bluuebunny/crossref_metadata_2025 huggingface dataset:
Transcriptional regulation of the human TRIF (TIR domain-containing adaptor protein inducing interferon beta) gene
Authors: HARDY Matthew P. , McGETTRICK Anne F. , O'NEILL Luke A. J.
Published: June 2004

TRIF [TIR (Toll/interleukin-1 receptor) domain-containing adaptor protein inducing interferon β; also known as TICAM-1 (TIR-containing adaptor molecule-1)] is a key adaptor for TLR3 (Toll-like receptor 3)- and TLR4-mediated signalling. We have performed a detailed annotation of the human TRIF gene and fine analysis of the basal and inducible promoter elements lying 5´ to the site of initiation of transcription. Human TRIF maps to chromosome 19p13.3 and is flanked upstream by TIP47, which encodes the mannose 6-phosphate receptor binding protein, and downstream by a gene encoding FEM1a, a human homologue of the Caenorhabditis elegans Feminisation-1 gene. Using promoter–reporter deletion constructs, we identified a distal region with the ability to negatively regulate basal transcription and a proximal region containing an Sp1 (stimulating protein 1) site that confers approx. 75% of basal transcriptional activity. TRIF expression can be induced by multiple stimuli, such as the ligands for TLR2, TLR3 and TLR4, and by the pro-inflammatory cytokines tumour necrosis factor α and interleukin-1α. All of these stimuli act via an NF-κB (nuclear factor-κB) motif at position −127. In spite of the presence of a STAT1 (signal transduction and activators of transcription 1) motif at position −330, the addition of type I or type II interferon had no effect on TRIF activity. The human TRIF gene would therefore appear to be regulated primarily by NF-κB.
Biosynthesis of anandamide and N-palmitoylethanolamine by sequential actions of phospholipase A2 and lysophospholipase D
Authors: SUN Yong-Xin , TSUBOI Kazuhito , OKAMOTO Yasuo , TONAI Takeharu , MURAKAMI Makoto , KUDO Ichiro , UEDA Natsuo
Published: June 2004

Anandamide (an endocannabinoid) and other bioactive long-chain NAEs (N-acylethanolamines) are formed by direct release from N-acyl-PE (N-acyl-phosphatidylethanolamine) by a PLD (phospholipase D). However, the possible presence of a two-step pathway from N-acyl-PE has also been suggested previously, which comprises (1) the hydrolysis of N-acyl-PE to N-acyl-lysoPE by PLA1/PLA2 enzyme(s) and (2) the release of NAEs from N-acyllysoPE by lysoPLD (lysophospholipase D) enzyme(s). In the present study we report for the first time the characterization of enzymes responsible for this pathway. The PLA1/PLA2 activity for N-palmitoyl-PE was found in various rat tissues, with the highest activity in the stomach. This stomach enzyme was identified as group IB sPLA2 (secretory PLA2), and its product was determined as N-acyl-1-acyl-lysoPE. Recombinant group IB, IIA and V of sPLA2s were also active with N-palmitoyl-PE, whereas group X sPLA2 and cytosolic PLA2α were inactive. In addition, we found wide distribution of lysoPLD activity generating N-palmitoylethanolamine from N-palmitoyl-lysoPE in rat tissues, with higher activities in the brain and testis. Based on several lines of enzymological evidence, the lysoPLD enzyme could be distinct from the known N-acyl-PE-hydrolysing PLD. sPLA2-IB dose dependently enhanced the production of N-palmitoylethanolamine from N-palmitoyl-PE in the brain homogenate showing the lysoPLD activity. N-Arachidonoyl-PE and N-arachidonoyl-lysoPE as anandamide precursors were also good substrates of sPLA2-IB and the lysoPLD respectively. These results suggest that the sequential actions of PLA2 and lysoPLD may constitute another biosynthetic pathway for NAEs, including anandamide.
Expression pattern of the deoxyribonuclease 1 gene: lessons from the Dnase1 knockout mouse
Authors: NAPIREI Markus , RICKEN Albert , EULITZ Dirk , KNOOP Heiko , MANNHERZ Hans Georg
Published: June 2004

The tissue distribution of deoxyribonuclease 1 (DNASE1, DNase I), a Ca2+ and Mg2+/Mn2+-dependent secretory endonuclease, has previously been investigated. However, most of these studies did not account for the existence of different members of the DNASE1 gene family, did not differentiate between endogenous DNASE1 protein synthesis and its extracellular occurrence or were not performed with methods allowing both a sensitive and a specific detection. Now we re-examined the DNASE1 gene expression pattern by taking advantage of the Dnase1 knockout mouse model. Direct comparison of samples derived from wild-type (Dnase1+/+) and knockout (Dnase1−/−) mice allowed an unambiguous detection of Dnase1 gene expression at the mRNA and protein level. For the detection of Dnase1 activity, we developed a highly sensitive nuclease zymogram method. We observed high Dnase1 gene expression in the parotid and submandibular gland as well as in the kidney and duodenum, intermediate expression in the ileum, mesenterial lymph nodes, liver, ventral prostate, epididymis, ovary and stomach, and low expression in the sublingual, preputial, coagulation and pituitary gland. We report for the first time the lachrymal and thyroid glands, the urinary bladder and the eye to be Dnase1-expressing organs as well. Since Dnase1 knockout mice with the 129xC57Bl/6 mixed genetic background have indicated the protection against an anti-DNA autoimmune response as a new physiological function of Dnase1, knowledge of the physiological sites of its synthesis might prove helpful to find new therapeutic strategies.
Epigenetic modifications affect Dnmt3L expression
Authors: AAPOLA Ulla , MÄENPÄÄ Katja , KAIPIA Antti , PETERSON Pärt
Published: June 2004

Imprinted genes are expressed from a single allele due to differential methylation of maternal or paternal alleles during gametogenesis. Dnmt3L (DNA cytosine-5-methyltransferase 3 like), a member of de novo methyltransferase Dnmt3 protein family, is a regulator of maternal imprinting. In the present study, we have characterized the promoter region of the mouse Dnmt3L gene. Transient transfection assays performed with 5´-deletion promoter constructs indicated a minimal promoter area within 440 bp upstream from the translational start site. Longer promoter constructs showed decreased activity, suggesting the presence of repressor elements within the upstream sequences. According to electrophoretic mobility-shift assays and mutation analysis, the minimal promoter region contained four functional binding sites for the Sp1 (specificity protein 1) family of transcription factors, Sp1 and Sp3. In vitro methylation of Dnmt3L promoter constructs decreased the transcriptional activity significantly, demonstrating down-regulation by cytosine methylation. This was supported by the results from bisulphite sequencing and real-time quantitative reverse transcriptase–PCR analysis of different mouse cell lines and tissues. In testis and embryonic stem cells showing strong Dnmt3L expression, all CpG sites studied were fully unmethylated, whereas non-expressive cell lines and tissues with lesser Dnmt3L expression showed complete or diverse CpG methylation levels. Treatment of Dnmt3L non-expressive cell lines with deacetylase inhibitor trichostatin A and methyltransferase inhibitor 5-aza-2´-deoxycytidine induced the expression of Dnmt3L mRNA. Furthermore, we show that the repressional effect of longer promoter fragments was also relieved by these inhibitors, altogether indicating an epigenetic control for Dnmt3L gene regulation.
Lysophosphatidylcholine and 7-oxocholesterol modulate Ca2+ signals and inhibit the phosphorylation of endothelial NO synthase and cytosolic phospholipase A2
Authors: MILLANVOYE-VAN BRUSSEL Elisabeth , TOPAL Gökce , BRUNET Annie , PHAM Thuc DO , DECKERT Valérie , RENDU Francine , DAVID-DUFILHO Monique
Published: June 2004

The oxidation of plasma LDLs (low-density lipoproteins) is a key event in the pathogenesis of atherosclerosis. LPC (lysophosphatidylcholine) and oxysterols are major lipid constitutents of oxidized LDLs. In particular, 7-oxocholesterol has been found in plasma from cardiac patients and atherosclerotic plaque. In the present study, we investigated the ability of 7-oxocholesterol and LPC to regulate the activation of eNOS (endothelial nitric oxide synthase) and cPLA2 (cytosolic phospholipase A2) that synthesize two essential factors for vascular wall integrity, NO (nitric oxide) and arachidonic acid. In endothelial cells from human umbilical vein cords, both 7-oxocholesterol (150 µM) and LPC (20 µM) decreased histamine-induced NO release, but not the release activated by thapsigargin. The two lipids decreased NO release through a PI3K (phosphoinositide 3-kinase)-dependent pathway, and decreased eNOS phosphorylation. Their mechanisms of action were, however, different. The NO release reduction was dependent on superoxide anions in LPC-treated cells and not in 7-oxocholesterol-treated ones. The Ca2+ signals induced by histamine were abolished by LPC, but not by 7-oxocholesterol. The oxysterol also inhibited (i) the histamine- and thapsigargin-induced arachidonic acid release, and (ii) the phosphorylation of both cPLA2 and ERK1/2 (extracellular-signal-regulated kinases 1/2). The results show that 7-oxocholesterol inhibits eNOS and cPLA2 activation by altering a Ca2+-independent upstream step of PI3K and ERK1/2 cascades, whereas LPC desensitizes eNOS by interfering with receptor-activated signalling pathways. This suggests that 7-oxocholesterol and LPC generate signals which cross-talk with heterologous receptors, effects which could appear at early stage of atherosclerosis.
Antibody probe study of Ca2+ channel regulation by interdomain interaction within the ryanodine receptor
Authors: KOBAYASHI Shigeki , YAMAMOTO Takeshi , PARNESS Jerome , IKEMOTO Noriaki
Published: June 2004

N-terminal and central domains of ryanodine receptor 1 (RyR1), where many reported malignant hyperthermia (MH) mutations are localized, represent putative channel regulatory domains. Recent domain peptide (DP) probe studies led us to the hypothesis that these domains interact to stabilize the closed state of channel (zipping), while weakening of domain–domain interactions (unzipping) by mutation de-stabilizes the channel, making it leaky to Ca2+ or sensitive to the agonists of RyR1. As shown previously, DP1 (N-terminal domain peptide) and DP4 (central domain peptide) produced MH-like channel activation/sensitization effects, presumably by peptide binding to sites critical to stabilizing domain–domain interactions and resultant loss of conformational constraints. Here we report that polyclonal anti-DP1 and anti-DP4 antibodies also produce MH-like channel activation and sensitization effects as evidenced by about 4-fold enhancement of high affinity [3H]ryanodine binding to RyR1 and by a significant left-shift of the concentration-dependence of activation of sarcoplasmic reticulum Ca2+ release by polylysine. Fluorescence quenching experiments demonstrate that the accessibility of a DP4-directed, conformationally sensitive fluorescence probe linked to the RyR1 N-terminal domain is increased in the presence of domain-specific antibodies, consistent with the view that these antibodies produce unzipping of interacting domains that are of hindered accessibility to the surrounding aqueous environment. Our results suggest that domain-specific antibody binding induces a conformational change resulting in channel activation, and are consistent with the hypothesis that interacting N-terminal and central domains are intimately involved in the regulation of RyR1 channel function.
In vitro synthesis of a crystalline (1 3,1 4)-beta-d-glucan by a mutated (1 3,1 4)-beta-d-glucanase from Bacillus
Authors: FAIJES Magda , IMAI Tomoya , BULONE Vincent , PLANAS Antoni
Published: June 2004

Oligo- and poly-saccharides have a large number of important biological functions, and they occur in natural composite materials, such as plant cell walls, where they self-assemble during biosynthesis in a poorly understood manner. They can also be used for the formation of artificial composite materials with industrial applications. Fundamental and applied research in biology and nanobiotechnology would benefit from the possibility of synthesizing tailor-made oligo-/poly-saccharides. In the present paper, we demonstrate that such syntheses are possible using genetically modified glycoside hydrolases, i.e. glycosynthases. The ability of the endoglycosynthase derived from Bacillus (1→3,1→4)-β-d-glucanase to catalyse self-condensation of sugar donors was exploited for the in vitro synthesis of a regular polysaccharide. The specificity of the enzyme allowed the polymerization of α-laminaribiosyl fluoride via the formation of (1→4)-β-linkages to yield a new linear crystalline (1→3,1→4)-β-d-glucan with a repeating 4βG3βG unit. MS and methylation analyses indicated that the in vitro product consisted of a mixture of oligosaccharides, the one having a degree of polymerization of 12 being the most abundant. Morphological characterization revealed that the (1→3,1→4)-β-d-glucan forms spherulites which are composed of platelet crystals. X-ray and electron diffraction analyses allowed the proposition of a putative crystallographic structure which corresponds to a monoclinic unit cell with a=0.834 nm, b=0.825 nm, c=2.04 nm and γ=90.5°. The dimensions of the ab plane are similar to those of cellulose Iβ, but the length of the c-axis is nearly twice that of cellulose I. It is proposed that four glucose residues are present in an extended conformation along the c-axis of the unit cell. The data presented show that glycosynthases represent promising enzymic systems for the synthesis of novel polysaccharides with specific and controlled structures, and for the analysis in vitro of the mechanisms of polymerization and crystallization of polysaccharides.
Actin depolymerization-induced tyrosine phosphorylation of cortactin: the role of Fer kinase
Authors: FAN Lingzhi , Di CIANO-OLIVEIRA Caterina , WEED Scott A. , CRAIG Andrew W. B. , GREER Peter A. , ROTSTEIN Ori D. , KAPUS András
Published: June 2004

The F-actin-binding protein cortactin is an important regulator of cytoskeletal dynamics, and a prominent target of various tyrosine kinases. Tyrosine phosphorylation of cortactin has been suggested to reduce its F-actin cross-linking capability. In the present study, we investigated whether a reciprocal relationship exists, i.e. whether the polymerization state of actin impacts on the cortactin tyrosine phosphorylation. Actin depolymerization by LB (latrunculin B) induced robust phosphorylation of C-terminal tyrosine residues of cortactin. In contrast, F-actin stabilization by jasplakinolide, which redistributed cortactin to F-actin-containing patches, prevented cortactin phosphorylation triggered by hypertonic stress or LB. Using cell lines deficient in candidate tyrosine kinases, we found that the F-actin depolymerization-induced cortactin phosphorylation was mediated by the Fyn/Fer kinase pathway, independent of Src and c-Abl. LB caused modest Fer activation and strongly facilitated the association between Fer and cortactin. Interestingly, the F-actin-binding region within the cortactin N-terminus was essential for the efficient phosphorylation of C-terminal tyrosine residues. Investigating the structural requirements for the Fer–cortactin association, we found that (i) phosphorylation-incompetent cortactin still bound to Fer; (ii) the isolated N-terminus associated with Fer; and (iii) the C-terminus alone was insufficient for binding. Thus the cortactin N-terminus participates in the Fer–cortactin interaction, which cannot be fully due to the binding of the Fer Src homology 2 domain to C-terminal tyrosine residues of cortactin. Taken together, F-actin stabilization prevents cortactin tyrosine phosphorylation, whereas depolymerization promotes it. Depolymerization-induced phosphorylation is mediated by Fer, and requires the actin-binding domain of cortactin. These results define a novel F-actin-dependent pathway that may serve as a feedback mechanism during cytoskeleton remodelling.
A functional polymeric immunoglobulin receptor in chicken (Gallus gallus) indicates ancient role of secretory IgA in mucosal immunity
Authors: WIELAND Willemien H. , ORZÁEZ Diego , LAMMERS Aart , PARMENTIER Henk K. , VERSTEGEN Martin W. A. , SCHOTS Arjen
Published: June 2004

Animals are continuously threatened by pathogens entering the body through natural openings. Here we show that in chicken (Gallus gallus), secretory IgA (sIgA) protects the epithelia lining these natural cavities. A gene encoding a chicken polymeric Ig receptor (GG-pIgR), a key component of sIgA, was identified, and shown to be expressed in the liver, intestine and bursa of Fabricius. All motifs involved in pIgR function are present, with a highly conserved Ig-binding motif in the first Ig-like domain. Physical association of GG-pIgR with pIgA in bile and intestine demonstrates that this protein is a functional receptor. Thus, as shown for mammals, this receptor interacts with J-chain-containing polymeric IgA (pIgA) at the basolateral epithelial cell surface resulting in transcytosis and subsequent cleavage of the pIgR, releasing sIgA in the mucosal lumen. Interestingly, the extracellular portion of GG-pIgR protein comprises only four Ig-like domains, in contrast with the five domain structure found in mammalian pIgR genes. The second Ig-like domain of mammalian pIgR does not have an orthologous domain in the chicken gene. The presence of pIgR in chicken suggests that this gene has evolved before the divergence of birds and reptiles, indicating that secretory Igs may have a prominent role in first line defence in various non-mammalian species.
Identification of a stretch of six divergent amino acids on the alpha5 helix of Galpha16 as a major determinant of the promiscuity and efficiency of receptor coupling
Authors: HO Maurice K. C. , CHAN Jasmine H. P. , WONG Cecilia S. S. , WONG Yung H.
Published: June 2004

A broad repertory of G-protein-coupled receptors shows effective coupling with the haematopoietic G16 protein. In the present study, individual residues along the C-terminal α5 helix of Gα16 were examined for their contributions in defining receptor-coupling specificity. Residues that are relatively conserved within, but diverse between, the subfamilies of cloned Gα subunits were mutated into the corresponding Gαz residues. Six Gi-linked receptors with different coupling efficiencies to Gα16 were examined for their ability to utilize the various Gα16 mutants to mediate agonist-induced inositol phosphate accumulation and Ca2+ mobilization. Co-operative enhancements of receptor coupling were observed with chimaeras harbouring multiple mutations at Glu350, Lys357 and Leu364 of Gα16. Mutation of Leu364 into isoleucine appeared to be more efficient in enhancing receptor recognition compared with mutations at the other two sites. Mutation of a stretch of six consecutive residues (362–367) lying towards the end of the α5 helix was found to broaden significantly the receptor-coupling profile of Gα16, and the effect was mediated partly through interactions with the β2–β3 loop. These results suggested that a stretch of six distinctive residues at the α5 helix of Gα16 is particularly important, whereas other discrete residues spreading along the α5 helix function co-operatively for determining the specificity of receptor recognition.
Parasite-specific eIF2 (eukaryotic initiation factor-2) kinase required for stress-induced translation control
Authors: SULLIVAN William J. , NARASIMHAN Jana , BHATTI Micah M. , WEK Ronald C.
Published: June 2004

The ubiquitous intracellular parasite Toxoplasma gondii (phylum Apicomplexa) differentiates into an encysted form (bradyzoite) that can repeatedly re-emerge as a life-threatening acute infection (tachyzoite) upon impairment of immunity. Since the switch from tachyzoite to bradyzoite is a stress-induced response, we sought to identify components related to the phosphorylation of the α subunit of eIF2 (eukaryotic initiation factor-2), a well-characterized event associated with stress remediation in other eukaryotic systems. In addition to characterizing Toxoplasma eIF2α (TgIF2α), we have discovered a novel eIF2 protein kinase, designated TgIF2K-A (Toxoplasma gondiiinitiation factor-2kinase). Although the catalytic domain of TgIF2K-A contains sequence and structural features that are conserved among members of the eIF2 kinase family, TgIF2K-A has an extended N-terminal region that is highly divergent from other eIF2 kinases. TgIF2K-A specifically phosphorylates the regulatory serine residue of yeast eIF2α in vitro and in vivo, and can modulate translation when expressed in the yeast model system. We also demonstrate that TgIF2K-A phosphorylates the analogous regulatory serine residue of recombinant TgIF2α in vitro. Finally, we demonstrate that TgIF2α phosphorylation in tachyzoites is enhanced in response to heat shock or alkaline stress, conditions known to induce parasite differentiation in vitro. Collectively, this study suggests that eIF2 kinase-mediated stress responses are conserved in Apicomplexa, and a novel family member exists that may control parasite-specific events, including the clinically relevant conversion into bradyzoite cysts.
Truncated variants of hyaluronan-binding protein 1 bind hyaluronan and induce identical morphological aberrations in COS-1 cells
Authors: SENGUPTA Aniruddha , TYAGI Rakesh K. , DATTA Kasturi
Published: June 2004

Hyaluronan (HA)-binding protein 1 (HABP1) is multifunctional in nature and exists as a trimer through coiled-coil interaction between α-helices at its N- and C-termini. To investigate the importance of trimeric assemblage and HA-binding ability of HABP1, we generated and overexpressed variants of HABP1 by truncating the α-helices at its termini. Subsequently, these variants were transiently expressed in COS-1 cells to examine the influence of these structural variations on normal cell morphology, as compared with those imparted by HABP1. Substantiating the centrality of coiled-coil interaction for maintaining the trimeric assembly of HABP1, we demonstrate that disruption of trimerization does not alter the affinity of variants towards its ligand HA. Transient expression of HABP1 altered the morphology of COS-1 cells by generating numerous cytoplasmic vacuoles along with disruption of the f-actin network. Interestingly, the truncated variants also imparted identical morphological changes. Characterization of the cytoplasmic vacuoles revealed that most of these vacuoles were autophagic in nature, resembling those generated under stress conditions. The identical morphological changes manifested in COS-1 cells on transient expression of HABP1 or its variants is attributed to their comparable HA-binding ability, which in concert with endogenous HABP1, may deplete the cellular HA pool. Such quenching of HA below a threshold level in the cellular milieu could generate a stress condition, manifested through cytoplasmic vacuoles and a disassembly of the f-actin network.
Alternative reading frame protein (ARF)-independent function of CARF (collaborator of ARF) involves its interactions with p53: evidence for a novel p53-activation pathway and its negative feedback control
Authors: HASAN M. Kamrul , YAGUCHI Tomoko , MINODA Yasumasu , HIRANO Takashi , TAIRA Kazunari , WADHWA Renu , KAUL Sunil C.
Published: June 2004

CARF, a collaborator of ARF (alternative reading frame protein), was cloned as a novel ARF-binding protein from a yeast-interaction screen. It potentiated ARF-mediated p53 function, and also caused a moderate increase in p53 activity in the absence of ARF. We herein report the molecular mechanism of ARF-independent function of CARF. By employing a variety of approaches, including overexpression of CARF, its suppression by small interfering RNA and use of protease inhibitors, we demonstrate that: (i) CARF directly interacts with wild-type p53, causing its stabilization and functional activation; and (ii) CARF and p53 levels show an inverse relationship that is instigated by a negative-feedback control via a proteasome-mediated degradation pathway.
Unravelling calcium-release channel gating: clues from a hot disease
Authors: McCARTHY Tommie V. , MACKRILL John J.
Published: June 2004

Ryanodine receptors (RyRs) are a family of intracellular channels that mediate Ca2+ release from the endoplasmic and sarcoplasmic reticulum. More than 50 distinct point mutations in one member of this family, RyR1, cause malignant hyperthermia, a potentially lethal pharmacogenetic disorder of skeletal muscle. These mutations are not randomly distributed throughout the primary structure of RyR1, but are grouped in three discrete clusters. In this issue of the Biochemical Journal, Kobayashi et al. present evidence that interdomain interactions between two of these mutation-enriched regions play a key role in the gating mechanism of RyR1.
Insulin signalling: putting the G- in protein-protein interactions
Authors: MALBON Craig C.
Published: June 2004

Cell signalling via receptor tyrosine kinases, such as the insulin receptor, and via heterotrimeric G-proteins, such as Gαi, Gαs and Gαq family members, constitute two of most avidly studied paradigms in cell biology. That elements of these two populous signalling pathways must cross-talk to achieve proper signalling in the regulation of cell proliferation, differentiation and metabolism has been anticipated, but the evolution of our thinking and the analysis of such cross-talk have lagged behind the ever-expanding troupe of players and the recognition of multivalency as the rule, rather than the exception, in signalling biology. New insights have been provided by Kreuzer et al. in this issue of the Biochemical Journal, in which insulin is shown to provoke recruitment of Gαi-proteins to insulin-receptor-based complexes that can regulate the gain of insulin-receptor-catalysed autophosphorylation, a proximal point in the insulin-sensitive cascade of signalling. Understanding the convergence and cross-talk of signals from the receptor tyrosine kinases and G-protein-coupled receptor pathways in physical, spatial and temporal contexts will remain a major challenge of cell biology.
Glycogenesis and glucose oxidation during an intravenous glucose tolerance test in man
Authors: BLUCK Leslie J. C. , CLAPPERTON Allan T. , KIDNEY Cheryl V. , COWARD W. Andy
Published: June 2004

The quantity of deuterated glucose customarily given in labelled IVGTTs (intravenous glucose tolerance tests) changes the isotopic composition of the subject's body water enough to be detected by mass spectrometric techniques. Glucose undergoing direct glycogenesis does not contribute label to the body water pool, and isotope incorporated into it must have come from glucose that has either been oxidized or undergone indirect glycogenesis. By subtracting the amount of label found in body water from the total amount of glucose utilized, as calculated from the minimal model of glucose disappearance, it should be possible to study the partitioning of the dose given between direct glycogenesis in skeletal muscle and other metabolic pathways. To establish these principles, we used isotope ratio MS to determine body water composition in groups of healthy (n=7; mean weight, 76 kg; fasting plasma glucose and insulin, 5.1 mmol and 40 pmol respectively) and Type II diabetic (n=5; mean weight, 84 kg; fasting plasma glucose and insulin, 6.2 mmol and 75 pmol respectively) subjects undergoing IVGTTs. It was found that, for healthy subjects, 31% of the dose given was utilized in direct glycogenesis and this was decreased to 15% in diabetes. Defects in muscle glycogen synthesis in diabetes of the same order are well known from magnetic resonance studies. We conclude that measurement of label incorporation into body water is potentially useful for investigation of the metabolism of a glucose load in vivo during an IVGTT.
Acute Escherichia coli endotoxaemia decreases the plasma l-arginine/asymmetrical dimethylarginine ratio in humans
Authors: MITTERMAYER Friedrich , NAMIRANIAN Khodadad , PLEINER Johannes , SCHALLER Georg , WOLZT Michael
Published: June 2004

Acute inflammation impairs vascular function. Based on the association between endothelial dysfunction and plasma concentrations of L-arginine and the endogenous nitric oxide synthase inhibitor ADMA (asymmetrical dimethylarginine), we hypothesized that the ratio between L-arginine and ADMA could be affected by experimental inflammation. Plasma concentrations of L-arginine, ADMA and SDMA (symmetrical dimethylarginine) were studied at baseline and 3.5 h after intravenous administration of Escherichia coli endotoxin [LPS (lipopolysaccharide), 20 units/kg of body mass; n=8] or placebo (n=9) in healthy males. L-Arginine and dimethylarginines were quantified after solid-phase extraction by reversed-phase HPLC. Body temperature, heart rate and leucocyte count increased after LPS administration (P<0.01 for all). LPS administration decreased plasma concentrations of L-arginine from 66 µmol/l [95% CI (confidence interval): 56, 88] at baseline to 48 µmol/l (CI: 40, 60) after 3.5 h (P<0.02), but did not affect ADMA and SDMA concentrations. Consequently, the L-arginine/ADMA ratio declined significantly from a median of 159 (CI: 137, 193) to 135 (CI: 103, 146); a decrease of 25 (CI: -68, -13; P<0.02). L-Arginine, ADMA, SDMA and the L-arginine/ADMA ratio remained constant over time in controls. Acute inflammation reduces the L-arginine/ADMA ratio which could contribute to impaired vascular function.
Increased arterial stiffening and thickening in the paretic lower limb in patients with hemiparesis
Authors: OKABE Reiko , INABA Masaaki , SAKAI Shinrei , ISHIMURA Eiji , MORIGUCHI Atsushi , SHOJI Tetsuo , NISHIZAWA Yoshiki
Published: June 2004

Atherosclerosis has two key components, thickening and stiffening of arterial wall. These parameters are quantified ultrasonographically by IMT (intima-media thickness) and PWV (pulse wave velocity). In the present study, we determined the FA IMT (IMT of the bilateral femoral artery) and PWV of femoral–ankle (PWV fa) and brachial–ankle (PWV ba) segments in order to examine whether the degree of atherosclerosis is different between paretic and non-paretic lower limbs in 24 patients with hemiparesis. The values of PWV fa, PWV ba and FA IMT were all significantly greater on the paretic than the non-paretic side. Furthermore, significant decreases in masses of muscle, bone and fat, determined by dual-energy X-ray absorptiometry, were observed in paretic lower limbs compared with the non-paretic side. PWV fa correlated significantly and negatively with muscle mass (r=-0.488, P=0.0004) and tended to correlate negatively with BMC (bone mineral content; r=-0.264, P=0.069) when statistical analyses were performed with the paretic and non-paretic sides together. Multiple regression analysis elucidated that the muscle mass was associated significantly with PWV fa and PWV ba, independent of age, duration after cerebrovascular accident, gender, bone and fat mass and FA IMT. The muscle mass was still associated with increased PWV fa and PWV ba when multivariate analysis was conducted independently in the paretic and non-paretic sides. In summary, our results indicated that arterial thickening and stiffening were greater on the paretic than the non-paretic side and suggested that a decrease of muscle mass might be associated with increased arterial stiffening in the paretic lower limb.
Inspiratory flow limitation in obstructive sleep apnoea patients
Authors: FARRÉ Ramon
Published: June 2004

Patients suffering from the obstructive sleep apnoea syndrome (OSAS) experience nocturnal episodes of upper airway obstruction resulting in recurrent oxygen desaturations and arousals. Methods to quantify the nocturnal obstructive events are of interest for characterizing this prevalent sleep disorder. In a study published in this issue of Clinical Science, Bloch and co-workers propose the computation of a new index for objectively quantifying the degree of flow limitation in patients with OSAS. The results obtained in a bench test and in a pilot study in patients suggest that the flow limitation index proposed may help to better characterize the disturbed breathing events undergone by patients with OSAS.
Sympathetic overactivity in ischaemic heart disease
Authors: MALLIANI Alberto , MONTANO Nicola
Published: June 2004

The efficacy of pharmacological β-blockade in decreasing cardiac death in patients after myocardial infarction suggests the existence of sympathetic overactivity and indicates the importance of assessing its magnitude. The paper by Graham and co-workers in this issue of Clinical Science has attempted to address this issue by measuring muscle sympathetic nerve activity (MSNA) in various groups of patients and control subjects. It was found that, after myocardial infarction, there was sympathetic overactivity, which was more marked and more long-lasting than after unstable angina, whereas, in the presence of simple coronary artery disease, sympathetic activity did not differ from that in control subjects. Clear signs of sympathetic overactivity lasting for months after an acute myocardial infarction have already been reported using quite different methodology, i.e. spectral analysis of heart period and systolic arterial pressure variability. The soundest hypothesis to explain such a sympathetic overactivity appears to be based on the well-demonstrated finding that the ischaemic heart is a powerful site of origin of both excitatory and/or inhibitory reflexes, which may be of paramount clinical importance.
The strange eigenmode in Lagrangian coordinates
Authors: Thiffeault Jean-Luc
Published: June 2004

For a distribution advected by a simple chaotic map with diffusion, the “strange eigenmode” is investigated from the Lagrangian (material) viewpoint and compared to its Eulerian (spatial) counterpart. The eigenmode embodies the balance between diffusion and exponential stretching by a chaotic flow. It is not strictly an eigenmode in Lagrangian coordinates, because its spectrum is rescaled exponentially rapidly.
Strategic Asset Management in the Social Rented Sector: Approaches of Dutch and English Housing Associations
Authors: Gruis Vincent , Nieboer Nico , Thomas Andrew
Published: June 2004

In England and the Netherlands, in response to changes in housing policy and the market, there is widespread enthusiasm for asset management in the social rented sector. A key issue in both countries is the development of a strategic approach towards the formulation of asset management plans. The paper examines this issue in relation to the current practice of housing associations in both countries. Drawing on case studies in England and the Netherlands, it is concluded that in both countries practice varies widely between housing associations. Some of the approaches adopted by front-runners can be seen as models for other associations yet to develop a strategic approach towards asset management. Lessons can also be learnt across national boundaries, though necessarily mediated by differences between the two housing systems.
From Land Use Right to Land Development Right: Institutional Change in China's Urban Development
Authors: Zhu Jieming
Published: June 2004

There has been a fundamental institutional change during the transformation from the centrally controlled system to a market-oriented economy in China since the early 1980s. The socialist land use right as an institution of the out-going central planning system became an obstacle to the emerging land market for urban redevelopment, although land leasing as a new institution had created a real estate market primarily for the development of greenfield sites. From a case study of a district in Shanghai, it is discovered that rapid urban land redevelopment since 1992 has been greatly facilitated by the informal institution of a land development right. The land development right unlocks the process of land redevelopment in the central city, while the existing land users' land use right is taken care of during the gradual transition. Land use efficiency is improved to a great extent through land redevelopment to uses of higher productivity. However, the informal and insecure institution of the land development right induces hasty land redevelopment. Assisted by a capricious land use planning regime, the land development right yields sub-optimal development. Clarification of property rights is the goal for further institutional change.
Progress and Problems in Reforming Urban Land Use and Development Regulation in the Russian Federation
Authors: Trutnev Eduard K. , Valletta William , Yakoubov Mikhail O.
Published: June 2004

As part of the post-Soviet transition of civil law and economic relations, cities of the Russian Federation have sought to reform the system of urban land use and development regulation. Based on changes in the laws on urban development and the Land Code, the reforms have followed two approaches. Some cities have adopted the system of 'zoning' to create pre-defined land uses and parameters of development which apply uniformly to all land parcels in each urban zone. Other cities have retained the system of 'site-specific' definition of uses and development parameters. It appears that the 'zoning' system is more responsive to the new conditions in which private individuals, enterprises and institutions initiate development projects and provide capital funding, in response to market conditions. This can be shown by looking at the process of project development in the city of Novgorod (the Great), where Novgorod's reforms appear to offer to developers/investors an enhanced legal position, a lower level of risk and simpler and more cost-effective methods of acquiring land and development permits. This favourable situation appears to have made the city one of the most attractive locations for private investment in the Russian Federation.
The Effect of Impact Fees on the Price of New Single-family Housing
Authors: Mathur Shishir , Waddell Paul , Blanco Hilda
Published: June 2004

This paper provides new evidence on the effects of impact fees on housing prices, using an inventory of single-family housing sale transactions in the 38 cities and towns within King County, Washington, for the period 1991-2000. Although the effect of impact fees on housing prices has been examined previously, earlier studies have been limited by methodological deficiencies. This paper examines the effect of impact fees on new housing and their differential effect on housing price based on the quality of housing, and finds that the effect of impact fees on the price of new housing is quite significant and elastic, raising new home prices by about 166 per cent of the amount of the fee. The increase is 358 per cent for the higher-quality homes and is statistically insignificant for the lower-quality homes.
Transit Mobility, Jobs Access and Low-income Labour Participation in US Metropolitan Areas
Authors: Sanchez Thomas W. , Shen Qing , Peng Zhong-Ren
Published: June 2004

While policy-makers assert that increased public transit mobility can positively affect employment status for low-income persons, there is little empirical evidence to support this theory. It is generally assumed that public transit can effectively link unemployed, car-less, persons with appropriate job locations—hence the call for more public transit services to assist moving welfare recipients to gainful employment. Thus far, the available evidence is anecdotal, while general patterns of transit access in relationship to labour participation remain relatively unexplored. This analysis examines whether increased transit access is associated with the case status (employment status) of Temporary Assistance for Needy Families (TANF) recipients in the Atlanta, Georgia; Baltimore, Maryland; Dallas, Texas; Denver, Colorado; Milwaukee, Wisconsin ; and Portland, Oregon metropolitan areas. Individual TANF recipient location data, transit route/stop data and employment location data were used in limited dependent variable regression analyses to predict the employment status of TANF recipients. The results of this analysis indicate that access to fixed-route transit and employment concentrations had virtually no association with the employment outcomes of TANF recipients in the six selected metropolitan areas.
Territorial Cohesion: Old (French) Wine in New Bottles?
Authors: Faludi Andreas
Published: June 2004

If finally accepted, the new concept of territorial cohesion could mean a formal planning role for the European Union. The paper traces the French roots of this concept. As other concepts in European integration, it is subject to multiple interpretations. The initial focus has been on regional economic development as such. At present, territorial cohesion is also held to mean (for example, in the White Paper on European Governance) the co-ordination of policies with an impact on one and the same territory. Originally adhering to a more interventionist approach to spatial planning, the French have learned to factor balanced and sustainable development, concerns of the so-called comprehensive integrated approach, into the equation. Germans, in turn, are seeing sense in the new French focus on 'services of general economic interest'. Experts from both countries agree on the need for a spatial framework for Community policies. Such a framework would look somewhat like the European Spatial Development Perspective, but as part of territorial cohesion policy as a shared responsibility of the Union and its member-states. This would vindicate ideas of the French pioneers of European spatial planning.
Equalising Votes but Enabling Bias: The Electoral Impact of the 1977 and 1999 Ward Boundary Reviews in London
Authors: Railings Colin , Johnston Ron , Thrasher Michael
Published: June 2004

The boundaries of electoral units in Britain are periodically reviewed in order to enact the democratic principle of 'one person, one vote, one value'. The Commission-led reviews of local ward boundaries in London in the late 1970s and late 1990s both successfully reduced the variance in elector-councillor ratios within individual London boroughs and thereby helped to remove any bias in election outcomes stemming from malapportionment. However, other factors such as the efficiency with which a party's vote was distributed, the impact of differing levels of turnout and the intervention and success of third parties all remained crucial in determining the precise relationship between votes cast and seats won. It is impossible to ensure that all votes are equal under the 'first-past-the-post' system because these other components of electoral bias are either not subject to review, or amenable to manipulation.
THE EFFECT OF ODOURS, IRRITANT VAPOURS, AND MENTAL WORK UPON THE BLOOD FLOW
Authors: Shields T. E.
Published: June 2004

The most important of this investigation has been the completion of various improvements in the construction and use of the plethysmograph, by means of which numerous errors attending the use of the instrument have been eliminated.

The results of the work show that all olfactory sensations, so far as they produce any effect through the vasomotor system, tend to diminish the volume of the arm, and therefore presumably cause a congestion of the brain. Whenever the stimulation occassions an increase in the volume of the arm, as sometimes happens, it seems to be due to acceleration of the heart rate, which, of course, tends also to increase the supply of blood to the brain. The of odours varies in extent with different individuals, and with the same individual at different times. It was most marked in subjects sensitive to odours. Irritant vapours, such as formic acid, have a marked effect in the same direction—that is, they cause a strong diminution in the volume of the arm. The experiments give no support to the view that pleasant sensations are accompanied by a diminution of the

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blood supply to the brain and unpleasant sensations by the reverse efeect. In all my experiments mental work caused a marked and prolonged diminution in the volume of the arm. This vasomotor effect was sometimes preceded by a transitory increase in the volume of the arm caused by acceleration of heart rate.
A STUDY OF TYPES OF RESPIRATORY MOVEMENTS
Authors: Fitz G. W.
Published: June 2004

1. Children of the two sexes differ very little in the character of their respiratory movements.

2. Between girls and women and boys and men there is little or no difference in respiratory type.

3. Childbearing does not permanently affect respiration.

4. The natural type of respiration, for both sexes is one in which the movement is fairly equally balanced between chest and abdomen, the abdominal being somewhat in excess.

5. In typical unconstricted individuals the chest contributes about the same bulk of air as does the abdomen.

6. Constricting dress causes preponderance of thoracic movement in ratio to its restriction of abdominal movement and to the sensitiveness of the nervous co-ordination.
ON THE INHIBITORY INFLUENCE OF EOSIN UPON SPORULATION
Authors: Noguchi Hideyo
Published: June 2004

Sporulation of B. anthracis, B. subtilis, B. cereus, B. ruminatus, B. mesentericus, B. anthracoides and B. megatherium does not take place in an agar medium containing eosin "Gelb" in a concentration exceeding 0.5 per cent. In a concentration of 0.1 per cent. most of these bacteria fail to produce spores. The greatest sensitiveness is shown by B. cereus and B. mesentericus. In a bouillon medium sporulation is likewise inhibited by eosin, but after a longer time—seven weeks or more—sporulation still occurs where the concentration of the dye equals one-tenth per cent.

Sporulation of B. tetani, B. anthracis symptomaticus, B. botulismus, B. œdema maligni, B. enteritidis sporogenes, and B. putrificus does not take place in a medium containing eosin "Gelb" in concentrations exceeding 0.03 per cent. With these organisms, no difference was noted in the final effect, depending on the medium employed. No permanent loss of power to produce spores ensues with the bacteria tested even after long sojourn in the eosinized media.

It may be stated that on the whole the inhibitory action of eosin was more pronounced upon the anaerobic than upon the aerobic species of bacteria employed in these experiments.
TRANSPLANTATION IN MASS OF THE KIDNEYS
Authors: Carrel Alexis
Published: June 2004

Among so many etiological factors, it is impossible to discriminate which are responsible for the complications which took place in our experiments. An attempt to explain the occurrence of nephritis, cedema or calcification of the arterial system, for instance, will not be made, but the technique of the operations will be modified in order to suppress as much as possible the causes which may originate these secondary changes. The purpose of this article was not to analyze minutely the physiological or pathological character of the functions of transplanted kidneys, but merely to ascertain whether these functions are efficiently reëstablished.

It is to be concluded that an animal which has undergone a double nephrectomy and the grafting of both kidneys from another animal can secrete almost normal urine with his new organs, and live in good health at least for a few weeks. This demonstrates that it is possible to reëstablish efficiently the functions of transplanted kidneys.