bluuebunny/crossref_metadata_2025 huggingface dataset:
Reduced Expression of GNA11 and Silencing of MCT1 in Human Breast Cancers
Authors: Asada Kiyoshi , Miyamoto Kazuaki , Fukutomi Takashi , Tsuda Hitoshi , Yagi Yukiko , Wakazono Kuniko , Oishi Shoko , Fukui Hiroshi , Sugimura Takashi , Ushijima Toshikazu
Published: May 2003

Alteration in the methylation status of a gene is often associated with its altered expression. Based on a genome scanning technique for differences in CpG methylations, methylation-sensitive representational difference analysis, DNA fragments hypermethylated in a human breast cancer were isolated. A DNA fragment was isolated from intron 1 of guanine-nucleotide-binding protein α-11 (GNA11). mRNA expression of GNA11 was shown to be decreased in 10 of 16 breast cancers by RT-PCR analysis, and the immunoreactivity of the GNA11 product, Gα11 subunit of heterotrimeric G-protein, was observed to be reduced in 14 of the 16 cancers by immunohistochemistry. Methylation of a CpG island (CGI) in the 5′ region of GNA11 or that of intron 1 did not show a clear correlation with its decreased expression. Another DNA fragment was isolated from a CGI in the 5′ upstream region of monocarboxylate transporter 1 (MCT1), and was methylated in 4 of 20 breast cancers. The CGI was also methylated in a human breast cancer cell line, MDA-MB-231, and quantitative RT-PCR showed that its expression was almost lost in the cell line. By treatment of the cells with a demethylating agent, 5-aza-2′-deoxycytidine, the methylation was removed and the expression was restored. GNA11 is involved in signalling of gonadotropin-releasing hormone receptor, which negatively regulates cell growth. MCT1 is involved in cellular transportation of butyrate, which induces cellular differentiation. Downregulation of these two genes was suggested to be involved in human breast cancers.
Prognostic Role of Apoptotic, Bcl-2, c-erbB-2 and p53 Tumor Markers in Salivary Gland Malignancies
Authors: Nagler Rafael M. , Kerner Hedviga , Ben-Eliezer Shoshana , Minkov Ira , Ben-Itzhak Ofer
Published: May 2003

Objective: The clinical characteristics and survival probability rate of 36 patients with salivary gland malignancies and 10 patients with benign salivary tumors were summarized in relation to the immunohistological analysis of the tumor, apoptotic-related markers and apoptosis rate. The expression of the markers examined – Bcl-2, c-erbB-2, p53 – was detected in paraffin sections of the tumors by the streptavidin-biotin peroxidase method following heat-induced antigen retrieval, and the apoptosis rate was determined by the TUNEL method. Results: The overall 5-year survival probability was 61% for patients with malignant tumors and 100% for those with benign tumors. The survival probability of patients over 60 at diagnosis was significantly lower than that of younger patients. Patients whose malignant tumors were larger than 2 cm at diagnosis had worse survival than those with smaller tumors. The survival probability of patients whose malignant tumors were located in the submandibular glands was significantly lower than that of patients whose malignancies were located in the parotid and minor salivary glands. The survival probability of patients who demonstrated positive staining for c-erbB- 2 or TUNEL was lower than for those with negative staining. Gender, the existence of concomitant non-salivary malignancies and ethnic origin had no significant impact on survival. Conclusions: Our results demonstrated significant positive staining in the salivary tumorigenic tissue but not in the surrounding non-tumorigenic tissue examined for TUNEL, c-erbB-2, Bcl-2 and p53, pointing to a biological role for all four markers in the tumorigenic process which is yet to be elucidated. Significant reduction in survival was related to the specific location of the tumor in the submandibular gland, its size and older age of patient. Survival was also found to be significantly reduced when positive staining was demonstrated in the tumor tissue for TUNEL or c-erbB-2, more so for concomitant positive staining of both markers. Clinically, the most important result of the current study is that the survival rate of the patients examined with salivary tumors larger than 2 cm, with positive staining for both TUNEL and c-erbB-2, was 0 (p = 0.0001)!
Sensitivity of Non-Small-Cell Lung Cancer Cell Lines Established from Patients Treated with Prolonged Infusions of Paclitaxel
Authors: Fujishita Takashi , Loda Massimo , Turner Ross E. , Gentler Marinshine , Kashii Tatsuhiko , Breathnach Oscar S. , Johnson Bruce E.
Published: May 2003

Objective: Regimens with prolonged infusions of taxanes have been developed for patients with cancer to overcome drug resistance. Our objective of the present study was to examine the impact of prolonged exposure on the cytotoxicity of taxanes against non-small-cell lung carcinoma (NSCLC) cell lines and the clinical response and outcome of the patients. Methods: Five cell lines (NCI-H2882, -H2887, -H2973, -H3122, -H3255) were derived from previously untreated patients with NSCLC who participated in clinical trials of continuous 96-hour infusions of paclitaxel followed by bolus cisplatin. Two additional cell lines (NCI-H838, -H1299) with previously published data were used as controls. Drug sensitivities were assessed by the MTS (Promega) assay. Multidrug resistance (MDR) phenotypes were assessed by quantitative real-time PCR and HER-2/neu by both immunohistochemistry and ELISA. Results: The median of mean IC50 values of docetaxel at the exposure durations of 3, 24, 72 and 120 h were 0.52, 0.06, 0.03 and 0.06 µM, respectively. The median of mean IC50 values of paclitaxel at the exposure duration of 3, 24, 72 and 120 h were 0.48, 0.13, 0.03 and 0.02 µM, respectively. In all cell lines studied, there was a less than 4-fold difference in the IC50 values between docetaxel and paclitaxel at 3-, 72-, and 120-hour exposure times. The single cell line with moderate MDR1 expression (NCI-H2887) was the only cell line established from a patient with progressive disease when treated with paclitaxel. Conclusions: This study demonstrates prolonged exposure to both docetaxel and paclitaxel inhibits the growth of NSCLC cell lines in similar fashion.
Identification of a Novel Homeobox-Containing Gene, LAGY, Which Is Downregulated in Lung Cancer
Authors: Chen Yuan , Petersen Simone , Pacyna-Gengelbach Manuela , Pietas Agnieszka , Petersen Iver
Published: May 2003

We have isolated a novel gene, lung cancer-associated gene Y (LAGY), by suppression subtractive hybridization. The nucleotide sequence of LAGY predicts a small protein of 73 amino acids containing a putative homeobox domain with a molecular mass of 8.1 kD. Multiple-tissue Northern blot analysis revealed that LAGY is present in human placenta, lung, brain, heart and skeletal muscle. Gene mapping locates LAGY on chromosome 4q11–13.1. The expression of LAGY mRNA was widely lost in 18 lung tumor cell lines comprising all major histological types, as shown by Northern blot analysis and semiquantitative reverse transcription-polymerase chain reaction. In an investigation of 72 primary lung tumors, this gene was significantly downregulated in tumors compared to 9 normal lung tissue samples. There was a significant reduction of LAGY expression in squamous cell carcinoma (SCC; n = 27) with increasing grade and stage. No expression was detectable in two high-grade SCCs or two small cell and large cell lung carcinomas (n = 4 for each). In adenocarcinoma (n = 37), expression was reduced; however, this did not reach statistical significance. Since homeodomain-containing genes are known to transcriptionally regulate key cellular processes and are associated with carcinogenesis, we suggest that LAGY might be linked to lung cancer development and progression.
Peritoneal Carcinoma in a Male Patient
Authors: Jermann Monika , Vogt Peter , Pestalozzi Bernhard C.
Published: May 2003

Peritoneal carcinoma is a rare primary tumor, described in the literature almost exclusively in women. This report describes our clinicopathological findings in a 51-year-old male patient with peritoneal carcinoma and ascites. Pathologic studies included routine histology, immunohistochemistry and electron microscopy on biopsy and autopsy tumor tissue. After chemotherapy, the patient achieved a complete remission twice, lasting for 14 months and 8 months, respectively, and died after 3 years. His clinical course was similar to that of female patients with peritoneal carcinoma or advanced ovarian cancer. Our case confirms the existence of primary peritoneal carcinoma in males. In addition, it shows that this entity responds to the same chemotherapy as used for ovarian cancer and primary peritoneal carcinoma in females.
Obsessive-Compulsive Disorder in Childhood and Adolescence
Authors: Presta S. , Marazziti D. , Dell’Osso L. , Pfanner C. , Pfanner P. , Marcheschi M. , Masi G. , Muratori F. , Mucci M. , Millepiedi S. , Cassano G.B.
Published: May 2003

Obsessive-compulsive disorder (OCD) is a common psychiatric condition during childhood and adolescence, which continues to be underestimated and undertreated. For these reasons, it constitutes a primary cause of major disabilities in those ages and, sometimes, of permanent impairments later on. In these last few years, childhood and adolescence OCD has attracted an increasing focus which has promoted a deeper awareness of this illness, a better recognition with earlier interventions, as well as the set-up of more tailored and specific strategies, including psychotropic drugs. The aim of this paper is to present a critical review of paediatric OCD, with a special attention towards the most compelling reports available up to now and towards the most interesting areas for future research.
A Study of the Phenomenology of Panic Attacks in Patients from India
Authors: Neerakal I. , Srinivasan K.
Published: May 2003

We studied the symptom structure of panic attacks in 94 patients attending an outpatient psychiatric clinic. Autonomic symptoms of anxiety were reported by the majority of the panic patients. As compared with studies from the West, certain cognitive symptoms, such as fear of loss of control and depersonalization, and vestibular symptoms were endorsed by fewer panic patients. The majority of our panic patients tended to seek help initially from physicians and medical specialists. The majority of the patients also did not experience some DSM-IV symptoms as part of their panic attacks. Comorbid psychiatric conditions influenced the symptom structure of panic attacks. A higher number of agoraphobics as compared with patients with uncomplicated panic disorders had the symptoms of fear of loss of control and fainting. Our findings support the notion that the experience of panic attacks is variable and differs across cultures.
Amplification and Attribution Styles in Somatoform and Depressive Disorders – A Study from Bangalore, India
Authors: Duddu V. , Chaturvedi S.K. , Isaac M.K.
Published: May 2003

Objective: The present investigation aimed to study attribution styles and somatosensory amplification among patients suffering from somatoform and depressive disorders. Methods: Two groups of 30 patients with diagnoses of somatoform disorder and depressive disorder, respectively (ICD-10 DCR), and one group of 30 normal controls were recruited. The study patients were assessed using the symptom interpretation questionnaire, somatosensory amplification scale, and scales for assessing alexithymia and illness attitudes. Results: The somatoform and depressive disorder patients had greater recent symptom experience than the normal group. The somatoform disorder group had higher somatic attribution scores, the depressive disorder sample had higher psychological attribution scores, and the normal group had higher normalizing attribution scores than the two other groups. Somatoform disorder patients had higher mean amplification scores than depressed patients, who in turn had higher scores than normals. Correlation analyses showed somatic attribution and certain illness attitudes to be closely associated in all three groups. Recent symptom experience was associated with amplification in the somatoform disorder group alone. Recent symptom experience, a diagnosis of somatoform disorder and lower normalizing attribution scores predicted amplification. Discussion: These findings indicate that somatoform and depressive disorder patients and normals differ from each other in their attribution styles. There is a clustering of attributes among somatoform disorder patients that include greater symptom experience, which is somatically attributed, and is associated with excessive illness worry, concern and preoccupation with bodily symptoms, and a fear of having or developing a disease. On the other hand, depressed patients and normal subjects who do have a somatic attribution style (though, as a group, they have lower somatic attribution scores than the somatoform disorder group), also harbor hypochondriacal beliefs and related attitudes.
Changes in Duodenal Mast Cells in Response to Dehydroleucodine
Authors: Penissi Alicia , Rudolph Isolde , Fogal Teresa , Piezzi Ramón
Published: May 2003

Introduction: Dehydroleucodine (DhL), a sesquiterpene lactone isolated from Artemisia douglasiana Besser, prevents gastroduodenal damage elicited by necrosis-inducing agents such as absolute ethanol. Changes in the number of mast cells or evidence of activation of the cells for mediator release have been observed in a wide spectrum of disease processes involving the gastrointestinal tract. In the present study we examined the effects of DhL on duodenal mast cell population and their histamine content, with the goal of throwing more light on the mechanism of action of the drug. Materials and Methods: Male Rockland mice (n = 30) were divided into two groups and administered orally with 0.4% carboxymethylcellulose (CMC; control group) or DhL in 0.4% CMC, 40 mg/kg body weight (DhL group). The animals were killed 60 min after dosing and their duodena were removed. Mucosal and submucosal mast cells were studied by light and electron microscopy, and the duodenal histamine content was measured by high-performance liquid chromatography. Results: DhL increased the number of mast cells in the submucosal layer. This was related to an increase in the tissue histamine levels (from 324 ± 19.14 to 1,284 ± 20 pg/mg tissue, in controls and DhL-treated, respectively). The mast cells in the submucosa from the control group showed a cytoplasm containing a predominant population of homogenously dense granules, and the DhL-treated group exhibited swollen granules showing different degrees of particulation. The mucosal mast cell population showed no modifications in response to the cytoprotective agent. Conclusions: DhL induces (1) a selective increase in the number of mast cells in the submucosal layer and (2) changes in the distribution and appearance of their secretory granules. These findings, probably associated with the higher histamine levels after DhL treatment, could be involved in the cytoprotective action of the lactone, previously reported by us.
Sleep Deprivation Inhibits Expression of NADPH-d and NOS while Activating Microglia and Astroglia in the Rat Hippocampus
Authors: Hsu Jee-Ching , Lee Ying-Shiung , Chang Chen-Nen , Chuang Huo-Li , Ling Eng-Ang , Lan Chyn-Tair
Published: May 2003

This study investigated the expression of nitric oxide (NO)-synthesizing enzymes and the glial reaction in the rat hippocampal formation following sleep deprivation for 5 days. Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) reactivity was markedly reduced in the hippocampal CA1, CA2 and CA3 sectors as well as in the dentate gyrus, suggesting a suppression of NO production in these areas. Microglial cells were hypertrophic and showed an up-regulation of complement type 3 receptors as determined by antibody OX-42. However, expression of major histocompatibility complex class I and II antigens, and antigen of monocyte/macrophage lineage marked by OX-18, OX-6 and ED1, respectively, was undetected. Astrocytes also displayed hypertrophied processes with enhanced glial fibrillary acidic protein (GFAP) immunoreactivity. Western blots of hippocampal tissues corroborated the above-mentioned morphological findings in that expression of NO-synthase (NOS) was decreased while that of OX-42 and GFAP was increased in the sleep-deprived rats. Since NO is thought to be involved in memory consolidation processes in the hippocampus during sleep, the inhibition of NADPH-d and NOS reactivities may account for the memory decline after long-term sleep deprivation. The concomitant reactions in microglia and astrocytes suggest the involvement of these cells in the deleterious effect of prolonged sleep deprivation.
Curative Surgery for Local Pelvic Recurrence of Rectal Cancer
Authors: Saito Norio , Koda Keiji , Takiguchi Nobuhiro , Oda Kenji , Ono Masato , Sugito Masanori , Kawashima Kiyotaka , Ito Masaaki
Published: May 2003

Background/Aims: Local pelvic recurrence of rectal cancer after radical resection has been associated with morbidity and cancer-related death. This study retrospectively evaluated outcome following curative resection for rectal cancer recurring after surgery on the basis of prognosis, type of procedure and perioperative morbidity. Methods: A total of 85 consecutive patients with local pelvic recurrence of rectal cancer were evaluated. Of these, 43 underwent microscopic curative surgery for local recurrence. Among the 43 patients, 23 underwent surgery alone and 17 received preoperative radiotherapy (40 Gy) (XRT group) in addition to the surgery. Of the 43 patients, 26 were asymptomatic. Results: Curative resection was higher in the recurrences that were associated with implantation, incomplete surgical margin clearance, and intrapelvic lymph node metastasis than in other types of recurrence. With regard to surgical procedure, abdominoperineal resection (APR), with or without sacral resection, was standard following previous sphincter-preserving surgery, while total pelvic exenteration (TPE), with or without sacral resection, was common following previous APR. Local excision was not considered appropriate surgery. There was a high incidence of perioperative morbidity (64%) in patients receiving TPE. Re-recurrence was observed in 18 patients (50%) after curative surgery. After a follow-up of 2 years or more, the local re-recurrence rate was 28%. The overall 5-year survival rate for patients receiving curative resection was 39%, for patients in the XRT group, 51%, and for patients in the surgery-alone group, 24% (p = 0.07). The survival rate in 26 asymptomatic patients was higher than in 17 patients with symptoms, with 5-year survival rates of 62 and 23% (p < 0.05), respectively. The cumulative local control in the preoperative radiotherapy plus en bloc surgery group (XRT group) was significantly better than in the surgery-alone group (p < 0.01), and survival in the XRT group tended to be better than in surgery alone. Conclusions: These results suggest that careful patient selection according to the pattern of recurrence, area of invasion and presence of symptoms is important for successful curative surgery. Aggressive surgery with adjuvant therapy may lead to an improved salvage rate.
Influence of the Route of Reconstruction on Morbidity, Mortality and Local Recurrence after Esophagectomy for Cancer
Authors: Wong Alfred C.C. , Law Simon , Wong John
Published: May 2003

Background: A choice of retrosternal or orthotopic route for reconstruction exists after three-phase esophagectomy with cervical anastomosis. Whether the route of reconstruction affects postoperative morbidity, mortality and recurrence patterns remains controversial. Study Design: Patients with cancer of the thoracic esophagus who underwent three-phase esophagectomy between 1990 and 1999 were studied. Postoperative outcome, recurrence patterns and survival were analyzed from a prospectively collected database. Results: Seventy-five patients underwent three-phase esophagectomy. There were 46 patients in the retrosternal group and 29 in the orthotopic group. The mean age of the retrosternal group was younger than the orthotopic group, 60 and 66 years, respectively (p = 0.004). The retrosternal group also had more advanced disease; 24% of patients had curative resection compared with 59% in the orthotopic group (p = 0.003). There was no significant difference in postoperative complications except that the retrosternal group had more blood loss, median 800 ml compared with 700 ml (p = 0.04). Hospital mortality was 13% in the retrosternal group and 3.4% in the orthotopic group (p = 0.24). Multivariate analysis showed that age (odds ratio 1.16, p = 0.035) and pulmonary risk (odds ratio 10, p = 0.01) were predictive of hospital mortality, but not the route of reconstruction. No patient in the retrosternal group developed recurrence in the gastric conduit compared to 4 of 28 patients (14%) in the orthotopic group (p = 0.03). Two of these patients were symptomatic with bleeding from the intragastric recurrence. Survival was worse in the retrosternal group, 5-year survival was 29.8 vs. 8.2% (p < 0.01), reflecting the more advanced disease and higher prevalence of palliative resections. Conclusions: Cardiopulmonary complications and hospital mortality were not significantly different in the two groups. Recurrent tumor infiltration of the gastric conduit occurred in 14% of patients when the orthotopic route was used.
Implications of Removing a Normal Appendix
Authors: Bijnen C.L. , van den Broek W.T. , Bijnen A.B. , de Ruiter P. , Gouma D.J.
Published: May 2003

Background: The diagnosis of acute appendicitis remains difficult and therefore 15–30% of the removed appendices appear to be normal. The aim of this study was to investigate morbidity, mortality and costs of removing a normal appendix in patients with suspected appendicitis. Patients and Methods: Retrospective study of patients who underwent a negative appendectomy for suspected appendicitis in the period 1991–1999 with a median follow-up of 4.4 years. Patients who underwent an elective appendectomy or appendectomy for other reasons were excluded. Results: In 285 patients (70% women, 30% men) a normal appendix was removed. In 192 (67%) patients a muscle-splitting incision was performed, in 6 (2%) a median laparotomy, and in 51 (18%) the normal appendix was removed by laparoscopy. In 36 patients (13%) a diagnostic laparoscopy was converted to a muscle-splitting incision. Complications occurred in 16 (6%) patients, in 5 (2%) a re-operation was needed. The mean hospital stay was 4.4 (SE 2.8) days, in case of complications 7.4 (SE 4.2) days. The mean extra hospital costs of a negative appendectomy were EUR 2,712. Conclusion: The removal of a normal appendix has considerable complications and costs. In an attempt to prevent these costs, extra diagnostic tools should be considered. Expensive diagnostic tools such as diagnostic laparoscopy should be used selectively in order not to further increase costs.
Use of Photoplethysmography to Determine Gastrointestinal Perfusion Pressure: An Experimental Canine Model
Authors: Garcia-Granero E. , Garcia S.A. , Alos R. , Calvete J. , Flor-Lorente B. , Willatt J. , Lledo S.
Published: May 2003

Objectives: To develop an experimental model to assess the parietal perfusion pressure (PPP) of the digestive tract using photoplethysmography. Materials and Methods: Twenty-two mongrel dogs were used. Progressive external compression was applied to the intestinal wall and the PPP was assessed with photoplethysmography. The study group was divided into two groups. In group 1 PPP was measured at the levels of the stomach, duodenum, jejunum and transverse colon. In group 2 PPP was measured after temporary occlusion of the truncal and marginal circulation of the jejunum to provide further variables. Results: The PPP decreased significantly for each successive distal section. Correlation coefficients and indices for PPP and mean arterial pressures were statistically significant (p < 0.005). Truncal occlusion provoked a drop in PPP whereas marginal occlusion scarcely modified the basal results. Conclusions: Photoplethysmography, through measurements of the residual arterial wave amplitude, is a valid method of determining quantitatively the PPP of the digestive tract and could be useful in a clinical environment.
Effects of the Free Radical Scavenger Dimethyl Sulphoxide on Experimental Normothermic Ischaemia of the Liver
Authors: Chiappa Antonio , Makuuchi Masatoshi , Zbar Andrew P. , Biella Francesca , Bellomi Massimo , Biffi Roberto , Bertani Emilio , Vezzoni Aldo , Crosta Cristiano , Andreoni Bruno
Published: May 2003

Background/Aims: This study assessed the effects of intermittent or continuous hepatic ischaemia and reperfusion with or without dimethyl sulphoxide (DMSO) pre-treatment in a rat ischaemic model. Methods: One hundred and eighty rats were divided into three groups undergoing hepatic ischaemia of a total duration of 60, 90 and 120 min. Each group of rats was subdivided to receive either a continuous Pringle manoeuvre or intermittent liver pedicle clamping of 30 or 15 min. Ten minutes before ischaemia induction, 10 rats from each group were pre-treated with DMSO (500 mg/kg, b.w.) intravenously. Results: With continuous hepatic pedicle clamping, survival rates inversely correlated with the duration of ischaemia, with greater survival in the intermittently clamped groups (p < 0.05). DMSO pre-treatment did not affect survival but resulted in a significant reduction in liver enzyme (aspartate aminotransferase, alanine aminotransferase) release on the first postoperative day following total ischaemic times of 90 min or greater (p < 0.05). After 120 min of total ischaemia, DMSO pre-treatment resulted in higher preservation of adenosine 5′-triphosphate liver content (p < 0.05). Conclusion: DMSO may be used to prolong tolerance to inflow occlusion and to limit the adverse effects of ischaemia and reperfusion cycles in an experimental hepatic ischaemia model.
Children with Growth Hormone Deficiency and Chiari I Malformation: A Morphometric Analysis of the Posterior Cranial Fossa
Authors: Tubbs R. Shane , Wellons III John C. , Smyth Matthew D. , Bartolucci Alfred A. , Blount Jeffrey P. , Oakes W. Jerry , Grabb Paul A.
Published: May 2003

Objective: The posterior fossa (PF) has been found to be small in various forms of Chiari malformation. Explanations involving a connection between growth hormone deficiency (GHD) and Chiari I malformation (CIM) have been proposed. However, to date, no quantitative analysis of the PF of patients with CIM and GHD has been performed. Our study was performed to determine the geometry of the PF in children with GHD and CIM. Methods: Morphometric analysis of the PF was performed in 10 children with GHD and CIM (group 1), 20 children with GHD and no CIM (group 2) and 50 controls. Results: PF volumes for group 1 ranged from 128 to 259 ± 33 ml, and for group 2, they ranged from 115 to 186.2 ± 25.4 ml. Lengths of the foramen magnum for groups 1 and 2 had means of 36 and 38 mm, respectively. The mean basiocciput length and tentorial angle for groups 1 and 2 were 20 and 19 mm and 89 and 87.5°, respectively. Conclusions: We have determined that children with GHD with or without CIM have no significant difference in their PF volume compared to controls. However, our data demonstrate significant underdevelopment of portions of the bony PF in both patients with GHD alone and in patients with GHD and CIM. Tentorial angles were elevated in noncontrol groups. We propose that this association is not due to an increased rate of ‘midline’ defects seen in GHD but rather a structurally distorted PF that is not capacious enough to house the entire developing rhombencephalon. These data will hopefully aid in the further understanding of the pathophysiology of CIM.
Alterations of Thrombogenesis, Endothelial Damage and Oxidative Stress with Reperfusion during Femoral Artery Bypass Surgery for Peripheral Vascular Disease
Authors: Makin Andrew J. , Chung Natali A.Y. , Silverman Stanley H. , Lip Gregory Y.H.
Published: May 2003

Peripheral vascular disease (PVD) is a significant cause of cardiovascular morbidity. We hypothesised that there would be significant alterations of thrombogenesis, platelet activation and endothelial damage, which could be associated with abnormal oxidative stress during femoral artery bypass surgery for PVD, where the femoral artery is cross-clamped (causing acute ischaemia) and reperfused (following revascularisation). To test this hypothesis, we measured sequential changes in von Willebrand factor (vWF, and index of endothelial damage/dysfunction), tissue factor (TF, an index of thrombogenesis) and soluble P-selectin (sP-sel, an index of platelet activation) as well as lipid hydroperoxides (LPO, an index of oxidative stress) in 28 consecutive patients undergoing elective peripheral artery bypass surgery. Mean baseline vWF and sP-sel levels in PVD patients (before clamping) were significantly higher compared with age- and sex-matched controls (unpaired t test, both p < 0.05), but there were no significant differences in TF and LPO levels. There was a correlation between TF and vWF (Spearman’s, r = 0.374, p = 0.05), as well as between sP-sel and vWF at the start of surgery (r = 0.467, p = 0.012). The patients undergoing peripheral artery bypass surgery had a mean femoral artery clamp time of 28 min (standard deviation 14 min; range 11–65 min). There were no significant overall changes in sP-sel, vWF, TF and LPO with femoral artery cross-clamping and reperfusion (repeated measures ANOVA, p = NS). In conclusion, we found that during ischaemia-reperfusion during peripheral arterial bypass surgery, thrombogenesis (as measured by plasma TF) and oxidative damage (as measured by LPO) within the affected leg does not increase in the immediate perioperative period. Further studies are required to assess the mechanism(s) of ischaemia-reperfusion injury in PVD, and the contributory role(s) of the endothelium and platelets.
Bound Thrombin from Crushed Clots Is Composed of α-Thrombin and the N-Terminal Regions of α- and γ-Chains of Fibrinogen
Authors: Kinjoh Kiyohiko , Nakamura Mariko , Gang Zeng , Sunagawa Masanori , Eguchi Yukinori , Kosugi Tadayoshi
Published: May 2003

We aimed at clarifying the structural characteristics of the bound thrombin that is liberated by mechanical breakdown of fibrin clots. Fibrin clots were prepared with bovine thrombin and rabbit fibrinogen, and were crushed mechanically with a glass rod. The supernatant of the crushed clots was subjected to immunoaffinity chromatography to isolate the bound thrombin. Western blotting analysis revealed that the bound thrombin could be reacted with both antithrombin and antifibrinogen under unreduced conditions. SDS-PAGE under reduced conditions revealed that there were three bands, two of which were found to be the N-terminal fragments of the α- and γ-chains of fibrinogen. The bound thrombin could be dissociated into three distinct fibrin fragments and bovine α-thrombin when denatured by 8 M urea. Thus, the bound thrombin liberated from crushed clots is a stable complex between bovine α-thrombin and fibrin fragments of the N-terminal regions of rabbit α- and γ-chains.
New Method for the Extraction of DNA from White Blood Cells for the Detection of Common Genetic Variants Associated with Thrombophilia
Authors: Angelini Antonio , Di Febbo Concetta , Rullo Agnese , Di Ilio Carmine , Cuccurullo Franco , Porreca Ettore
Published: May 2003

In the last few years, FV Leiden, prothrombin 20210 and thermolabile methylene tetrahydrofolate reductase (MTHFR) 677 variants have been identified as possible risk factors associated with thrombophilia, and many PCR-based methods have been described for detecting these variations. However, the genomic DNA extraction is still a rate-limiting and time-consuming step in the PCR process. In an attempt to accelerate this procedure and make it suitable for routine laboratory, we report a single preparative technique for DNA extraction from peripheral blood samples using an anion-binding resin (GeneFizz). This method enables white blood cell lysis, DNA extraction and PCR amplification directly in the thermocycling tube on the thermocycler. The use of this new DNA extraction system coupled to a multiplex PCR allows rapid genetic screening of large cohorts of patients for thrombophilic risk factors.
Plugged-Percutaneous Liver Biopsy in Patients with Impaired Coagulation and Ascites
Authors: Kamphuisen Pieter W. , Wiersma Tjeerd G. , Mulder Chris J.J. , de Vries Richard A.
Published: May 2003

In liver cirrhosis coagulation is impaired due to decreased synthesis of vitamin K-dependent and vitamin K-independent coagulation factors. In such patients routine liver biopsy is contraindicated due to the increased risk of bleeding. Treatment with recombinant factor VIIa or fresh frozen plasma reduces the complication rate of liver biopsy, but both have disadvantages. In this observational study, we evaluated the safety and efficacy of plugged-percutaneous liver biopsy in 36 patients with ascites (n = 9), impaired coagulation (n = 22), or both (n = 5) due to severe chronic liver disease. Among patients with clotting disorders, mean prothrombin time was 16.3 s (range 11.4–20.3) and the mean platelet count was 53 × 109/l (range 19–153). Plugged-percutaneous liver biopsy was in none of the cases associated with bleeding complications (95% confidence interval 0–0.097). All biopsies were adequate for histological interpretation and therefore diagnostically successful. In our experience, plugged-percutaneous liver biopsy seems a safe and reliable method in patients with chronic liver disease associated with impaired coagulation and/or ascites needing histological evaluation.
Impact of Pollen on Human Health: More Than Allergen Carriers?
Authors: Traidl-Hoffmann Claudia , Kasche Anna , Menzel Annette , Jakob Thilo , Thiel Martina , Ring Johannes , Behrendt Heidrun
Published: May 2003

The transfer of pollen from floral anther to recipient stigma is the critical reproductive event among higher plants – this is the botanical view of pollen. Proteins and glycoproteins from pollen can function as allergens, environmental molecules interacting with the human immune system to elicit an allergic response in susceptible individuals – this is how allergists and immunologists see pollen grains. Between 10 and 25% of the population now have symptoms of hay fever or allergic asthma and the incidence has more than doubled in the past three decades while the reason(s) for this increment are only hypothetical, but there is a multitude of them. Despite our natural focus on this impact of pollen on human health, pollen have to be considered in a larger context. First of all, to evaluate the bioavailability of allergens from pollen, we have to understand their function and their influence factors. Furthermore, pollen grains are not only releasing proteins eliciting specific immune responses, but they also liberate bioactive lipid mediators and this much more rapidly. And last but not least, recent observations indicate, that pollen do not only induce allergy and thus have a much broader impact on human health. This review is an attempt to favour this holistic view of pollen and their impact on human health.
Inflammatory Role of Two Venom Components of Yellow Jackets (Vespula vulgaris): A Mast Cell Degranulating Peptide Mastoparan and Phospholipase A1
Authors: King Te Piao , Jim Sui Yee , Wittkowski Knut M.
Published: May 2003

Background: Venom sac extract of yellow jackets Vespula vulgaris was toxic in mice when injected intraperitoneally but not toxic when injected subcutaneously. Necropsy showed the toxicity to be an inflammatory response. Methods: Venom peptide and protein fractions were tested to identify the inflammatory components. The active components were tested to establish whether they might function as adjuvant for venom protein-specific antibody response. Results: Venom toxicity required the synergistic action of two venom components, a mast cell degranulating peptide mastoparan and phospholipase A1. Both components stimulated prostaglandin E2 release from murine peritoneal cells and macrophages. Mastoparan showed a weak activity to enhance IgE and IgG1 responses to a yellow jacket venom protein Ves v 5 in BALB/c mice. It was not possible to assess the adjuvant activity of phospholipase A1 because of its suppression of Ves v 5-specific response. Melittin, a mast cell degranulating peptide from bee venom, was inactive as an adjuvant for Ves v 5-specific response. Conclusion: Yellow jacket venom contains two inflammatory components, mastoparan and phospholipase A1. Our findings suggest that mastoparan can function as a weak adjuvant for TH2 cell-associated antibody response.
Analysis of Natural History of Japanese Cedar Pollinosis
Authors: Okawa Toru , Konno Akiyoshi , Yamakoshi Takayuki , Numata Tsutomu , Terada Nobuhisa , Shima Masayuki
Published: May 2003

Background: The marked increase in the incidence of Japanese cedar (Cryptomeria japonica; JC) pollinosis is a social problem in Japan. Elucidation of its natural history is, therefore, essential. Methods: Cross-sectional and vertical-sectional studies were performed regarding the effects of aging on sensitization to Japanese cedar pollen (JCP) and development of JC pollinosis by measuring serum IgE titers to JCP and by oral examination of residents of the Maruyama Town, Chiba, Japan from 1995 to 2001. We also studied the incidence of its spontaneous remission and the background factors. Results: In a vertical-sectional study, the serum IgE titer to JCP was strongly influenced by the amounts of pollen scattered. An increase in age by 6 years did not reduce the IgE titer to JCP in subjects in their 40s. However, in subjects aged 60 or more, annual differences in the JCP count did not affect serum IgE titer to JCP, which remained low even after a season with a high pollen count. In subjects with JC pollinosis aged over 40 showing a CAP RAST score of more than 2 to JCP in 1995, spontaneous remission of JC pollinosis was observed in 16.1% over a period of 6 years. Factors affecting spontaneous remission include sex, age at the time, serum IgE titer to JCP and age at first onset of JC pollinosis. Conclusions: The CAP RAST score was strongly associated with spontaneous remission in the multivariable model.
The Roles of Cysteinyl Leukotrienes in Eosinophilic Inflammation of Asthmatic Airways
Authors: Nagata Makoto , Saito Keiko
Published: May 2003

Cysteinyl leukotrienes (CysLTs), including LTC4, LTD4 and LTE4, are potent biological mediators generated from arachidonic acid and are produced by inflammatory cells, including eosinophils. Classically, CysLTs have been recognized as powerful spasmogens for bronchial smooth muscle and thus have been implicated in the pathogenesis of asthma. There is increasing evidence that CysLTs also contribute to accumulation of eosinophils within asthmatic airways; CysLTs have been reported to be chemotactic for eosinophils both in vitro and in vivo. CysLTs are also able to enhance the survival of eosinophils. Moreover, LTD4 augments eosinophil adhesion via β2 integrins to intercellular cell adhesion molecule (ICAM)-1, which is constitutively expressed on airway epithelium. Interaction with ICAM-1 enhances the effector functions of eosinophils including the release of specific granule proteins, initiation of the respiratory burst, and generation of additional CysLTs. Thus, CysLTs can augment both accumulation and activation of eosinophils, and may thereby contribute to the phenotypic changes observed in airway eosinophils. We also discuss the effect of CysLT receptor antagonists on airway inflammation in asthma.
Analysis of Gene Expression in Peripheral Blood Eosinophils from Patients with Atopic Dermatitis by Differential Display
Authors: Hashida Ryoichi , Ogawa Kaoru , Miyagawa Masami , Sugita Yuji , Takahashi Eiki , Nagasu Takeshi , Katsunuma Toshio , Akasawa Akira , Tsujimoto Gozoh , Matsumoto Kenji , Saito Hirohisa
Published: May 2003

To identify the genes related to atopic dermatitis (AD), we compared gene expression in eosinophils from AD patients and healthy volunteers. RNA was prepared from peripheral blood eosinophils. Gene expression was monitored by fluorescent differential display (DD) and real-time RT-PCR. Eighteen new sequences, including expressed sequence tags (ESTs), were expressed at higher levels in eosinophils from AD patients than in those from healthy volunteers. The functions of most of these genes are unknown. We found no correlation between the expression of a particular gene and clinical markers such as the number of eosinophils and the amount of IgE. Multivariate analysis of the gene expression data in each sample showed a very high coefficient of correlation among the copy numbers of each gene. The genes under investigation were also expressed in cultured blood eosinophils after IL-4, IL-5 and IFN-γ stimulation. We were able to predict the function of some of the sequences by scanning for homologies within either the human or mouse genome databases. The mouse counterpart of one of these genes, intersectin 2, was expressed dramatically, as measured by ear edema, in 1-fluoro-2,4-dinitrobenzene-induced mouse contact dermatitis and in NC/Nga mouse dermatitis.
VNTR Polymorphism of Tyrosine Hydroxylase Gene and Schizophrenia in the Korean Population
Authors: Pae Chi-Un , Kim Jung-Jin , Serretti Alessandro , Lee Chang-Uk , Lee Soo-Jung , Lee Chul , Paik In-Ho
Published: May 2003

This study aimed to test the association of the tyrosine hydroxylase (TH) gene with schizophrenia in the Korean population. 334 patients with schizophrenia and 391 healthy volunteers were included. Intron 1 VNTR polymorphism of TH gene was genotyped using polymerase chain reaction techniques. The genotype and allele distribution between patients and controls were not significantly different. However, the positive score of the Positive and Negative Syndrome Scale (PANSS) was higher in the group expressing the TH10 allele (t = 2.245, p = 0.02), although no significant differences were present in the distribution of age of onset, degree of improvement using Brief Psychiatric Rating Scale (BPRS) change, total PANSS score and negative score. This finding suggests that the TH10 allele may be a liability factor for positive schizophrenia in the Korean population.
Diffusion Tensor Magnetic Resonance Imaging of Disruption of Regional White Matter in Schizophrenia
Authors: Minami T. , Nobuhara K. , Okugawa G. , Takase K. , Yoshida T. , Sawada S. , Ha-Kawa S. , Ikeda K. , Kinoshita T.
Published: May 2003

Diffusion tensor imaging provides a new approach for quantifying anisotropic diffusion of white matter in vivo. We used this technique to investigate subtle disruption of regional white matter in schizophrenia. Twelve patients with schizophrenia were compared with 11 healthy controls. Psychotic symptoms were assessed using the Positive and Negative Syndrome Scale. A significant fractional anisotropy (FA) reduction was found in all white matter regions bilaterally in schizophrenic patients. Higher FA of left frontal white matter correlated significantly with higher dosage of antipsychotic medication. These findings support the view that the pathological process is a distortion of the central nervous system myelination affecting the whole white matter. Our findings also show the effects of antipsychotics on the white matter in the left frontal region in schizophrenia.
Serum Cholesterol in Treatment-Resistant Depression
Authors: Papakostas George I. , Petersen Timothy , Sonawalla Shamsah B. , Merens Wendelien , Iosifescu Dan V. , Alpert Jonathan E. , Fava Maurizio , Nierenberg Andrew A.
Published: May 2003

Objective: Patients with major depressive disorder (MDD) may have significant differences in cholesterol levels compared with healthy controls. A previous study by our group reported that depressed patients with elevated cholesterol levels (≧200 mg/dl) were significantly more likely to be nonresponders to fluoxetine treatment than depressed patients with nonelevated cholesterol levels. However, very little is known regarding cholesterol in patients with treatment-resistant depression (TRD). The purpose of this study was to compare cholesterol levels at baseline between depressed patients with and without TRD and to test whether cholesterol levels at baseline can predict clinical response in patients with TRD treated with open-label nortriptyline (NT). Methods: Ninety-two patients with TRD entered a 6-week open trial of NT. Baseline cholesterol levels were randomly collected for 59 of these patients. Controlling for age and gender, we compared baseline cholesterol and triglyceride levels for 35 patients with TRD who did not respond to NT with 205 non-TRD patients who responded to an 8-week open trial of fluoxetine. Furthermore, with the use of logistic regression, we tested whether baseline cholesterol levels predicted clinical response to NT in the patients with TRD. Results: Patients with TRD had higher triglyceride levels at baseline compared with depressed patients without TRD. Cholesterol defined as a dichotomous variable being elevated if equal to or greater than 200 mg/dl, predicted poor response to a 6-week open trial of NT in patients with TRD. Conclusions: The results of this study confirm the relationship between hypercholesterolemia and poor outcome in the treatment of MDD for patients with TRD.
Catecholamine and HPA Axis Dysfunction in Depression: Relationship with Suicidal Behavior
Authors: Pitchot William , Reggers Jean , Pinto Emmanuel , Hansenne Michel , Ansseau Marc
Published: May 2003

A large body of evidence suggests a potential role for catecholaminergic function as a possible biological factor in the control of suicidal behavior. Recently, we have used a neuroendocrine strategy to study dopaminergic and noradrenergic activities in depressed suicide attempters. However, some problems are associated with the use of growth hormone (GH) response to catecholaminergic challenge, because GH release could be decreased by a direct effect of corticosteroids at the pituitary level. Therefore, the purpose of the present study was to assess GH response to both apomorphine, a dopaminergic agonist, and clonidine, an α2-adrenergic agonist, according to the dexamethasone suppression test (DST) status in a sample of 20 major depressed inpatients with a history of suicide attempt compared with nonattempters. Our results tended to show that hypercortisolemia as assessed by post-DST cortisol values did not inhibit GH response to apomorphine or clonidine, suggesting that hypothalamo-pituitary-adrenal axis overactivity does not explain the impaired GH response to apomorphine in major depressed patients with a history of suicide attempt.
Fine Mapping of the Schizophrenia Susceptibility Locus on Chromosome 1q22
Authors: Brzustowicz Linda M. , Hayter Jared E. , Hodgkinson Kathleen A. , Chow Eva W.C. , Bassett Anne S.
Published: May 2003

Schizophrenia is a serious neuropsychiatric illness estimated to affect approximately 1% of the general population. As part of a genome scan for schizophrenia susceptibility loci, we have previously reported a maximum heterogeneity four-point lod score of 6.50 on chromosome 1q21-22 in a group of 22 medium-sized Canadian families, selected for study because multiple relatives were clinically diagnosed with schizophrenia or schizoaffective disorder. We have now conducted fine mapping of this locus in the same set of individuals using 15 genetic markers spanning an ∼15-cM interval. Parametric linkage analysis with GENEHUNTER v2.1 and VITESSE v2.0 produced a maximum multipoint heterogeneity lod score of 6.50, with a Zmax-1 support interval of <3 cM, corresponding to ∼1 Mb. Physical mapping and sequence analysis from this region confirmed the presence of an ∼81-kb tandem duplication, containing low-affinity IgG receptor genes and heat shock protein genes. The sequences of the two copies of this duplication are ∼97% identical, which has led to the collapse of the two copies into one in the June 2002 NCBI Build 30 of the Human Genome. This duplication may be involved in genomic instability, leading to gene deletion, and so presents an intriguing candidate locus for schizophrenia susceptibility.