Showing papers by "Charité published in 1999"

Crucial role of the interleukin 1 receptor family member T1/ST2 in T helper cell type 2-mediated lung mucosal immune responses.

Abstract: T1/ST2 is an orphan receptor of unknown function that is expressed on the surface of murine T helper cell type 2 (Th2), but not Th1 effector cells. In vitro blockade of T1/ST2 signaling with an immunoglobulin (Ig) fusion protein suppresses both differentiation to and activation of Th2, but not Th1 effector populations. In a nascent Th2-dominated response, anti-T1/ST2 monoclonal antibody (mAb) inhibited eosinophil infiltration, interleukin 5 secretion, and IgE production. To determine if these effects were mediated by a direct effect on Th2 cells, we next used a murine adoptive transfer model of Th1- and Th2-mediated lung mucosal immune responses. Administration of either T1/ST2 mAb or T1/ST2-Ig abrogated Th2 cytokine production in vivo and the induction of an eosinophilic inflammatory response, but failed to modify Th1-mediated inflammation. Taken together, our data demonstrate an important role of T1/ST2 in Th2-mediated inflammatory responses and suggest that T1/ST2 may prove to be a novel target for the selective suppression of Th2 immune responses.

A genomewide analysis provides evidence for novel linkages in inflammatory bowel disease in a large European cohort.

Abstract: Inflammatory bowel disease (IBD) is characterized by a chronic relapsing intestinal inflammation, typically starting in early adulthood. IBD is subdivided into two subtypes, on the basis of clinical and histologic features: Crohn disease and ulcerative colitis (UC). Previous genomewide searches identified regions harboring susceptibility loci on chromosomes 1, 3, 4, 7, 12, and 16. To expand our understanding of the genetic risk profile, we performed a 9-cM genomewide search for susceptibility loci in 268 families containing 353 affected sibling pairs. Previous linkages on chromosomes 12 and 16 were replicated, and the chromosome 4 linkage was extended in this sample. New suggestive evidence for autosomal linkages was observed on chromosomes 1, 6, 10, and 22, with LOD scores of 2.08, 2.07, 2.30, and 1.52, respectively. A maximum LOD score of 1.76 was observed on the X chromosome, for UC, which is consistent with the clinical association of IBD with Ullrich-Turner syndrome. The linkage finding on chromosome 6p is of interest, given the possible contribution of human leukocyte antigen and tumor necrosis-factor genes in IBD. This genomewide linkage scan, done with a large family cohort, has confirmed three previous IBD linkages and has provided evidence for five additional regions that may harbor IBD predisposition genes.

Atypical multidrug resistance: breast cancer resistance protein messenger RNA expression in mitoxantrone-selected cell lines.

Abstract: Background Human cancer cell lines grown in the presence of the cytotoxic agent mitoxantrone frequently develop resistance associated with a reduction in intracellular drug accumulation without increased expression of the known drug resistance transporters P-glycoprotein and multidrug resistance protein (also known as multidrug resistance-associated protein). Breast cancer resistance protein (BCRP) is a recently described adenosine triphosphate-binding cassette transporter associated with resistance to mitoxantrone and anthracyclines. This study was undertaken to test the prevalence of BCRP overexpression in cell lines selected for growth in the presence of mitoxantrone. Methods Total cellular RNA or poly A+ RNA and genomic DNA were isolated from parental and drug-selected cell lines. Expression of BCRP messenger RNA (mRNA) and amplification of the BCRP gene were analyzed by northern and Southern blot hybridization, respectively. Results A variety of drug-resistant human cancer cell lines derived by selection with mitoxantrone markedly overexpressed BCRP mRNA; these cell lines included sublines of human breast carcinoma (MCF-7), colon carcinoma (S1 and HT29), gastric carcinoma (EPG85-257), fibrosarcoma (EPF86-079), and myeloma (8226) origins. Analysis of genomic DNA from BCRP-overexpressing MCF-7/MX cells demonstrated that the BCRP gene was also amplified in these cells. Conclusions Overexpression of BCRP mRNA is frequently observed in multidrug-resistant cell lines selected with mitoxantrone, suggesting that BCRP is likely to be a major cellular defense mechanism elicited in response to exposure to this drug. It is likely that BCRP is the putative "mitoxantrone transporter" hypothesized to be present in these cell lines.

Evidence for Distinct Substrate Specificities of Importin α Family Members in Nuclear Protein Import

Abstract: Heavy trafficking between the nucleus and the cytoplasm takes place in eucaryote cells. Various substrates, such as different forms of RNA and many nuclear proteins, must be exported into the cytoplasm. Other proteins, such as transcription factors and ribonucleoprotein particles, must be imported into the nucleus. Nuclear transport occurs through the nuclear pore complexes (NPCs), which are about 125 MDa in size (34). Whereas smaller molecules up to 20 to 60 kDa may passively diffuse through the NPCs into the nucleus, the import of larger substrates is generally receptor mediated (12). This import process depends on the presence of specific signal sequences within the substrate. Different types of import signals exist. One such signal consists of the so-called nuclear localization signals (NLSs), which are mainly characterized by clusters of basic amino acids, predominately lysines. Depending on the numbers of their charged clusters, the NLSs may be classified into monopartite and bipartite NLSs (8, 9). Several soluble factors of the classical nuclear protein import pathway have been identified so far, including the small GTPase Ran/TC4 (26, 30), importin α (16, 20, 45), importin β (1, 4, 14, 19, 39), and NTF2 (31, 36), which is involved in the import and export of Ran (41). Importin α functions as an adapter molecule by binding importin β via its amino-terminally located importin β binding (IBB) domain (13, 46) and by binding NLS-bearing proteins via its two NLS binding sites in the central area (5, 18). Importin β is the transport receptor that carries the importin α-NLS complex from the cytoplasm into the nuclear side of the NPC (17). Once inside the nucleus, importin β binds to RanGTP, which is generated within the nucleus by the chromatin-bound RanGDP/GTP exchange factor RCC1. This binding of importin β to RanGTP leads to the dissociation of the import complex (15). Whereas importin β is thought to return to the cytoplasm rapidly without other soluble factors, the export of importin α is mediated by its nuclear export factor CAS, which binds to importin α preferentially in the presence of RanGTP (24). In the cytoplasm, the importins are set free for another round of import by the concerted action of RanGAP1 and RanBP1. Only one gene coding for importin β has been identified in the organisms analyzed thus far. In contrast to what was found for importin β, several isoforms of importin α in humans have been described. These include importin α1/Rch1 (7, 45), importin α5/hSRP1 (6), importin α3/Qip1 (23, 42), importin α4/hSRP1γ (23, 32), importin α6 (23), and, here, the newly reported importin α7. Importin α7 is the human homologue of the recently identified mouse importin α-S2 (44). Based on the sequence similarity, the importin α proteins can be grouped into three subfamilies. Members of different subfamilies have about 50% sequence identity. Within one subfamily, the identity is at least 80%. Whereas several of these isoforms are also found in invertebrates, the yeast Saccharomyces cerevisiae has only one gene for importin α, SRP1. Why so many importin α isoforms exist in higher eucaryotes has not yet been definitely answered. Although there is some tissue specificity in the expression of these proteins (23, 32, 38, 44), most isoforms are expressed within the same tissues. Initial data indicate that there may be distinct substrate specificities of different importin α family members (11, 28, 33, 43). However, other data show that different importin α proteins can interact with the same substrate (37, 38). We compared all of the ubiquitously expressed human importin α proteins, Xenopus importin α2, and yeast SRP1p in their efficiencies to promote the nuclear import of different substrates. When testing only one substrate per assay, we found that most substrates (NLS-bovine serum albumin [BSA], nucleoplasmin, P/CAF, and hnRNP K) were imported by all importin α proteins with only marginal differences. The exception was the nuclear import of RCC1, which was efficient only with importin α3, not with other isoforms. If two substrates are offered at the same time, the various importin α proteins show striking differences in their substrate-specific import efficiencies.

Mutant NDUFV1 subunit of mitochondrial complex I causes leukodystrophy and myoclonic epilepsy.

Abstract: Mutant NDUFV1 subunit of mitochondrial complex I causes leukodystrophy and myoclonic epilepsy

Circulating levels of matrix metalloproteinases MMP-3 and MMP-1, tissue inhibitor of metalloproteinases 1 (TIMP-1), and MMP-1/TIMP-1 complex in rheumatic disease. Correlation with clinical activity of rheumatoid arthritis versus other surrogate markers.

Abstract: Objective. To investigate whether plasma levels of matrix metalloproteinases 3 (MMP-3, stromelysin), MMP-1 (collagenase), tissue inhibitor of metalloproteinases I (TIMP-1), and MMP-1/TIMP-1 complex (MT complex) are specifically elevated in erosive joint diseases compared to nonerosive rheumatic diseases, and to assess how these markers reflect the clinical activity of rheumatoid arthritis (RA) compared to circulating cytokines and markers of connective tissue turnover as well as established variables [C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and rheumatoid factor titer]. Methods. Plasma levels of MMP-3, MMP- 1, TIMP-1, and MT complex were determined by ELISA. One hundred fifteen patients with RA, 20 with osteoarthritis (OA), 28 with psoriasis arthritis (PsA), 24 with ankylosing spondylitis (AS), 3 groups with systemic autoimmune diseases, and 30 healthy controls were analyzed. In patients with RA routine laboratory variables, circulating inflammatory cytokines [interleukin I (IL-1), tumor necrosis factor-α (TNF-α), and IL-6], collagen degradation products, and markers of bone formation were determined in parallel and were correlated to 4 variables of clinical activity. Results. MMP-3 levels were markedly elevated in RA compared to controls and OA, but also in all other groups, including 26 patients with systemic lupus erythematosus (SLE). MMP-I levels were significantly elevated in RA, but also in OA, PsA, SLE, and mixed connective tissue disease. In contrast, MT complex was elevated in RA only. TIMP- I was not different from controls. CRP levels, MMP-3, and ESR correlated best with clinical activity of RA. In contrast, there was no correlation of IL- 1 and TNF-α and only a weak correlation of IL-6 with clinical measures. Among variables of connective tissue turnover, only pyridinoline and deoxypyridinoline crosslinks were weakly correlated with disease activity. Conclusion. Elevated MMP-3 and MMP- I levels are not specific for RA or for erosive joint diseases in general. In contrast, elevated MT complex levels were observed in patients with RA. However, the correlation of MT- 1 with clinical data was weaker than that of MMP-3. Elevated MMP-3 levels reflected disease activity of RA better than cytokine levels or markers of connective tissue turnover. However, MMP-3 levels do not exceed the association of CRP with clinical activity.

Phylogenetic analysis of an anaerobic, trichlorobenzene-transforming microbial consortium

Abstract: A culture-independent phylogenetic survey for an anaerobic trichlorobenzene-transforming microbial community was carried out. Small-subunit rRNA genes were PCR amplified from community DNA by using primers specific for Bacteria or Euryarchaeota and were subsequently cloned. Application of a new hybridization-based screening approach revealed 51 bacterial clone families, one of which was closely related to dechlorinating Dehalobacter species. Several clone sequences clustered to rDNA sequences obtained from a molecular study of an anaerobic aquifer contaminated with hydrocarbons and chlorinated solvents (Dojka et al., Appl. Env. Microbiol. 64:3869-3877, 1998).

Telomere shortening triggers a p53-dependent cell cycle arrest via accumulation of G-rich single stranded DNA fragments

Abstract: It has been repeatedly suspected that telomere shortening might be one possible trigger of the p53-dependent cell cycle arrest, although the mechanism of this arrest remained unclear. Telomeres in human cells under mild oxidative stress accumulate single-strand damage faster than interstitial repetitive sequences. In MRC-5 fibroblasts and U87 glioblastoma cells, which both express wild-type p53, oxidative stress-mediated production of single-strand damage in telomeres is concomitant to the accumulation of p53 and p21 and to cell cycle arrest. This response can be modeled by treatment of cells with short single stranded telomeric G-rich DNA fragments. The arrest is transient in U87 cells. Recovery from it is accompanied by up-regulation of telomerase activity and elongation of telomeres. Overexpression of mutated p53 is sufficient to reverse the phenotype of inhibition as well as the delayed activation of telomerase. These data suggest that the production of G-rich single stranded fragments during the course of telomere shortening is sufficient to trigger a p53 dependent cell cycle arrest.

Cellular sources of enhanced brain-derived neurotrophic factor production in a mouse model of allergic inflammation.

Abstract: The aim of this study was to investigate production and cellular sources of brain-derived neurotrophic factor (BDNF) production in allergic asthma. For this purpose a mouse model of chronic and severe ovalbumin (OVA)-induced airway inflammation was developed. Allergen-exposed mice developed elevated immunoglobulin E titers; airway inflammation with influx of lymphocytes, monocytes, and eosinophils; and airway hyperresponsiveness. In addition to an influx of inflammatory cells, interleukin (IL)-4 and IL-5 production were enhanced, macrophages showed morphologic signs of activation, and airway epithelium was thickened and displayed a goblet-cell hyperplasia with a marked mucus production. BDNF was detected using in situ hybridization and enzyme-linked immunosorbent assay. Constitutive expression of BDNF messenger RNA (mRNA) was observed in the respiratory epithelium of sensitized and nonsensitized mouse lungs. In addition, BDNF mRNA was detected in airway inflammatory infiltrations and bronchoalveolar lavage fluid (BALF) cells of OVA-sensitized and aerosol-challenged mice. Highest BDNF protein levels were detected in BALF after long-term allergen aerosol exposure. Analysis of BDNF production by isolated lymphocyte subsets revealed T but not B cells as a cellular source of BDNF. In addition, activated alveolar macrophages were identified as BDNF-positive cells. These data indicate that in allergic airway inflammation BDNF production is upregulated and immune cells serve as a source of BDNF.

Th1/Th2 balance in infection.

Abstract: Cytokines produced by T helper (Th) cells are of critical importance for the outcome of many infectious diseases. Producing the “right” set of Cyokines in response to an infectious agent can be a matter of life or death. The Th1/Th2 dichotomy, although an oversimplification has proven useful in the analysis of immune responses to infections. In some infectious diseases, most notably leishmaniasis or infections with gastrointestinal helminths, one Th subset is indispensable for clearing the infection, whereas the opposite Th subset is detrimental. More frequently, both Th1 and Th2 responses are required at different time points to effectively eradicate an infectious agent. The granuloma responses to eitherMycobacterium tuberculosis orSchistosoma mansoni provide illustrative examples and are discussed in this review. There is accumulating evidence for frequent coexpression of Th1 and Th2 Cyokines during the in vivo immune response to infections. The mechanisms by which infectious agents modulate Th1/Th2 phenotype development are summarized here. Finally, we review here the current evidence for cytokine imbalances induced by infections as pathogenic or protective factors in autoimmunity and allergy.

Death deflected: IL-15 inhibits TNF-alpha-mediated apoptosis in fibroblasts by TRAF2 recruitment to the IL-15Ralpha chain.

Abstract: Interleukin-15 (IL-15) is a potent inhibitor of several apoptosis pathways. One prominent path toward apoptosis is the ligand-induced association of TNF receptor 1 (TNFR1) with death domain adaptor proteins. Studying if and how IL-15 blocks TNFR1-mediated apoptosis in a murine fibroblast cell line (L929), we show here that IL-15 blocks TNFR1-induced apoptosis via IL-15Ralpha chain signaling. The intracellular tail of IL-15Ralpha shows sequence homologies to the TRAF2 binding motifs of CD30 and CD40. Most important, binding of IL-15 to IL-15Ralpha successfully competes with the TNFR1 complex for TRAF2 binding, which may impede assembly of key adaptor proteins to the TNFR1 complex, and induces IkappaBalpha phosphorylation. Thus, IL-15Ralpha chain stimulation is a powerful deflector of cell death very early in the apoptosis signaling cascade, while TNF-alpha and IL-15 surface as major opponents in apoptosis control.

Contaminations Occurring in Fungal PCR Assays

Abstract: Successful in vitro amplification of fungal DNA in clinical specimens has been reported recently. In a collaboration among five European centers, the frequency and risk of contamination due to airborne spore inoculation or carryover contamination in fungal PCR were analyzed. The identities of all contaminants were specified by cycle sequencing and GenBank analysis. Twelve of 150 PCR assays that together included over 2,800 samples were found to be contaminated (3.3% of the negative controls were contaminated during the DNA extraction, and 4.7% of the PCR mixtures were contaminated during the amplification process). Contaminants were specified asAspergillus fumigatus, Saccharomyces cerevisiae, and Acremonium spp. Further analysis showed that commercially available products like zymolyase powder or 10× PCR buffer may contain fungal DNA. In conclusion, the risk of contamination is not higher in fungal PCR assays than in other diagnostic PCR-based assays if general precautions are taken.

Maternal viral load and vertical transmission of HIV-1 an important factor but not the only one

Abstract: OBJECTIVES To investigate the association between maternal RNA load, risk of vertical transmission of HIV-1, and other variables. METHODS Plasma or serum samples from mothers of 373 children, enrolled in the prospective European Collaborative Study, were collected around time of delivery, and HIV-RNA quantified using two types of commercial assay. Women and children were followed according to a standard protocol. Adjusted odds ratios (AOR) were calculated to estimate the effect of RNA load and other maternal factors on vertical transmission. RESULTS Maternal RNA levels, mode of delivery and gestational age were independently associated with transmission. Vertical transmission increased with increasing RNA levels, but there was no threshold below which transmission did not occur. The risk was more than double for women with RNA above the sample specific median [AOR 2.36 (1.23-4.52)]. Elective caesarean section was associated with a substantial and significant decrease in transmission [AOR 0.19 (0.06-0.55)], and delivery before 37 weeks gestation with an increased risk [AOR 2.67 (1.33-5.38)]. Elective caesarean section was effective in both subgroups defined by median RNA level [AORs 0.37 (0.08-1.71) and 0.15 (0.03-0.64) below and above median respectively]. The predicted rate of transmission in a woman with a low RNA load delivering by elective caesarean section or vaginally after 37 weeks is around 2%, and 11%, respectively. INTERPRETATION Mother-to-child transmission of HIV-1 is multi-factorial; high RNA load is an important determinant but clearly not the only one. Interventions that target risk factors other than maternal RNA load remain important.

Increased expression of epidermal fatty acid binding protein, cofilin, and 14‐3‐3‐σ (stratifin) detected by two‐dimensional gel electrophoresis, mass spectrometry and microsequencing of drug‐resistant human adenocarcinoma of the pancreas

Abstract: In order to study possible mechanisms leading to chemoresistance in pancreatic adenocarcinoma we examined the global protein expression of pancreatic cancer cells in vitro. We used a cell culture model derived from the adenocarcinoma of the pancreas (EPP85-181P). A classical multidrug-resistant subline, EPP85-181RDB, selected in presence of daunorubicin, and an atypical multidrug-resistant cell variant, EPP85-181RNOV, selected in presence of mitoxantrone, were analyzed using two-dimensional electrophoresis. After staining and image analysis, spots of interest were isolated using preparative two-dimensional electrophoresis and subjected to mass spectrometry and microsequencing. Three proteins, E-FABP, cofilin, and 14-3-3-sigma (stratifin), were overexpressed in chemoresistant cell lines. Cofilin was present in both multidrug in chemoresistant cell lines. Cofilin was present in both multidrug-resistant cell lines. E-FABP and 14-3-3-sigma (stratifin) was found to be overexpressed only in the mitoxantrone-selected atypical multidrug-resistant cell line. The possible significance of these findings is discussed.

Remineralisation und Schmerzlinderung von Knochenmetastasen nach unterschiedlich fraktionierter Strahlentherapie (10mal 3 Gy vs. 1mal 8 Gy) Eine prospektive Studie

Abstract: Es wurde eine prospektive randomisierte Studie zur Bestrahlung von Knochenmetastasen durchgefuhrt, die sowohl die Schmerzansprechrate als auch die Remineralisation als Kriterien des Therapieerfolgs berucksichtigte. Patienten und Methode: 107 Patienten mit histologisch gesichertem Mamma-, Bronchial-, Prostata- und Nierenzellkarzinom, die eine schmerzhafte und rontgenologisch sichtbare Osteolyse hatten, wurden in die Studie aufgenommen. Es erfolgten eine Stratifizierung nach den Entitaten und eine Randomisation in zwei unterschiedliche Therapiearme (10mal 3 Gy vs 1mal 8 Gy). Die Erfassung der Schmerzansprechrate erfolgte mit Hilfe eines Schmerz- und Schmerzmittel-Scores und der subjektiven Einschatzung der Patienten. Die Remineralisation wurde am Computertomographen gemessen. Sowohl die Schmerzreduktion als auch die Remineralisation wurden vor, unmittelbar nach der Therapie sowie sechs Wochen, drei und sechs Monate nach Bestrahlungsabschlus erfast. Ergebnisse: Bei der Schmerzreduktion ergab sich im Vergleich beider Therapiearme sowohl hinsichtlich der Gesamtansprechrate (81% vs. 78%) als auch der kompletten Schmerzreduktion (33% vs. 31%) kein Unterschied. Im 1mal-8-Gy-Arm wurde ein etwas rascheres Ansprechen auf die Therapie beobachtet. Bei der Remineralisation ergab sich ein hochsignifikanter Unterschied (p < 0,0001) im Vergleich aller Patienten zugunsten des fraktionierten Therapiearms (173% vs. 120%). Im fraktionierten Arm zeigte sich ein signifikanter Unterschied zwischen Mamma- und Bronchialkarzinomen (p = 0,015). Dieser Unterschied wurde im Vergleichsarm (1mal 8 Gy) nicht gesehen. Beim Vergleich der unterschiedlichen Therapiearme hinsichtlich der einzelnen Tumorentitaten zeigte sich zwar bei allen eine Tendenz zugunsten des fraktionierten Therapiearmes, ein signifkanter Unterschied wurde nur beim Mammakarzinom gemessen (p < 0,001). Schlusfolgerung: Schmerzreduktion und Remineralisation liegen unterschiedliche Wirkungsmechanismen zugrunde. Bei alleiniger Berucksichtigung der Schmerzen ist ein Kurzzeitschema zu empfehlen, da es effektiv und wenig belastend fur den Patienten ist. Unter Berucksichtigung der Remineralisation ist einem fraktionierten Schema der Vorzug zu geben, da hiermit eine grosere biologische Wirksamkeit erzielt wird und damit die Grundlage fur eine bessere Stabilisierung gegeben ist.

Developmental expression of sodium entry pathways in rat nephron.

Abstract: During the past several years, sites of expression of ion transport proteins in tubules from adult kidneys have been described and correlated with functional properties. Less information is availab...

Alcohol Withdrawal in the Surgical Patient: Prevention and Treatment

Abstract: In the literature on AWS, there is repeated emphasis on performing a thorough preanesthesia assessment in patients with suspected chronic alcohol use. Because these patients are difficult to diagnose and to treat in surgical settings if complications arise, a multimodal approach is highly recommended (86). Ideally, AWS should be prevented by adequate prophylaxis. If AWS develops after surgery or trauma, immediate therapy is required. The symptoms of AWS can be controlled using the combination of a benzodiazepine (in Europe, also chlormethiazole) with haloperidol or clonidine. The drug regimens must be individualized and symptom-oriented to treat hallucinations and autonomic signs. Dosages are generally larger than those in detoxification units. Other approaches to modulate the neuroendocrine-immune axis in patients with an increased risk of postoperative infectious complications look promising but await controlled trials.

No benefit of long-term ciprofloxacin treatment in patients with reactive arthritis and undifferentiated oligoarthritis: a three-month, multicenter, double-blind, randomized, placebo-controlled study.

Abstract: Objective To investigate the effect of long-term antibiotic treatment in patients with reactive arthritis (ReA) and undifferentiated oligoarthritis. Methods One hundred twenty-six patients were treated with ciprofloxacin (500 mg twice a day) or placebo for 3 months, in a double-blind, randomized study. Of these patients, 104 (48 treated with ciprofloxacin and 56 treated with placebo) were valid for clinical evaluation: 55 were diagnosed as having ReA with a preceding symptomatic urogenic or enteric infection and 49 as having undifferentiated oligoarthritis. These 2 groups were randomized separately. The triggering bacterium was sought by serology and/or culture. The percentage of patients in remission after 3 months of treatment was chosen as the primary efficacy parameter. Results A triggering bacterium could be identified in 52 patients (50%): Chlamydiatrachomatis in 13, Yersinia in 14, and Salmonella in 25. No patient was positive for Campylobacterjejuni or for Shigella. No difference in outcome was found between treatment with ciprofloxacin or placebo in the whole group or in subgroups of patients with ReA or undifferentiated oligoarthritis. No difference was seen in patients with a disease duration <3 months. Ciprofloxacin was not effective in Yersinia- or Salmonella-induced arthritis but seemed to be better than placebo in Chlamydia-induced arthritis. This difference was not significant, however, which might be due to the small sample size. Conclusion Long-term treatment of ReA with ciprofloxacin is not effective; however, it might be useful in the subgroup of patients who have Chlamydia-induced arthritis. This has to be proven in a bigger study focusing on patients with Chlamydia-induced arthritis.

Comparison of four magnetic resonance methods for mapping small temperature changes.

Abstract: Non-invasive detection of small temperature changes (C) is pivotal to the further advance of regional hyperthermia as a treatment modality for deep-seated tumours. Magnetic resonance (MR) thermography methods are considered to be a promising approach. Four methods exploiting temperature-dependent parameters were evaluated in phantom experiments. The investigated temperature indicators were spin-lattice relaxation time , diffusion coefficient D, shift of water proton resonance frequency (water PRF) and resonance frequency shift of the methoxy group of the praseodymium complex (Pr probe). The respective pulse sequences employed to detect temperature-dependent signal changes were the multiple readout single inversion recovery (T One by Multiple Read Out Pulses; TOMROP), the pulsed gradient spin echo (PGSE), the fast low-angle shot (FLASH) with phase difference reconstruction, and the classical chemical shift imaging (CSI). Applying these sequences, experiments were performed in two separate and consecutive steps. In the first step, calibration curves were recorded for all four methods. In the second step, applying these calibration data, maps of temperature changes were generated and verified. With the equal total acquisition time of approximately 4 min for all four methods, the uncertainties of temperature changes derived from the calibration curves were less than C (Pr probe C, water PRF C, C and C). The corresponding maps of temperature changes exhibited slightly higher errors but still in the range or less than C (C, C, C, C respectively). The calibration results indicate the Pr probe method to be most sensitive and accurate. However, this advantage could only be partially transferred to the thermographic maps because of the coarse matrix of the classical CSI sequence. Therefore, at present the water PRF method appears to be most suitable for MR monitoring of small temperature changes during hyperthermia treatment.

Phase ii study of gemcitabine as first-line chemotherapy in patients with advanced or metastatic breast cancer

Abstract: In this phase II study, the efficacy and tolerability of gemcitabine were studied in 42 patients with locally advanced or metastatic breast cancer who had received up to one prior chemotherapy regimen in an adjuvant setting. Ten patients had received adjuvant chemotherapy. Twenty-eight patients (67%) had visceral disease spread at entry. Gemcitabine (1000 mg/m2) was administered weekly on days 1, 8 and 15 of a 28-day cycle. The mean number of completed cycles was 3.9 and the mean dose delivered was 942.2 mg/m2. Ninety-seven percent of injections were administered as assigned. No complete responses were observed, but there were six partial responses and 24 patients with stable disease lasting 2-9 months. The overall response rate was 14.3% (95% CI 5.4-28.5%). The median survival for all patients was 15.2 months. Maximum WHO grade 3 and 4 toxicities were observed in five patients for nausea and vomiting, one patient for diarrhea, one patient for pain, seven patients for alanine transaminase, and eight patients for segmented neutrophils. There were no grade 3 and 4 toxicities for alopecia. These data confirm modest single-agent gemcitabine activity in advanced or metastatic breast cancer. Gemcitabine's favorable toxicity profile makes it an ideal candidate for combination chemotherapy.

Potent Antitumor Activity of 2-Methoxyestradiol in Human Pancreatic Cancer Cell Lines

Abstract: We examined the effect of 2-methoxyestradiol (2-ME) on the growth and tumorigenesis of human pancreatic cancer cells. 2-ME inhibited the growth of these cell lines (50-90%) in a dose- and time-dependent fashion, and terminal deoxynucleotidyl transferase staining showed that it induced apoptotic cell death. Flow cytometric analysis indicated that 2-ME-sensitive cells showed a prolonged S phase after 48 h of treatment. We used a mouse model for in vivo studies of lung metastasis and injected MIA PaCa-2 cells into the tail veins of nu/nu mice; lung colonies were formed. Mice given oral 2-ME showed 60% inhibition in the number of lung colonies compared with control, untreated animals. These results suggest that 2-ME may have clinical application for the treatment of pancreatic cancer.