bluuebunny/crossref_metadata_2025 huggingface dataset:
Nephrotoxicity and the Proximal Tubule
Authors: Thévenod Frank
Published: May 2003

Cadmium (Cd2+) is a non-essential heavy metal, which is taken up from the environment into the body through pulmonary and enteral pathways. The S1 segment of the kidney proximal tubule (PT) is a major target of chronic Cd2+ toxicity. Renal dysfunction develops in up to 7% of the general population and in its most severe form displays major features of Fanconi syndrome, such as a defective protein, amino acid, glucose, bicarbonate and phosphate reabsorption. The major pathway for Cd2+ uptake by PT cells (PTCs) in vivo is apical endocytosis of Cd2+ complexed to the high-affinity metal-binding protein metallothionein (MT), which may be receptor-mediated. MT is subsequently degraded in endo-lysosomes, and Cd2+ is liberated for translocation into the cytosolic compartment, possibly using transporters for Fe2+, Zn2+ or Cu2+, such as the divalent metal transporter DMT1. Free Cd2+ ions in the extracellular space are translocated across apical and/or basolateral PTC membranes into the cytosol via transporters, whose identity remains unknown. Cytosolic Cd2+ generates reactive oxygen species (ROS), which deplete endogenous radical scavengers. ROS also damage a variety of transport proteins, including the Na+/K+-ATPase, which are subsequently degraded by the proteasome and endo-lysosomal proteases. Cd2+ causes mitochondrial swelling and release of cytochrome c. If these ROS-mediated stress events are not balanced by repair processes, affected cells undergo apoptosis. But Cd2+ also induces the upregulation of cytoprotective stress and metal-scavenging proteins, such as MT. In addition, Cd2+ upregulates the detoxifying pump multidrug resistance P-glycoprotein, which appears to protect PTCs against Cd2+-induced apoptosis. Thus, Cd2+ interferes with various cellular events ranging from mechanisms of induction of programmed cell death to activation of cell survival genes. A better understanding of the cellular mechanisms involved in Cd2+ nephrotoxicity should provide insights into other heavy metal (e.g. Pb2+, Hg2+) nephropathies and various forms of acquired Fanconi syndrome.
Reversing Transmembrane Electron Flow: The DsbD and DsbB Protein Families
Authors: Kimball Richard A. , Martin Laetitia , Saier Jr. Milton H.
Published: May 2003

DsbD and DsbB are two proteins that in Escherichia coli catalyze transmembrane electron flow in opposite directions, thereby allowing reversible oxidoreduction of periplasmic dithiol/disulfide-containing proteins. We have identified all recognizable homologues of these two proteins in the databases and have conducted structural and phylogenetic analyses of the two families. The larger DsbD family is more diverse in sequence, topology, function and organismal distribution than the smaller DsbB family. DsbB homologues are rarely found outside of the proteobacteria, although DsbD homologues are found in many bacterial kingdoms as well as archaea and plant chloroplasts. Few organisms with a fully sequenced genome and a DsbB homologue lack a DsbD homologue, and most of these DsbD homologues fall within two clusters in the DsbD tree, exhibiting phylogenetic relationships that are the same as those observed for the DsbB proteins. These observations suggest that a subset of the DsbD homologues evolved in parallel with the DsbB family to perform a single unified function involving reversible extracytoplasmic protein dithiol-disulfide bond interchange. DsbD family proteins are shown to have arisen by an internal gene duplication event, and this observation leads to prediction of the pathway taken for the evolutionary appearance of the different protein topological types found within this family.
Phylogenomic Analysis of the Giardia intestinalis Transcarboxylase Reveals Multiple Instances of Domain Fusion and Fission in the Evolution of Biotin-Dependent Enzymes
Authors: King Jordan I. , Henze Katrin , Fedorova Natalie D. , Koonin Eugene V. , Galperin Michael Y.
Published: May 2003

Sequencing of the gene encoding a pyruvate carboxylase-like protein from the amitochondrial eukaryote Giardiaintestinalis revealed a 1,338 aa protein composed of acetyl-CoA carboxyltransferase (ACCT), pyruvate carboxyltransferase (PycB), and biotin carboxyl carrier protein (BCCP) domains, linked in a single polypeptide chain. This particular domain combination has been previously seen only in the methylmalonyl-CoA:pyruvate transcarboxylase from Propionibacteriumfreudenreichii, where each of these domains is encoded by an individual gene and forms a separate subunit. To get an insight into the evolutionary origin and biochemical function of the G.intestinalis enzyme, we compared its domain composition to those of other biotin-dependent enzymes and performed a phylogenetic analysis of each of its domains. The results obtained indicate that: (1) evolution of the BCCP domain included several domain fusion events, leading to the ACCT-BCCP and PycB-BCCP domain combinations; (2) fusions of the PycB and BCCP domains in pyruvate carboxylases and oxaloacetate decarboxylases occurred on several independent occasions in different prokaryotic lineages, probably due to selective pressure towards co-expression of these genes, and (3) because newly sequenced biotin-dependent enzymes are often misannotated in sequence databases, their annotation as either carboxylases, decarboxylases, or transcarboxylases has to rely on detailed analysis of their domain composition, operon organization of the corresponding genes, gene content in the particular genome, and phylogenetic analysis.
Radiation and Functional Specialization of the Family-3 Glycoside Hydrolases
Authors: Cournoyer B. , Faure D.
Published: May 2003

A phylogenetic analysis of the glycoside hydrolases of family 3 (GH3s) was conducted in order to infer particular trends in its evolution: functional specialization, gene transfer events, gene duplications and paralogous evolution, and gene deletions. The phylogenetic analysis of GH3s revealed six clusters, i.e., A, B, C, D, E, and F that could fit the definition of 3 sub-families, i.e., AB, AB′ and AB′′. While the sub-families AB′ and AB′′ contain a single cluster, F and E, respectively, the AB sub-family is sub-divided into four clusters. Global analysis of the GH3 phylogenetic tree suggests a primary burst of amplification of the GH3s that might have led to these sub-families. Specializations, gene transfers, and gene duplications among each of these sub-families and phylogenetic clusters might then have occurred and have been inferred. The fine comparison of the enzyme properties and phylogenetic relationships of GH3s allowed to detect common functional groups that belong to the same cluster (D, E or F), or sub-cluster (A1, A2 or B2). The prokaryotic and eukaryotic β-xylosidases and β-glucosidases belong to the AB and AB′ sub-families, and the N-acetylglucosaminidases are in sub-family AB′′ (in cluster E). In some instances (B1, B2, C1, C2, and C3), the lack of data and/or the high heterogeneity of the hydrolytic properties did not allow to infer a particular link between an enzyme functional group and a phylogenetic cluster, suggesting the emergence of some highly specialized GH3s.
The Effect of Estradiol in the Striatum Is Blocked by ICI 182,780 but Not Tamoxifen: Pharmacological and Behavioral Evidence
Authors: Xiao Li , Jackson Lisa R. , Becker Jill B.
Published: May 2003

Estradiol in the striatum enhances amphetamine (AMPH)- or KCl-stimulated dopamine (DA) release and the pacing of sexual behavior in the female rat. These effects of estradiol in the striatum are rapid, steroid specific and thought to be mediated by a G protein-coupled membrane receptor for estradiol. In the current experiments, we examined whether two antiestrogens, ICI 182,780 (ICI) and tamoxifen (TAM), affect the enhancement by estradiol of (1) AMPH-induced DA release from striatal tissue in vitro, and (2) paced mating behavior in the female rat. The steroidal antiestrogen ICI significantly blocked the effect of estradiol on AMPH-induced striatal DA release. In contrast, the nonsteroidal, triphenylethylene antiestrogen TAM did not block the effect of estradiol, even when a concentration 10 times the concentration of estradiol was delivered to the tissue. Neither of the compounds showed estrogenic action when tested in the absence of estradiol. When implanted into the dorsolateral striatum, ICI inhibited the effect of estradiol on pacing of sexual behavior, while TAM did not. The specific effects of ICI, but not TAM, on striatal DA release and pacing behavior provide further information about the binding site for estradiol in the striatum.
Regulation of Gonadotropin Releasing Hormone Release by Neuropeptide Y at the Median Eminence during the Preovulatory Period in Ewes
Authors: Advis J.P. , Klein J. , Kuljis R.O. , Sarkar D.K. , McDonald J.M. , Conover C.A.
Published: May 2003

The median eminence (ME) of the hypothalamus is known to be an important brain site where hypophysiotropic release might be regulated by excitatory and inhibitory signals impinging on their neuronal terminals. Since a role for neuropeptide Y (NPY) on preovulatory luteinizing hormone (LH) release has been suggested, we hypothesized that NPY might act at the ME to control preovulatory gonadotropin-releasing hormone (GnRH) release and thus the onset of the preovulatory surge of LH. To examine this possibility, we used the ewe as an animal model to determine: (a) immunocytochemical distribution of GnRH and NPY in the ewe ME; (b) changes in in vivo release of NPY and GnRH using ME push-pull cannula (PPC) perfusate samples, as well as in plasma LH, during the luteal, follicular and preovulatory phases of a synchronized estrous cycle, and (c) effects of ME perfusion of NPY or a Y1-NPY antagonist, or an NPY antiserum on in vivo release of ME-GnRH and plasma LH during a synchronized follicular phase. Immunolocalization reveals a dense plexus of beaded GnRH-containing neurites in the arcuate nucleus and in its vicinity, the pituitary stalk and the palisade. In contrast, a dense plexus of NPY-containing neurites occurs in the internal layer, with occasional fibers found in the intermediate and lateral external zone of the ME. In the area between the lateral internal and lateral external layers, both NPY and GnRH-containing processes were found, thus providing opportunities for synaptic and/or paracrine interactions between NPY- and GnRH-containing neurons. Hormonal analysis indicated that a synchronized preovulatory surge of LH is elicited within a 2-hour window by the sequential implantation and removal of silastic-encased estradiol (E2) or progesterone (P4) implants. In this paradigm, there was a parallel increase in ME release of both NPY and GnRH preceding the synchronized LH surge. The onset of this synchronized LH surge was advanced by ME perfusion of exogenous NPY and was both delayed and blunted by ME perfusion with the NPY antagonist (both were perfused through the PPC probe for 2 h, starting 2–3 h before the expected onset of the LH surge). In addition, NPY perfusion in the ME increases, while perfusion of the Y1-NPY antagonist or of the NPY antiserum decreases ME-PPC GnRH content and plasma levels of LH in early follicular ewes. Finally, perfusion of NPY antiserum during an ongoing LH surge disrupted LH release. These results suggest that interactions between NPY and GnRH neurons are important in controlling the timing, magnitude and maintenance of the preovulatory LH surge.
Estradiol and Testosterone Have Opposite Effects on Microtubule Polymerization
Authors: Kipp Jingjing Liu , Ramirez Victor D.
Published: May 2003

We have reported earlier the purification of tubulin from a plasmalemmal-microsomal fraction derived from rat hippocampus using an estradiol (E2) affinity column and the specific binding of tubulin to both E2 and testosterone (T). To further investigate the effect of E2 and T on the function of this protein, changes in microtubule polymerization as a result of exposure to the steroids were examined in this study, using both pure tubulin and rat hippocampal primary cell cultures. First, pure tubulin was incubated with or without steroids for 30 min on ice followed by polymerization at 37°C. The numbers of microtubules formed were counted from electron microscopic pictures. The results showed that at 30 min of polymerization, 10 nM, 30 nM and 30 µM of E2 inhibited microtubule assembly by –70%, –94%, and –92%, respectively (p < 0.01), while T at the same three concentrations stimulated it by +83%, +66%, and +121%, respectively (p < 0.05). The inhibitory effect of E2 and the stimulatory effect of T were observed at 15, 30 and 60 min of the polymerization process. Next, primary cell cultures from 17-day rat fetus hippocampal tissues were treated with the steroids and polymerized microtubules (Triton X-100 resistant) were examined by immunocytochemistry. The results demonstrated that 60 min of E2 treatment (10 nM) decreased the intensity of the immunolabeling of polymerized microtubules. The effect of T at nM concentration was not significant though it increased the immunolabeling at µM concentration. Of great significance was a remarkable inhibition by T of the well-established depolymerization effect of colchicine in both the pure tubulin assay and the cell culture model, while E2 was not effective. In an effort to pursue the possible mechanism(s) of the effect of E2 and T on microtubule formation, we found that T only inhibited the microtubule depolymerization process without affecting the rate of polymerization. In contrast, E2 modifies only the polymerization process without altering the depolymerization. Overall, these data indicate that E2 and T may be considered as novel regulators of microtubule dynamics and thereby controlling cytoskeleton function in cells.
Phase II Study of Irinotecan and Ifosfamide in Patients with Advanced Non-Small Cell Lung Cancer
Authors: Ichiki Masao , Rikimaru Toru , Gohara Rumi , Koga Takeharu , Kawayama Tomotaka , Matunami Michiya , Oshita Yuichi , Kamimura Tomoko , Aizawa Hisamichi
Published: May 2003

Objectives: This phase II study was conducted to investigate the efficacy and safety of irinotecan (CPT-11) and ifosfamide as first-line chemotherapy for advanced non-small cell lung cancer (NSCLC). Methods: Eligibility criteria included histologically or cytologically confirmed NSCLC (stage IIIb or IV), no prior treatment, and measurable or evaluable disease. CPT-11 (80 mg/m2) was administered intravenously on days 1, 8, and 15, while ifosfamide (1.5 g/m2) was given on days 1 through 3 every 4 weeks. Results: Forty-four patients (31 men) with a median age of 65 years (range 43–75) and a median ECOG performance status of 1 (range 0–2) were enrolled. The response rate was 29.5% [95% CI: 16.7–45.2%], with 13 partial responses. The median survival was 12.5 months, the median time to progression was 5.3 months, and the 1 and 2-year survival rates were 52.3 and 11.3%, respectively. Toxicity was generally mild; WHO grade 3–4 neutropenia was recorded in 38.6% of the patients, grade 3 diarrhea in 6.8%, and grade 3–4 nausea/vomiting in 0%. Conclusions: CPT-11 combined with ifosfamide demonstrated anti-tumor activity in advanced NSCLC, with response and survival rates similar to those of cisplatin-based chemotherapy but with a more favorable toxicity profile.
18F-Fluorodeoxyglucose Positron Emission Tomography (18F-FDG-PET) for Staging and Follow-Up of Marginal Zone B-Cell Lymphoma
Authors: Hoffmann Martha , Kletter Kurt , Becherer Alexander , Jäger Ulrich , Chott Andreas , Raderer Markus
Published: May 2003

Objective: According to recent reports, nodal marginal zone lymphoma (MZL) appears to be a distinctive lymphoma entity rather than a more advanced stage of extranodal MZL of mucosa-associated lymphoid tissue (MALT). We have therefore retrospectively evaluated all patients diagnosed with nodal or extranodal MZL who have been referred to our unit for imaging using 18F-fluorodeoxyglucose positron emission tomography (18F-FDG-PET). Patients and Methods: A total of 21 patients with a diagnosis of MZL upon referral for imaging with 18F-FDG-PET were identified. Histological reassessment of biopsy specimens confirmed the diagnosis of extranodal MZL of MALT in 14 patients, while a diagnosis of nodal MZL was verified in 6 patients. Lymphoma cell proliferation was assessed immunohistochemically using a Ki-67 antibody. Whole-body 18F-FDG-PET scans were performed on a GE advanced PET scanner 40 min after intravenous injection of 300–380 MBq 18F-FDG. Results: None of the patients with extranodal MZL showed focal tracer uptake within verified tumor sites. In contrast, 5 of the 6 patients with nodal MZL showed significant FDG uptake within the affected lymph nodes. These results did not simply reflect the different growth fractions of the two lymphoma entities since the proliferation indices of the two groups did not differ significantly. Conclusion:18F-FDG-PET visualizes nodal MZL in a high proportion of patients whereas FDG uptake is undetectable in extranodal MZL. Although limited by the small number of patients, this study suggests that imaging with 18F-FDG-PET might play a potential role in the diagnostic workup of patients with nodal MZL involvement.
Clinicopathological Significance of Nup88 Expression in Patients with Colorectal Cancer
Authors: Emterling Anna , Skoglund Johanna , Arbman Gunnar , Schneider Jose , Evertsson Sofia , Carstensen John , Zhang Hong , Sun Xiao-Feng
Published: May 2003

Objective: The nucleoporin Nup88 is overexpressed in a series of human malignancies, however, its clinicopathological significance has not been studied. Our aims were to analyze Nup88 expression in normal mucosa, primary tumors and metastases from colorectal cancer patients and further to identify relationships of Nup88 expression with clinicopathological and other factors. Materials and Methods: Using immunohistochemistry, we investigated Nup88 expression in 198 primary colorectal tumors, 96 normal mucosa samples and 35 lymph node metastases. Results: The results showed that the intensity of Nup88 expression increased from the normal mucosa to the primary tumors (p < 0.0001) and tended to increase from the primary tumors to the metastases (p = 0.15). Both primary tumors and metastases presented stronger expression in the invasive margin and vascular-invaded areas. Nup88 expression was positively related to distal tumor location (p = 0.01), infiltrative growth pattern (p = 0.04) and higher proliferative activity (p = 0.04) and reversely to the grade of differentiation (p = 0.02) and apoptosis (p = 0.049). Strong expression of Nup88 predicted a worse outcome in the patients with distal tumors during the follow-up period of up to 3 years (p = 0.02). Conclusions: It seems that overexpression of Nup88 was involved in the tumorigenesis and aggressiveness of colorectal cancers, and Nup88 may be used as a prognostic factor in patients with distal tumors.
Characteristic Cytokine Generation Patterns in Cancer Cells and Infiltrating Lymphocytes in Oral Squamous Cell Carcinomas and the Influence of Chemoradiation Combined with Immunotherapy on These Patterns
Authors: Yamamoto Tetsuya , Kimura Tsuyoshi , Ueta Eisaku , Tatemoto Yukihiro , Osaki Tokio
Published: May 2003

Objectives: Cytokines produced by tumor cells and tumor-infiltrating lymphocytes (TIL) appear to regulate tumor cell growth and the cytotoxic activity of TIL. The objectives of the present study were to investigate cytokine generation patterns in tumor cells and TIL and to examine the influence of cancer therapy on this cytokine production and the cytotoxic activity of TIL. Methods: We determined the levels of cytokines produced by tumor cells and TIL in vitro and measured the cytotoxic activity of TIL against Daudi cells in patients with oral squamous cell carcinoma (OSC) before and 1 week after the start of concomitant chemo-radio-immunotherapy. Results: Before the therapy, OSC cells generated higher levels of granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-α (TNF-α) and transforming growth factor-β (TGF-β) than did oral keratinocytes isolated from the noninflamed gingivae of healthy individuals, but both kinds of cells generated similar levels of interleukin (IL)-1β and IL-6. Compared with peripheral blood mononuclear cells (PBMC) of the patients, TIL produced higher levels of IL-1β, IL-6, IL-10, TNF-α and TGF-β, whereas their production of IL-12 and interferon-γ (IFN-γ) was only slightly higher than that in PBMC. After 1 week of therapy, the cytokine production by OSC cells had largely decreased, while the production of TNF-α, IFN-γ, TGF-β and IL-12 by TIL had increased greatly, although other cytokine levels were almost constant during the investigations. The cytotoxic activity of TIL was higher than that of PBMC before the therapy, and this activity was strongly increased by 1 week of therapy. Conclusions: These results suggest that the cytokine productivities of TIL and tumor cells differ from those of PBMC and normal keratinocytes, respectively, and that chemo-radio-immunotherapy modulates in situ cytokine generation, which is advantageous for inhibition of tumor cell growth and activation of TIL.
Sensitive Detection of Human Telomerase Reverse Transcriptase mRNA in the Serum of Patients with Hepatocellular Carcinoma
Authors: Miura N. , Shiota G. , Nakagawa T. , Maeda Y. , Sano A. , Marumoto A. , Kishimoto Y. , Murawaki Y. , Hasegawa J.
Published: May 2003

Background/Aim: Telomerase reverse transcriptase protein (hTERT) mRNA has been reported to be detectable by reverse transcription polymerase chain reaction (RT-PCR) in the serum of patients with breast cancer. We measured serum hTERT mRNA in patients with hepatocellular carcinoma (HCC), and examined its clinical usefulness. Methods: We performed RT-PCR to detect the expression of hTERT mRNA in 78 patients with HCC, 10 with liver cirrhosis (LC), 12 with chronic hepatitis (CH), and 34 healthy individuals without any liver diseases and cancers, and statistically analyzed the association with clinical parameters which include age, sex, etiology, Child classification, underlying liver disease, biochemical data, α-fetoprotein (α-AFP) number and size of tumor, and histological differentiation of HCC regarding HCC patients. Results: 70 of 78 (89.7%) patients with HCC, 7 of 10 (70.0%) with LC, and 5 of 12 (41.7%) with CH were positive for hTERT expression, whereas all healthy individuals were negative for it. A multivariate analysis showed that positivity of hTERT mRNA was independently associated with AFP, tumor size, and differentiation degree. Conclusions: These results suggest that this assay is sensitive enough to detect hTERT mRNA in serum, and that it would be applicable for early detection and diagnosis of HCC or other cancers by a quantitative method.
Expression of Cancer/Testis Tumor Associated Antigens in Cervical Squamous Cell Carcinoma
Authors: Sarcevic Bozena , Spagnoli Giulio C. , Terracciano Luigi , Schultz-Thater Elke , Heberer Michael , Gamulin Marija , Krajina Zdenko , Oresic Tomislav , Separovic Robert , Juretic Antonio
Published: May 2003

We investigated the expression of tumor-associated antigens (TAA) of the cancer/testis (C/T) gene family in cervical squamous cell carcinomas. First, we focused on the HeLa cervical cancer derived cell line, and we found that it expresses MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A12, GAGE-3/6, LAGE-1, and PRAME genes, encoding defined C/T TAA. In contrast, no expression of MAGE-A10, BAGE, GAGE-1/2, or NY-ESO-1 genes was observed. Corresponding gene products could also be detected by immunoblotting and immunocytochemistry, taking advantage of monoclonal antibodies recognizing discrete TAA. Capitalizing on these data, a monoclonal antibody predominantly recognizing MAGE-A4 TAA in paraffin-embedded sections (57B) was used to investigate the C/T gene expression in clinical tumor samples. A group of 60 patients was studied, and 57B positivity was detectable to different extents in 33% of the cases (20/60). In 13 of them (21%), staining of over 50% of the tumor cells was evident, whereas healthy cells always scored negative. Remarkably, MAGE-A4 expression was significantly (p < 0.05) more frequently detectable in poorly differentiated tumors (8/13) than in well-differentiated or moderately differentiated cancers (3/15 and 9/32, respectively) and in stage FIGO II as compared with stage FIGO Ib tumors (12/23 and 5/24, respectively, p = 0.04). Interestingly, staining was mostly nuclear in well-differentiated tumors, but involved both nuclei and cytoplasm in less differentiated cancers. Positivities of comparable frequency were also detectable in a smaller series of specimens upon staining with MAGE-A1- or NY-ESO-1/LAGE-1-specific reagents. Considering the high tumor specificity of C/T TAA, our data provide the rationale for the design of immunotherapy procedures targeting these antigens in cervical cancers.
Brefeldin A Induces Apoptosis and Cell Cycle Blockade in Glioblastoma Cell Lines
Authors: Pommepuy Isabelle , Terro Faraj , Petit Barbara , Trimoreau Frank , Bellet Virginie , Robert Sandrine , Hugon Jacques , Labrousse François , Yardin Catherine
Published: May 2003

Brefeldin A (BFA), a fungal metabolite known to affect the structure and function of the Golgi apparatus, has recently been shown to induce apoptosis and cell growth inhibition in various human cell lines. Glioblastomas (GB) are cerebral tumors with poor prognosis, which display resistance to current therapies including radio- and chemotherapy. The objective of this study was to investigate BFA effects in three human GB cell lines (SA4, SA146 and U87MG cells). Compared with control cells, about 60% of cell growth inhibition was observed in BFA (100 ng/ml for 24 h)-exposed cells in the three cell lines. Furthermore, in SA4 and SA146 cells, BFA was able to induce a time- and dose-dependent apoptosis detected by DAPI staining, TUNEL assay and flow-cytometric analysis. Since p53 expression was not modified after BFA exposure, BFA-induced apoptosis may follow a p53-independent pathway, as already reported. In the same way, BFA did not alter Bcl-2, Bax and Mcl-1 expression. Cell cycle analysis revealed a cell cycle arrest in early G0/G1 phase with an increase in G0/G1 cell population (70% in control cells vs. 83% in exposed cells) associated with a decrease in the S cell population (14% in control cells vs. 5.5% in exposed cells). The Ki67 labeling index also confirmed the cell cycle blockade. Our results suggest that BFA may be a potent cell cycle modulator and inducer of apoptosis in GB cell lines, and therefore may become a promising candidate for the chemotherapeutic treatment of gliomas.
Gender Differences in Dissociation
Authors: Spitzer Carsten , Klauer Thomas , Grabe Hans-Joergen , Lucht Michael , Stieglitz Rolf-Dieter , Schneider Wolfgang , Freyberger Harald J.
Published: May 2003

Considering that epidemiological research on dissociative disorders has suggested a 9 to 1 predominance of female cases, this study investigated the relationship between gender and dissociation using a dimensional approach. A total of 2,153 participants from different diagnostic groups completed the Dissociative Experience Scale. In order to control for the confounding effect of current psychopathology a subgroup 790 subjects additionally completed the SCL-90. We did not find any differences in the general or pathological dissociation scores. Hypothetical gender differences in dissociative psychopathology were not a function of diagnostic categories. There were no significant sex differences in the distribution of high dissociators. Our findings suggest that men and women do not generally differ in dissociative psychopathology. The implications for future investigations on the epidemiology, etiology, and psychobiology of dissociative symptoms are discussed.
Personality Disorders and Self-Perceived Quality of Life in an Elderly Psychiatric Outpatient Population
Authors: Condello Caterina , Padoani Walter , Uguzzoni Ugo , Caon Federico , De Leo Diego
Published: May 2003

The aim of this work was to assess the impact of personality disorder and traits on the subjective assessment of quality of life by a sample of elderly psychiatric outpatients (60 years or more). Sixty of the total number of subjects interviewed were included in the test sample since they met the Vragenlijst Klinishe Persoonlijkheid (VKP) criteria for personality disorder. A control sample was also formed, composed of 50 subjects without personality disorders or traits. Quality of life was assessed using the LEIPAD questionnaire. Personality disorder diagnoses were prevalently distributed in cluster C (25%) and in cluster A (20%). A total of 48.3% of patients (n = 29) had more than 1 personality disorder. Sixty percent of the sample (n = 36) presented with comorbid Axis I psychiatric disorders, in most cases depressive pathologies. Comparing the case and control groups, worse LEIPAD scores were achieved by subjects with personality disorders. Between clusters of personality disorders, the worst scores were observed in subjects with more than 1 personality disorder diagnosis. Elderly people who presented with a personality disorder were more vulnerable to late-life distress, especially those with more than one diagnosis. In keeping with linear regression analysis, widowhood, divorcehood and living with others appear to be variables able to negatively influence quality of life in this population. While the VKP needs to be adapted to the application in the elderly, it does seem to be a particularly useful screening instrument.
Endometrial Receptivity: Selective Adhesion Competence of Rabbit Uterine Epithelium for Trophoblast but Not for Various Tumor Cells
Authors: Hohn H.-P. , Donner A.J. , Denker H.-W.
Published: May 2003

A modification of an established in vitro model for embryo implantation was used to probe the receptive uterine epithelium for any specificity of interaction with various invasive cells other than trophoblast. Endometrial explants consisting of stroma and epithelium taken from pseudopregnant rabbits were cultured in the presence of progesterone in order to regenerate a complete epithelial lining while maintaining the receptive state. Such precultured fragments were brought into contact with multicellular spheroids of different invasive tumor cell lines from different species. In contrast to the trophoblast of the rabbit blastocyst (previous publication), none of the tumor cell lines was able to adhere to intact epithelium of endometrial fragments nor to penetrate it. The uterine epithelium was also an insurmountable barrier for tumor cell spheroids confronted with the epithelium of fresh complex explants consisting of endometrium and myometrium or for spheroids introduced into the uterine lumen of pregnant/pseudopregnant rabbits at the periimplantation phase. However, all tumor cells were able to adhere to and mostly also to invade into the endometrial stroma when it was exposed artificially, i.e. when the epithelium was removed. These results suggest that the receptivity of rabbit uterine epithelium shows a remarkable selectivity with respect to cell type (trophoblast) and species (rabbit, not human, mouse, or rat).
Embryonic Blastemic Changes in Retinoic Acid-Induced Hindlimb Deformity
Authors: Santos-Alvarez Inmaculada , Martos-Rodríguez Antonia , Delgado-Baeza Emilio
Published: May 2003

In this study we analyzed, on an experimental model, blastemic changes occurring during the embryonic period that may later cause fetal hindlimb deformity. Experimental induction of clubfoot-like deformity in rat fetuses was performed by maternal administration of retinoic acid (RA) (120 mg/kg body weight) as a single intragastric dose on day 10 of pregnancy (previous phase of this assay). The caudal and hindlimb blastemic changes were studied by mitosis count and stereological, immunohistochemistry and AgNOR techniques in the hindlimbs of 15-day embryos and in the caudal somites of 11-day embryos. In 15-day embryos from the assay group, hypoplasia and misorientation of hindlimbs were present in 90% of the cases. The histological study showed a blastemal defect as follows: (a) reduction in mesenchymal cell activity (mitotic and AgNOR activities); (b) increase of volume of vascular lumen; (c) reduction in volume of nerve structures, and (d) reduction in the percentage of pre-rhabdomyoblastic cells. In 11-day embryos from the assay group, caudal somites showed disruption, including loss of usual morphology. Moreover, somitic AgNOR activity decreased compared to the control group. The greatest reduction in the number of black-dots per cell was in the myotome. These findings suggest that a certain pathology of the somites, very little studied to date, might be involved in clubfoot pathogenesis.
Role of Adjuvant Chemoradiotherapy for Abdominal Malignancies
Authors: Glimelius Bengt
Published: May 2003

The rationale for combining surgery, radiotherapy and chemotherapy is discussed and the clinical results seen with surgery and adjuvant radiochemotherapy in three major abdominal malignancies are reviewed. A systematic approach to the literature was used. In rectal cancer, postoperative radiochemotherapy is an established treatment, although there is weak scientific support for the combined approach. The same clinical gains can also be reached much more easily with preoperative radiotherapy. In gastric cancer, a recent large randomised trial showed improved survival from postoperative radiotherapy. This was not seen in a comparably large trial in pancreatic cancer. The reasons for the different results according to primary tumour site are discussed. It is argued that adequate coverage of all adjacent regional lymph node stations is necessary for an effect on survival.
A New Blade for the Bookwalter Retractor System
Authors: Wang Shyh-Jen , Wu Chew-Wun
Published: May 2003

The table-fixed Bookwalter retractor system has been widely adopted in abdominal surgery, and during the past 15 or more years, we have succeeded in using the retractor system for thousands of cases operated on at the Taipei Veterans General Hospital, Taiwan. However, for some surgical procedures, such as gastrectomy, the blades that are currently available do not adequately hold deeper organs or tissue, such as the liver. We have successfully designed and fabricated a blade for grasping deeper tissue or organs, which is reusable and cost-effective. The shape of this new blade is described.
Mutations in Exon 11 of the c-kit Gene in a Myogenic Tumor and a Neurogenic Tumor as Well as in Gastrointestinal Stromal Tumors
Authors: Yasuoka Rie , Sakakura Chohei , Shimomura Katsumi , Fujita Yoshifumi , Nakanishi Masayoshi , Aragane Hideki , Hagiwara Akeo , Bamba Masamichi , Abe Tatsuo , Yamagishi Hisakazu
Published: May 2003

Background/Aims: Gain-of-function mutations in exons 9, 11 and 13 of the c-kit gene in gastrointestinal stromal tumors (GISTs) have been identified, and it has been reported that the prognosis is worse for patients with mutation-positive GISTs than for those with mutation-negative GISTs. We studied c-kit mutations in gastrointestinal mesenchymal tumors. By chance, the c-kit mutation in exon 11 was found in myogenic and neurogenic tumors as well as in GISTs. Furthermore, we studied the clinical prognostic utility of these mutations. Methods: Ten gastrointestinal mesenchymal tumors were stained with HE and immunohistochemically analyzed with α-smooth muscle actin, S-100 protein, CD34 and c-kit. In these tumors, as well as in 11 cases of leiomyomas, PCR-amplified DNA from the juxtamembrane (JM) domain of exon 11, the extracellular domain of exon 9 and the tyrosine kinase domain 1 of exon 13 showed a high frequency of c-kit mutation and was sequenced. Results: Although c-kit mutations have previously been reported only in GISTs, we found c-kit mutations in the JM domain of exon 11 in one myogenic and one neurogenic tumor as well as in two GISTs. No c-kit mutation was seen in the 11 cases of leiomyomas. In addition, all four cases with c-kit mutation in exon 11 suffered a relapse sooner than the other cases without c-kit mutations. Conclusion: Clinically, the prognosis was worse for the patients with mutation-positive gastrointestinal mesenchymal tumors than for those with mutation-negative tumors. We therefore conclude that the gain-of-function mutation in exon 11 of the c-kit gene is an important prognostic factor for gastrointestinal mesenchymal tumors, including myogenic and neurogenic tumors as well as GISTs.
Serum Concentration of Type IV Collagen 7S Domain as a Marker for Increased Risk of Recurrence after Liver Resection for Hepatocellular Carcinoma
Authors: Kawai Shuichi , Kubo Shoji , Tsukamoto Tadashi , Tanaka Hiromu , Shuto Taichi , Yamamoto Takatsugu , Hirohashi Kazuhiro , Kinoshita Hiroaki
Published: May 2003

Background/Aims: Serum concentration of type IV collagen 7S domain (7S collagen) is a marker of hepatic fibrosis. We investigated the usefulness of measuring the serum 7S collagen concentration as a risk factor for recurrence after liver resection for hepatocellular carcinoma (HCC). Methods: The serum 7S collagen concentration was measured before liver resection for HCC in 219 patients. Group 1 included 102 patients with a high serum concentration of 7S collagen (≧8.0 ng/ml), and group 2 consisted of 117 patients with a low serum concentration of 7S collagen (<8.0 ng/ml). Clinicopathologic findings and outcome after surgery were compared between groups. Results: The results of liver function tests were better in group 2 than in group 1. The percentage of patients with cirrhosis or who underwent minor resection was higher in group 1 than in group 2. The percentage of patients with moderately or poorly differentiated HCC or portal invasion was higher in group 2 than in group 1. Univariate and multivariate analyses identified that a high serum concentration of 7S collagen (≧8.0 ng/ml) was an independent risk factor for recurrence. Conclusion: Measuring the serum 7S collagen concentration is useful to estimate the risk of recurrence after resection of HCC.
Spontaneous Endometriosis of the Abdominal Wall
Authors: Ideyi Steve C. , Schein Moshe , Niazi Massoma , Gerst Paul H.
Published: May 2003

Background: The abdominal wall is the commonest site of extrapelvic endometriosis, which usually develops in association with a prior surgical scar. Case: A 28-year-old woman with two endometriomas in the recti abdominis, which developed in a scarless abdomen. Conclusions: Abdominal wall endometrioma can develop spontaneously in the absence of prior surgical scar and should be considered in the differential diagnosis of abdominal wall masses.
The Influence of Surgical Operative Experience on the Duration of First Ventriculoperitoneal Shunt Function and Infection
Authors: Cochrane D.D. , Kestle J.R.W.
Published: May 2003

The relationship of surgeon experience, measured by operative volume, to the outcomes of ventricular shunt treatment of hydrocephalus in children is not clear. This paper explores this relationship based on first ventriculoperitoneal shunts (VPS) implanted in English-speaking Canada during the period from April 1989 to March 2001. Three thousand seven hundred and ninety-four first VPS insertions, performed by 254 surgeons, were reviewed. Surgical experience was represented by the number of shunt operations performed during the study period by each surgeon prior to the date of the operation. The 6-month shunt failure risk for less experienced surgeons was 38%, compared to 31% for more experienced surgeons. This difference decreased to 4% at 60 months and 3% at 120 months (p = 0.001). The infection rate for initial shunt insertions was 7% for patients treated by more experienced surgeons and 9.4% for those treated by less experienced surgeons (p = 0.006). A relationship between surgeon experience and shunt outcome that appears to be based on the operative experience that a surgeon brings to a procedure is in keeping with clinical experience. This observation has implications for public policy, service planning and surgical mentorship during the earlier years of a surgeon’s career.
Pineal Tumors and Associated Lesions: The Effect of Ethnicity on Tumor Type and Treatment
Authors: Knierim David S. , Yamada Shokei
Published: May 2003

Pineal region lesions consist of a wide variety of rare tumor types, including deep midline cysts, intrinsic pineal tumors, germ cell tumors and vascular lesions. Advances in microsurgical, endoscopic and stereotactic techniques have helped to lower morbidity and mortality in the care of patients harboring these lesions. Surgery can be the definitive treatment in cysts and benign lesions. This report summarizes the retrospective experience of the authors with 64 pineal region and associated lesions encountered in multiple institutions over the last 20 years. Histology was obtained in 53 out of 64 radiographically apparent lesions. Direct surgical biopsy of solid and vascular tumors in the pineal region enables precise histological assessment of mixed tumors. By avoiding sampling error, precise treatment can be planned. This series, along with previously published data, shows a much higher incidence of intrinsic pineal tumors, glial tumors and nongerminomatous germ cell tumors in series from North America and Europe than in those from Japan and Korea, where germinoma is much more common. We experienced an incidence of 20.4% germinoma out of 49 solid and vascular pineal tumors, while other authors have described incidences of 51.2 and 53.5%, respectively. The fact that histology is more diverse in Western populations leads to a need to have more representative sampling. Early surgical resection combined with diversion of cerebrospinal fluid is effective in the treatment of pineal lesions and seems to be superior to the alternative of treatment based on the diagnostic response to radiation and/or on tumor markers alone.
Intensity of Long-Term Treatment with Warfarin Is Influenced by Seasonal Variations
Authors: Salobir Barbara , Šabovič Mišo , Peternel Polona
Published: May 2003

In a prospective study, which lasted 1 year and was performed on a large cohort of 800 patients treated at an outpatient anticoagulant clinic, we investigated possible seasonal variations in the intensity of the effect of warfarin treatment. For every season – winter, spring, summer and autumn – the mean prothrombin time reported as an international normalised ratio (INR) and the percentages of INR below 2.0 and above 4.0 were calculated. Significant seasonal variations in mean INR measurements were found, with the lowest values in summer and the highest values in autumn (F = 14.2, p < 0.0001). In addition, a trend toward a higher percentage of INR below 2.0 in summer and above 4.0 in autumn was observed. No significant differences were found between older (>65 years) and younger (<65 years) patients, although there was a trend toward more pronounced variations in younger patients. Consideration of seasonal variations might result in more accurate and safe guidance of warfarin treatment.
Isolation and Characterization of Two Complete Ara h 2 Isoforms cDNA
Authors: Chatel Jean-Marc , Bernard Hervé , Orson Frank M.
Published: May 2003

Background: Ara h 2 is a major peanut allergen recognized by IgE in more than 90% of patients. After electrophoretic separation the purified protein exists as a doublet, and sequences of one incomplete cDNA and one genomic clone for this allergen have been reported. Methods: Ara h 2 isoforms were purified and analyzed by mass spectroscopy, and PCR amplification products of Ara h 2 were cloned and sequenced. Results: Mass spectroscopy of purified Ara h 2 clearly identified a molecular doublet of 16,670 and 18,050 Daltons. Amplification of a peanut cDNA library using PCR primer pairs located at the amino- and carboxy-terminus revealed 2 bands separated by 50 base pairs, which we cloned and sequenced. Two types of complete cDNA clones were obtained, Ara h 2.01 and Ara h 2.02. Compared to Ara h 2.01 and the previously reported cDNA sequences, Ara h 2.02 is characterized by a 12 amino acid insertion starting at position 75 that contains a third repeat of the major IgE binding epitope DPYSPS. Conclusion: We demonstrated the molecular and genetic characteristics of two Ara h 2 isoforms, revealing that one, Ara h 2.02, might be the more potent allergen.
Oral Allergy Syndrome to Chicory Associated with Birch Pollen Allergy
Authors: Cadot P. , Kochuyt A.-M. , van Ree R. , Ceuppens J.L.
Published: May 2003

Background: A few cases of IgE-mediated chicory allergy with oral, cutaneous, and/or respiratory symptoms are reported. We present 4 patients with inhalant birch pollen allergy and oral allergy syndrome to chicory. IgE-binding proteins in chicory and cross-reactivity with birch pollen were studied. Methods: Chicory extract was prepared and immunoblotting was used to study IgE reactivity and cross-reactions with birch pollen. Results: The pattern of IgE binding to chicory was variable among the patients, with protein bands recognized at 18, 21, 40, 52 and 71 kD. Bet v 1-like proteins were detected in chicory by monoclonal antibody binding. Chicory-birch pollen cross-reactivity, as studied in 2 patients from whom enough serum was available, could be demonstrated but did not involve the Bet v 1 protein family. In one of these cases, a 51-kD protein of birch pollen was found to be responsible for cross-reactivity. Conclusions: Chicory should be added to the list of foods that can cross-react with birch pollen and cause the birch pollen-associated oral allergy syndrome.
Complement-Mediated Enhancement of HIV-1 Neutralisation by Anti-HLA Antibodies Derived from Polytransfused Patients
Authors: Wilfingseder Doris , Spruth Martin , Ammann Christoph G. , Döpper Susanne , Speth Cornelia , Dierich Manfred P. , Stoiber Heribert
Published: May 2003

We have recently shown that ‘alloimmune sera’ derived from polytransfused patients (PTP sera) are able to recognise and neutralise HIV in vitro. In this study we try to identify the protein(s), which are recognised by the PTP sera and elucidate mechanisms responsible for the neutralising capacity of these sera. The PTP sera allowed immunoprecipitation (IP) of HLA class II molecules on HIV-infected cells. To detect a potential cross-reactivity of alloreactive antibodies (Ab) with the HIV envelope protein gp160 or its subunits gp120/gp41 and HLA proteins, ELISA and FACS analyses were performed. The lack of reactivity of the PTP sera against rsgp160 in ELISA or FACS analysis indicated that recognition of cells was independent of HIV infection. To clarify whether interaction of the PTP sera with target cells has any effect on the infection process, virus neutralisation assays were performed. Inhibition of HIV infection was observed only when virus was pre-incubated with the PTP sera. Complement enhanced neutralisation of HIV-1 significantly. This enhancement was not due to complement-mediated lysis, because pre-incubation of the target cells with PTP sera did not inhibit HIV replication. Therefore, the neutralising effect of the Ab was due to blocking of the viral attachment/fusion process and not to negative signalling after infection. Since steric hindrance is possible only when HLA and gp120/gp41 are in close vicinity, isolation of rafts and IP assays were performed. These experiments revealed that gp120 and MHC class II molecules are indeed co-localised. The close physical association of gp120/gp41 and HLA strongly supports a mechanism for neutralisation of HIV by anti-HLA-Ab based on steric hindrance.