bluuebunny/crossref_metadata_2025 huggingface dataset:
High Incidence of Angina pectoris in Patients Treated with 5-Fluorouracil
Authors: Wacker Annette , Lersch Christian , Scherpinski Ute , Reindl Laurence , Seyfarth Melchior
Published: August 2003

Objective: Angina pectoris, arrhythmic sudden death and myocardial infarction, all these cardiac events have occasionally been reported during 5-fluorouracil (5-FU) chemotherapy. Underlying mechanisms leading to these events are unknown; damage to the myocytes or vasospasms have been discussed. Methods: 102 consecutive and unselected patients were monitored with 12-lead ECG, echocardiography and radionuclide ventriculography prior to the first cycle of 5-FU chemotherapy and 3 months from baseline. Results: 19% of the patients developed reversible symptoms of angina pectoris during treatment which lasted up to 12 h after cessation of the infusion. Most of the 19 patients showed corresponding ECG changes. 6 out of the 19 patients with severe angina pectoris had subsequent coronary angiography. In none of these patients the coronary angiography showed coronary artery disease, but it showed low ventricular function (ejection fraction <50%) in 2 patients. The ejection fraction did not increase over time. Arrhythmias were screened for with Holter monitoring during 5-FU chemotherapy. The frequency of bradycardia and ventricular extrasystoles increased significantly (p < 0.05) during treatment compared to arrhythmias in Holter monitoring 3 months later. Furthermore the Qtc time in the ECG 3 months later was significantly prolonged (p < 0.05) compared to baseline values. Conclusions: The incidence of angina pectoris in patients during 5-FU treatment seems higher than previously suspected. As myocardial ischemia can be fatal, attentiveness to these symptoms and immediate treatment are crucial.
Features of Second Primary Cancer in Patients with Gastric Cancer
Authors: Ikeda Yoichi , Saku Motonori , Kawanaka Hirofumi , Nonaka Michiyasu , Yoshida Kohji
Published: August 2003

Background: In order to improve the prognosis of gastric cancer patients, the timely identification of second primary cancers is considered to be a crucial clinical problem. Methods: We analyzed the clinicopathological data of 2,250 patients with gastric cancer with regard to both synchronous and metachronous second primary cancers. Results: Of 2,250 patients, 95 (4.2%) had a second primary cancer. Both colorectal and lung cancer were frequently detected, followed by cancer in the liver, esophagus and breast. Regarding the time of detection for such second cancers, 65% of colorectal cancers were detected synchronously, while more than 80% of lung cancers were detected metachronously. The prognosis of gastric cancer patients with a second primary cancer was more negatively influenced by a second primary cancer than by a primary gastric cancer. Conclusion: Since gastric cancer patients may develop synchronous and metachronous second cancers in other organs, effective preoperative and postoperative diagnostic modalities both for second primary cancers, as well as for the recurrence of gastric cancer, need to be developed.
Central Nervous System Involvement in Children with Sarcoma
Authors: Postovsky S. , Ash S. , Ramu I.N. , Yaniv Y. , Zaizov R. , Futerman B. , Elhasid R. , Ben Barak A. , Halil A. , Ben Arush M.W.
Published: August 2003

Objectives: To summarize and analyze the experience in CNS involvement (CNSI) in children with sarcomas treated in the above-mentioned institutions. Patients and Methods: From 1990 to 2001, all medical charts were retrospectively reviewed: 19 sarcoma patients (12 boys and 7 girls) were diagnosed with CNSI (4 osteogenic sarcomas, 11 Ewing sarcomas, 2 rhabdomyosarcomas, 1 alveolar soft part sarcoma and 1 mesenchymal chondrosarcoma). Mean age of all patients at the time of initial diagnosis was 14.9 years (range: 4–24 years), mean age at the time when CNSI was diagnosed was 16.9 years (range: 5.5–27 years). Results: The frequency of CNSI among our patients was 6.17%. The following symptoms and signs (sometimes combined) presented: headache (10 patients), nausea and vomiting (6 patients), seizures (11 patients) and focal neurological signs (9 patients). The mean duration of time elapsed since diagnosis of CNSI till death or last follow-up was 5.2 months (SD: ±5.7 months). Four patients received chemotherapy (CT) alone, 8 CT and radiotherapy (RT), 2 RT alone, 3 supportive treatment only, 1 CT and surgery and 1 surgery alone. Sixteen patients died; there was no significant difference in the duration of survival between those who were treated with RT or surgery (mean ± SD: 6.77 ± 6.56 months) and those who received only CT or supportive treatment (mean ± SD: 2.60 ± 2.94 months) (p = 0.07). Brain disease was the main cause of death in all but 1 patient who died 4 days after autologous bone marrow transplantation from uncontrolled sepsis. In 16 patients, CNSI was part of a metastatic disease. Conclusions: Among children with sarcoma, CNSI is encountered in 6.17% of cases. More effective therapy has to be developed in order to improve their outcome.
Quantitative Analysis of Thymidine Phosphorylase and Dihydropyrimidine Dehydrogenase in Renal Cell Carcinoma
Authors: Morita Tatsuo , Matsuzaki Atsushi , Tokue Akihiko
Published: August 2003

Objective: The purpose of the present study was to clarify the clinicopathological significance of both thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) in renal cell carcinoma (RCC) based on a quantitative analysis of RCC patients. Methods: Levels of TP and DPD in RCC and/or uninvolved renal tissues from 65 RCC patients were measured by enzyme-linked immunosorbent assay. Results: The TP level and TP/DPD ratio were significantly higher in RCC than in adjacent uninvolved renal tissues (p < 0.0001). There was no significant difference in DPD levels between RCC and uninvolved renal tissues. The ratio of the highest to the lowest level was 623 in TP level, 28.9 in DPD level, and 985 in TP/DPD ratio. In the univariate analysis, patient’s age (p = 0.04), tumor stage (p < 0.0001), tumor size (p = 0.007), TP expression (p = 0.03), and DPD expression (p = 0.04) were significantly associated with increased risk of death. Multivariate analysis showed that patient’s age, tumor stage, and TP expression were independent prognostic factors. Conclusions: TP and DPD in RCC provide prognostic information although DPD was not an independent prognostic factor. The present finding of a wide range in these enzyme expressions in RCC suggests that a certain subpopulation with a high TP/DPD ratio has potential responsiveness to fluoropyrimidines, especially 5’-deoxy-5-fluorouridine and capecitabine.
Association between Serum Carcinoembryonic Antigen and Endothelial Cell Adhesion Molecules in Colorectal Cancer
Authors: Roselli Mario , Guadagni Fiorella , Martini Francesca , Spila Antonella , Mariotti Sabrina , D’Alessandro Roberta , Aloe Simona , Gazzaniga Pier Paolo , Basili Stefania , Cosimelli Maurizio , Ferroni Patrizia
Published: August 2003

Objectives: To analyse the behaviour of pre-surgical serum levels of soluble (s)E-selectin and vascular cell adhesion molecule (sVCAM) in patients with colorectal cancer, and to evaluate their possible correlation with carcinoembryonic antigen (CEA), pro-inflammatory cytokines and clinicopathological features with respect to their prognostic value in predicting metastatic disease. Methods: Pre-surgical serum levels of sE-selectin, sVCAM, interleukin-6 (IL-6), IL-1β, tumour necrosis factor-α (TNF-α) and CEA were measured in 194 patients with colorectal adenocarcinoma, 40 patients with benign colorectal diseases and 59 healthy subjects. Results: sE-selectin, sVCAM, TNF-α and IL-6 levels were significantly higher in patients with colorectal cancer compared to either healthy subjects or patients with benign disease. Positive rates of sE-selectin, sVCAM and TNF-α levels were significantly associated with Dukes’ stage D colorectal cancer, and all three variables were independently associated to the presence of distant metastases. Positive sE-selectin, sVCAM and TNF-α levels were significantly associated to CEA. TNF-α and CEA levels were independently related to the presence of positive levels of sE-selectin and/or sVCAM. Conclusions: Our findings suggest that the host inflammatory response to cancer cells, and/or their released products (i.e. CEA), might be responsible (via cytokine release) for the elevation in circulating adhesion molecules in patients with colorectal cancer.
Clinical Relevance of Sialyltransferases ST6GAL-I and ST3GAL-III in Gastric Cancer
Authors: Gretschel Stephan , Haensch Wolfgang , Schlag Peter M. , Kemmner Wolfgang
Published: August 2003

Aberrant glycosylation of membrane components due to specific alterations of glycosyltransferase activity is a common feature of carcinoma cells and is usually associated with invasion and metastasis. In a prospective study, the enzyme activity of the sialyltransferases ST6GAL-I and ST3GAL-III was studied in gastric cancer and normal mucosa in 55 patients by a radiometric assay. Cellular localization of sialyltransferase ST6GAL-I mRNA expression was studied by in situ hybridization. Sialyltransferase ST6GAL-I mRNA expression was mainly localized to epithelial cells. ST6GAL-I enzyme activity was enhanced within the tumor tissue. Significant correlations were found between the presence of signet ring cells and enhanced ST6GAL-I activity in the tumor tissue (p = 0.047) or in the mucosa (p = 0.024), and between signet ring cells and ST3GAL-III activity in the mucosa (p < 0.001). Multivariate Cox analysis demonstrated that only lymph node metastases (p = 0.044) had a significant influence on tumor-related survival. ST3GAL-III and ST6GAL-I activity showed no independent prognostic relevance in multivariate analysis, but high levels of ST3GAL-III and ST6GAL-I in the tumor tissue correlated with secondary local tumor recurrence (p = 0.005; p = 0.012). Interestingly, also the nonmalignant and uninvolved mucosa of tumor patients was altered on the molecular level and in some cases showed enhanced sialyltransferase levels indicative of the alteration of glycosylation very early during tumorigenesis.
Colorectal Cancer Relapse: Allelic Alterations Associated with Tumour Marker Overexpression
Authors: Vegh Irene , De-La-Cruz Javier , Navarro Susana , Morales Carlos , Colina Francisco , Abad Alfredo , De-La-Calle Angel , Enriquez-de-Salamanca Rafael , Moreno-González Enrique
Published: August 2003

The objective of the present study was to assess the prognostic value of allelic alterations in comparison with clinical prognostic factors (age and gender, clinical stage, lymph node involvement, tissue tumour marker expression) and clinical outcomes (disease relapse and overall survival time) in colorectal cancer patients. Polymerase chain reaction was performed on the DNA of 72 colorectal samples (from 36 colorectal cancer patients) using primers D17S513 and D17S514. Carbohydrate antigen 19-9 (CA 19-9) marker was determined in tumour sections by enzyme immunoassay. Tumours were considered to exhibit allelic alterations if the microsatellite region adjacent to the p53 locus in chromosome 17 either gained or lost repeated sequences. Allelic alterations were detected in 44% of tumour samples. Patients with more than 3 involved lymph nodes had more frequent allelic alterations (p < 0.002). The allelic alteration status was compared with tumour CA 19-9 expression, which showed statistically significantly higher values within the allelic alterations group (p < 0.005). Multivariate analyses confirmed that tumours with allelic alterations had a higher probability of disease relapse (odds ratio 7.3, p = 0.01). This is the first report showing an association between allelic alteration and overexpression of a tissue tumour marker protein and established risk factors. These results could be considered useful additional prognostic information for colorectal cancer.
Expression of Phosphatidylethanolamine N-Methyltransferase in Human Hepatocellular Carcinomas
Authors: Tessitore Luciana , Marengo Barbara , Vance Dennis E. , Papotti Mauro , Mussa Antonio , Daidone Maria Grazia , Costa Aurora
Published: August 2003

Objective: Hepatic phosphatidylethanolamine is converted into phosphatidylcholine by the enzyme phosphatidylethanolamine N-methyltransferase (PEMT) when the dietary choline supply is inadequate. Our previous reports implicated PEMT in the regulation of hepatocyte growth and transformation. In the present study, we analyzed PEMT activity, Pempt gene status and its mRNA expression in 29 human hepatocellular carcinomas (HCC). Methods: The status of the Pempt gene and PEMT2 mRNA expression were evaluated with Southern and Northern blotting, respectively, in HCC and the noninvolved liver. PEMT activity was assessed by biochemical assay. Cell proliferation markers were defined by immunohistochemical or histoautoradiographic methods. Results: PEMT activity was lower in HCC than in the noninvolved liver and it was negligible in 62% of the tumors. No deletions or mutations of the Pempt gene were found and PEMT2 mRNA expression was absent or reduced in HCC compared with peritumoral liver tissue. PEMT2 mRNA expression was inversely related to tumor proliferation and to histologic grade. Patients whose HCC did not express PEMT2 mRNA showed poorer outcomes for cancer-related survival than those with PEMT2-positive HCC. Conclusions: The present findings suggest that (1) clones lacking PEMT2 expression may have been selected during liver tumorigenesis and progression, and (2) PEMT2 expression seems to be associated with clinical progression.
Alteration of G2 Cell Cycle Regulators Occurs during Carcinogenesis of the Endometrium
Authors: Tsuda Hiroshi , Hashiguchi Yasunori , Inoue Takeshi , Yamamoto Kumio
Published: August 2003

Objectives: In this study, we examined the alteration of the G2 pathway in endometrial hyperplasia (EH) and endometrioid-type endometrial cancer (EC), and analyzed the relationship between the G2 pathway status and the p53 pathway status. Methods: A total of 103 cases (proliferative phase of the endometrium: 20, EH: 22, and endometrioid-type EC: 61 (I: 39, II: 5, III: 15, recurrence: 2)) were included in this study. We examined the ATM, chk2, CDC25C, cdc2, and cyclin B1 protein expression by immunohistochemistry. In 55 cases (EH: 15; EC: 40), we analyzed chk2 mutations by RT-PCR-SSCP. Results: There were no chk2 mutations in endometrial disease. Elevated or reduced expression rates of ATM, chk2, CDC25C, cdc2 and cyclin B1 were 4.5% (1/22), 0%, 0%, 0% and 4.5% (1/22) in EH and 3.3% (2/61), 4.9% (3/61), 13.1% (8/61), 9.8% (6/61) and 9.8% (6/61) in EC. Alteration of the G2 pathway was higher in EC (32.8%; 20/61) than in EH (9.1%; 2/22; p = 0.047). The G2 pathway was significantly higher in the altered p53 pathway group (48.4%; 15/31) than in the normal p53 pathway group (16.7%; 5/30) in EC (p = 0.0134). The altered p53 pathway tended to be related with the cdc2/cyclin B1 status (p = 0.0529). Conclusions: Alteration of the G2 pathway is thought to occur during carcinogenesis of the endometrium.
Interleukin-1α Enhances Integrin α6β1 Expression and Metastatic Capability of Human Pancreatic Cancer
Authors: Sawai Hirozumi , Funahashi Hitoshi , Yamamoto Minoru , Okada Yuji , Hayakawa Tetsushi , Tanaka Moritsugu , Takeyama Hiromitsu , Manabe Tadao
Published: August 2003

Objective: To investigate the mechanisms of metastasis formation in metastatic human pancreatic cancer, we examined the enhancement in integrin expression, and adherence to and invasiveness into extracellular matrix (ECM) proteins of human pancreatic cancer cells after exposure to interleukin (IL)-1α. Methods: Expression of IL-1 receptor type I (IL-1RI) and alterations in integrin subunits by IL-1α were examined by flow-cytometric analysis and by cellular enzyme-linked immunosorbent assay in four human pancreatic cancer cell lines (BxPC-3, PaCa-2, PANC-1, and SW1990), respectively. In addition, assays of cancer cell adhesion and invasion to ECM proteins were performed to investigate whether increased integrin expression affected the adhesive and invasive interaction between cancer cells and the putative integrin ECM ligands. Furthermore, immunohistochemistry was used to assess integrins and IL-1R1 expression in pancreatic tissues. Results: In metastatic cancer cells, expression of the α6 subunit was enhanced by IL-1α treatment. While metastatic cancer cells exhibited preferential adherence to and invasion into laminin, these properties were enhanced by IL-1α. The α6 subunit and IL-1RI were strongly expressed in pancreatic tissues from pancreatic cancer patients with liver metastasis. Conclusions: In pancreatic cancer, IL-1α enhanced α6β1-integrin expression, probably via increased IL-1RI levels. Our results indicated that α6β1-integrin and IL-1RI expression may play important roles in metastasis formation.
Adhesion and Migration of HRT-18 Colorectal Carcinoma Cells on Extracellular Matrix Components Typical for the Desmoplastic Stroma of Colorectal Adenocarcinomas
Authors: Hauptmann Steffen , Budianto Daniel , Denkert Carsten , Köbel Martin , Borsi Laura , Siri Annalisa
Published: August 2003

Colorectal carcinomas belong to the group of desmoplastic carcinomas which are characterized by an extensive connective tissue stromal component containing a variety of extracellular matrix (ECM) molecules. The biological importance of desmoplasia is still under discussion. Formerly, it has been regarded as walling off, thus being an obstacle for tumor cell invasion. In contrast, ECM may serve as a migratory substrate for tumor cells providing an advantage for invasion. Therefore, we have performed an in vitro investigation of the role of collagen types I, III, and V, fibronectin, and two variants of tenascin-C on adhesion and migration of the colorectal carcinoma cell line HRT-18. The data indicate that migration and adhesion strongly depend on both the composition and the concentration of ECM molecules. Even discrete changes in matrix composition can significantly modulate tumor cell migration, indicating that various degrees of invasiveness, e.g. at the tumor-host interface of colorectal carcinomas, can be attributed to environmental modifications.
Successful Treatment of Human Immunodeficiency Virus-Related Castleman’s Disease: A Case Report and Literature Review
Authors: Liberopoulos Evagelos , Tolis Christos , Bai Mary , Efremidis Stavros , Pavlidis Nikolaos , Elisaf Moses
Published: August 2003

Multicentric Castleman’s disease is increasingly recognized as an aggressive illness with a rapidly fatal outcome in human immunodeficiency virus (HIV)-infected patients. In the absence of optimal therapy, various therapeutic interventions have been tested with disappointing results; only five reports with a successful outcome have been described. Presented herein is a 66-year-old HIV-infected man with multicentric Castleman’s disease. Early administration of cyclophosphamide, doxorubicin, vincristine and prednisone resulted in prolonged clinical recovery. The relevant literature is also reviewed.
Protective Effect of Flavonoids against Aging- and Lipopolysaccharide-Induced Cognitive Impairment in Mice
Authors: Patil Chandrashekhar S. , Singh Vijay Pal , Satyanarayan P.S.V. , Jain Naveen K. , Singh Amarjit , Kulkarni Shrinivas K.
Published: August 2003

Flavonoids, naturally occurring polyphenolic compounds, are known to inhibit both lipopolysaccharide (LPS) stimulated tumor necrosis factor alpha and interleukin 6 release which modulate the proinflammatory molecules that have been reported in many progressive neurodegenerative disorders, including Alzheimer’s disease (AD), viral and bacterial meningitis, AIDS dementia complex, and stroke. The present experiments were performed to study the possible effects of exogenously administered flavonoids (apigenin-7-glucoside and quercetin) on the cognitive performance in aged and LPS-treated mice (an animal model for AD) using passive avoidance and elevated plus-maze tasks. Aged and LPS-treated mice showed poor retention of memory in step-through passive avoidance and in plus-maze tasks. Chronic administration of the flavonoids apigenin-7-glucoside (5–20 mg/kg i.p.) and quercetin (25–100 mg/kg i.p.) dose dependently reversed the age-induced and LPS-induced retention deficits in both test paradigms. However, flavonoids after chronic administration in young mice did not show any improvement of memory retention in both paradigms. Apigenin-7-glucoside showed more efficacy as compared with quercetin in both models that may be probably due to its greater efficacy to inhibit cyclooxygenase-2 and inducible nitric oxide synthase. Chronic treatment with flavonoids did not alter the locomotor activity in both young and aged mice; however, aged mice showed improvement of performance on Rota-Rod test. The results showed that chronic treatment with flavonoids reverses cognitive deficits in aged and LPS-intoxicated mice which suggests that modulation of cyclooxygenase-2 and inducible nitric synthase by flavonoids may be important in the prevention of memory deficits, one of the symptoms related to AD.
Differential Effects of Lidocaine and Tramadol on Modified Nerve Impulse by 4-Aminopyridine in Rats
Authors: Mert Tufan , Gunes Yasemin , Guven Mustafa , Gunay Ismail , Gocmen Cemil
Published: August 2003

We have used the sucrose gap method to measure the effects of drugs on the electrophysiological properties of rat sciatic nerves. The results showed that 4-aminopyridine produced a slight conduction block, prolonged the duration of action potential, enhanced the hyperpolarizing afterpotential, and elicited a hump that followed the action potential. In the presence of 4-aminopyridine, the impulse-blocking activity of lidocaine and tramadol was enhanced. Both lidocaine and tramadol effectively depressed the delayed depolarization generated by 4-aminopyridine. While tramadol decreased the activity-evoked hyperpolarizing afterpotentials, lidocaine completely removed them. These findings indicate that lidocaine may be more effective in blocking the Na+ channels than tramadol. Tramadol may be more effective on the delayed rectifier K+ channels than lidocaine.
Inducible Expression of Hypoxia-Inducible Factor 1α (HIF-1α) as a Tool for Studying HIF-1α-Dependent Gene Regulation during Normoxia in vitro
Authors: Meissner Udo , Allabauer Ida , Repp Reinald , Rascher Wolfgang , Dötsch Jörg
Published: August 2003

The hypoxia-inducible factor 1α (HIF-1α), a member of the PAS superfamily, is a global regulator of cellular and systemic O2 homeostasis as well as embryonic development. As the activity of HIF-1α is increased by a lowered oxygen tension in vivo and in vitro, we established a cell line producing high amounts of HIF-1α under normoxic conditions. As this overexpression was inducible by doxycycline, we can now provide a system to study HIF-1α-dependent gene regulation under normoxic as well as hypoxic conditions. We were able to show that the doxycycline-induced induction of the target gene HIF-1α – followed by the message of its target genes erythropoietin and vascular endothelial growth factor – is a dose- and time-dependent process. As the inducible overexpression of HIF-1α did not increase the rate of apoptosis, it provides a helpful new tool in drug discovery and tumor research to differentiate between hypoxia-dependent and hypoxia-independent pathways during HIF-1α-dependent gene regulation and HIF-1α-dependent effects on apoptosis.
Characterization of the Effect of Epilobium Extracts on Human Cell Proliferation
Authors: Vitalone Annabella , McColl Janice , Thome Dean , Costa Lucio G. , Tita Beatrice
Published: August 2003

We have previously shown that extracts of different Epilobium species, a phytotherapeutic agent used in folk medicine as a treatment for benign prostatic hyperplasia, inhibit proliferation of human prostate cells. The selectivity of this effect was evaluated in four different human cell lines (PZ-HPV-7, normal prostate cells; LNCaP, transformed prostate cells; HMEC, mammary cells, and 1321N1, astrocytoma cells). Different extracts of Epilobium species (E. rosmarinifolium, E. spicatum, and E. tetragonum) had similar growth-inhibitory effects in all cell lines tested, indicating a lack of specificity for prostate cells. Inhibition of DNA synthesis was mostly due to the nonpolar fraction of the extracts which is expected to contain flavonoids and sterols. Polar fractions were devoid of activity with the exception of that from E. rosmarinifolium. This species is the most potent in the antiproliferative effect and contains the highest concentration of oenothein B, a hydrolyzable ellagitannin. Oenothein B inhibited DNA synthesis in all four cell lines tested. Extracts of E. angustifolium (the Linné denomination of E. spicatum) and of E. spicatum from different sources were compared for their ability to inhibit DNA synthesis and for their oenothein B content. The E. angustifolium extract contained an amount of oenothein B 40-fold higher than the other extract of the same species and was ten times more potent in inhibiting DNA synthesis in a human prostate cell line. These results indicate that Epilobium extracts inhibit proliferation of prostate cells in a nonspecific manner. Oenothein B may play a role in this effect, but other active compounds are also present. The difference observed between extracts from the same species underscores the importance of determination and standardization of active ingredients in phytotherapeutic agents.
Effects of Some Antidepressant Drugs on Tryptaminergic Responses of the Rat Jejunum
Authors: Liu Haihuang , Coupar Ian M. , Irving Helen R.
Published: August 2003

The rat isolated jejunum was used as a functional model to screen some antidepressant drugs at the ‘atypical’ 5-HT7 receptor. Mianserin acted as a surmountable antagonist of 5-CT with a pA2 value of 8.22 ± 0.33. The apparent pKB of maprotiline against 5-CT was 7.67 ± 0.24 at 100 nmol/l, but higher concentrations suppressed the maxima. An apparent pKB could not be obtained for amitriptyline, because 10 nmol/l reduced the Emax of 5-CT without causing parallel displacement. Higher concentrations of 30 and 100 nmol/l caused further suppressions. Amoxapine, loxapine and desipramine (each at 100 nmol/l) caused similar effects, suppressing the Emax values to 5-HT by approximately 50%, while lower concentrations failed to cause parallel displacements. These results further extend the activity profiles of the drugs investigated.
Antinociceptive Effects of Methysergide in Various Pain Models
Authors: Chung Ki-Myung , Choi Seong-Soo , Han Ki-Jung , Han Eun-Jung , Lee Han-Kyu , Suh Hong-Won
Published: August 2003

Several studies have demonstrated that the nonselective opioid receptor antagonist naloxone produces a paradoxical antinociception in the formalin test. The opioid system is related to the serotonergic system for producing antinociception at the spinal level. Here we also asked whether systemic (i.p.) and intrathecal (i.t.) administrations of a nonselective serotonergic antagonist, methysergide, might produce paradoxical antinocicpetion similar to naloxone in the mouse formalin test. A diluted formalin solution was injected into the mouse plantar region of the hind paw and the duration of licking responses was measured at periods of 0–5 min (1st phase) and 20–40 min (2nd phase) after formalin injection. Methysergide administered i.p. and i.t. showed an attenuated licking duration only in the 2nd phase. The effect observed in the 2nd phase was reversed in the 5,7-dihydroxytriptamine, but not N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine pretreated group of mice, suggesting that descending serotonergic, but not noradrenergic, systems are involved in the methysergide antinociception. To further investigate the mechanism by which methysergide inhibited the nociceptive behaviors induced by formalin, the antinociceptive effect of methysergide was also tested in substance P (i.t.) and excitatory amino acids (i.t.), such as glutamate, N-methyl-D-aspartic acid, α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid, and kainic acid, which are major components in the formalin-induced nociceptive transmission in the spinal cord pain models. The duration of nociceptive behaviors shown in these models was significantly shortened by i.p. and i.t. administration of methysergide. These results suggest that methysergide also produces a paradoxical antinociception in various pain models including the formalin test, similar to the results of naloxone.
Pharmacokinetics of Isoflurane: Uptake in the Brain
Authors: Lu Chih-Cherng , Ho Shung-Tai , Wang Jhi-Joung , Wong Chih-Shung , Hu Oliver Yao-Pu , Chang Sun-Yran , Lin Chung-Yuan
Published: August 2003

We studied the effect of the inspired isoflurane concentration (CIiso) on the pharmacokinetics of isoflurane uptake in the brain by comparing the isoflurane concentration in internal jugular-bulb blood (Jiso) with that in arterial blood (Aiso), and analyzed this by gas chromatography. Sixteen patients (aged 43–76 years) undergoing colorectal surgery were enrolled, and anesthesia was maintained with a constant CIiso of either 1% (group 1, n = 8) or 2% (group 2, n = 8) during the 1st hour of isoflurane anesthesia. Under constant volume-controlled ventilation, we measured the CIiso and the end-tidal isoflurane concentration (CEiso) at the mouthpiece by infrared analysis. Our results demonstrate that it takes 40 min for the brain tissue concentration to equal Aiso for 1% CIiso, and 50 min for 2% CIiso. The Aiso (and/or Jiso) for 2% CIiso was approximately double when compared to that for 1% CIiso. Except during the initial wash-in period of the functional residual capacity in the first 3 min, the differences between CIiso and CEiso revealed that the body uptake of isoflurane for 2% CIiso was twice that for 1% CIiso. These results demonstrate that the pharmacokinetics of isoflurane uptake in the brain is time-dependent for Jiso to equal Aiso, and the midpoint between Aiso and Jiso (likely representing the isoflurane concentration in brain tissue) was dependent on CIiso.
4-Aminopyridine Transiently Increases Intraocular Pressure in Rabbits
Authors: Socci Robin R. , Chu Eugenia , Bayorh Mohamed A. , Chu Teh-Ching
Published: August 2003

The purpose of this study is to determine the mechanisms of action involved in the ocular hypertension induced by 4-aminopyridine (4-AP), a voltage-dependent potassium (K+) channel blocker, in rabbits. Topical application of 4-AP elevated intraocular pressure (IOP). This action caused increases in the aqueous flow rate as well as aqueous levels of protein and norepinephrine. In isolated iris-ciliary body preparations, 4-AP (0.01, 0.1, 1 mmol/l) caused dose-related increases in field-stimulated norepinephrine release by 43, 222 and 243%, respectively. Taken together, the IOP-elevating effect evoked by 4-AP was associated with enhancement of aqueous norepinephrine levels and norepinephrine release from sympathetic nerves of the iris-ciliary body. These results demonstrate that the 4-AP-sensitive K+ channels in sympathetic nerves and the ciliary epithelium are the potential sites of action of the 4-AP-induced ocular hypertension.
Characterization of Sialidase from an Influenza A (H3N2) Virus Strain: Kinetic Parameters and Substrate Specificity
Authors: França de Barros Jr. José , Sales Alviano Daniela , da Silva Maria Helena , Dutra Wigg Marcia , Sales Alviano Celuta , Schauer Roland , dos Santos Silva Couceiro José Nelson
Published: August 2003

Neuraminidase (NA) of influenza A (H3N2) viruses was characterized after purification by gel filtration and proteolytic treatment, using the X-31 variant strain that is a reassortment between the influenza A/Victoria/3/75 (responsible for the 1975 pandemic) and the influenza A/PR/8/34 virus samples, as a model. In the purification process, NA heads, that is the spike responsible for the virus sialidase activity, were purified by filtration through a Bio-Gel polyacrylamide column. The enzyme activity was determined by periodic acid/thiobarbituric acid assay and high-performance thin-layer chromatography. The sialidase showed preference for the α-2,3-linkage over the α-2,6-linkage of sialyllactoses (Km of 1.8 and 5.2 × 10–4M, respectively) at pH 5.2. The enzyme acted on natural and synthetic substrates at different hydrolysis rates, as well as on human erythrocytes (A group, Rh+) and yeast (Candida albicans) cells. The active NA produced by gel filtration was characterized by different parameters of its sialidase activity, also showing to be a suitable tool for the identification of natural sialocompounds and for the screening of antisialidase drugs to treat influenza virus infections.
Molecular Diversity of Partial-Length Genomic Segment 2 of Human Picobirnavirus
Authors: Martínez Laura C. , Giordano Miguel O. , Isa Maria B. , Alvarado Luis Fernando , Paván Jorge V. , Rinaldi Diego , Nates Silvia V.
Published: August 2003

Objectives: This study was carried out in order to evaluate the efficacy of the recently developed picobirnavirus (PBV) sets of primers and to establish the phylogenetic relationships of Argentine strains with PBV strains isolated in China and the USA. Methods: Thirteen fecal specimens tested as positive for PBV by polyacrylamide gel electrophoresis were analyzed by reverse transcription-polymerase chain reaction assays using primers target to the genomic segments 2 of PBV strains isolated in China and the USA. The amplicons were sequenced and analyzed. Results: Primers derived from the China strain produced amplicons in only 4 of the 13 specimens (30.76%). No sample was revealed as positive with the primers derived from the US strain. DNA sequencing of polymerase chain reaction products differed in nucleic acid and amino acid sequences by 13.9–42.28% and 18.1–51.1%, respectively. Despite this strain diversity, three domains of conserved nucleotide sequences as well as the amino acid motif D-S-D typical of RNA-dependent RNA polymerase gene of double-strand RNA viruses were identified. Comparatively, these conserved regions were also identified in homologous PBV strains from the USA and China. Phylogenetic analysis showed no time or geographic clustering. Conclusions: These findings provide evidence that PBV may represent an emerging heterogeneous group of viruses.
TT Virus Infection Does Not Affect the Clinical Profiles of Patients with Hepatitis B and C in Yanbian City, China
Authors: Moriyama Mitsuhiko , Longren Wu , Zi-Yi Zhao , Oshiro Shuh , Matsumura Hiroshi , Aoki Hiroshi , Shimizu Toshihiro , Nakai Kazuhiko , Yamagami Hiroaki , Kaneko Miki , Shioda Atsuo , Ichijima Sagiri , Iwaguchi Kayo , Iwasaki Hiroko , Tanaka Naohide , Arakawa Yasuyuki
Published: August 2003

China is an area of high endemicity for viral hepatitis, and the molecular epidemiological investigation of TT virus (TTV) infection is of interest. In the present study, we investigated the epidemiology, clinical significance and molecular characteristics of TTV infection in patients with chronic hepatitis B and C in Yanbian City, China. Serum samples obtained from 74 patients with hepatitis B and hepatitis C who visited Yanbian Hospital, located in northeast China, were analyzed in this study. The study group included 22 cases of chronic hepatitis B (B-CH), 17 cases of liver cirrhosis B (B-LC), 7 cases of hepatocellular carcinoma (B-HCC), 16 cases of chronic hepatitis C (C-CH), 11 cases of liver cirrhosis C (C-LC) and 1 case of hepatocellular carcinoma (C-HCC). Detection of TTV DNA was performed as described by Nishizawa et al. The second-round PCR products from 7 subjects were sequenced, followed by investigation of nucleotide homology and phylogenetic analysis. TTV DNA was present in 18.2, 5.9, 28.6, 6.3, 9.1 and 0% of the patients with B-CH, B-LC, B-HCC, C-CH, C-LC and C-HCC, respectively. The highest prevalence of TTV infection was seen in the groups aged 40–50 and over 60 years. There was no significant correlation between the presence of TTV DNA and the clinical parameters in patients with hepatitis B and C. The various isolates showed 97.9–100% with isolates reported previously from Japan and 98.4–100% with isolates reported previously from China. Nucleotide sequence analysis revealed that the Yanbian isolates could be classified in the same group as the Japan and China isolates. We concluded that chronic coinfection with TTV did not affect the serological features of chronic hepatitis B and C in China, as found in Tokyo, Japan.
Low HBeAg Serum Levels Correlate with the Presence of the Double A1762T/G1764A Core Promoter Mutation and a Positive Response to Interferon in Patients with Chronic Hepatitis B Virus Infection
Authors: Marrone A. , Zampino R. , Luongo G. , Utili R. , Karayiannis P. , Ruggiero G.
Published: August 2003

Objective: To investigate the correlation of serum hepatitis B e antigen (HBeAg) levels with the presence of core promoter (CP) mutations, hepatitis B virus (HBV) viremia and the response to interferon (IFN) in patients with chronic hepatitis B. Methods: Fourteen HBeAg-positive patients received α-2a IFN. Diluted serum samples of responders were tested for HBeAg positivity at dilutions of 1:40, 1:160 and 1:640 at the following time points: T0 (before starting IFN), T1 [at peak alanine aminotransferase (ALT) preceding HBeAg seroconversion], T2 (at ALT normalisation) and T3 (end of treatment). Nonresponder samples were similarly tested at times T0 and T3. The HBV CP and precore regions were sequenced at the same time points as for HBeAg testing. Results: Six of 14 patients (43%) responded to IFN treatment and had lower HBeAg levels than nonresponders at T0 (p = 0.003). Five of 6 responders (83%) and none of the nonresponders had the A1762T/G1764A CP mutations (0/8, p < 0.003). At T0, HBeAg was negative at the 1:640 dilution in 5 of the 6 responders, who also had lower HBV DNA levels than nonresponders (p = 0.003). During IFN treatment, HBeAg levels decreased and HBV DNA became negative at T1 in responders. Conclusions: Low serum HBeAg and HBV DNA levels correlate with the presence of CP mutations and response to IFN treatment and can be considered as predictive markers of response to IFN.
Improved Detection of Dengue-1 Virus from IgM-Positive Serum Samples Using C6/36 Cell Cultures in Association with RT-PCR
Authors: Oliveira De Paula Sérgio , Malta Lima Danielle , Clotteau Marilidia , Pires Neto Roberto da Justa , Lopes da Fonseca Benedito Antônio
Published: August 2003

Dengue is the most important arboviral disease in the world, and its diagnosis is primarily made by serology. Virus isolation has been successful mainly in clinical samples obtained during the acute phase of illness, and is carried out through inoculation of clinical samples into C6/36 cell monolayers followed by the detection of infection by indirect immunofluorescence assay (IFA). We compared the efficiency of RT-PCR and IFA in the detection of dengue-1 virus after inoculation of C6/36 cells with samples obtained in the convalescent period of dengue infection. Out of 75 IgM-positive samples inoculated into C6/36 cells, 2 were positive by IFA while 17 were positive by RT-PCR. The 2 IFA-positive samples were collected during the acute phase of the illness; 17 positive samples were found by RT-PCR, including the 2 detected by IFA. For both methods, we also investigated the time necessary for viral detection using a fixed dose of 1 × 104 viruses/ml. RT-PCR and IFA detected the dengue virus 1 and 4 days after virus inoculation, respectively. The results obtained here indicate that RT-PCR is the most sensitive method in the detection of dengue viruses using C6/36 cells for viral isolation.
Laboratory Diagnosis of Norovirus: Which Method Is the Best?
Authors: Rabenau H.F. , Stürmer M. , Buxbaum S. , Walczok A. , Preiser W. , Doerr H.W.
Published: August 2003

Noroviruses (NV) are transmitted by fecally contaminated food, vomit, and person-to-person contact. They are one of the main causes of non-bacterial acute gastroenteritis in nursing, old people and children’s homes. NV outbreaks are characterized by a short incubation period (12–48 h), nausea, vomiting and diarrhea, and high secondary attack rates. The illness is generally mild and self-limiting. The aim of diagnostic procedures in viral gastroenteritis is to avoid nosocomial infections on the one hand and unnecessary antibiotic treatment on the other. Diagnostic procedures for NV are based on the detection of virus in stool samples by (immune) transmission electron microscopy (TEM), antigen ELISA, or polymerase chain reaction (PCR). In our study, a total of 244 stool samples obtained from 227 patients between March and May 2002 were tested by TEM, antigen ELISA and in-house PCR. Our data showed that PCR has the highest sensitivity (94.1%), followed by TEM (58.3%), and ELISA (31.3%), while specificity was highest for TEM (98.0%), followed by ELISA (94.9%), and PCR (92.4%). All three methods tested (TEM, ELISA and PCR) are useful for epidemiological investigations in gastroenteritis outbreaks; however, to maximize diagnostic validity for individual cases, at least two of the methods should be combined.
The Tat Protein of the Caprine Arthritis Encephalitis Virus Interacts with the Notch2 EGF-Like Repeats and the Epithelin/Granulin Precursor
Authors: Shoham Nitza , Cohen Limor , Gazit Arnona , Yaniv Abraham
Published: August 2003

Using the yeast two-hybrid system, we screened a human placenta cDNA library and identified two proteins that interacted with the Tat protein of the caprine arthritis encephalitis virus (CAEV): the EGF-like repeats 1–6 of the extracellular domain of the human Notch2 receptor and the epithelin/granulin growth factor precursor. This interaction was also confirmed in mammalian cells. Using in vitro mutagenesis assays, we showed that each one of the three cysteine residues located within the cysteine-rich domain of the CAEV Tat protein is essential for the binding of Tat to both the Notch2 and the epithelin/granulin protein. It is thus suggested that the cysteine-rich domain of Tat plays a role in the interaction between the Tat and either Notch2 or the epithelin/granulin domains, both of which exhibit EGF-like-repeat-imposed spatial conformation. It is assumed that such interactions might modulate the physiological functions of Notch2 and epithelin/granulin, thereby affecting various pathologies associated with CAEV.
Characteristics of Apoptotic Cell Death Induced by Coxsackievirus B in Permissive Vero Cells
Authors: Ahn Jeonghyun , Joo Chul-Hyun , Seo Ilsun , Kim DongHou , Hong Hea Nam , Kim Yoo Kyum , Lee Heuiran
Published: August 2003

Coxsackievirus B (CVB) causes a wide spectrum of human diseases which are closely associated with direct destruction of infected cells. We investigated the morphological and biochemical characteristics of CPEs in permissive Vero cells caused by different CVB serotypes. Regardless of serotype, the infected cells experienced similar degrees of CPEs within 24 h postinfection (p.i.). Using both Hoechst 33342 staining and transmission electron microscopy, we consistently observed morphological properties of apoptosis, heavily condensed nuclei and subsequent chromatin condensation into the periphery of the nuclei within 12 h p.i. Moreover, we noticed typical oligonucleosomal DNA fragmentation, while productive CVB multiplication was accomplished within 6 h p.i. prior to an apoptotic signal. Caspase inhibitor significantly prohibited nuclear changes due to apoptosis with no influence on virus production and cell death, demonstrating that all the CVBs induced more than one type of pathological effect, including apoptotic alteration in permissive Vero cells.
A Genotype IV Hepatitis E Virus Strain That May Be Indigenous to Changchun, China
Authors: Liu Ziling , Chi Baorong , Takahashi Kazuaki , Mishiro Shunji
Published: August 2003

We sequenced the RNA genome of hepatitis E virus (HEV) recovered from an acute hepatitis patient living in Changchun, China. The HEV-CCC220 isolate thus obtained was 7,193 nt in length and contained ORF1 (5,121 nt, 1,706 aa), ORF2 (2,025 nt, 674 aa), and ORF3 (345 nt, 114 aa) plus uncompleted 5′ and 3′ untranslated regions. When compared with full-genome isolates so far reported, the CCC220 isolate segregated to genotype IV although the nucleotide similarity to the previous genotype IV isolates was only 83.9–85.6%. Comparison of a partial sequence (300 nt) within the ORF2 between the genotype IV isolates from China, Taiwan and Japan also indicated the phylogenetic uniqueness of the CCC220 isolate: even the highest homology was less than 90%. These findings suggest that the CCC220 isolate might represent HEV strains that are indigenous to Changchun, China.
Evolution of Forebrain and Spatial Cognition in Vertebrates: Conservation across Diversity
Authors: Salas Cosme , Broglio Cristina , Rodríguez Fernando
Published: August 2003

Historically the dominant trend in comparative brain and behavior research has emphasized the differences in cognition and its neural basis among species. In fact, the vertebrate forebrain shows a remarkable range of diversity and specialized adaptations. Probably the major morphological variation is that observed in the telencephalon of the actinopterygian fish, which undergoes a process of eversion during embryonic development, relative to the telencephalon of non-actinopterygians (for instance, amniotes), which develops by a process of evagination. These different developmental processes produce notable variation, mainly two solid telencephalic hemispheres separated by a unique ventricle in the actinopterygian radiation that contrasts with the hemispheres with internal ventricles in other groups. However, an increasing amount of evidence reveals that the forebrain of vertebrates, whether everted or evaginated, presents a common pattern of basic organization that supports highly conserved cognitive functions. We analyze here recent data indicating a close functional similarity between spatial cognition mechanisms in different groups of vertebrates, mammals, birds, reptiles, and teleost fish, and we show in addition that they rely on homologous neural mechanisms. Thus, recent functional and behavioral comparative evidence is added to the developmental and neuroanatomical data suggesting that the evolution of cognitive capabilities and their neural basis in vertebrates could have been more conservative than previously realized.
Comparative Psychology of Surprising Nonreward
Authors: Papini Mauricio R.
Published: August 2003

The surprising or unexpected omission of an appetitive reinforcer has at least two effects: An allocentric effect according to which the organism updates knowledge about the environment, and an egocentric effect that allows the organism to learn about its own emotional reaction to the change. This egocentric effect (traditionally called frustration) is correlated to activation of the hypothalamic-pituitary-adrenal axis, can be modulated by treatment with anxiolytics, and is expressed in terms of behavioral changes that have an emotional component (e.g., agonistic behavior). It is hypothesized that all vertebrates share the mechanisms underlying the allocentric effect, but only mammals possess the mechanisms underlying the egocentric effect. It is further argued that frustrative mechanisms evolved in early mammals from those underlying fear conditioning.