bluuebunny/crossref_metadata_2025 huggingface dataset:
The development of tolerance to Clostridium perfringens type D o-toxin in MDCK and G-402 cells
Authors: Beal Dominic R , Titball Richard W , Lindsay Christopher D
Published: January 2004

The epithelial Madin Darby canine kidney (MDCK) cell line, Caucasian renal leiomyoblastoma (G-402) cells, human small airways epithelial (HSAE) cells, human bronchial epithelial (HBE) cells and human renal proximal tubule (HRPT) epithelial cells were examined for sensitivity to Clostridium perfringens biotype D o-toxin. MDCK and G-402 cells were confirmed as being the only established cell lines that are sensitive to the toxin. HSAE, HBE and HRPT epithelial cells were only found to be sensitive to the toxin at concentrations of -1 mg/ mL. Cultures of MDCK and G-402 cells, with increased resistance (tolerance) to the cytotoxic effects of o-toxin, were developed by exposing these cultures to progressively higher concentrations of toxin. The greatest relative increase in tolerance to o-toxin was developed in MDCK cells, in which the LC50 in control cultures was 2mg/mL as determined by the MTS/PMS assay system; after selection for tolerance, this was raised to 100 mg/mL. This represents a 50-fold increase in tolerance as measured by this index. Using G-402 cells, it was possible to increase the LC50 by twofold from 290 to 590 mg/mL.

Subsequent 2-D electrophoresis of membrane preparations from tolerant and control MDCK cells revealed that the expression of a discrete group of proteins found in control cells with a range of molecular weights from 32–36 kDa, all with acidic isoelectric points (IEPs), were either not expressed in o-toxin tolerant cells or had undergone a shift in IEP to a more alkaline pH in tolerant cells. This suggests that o-toxin lethality in MDCK cells may be mediated by membrane-located proteins. Their absence or alteration in toxin-resistant cells would, at least partly, explain the failure of most cell lines to demonstrate sensitivity to this toxin, despite being derived from tissues that are damaged by o-toxin. This approach may have utility in the study of other toxin / cell interactions and could be used in the development of novel medical countermeasures by identifying cellular targets which mediate toxin lethality.
Prothrombin time prolongation in paracetamol poisoning: a relevant marker of hepatic failure?
Authors: Payen C , Dachraoui A , Pulce C , Descotes J
Published: January 2004

The association between paracetamol overdose and prolonged prothrombin time due to hepatic failure is well recognized. However, little is known of the possibility that paracetamol overdose can prolong the prothrombin time without overt hepatic failure. The few data from the literature suggest this is either due to a reduction in the functional levels of the vitamin K-dependent clotting factors by elevated doses of paracetamol, or a consequence of the administration of the antidote N-acetylcystein. The three reported cases provide further evidence that paracetamol overdose can be associated with a prolongation in the prothrombin time without overt hepatic failure. Even though the prothrombin time provides useful prognosis information, decisions regarding the management of these patients should not solely be based on this endpoint to avoid misinterpretation of the accuracy and the severity of liver failure.
Protection by sildenafil and theophylline of lead acetate-induced oxidative stress in rat submandibular gland and saliva
Authors: Abdollahi Mohammad , Fooladian Fatemeh , Emami Bita , Zafari Khatereh , Bahreini-Moghadam Arezou
Published: January 2004

The role of oxidative stress in lead toxicity has been proposed in many organs, however, no study has been performed in the salivary glands, which are important parts of the gastrointestinal tract with a high implication in health of the whole body. Recently, it has been proposed that increasing the levels of cGMP and cAMP in the cells may protect from the neurotoxicity of lead. The objective of this study was to determine the ability of lead acetate to produce oxidative stress in rat submandibular as the main salivary gland of the body and to study the role of pretreatment by specific phosphodiesterase inhibitors in the prevention of oxidative stress. Lead acetate (100 mg/kg), alone or in combination with theophylline (25 mg/kg) and sildenafil (5 mg/kg), was administered intraperitoneally to rats. After 2 hours and under general anaesthesia, the submandibular gland ducts were cannulated intraorally using microcannula, and pure saliva was collected for 30 min using pilocarpine (8 mg/kg) as a secretagogue. The submandibular glands were then isolated free under surgery. Oxidative stress in the gland and pure saliva were evaluated measuring lipid peroxidation (thiobarbituric acid reactive substances assay), total thiol groups content and total antioxidant capacity (the ferric reducing ability assay). Results showed significant oxidative stress in the gland and secretions as indicated by increased lipid peroxidation, decreased total antioxidant capacity and thiol group levels. The use of cAMP and cGMP phosodiesterase inhibitors, theophylline and sildenafil, prevented leadinduced increased lipid peroxidation and also protected from decreased thiol groups content and total antioxidant power of the gland and secretions. The same trend of effects was observed in gland and saliva. It is concluded that lead toxicity is mediated through oxidative stress in salivary glands, while increasing intracellular cAMP and cGMP levels may prevent lead-induced oxidative stress.
Paraoxonase activity and genetic polymorphisms in greenhouse workers with long term pesticide exposure
Authors: Herńndez Antonio F , Mackness Bharti , Rodrigo Lourdes , López Olga , Pla Antonio , Gil Fernando , Durrington Paul N , Pena Gloria , Parrón Tesifón , Serrano José L , Mackness Michael I
Published: January 2004

Serum paraoxonase (PON1) is a high-density lipoprotein (HDL) associated protein, which plays a critical role in the pathogenesis of atherosclerosis, although it was primarily associated with the hydrolysis of organophosphorus compounds. PON1 was initially thought to be independent from physiological or pathological states, although recently some environmental factors have been reported to modulate its activity. In this study, we have investigated the promoter (PON1-108C/T and-909 C/G) and coding region (PON1 192Q/R and 55L/M) polymorphisms, as well as PON1 activity towards different substrates (paraoxon, phenylacetate and diazoxon) in 102 individuals with long term low dose exposure to pesticides in a plastic greenhouse setting (sprayers), who are probably the group of agricultural workers with the highest exposure to pesticides. PON1 activity towards paraoxon was nonsignificantly decreased (up to 53.5%) in the sprayers subgroup exposed to organophosphates (n-41) compared with nonsprayers acting as controls (n-39). None of the genotypes studied was associated significantly with the subgroup of individuals exposed to organophosphates, although differences between sprayers and nonsprayers were observed in the PON1-909 G/C polymorphism. Among the environmental factors that significantly predicted lower rates of PON1 activity towards paraoxon are, interestingly, the exposure to organophosphates and current smoking. By contrast, the utilization of protective clothing while spraying pesticides inside the greenhouses was positively associated with PON1 activity, very likely by preventing the pesticides from being absorbed. This study suggests that chronic exposure to pesticides might decrease PON1 activity and pinpoints the potential usefulness of monitoring PON1 activity in occupational settings where exposure to organophosphates occurs.
In vitro studies on the toxicity of isoniazid in different cell lines
Authors: Schwab Christina E , Tuschl Helga
Published: January 2004

The aim of the present study was to investigate in vitrothe mechanism of toxicity of isoniazid (-INH), the drug most widely used for treatment of tuberculosis. The human hepatoma line HepG2, the human lymphoblastoid line AHH-1 and the murine lymphoma cells YAC-1 were used as test systems. Active cell death (-apoptosis) and necrosis were detected by different flow cytometric methods: the binding of annexin V to the cell membrane and staining with propidium iodide (PI), the TUNEL assay for detection of DNA fragmentation and the occurrence of a sub G1 peak in cell cycle histograms. Mitochondrial membrane potential was analysed with the fluorescent probe JC-1. In addition to cytotoxicity, effects of INH on cell cycle were studied in HepG2 cells. The data of the present investigations indicate that INH induces cytotoxicity via apoptosis both in hepatoma and lymphoma cells. Twenty-four hours of application of INH in concentrations -26 mM led to a remarkable number of apoptotic cells positive for Annexin V. The induction of apoptosis was accompanied by a break down of the mitochondrial membrane potential and the occurrence of DNA strand breaks. At incubation times from 36 to 48 hours, a sub-G1 peak of late apoptotic cells was detected in cell cycle analysis. Furthermore, cell cycle studies showed a disruption of the cycle at low concentrations of INH which are only mildly cytotoxic. Thus the present study unequivocally demonstrated that INH induces cytotoxicity via apoptosis and can lead to a significant disturbance of the cell cycle in mammalian cells.
Language activation in monozygotic twins discordant for schizophrenia
Authors: Sommer Iris E. C. , Ramsey Nick F. , Mandl René C. W. , Van Oel Clarine J. , Kahn René S.
Published: January 2004

Background

In previous functional magnetic resonance imaging (fMRI) studies, participants with schizophrenia showed decreased language lateralisation, resulting from increased activation of the right hemisphere compared with controls.

Aim

To determine whether decreased lateralisation and increased right cerebral language activation constitute genetic predispositions for schizophrenia.

Method

Language activation was measured using fMRI in 12 right-handed monozygotic twin pairs discordant for schizophrenia and 12 healthy right-handed monozygotic twin pairs who were matched for gender, age and education.

Results

Language lateralisation was decreased in discordant twin pairs compared with the healthy twin pairs. The groups did not differ in activation of the language-related areas of the left hemisphere, but language-related activation in the right hemisphere was significantly higher in the discordant twin pairs than in the healthy pairs. Within the discordant twin pairs, language lateralisation was not significantly different between patients with schizophrenia and their co-twins.

Conclusions

Decreased language lateralisation may constitute a genetic predisposition for schizophrenia.
Contextual Assessment of the Maternity Experience: development of an instrument for cross-cultural research
Authors: Bernazzani Odette , Conroy Sue , Marks Maureen N. , Siddle Kathryn A. , Guedeney Nicole , Bifulco Antonia , Asten Paul , Figueiredo Barbara , Gorman Laura L. , Bellini Simona , Glatigny-Dallay Elisabeth , Hayes Sandra , Klier Claudia M. , Kammerer Martin H. , Henshaw Carol A. ,   TCS–PND Group
Published: January 2004

Background

There is evidence that stressors may trigger the onset of a depressive episode in vulnerable women. A new UK interview measure, the Contextual Assessment of the Maternity Experience (CAME), was designed to assess major risk factors for emotional disturbances, especially depression, during pregnancy and post-partum.

Aims

Within the context of a cros-scultural study, to establish the use fulness of the CAME, and to test expected associations of the measure with characteristics of the social context and with major or minor depression.

Method

The CAME was administered antenatally and postnatally in ten study sites, respectively to 296 and 249 women. Affective disorder throughout pregnancy and upto 6 month spostnatally was assessed by means of the Structured Clinical Interview for DSM–IV Axis I Disorders.

Results

Adversity, poor relationship with either a partner or a confidant, and negative feelings about the pregnancy all predicted onset of depression during the perinatal period.

Conclusions

The CAME was able to assess major domains relevant to the psychosocial context of the maternity experience in different cultures. Overall, the instrument showed acceptable psychometric properties in its first use in different cultural settings.
Surely you take complementary and alternative medicines?
Authors: Brugha Traolach , Rampes Hagen , Jenkins Rachel
Published: January 2004

A substantial proportion of our patients use or consider using complementary and alternative medicines (CAM) and other coping strategies. It is important that we acknowledge this, know something about the subject and are aware of current or potential developments in the field. These remedies might be harmless, beneficial or harmful and their side-effects might alter and confuse clinical presentations. We need to be vigilant of the potential for significant drug interactions between complementary and orthodox treatments. There is a substantial growth in complementary and alternative medical research in the USA, now beginning to follow in the UK. This will hopefully bring useful future progress.
The MRCPsych OSCE workshop: a new game to play?
Authors: Naeem Asim , Rutherford Joan , Kenn Chris
Published: January 2004

In line with the Royal College of Psychiatrists' commitment to improve its professional examinations based on the best current evidence, Spring 2003 has seen the introduction of the objective structured clinical examination (OSCE). This has replaced the individual patient assessment (IPA) in the MRCPsych Part I clinical examination. An OSCE consists of a series of time-limited clinical tasks that candidates have to perform in a consecutive series of ‘stations' or booths. They have the advantage of being able to test clinical competence using a number of different scenarios via a standardised format (Katona et al, 2000). Their use has become widespread over recent years, particularly in undergraduate psychiatry exams (Brewin & Cantwell, 1997), as they have good reliability and validity (Hodges et al, 1998). This was confirmed by the College's initial pilot OSCE, which had a κ score of around 0.8 for the examination as a whole (Oyebode, 2002).
Conceptual “Heat‐Driven” Approach to the Synthesis of DNA Oligonucleotides on Microarrays
Authors: GRAJKOWSKI A. , CIEŚLAK J. , CHMIELEWSKI M. K. , MARCHÁN V. , PHILLIPS L. R. , WILK A. , BEAUCAGE S. L.
Published: January 2004

Abstract: The discovery of deoxyribonucleoside cyclic N‐acylphosphoramidites, a novel class of phosphoramidite monomers for solid‐phase oligonucleotide synthesis, has led to the development of a number of phosphate protecting groups that can be cleaved from DNA oligonucleotides under thermolytic neutral conditions. These include the 2‐(N‐formyl‐N‐methyl)aminoethyl, 4‐oxopentyl, 3‐(N‐tert‐butyl)carboxamido‐1‐propyl, 3‐(2‐pyridyl)‐1‐propyl, 2‐[N‐methyl‐N‐(2‐pyridyl)]aminoethyl, and 4‐methythiobutyl groups. When used for 5′‐hydroxyl protection of nucleosides, the analogous 1‐phenyl‐2‐[N‐methyl‐N‐(2‐pyridyl)]aminoethyloxycarbonyl group exhibited excellent thermolytic properties, which may permit an iterative “heat‐driven” synthesis of DNA oligonucleotides on microarrays. In this regard, progress has been made toward the use of deoxyribonucleoside cyclic N‐acylphosphoramidites in solid‐phase oligonucleotide syntheses without nucleobase protection. Given that deoxyribonucleoside cyclic N‐acylphosphoramidites produce oligonucleotides with heat‐sensitive phosphate protecting groups, blocking the 5′‐hydroxyl of these monomers with, for example, the thermolabile 1‐phenyl‐2‐[N‐methyl‐N‐(2‐pyridyl)]aminoethyloxycarbonyl group may provide a convenient thermo‐controlled method for the synthesis of oligonucleotides on microarrays.
Modulation of Toll‐like Receptor 9 Responses through Synthetic Immunostimulatory Motifs of DNA
Authors: AGRAWAL SUDHIR , KANDIMALLA EKAMBAR R.
Published: January 2004

Abstract: Bacterial, plasmid, and synthetic DNA containing unmethylated CpG dinucleotides in specific sequence contexts activate the vertebrate innate immune system. A pattern recognition receptor (PRR), toll‐like receptor 9 (TLR9), recognizes CpG DNA and activates signaling cascade leading to the secretion of a number of cytokines and chemokines. Our extensive structure‐immunostimulatory activity relationship studies showed that a number of synthetic pyrimidine (Y) and purine (R) nucleotides are accepted by the receptor as substitutes for natural deoxycytidine and deoxyguanosine in a CpG dinucleotide. These studies permitted development of synthetic immunostimulatory motifs YpG, CpR, and YpR and established the nucleotide motif recognition pattern of the receptor. A number of site‐specific chemical modifications in the flanking sequences to the CpG dinucleotide permitted modulation of immunostimulatory affects in a predictable manner. Our studies also showed that TLR9 recognizes and reads the CpG DNA sequence from the 5′‐end. Design of oligonucleotides with two 5′‐ends, immunomers, resulted in potent immunomodulatory agents with distinct cytokine profiles. Immunomers containing synthetic immunostimulatory motifs produced different cytokine induction profiles compared with natural CpG motifs. Importantly, some of these synthetic motifs showed optimal activity in both mouse and human systems without requiring to change sequences, suggesting overriding the species‐dependent specificity of the receptor by the use of synthetic motifs. In this article, we review current understanding of structural recognition and functional modulation of TLR9 receptor by second‐generation immunomodulatory oligonucleotides and their potential application as wide spectrum therapeutic agents.
RNA‐Mediated Inhibition of HIV in a Gene Therapy Setting
Authors: MICHIENZI ALESSANDRO , CASTANOTTO DANIELA , LEE NANCY , LI SHIRLEY , ZAIA JOHN A. , ROSSI JOHN J.
Published: January 2004

Abstract: At present, treatment for HIV‐1 infection employs highly active anti‐retroviral therapy (HAART), which utilizes a combination of RT and protease inhibitors. Unfortunately, HIV can escape many therapies because of its high mutation rate and the complexity of its pathogenesis. HIV‐1 integrates into the cellular genome, which facilitates persistence and acts as a reservoir for reactivation and replication. As an alternative or adjuvant to chemotherapy we have been developing an RNA‐based gene therapy approach for the treatment of HIV‐1 infection. This article summarizes the various RNA based technologies that we have developed for potential application in a gene therapy setting.
CRE‐Enhancer DNA Decoy
Authors: CHO‐CHUNG YOON S.
Published: January 2004

Abstract: Enhancer DNA decoy oligonucleotides (ODNs) inhibit transcription by competing for transcription factors. A decoy ODN composed of the cAMP response element (CRE) inhibits CRE‐directed gene transcription and tumor growth without affecting normal cell growth. We used DNA microarrays to analyze the global gene expression in tumors exposed to the CRE‐decoy ODN. The CRE decoy upregulated the AP‐2β transcription factor gene in tumors but not in the livers of host animals. The upregulated expression of AP‐2β was clustered with other upregulated genes involved in development and cell differentiation. Concomitantly, another cluster of genes involved in cell proliferation and transformation was downregulated. The observed alterations indicate that CRE‐directed transcription favors tumor growth. Evidence presented here suggests that the CRE‐decoy ODN may provide a target‐based genetic tool for treating cancer, viral diseases, and other diseases in which CRE‐directed transcription is abnormally used.
Gene Targeting by Triple Helix‐Forming Oligonucleotides
Authors: MAJUMDAR ALOKES , PURI NITIN , McCOLLUM NICHOLAS , RICHARDS SALLY , CUENOUD BERNARD , MILLER PAUL , SEIDMAN MICHAEL M.
Published: January 2004

Abstract: Effective gene targeting reagents would have widespread utility for genomic manipulation including transgenic cell and animal construction and for gene therapy. They would also be useful in basic research as probes of chromatin structure, and as tools for studying the repair and mutagenesis of targeted DNA damage. We are developing triple helix‐forming oligonucleotides (TFOs) for gene targeting in living mammalian cells. Challenges to TFO bioactivity include the impediments to the biochemistry of triplex formation presented by the physiological environment and the charge repulsion between the duplex and the third strand. In addition, there are biological constraints to target access imposed by mammalian chromatin structure. Here we describe the oligonucleotide modification format that appears to support biological activity of TFOs. In addition we show that manipulation of the cell biology, specifically the cell cycle, has a dramatic influence on TFO bioactivity.
Imaging Oncogene Expression
Authors: TIAN XIAOBING , ARUVA MOHAN R. , RAO PONUGOTI S. , QIN WENYI , READ PAUL , SAUTER EDWARD R. , THAKUR MATHEW L. , WICKSTROM ERIC
Published: January 2004

Abstract: In 2003, approximately 39,800 women in the US will die from breast cancer. Mammography and physical examination miss up to 40% of early breast cancers. Moreover, if an abnormality is found, an invasive diagnostic procedure must still be performed to determine if the breast contains atypia or cancer, even though approximately 85% of abnormalities are benign. Scintigraphic imaging of gene expression in vivo by noninvasive means could direct physicians to appropriate targets for intervention at the onset of disease and thereby significantly impact patient management. Until now, no method has been available to image specific overexpressed oncogene mRNAs in vivo by scintigraphic imaging. We hypothesize that gamma‐emitting Tc‐99m‐PNA‐peptides can be taken up by human ER+ and ER− breast cancer xenografts, hybridize to complementary mRNA targets in those cells, and concentrate sufficiently in tumor tissue to allow noninvasive imaging of oncogene overexpression. To prepare the probes, peptide analogs of insulin‐like growth factor 1 (IGF1) were extended from a solid support by Fmoc coupling. Peptide nucleic acid (PNA) dodecamers antisense to CCND1 and MYC mRNAs were then extended from the N‐terminus of IGF1, followed by a chelator peptide, using Fmoc coupling for all residues. The cysteine thiols were cyclized on the solid support, either before or after PNA extension. This simplified synthetic approach allows preparation of a variety of multipeptide disulfide‐bridged PNA chimeras. A chelating peptide‐PNA chimera antisense to MYC mRNA was then labeled efficiently with Tc‐99m, yielding a single product. Tissue distribution studies of antisense and mismatch chimeras at 4 h and 24 h after administration displayed modest accumulation in the liver and kidneys, with appreciable levels in tumors. This result enables testing of Tc‐99m‐peptide‐PNA probes to image gene expression in tumors.
The Therapeutic Potential of Antigen‐Oligonucleotide Conjugates
Authors: DATTA SANDIP K. , TAKABAYASHI KENJI , RAZ EYAL
Published: January 2004

Abstract: Conjugation of protein antigen with immunostimulatory oligonucleotides creates a potent immunogen that elicits antigen‐specific antibody responses, a Th1‐biased cytokine profile, and CD8 cytotoxic T lymphocyte activity. The wide range of humoral and cellular immune responses induced by these conjugates suggests they can be used to create effective vaccines against infectious pathogens and tumors and to beneficially modulate the immune responses seen in allergic diseases. This review summarizes the available data on the use of antigen‐oligonucleotide conjugates and discusses their potential use for the treatment of infectious, oncologic, and allergic diseases in humans.
Applying Antisense Technology
Authors: HOLMLUND JON T.
Published: January 2004

Abstract: Progress in the development of antisense drugs over the last decade has led to the approval of the first such drug—Vitravene® for AIDS‐related CMV retinitis—and the development of a large number of antisense drugs in clinical trials. Antisense drugs are now being studied in Phase 3 trials for patients with cancer and inflammatory bowel disease. Other antisense drugs are in development for rheumatoid arthritis, other inflammatory conditions, and hepatitis C. Still other antisense drugs are entering clinical trials for treatment of metabolic conditions such as diabetes and hyperlipidemia. These latter applications provide the potential for target effects to be more directly measured in the clinic. Improved antisense chemistry, which will enhance the feasibility of subcutaneous and oral administration of antisense drugs and offer the potential of less frequent dosing, is expected to further expand the opportunities for antisense drug development.
Surveillance of Duodenal Adenomas in Familial Adenomatous Polyposis Reveals High Cumulative Risk of Advanced Disease
Authors: Saurin Jean-Christophe , Gutknecht Christelle , Napoleon Bertrand , Chavaillon Annick , Ecochard René , Scoazec Jean-Yves , Ponchon Thierry , Chayvialle Jean-Alain
Published: January 2004

Purpose

The development of high-grade dysplasia (HGD) on duodenal or jejunal adenomas and of late-stage (stage IV) duodenal polyposis are major clinical events for familial adenomatous polyposis (FAP) patients. Our aim was to determine their respective frequency, risk factors, and cumulative risk.

Patients and Methods

A prospective, optimized, endoscopic surveillance protocol was applied to 58 FAP patients in a university hospital. The number, size, and histology of duodenojejunal polyps were assessed, and the Spigelman’s score was calculated at each endoscopy. Cox regression and linear regression analysis were used to determine risk factors for HGD development and the cumulative risk of stage IV duodenal polyposis, respectively.

Results

During a median (± standard deviation) follow-up of 47.9 ± 15.6 months, 35 patients with at least two consecutive examinations had 107 duodenojejunal examinations. The Spigelman’s score increased in 21 patients (60.0%), and HGD developed in 12 patients (34.2%). High initial Spigelman’s score (> 7 points), but not age or APC mutation site, was a risk factor for HGD development. Estimated cumulative risk of developing stage IV duodenal polyposis was of 42.9% at age 60 (95% CI, 35.7% to 50.0%) and 50.0% at age 70 (95% CI, 42.9% to 57.1%).

Conclusion

This prospective series shows a higher duodenal polyposis progression rate and cumulative risk of late-stage (stage IV) duodenal polyposis in FAP patients compared with previous series. These results suggest that current modalities for surveillance and management of these patients need revision.
Effect of Duration of Treatment on Treatment Outcome for Patients With Clear-Cell Sarcoma of the Kidney: A Report From the National Wilms' Tumor Study Group
Authors: Seibel Nita L. , Li Sierra , Breslow Norman E. , Beckwith J. Bruce , Green Daniel M. , Haase Gerald M. , Ritchey Michael L. , Thomas Patrick R.M. , Grundy Paul E. , Finklestein Jerry Z. , Kim Tae , Shochat Stephen J. , Kelalis Panayotis P. , D'Angio Giulio J.
Published: January 2004

Purpose

To evaluate the effect of conventional and standard (ST) versus pulse-intensive (PI) chemotherapy and short-duration versus long-duration chemotherapy on relapse-free survival (RFS) and overall survival rates of patients with clear-cell sarcoma of the kidney (CCSK) entered onto the National Wilms' Tumor Study (NWTS)-4.

Patients and Methods

The 5-year and 8-year RFS rates were determined for patients with CCSK treated on the NWTS-4. After August 6, 1986, 40 previously untreated children younger than 16 years with CCSK were randomly assigned, after the completion of 6 months of chemotherapy, to discontinue (short) or continue 9 additional months (long) of treatment with chemotherapy regimens that included vincristine and either divided-dose (ST) courses (5 days) or single-dose (PI) treatment with dactinomycin and divided-dose (ST) courses (3 days) or single-dose (PI) treatment with doxorubicin.

Results

For patients with CCSK, the 5- and 8-year RFS rates were 65.2% and 60.6%, respectively, for patients randomly assigned to the short chemotherapy and 87.8% (both 5- and 8-year RFS) for patients randomly assigned to the long chemotherapy (P = .08). The overall survival rates for patients at 5 and 8 years were 95.5% and 85.9%, respectively, for the short chemotherapy and 87.5% (both 5- and 8-year overall survival) for the long chemotherapy (P = .99). In NWTS-4, the overall survival rates for patients with CCSK improved from NWTS-3 (83% v 66.9% at 8 years, respectively; P < .01).

Conclusion

CCSK patients exhibit an improved RFS from a longer course of therapy when using vincristine, doxorubicin, and dactinomycin, but their long-term survival is unchanged compared with patients receiving 6 months of therapy. The overall survival rates for patients with CCSK have improved from NWTS-3.
Combination of Cytology, Fluorescence In Situ Hybridization for Aneuploidy, and Reverse-Transcriptase Polymerase Chain Reaction for Human Mammaglobin/Mammaglobin B Expression Improves Diagnosis of Malignant Effusions
Authors: Fiegl Michael , Haun Margot , Massoner Anita , Krugmann Jens , Müller-Holzner Elisabeth , Hack Rene , Hilbe Wolfgang , Marth Christian , Duba Hans-Christoph , Gastl Günther , Grünewald Kurt
Published: January 2004

Purpose

The identification of malignant cells in effusions by conventional cytology is hampered by its limited sensitivity. The aim of this study was to improve tumor cell detection in effusions by molecular approaches.

Materials and Methods

A total of 157 effusions from patients with tumors and 72 effusions from patients without a history or evidence of malignancy were included in this study. All effusion specimens were evaluated in parallel by cytology, fluorescence in situ hybridization (FISH) for aneuploidy, and reverse-transcriptase polymerase chain reaction (RT-PCR) for expression of human mammaglobin (hMAM) and mammaglobin B (hMAM-B).

Results

In effusions from patients with tumors, the sensitivities of tumor cell detection by cytology, FISH, and hMAM and hMAM-B detection were 46.2%, 53.3%, 36.4%, and 57.7%, respectively. The corresponding specificities were 94.4%, 97.0%, 87.1%, and 88.6%. Notably, a high percentage of effusions containing malignant cells were in fact transudates, indicating the necessity for molecular diagnostic work-up of transudates collected from patients with tumors. Dependent on the tumor type, the use of appropriate marker combinations improved tumor cell detection in effusions significantly. By combining all four diagnostic tests, a positive test result indicating the presence of malignancy was achieved in 81.1%, with a fairly good specificity of 70.1%.

Conclusion

Molecular techniques are definitely useful to detect malignancy in cytologically negative effusions. Tumor cell detection in effusions can be significantly improved by FISH and PCR techniques applying appropriate molecular markers. This finding should help to improve tumor staging, prognostic assessment, and treatment monitoring.
Relapse-Free and Overall Survival in Patients With Pathologic Stage II Nonseminomatous Germ Cell Cancer Treated With Etoposide and Cisplatin Adjuvant Chemotherapy
Authors: Kondagunta G. Varuni , Sheinfeld Joel , Mazumdar Madhu , Mariani Tania V. , Bajorin Dean , Bacik Jennifer , Bosl George J. , Motzer Robert J.
Published: January 2004

Purpose

To assess the long-term relapse-free survival and overall survival of patients with stage II nonseminomatous germ cell tumor (NSGCT) who received two cycles of adjuvant etoposide and cisplatin (EP) after primary retroperitoneal lymph node dissection.

Patients and Methods

Eighty-seven patients with completely resected pathologic stage II NSGCT were treated with adjuvant EP chemotherapy. Adjuvant EP consisted of two cycles of etoposide (100 mg/m2) plus cisplatin (20 mg/m2) per day, administered days 1 to 5 at a 21-day interval.

Results

Ten patients (11%) had pN1 disease, 73 (84%) had pN2 disease, and four (5%) had pN3 disease. Eighty-six patients received two cycles of EP, and one patient received an additional two cycles of EP after a transient marker increase after his first cycle. Eighty-seven patients are alive, and 86 patients (99%) remain relapse-free at a median follow-up of 8 years (range, 0.9 to 13.5 years).

Conclusion

Two cycles of adjuvant EP is highly effective in preventing relapse in patients with pathologic stage II pN1 and pN2 NSGCT. An alternative treatment strategy is surveillance with full-course chemotherapy at relapse. Because there is a higher risk of relapse for patients with pN2 disease, these patients are offered adjuvant chemotherapy.
Functional characterization of active transport of progesterone to adrenal cells
Authors: Ogihara Takuo , Matsumoto Shigeki , Ohnishi Shuhei
Published: January 2004

The characterization of the transport mechanism of progesterone, which is one of the neutral steroids in the adrenal cells, has been studied by the examination of progesterone uptake into the monolayers of SW-13 cells (a human adrenal adenocarcinoma cell line). The uptake of [3H]progesterone at a tracer concentration (1 nM) exhibited temperature, pH and sodium dependency. According to kinetic analysis of the concentration dependence, the uptake of progesterone involves saturable and non-saturable processes. The uptake for the saturable process, which gave Kt values (half-saturation concentration) of 4.7 ± 8.7 μM, was inhibited by metabolic inhibitors and amino-acid modifiers but not by endocytosis inhibitors or substrates for known transporters. The uptake of progesterone was also inhibited by several neutral steroids but not by anionic steroids. The inhibition by both β-estradiol and estriol was competitive. The uptake of progesterone by the adrenal cells might be at least partially accounted for by a specific carrier-mediated transport mechanism generated by sodium ions and an electrochemical mechanism.
Quantitative characterization of direct P-glycoprotein inhibition by St John's wort constituents hypericin and hyperforin
Authors: Wang Er-jia , Barecki-Roach Mary , Johnson William W
Published: January 2004

The ATP-binding cassette transporter P-glycoprotein (P-gp) exerts a critical role in the systemic disposition of, and exposure to, lipophilic and amphipathic drugs, carcinogens, toxins and other xenobiotics. The ability of P-gp to transfer a wide variety of structurally unrelated compounds from the cell interior across the membrane bilayer remains intriguing. Since natural product chemicals in the widely consumed St John's wort appear to exert antidepressant effects by an unknown mechanism, the constituents are frequently studied for interactions with various biomacromolecules as well as cytotoxins or isolated cells. The drug interactions caused by this widely used herbal remedy are under-appreciated. Various clinical interactions have been observed upon the co-administration of St John's wort, and P-gp and CYP3A4 have been indicted as the cause. We characterized several St John's wort constituents for their interaction with P-gp and their specific effects on the P-gp export activity of several marker substrates. Two of these constituents, hyperforin and hypericin, inhibit the active efflux of the fluorescent markers daunorubicin (IC50 ˜ 30 μM) and calcein-AM. Herein, we show in-vitro results that can both explain the competing clinical observations of initial elevated exposure of P-gp substrate drugs (P-gp inhibition) followed by under-exposure (P-gp induction) when St John's wort is co-administered, and provide a further warning against unchecked co-administration of drugs with St John's wort.
Anti-diabetic activity of Zingiber officinale in streptozotocin-induced type I diabetic rats
Authors: Akhani Sanjay P , Vishwakarma Santosh L , Goyal Ramesh K
Published: January 2004

The fresh and dried rhizome of Zingiber officinale Roscoe (commonly known as ginger) is widely used in traditional medicine. We have studied the effect of the juice of Z. officinale (4 mL kg−1, p.o. daily) for 6 weeks on streptozotocin (STZ)-induced type I diabetic rats with particular reference to the involvement of serotonin (5-hydroxytryptamine; 5-HT) receptors in glycaemic control. In normoglycaemic rats, 5-HT (1 mg kg−1, i.p.) produced hyperglycaemia and hypoinsulinaemia, which was significantly prevented by the juice of Z. officinale. STZ-diabetes produced a significant increase in fasting glucose levels that was associated with a significant decrease in serum insulin levels. Treatment with Z. officinale produced a significant increase in insulin levels and a decrease in fasting glucose levels in diabetic rats. In an oral glucose tolerance test, treatment with Z. officinale was found to decrease significantly the area under the curve of glucose and to increase the area under the curve of insulin in STZ-diabetic rats. Treatment with Z. officinale also caused a decrease in serum cholesterol, serum triglyceride and blood pressure in diabetic rats. Our data suggest a potential antidiabetic activity of the juice of Z. officinale in type I diabetic rats, possibly involving 5-HT receptors.
Sonodynamic antitumour effect of chloroaluminum phthalocyanine tetrasulfonate on murine solid tumour
Authors: Yumita Nagahiko , Umemura Shin-ichiro
Published: January 2004

The sonodynamically induced antitumour effect of chloroaluminum phthalocyanine tetrasulfonate (AlPcTS) was evaluated on subcutaneously implanted colon 26 carcinoma. A time of 24h after the administration of AlPcTS was chosen for the ultrasonic exposure, based on the analysis of the AlPcTS concentrations in the tumour, plasma, skin and muscle. The pharmacokinetic analysis showed much faster clearance of AlPcTS than photofrin II from the body, which can be an advantage in view of their potential adverse effects. At an AlPcTS dose not less than 2.5 mg kg−1 and at a free-field ultrasonic intensity not less than 3 W cm−2, the synergistic effect between AlPcTS administration and ultrasonic exposure on the tumour growth inhibition was significant. The ultrasonic intensity showed a relatively sharp threshold for the synergistic antitumour effect, which is typical for an ultrasonic effect mediated by acoustic cavitation. These results suggest that AlPcTS is a potential sonosensitizer for sonodynamic treatment of solid tumours.
Loperamide modifies the tissue disposition kinetics of ivermectin in rats
Authors: Lifschitz Adrian L , Virkel Guillermo L , Sallovitz Juan M , Pis Alejandra , Imperiale Fernanda A , Lanusse Carlos E
Published: January 2004

Ivermectin (IVM) is a broad-spectrum antiparasitic drug extensively used in human and veterinary medicine that is largely excreted in bile and faeces. Loperamide (LPM) is an opioid derivative that reduces gastrointestinal secretions and motility. Both IVM and LPM have been reported to act as P-glycoprotein substrates (P-GP). The goal of the present work was to study the LPM-induced modifications to the pattern of tissue distribution for IVM. Thirty-six Wistar male rats were randomly allocated to two groups (n = 18) and treated subcutaneously with IVM alone or co-administered with LPM. Rats were killed at different times post-treatment and samples (blood and tissues) were collected and analyzed by HPLC. The presence of LPM induced a marked enhancement in the IVM plasma concentrations, resulting in a significantly higher area under concentration time curve (AUC) value (P < 0.01) than that obtained after the administration of IVM alone. Significantly higher IVM availabilities in the liver tissue and small intestine wall (P < 0.05) were obtained in the presence of LPM. There were no statistically significant differences in drug availability in the large intestinal wall after both treatments. However, LPM induced a marked decrease in the amount of IVM recovered in the large intestinal lumen content. The ratio between IVM concentrations in the large intestinal luminal content and plasma at day 1 post-treatment was 4.64-fold higher in the absence of LPM. The delayed intestinal transit time caused by LPM accounting for an extended plasma–intestine recycling time, and a potential competition between IVM and LPM for the P-GP-mediated bile–intestinal secretion processes, may account for the enhanced IVM systemic availability reported in the current study.
Ginsenoside-Rd attenuates oxidative damage related to aging in senescence-accelerated mice
Authors: Yokozawa Takako , Satoh Akiko , Cho Eun Ju
Published: January 2004

Among the various theories of the aging process, the free radical theory, which proposes that deleterious actions of free radicals are responsible for the functional deterioration associated with aging, has received widespread attention. The theory suggests that enhancement of the antioxidative defence system to attenuate free-radical-induced damage will counteract the aging process. We used senescence-accelerated mice (SAM) to investigate the relationship between aging and the antioxidative defence system and evaluated the effects of ginsenoside-Rd, the saponin from ginseng, by measuring antioxidative defence system parameters, including the glutathione (GSH)/glutathione disulfide (GSSG) redox status, antioxidative enzyme activity and level of lipid peroxidation. SAM at 11 months of age (old SAM) showed a significantly lower hepatic GSH/GSSG ratio, due to decreased GSH and increased GSSG levels, than SAM at 5 weeks of age (young SAM). However, the administration of ginsenoside-Rd at a dose of 1 or 5 mg kg−1 daily for 30 days to 10-month-old SAM significantly increased GSH, but decreased GSSG, resulting in elevation of the GSH/GSSG ratio. In addition, ginsenoside-Rd increased the activity of glutathione peroxidase (GSH-Px) and glutathione reductase that were both significantly lower in old SAM than in young SAM. This suggests that ginsenoside-Rd could play a crucial role in enhancing the defence system through regulation of the GSH/GSSG redox status. Moreover, decreases in the superoxide dismutase (SOD) and catalase activity in old SAM compared with young SAM were also revealed, indicating that the aging process resulted in suppression of the antioxidative defence system. However, ginsenoside-Rd did not affect SOD and catalase activity. As catalase is localized in peroxisome granules and GSH-Px is present in the cytoplasm and mitochondrial matrix, the site of ginsenoside-Rd action may be the cytoplasm and mitochondrial matrix. Furthermore, the serum and liver malondialdehyde levels, indicators of lipid peroxidation, were elevated with aging, while ginsenoside-Rd inhibited lipid peroxidation. This study indicates that the aging process leads to suppression of the antioxidative defence system and accumulation of lipid peroxidation products, while ginsenoside-Rd attenuates the oxidative damage, which may be responsible for the intervention of GSH/GSSG redox status.
Antihypertensive and anti-arrhythmic effects of an extract of Radix Stephaniae Tetrandrae in the rat
Authors: Yu X-C , Wu S , Chen C-F , Pang K T , Wong T M
Published: January 2004

In this study, we determined the effects of an extract of Radix Stephaniae Tetrandrae (RST) on arterial blood pressure and heart weight in deoxycorticosterone acetate-salt (DOCA-salt) hypertensive rats. We also determined the effects of the extract on arrhythmia and infarct induced by myocardial ischaemia and reperfusion in anaesthetized rats. We further compared the effects of the extract with those of tetrandrine, which makes up 7% of the extract and is known to act as a calcium-channel antagonist, and verapamil, a prototype calcium-channel antagonist. Treatment with RST extract returned the arterial blood pressure, cardiac compliance and coronary flow towards normal, and reduced right ventricular hypertrophy in the DOCA-salt hypertensive rat. In the anaesthetized rat, the RST extract reduced arrhythmia and infarct size induced by myocardial ischaemia and reperfusion; the effects were similar to those of tetrandrine and verapamil. The findings indicate that the RST extract acts like a calcium-channel antagonist. It may be used in the treatment of cardiovascular diseases, as are the calcium-channel antagonist and tetrandrine. More interestingly, the effects of the RST extract were of the same potency as tetrandrine. Since only 7% of the extract was tetrandrine, the observation indicates that tetrandrine was not the only component that was responsible for the actions of the extract.
Water extract isolated from Chelidonium majus enhances nitric oxide and tumour necrosis factor-α production via nuclear factor-κB activation in mouse peritoneal macrophages
Authors: Chung Hwan-Suck , An Hyo-Jin , Jeong Hyun-Ja , Won Jin-Hee , Hong Seung-Heon , Kim Hyung-Min
Published: January 2004

Chelidonium majus is used to treat several inflammatory diseases and tumours. We have examined the effect of C. majus on nitric oxide (NO) production using mouse peritoneal macrophages. When C. majus was used in combination with recombinant interferon-γ (rIFN-γ, 10U mL−1), there was a marked cooperative induction of NO production. Treatment of rIFN-γ plus C. majus (1 mg mL−1) in macrophages caused a significant increase in tumour necrosis factor-α (TNF-α) production. The increased production of NO and TNF-α from rIFN-γ plus C. majus-stimulated cells was almost completely inhibited by nuclear factor-κB (NF-κB) inhibitor, pyrrolidine dithiocarbamate (100 μM). These findings demonstrated that C. majus increased the production of NO and TNF-α by rIFN-γ-primed macrophages and suggested that NF-κB played a critical role in mediating the effects of C. majus.
Acute and topic anti-edematogenic fractions isolated from the seeds of Pterodon pubescens
Authors: Silva M C C , Gayer C R M , Lopes C S , Calixto N O , Reis P A , Passaes C P B , Paes M C , Dalmau S R , Sabino K C C , Todeschini A R , Coelho M G P
Published: January 2004

We previously demonstrated that alcoholic extracts from Pterodon pubescens Benth. (Sucupira branca, Leguminosae) seeds exhibit anti-arthritic activity. In the present work we show that the oleaginous extract obtained from P. pubescens seeds (OEP) exhibits acute or topic anti-edematogenic activity when tested in carrageenan-induced paw edema or in croton oil-induced ear edema assays, respectively. Four fractions were obtained from OEP by sequential liquid–liquid extraction. The anti-edematogenic properties were predominant in the hexanic fraction, which was further fractionated by HPLC, yielding three sub-fractions (PF1.1, PF1.2 and PF1.3). PF1.1 and PF1.3 showed potent acute and topic anti-edematogenic activity. The PF1.2 sub-fraction, although not active in the carrageenan assay, exhibited a potent anti-edematogenic activity in the croton oil-induced ear edema. This sub-fraction shows a maximum efficacy similar to indometacin in a lower dose. The PF1.1 sub-fraction presented a complex mixture containing furane diterpene derivatives of vouacapan. PF1.2 consists of a single substance, geranylgeraniol, as determined by GC/MS and NMR, while PF1.3 contains farnesol.