bluuebunny/crossref_metadata_2025 huggingface dataset:
Joint cytokine quantification in two rodent arthritis models: kinetics of expression, correlation of mRNA and protein levels and response to prednisolone treatment
Authors: RIOJA I , BUSH K A , BUCKTON J B , DICKSON M C , LIFE P F
Published: June 2004

Biomarker quantification in disease tissues from animal models of rheumatoid arthritis (RA) can help to provide insights into the mechanisms of action of novel therapeutic agents. In this study we validated the kinetics of IL-1β, TNF-α and IL-6 mRNA and protein expression levels in joints from DBA/1OlaHsd murine collagen-induced arthritis (CIA) and Lewis rat Streptococcal cell wall (SCW)-induced arthritis by real-time polymerase chain reaction (PCR) TaqMan® and Enzyme-linked immunosorbent assay (ELISA). Prednisolone was used as a reference to investigate any correlation between clinical response and cytokine levels at selected time-points. To our knowledge this is the first report showing a close pattern of expression between mRNA and protein for IL-1β and IL-6, but not for TNF-α, in these two models of RA. The kinetics of expression for these biomarkers suggested that the optimal sampling time-points to study the effect of compounds on both inflammation and cytokine levels were day 4 postonset in CIA and day 3 after i.v challenge in SCW-induced arthritis. Prednisolone reduced joint swelling through a mechanism associated with a reduction in IL-1β and IL-6 protein and mRNA expression levels. At the investigated time points, protein levels for TNF-α in arthritic joints were lower than the lower limit of detection of the ELISA, whereas mRNA levels for this cytokine were reliably detected. These observations suggest that RT-PCR TaqMan® is a sensitive technique that can be successfully applied to the quantification of mRNA levels in rodent joints from experimental arthritis models providing insights into mechanisms of action of novel anti-inflammatory drugs.
Increased Fas-mediated apoptosis in polymorphonuclear cells from HIV-infected patients
Authors: SALMEN S , TERÁN G , BORGES L , GONCALVES L , ALBARRÁN B , URDANETA H , MONTES H , BERRUETA L
Published: June 2004

Neutrophils represent an important line of innate host defence against invading microorganisms and their functional detriment during HIV infection, including accelerated spontaneous cell death, has been shown to contribute to AIDS development. Neutrophils are susceptible to apoptosis via Fas and an interaction between Fas and FasL was suggested originally as a mechanism to explain constitutive neutrophil apoptosis. We have explored some intracellular pathways leading to PMN apoptosis from 28 HIV-infected patients and 24 healthy volunteers. As previously reported, accelerated spontaneous apoptosis was observed in HIV+ patients, but this did not correlate with viral load. Furthermore, an increase in the level of spontaneous apoptosis was detected in neutrophils from HIV-infected patients following inhibition of ERK, suggesting an impairment of this kinase pathway during the early stages of infection which may contribute to PMN dysfunction. An elevated susceptibility to undergo apoptosis was observed following cross-linking of Fas, which correlated both with viral load and co-expression of Fas/FasL surface molecules. Different mechanisms for spontaneous and Fas-induced apoptosis are proposed which together contribute to the neutropenia and secondary infections observed during the progression to AIDS.
Decreased interferon-gamma response in respiratory syncytial virus compared to other respiratory viral infections in infants
Authors: ABERLE J H , ABERLE S W , REBHANDL W , PRACHER E , KUNDI M , POPOW-KRAUPP T
Published: June 2004

An inappropriate interferon-gamma response has been implicated in the pathogenesis of severe respiratory syncytial virus (RSV) lower respiratory tract illness (LRTI). To assess whether this is unique for RSV primary LRTI compared to a first non-RSV LRTI, intracellular interferon-gamma was determined by flow cytometry in peripheral blood mononuclear cells from 32 infants with a primary RSV infection, 28 with a first non-RSV LRTI due to adenoviral, parainfluenzaviral and rhinoviral infection and 13 healthy infants. Interferon-γ responses were increased significantly during adenoviral, parainfluenzaviral and the majority of the rhinoviral infections, but remained low during RSV and severe rhinoviral infection. Low interferon-γ responses were associated with a more severe clinical course of LRTI. This indicates that depending on the nature of the viral pathogen, respiratory virus infections in infants differ significantly with regard to the quantity of the interferon-γ production and that this may contribute to the clinical course of the disease.
Rotavirus-specific T-cell responses in young prospectively followed-up children
Authors: MÄKELÄ M , MARTTILA J , SIMELL O , ILONEN J
Published: June 2004

Rotavirus is a major cause of gastroenteritis in young children. Antibodies seem to protect against rotavirus infection but cell-mediated immune responses are probably also important for protection. We evaluated the development of T-cell responses to rotavirus in follow-up samples from 20 healthy children with an increased genetic risk for type 1 diabetes. Blood samples from 16 healthy adults were also available for the study. T-cell proliferation was analysed at 3–6 month intervals from the age of 3 months to the age of 4–5 years using the Wa strain of human rotavirus and the NCDV strain of bovine rotavirus as antigens. IgG and IgA antibodies to rotavirus were studied from simultaneously drawn plasma samples with EIA method using NCDV as an antigen. A total of 24 infections were revealed by antibody analysis. Sixteen children showed diagnostic increases in both IgG and IgA antibodies to rotavirus, while 5 children showed increases in IgA antibodies only and 3 in IgG only. Antibody rises were accompanied by T-cell responses to rotavirus (SI > 3) in 9 of the 24 cases. T-cell responses to purified or lysed human rotavirus were stronger after a rise in rotavirus antibodies than the responses before infection (P = 0·017 and 0·027, respectively). There was a correlation between T-cell responses to purified and lysed human rotavirus and NCDV. Strong T-cell responses to rotavirus were transient and the ability to respond usually disappeared in one year, but in all adults T-cell responses to rotavirus were strong implicating that several infections are needed to develop consistent, strong T-cell responsiveness.
Differential regulation of mast cell cytokines by both dexamethasone and the p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580
Authors: KORANTENG R D , SWINDLE E J , DAVIS B J , DEARMAN R J , KIMBER I , FLANAGAN B F , COLEMAN J W
Published: June 2004

Activated mast cells generate multiple cytokines but it is not known if these can be differentially regulated by pharmacological agents. We report here that the glucocorticoid dexamethasone (DEX) preferentially inhibited Ag-induced expression of IL-4 and IL-6 mRNA relative to TNF-α mRNA in RBL-2H3 cells. Likewise, the drug more readily inhibited release of IL-4 than TNF-α protein. SB203580, an inhibitor of p38 mitogen-activated protein kinase (MAPK), enhanced Ag-induced TNF-α mRNA expression without affecting IL-4 or IL-6 mRNA. At the protein level, SB203580 exerted little effect on TNF-α release but inhibited IL-4 release; notably, the ratio of TNF-α : IL-4 increased markedly with the concentration of SB203580, confirming the differential regulation of these cytokines. PD98059, an inhibitor of MAPK kinase (MEK), a component of the p44/42 MAPK pathway, partially inhibited Ag-induced expression of mRNA for all three cytokines while cyclosporin A inhibited Ag-induced IL-4 and IL-6 mRNA more readily than TNF-α mRNA. Ag activation of the cells led to phosphorylation of p38 and p44/42 MAPK but this was not influenced by DEX. In conclusion, mast cell cytokines can be differentially regulated pre- and post-translationally by DEX and SB203580 but there does not appear to be a direct mechanistic link between the actions of these two drugs.
Polyreactive antigen-binding B (PAB+) cells are widely distributed and the PAB+ population consists of both B-1+ and B-1– phenotypes
Authors: ZHOU Z-H , NOTKINS A L
Published: June 2004

B cells that make polyreactive antibodies (PAB+ cells) express polyreactive Ig receptors on their surface and can bind a variety of different antigens. The present study shows that PAB+ cells are widely distributed, are present in varying numbers in different lymphoid organs and that their phenotype varies depending on the organs from which they are isolated. Up to 10 times more cells in PAB+ enriched populations bind antigens as compared to PAB– populations. Comparison of PAB+ with B-1+ cells showed that a high percentage of PAB+ cells are B-1+, but that many PAB+ cells do not express B-1 cell surface markers and, in fact, are B-1–. It is concluded that the B cell population consists of PAB+/B-1+, PAB+/B-1–, PAB–/B-1+, and PAB–/B-1– cells. The presence of PAB+ cells in the thymus points to the possibility that PAB+ cells may carry endogenous host antigens from peripheral tissues to the thymus where they may contribute to immunological tolerance.
Iron down-regulates macrophage anti-tumour activity by blocking nitric oxide production
Authors: HARHAJI L , VUCKOVIC O , MILJKOVIC D , STOSIC-GRUJICIC S , TRAJKOVIC V
Published: June 2004

Although the inhibitory effect of iron on macrophage production of tumoricidal free radical nitric oxide (NO) has been reported, its possible influence on macrophage anti-tumour activity has not been established. In the present study, FeSO4 markedly reduced IFN-γ + LPS-induced NO synthesis in mouse and rat macrophages. The effect of iron coincided with the loss of macrophage cytotoxic activity against NO-sensitive C6 rat astrocytoma and L929 mouse fibrosarcoma cell lines, as measured by MTT assay for cellular respiration and the crystal violet test for cell viability. Tumour cell survival did not improve further in the presence of FeSO4 if macrophage NO release and cytotoxicity were already blocked by aminoguanidine. In accordance with the results obtained with exogenous iron, cell membrane permeable iron chelator o-phenanthroline enhanced both macrophage NO release and anti-tumour activity. Iron also down-regulated NO production and increased the viability of L929 fibrosarcoma cells stimulated with IFN-γ + LPS in the absence of macrophages. However, neither NO release nor cell viability was affected by iron addition to cultures of the C6 astrocytoma cell line. Iron was unable to prevent L929 and C6 cell death induced by the NO releasing chemicals SNP and SIN-1, indicating that iron-mediated inhibition of NO synthesis, rather than interference with its cytotoxic action, was responsible for the protection of tumour cells. Collectively, these results indicate that iron might protect tumour cells by reducing both macrophage and tumour cell-derived NO release.
Influence of the route of sensitization on local and systemic immune responses in a murine model of type I allergy
Authors: REPA A , WILD C , HUFNAGL K , WINKLER B , BOHLE B , POLLAK A , WIEDERMANN U
Published: June 2004

The pathophysiological and immunological characteristics of allergic immune responses are controlled by a variety of factors. We have studied the extent to which the route of sensitization influences allergen-specific IgE synthesis and local airway inflammation using a mouse model of allergic sensitization to the major birch pollen allergen Bet v 1. Sensitization of BALB/c mice with recombinant (r)Bet v 1 was performed using intraperitoneal (IP), subcutaneous (SC) or aerosol (AS) sensitization protocols. Mice were analysed for allergen-specific serum antibodies by ELISA and IgE-dependent basophil degranulation. Proliferative responses and cytokine production of splenocytes were measured upon Bet v 1 stimulation in vitro. Bronchoalveolar lavages were performed after airway challenge with aerosolized birch pollen extract for assessment of eosinophilic airway inflammation and local cytokine production in vivo. Highest allergen specific IgE levels and IgE-dependent basophil degranulation were achieved using the SC route. High IL-5 production by spleen and lung cells was associated with pronounced eosinophilia in bronchoalveolar lavages. After IP sensitization, despite giving the highest IgG levels, only low IgE levels, basophil degranulation and IL-5 production were seen. On the other hand, AS sensitization, resulting in the lowest systemic IgE and IL-5 levels, led to a comparably strong airway inflammation as the SC route. Our finding that the route of sensitization can result in a dissociation of local and systemic immune responses may contribute to a better understanding of the pathogenesis of allergic diseases and help to develop new treatment strategies.
Oral tolerization with peptide 336–351 linked to cholera toxin B subunit in preventing relapses of uveitis in Behcet's disease
Authors: STANFORD M , WHITTALL T , BERGMEIER L A , LINDBLAD M , LUNDIN S , SHINNICK T , MIZUSHIMA Y , HOLMGREN J , LEHNER T
Published: June 2004

Behcet's disease (BD) specific peptide (p336–351) was identified within the human 60 kD heat shock protein (HSP60). Oral p336–351 induced uveitis in rats which was prevented by oral tolerization with the peptide linked to recombinant cholera toxin B subunit (CTB). This strategy was adopted in a phase I/II clinical trial by oral administration of p336–351-CTB, 3 times weekly, followed by gradual withdrawal of all immunosuppressive drugs used to control the disease in 8 patients with BD. The patients were monitored by clinical and ophthalmological examination, as well as extensive immunological investigations. Oral administration of p336–351-CTB had no adverse effect and withdrawal of the immunosuppressive drugs showed no relapse of uveitis in 5 of 8 patients or 5 of 6 selected patients who were free of disease activity prior to initiating the tolerization regimen. After tolerization was discontinued, 3 of 5 patients remained free of relapsing uveitis for 10–18 months after cessation of all treatment. Control of uveitis and extra-ocular manifestations of BD was associated with a lack of peptide-specific CD4+ T cell proliferation, a decrease in expression of TH1 type cells (CCR5, CXCR3), IFN-γ and TNF-α production, CCR7+ T cells and costimulatory molecules (CD40 and CD28), as compared with an increase in these parameters in patients in whom uveitis had relapsed. The efficacy of oral peptide-CTB tolerization will need to be confirmed in a phase III trial, but this novel strategy in humans might be applicable generally to autoimmune diseases in which specific antigens have been identified.
An experimental model for the study of transient lower oesophageal sphincter relaxation and motor function of the proximal stomach in humans
Authors: Carmagnola S. , Cantù P. , Savojardo D. , Allocca M. , Penagini R.
Published: June 2004

Abstract  A simple and reliable experimental model would be useful in human research on new drugs which target transient lower oesophageal sphincter (LOS) relaxation. The aim was to investigate the effect of repeated distensions on the rate of transient LOS relaxation, LOS pressure and motor function of the proximal stomach. Twelve healthy subjects were studied with a multilumen manometric assembly incorporating a sleeve sensor for the LOS and a bag positioned in the proximal stomach and connected to a barostat. Intrabag volume was set at 75% of the threshold for gastric discomfort and maintained for two 30‐min distension periods separated by a 45‐min washout with the bag deflated. The studies lasted 145 ± 2 min. The rate of transient LOS relaxations was similar during the two distensions, 3.5;2–4 vs 3;2.5–4 (median;interquartile range) and so was LOS pressure. Baseline intrabag pressure, as a measure of gastric tone, and the number of pressure waves, as a measure of phasic contractions, were also similar, 11.3;9.3–12.3 mmHg vs 10.8;9.3–12.5 mmHg and 16;13–28 mmHg vs 19;15–29 mmHg, respectively. Our model allows to perform 1‐day studies which can assess two experimental conditions on transient LOS relaxations and motor function of the proximal stomach within an acceptable time span.
Brain activity during distention of the descending colon in humans
Authors: Hamaguchi T. , Kano M. , Rikimaru H. , Kanazawa M. , Itoh M. , Yanai K. , Fukudo S.
Published: June 2004

Abstract  Brain‐gut interaction is considered to be a major factor in the pathophysiology of irritable bowel syndrome. However, only limited information has been provided on the influence of gastrointestinal tract stimulation on the brain. Our aim in this study was to determine the specific regions of the brain that are responsible for visceral perception and emotion provoked by distention of the descending colon in humans. Fifteen healthy males aged 22 ± 1 participated in this study. Using a colonoscope, a balloon was inserted into the descending colon of each subject. After sham stimulation, the colon was randomly stimulated with bag pressures of 20 and 40 mmHg, and regional cerebral blood flow was measured by [15O] positron emission tomography. The subjects were asked to report visceral perception and emotion using an ordinate scale of 0–10. Colonic distention pressure dependently induced visceral perception and emotion, which significantly correlated with activation of specific regions of the brain including the prefrontal, anterior cingulate, parietal cortices, insula, pons, and the cerebellum. In conclusion, distention of the descending colon induces visceral perception and emotion. These changes significantly correlate with activation of specific regions in the brain including the limbic system and the association cortex, especially the prefrontal cortex.
Effects of electroacupuncture on gastric myoelectrical activity in healthy humans
Authors: Shiotani A. , Tatewaki M. , Hoshino E. , Takahashi T.
Published: June 2004

Abstract  The acupuncture point of the wrists (PC6) and the lower legs (ST36) are common points for the treatment of gastric symptoms. However, it remains unclear whether these two acupoints have different effects on gastric myoelectrical activity. We compared the effect of electroacupuncture (EAP) between PC6 and ST36 on gastric myoelectrical activity using surface electrogastrography (EGG). EAP (1 Hz, for 30 min) was applied at either ST36, or PC6, or both acupoints in eight healthy volunteers. EAP at both PC6 and ST36 did not change the percentage of normal slow waves and tachygastria. While EAP at either PC6 or ST36 did not change period dominant frequency (PDF), EAP at both PC6 and ST36 significantly decreased PDF to 78.1 ± 8.4% of baselines. EAP at PC6 reduced period dominant power (PDP) to 47.2 ± 5.3% of baselines, while EAP at ST36 increased PDP to 153.6 ± 28.3% of baselines. EAP at shoulders (sham acupuncture) did not affect the gastric myoelectrical activity. EAP at either PC6 or ST36 shows an opposite effect on PDP, whereas EAP at both PC6 and ST36 has a synergistic effect on PDF. Understanding site‐specific effects of acupuncture may contribute to the selection of appropriate acupoints for treating functional GI disorders.
Motor responses of rat hypertrophic intestine following chronic obstruction
Authors: Bertoni S. , Gabella G. , Ghizzardi P. , Ballabeni V. , Impicciatore M. , Lagrasta C. , Arcari M. L. , Barocelli E.
Published: June 2004

Abstract  The present work aims at investigating the changes in motor responsiveness of rat intestine hypertrophied by chronic mechanical obstruction. Motor responses to pharmacological agents and electrical field stimulation (EFS) were studied in hypertrophic ileal segments excised from rats subjected to experimental stenosis (n = 20) and compared with responses of control tissues from sham‐operated animals (n = 20). Spontaneous motility and contractile responses to exogenous agents (KCl, acetylcholine and substance P) and EFS (10‐s trains every minute, 120 mA, 0.5 ms, 1–10 Hz) were increased in hypertrophic longitudinal segments; however, normalization of motor responses to tissue wet weight revealed a remarkable reduction of contractile efficiency in hypertrophied tissues coupled with a loss of sensitivity to nitric oxide‐mediated relaxation. Furthermore, EFS under non‐adrenergic non‐cholinergic (NANC) conditions unveiled a major role of the cholinergic component over the peptidergic one in the neurogenic contraction of hypertrophic intestine. On the whole, hypertrophic intestinal growth emerges as a dynamic process entailing adaptation of smooth muscle and neuronal structures to the increased functional load imposed by lumen obstruction.
Intolerance to visceral distension in functional dyspepsia or irritable bowel syndrome: an organ specific defect or a pan intestinal dysregulation?
Authors: Bouin M. , Lupien F. , Riberdy M. , Boivin M. , Plourde V. , Poitras P.
Published: June 2004

Abstract  Functional gastrointestinal disorders (FGID) are characterized by visceral hypersensitivity that could be specific to a region of the gut or reflect a diffuse pan‐intestinal disorder. Sensory thresholds to distension at two visceral sites in patients with different FGIDs were determined. According to Rome II criteria, 30 patients from three groups were studied: patients with (i) functional dyspepsia (FD) or (ii) irritable bowel syndrome (IBS), and (iii) patients with concomitant symptoms of FD and IBS. Pain thresholds to balloon distension were determined in stomach and rectum. In FD patients, gastric intolerance to balloon distension was found in 91% patients; rectal hypersensitivity was documented in 18% patients. In IBS patients, rectal hypersensitivity was seen in 75% patients; while gastric hypersensitivity was never found. In patients with concomitant symptoms of FD + IBS, gastric and rectal intolerance to distension were present respectively in 82 and 91% patients. In the whole group, visceral intolerance to distension was documented at one site in 90% patients and at both sites, i.e. stomach and rectum, in 33% patients. Visceral intolerance to distension can be pan‐intestinal in patients with multiple sites of symptoms, but appears organ‐specific in patients exhibiting a specific site of symptoms.
Analysis of surface geometry of the human stomach using real‐time 3‐D ultrasonography in vivo
Authors: Liao D. , Gregersen H. , Hausken T. , Gilja O. H. , Mundt M. , Kassab G.
Published: June 2004

Abstract  The objective of this study was to develop an analytical method to describe the three‐dimensional (3‐D) geometry of the gastric antrum, gastric fundus and the whole stomach. The Fourier series method was used to simulate the organ surface geometry obtained from a 3‐D ultrasound system. Data generated from eight antrums and three whole stomachs, at pressures of approximately 7 cmH2O, were used for lumen curvature calculations. The principal curvatures spatial distributions were non‐homogeneous in the gastric antrum, gastric fundus and the stomach due to their complex geometry. The maximum longitudinal principal curvature in the antrum, fundus and total stomach were, respectively, 0.460 ± 0.066, 0.583 ± 0.087 and 1.123 ± 0.328, whereas the maximum circumferential curvature were 1.192 ± 0.090, 3.649 ± 1.574 and 8.444 ± 3.424, respectively. The present study provides an analytical tool for characterizing the complex 3‐D geometry of an organ‐like the human stomach reconstructed by clinical imaging modalities. Providing an average tension for the stomach does not reflect the large variation in tension throughout the stomach wall.
Diabetic autonomic neuropathy: evidence for apoptosis in situ in the rat
Authors: Guo C. , Quobatari A. , Shangguan Y. , Hong S. , Wiley J. W. , Quobatari A.
Published: June 2004

Abstract  We examined the hypothesis that activation of the apoptosis cascade occurs relatively early in diabetes mellitus affecting three distinct neuronal populations that are involved in regulating gut function: (i) dorsal root ganglion (DRG), (ii) vagus nodose ganglion and (iii) colon myenteric plexus. A validated streptozotocin‐induced diabetic rat model and age‐matched healthy controls were studied. After 4–8 weeks of diabetes the animals were anaesthetized, fixed in situ and the relevant tissues removed. After 1 month of diabetes some animals were treated with insulin for 2 weeks to restore euglycaemia. Apoptosis was measured using immunohistochemical detection of activated caspase‐3 and terminal deoxynucleotidyl transferase‐mediated dUTP nick‐end labelling (TUNEL)‐positive cells in adjacent sections in neurones (PGP 9.5‐positive cells). The level of apoptosis was confirmed using double‐label assessment of caspase‐3 and TUNEL in DRG preparations. Caspase‐3 immunoreactive neurones demonstrated a range in staining intensity. When all grades of staining were included, 6–8% of the DRG, nodose ganglia and myenteric neurones were immunoreactive in the preparations from diabetic rats compared with 0.2–0.5% in controls. Neurones staining positive for both caspase‐3 and TUNEL accounted for 1–2% of the total neuronal population in all three preparations in diabetic rats compared with 0.1–0.2% in controls (P < 0.05). Insulin treatment reversed the percentage of TUNEL‐positive neurones in diabetic rats to control levels. Activation of the apoptosis cascade occurs relatively early in diabetic autonomic neuropathy and may contribute to the pathophysiology of this disorder.
Microinjection of glutamate into dorsal motor nucleus of the vagus excites gallbladder motility through NMDA receptor – nitric oxide – cGMP pathway
Authors: Liu C. Y. , Xie D. P. , Liu J. Z.
Published: June 2004

Abstract  We have reported that both glutamate and nitric oxide (NO) participated in the regulation of gallbladder motility in dorsal motor nucleus of the vagus (DMV). The aim of this study is to investigate the type of receptor in DMV that mediates the excitatory effect of glutamate on gallbladder motility and the correlation between the glutamate and NO. A frog bladder connected with a force transducer was inserted into the gallbladder to record the change of gallbladder pressure. Glutamate (65 mmol L−1, 100 nL) microinjected into DMV significantly increased the strength of gallbladder phasic contraction. This effect was abolished by ketamine (180 mmol L−1, 100 nL), the specific N‐methyl‐d‐aspartic acid (NMDA) receptor antagonist, but was not influenced by 6‐cyaon‐7‐nitroquinoxaline‐2,3‐(1H,4H)‐dione (CNQX) (180 mmol L−1, 100 nL), the non‐NMDA ionotropic receptor antagonist. NG‐nitro‐l‐arginine‐emthyl (l‐NAME) (1 mol L−1, 100 nL), the nitric oxide synthase (NOS) inhibitor, reversed the excitatory effect of glutamate on gallbladder motility. Microinjection of sodium nitroprusside (SNP), the NO donor, into DMV enhanced the gallbladder motility, and this effect was not modulated by ketamine. Microinjection of NMDA (5 mmol L−1, 100 nL) increased the strength of gallbladder phasic contraction, and this effect was attenuated by methylene blue (100 mmol L−1, 100 nL), the soluble guanylate cyclase inhibitor. These results suggest that glutamate regulate the gallbladder motility through the NMDA receptor – NO – cGMP pathway in DMV.
Gastric cancer with metastasis to the gingiva
Authors: SHIMOYAMA SHOUJI , SETO YASUYUKI , AOKI FUMIO , OGAWA TOSHIHISA , TOMA TOSHIYUKI , ENDO HISAKO , ITOUJI TORU , KAMINISHI MICHIO
Published: June 2004

The present case report describes a gastric cancer which showed unusual metastasis in the oral region. A 56‐year‐old male patient underwent total gastrectomy and splenectomy due to advanced gastric cancer in the upper third of the stomach. Fifteen months later, he presented with anorexia and gingival swelling of durations of approximately 3 and 1 month, respectively. The gastric tumor was histologically a signet ring cell and a poorly differentiated cancer with a moderate degree of vascular invasion. Biopsy specimens from the gingival tumor revealed a signet ring cell cancer. Other metastatic sites were the brain, limb bones and abdominal lymph nodes. A bone scintigram revealed an abnormal uptake in the limb bones, while it did not exhibit any abnormality in the oral region. Correlation between the histology of the gingival tumor with that of the gastric cancer, as well as the absence of a gingival tumor at the time of prior gastrectomy, led to a diagnosis that the gingival tumor was a metastasis from gastric cancer. Gastric cancer metastasizing to the oral region, either the osseus or the oral soft tissue, is very rare. Although it cannot be proved without an autopsy, negative findings in the mandible by bone scanning in the present case suggest that direct gingival metastasis can be considered, rather than mandibular metastasis involving the gingiva. Hematogenous spread could be a mechanism of metastasis for this unusual tumor.
Taurocholate transport by hepatic and intestinal bile acid transporters is independent of FIC1 overexpression in Madin–Darby canine kidney cells
Authors: HARRIS MATTHEW J , KAGAWA TATEHIRO , DAWSON PAUL A , ARIAS IRWIN M
Published: June 2004

Background:  Mutations in the human familial intrahepatic cholestasis gene, FIC1, result in progressive familial intrahepatic cholestasis type 1 in children and benign recurrent intrahepatic cholestasis. The present study was performed to determine whether FIC1 transports bile acids and/or influences the activity of apical bile acid transporters.

Methods:  The apical secretion assay utilized transfected Madin–Darby canine kidney (MDCK) cells, which stably express the bile acid uptake protein, Na+/taurocholate co‐transporting polypeptide (NTCP). These cells were then transiently transfected with FIC1 and/or the bile salt export pump (BSEP) and were grown on Transwell filters to form a polarized monolayer. [3H]Taurocholate was added to the basal medium and the taurocholate secretion was measured in the apical medium. A second assay, apical uptake assays, utilized polarized MDCK‐II cells, which were transiently transfected with FIC1, FIC1 mutants and/or the apical sodium‐dependent bile acid transporter (ASBT). [3H]Taurocholate was added to the apical media and intracellular uptake of taurocholate was measured.

Results:  Apical secretion assays: FIC1 expression in MDCK/NTCP cells had no effect on taurocholate secretion compared to controls. In contrast, apical secretion of taurocholate in BSEP‐transfected cells was approximately twofold higher than in non‐transfected MDCK/NTCP cells (P < 0.01). The BSEP‐mediated secretion was unaffected by co‐transfection with FIC1. Apical uptake assays: taurocholate uptake in ASBT expressing cells was 6.5‐fold higher than in controls and was unaffected by co‐transfection of cells with FIC1 or FIC1 mutants.

Conclusions:  These results indicate that FIC1 does not transport taurocholate and, when overexpressed in MDCK cells, had no effect on the function of BSEP or ABST.
Reduced expression of peroxisome proliferator‐activated receptor‐α may have an important role in the development of non‐alcoholic fatty liver disease
Authors: YEON JONG EUN , CHOI KYUNG MOOK , BAIK SEI HYUN , KIM KYOUNG OH , LIM HYOUNG JOON , PARK KI HO , KIM JIN YONG , PARK JONG‐JAE , KIM JAE SEON , BAK YOUNG‐TAE , BYUN KWAN SOO , LEE CHANG HONG
Published: June 2004

Background:  Although the pathogenesis of non‐alcoholic fatty liver disease (NAFLD) remains poorly understood, metabolic syndrome associated with insulin resistance is the most reproducible factor in the development of NAFLD. Fat accumulation in hepatocytes results from an imbalance in the input, output and oxidation of fatty acid. Peroxisomes contain a battery of fatty acid oxidizing enzymes, the first of which, acyl‐CoA oxidase (AOX), initiates the β‐oxidation spiral. One of the mammalian peroxisome proliferator‐activated receptors (PPAR), PPAR‐α, regulates the transcriptional expression of the enzymes involved in fatty acid β‐oxidation. The aim of the present study was to define the role of PPAR‐α and AOX in the development of NAFLD using the Otsuka Long‐Evans Tokushima fatty (OLETF) rat model.

Methods:  Liver tissue from OLETF (n = 12) and control (n = 10) rats 12, 28, and 40 weeks old were processed for histopathological and western blot analysis. The messenger RNA of PPAR‐α and AOX were quantified by real‐time RT‐PCR.

Results:  At 40 weeks old, the histological analysis of the OLETF rat liver had steatosis (approximately 66%) and mild inflammation, which were comparable to those in NAFLD. Histological changes were unremarkable  in  12 week  and  28 week  rats.  In  12 week  OLETF  rats,  the  mRNA  of  AOX  was  63% of the control. Expression of PPAR‐α mRNA was also reduced to 3% that of the control. Along with the changes of mRNA, the protein expression of PPAR‐α was also significantly reduced to 17% that of the control. In 28 week and 40 week animals, PPAR‐α protein expression gradually increased to 75% and 78% that of the control. Expression of PPAR‐α mRNA was also increased by up to 26% and 110% of the control. AOX, regulated by PPAR‐α, also increased to 149% and 120% of the control.

Conclusion:  Reduced expression of PPAR‐α and AOX was observed even before definite steatosis had developed. The alteration of peroxisomal fatty acid metabolism may have an important role in the development of NAFLD.
Preliminary analysis of a newly proposed prognostic scoring system (SLiDe score) for hepatocellular carcinoma
Authors: OMAGARI KATSUHISA , HONDA SUMIHISA , KADOKAWA YOSHIKO , ISOMOTO HAJIME , TAKESHIMA FUMINAO , HAYASHIDA KENJI , MIZUTA YOHEI , MURATA IKUO , KOHNO SHIGERU
Published: June 2004

Background:  The long‐term prognosis of hepatocellular carcinoma (HCC) remains poor and the prediction of survival is often difficult because of the limited liver function and frequent recurrence of HCC in most patients. Therefore, a prognostic classification of HCC should account for both tumor‐related variables and liver function.

Methods:  The value of reported prognostic factors for HCC was assessed and a new prognostic classification was established called the ‘SLiDe’ scoring system (S, stage; Li, liver damage; De, des‐γ‐carboxy prothrombin) using ‘stage’ and ‘liver damage’ of the recently revised 4th edition of the Japanese staging system edited by the Liver Cancer Study Group of Japan, and the serum level of des‐γ‐carboxy prothrombin (DCP) in 177 patients with HCC.

Results:  Univariate analysis identified Child–Pugh stage, liver damage, tumor morphology, portal vein thrombosis, stage, serum level of α‐fetoprotein (AFP), serum level of DCP, and initial treatment as significant prognostic factors. Of these, liver damage, stage, and serum level of DCP remained independent predictive factors of survival after multivariate prognostic analysis using the proportional hazards regression model. Therefore, a new prognostic scoring system (SLiDe scoring system) was derived that assigned a linear score (0/1/2/3) to these three covariates. This SLiDe scoring system was statistically a better model for predicting outcome in the present study population than the Cancer of the Liver Italian Program (CLIP) and the Japan Integrated Staging (JIS) scoring systems, as judged by the Akaike Information Criteria.

Conclusion:  The SLiDe scoring system is useful for the assessment of the prognosis of patients with HCC as long as the Japanese staging system is used, although this uses parameters such as the indocyanine green retention test and DCP, which are not examined routinely in every part of the world. Therefore, the proposed classification should be further validated in other large study populations.

© 2004 Blackwell Publishing Asia Pty Ltd
Efficacy of transanal endoscopic microsurgery in the management of rectal polyps
Authors: PLATELL CAMERON , DENHOLM EVA , MAKIN GREGORY
Published: June 2004

Background and Aim:  Transanal endoscopic microsurgery (TEM) was a technique developed to allow the excision of adenomas and early‐stage cancers from the rectum and distal sigmoid colon. The aim of this project was to prospectively study surgical morbidity, mortality and the local recurrence rate of all patients treated with this technique.

Methods:  All patients undergoing TEM were prospectively evaluated. Endpoints to assess the surgical morbidity and mortality were defined before the study commenced. All patients underwent regular follow up to determine treatment efficacy in terms of the local recurrence rate and survival.

Results:  The study involved 113 patients, with a mean age of 69 years (standard deviation 14 years, range 30–94 years), and a male to female ratio of 1.4:1. The mean polyp area was 20.5 cm2 (range 1–169 cm2) and the mean height above the anal verge was 9.5 cm (range 4–25 cm). Histology of the tumors found 62 adenomas, 20 carcinomas in situ, and 31 adenocarcinomas. There were no unplanned returns to theater or postoperative deaths. Four patients required readmission within 30 days because of bleeding, and nine patients underwent more radical surgical procedures following histological evaluation of the resected specimens. During a mean follow up of 1.5 ± 0.8 years (maximum 3.2 years), there have been two recurrences of villous adenomas. The actuarial local recurrence rate at 2 years is 2.4% (95% confidence interval 0.8–4.0%).

Conclusion:  TEM was demonstrated to be a safe surgical procedure, and early follow up has shown it to be an efficacious treatment for benign rectal adenomas and early rectal cancers.

© 2004 Blackwell Publishing Asia Pty Ltd
Chromosomal and microsatellite instability in sporadic gastric cancer
Authors: HIYAMA TORU , TANAKA SHINJI , YOSHIHARA MASAHARU , SASAO SHOGO , KOSE KAZUHIRO , SHIMA HIDEYUKI , TUNCEL HANDAN , UENO YOSHITAKA , ITO MASANORI , KITADAI YASUHIKO , YASUI WATARU , HARUMA KEN , CHAYAMA KAZUAKI
Published: June 2004

Background:  Gastric cancer can progress through two pathways of genomic instability: chromosomal (CIN) and microsatellite instability (MSI). It is hypothesized that these two pathways are not always independent and that some tumors show overlap between these two mechanisms.

Methods:  A total of 98 sporadic gastric cancers were classified based on their MSI status, using microsatellite assay with BAT26. Evidence for CIN was investigated by identifying loss of heterozygosity (LOH) events on chromosome arms, 5q, 10p, 17p, 17q, and 18q, which are regions harboring tumor suppressor genes that are significant in gastric cancer development.

Results:  Twelve tumors (12%) showed high‐frequency MSI (MSI‐H). Overall, 43 of the tumors (44%) had at least one LOH event, with most frequent chromosomal losses observed on 10p and 18q (30%, respectively), followed by 5q (21%), 17p (14%), and 17q (12%). Interestingly, overlap was observed between CIN and MSI pathways. Of 43 cancers with LOH events, four (9%) were also MSI‐H. It was also found that 48% of cancers without MSI‐H had no LOH events identified, comprising a subgroup of tumors that were not representative of either of these two pathways of genomic instability.

Conclusion:  These results suggest that molecular mechanisms of genomic instability are not necessarily independent and may not be fully defined by either the MSI or CIN pathways in sporadic gastric cancers.
Cisapride treatment of constipation–predominant irritable bowel syndrome is not superior to placebo
Authors: ZIEGENHAGEN DIETER J , KRUIS WOLFGANG
Published: June 2004

Background and Aim:  Previous studies with cisapride reported conflicting results in patients with constipation–predominant irritable bowel syndrome (IBS). To gain further evidence, this randomized double‐blind study was carried out.

Methods:  Eighty‐two symptomatic outpatients were randomized to receive either 5 mg oral cisapride or placebo three times daily for a period of 12 weeks. In patients without satisfactory improvement after 4 weeks, the dose was doubled. Symptom evaluation used visual analog scales (VAS) and the investigators’ global assessment.

Results:  After 4 weeks, in 18 (45%) cisapride and 24 (57%) placebo patients the dose was doubled because of insufficient improvement of symptoms. The mean VAS score for patients’ global rating of IBS symptoms at baseline was 67.5 mm for cisapride versus 70.7 mm for placebo, and improved to 38.4 mm versus 44.5 mm after 12 weeks of treatment. Investigators rated the overall effect of therapy as good or excellent in 70% of the cisapride and 50% of the placebo group. Neither these nor further efficacy parameter differences reached statistical significance.

Conclusions:  These results indicate that the effect of 15–30 mg cisapride daily on symptoms of constipation‐predominant IBS is not significantly superior to placebo. During the 12 week treatment of this trial cisapride proved to be safe and tolerable.
Differentiation of cystic pancreatic neoplasms and pseudocysts by conventional and echo‐enhanced ultrasound
Authors: RICKES STEFFEN , WERMKE WOLFRAM
Published: June 2004

Background and Aims:  Echo‐enhanced sonography is a non‐invasive and increasingly used procedure for the differentiation of pancreatic tumors. However, the diagnostic accuracy of this procedure compared to conventional ultrasound for the differential diagnosis of cystic pancreatic neoplasms from pseudocysts has never been investigated in a prospective study.

Methods:  Thirty‐one patients with a cystic pancreatic lesion at the conventional ultrasound (mean age 57 years, range 36–82 years) were included in the study. Sonography was performed by an experienced examiner who was unaware of the patients’ clinical diagnosis. The exact diagnosis was based on histological evidence from biopsy examination (surgical or transabdominal fine needle biopsy for all cystic neoplasms and five pseudocysts), or a follow‐up of at least 18 months (four pseudocysts).

Results:  Of the 31 patients, 19 had cystadenomas, three had cystadenocarcinomas, and nine had pseudocysts. Only 27% of the cystadenomas and 67% of the pseudocysts could be correctly classified by conventional ultrasound. Conversely, 95% of the cystadenomas (P = 0.0001) and all pseudocysts were diagnosed correctly by echo‐enhanced sonography. The sensitivity of echo‐enhanced sonography with respect to diagnosing cystadenoma was 95% and its specificity was 92%. The corresponding values for pseudocysts were both 100%.

Conclusion:  Echo‐enhanced sonography has a high sensitivity and specificity in the differential diagnosis of cystic pancreatic tumors. With this procedure the differentiation of cystadenomas and pseudocysts can be improved. However, histology is the standard of reference.
Hepatitis E virus infection may be transmitted through blood transfusions in an endemic area
Authors: KHUROO MOHAMMAD SULTAN , KAMILI SALEEM , YATTOO GHULAM NABI
Published: June 2004

Aim:  To address the issue of whether or not hepatitis E virus (HEV) is transmitted parenterally.

Methods:  We conducted a retrospective study which involved 145 multiple transfused patients and 250 healthy controls. A prospective study was also undertaken involving 50 hospitalized patients, 25 of whom were transfused with 107 blood units, while the other 25 did not receive any transfusions.

Results:  In our retrospective study, markers of acute HEV infection (IgM anti‐HEV and HEV RNA) were detected in a significantly higher number of multiple transfused patients (13 of 145) compared to controls (two of 250) (P < 0.001; OR = 12.21 [95% confidence interval: 2.71–54.70]). All 13 HEV‐infected patients had been transfused at least once in a 3‐month period before testing. Overall, patients positive for any of the HEV markers (IgG, IgM or HEV RNA) had received more blood transfusions, had higher occurrence of icteric disease and higher serum alanine aminotransferase levels. In our prospective study, IgG anti‐HEV was detected in 11 of 107 donor samples, three of 25 patients in their pretransfusion samples (one sample was positive for IgM anti‐HEV as well) and two of 25 control patients. Post‐transfusion HEV infection developed in three of 22 susceptible (IgG anti‐HEV negative) transfused patients; the infection was traced to their four respective donors who were asymptomatic, HEV RNA positive (4/4) and IgM anti‐HEV positive (3/4). In contrast, none of the non‐transfused patients developed HEV infection during the follow‐up period.

Conclusion:  Frequent transmission of HEV by blood transfusion places recipients at risk and warrants redefining of the donor screening policy by blood banks, especially in endemic areas.
Internal loading: A new solution to an old problem in aquatic sciences
Authors: Håkanson Lars
Published: June 2004

Internal loading has long been regarded as an ‘Achilles heel’ in aquatic science and management. Internal loading is of fundamental importance in large and shallow lakes, where even low wind velocities can cause a considerable resuspension of matter deposited on the lake bed. The resuspended matter, and the chemical substances bound to the resuspended matter, will influence almost all processes in the aquatic ecosystem, such as water clarity and depth of the photic zone, and hence, primary and secondary production. If the sediments are contaminated, it will increase the concentrations of harmful substances in water and sediments and the potential ecosystem effects related to such concentrations. This paper presents an overview of the processes regulating bottom dynamic conditions in lakes (erosion, transport, accumulation), provides examples on the role of internal loading within the context of limnology and water management, and presents a new, general approach to quantify internal loading from sediments in lakes. The new approach has been critically tested, being a key factor behind the increase in predictive power of a new generation of lake models meant to be used for practical water management. Internal loading of any water pollutant depends on sedimentation. Sedimentation in this approach is presented as a function of two substance‐specific variables, including the fall velocity of the carrier‐particles and the particulate fraction (which, by definition, is the only fraction of a water pollutant that can settle out on the lake bed), and three generic variables, including mean depth, suspended particulate matter and ET‐areas (areas of erosion and transport). On ET‐areas there is, by definition, a discontinuous sedimentation of materials that settles according to Stokes' law. Basically, internal loading is the sum of advective (resuspension) and diffusive transport from the sediments. Resuspension from ET‐areas is given as a function of the lake form (a new algorithm based on the volume development) and the age of ET‐sediments.
Sediment dynamics and pollutant mobility in rivers: An interdisciplinary approach
Authors: Förstner Ulrich
Published: June 2004

Characteristic dynamic features of sediment‐related processes in rivers include dramatic effects of stormwater events on particle transport, rapid and far‐reaching effects of sulphide oxidation during resuspension, and biological accumulation and potential release of toxic chemicals. Pollutant mobility is the net result of the stabilizing and mobilizing effects in both hydraulic and chemical fields. In practice, emphasis has to be given to fine‐grained sediments and suspended matter as these materials exhibit large surface areas and high sorption capacities. Organic materials are highly reactive. Degradation of organic matter will induce oxygen depletion and might enhance formation of flocs and biofilms. Study of variations of sediment and water chemistry should predominantly include changes of pH and redox conditions, competition of dissolved ions and processes such as complexation by organic substances. Major questions relate to the potential reduction of sorption sites on minerals and degradation of organic carrier materials. All these processes will influence solution/solid equilibrium conditions and have to be studied prior to modelling the overall effects of pollutants on the water body and aquatic ecosystems. With respect to handling and remediation of contaminated river sediments, either in‐place or excavated, a chemical and biological characterization of the material, of the (disposal) site and of the long‐term processes is crucial. Passive techniques (e.g. in situ stabilization, subaqueous capping) provide economic advantages as there are no operation costs following their installation. However, the success of these ecological and geochemical engineering approaches is mainly based on an in‐depth knowledge of the underlying processes.