor is attributed to a reduced lifetime of the charge‐separated state. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Chemical Contributions to Understanding Heparin Activity: Synthesis of Related Sulfated Oligosaccharides
Authors: Poletti Laura , Lay Luigi
Published: July 2003

Heparin and heparan sulfate are complex polysaccharides that modulate several biological events through the recognition of a number of distinct proteins. Chemical synthesis is a powerful tool to identify the unique binding domains responsible for the specific interactions and to generate potent analogues of the natural sequences. The present review reports the synthetic efforts of the last decade that are focused on the preparation of tailored fragments of these two polymers and analogues thereof. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Solid Phase Synthesis of Vancomycin Mimics
Authors: Arnusch Christopher J. , Pieters Roland J.
Published: July 2003

A solid phase synthesis of a model system of the DE ring system of vancomycin is described. The synthesis involved biocatalytic resolutions of unnatural amino acids, is compatible with conventional solid phase peptide synthesis and contains as the key step: an on‐bead SNAr cyclization. Binding of a cyclic peptide to the carboxylate of N‐Ac‐D‐Ala was demonstrated. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Studies of the Biogenesis of Verrucosins, Toxic Diterpenoid Glycerides of the Mediterranean Mollusc Doris verrucosa
Authors: Fontana Angelo , Tramice Annabella , Cutignano Adele , d’Ippolito Giuliana , Renzulli Licia , Cimino Guido
Published: July 2003

The biogenesis of verrucosins, diterpenoid glycerides of the marine mollusc Doris verrucosa, has been investigated by feeding experiments with labelled precursors. Incorporation of radioactive D‐[U‐14C]‐glucose into the diterpenes proved the de novo origin of verrucosins in the mollusc. Biogenesis of glycerol and terpenoid substructures has been investigated by feeding experiment with 13C‐labelled precursors. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Basicity and Solvatochromism of Concave Pyridines with Extended π‐Systems in Protic and Nonprotic Solvents
Authors: Storm Ole , Lüning Ulrich
Published: July 2003

Arylalkyne substituents have been connected to the 4‐positions of concave pyridines to give substituted concave pyridines 3, which possess absorption maxima above 300 nm in their UV spectra. Their protonation and their interaction with polar solvents have been studied. Firstly, with strong acids, protonation occurs, and relative basicities (logKass) have been determined. Substitution at the remote aryl ring influences the basicity of the pyridine, and Hammett plots show a linear relation between logKass and the electron density of the remote aryl ring. Secondly, concave pyridines 3 are capable of “sensing” hydrogen bonds between their pyridine nitrogen atom and an alcohol. Shifts of the absorption maxima to longer wavelengths are observed when certain quantities of alcohol are added. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Synthesis and UV/Vis Spectra of J‐Aggregating 5,5′,6,6′‐Tetrachlorobenzimidacarbocyanine Dyes for Artificial Light‐Harvesting Systems and for Asymmetrical Generation of Supramolecular Helices
Authors: Pawlik Andreas , Ouart André , Kirstein Stefan , Abraham Hans‐Werner , Daehne Siegfried
Published: July 2003

A new class of dyes, in which the self‐assembling property of surfactants is combined with the capability for light energy propagation over long distances in dye J‐aggregates, is described. This has been achieved by the syntheses of achiral 5,5′,6,6′‐tetrachlorobenzimidacarbocyanine dyes 1, possessing systematically varied hydrophobic and hydrophilic substituents at their nitrogen atoms. These substituents are introduced into the 5,6‐dichlorobenzimidazole precursor 3 by substitution, either firstly by nucleophilic reaction with ψ‐bromoalkylnitriles and secondly by quaternization with alkyl bromides (route II) or firstly by nucleophilic reaction with alkyl bromides and secondly by quaternization with ψ‐bromoalkylnitriles or ψ‐bromoalkyl esters (route III) and subsequent saponification. The UV/Vis spectra of 20 differently substituted dyes containing the same chromophore 1 have been investigated. The spectra of the dye monomers in dimethyl sulfoxide are nearly identical, with no signs of optically activity, whereas in aqueous alkaline solutions quite different spectra are obtained for the dyes, indicating the formation of different aggregates depending on the nitrogen substituents. One of these types of J‐aggregate is optically inactive and displays a single red‐shifted (with respect to the dye monomers) absorption band, resembling the behaviour of J‐aggregates of common cyanine dyes. In the cases of strongly amphiphilic 5,5′,6,6′‐benzimidacarbocyanines 1 with 1,1′‐dialkyl substituents longer than hexyl and 3,3′‐bis(2‐carboxyethyl), 3,3′‐bis(3‐carboxypropyl) or 3,3′‐bis(3‐sulfopropyl) substituents, a new type of J‐aggregate is formed, and is distinguished by a doubly or even triply split J‐absorption band that displays optical activity. A third type of aggregate showing different spectral behaviour occurs when the dyes contain very short 3,3′‐bis(carboxymethyl) substituents or strongly hydrophobic fluorinated octyl groups, or when all four nitrogen atoms are identically substituted by hydrophilic 3‐carboxypropyl groups. The various types of dye aggregates have been characterized through UV/Vis spectroscopic parameters such as the positions and widths of the absorption and fluorescence bands, the Stokes′ shifts, the coupling constants, and the strength of the J‐band splitting. The results provide new prospects for the development of new artificial light‐harvesting systems as well as for the understanding of the evolution of asymmetry in the biosphere. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Peptide‐Cavitands Based on Resorc[4]arenes − Synthesis and Structure
Authors: Berghaus Carsten , Feigel Martin
Published: July 2003

Amino acids and peptides were coupled to the four aminomethyl groups of the bridged resorc[4]arene 2, resulting in peptide cavitands 3 of the general structure 2 [−Aa1‐[Aa2‐[Aa3]]−N(H)R]4. These compounds contain either four amino acids (3a−3d: Aa1 = Gly, Val, Leu or Phe, R = H; 3e, 3f: Aa1 = Gly or Val, R = Z), four dipeptides (3g−3l: Aa1 = Gly, Aa2 = Gly, Val, Leu, Phe, Lys(Boc) or Pro, R = Z), four tripeptides (3m, 3n: Aa1 = Gly, Aa2 = Val, Aa3 = Val or Met, R = Z) or four β‐Ala dipeptides (3o, 3p: Aa1 = β‐Ala, Aa2 = Val or Leu, R = Z). The dipeptide and tripeptide cavitands 3g−3n, which have a glycine bonded to the resorcarene core, form stable inclusion complexes with acetonitrile in chloroform solution. The free energy of complexation exceeds −5.9 kcal mol−1. The exchange between complexed and free acetonitrile molecules is slow on the NMR spectroscopic time scale (298 K). NMR spectroscopic data suggest that the peptides in the acetonitrile complexes of 3g−3n form a cyclic array of hydrogen bonds at the upper rim involving the glycine CO and NH groups. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Recent Developments in the Ring‐Chain Tautomerism of 1,3‐Heterocycles
Authors: Lázár László , Fülöp Ferenc
Published: July 2003

Some important results from the past seven years on the ring‐chain tautomerism of 1,3‐heterocycles are reviewed, including substituent effects on the tautomeric equilibria and synthetic applications of this phenomenon. The structures and reactivities of numerous five‐ and six‐membered, saturated, N‐unsubstituted 1,3‐X,N‐heterocycles (X = O, S, NR) can be characterized by the ring‐chain tautomeric equilibria of the 1,3‐X,N‐heterocycles and the corresponding Schiff bases; this is often exploited advantageously in different areas of organic synthesis, and also in physical, medicinal and peptide chemistry. The selectivity of certain transformations can be explained on the basis of the ring‐chain tautomeric equilibration of the intermediates, followed by a shift in the equilibrium. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Synthesis of Dihalo‐Substituted Analogues of Tröger’s Base from ortho‐ and meta‐Substituted Anilines
Authors: Hansson Anna , Jensen Jacob , Wendt Ola F. , Wärnmark Kenneth
Published: July 2003

For the first time, ortho‐ and meta‐halo‐substituted anilines were successfully condensed with formaldehyde to dihalo‐substituted analogues of Tröger’s base. By using paraformaldehyde and TFA, yields of 2−85% of these potential supramolecular building blocks were obtained. Even the inconceivable achievement of condensing anilines unsubstituted in para‐position to analogues of Tröger’s base was successful. Adding our present results to our previous, makes it now possible to synthesize analogues of Tröger’s base halo‐substituted in almost any desired position in each of its two aromatic rings. In addition the first X‐ray structure of a dihalo‐substituted analogue of Tröger’s base, 3,9‐dibromo‐4,10‐dimethyl‐6H,12H‐5,11‐methanodibenzo[b,f][1,5]diazocine (17), is presented. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Synthesis of 2,6‐Disubstituted Piperidine‐Bearing α‐Amino Acid Side‐Chains
Authors: Boeglin Damien , Heitz Annie , Martinez Jean , Fehrentz Jean‐Alain
Published: July 2003

A straightforward synthesis of 2,6‐disubstituted piperidines bearing α‐amino acid side‐chains was developed. Synthesis was based on a Horner−Wadsworth−Emmons condensation of a β‐ketophosphonate derived from an α‐amino acid residue with a β‐homologated aldehyde of anN‐protected amino acid residue. The generated α‐β unsaturated ketones were then reduced, deprotected, and cyclized in a hydrogenation/hydrogenolysis one‐pot procedure to yield piperidine moieties. Introduction of a new conformational restriction in the obtained molecules allowed total assignment of the stereocenters by NMR experiments. This assignment allowed us to propose a mechanistic pathway during the cyclization process, explaining the loss of exocyclic carbon‐center configuration. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)