bluuebunny/crossref_metadata_2025 huggingface dataset:
Enalaprilat, losartan and LU 135252 in coronary blood flow regulation
Authors: Rigol M. , Heras M. , Solanes N. , Epelde F. , Roig E. , Pérez‐Villa F. , Roqué M. , Sanz G.
Published: December 2004

Background High plasma levels of angiotensin II are found in several pathologies such as hypertension, heart failure and myocardial infarction. The effect of high concentrations of angiotensin II on coronary circulation is not well defined. The aim of the present study was to assess coronary blood flow regulation during tachycardia in the presence of elevated coronary plasma levels of angiotensin II, and the changes induced by ACE inhibition and blockade of angiotensin II and endothelin‐A receptors.

Design Left anterior coronary artery was catheterized in 38 pigs to infuse the study drugs. Saline was infused for 15 min. Then, the first atrial pacing was performed. The pigs were distributed to: Group 1 (n = 7) angiotensin II; Group 2 (n = 7) enalaprilat + angiotensin II; Group 3 (n = 9) the bradykinin B2 antagonist HOE 140 + enalaprilat + angiotensin II; Group 4 (n = 7) losartan + angiotensin II; and Group 5 (n = 8) endothelin‐A receptor antagonist LU 135252 + angiotensin II. After giving these infusions, a second pacing was repeated.

Results The increase in coronary blood flow induced by pacing with angiotensin II was reduced from 181 ± 21% to 116 ± 37% (P = 0·006 vs. saline). Enalaprilat, losartan and LU 135252 restored the capacity of coronary blood flow to increase during pacing (151 ± 39%, 162 ± 35% and 161 ± 16%, respectively; P = NS, vs. saline), while HOE 140 abolished the effect of enalaprilat.

Conclusions Moderately elevated coronary concentrations of angiotensin II reduced coronary blood flow during pacing. Enalaprilat, losartan and LU 135252 restored the hyperaemic coronary flow to similar values observed with saline. The beneficial effect of ACE inhibition is mediated through an increase in bradykinin.
Increased intestinal permeability as a cause of fluctuating postprandial blood glucose levels in Type 1 diabetic patients
Authors: Damci T. , Nuhoglu I. , Devranoglu G. , Osar Z. , Demir M. , Ilkova H.
Published: December 2004

Background In diabetic patients, postprandial glucose levels, which have a major impact on metabolic control, are determined by the rate of nutrient delivery into the intestine, absorption of nutrients from the small intestine, and the metabolism of the absorbed nutrients by the liver. The present study addresses whether Type 1 diabetic patients have increased intestinal permeability and intestinal permeability predicts postprandial glucose variability.

Material and methods Thirty Type 1 diabetic patients together with 15 sex‐ and age‐matched healthy controls were enrolled in the study. After an overnight fasting all patients and controls received 100 µCi 51Cr of EDTA as a radioactive tracer and the percentage of the isotope excreted in a 24‐h urinary specimen was the permeability measure. Instant blood glucose was measured just before the test, and the patients performed and recorded self‐monitoring of fasting and 2nd‐hour postprandial blood glucose levels during the following week.

Results We found that intestinal permeability is increased in Type 1 diabetic patients compared with age‐ and sex‐matched healthy controls. Increased intestinal permeability is related at least in part to the instant blood glucose level and the presence of diabetic autonomic neuropathy.

Conclusion Increased intestinal permeability leads to higher variation in postprandial blood glucose levels, thereby worsening metabolic control.
The variability of liver graft function and urinary 6β‐hydroxycortisol to cortisol ratio during liver regeneration in liver transplant recipients
Authors: Kishino Satoshi , Ogawa Maki , Takekuma Yoh , Sugawara Mitsuru , Shimamura Tsuyoshi , Furukawa Hiroyuki , Todo Satoru , Miyazaki Katsumi
Published: December 2004

Abstract:  The urinary ratio of 6β‐hydroxycortisol to cortisol (6β‐OHF/F) is considered to be the simplest and most practical method for estimation of hepatic cytochrome P450 3A4 (CYP3A4) activity as a non‐invasive marker of human in vivo CYP3A4 activity. However, the inter‐ and intra‐individual variability of the urinary 6β‐OHF/F ratio during liver regeneration and the effect of variability on optimal dose of tacrolimus have not yet been clarified. The objective of this study was to clarify the change in the urinary 6β‐OHF/F ratio during liver regeneration and to determine the effect of the liver graft function on the optimal tacrolimus dose in liver transplant recipients. Two liver transplant recipients (one male and one female) and eight healthy volunteers (five males and three females) were enrolled in this study. Urine samples were collected from the recipients from 08.00 hours for 24 h on post‐transplant period, 1–10 and 21–30 days postoperatively. In the healthy volunteers, morning spot urine samples were collected at 08.00 hours. The mean urinary 6β‐OHF/F ratio in the immediate postoperative period was significantly low (p < 0.05). However, a marked difference in the regulation of CYP3A4 activity during liver regeneration was found in the two recipients. A significant correlation was found between the urinary 6β‐OHF/F ratio and the C/D ratio of tacrolimus (R = 0.658, p < 0.05). The urinary 6β‐OHF/F ratio is a useful probe for estimating the variability of CYP3A4 activity in liver transplant recipients in early postoperative phase. Future studies should evaluate the clinical usefulness of the urinary 6β‐OHF/F ratio as a predictor of tacrolimus pharmacokinetics in liver transplantation.
Relationship between endomyocardial biopsy score and cyclosporine 2‐h post‐dose levels (C2) in heart transplant patients receiving anti‐thymocyte globulin induction
Authors: Cantarovich Marcelo , Giannetti Nadia , Cecere Renzo
Published: December 2004

Abstract: Background: Cyclosporine (CsA) 2‐h post‐dose levels (C2) correlate better with the area‐under‐the‐curve compared with trough levels. The purpose of this study was to determine the relationship between C2and endomyocardial biopsy (EMB) score in heart transplant patients receiving anti‐thymocyte globulin (ATG)‐induction.

Methods: We reviewed 517 EMB performed during the first year in 39 adult heart transplant patients receiving ATG‐induction, corticosteroids, mycophenolate mofetil and CsA. C2obtained on the day of EMB was related to the International Society for Heart and Lung Transplantation classification (score ≤2 or ≥3A). Furthermore, EMB were related to C2(ng/mL), sorted according to the lower recommended range in liver (0–3 months: 850; 4–6 months: 700; 7–12 months: 500) or renal transplantation (0–1 month: 1500; 2–3 months: 1200; 4–6 months: 1000; 7–12 months: 800).

Results: Overall, C2did not significantly differ in patients with EMB ≤2 or ≥3A. However, during the first month, EMB ≤2 was associated with a trend towards a higher mean C2compared with EMB ≥3A (750 ng/mL vs. 530 ng/mL). When C2were sorted according to the lower recommended range in liver or renal transplantation, no significant difference was observed when EMB was ≤2 or ≥3A.

Conclusion: In heart transplant patients receiving ATG‐induction, C2did not significantly differ according to EMB ≤2 or ≥3A. No significant relationship was found between EMB score and C2based on the lower recommended range in liver or renal transplantation. However, mean C2>750 ng/mL appears to be associated with a lower rejection score during the first month.
Basiliximab in association with tacrolimus and steroids in caucasian cadaveric renal transplanted patients: significant decrease in early acute rejection rate and hospitalization time
Authors: Leonardi Gianluca , Messina Maria , Giraudi Roberta , Pellu Valentina , Fop Fabrizio , Segoloni Giuseppe Paolo
Published: December 2004

Abstract:  Safety and tolerability of basiliximab in renal transplantation have been proven in different immunosuppressive regimens. Few informations are available about the association of basiliximab with tacrolimus and steroids. We present a retrospective analysis performed in Caucasian cadaveric renal transplant recipients, comparing a basiliximab, tacrolimus and steroids induction protocol (GrA: 51 patients) with a tacrolimus and steroids protocol (GrB: 46 patients). A significant decrease in acute rejection rate in the first 3 months (2.0% vs. 17.4%; p < 0.01) was noted. Interestingly, the recipients in GrA were at major immunologic risk for the younger age of recipients, the greater number of mismatches and the higher rate of second transplants. The hospitalization times resulted reduced of 5.3 d in GrA vs. GrB (20.8 d vs. 26.1 d; p < 0.05). The adverse events patterns and profiles were similar in the two treatments groups. One patient in each group had a post‐transplant lymphoprolipherative disorder. No significant difference was found in patient and graft survival. According to the results of this study, in a Caucasian adult population, basiliximab in association with tacrolimus and steroids is a safe and efficacious tool for acute rejection prevention and it is cost saving by reducing the hospitalization times.
Delay of CMV infection in high‐risk CMV mismatch lung transplant recipients due to prophylaxis with oral ganciclovir
Authors: Palmer Scott M , Grinnan Dan C , Diane Reams B , Steele Mark P , Messier Robert H , Duane Davis R
Published: December 2004

Abstract: Cytomegalovirus (CMV) is a common opportunistic infection in lung transplant recipients. Despite the use of early post‐operative intravenous ganciclovir, most high‐risk patients develop CMV infection. We conducted this retrospective study to determine the efficacy of extended CMV prophylaxis with oral ganciclovir in high‐risk, donor‐positive–recipient‐negative, lung recipients. All patients initially received 3 months of intravenous ganciclovir and CMV hyperimmune globulin. Clinical outcomes in all CMV mismatch patients undergoing lung transplant surviving at least 3 months were included (n = 42). Since 1998, 14 patients received no oral ganciclovir prophylaxis (group 1) and 28 patients received indefinite oral ganciclovir after completion of intravenous therapy (group 2). In those patients receiving oral ganciclovir, the prevalence of post‐transplant CMV infection was significantly reduced over the first 180 d post‐transplant (50% in group 1 vs. 4% in group 2; p < 0.001). Although some CMV events were observed with additional follow‐up in group 2, there remained a significantly greater freedom from CMV infection by Kaplan–Meier analysis in group 2 as compared with group 1, with over 30 months follow‐up time in each group (log‐rank, p = 0.02). A moderate rate of drug discontinuation was observed in group 2, and no severe drug‐related events occurred. In high‐risk lung transplant recipients, CMV prophylaxis with intravenous ganciclovir, followed by indefinite oral ganciclovir, significantly delays and reduces post‐transplant CMV infections. A larger prospective randomized study is needed to confirm the benefits of oral ganciclovir on CMV prevention.
Mitochondrial β‐oxidation
Authors: Bartlett Kim , Eaton Simon
Published: December 2004

Mitochondrial β‐oxidation is a complex pathway involving, in the case of saturated straight chain fatty acids of even carbon number, at least 16 proteins which are organized into two functional subdomains; one associated with the inner face of the inner mitochondrial membrane and the other in the matrix. Overall, the pathway is subject to intramitochondrial control at multiple sites. However, at least in the liver, carnitine palmitoyl transferase I exerts approximately 80% of control over pathway flux under normal conditions. Clearly, when one or more enzyme activities are attenuated because of a mutation, the major site of flux control will change.
Acyl‐CoA dehydrogenases and acyl‐CoA oxidases
Authors: Kim Jung‐Ja P. , Miura Retsu
Published: December 2004

Acyl‐CoA dehydrogenases and acyl‐CoA oxidases are two closely related FAD‐containing enzyme families that are present in mitochondria and peroxisomes, respectively. They catalyze the dehydrogenation of acyl‐CoA thioesters to the corresponding trans‐2‐enoyl‐CoA. This review examines the structure of medium chain acyl‐CoA dehydrogenase, as a representative of the dehydrogenase family, with respect to the catalytic mechanism and its broad chain length specificity. Comparing the structures of four other acyl‐CoA dehydrogenases provides further insights into the structural basis for the substrate specificity of each of these enzymes. In addition, the structure of peroxisomal acyl‐CoA oxidase II from rat liver is compared to that of medium chain acyl‐CoA dehydrogenase, and the structural basis for their different oxidative half reactions is discussed.
Soluble LDL‐R are formed by cell surface cleavage in response to phorbol esters
Authors: Begg Michael J. , Sturrock Edward D. , van der Westhuyzen Deneys R.
Published: December 2004

A 140‐kDa soluble form of the low density lipoprotein (LDL) receptor has been isolated from the culture medium of HepG2 cells and a number of other cell types. It is produced from the 160‐kDa mature LDL receptor by a proteolytic cleavage, which is stimulated in the presence of 4β‐phorbol 12‐myristate 13‐acetate (PMA), leading to the release of a soluble fragment that constitutes the bulk of the extracellular domain of the LDL receptor. By labeling HepG2 cells with [35S]methionine and chasing in the presence of PMA, we demonstrated that up to 20% of LDL‐receptors were released into the medium in a 2‐h period. Simultaneously, the level of labeled cellular receptors was reduced by 30% in those cells treated with PMA compared to untreated cells, as was the total number of cell surface LDL‐receptors assayed by the binding of 125I‐labeled antibody to whole cells. To determine if endocytosis was required for cleavage, internalization‐defective LDL‐receptors were created by mutagenesis or deletion of the NPXY internalization signal, transfected into Chinese hamster ovary cells, and assayed for cleavage in the presence and absence of PMA. Cleavage was significantly greater in the case of the mutant receptors than for wild‐type receptors, both in the absence and presence of PMA. Similar results were seen in human skin fibroblasts homozygous for each of the internalization‐defective LDL receptor phenotypes. LDL receptor cleavage was inhibited by the hydoxamate‐based inhibitor TAPI, indicating the resemblance of the LDL receptor cleavage mechanism to that of other surface released membrane proteins.
Interaction between the 2′−5′ oligoadenylate synthetase‐like protein p59 OASL and the transcriptional repressor methyl CpG‐binding protein 1
Authors: Andersen Jesper B. , Strandbygård Dorthe J. , Hartmann Rune , Justesen Just
Published: December 2004

The human 2′−5′ oligoadenylate synthetases (OAS) form a conserved family of interferon‐induced proteins consisting of four genes: OAS1, OAS2, OAS3 and the 2′−5′ oligoadenylate synthetase‐like gene (OASL). When activated by double‐stranded RNA, OAS1–3 polymerize ATP into 2′−5′‐linked oligoadenylates; 2′−5′‐linked oligoadenylates, in turn, activate a latent endoribonuclease that degrades viral and cellular RNAs. In contrast, while the p59 OASL protein is highly homologous to the OAS family (45% identity), its 350 amino acid N‐terminal domain lacks 2′−5′ oligoadenylate synthetase activity. A C‐terminal 164 amino acid domain, which is 30% homologous to a tandem repeat of ubiquitin, further distinguishes the p59 OASL protein and suggests that it serves a biological role which is distinct from other OAS family members. To dissect the function of p59 OASL, we utilized the yeast two‐hybrid system to identify interacting proteins. Methyl CpG‐binding protein 1 (MBD1), which functions as a transcriptional repressor, was identified as a strong p59 OASL interactor. Interestingly, like p59 OASL, transcription of the MBD1 gene was induced by interferon, indicating that these genes are co‐ordinately regulated. The interaction was confirmed in vitro and in vivo and was mapped to the ubiquitin‐like domain of p59 OASL. The p59 OASL–MBD1 interaction was specific, because p59 OASL did not interact with any of the other MBD family members and MBD1 did not interact with OAS1. These findings link the p59 OASL with MBD1 transcriptional control in the context of an interferon‐stimulated cell, and provide the basis for future studies to examine the functional role of this interaction.
Courtship Behavior in the Hawaiian Cricket Laupala cerasina: Males Provide Spermless Spermatophores as Nuptial Gifts
Authors: Shaw Kerry L. , Khine Albert H.
Published: December 2004

The role of sexual selection in the crickets (family Gryllidae) is well known but new forms of courtship behavior continue to be discovered. Here we describe the elaborate mating behavior of the Hawaiian endemic Laupala cerasina (subfamily Trigonidiinae) on the basis of both laboratory and field observations. We found that courtship and mating involve multiple copulations during which repeated spermatophores are transferred. Initial copulations involved the transfer of relatively small ‘microspermatophores’. The final copulation results in the transfer of a considerably larger ‘macrospermatophore’. We tested and rejected the hypothesis that the number of spermatophore transfers results in greater offspring sired by a given male. Matings involving only microspermatophores resulted in no offspring, whereas most matings involving micros and a macrospermatophore, or a macrospermatophore only, produced offspring. A direct examination of the contents of the microspermatophore revealed an absence of sperm, whereas examination of the macrospermatophore revealed the presence of sperm. Given that females consume the spermless microspermatophores, we conclude that the microspermatophores are a form of nuptial feeding of females by males. We discuss possible hypotheses to explain this unusual courtship system.
‘Defence lignin’ and hydroxycinnamyl alcohol dehydrogenase activities in wounded Eucalyptus gunnii
Authors: Hawkins S. , Boudet A.
Published: December 2004

To learn more about lignin formation in response to wounding in trees, we adopted two complementary approaches: (1) microscopic and histochemical studies of the wound response in 3.5‐month‐old Eucalyptus gunnii plantlets and (2) biochemical investigations of hydroxycinnamyl alcohol dehydrogenase activities in wounded 6‐year‐old, field‐grown E. gunnii trees. The first approach revealed that a barrier zone was formed in response to wounding in both ground tissues (cortex barrier and pith reaction zone) and vascular tissues. The barrier zone was barely detectable after 24 h but well‐developed 7 days after wounding. Microscopic analyses indicated that the barrier zone was formed by the reinforcement of cell walls with ‘lignin‐like material’ in both ground tissues and vascular tissue, and that, in addition, the lumen of certain xylem cells (vessels and fibres) were blocked by the deposition of polymeric phenolic material. Histochemical characterization revealed that the lignin‐like material (‘defence lignin’) deposited in ground tissue cell walls and xylem cell blockages was poor in syringyl (S‐type) lignin units and therefore differed from the usual mixed guaiacyl–syringyl (G–S) lignin unit composition of E. gunnii developmental lignin. In contrast, S‐type lignin appeared to be deposited in the cell walls of immature developing secondary xylem cells at a stage when the cell walls of comparable cells from unwounded control stems contained lignin poor in syringyl units. The second approach indicated that two different types of cinnamyl alcohol dehydrogenase activity are induced, and apparently regulated differentially, in response to wounding in E. gunnii trees. Coniferyl alcohol dehydrogenase activity was induced immediately and continued to increase throughout the first 15 days of the 17‐day experimental period, while sinapyl alcohol dehydrogenase activity was first detected at 8 days after wounding and continued to increase throughout the experimental period. The biological roles of the two alcohol dehydrogenase activities are discussed in relation to the formation of defence lignin versus developmental lignin in trees.
Distribution of Heterobasidion annosum intersterility groups in Poland
Authors: Łakomy P. , Werner A.
Published: December 2004

Specimens of Heterobasidion annosum were collected in 104 different stands in 43 regions of Poland. Pure cultures originating from 439 collections were identified in mating tests. Three intersterility groups, P, S and F, of H. annosum were found. Their occurrence in Poland was connected with the natural distribution of the main hosts: Pinus sylvestris, Picea abies and Abies alba, respectively. P was the most common intersterility group of H. annosum in Poland, causing mortality in Scots pine plantations and root rot in older stands. It was also isolated from Betula pendula, P. abies, Larix decidua, Fagus sylvatica and Carpinus betulus. The S group was present in the southern and north‐eastern parts of the country, causing root and butt rot mostly in spruce stands. The F group occurred in the south of Poland, in the mountains, highlands and lowland up to the northern border of the distribution of fir. It was found only on stumps, old dead trees and logs. There was no evidence of damage caused by the F group on A. alba trees.
Absence of residual effects of a defeated resistance gene in poplar
Authors: Woo K.‐S. , Newcombe G.
Published: December 2004

In a few plant pathosystems, defeated major genes have been shown to contribute to partial resistance to disease. This hypothesis has never been tested before in a forest tree, but pathogenic variation associated with recent hybridization in poplar rust in the Pacific northwest provided an opportunity. An F2 progeny of 256 poplar clones in the field near Corvallis, Oregon, USA, has been monitored for rust severity and infection type since the advent of the new hybrid rust, Melampsora × columbiana, in the mid‐1990s. All 256 clones displayed a susceptible infection type in 1997 and again in 2000, and yet variation in uredinial density (i.e. partial resistance) was still observed. To determine which clones possessed a defeated resistance gene, a greenhouse inoculation was performed with an isolate of M. medusae, one of the parents of M. × columbiana. Clones that would have been resistant to M. medusae, prior to the advent of M. × columbiana, were thus identified. The inoculation resulted in a 1 : 1 segregation (χ2=0.772; p=0.38) for resistance, indicating the presence of a major gene. However, the F2 clones possessing the defeated resistance gene displayed the same level of partial resistance in the field in both 1997 and 2000 as their full siblings lacking the gene.
Comparing Chilean and Swedish rust populations: possible impact of long‐distance transport of Melampsora rust
Authors: Hurtado S. , Ramstedt M.
Published: December 2004

As willow (Salix) cultivation grows globally, worldwide monitoring of pathotypes for Melampsora rust, the most serious disease in willow, becomes increasingly important. We compared the pathotype composition of the Melampsora rust population in Chile with that in Sweden and assessed inocula exchange for the rust between the continents. For pathotyping rust isolates, a willow differential was used (a standardized set of willow test clones) to obtain clone‐virulence patterns. These patterns were used to identify virulence components involved in determining which willow clones a given individual pathogen isolate (genotype) can successfully attack. Each virulence component detected earlier in Sweden's rust population was present in Chile's rust population. Using isoenzyme analysis to classify willow species in Chile, a low genetic variation of Salix viminalis (an introduced willow species) was found, compared with the endemic species Salix humboldtiana. Comparison of rust populations for the two countries supports the present hypothesis that intercontinental inocula exchange can be a significant determinant of local pathotype structure, and consequently can be important for willow‐resistance breeding.
Dermatologie an der Universitätsklinik Mbarara, Uganda
Authors: Schmidt Enno , Rose Christian , Mulyowa Grace K. , Jäger Gerold
Published: December 2004

Die Georg‐Klingmüller‐Klinik der Universität Mbarara wurde 1999 eröffnet. Heute werden dort jährlich circa 6 000 ambulante Patienten mit Hautkrankheiten betreut. Vor dem sozioökonomischen Hintergrund eines subsaharischen Landes soll ein Einblick gegeben werden in die Arbeit dieser im Südwesten Ugandas gelegenen Klinik. Neben dem klinischen Spektrum der Dermatosen, der Diagnostik und Therapie wird über die Bedeutung der Dermatologie für die Bekämpfung von HIV/AIDS berichtet. Es wird gezeigt, daß die Kooperation mit Hautkliniken in Deutschland sowohl die Diagnostik in Mbarara erleichtern kann als auch bei uns hilft, den Blick zu schärfen für die zunehmende Zahl von Patienten mit Erkrankungen auf dunkler Haut.
Development and retroviral transduction of porcine neonatal pancreatic islet cells in monolayer culture
Authors: Tatarkiewicz Krystyna , López‐Avalos Maria D. , Yoon Kun‐Ho , Trivedi Nitin , Quickel Robert R. , Bonner‐Weir Susan , Weir Gordon C.
Published: December 2004

To learn more about the potential of neonatal porcine pancreatic duct and islet cells for xenotransplantation, the development of these cells when cultured as monolayers was evaluated. Immunostaining for islet hormones and cytokeratin‐7 revealed that day eight monolayers consisted of approximately 70% duct cells and less than 10% beta cells. Using Ki‐67 immunostaining as a proliferation marker, the fraction of beta cells in the cell cycle was shown to decrease from 20% at day three to 10% at day eight, and for duct cells from 36 to 19%. Insulin secretion increased 2.4‐fold upon glucose stimulation, and 38‐fold when 10 mm theophylline was added, showing the responsiveness of the neonatal beta cells. Reaggregated monolayers consisted mostly of duct cells, but 4 weeks after transplantation, grafts contained predominantly endocrine cells, with duct cells being almost absent, suggesting in vivo differentiation of duct cells to endocrine cells. Monolayer susceptibility to retroviral transduction was also investigated using a Moloney Murine Leukemia Virus‐based vector. Approximately 60% of duct cells but less than 5% of beta cells expressed the transgene, indicating that precursor duct cells are better targets for transgene expression. These results show that porcine neonatal pancreatic cells can be cultured as monolayers in preparation for transplantation. Furthermore, in such a culture setting, precursor duct cells have a high rate of proliferation and are more efficiently transduced with a retrovirus‐based reporter gene than are beta cells.
Cloning and embryonic expression of six wnt genes in the medaka (Oryzias latipes) with special reference to expression of wnt5a in the pectoral fin buds
Authors: Yokoi Hayato , Nishimatsu Akiko , Ozato Kenjiro , Yoda Kinya
Published: December 2004

WNTs are secreted signaling molecules which control cell differentiation and proliferation. They are known to play essential roles in various developmental processes. Wnt genes have been identified in a variety of animals, and it has been shown that their amino acid sequences are highly conserved throughout evolution. To investigate the role of wnt genes during fish development from the evolutionary viewpoint, six medaka wnt genes (wnt4, wnt5a, wnt6, wnt7b, wnt8b and wnt8‐like) were isolated and their embryonic expression was examined. These wnt genes were expressed in various tissues during embryonic development, and most of their expression patterns were conserved or comparable to those of other vertebrates. Thus, these wnt genes may be useful as molecular markers to investigate development and organogenesis using the medaka. Focus was on wnt5a, which was expressed in the pectoral fin buds, because its expression pattern was particularly comparable to that in tetrapod limbs. Its detailed expression pattern was further examined during pectoral fin bud development. The conservation and diversification of Wnt5a expression through the evolutionary transition from fish fins to tetrapod limbs is discussed.
Putative intermediate precursor between hematogenic endothelial cells and blood cells in the developing embryo
Authors: Fraser Stuart T. , Ogawa Minetaro , Yokomizo Tomomasa , Ito Yoshiaki , Nishikawa Satomi , Nishikawa Shin‐Ichi
Published: December 2004

During embryogenesis, endothelial cells are a source of hematopoietic cells. Vascular endothelial (VE)‐cadherin modulates adherens junctions between endothelial cells. How endothelial cells, integrated into the vascular bed via adherens junctions, give rise to free‐floating hematopoietic cells has been examined. Contrary to our previous reports, in this report a cell type simultaneously expressing VE‐cadherin and the hematopoietic marker CD45 was identified, without rigorous enzymatic dissociation of embryonic tissues. In spite of expressing several other endothelial markers such as endothelial cell nitrous oxide synthase (ECNOS) and MECA‐32, this newly defined population failed to produce endothelial colonies when cultured on OP9 stroma, in direct contrast to enzymatically dissociated VE‐cadherin+ cells. When isolated from 9.5 days post coitus (d.p.c.) embryos, VE‐cadherin+ CD45+ cells generated erythroid, myeloid, but not B lymphoid, cells, also in contrast to VE‐cadherin+ cells obtained by enzymatic dissociation. Runx1 null mutant embryos lacked this novel population. Collectively, these results introduce a novel VE‐cadherin+ population within the developing embryo, which may represent an intermediate cell type in the transition of hemogenic endothelial cells into blood.
Novel pattern ofAtXloxgene expression in starfishArchaster typicusembryos
Authors: Hwang Sheng‐Ping L. , Wu Jen‐Yi , Chen Chaolun A. , Hui Cho‐Fat , Chen Chang‐Po
Published: December 2004

AnXloxhomologue gene (AtXlox) was identified in the starfishArchaster typicus. The gene consists of two exons, and encodes a polypeptide containing 228 amino acids. Although AtXlox shared 54.6 and 50.3% global amino acid sequence similarity with sea urchin SpXlox andXenopusXlHhox8, respectively, the homeodomain of AtXlox was highly conserved. Amino acid sequence identity as high as 85 to 100% was identified between the AtXlox homeodomain and its homologues from various vertebrate and invertebrate organisms. In addition, a conserved histidine residue located at position 44 of the homeodomain of all known Xlox homologues was also identified. Results of a phylogenetic analysis based on the 60 amino acid sequence of the homeodomain indicated that AtXlox was closely related to sea urchin SpXlox. Temporal developmental mRNA expression pattern analyzed by reverse transcription (RT)–polymerase chain reaction (PCR) showed thatAtXloxmRNA was mainly expressed in the early gastrula stage embryos. Whole‐mountin situhybridization revealed a ubiquitous mRNA expression pattern in archenterons as well as in ectodermal cells near the vegetal region of early and mid‐gastrula stage embryos. This spatial expression pattern is very different from those ofXloxhomologues in the leech, amphioxus, and in various vertebrate organisms with spatially restricted mRNA expression patterns in endodermal cells.
Expressional changes of ganglioside GM3 during ovarian maturation and early embryonic development in db/db mice
Authors: Kwak Dong Hoon , Jung Kyu Yong , Lee Young Choon , Choo Young Kug
Published: December 2004

Diabetes and obesity cause abnormal development of reproductive processes in a variety of species, but the mechanisms that underlie this effect have not been fully elucidated. This study examined the expressional changes of ganglioside GM3 during ovarian maturation,in vitrofertilization (IVF) and early embryonic development in diabetic/obese db/db mice. In high‐performance thin‐layer chromatography studies, GM3 expression was conspicuously low in the ovaries of db/db mice compared to non‐diabetic db/+ mice. Signal detected by anti‐GM3 monoclonal antibody was greatly reduced in the primary, secondary and graffian follicles of db/db mice compared to control mice. Results from IVF with ova and sperm from db/db mice showed that GM3 expression during early embryonic development was obviously decreased compared to db/+ mice. This study also elucidated the effects of high glucose (20 and 30 mm) on early embryonic development in ICR strain mice. High glucose caused a decrease in GM3 expression during early embryonic development. Taken together, the results of this study indicate decreased GM3 expression during ovarian maturation and embryonic development of db/db mice, suggesting that alteration of ganglioside expression induced by the diabetic condition may be implicated in the abnormal follicular embryonic development.
Is endoscopic one‐piece mucosal resection essential for early gastric cancer?
Authors: EGUCHI TAKAKO , GOTODA TAKUJI , ODA ICHIRO , HAMANAKA HISANAO , HASUIKE NORIAKI , SAITO DAIZO
Published: December 2004

Background:  Endoscopic mucosal resection (EMR) is widely accepted as a minimally invasive treatment for early gastric cancer (EGC) in Japan. However, the criteria for EMR must be strictly adhered to otherwise patients will miss the chance for additional therapy. We assess the important factor in expanding the indication of EMR.

Methods:  We investigated 1101 EGCs that had been resected by EMR at the National Cancer Center Hospital (NCCH), Tokyo, Japan, according to the indication recommended by Japanese Gastric Cancer Association (JGCA) and the expanded indication proposed by NCCH. Curability and local recurrence of the EMRs were assessed related to the applied indication and the number of resected specimens.

Results:  The recurrence rate of non‐evaluable resection was higher than that of evaluable resection (P < 0.0001). Eighty‐three lesions among 772 lesions in the JGCA group were non‐evaluable. Thirty‐seven leisons among 329 lesions in the NCCH group were non‐evaluable. There was no difference in the rate of non‐evaluable resection between JGCA and NCCH groups (P = 0.8329). However, the rate of curative resection was lower in the NCCH group than in the JGCA group (P = 0.0009). In piecemeal resection, there was no difference in the rate of non‐evaluable resection between JGCA and NCCH groups (P = 0.0527). In one‐piece resection, the rate of non‐evaluable resection was lower in the NCCH group than the JGCA group (P = 0.0137).

Conclusion:  Based on our series of cases, we propose one‐piece resection as a gold standard for EMR because it enables accurate histological evaluation, even in the EMR, according to the expanded indication.
Analysis of urgent colonoscopy for acute lower intestinal bleeding
Authors: FUJISHIRO HIROFUMI , ADACHI KYOICHI , IMAOKA TOMONORI , KOHGE NARUAKI , KAWAMURA AKIRA , KOMAZAWA YOSHINORI , ONO MASAHIRO , YUKI MIKA , SATO HIROSHI , AMANO YUJI , ISHIHARA SHUNJI , KINOSHITA YOSHIKAZU
Published: December 2004

Background:  Urgent colonoscopy is often performed to diagnose and stop acute lower intestinal bleeding, but its usefulness has yet to be clarified. The aim of this study was to clarify the present role of urgent colonoscopy for this purpose.

Methods  Urgent colonoscopy was defined as colonoscopy performed within 48 h of the occurrence of hematochezia. All patients investigated by urgent colonoscopy from September 1996 to March 2000 who were hospitalized in Shimane Medical University Hospital or Shimane Prefectural Central Hospital, Japan, were enrolled in this study. The endoscopic data and medical records of all the investigated patients were reviewed retrospectively to clarify the role of urgent colonoscopy for diagnosing and treating patients with hematochezia.

Results  A total of 152 patients were enrolled and 137 (90.1%) were successfully diagnosed endoscopically. Ischemic colitis was the most frequent disease found (32.9%), followed by postpolypectomy hemorrhage (9.2%), hemorrhoids (8.6%) and rectal ulcer (7.2%). Endoscopic hemostasis was employed to treat postpolypectomy hemorrhage and bleeding rectal ulcers with favorable results.

Conclusions  Urgent colonoscopy is useful for the diagnosis of hematochezia. Endoscopic hemostasis is frequently required to treat postpolypectomy hemorrhage and bleeding rectal ulcers.
Epidemic enterocolitis possibly as a result of Norwalk virus infection presenting as ischemic colitis
Authors: MATSUHASHI NOBUYUKI , NISHI YUMIKO , NAGOSHI DAIKI , KANEMOTO HIDEYUKI , NAKAJIMA ATSUSHI
Published: December 2004

Ischemic colitis usually presents as an isolated event, and clustering seldom occurs. Although a majority of instances of colonic ischemia have no clearly identifiable cause, many kinds of predisposing factors for the occurrence of ischemic colitis are pointed out. These include high age, generalized atherosclerosis, impaired cardiac function, hypovolemia, hypotension, colonoscopy, obstructing colonic cancer, coagulopathies and use of vasoconstrictors or contraceptives. However, infectious viral enterocolitis has not been regarded as a predisposing factor for colonic ischemia. We present here two cases of epidemic enterocolitis presenting as ischemic colitis that occurred almost simultaneously in two compromized long‐term patients in Noda Hospital, Chiba Prefecture. At the time, viral enterocolitis due to Norwalk virus infection was epidemic in Japan, including in the region of the hospital. It is likely that epidemic enterocolitis induced strong gut peristalsis and caused colonic spasm that mediated colonic ischemia. Indeed, both patients were elderly and had prominent signs of atherosclerosis, and one patient suffered from diabetes mellitus. Notably, the physician who performed the rectosigmoidoscopy in the two patients presented with enterocolitis 1 day after the procedure. This indicates that occupational transmission of epidemic enterocolitis can occur in medical staff during endoscopy procedures.
Identification of quorum‐sensing regulated proteins in the opportunistic pathogen Pseudomonas aeruginosa by proteomics
Authors: Arevalo‐Ferro Catalina , Hentzer Morten , Reil Gerold , Görg Angelika , Kjelleberg Staffan , Givskov Michael , Riedel Kathrin , Eberl Leo
Published: December 2004

The Gram‐negative bacterium Pseudomonas aeruginosa is an opportunistic human pathogen which is responsible for severe nosocomial infections in immunocompromised patients and is the major pathogen in cystic fibrosis. The bacterium utilizes two interrelated quorum‐sensing (QS) systems, which rely on N‐acyl‐homoserine lactone (AHL) signal molecules, to control the expression of virulence factors and biofilm development. In this study, we compared the protein patterns of the intracellular, extracellular and surface protein fractions of the PAO1 parent strain with those of an isogenic lasI rhlI double mutant by means of two‐dimensional gel electrophoresis (2‐DE). This analysis showed that the intensities of 23.7% of all detected protein spots differed more than 2.5‐fold between the two strains. We only considered those protein spots truly QS regulated that were changed in the mutant in the absence of signal molecules but were rescued to the wild‐type situation when the medium was supplemented with AHLs. These protein spots were characterized by MALDI‐TOF peptide mapping. Twenty‐seven proteins were identified that were previously reported to be AHL controlled, among them several well‐characterized virulence factors. For one of the identified proteins, the serine protease PrpL, a biochemical assay was established to verify that expression of this factor is indeed QS regulated. Furthermore, it is shown that the quorum‐sensing blocker C‐30 specifically interferes with the expression of 67% of the AHL‐controlled protein spots of the surface fraction, confirming the high specificity of the compound. Importantly, 20 novel QS‐regulated proteins were identified, many of which are involved in iron utilization, suggesting a link between quorum sensing and the iron regulatory system. Two of these proteins, PhuR and HasAp, are components of the two distinct haem‐uptake systems present in P. aeruginosa. In agreement with the finding that both proteins are positively regulated by the QS cascade, we show that the lasI rhlI double mutant grows poorly with haemoglobin as the only iron source when compared with the wild type. These results add haemoglobin utilization to the list of phenotypes controlled through QS in P. aeruginosa. The surprisingly high number of AHL‐regulated proteins relative to the number of regulated genes suggests that quorum‐sensing control also operates via post‐transcriptional mechanisms. To strengthen this hypothesis we investigated the role of quorum sensing in the post‐translational modification of HasAp, an extracellular protein required for the uptake of free and haemoglobin‐bound haem.
Genetic diversity and population structure of Aeromonas hydrophila, Aer. bestiarum, Aer. salmonicida and Aer. popoffii by multilocus enzyme electrophoresis (MLEE)
Authors: Miñana‐Galbis David , Farfán Maribel , Fusté M. Carme , Lorén J. Gaspar
Published: December 2004

Genetic diversity, genetic relationship, identification and population structure of 120 Aeromonas strains (including Aer. hydrophila, Aer. bestiarum, Aer. salmonicida and Aer. popoffii) isolated from various sources were studied by analysis of 15 genetic loci by multilocus enzyme electrophoresis (MLEE). All 15 loci were polymorphic, with an average of 9.4 alleles per locus and a mean genetic diversity (H) of 0.64. Cluster analysis defined at H ≤ 0.7 differentiated most of the taxa analysed except the Aer. popoffii and Aer. bestiarum strains, which showed a close genetic relationship. Allelic frequencies of five loci (EST1, HEX, IDH, LDH1 and MDH) identified 94% of the strains. The index of association (IA) for the total sample was 2.38 and IA values calculated for the different populations were always significantly different from zero. These results suggest that the population structure of this Aeromonas sample is strongly clonal, confirm the taxonomic status of the analysed species in population genetics terms, and show the usefulness of MLEE for identifying Aeromonas species.
The impact of diabetes and obesity on liver histology in patients with hepatitis C
Authors: Friedenberg F. , Pungpapong S. , Zaeri N. , Braitman L. E.
Published: December 2004

Aim:  An association between diabetes mellitus and HCV has been recognized previously. No study has examined whether there is an independent association between the degree of hepatic fibrosis and the incidence of diabetes in HCV patients when controlling for other risk factors.

Methods:  We reviewed the charts of 264 consecutive patients with chronic HCV infection at a referral liver centre from January 1991 to December 1999. Demographic background, medical history, laboratory and liver biopsy results were retrieved.

Results:  The prevalence of diabetes was 16.3%. Gender, intravenous drug use, steatosis scores, aminotransferase levels and iron studies were similarly distributed in patients with and without diabetes (all p > 0.05). In contrast, mean age was greater in the diabetic group (49.8 vs. 44.3, p = 0.003). The prevalence of diabetes was substantially higher in African‐Americans (p = 0.001) and those with BMI > 30 (p = 0.015). Although the fibrosis score was higher in diabetics (ρ = 0.14, p = 0.03), that association did not remain significant when controlling for diabetes risk factors (p > 0.3). The degree of steatosis and fibrosis both tended to increase with increasing BMI (ρ = 0.47, p < 0.001 and ρ = 0.13, p = 0.03, respectively). Even after controlling for diabetes, age, gender, race, and current alcohol use, those associations remained (both p < 0.001).

Conclusions:  The prevalence of diabetes in our group of HCV patients was high, consistent with other studies. Diabetes is not an independent predictor of degree of fibrosis. Body mass index is an independent predictor of both fibrosis and steatosis in HCV patients.
Comparison of intraperitoneal and posterior subcutaneous abdominal adipose tissue compartments as predictors of VLDL apolipoprotein B‐100 kinetics in overweight/obese men
Authors: Chan D. C. , Watts G. F. , Barrett P. H. R.
Published: December 2004

Aims:  The influence of different regional adipose tissue (AT) compartments on insulin resistance and dyslipidaemia may account for variations in risk of diabetes and cardiovascular disease. The purpose of this study was to examine the association of intraperitoneal and posterior subcutaneous abdominal AT with very‐low‐density lipoprotein apolipoprotein B‐100 (VLDL‐apoB) kinetics in overweight/obese men.

Methods:  Intraperitoneal, anterior subcutaneous and posterior subcutaneous abdominal AT mass (IPATM, ASAATM and PSAATM respectively) were quantified in 51 overweight/obese men using magnetic resonance imaging. Hepatic secretion of VLDL‐apoB was measured using an intravenous infusion of 1‐[13C]‐leucine. Isotopic enrichment of VLDL‐apoB was measured using gas chromatography mass spectrometry and a multicompartmental model used to estimate VLDL‐apoB metabolic parameters. Insulin resistance was estimated by homeostasis model assessment (HOMA) score.

Results:  In univariate analysis, IPATM, ASAATM and PSAATM were significantly associated with HOMA score (r = 0.554, 0.425 and 0.440 respectively; p < 0.01). Intraperitoneal abdominal AT mass was also associated with plasma triglycerides (r = 0.292, p < 0.05), VLDL‐apoB concentrations (r = 0.407, p < 0.01) and VLDL‐apoB secretion (r = 0.390, p < 0.05). Intraperitoneal abdominal AT mass remained significantly associated with VLDL‐apoB secretion (r = 0.344, p < 0.05) and HOMA score (r = 0.368, p < 0.01) after adjusting for total body fat. In multiple regression analysis including IPATM, non‐esterified fatty acids and age, IPATM was the best predictor of VLDL‐apoB secretion (R2 = 17%, p < 0.01) and HOMA score (R2 = 32%, p < 0.001). None of the fat compartments were significantly associated with VLDL‐apoB catabolism.

Conclusions:  In overweight/obese men the intraperitoneal AT mass is a better predictor of VLDL‐apoB secretion and insulin resistance than either posterior or anterior subcutaneous abdominal AT mass.
Metformin inhibits glycogen synthesis and gluconeogenesis in cultured rat hepatocytes
Authors: Otto M. , Breinholt J. , Westergaard N.
Published: December 2004

Aim:  Glycogen synthesis, and glucose and lactate production were examined in cultured rat hepatocytes preincubated with metformin (0–500 µm) for 24 h.

Methods:  Cells incubated with[1‐13C]‐glucose and [1‐13C]‐lactate allowed us to study the effect of metformin on glucose production from glycogenolysis and gluconeogenesis in a detailed manner using NMR spectroscopy. 1H and 13C‐filtered 1H‐NMR spectra were recorded by using flow‐injection technique.

Results:  Metformin decreased glycogen synthesis in a dose‐dependent manner with an IC50 value of 196.5 µm. This effect could not be reversed by the presence of the glycogen phosphorylase inhibitor DAB, suggesting that glycogenolysis was not affected. A clear correlation between glucose production and glycogen content (0.97 < R < 0.99; p < 0.001) and lactate production and glycogen content (0.97 < R < 0.99; p < 0.001) was observed. Moreover, a strong inhibition (62%, p < 0.001) of glucose produced from lactate/pyruvate (3 mm/0.3 mm) was observed in cells treated with 350 µm metformin.

Conclusion:  Hepatocytes preincubated for 24 h in the presence of metformin at clinically relevant concentrations showed impaired glycogenesis as well as gluconeogenesis.