bluuebunny/crossref_metadata_2025 huggingface dataset:
Tasmania and the Secret Ballot
Authors: Newman Terry
Published: December 2004

Encyclopaedias and handbooks, etc, all present Tasmania's starting date for implementing the secret ballot as 1858. But this research note argues the correct date was two years earlier in 1856. Moreover, before this, Tasmania's early 1830s participation in the Australia–wide proballot campaign, and its Constitutional inclinations towards the ballot have also been overlooked. To correct this omission relevant background is presented which highlights the riotous behaviour of pre–ballot elections. This is followed by a summary concerning the ballot's starting dates for Victoria and South Australia. Having confirmed these basic facts, further evidence is presented regarding Tasmania's true application of the secret ballot, including citations from a rare copy of the state's 1856 Electoral Act. In short, this “rediscovery” means that Tasmania definitely deserves a new place in the history of Australia's famous democratic innovation.
Accountability for Performance in Local Government
Authors: Kluvers Ron
Published: December 2004

The nature of accountability for performance in the public sector is explored in this paper by attempting to answer three questions: (1) How important is accountability for performance? (2) Which of the stakeholders consider themselves accountable for organisational performance, including contracts? (3) Does information asymmetry between stakeholders in regard to performance information affect accountability? This paper reports on research undertaken in Victorian local government based on the responses to a questionnaire by councillors and tier one and two managers. The results of the survey indicate that there is an understanding of accountability for performance, that both managers and councillors are considered to be accountable for performance but that information asymmetry and accountability relationships are problematic.
Chronic but not resistant mania: a case report
Authors: Mendhekar D. N. , Srivastav P. K. , Jiloha R. C. , Awana S.
Published: December 2004

Objective:  There are many nuances of elevated mood states. At times, it becomes very difficult to diagnose a case of mania because of personality style of the patient superimposed on a cyclothymic or hyperthymic temperament. Such cases may become chronic if timely intervention is not received. We describe a case of chronic mania of 48 years of duration.

Method:  A case report.

Results:  A 65‐year‐old male was admitted with history of mania for around five decades. The illness commenced at prepubertal age and was continuous without any residual symptoms of note. During the course of illness, he got involved in many devastating socio‐politico‐legal complications. He showed good response to treatment and is currently maintained well without medication. In this case we also described some psychosocial features that may have been responsible for the chronicity of the illness.

Conclusion:  We highlighted the need for correctly identifying chronic mania, early intervention and increasing treatment awareness. This case was not treatment resistant, suggesting that chronicity per se should not denote poor outcome.
Dopamine transporter binding in Gilles de la Tourette syndrome: a [123I]FP‐CIT/SPECT study
Authors: Serra‐Mestres J. , Ring H. A. , Costa D. C. , Gacinovic S. , Walker Z. , Lees A. J. , Robertson M. M. , Trimble M. R.
Published: December 2004

Objective:  To investigate dopamine transporter binding in Gilles de la Tourette syndrome (GTS) with SPECT and [123I]FP‐CIT.

Method:  Ten neuroleptic naïve/free patients with GTS, and 10 age‐ and gender‐matched normal volunteers were studied. Subjects were clinically evaluated. GTS severity and affective symptoms were measured and the presence of GTS‐related behaviours were recorded.

Results:  The GTS group showed significantly higher binding in both caudate and putamen nuclei than the controls. No associations were found between striatal binding ratios and measures of affect or GTS‐related behaviours.

Conclusion:  Patients with GTS show higher striatal binding of FP‐CIT to the striatum in comparison with age‐ and gender‐matched control subjects, indicating that dopamine transporter abnormalities are involved in the pathophysiology of GTS. These abnormalities appear to be distributed across both caudate and putamen.
The Sociocultural Context of Condom Use Within Marriage in Rural Lebanon
Authors: Kulczycki Andrzej
Published: December 2004

Prevalence rates for condom use are low across the Middle East despite limited alternate contraceptive options and growing awareness of sexually transmitted diseases (STDs) and HIV/AIDS. Virtually no research exists to explain this situation and to guide policy development or effective interventions. An analysis of 25 focus‐group discussions with married women and men, interviews with service providers, and a survey of 589 women of childbearing age in a rural district of southern Lebanon reveal that a broad and complex set of Sociocultural factors influence condom use. In the study area, 7 percent of married women currently use condoms, 24 percent reported ever use, and inconsistent use is common. Condoms are preferred primarily for their lack of physiological side effects, five factors were found to impede method adoption and sustained use. These include various encumbering beliefs, reduced sexual pleasure, adverse experiences, gender‐related fears and tensions, and a residual social stigma attached to condoms. Several strategies are suggested to increase and improve condom use. These issues will assume greater importance as fertility declines further in the region, demand for contraception continues to increase, and STDs and HIV/AIDS become more deeply rooted, as has occurred in other parts of the world.
How Family Planning Ideas Are Spread Within Social Groups in Rural Malawi
Authors: Paz Soldan Valerie A.
Published: December 2004

Using data from in‐depth interviews conducted between March and June 2002 in a rural district of Malawi, this study explores how family planning attitudes and practices spread among members of social groups. Gender differentials are found in how people determine other group members’ practices: Men “knew” about such practices from their observations of others’ family size and child spacing, whereas women's knowledge was based on their conversations with other women. The discussion topics relating to family planning also varied by sex: Men spoke about the pros and cons of limiting family size, whereas women spoke in detail about types of contraceptive methods, where to get them, their side effects, and covert contraceptive use. For men and women, the main trigger for family planning discussions was gossip. Whereas, generally, women first heard about family planning at the hospital, men stated that their first source of information was the radio or health‐drama group.
Emotional wellbeing and violence among social and solitary risky single occasion drinkers in adolescence
Authors: Kuntsche Emmanuel N. , Gmel Gerhard
Published: December 2004

Objectives  To classify adolescents according to risky single occasion drinking (RSOD) and their level of social integration, and to test whether these groups (social non‐RSODs, social RSODs, solitary non‐RSODs, solitary RSODs) differ in terms of emotional well‐being and violence‐related variables.

Method  K‐means cluster and multiple logistic regression analyses were performed based on a cross‐sectional national representative sample of 3861 8th and 9th graders in Switzerland (mean age 15.3; SD = 0.88).

Results  Although RSODs in general appear to be more violent, social RSODs tend to be more violent than solitary RSODs. Although RSODs reveal a lower life satisfaction generally and tend to have more depressive moods, solitary RSODs are even less satisfied and more depressive. In addition, the latter tend to have lower self‐esteem and are more often victims of bullying.

Conclusions  RSODs are not a homogeneous group of adolescents and preventive efforts, such as competence‐enhancing and social resistance programmes, should be applied in accordance with the constellation of associated problems: solitary RSODs appear to be socially inhibited, depressive and often victims of bullying, whereas social RSODs appear to be socially accepted but are prone to be violent offenders.
IL-4 and IL-10 modulate autoimmune haemolytic anaemia in NZB mice
Authors: Youssef A-R , Shen C-R , Lin C-L , Barker R N , Elson C J
Published: December 2004

New Zealand Black (NZB) mice spontaneously develop autoimmune haemolytic anaemia (AIHA). Here the effect of injecting NZB mice with plasmids encoding IL-4 (pIL-4) or IL-10 (pIL-10) on NZB disease was tested. Both constructs delayed the development of anaemia as judged by increased haematocrit values as compared with controls, but neither altered the IgG1 to IgG2 red blood cell (RBC) bound autoantibody levels. The increased haematocrit value was associated temporally with increased RBC bound IgG in NZB mice treated with pIL-10, but not pIL-4. By contrast, up-regulation of splenic macrophage FcγRIIb2 mRNA was associated temporally with increased haematocrit values in NZB mice given pIL-4. However, no such increase occurred in NZB mice that inhaled a peptide containing a dominant T-cell epitope, although this treatment is known to bias the autoimmune response towards Th2 and to reduce the severity of anaemia. It is considered that IL-4 treatment, in part, ameliorates NZB anaemia by increasing the expression of the inhibitory FcγRIIb2 and thereby reducing the capacity of splenic macrophages to phagocytose autoantibody coated RBC, but that this mechanism does not explain the beneficial effects of the inhaled peptide.
Intracellular T lymphocyte cytokine profiles in the aqueous humour of patients with uveitis and correlation with clinical phenotype
Authors: Hill T , Galatowicz G , Akerele T , Lau C H , Calder V , Lightman S
Published: December 2004

This study aimed to investigate whether T cells in aqueous humour are different in different types of uveitis and correlate with clinical phenotype. Patients with clinically different types of uveitis, but all displaying active anterior uveitis, were phenotyped and samples of aqueous humour (AH) and peripheral blood (PB) collected. Cells from AH and PB were separated by centrifugation and by density gradient centrifugation (to obtain mononuclear cells PBMC), respectively. Cells were activated with PMA and ionomycin in the presence of Brefeldin A, stained for surface markers and intracellular cytokines, and analysed by flow cytometry. The cytokine profile was correlated with the clinical phenotype. Increased percentages of interleukin (IL)-10+-, but not interferon (IFN)-γ+ T lymphocytes were found in AH compared with PB in patients with acute anterior uveitis (AAU), FHC or chronic panuveitis (PU). There was a trend towards elevated levels of IL-10+ T cells in AH from patients with FHC compared with AH from acute uveitis and panuveitis patients. Increased levels of IL-10+ T cells in AH compared with PB were also found in samples from patients with isolated uveitis, but not those with associated systemic disease. Levels of cytokine-positive T cells were not associated with the use of topical steroids or to the severity of the anterior uveitis. While type I cytokine-producing T lymphocytes are present in AH during AU, the presence of increased proportions of IL-10+ T lymphocytes in AH from patients with uveitis may be indicative of an anti-inflammatory mechanism that may influence the type and course of ocular inflammation in these patients.
Acute stress reduces intraparenchymal lung natural killer cells via beta-adrenergic stimulation
Authors: Kanemi O , Zhang X , Sakamoto Y , Ebina M , Nagatomi R
Published: December 2004

There are lines of evidence that natural killer (NK) cells are sensitive to physical and psychological stress. Alterations in the immune system including NK cells are known to differ among tissues and organs. The effect of stress on the lung immune system, however, has not been well documented in spite of the fact that the lungs always confront viral or bacterial attacks as well as tumour cell metastasis. In this study, we intended to investigate the effect of restraint stress on lung lymphocytes including NK cells. C57BL/6 mice were exposed to 2 h restraint stress. The concentration of plasma epinephrine significantly rose immediately after the release from restraint as compared to home-cage control mice. Flow cytometric analysis revealed that the numbers of most lymphocyte subsets including NK cells were decreased in the lungs and blood but not in the spleen, immediately after restraint stress. Immunohistochemical examination revealed that the number of NK cells was decreased in the intraparenchymal region of the lungs, while the number of alveolar macrophages did not change. The decrease in the number of NK cells in the lungs and blood was reversed by the administration of propranolol, a nonselective beta adrenergic antagonist. Taken together, our findings suggest that acute stress reduces the number of intraparenchymal lung NK cells via activation of beta adrenergic receptors.
Lower expression of Th1-related cytokines and inducible nitric oxide synthase in mice with streptozotocin-induced diabetes mellitus infected withMycobacterium tuberculosis
Authors: Yamashiro S , Kawakami K , Uezu K , Kinjo T , Miyagi K , Nakamura K , Saito A
Published: December 2004

Diabetes mellitus is an important predisposing factor for tuberculosis. The aim of this study was to investigate the mechanism underlying this association using a murine model. Mice with streptozotocin-induced diabetes mellitus were prone to Mycobacterium tuberculosis infection, as indicated by increased numbers of live bacteria in lung, liver and spleen. In diabetic mice, the levels of IL-12 and IFN-γ in the lung, liver and spleen were lower than those in control animals on day 14 postinfection, while the opposite was true for IL-4 levels in the lung and liver. The expression pattern of inducible nitric oxide synthase (iNOS), in the two mice types was as for IL-12 and IFN-γ. In addition, peritoneal exudate cells obtained from diabetic mice produced lower amounts of IL-12 and NO than those from control mice, when stimulated in vitro with M. bovis BCG. Spleen cells from diabetic mice infected with M. tuberculosis produced a significantly lower amount of IFN-γ upon restimulation with purified protein derivatives (PPD) than those from infected nondiabetic mice. Interestingly, addition of high glucose levels (33 mM) to the cultures of PPD-restimulated spleen cells reduced the synthesis of IFN-γ only in diabetic mice, and not in nondiabetic mice. Finally, control of blood glucose levels by insulin therapy resulted in improvement of the impaired host protection and Th1-related cytokine synthesis. Our results suggest that the reduced production of Th1-related cytokines and NO account for the hampered host defense against M. tuberculosis infection under diabetic conditions.
Acquired but reversible loss of erythrocyte complement receptor 1 (CR1, CD35) and its longitudinal alteration in patients with severe acute respiratory syndrome
Authors: Wang F S , Chu F L , Jin L , Li Y G , Zhang Z , Xu D , Shi M , Wu H , Moulds J-M
Published: December 2004

This longitudinal study investigates the change of erythrocyte complement receptor (E-CR1) expression in patients with severe acute respiratory syndrome (SARS). Circulating E-CR1 expression was semiquantified by flow cytometric analyses in 54 SARS patients and in 212 healthy individuals as a control. Since E-CR1 expression is influenced by the genetic polymorphisms in the CR1 gene, a major genetic polymorphism located within intron 27 of the CR1 gene was simultaneously analysed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). The results showed that the expression level of E-CR1 (referred to as net fluorescence intensity values, NFI) was statistically correlated with the relevant genetic genotypes among the Chinese population including the healthy individuals (NFI: 5·14 ± 0·82, 3·57 ± 0·66 and 2·67 ± 0·32 for HH, HL and LL genotypes, respectively) and SARS patients (NFI: 3·52 ± 0·91 and 2·63 ± 0·70 for HH and HL genotypes, respectively). Interestingly, the expression density of E-CR1 was found to fall significantly during the initiation and progressive phases (weeks 1 and 2 after the disease onset) and gradually returned close to normal through their whole convalescent phase (beginning from weeks 2 or 3 to weeks 7 or 8) in SARS patients irrespective CR1 genotype. In conclusion, our findings, at least, suggest that E-CR1 is likely involved in immune pathogenesis of SARS disease.
Combining nonpathogenic strains of Fusarium oxysporum with sodium chloride to suppress fusarium crown rot of asparagus in replanted fields
Authors: Elmer W. H.
Published: December 2004

Nonpathogenic strains of Fusarium oxysporum (NPFO) were tested alone, and in combination with rock salt (NaCl), for their ability to suppress fusarium crown and root rot of asparagus in replanted fields. Five NPFO strains were first screened with and without NaCl in the glasshouse using autoclaved asparagus soil mixed with potting mix. Asparagus roots were treated with NPFO suspensions or water, then inoculated 1 week later with pathogenic F. oxysporum; then after 1 week pots received 100 mL 1% NaCl or water. After 12 weeks the NPFO strain CS‐20 increased root weight when compared with controls. Rock salt did not affect root growth. Root lesions were reduced relative to the control by all NPFO strains and by NaCl applications, but there were no interactions between NPFO and NaCl. The NPFO strains CS‐20, CWB 314 and CWB 318 were compared with no treatment in two 4‐year replanted field studies in Hamden, and Windsor, CT, USA. Crowns were treated with NPFO at planting, and NaCl was broadcast at 280 kg ha−1, and at 560 kg ha−1 in later years. CWB 318 reduced disease at both sites and increased marketable yield in Hamden. Treatment with NaCl increased yield by 29% over controls in Windsor, but did not increase yield in Hamden. Recovered root isolates of F. oxysporum categorized by heterokaryon tests revealed that, in the glasshouse, CS‐20 colonized roots better than other strains, while CWB 318 was the only strain found on roots from field plots after 4 years. Applications of NaCl did not hinder the suppressive ability of the nonpathogenic strains. The perennial nature of asparagus may require that nonpathogenic strains possess superior survival traits along with biocontrol traits.
Kinetic analysis of zymogen autoactivation in the presence of a reversible inhibitor
Authors: Wang Wei‐Ning , Pan Xian‐Ming , Wang Zhi‐Xin
Published: December 2004

Limited proteolysis is a highly specific irreversible process, which can serve to initiate physiological function by converting a precursor protein into a biologically active form. When the activating enzyme and the activated enzyme coincide, the process is an autocatalytic zymogen activation (i.e. reactions in which the zymogens serves as a substrate for the corresponding active enzyme). The activity of proteases is frequently regulated by the binding of specific protease inhibitors. Thus, to understand the biological regulation of proteolysis, one must understand the role of protease inhibitors. In the present study, a detailed kinetic analysis of autocatalytic reaction modulated by a reversible inhibitor is represented. On the basis of the kinetic equation, a novel procedure is developed to evaluate the kinetic parameters of the reaction. As an example of the application of this method, effects of acetamidine,p‐amidinobenzamidine and benzamidine on the autoactivation of trypsinogen by trypsin were studied.
Nucleocytoplasmic shuttling of STAT transcription factors
Authors: Meyer Thomas , Vinkemeier Uwe
Published: December 2004

The signal transducer and activator of transcription (STAT) proteins have initially been described as cytoplasmic proteins that enter the nucleus only after cytokine treatment of cells. Contrary to this assumption, it was demonstrated that STATs are constantly shuttling between nucleus and cytoplasm irrespective of cytokine stimulation. This happens both via carrier‐dependent as well as carrier‐independent transportation. Moreover, it was also recognized that cytokine stimulation triggers nuclear retention of dimeric STATs, rather than affecting the rate of nuclear import. In summary, it is increasingly being appreciated that STAT nucleocytoplasmic cycling determines the quality of cytokine signaling and also constitutes an important area for microbial intervention.
Complex transcriptional and translational regulation of iPLA2γ resulting in multiple gene products containing dual competing sites for mitochondrial or peroxisomal localization
Authors: Mancuso David J. , Jenkins Christopher M. , Sims Harold F. , Cohen Joshua M. , Yang Jingyue , Gross Richard W.
Published: December 2004

Membrane‐associated calcium‐independent phospholipase A2γ (iPLA2γ) contains four potential in‐frame methionine start sites (Mancuso, D.J. Jenkins, C.M. & Gross, R.W. (2000) J. Biol. Chem.275, 9937–9945), but the mechanisms regulating the types, amount and subcellular localization of iPLA2γ in cells are incompletely understood. We now: (a) demonstrate the dramatic transcriptional repression of mRNA synthesis encoding iPLA2γ by a nucleotide sequence nested in the coding sequence itself; (b) localize the site of transcriptional repression to the most 5′ sequence encoding the iPLA2γ holoprotein; (c) identify the presence of nuclear protein constituents which bind to the repressor region by gel shift analysis; (d) demonstrate the translational regulation of distinct iPLA2γ isoforms; (e) identify multiple novel exons, promoters, and alternative splice variants of human iPLA2γ; (f) document the presence of dual‐competing subcellular localization signals in discrete isoforms of iPLA2γ; and (g) demonstrate the functional integrity of an N‐terminal mitochondrial localization signal by fluorescence imaging and the presence of iPLA2γ in the mitochondrial compartment of rat myocardium. The intricacy of the regulatory mechanisms of iPLA2γ biosynthesis in rat myocardium is underscored by the identification of seven distinct protein products that utilize multiple mechanisms (transcription, translation and proteolysis) to produce discrete iPLA2γ polypeptides containing either single or dual subcellular localization signals. This unanticipated complex interplay between peroxisomes and mitochondria mediated by competition for uptake of the nascent iPLA2γ polypeptide identifies a new level of phospholipase‐mediated metabolic regulation. Because uncoupling protein function is regulated by free fatty acids in mitochondria, these results suggest that iPLA2γ processing contributes to integrating respiration and thermogenesis in mitochondria.
Radical‐induced oxidation of metformin
Authors: Khouri H. , Collin F. , Bonnefont‐Rousselot D. , Legrand A. , Jore D. , Gardès‐Albert M.
Published: December 2004

Metformin (1,1‐dimethylbiguanide) is an antihyperglycaemic drug used to normalize glucose concentrations in type 2 diabetes. Furthermore, antioxidant benefits have been reported in diabetic patients treated with metformin. This work was aimed at studying the scavenging capacity of this drug against reactive oxygen species (ROS) like ·OH and ‐free radicals. ROS were produced by gamma radiolysis of water. The irradiated solutions of metformin were analyzed by UV/visible absorption spectrophotometry. It has been shown that hydroxyl free radicals react with metformin in a concentration‐dependent way. The maximum scavenging activity was obtained for concentrations of metformin ≥ 200 µmol·L−1, under our experimental conditions. An estimated value of 107 L·mol−1·s−1 has been determined for the second order rate constant k(·OH + metformin). Superoxide free radicals and hydrogen peroxide do not initiate any oxidation on metformin in our in vitro experiments.
Escherichia coli cyclopropane fatty acid synthase
Authors: Courtois Fabienne , Guérard Christine , Thomas Xavier , Ploux Olivier
Published: December 2004

Escherichia coli fatty acid cyclopropane synthase (CFAS) was overproduced and purified as a His6‐tagged protein. This recombinant enzyme is as active as the native enzyme with a Km of 90 µm for S‐AdoMet and a specific activity of 5 × 10−2 µmol·min−1·mg−1. The enzyme is devoid of organic or metal cofactors and is unable to catalyze the wash‐out of the methyl protons of S‐AdoMet to the solvent, data that do not support the ylide mechanism. Inactivation of the enzyme by 5,5′‐dithiobis‐(2‐nitrobenzoic acid) (DTNB), a pseudo first‐order process with a rate constant of 1.2 m−1·s−1, is not protected by substrates. Graphical analysis of the inactivation by DTNB revealed that only one cysteine is responsible for the inactivation of the enzyme. The three strictly conserved Cys residues among cyclopropane synthases, C139, C176 and C354 of the E. coli enzyme, were mutated to serine. The relative catalytic efficiency of the mutants were 16% for C139S, 150% for C176S and 63% for C354S. The three mutants were inactivated by DTNB at a rate comparable to the rate of inactivation of the His6‐tagged wild‐type enzyme, indicating that the Cys responsible for the loss of activity is not one of the conserved residues. Therefore, none of the conserved Cys residues is essential for catalysis and cannot be involved in covalent catalysis or general base catalysis. The inactivation is probably the result of steric hindrance, a phenomenon irrelevant to catalysis. It is very likely that E. coli CFAS operates via a carbocation mechanism, but the base and nucleophile remain to be identified.
Enhanced peptide secretion by gene disruption of CYM1, a novel protease in Saccharomyces cerevisiae
Authors: Jønson Lars , Rehfeld Jens F. , Johnsen Anders H.
Published: December 2004

Saccharomyces cerevisiae is a widely used host in the production of therapeutic peptides and proteins. Here we report the identification of a novel endoprotease in S. cerevisiae. It is encoded by the CYM1 gene and is specific for the C‐terminus of basic residues of heterologously expressed peptides. Gene disruption of CYM1 not only reduced the intracellular proteolysis, but also enhanced the secretion of heterologously expressed peptides such as growth hormone, pro‐B‐type natriuretic peptide and pro‐cholecystokinin. Cym1p resembles metalloendoproteases of the pitrilysin family with the HXXEH(X)E(71–77) catalytic domain as seen in insulysin, nardilysin and human metalloprotease 1. It is a nuclear encoded protease that localizes to mitochondria without a hydrophobic N‐terminal signal sequence or a C‐terminal tail‐anchor. The protease does not require post‐translational processing prior to activation and it contains cytosolic activity that processes peptides designated for the secretory pathway prior to translocation into the endoplasmic reticulum.
Molecular identification of monomeric aspartate racemase from Bifidobacterium bifidum
Authors: Yamashita Tatsuyuki , Ashiuchi Makoto , Ohnishi Kouhei , Kato Shin'ichiro , Nagata Shinji , Misono Haruo
Published: December 2004

Bifidobacterium bifidum is a useful probiotic agent exhibiting health‐promoting properties and contains d‐aspartate as an essential component of the cross‐linker moiety in the peptidoglycan. To help understand d‐aspartate biosynthesis in B. bifidum NBRC 14252, aspartate racemase, which catalyzes the racemization of d‐ and l‐aspartate, was purified to homogeneity and characterized. The enzyme was a monomer with a molecular mass of 27 kDa. This is the first report showing the presence of a monomeric aspartate racemase. Its enzymologic properties, such as its lack of cofactor requirement and susceptibility to thiol‐modifying reagents in catalysis, were similar to those of the dimeric aspartate racemase from Streptococcus thermophilus. The monomeric enzyme, however, showed a novel characteristic, namely, that its thermal stability significantly increased in the presence of aspartate, especially the d‐enantiomer. The gene encoding the monomeric aspartate racemase was cloned and overexpressed in Escherichia coli cells. The nucleotide sequence of the aspartate racemase gene encoded a peptide containing 241 amino acids with a calculated molecular mass of 26 784 Da. The recombinant enzyme was purified to homogeneity and its properties were almost the same as those of the B. bifidum enzyme.
Solution structure of the active‐centre mutant I14A of the histidine‐containing phosphocarrier protein from Staphylococcus carnosus
Authors: Möglich Andreas , Koch Brigitte , Gronwald Wolfram , Hengstenberg Wolfgang , Brunner Eike , Kalbitzer Hans Robert
Published: December 2004

High‐pressure NMR experiments performed on the histidine‐containing phosphocarrier protein (HPr) from Staphylococcus carnosus have shown that residue Ile14, which is located in the active‐centre loop, exhibits a peculiarly small pressure response. In contrast, the rest of the loop shows strong pressure effects as is expected for typical protein interaction sites. To elucidate the structural role of this residue, the mutant protein HPr(I14A), in which Ile14 is replaced by Ala, was produced and studied by solution NMR spectroscopy. On the basis of 1406 structural restraints including 20 directly detected hydrogen bonds, 49 1HN‐15N, and 25 1HN‐1Hα residual dipolar couplings, a well resolved three‐dimensional structure could be determined. The overall fold of the protein is not influenced by the mutation but characteristic conformational changes are introduced into the active‐centre loop. They lead to a displacement of the ring system of His15 and a distortion of the N‐terminus of the first helix, which supports the histidine ring. In addition, the C‐terminal helix is bent because the side chain of Leu86 located at the end of this helix partly fills the hydrophobic cavity created by the mutation. Xenon, which is known to occupy hydrophobic cavities, causes a partial reversal of the mutation‐induced structural effects. The observed structural changes explain the reduced phosphocarrier activity of the mutant and agree well with the earlier suggestion that Ile14 represents an anchoring point stabilizing the active‐centre loop in its correct conformation.
Trans‐splicing of a mutated glycosylasparaginase mRNA sequence by a group I ribozyme deficient in hydrolysis
Authors: Lundblad Eirik W. , Haugen Peik , Johansen Steinar D.
Published: December 2004

RNA reprogramming represents a new concept in correcting genetic defects at the RNA level. However, for the technique to be useful for therapy, the level of reprogramming must be appropriate. To improve the efficiency of group I ribozyme‐mediated RNA reprogramming, when using the Tetrahymena ribozyme, regions complementary to the target RNA have previously been extended in length and accessible sites in the target RNAs have been identified. As an alternative to the Tetrahymena model ribozyme, the DiGIR2 group I ribozyme, derived from a mobile group I intron in rDNA of the myxomycete Didymium iridis, represents a new and attractive tool in RNA reprogramming. We reported recently that the deletion of a structural element within the P9 domain of DiGIR2 turns off hydrolysis at the 3′ splice site (side reaction) without affecting self‐splicing [Haugen, P., Andreassen, M., Birgisdottir, Å.B. & Johansen, S.D. (2004) Eur. J. Biochem. 271, 1015–1024]. Here we analyze the potential of the modified ribozyme, deficient in hydrolysis at the 3′ splice site, for application in group I ribozyme‐mediated trans‐splicing of RNA. The improved ribozyme catalyses both cis‐splicing and trans‐splicing in vitro of a human glycosylasparaginase mRNA sequence with the same efficiency as the original DiGIR2 ribozyme, but without detectable levels of the unwanted hydrolysis.
Total chemical synthesis and NMR characterization of the glycopeptide tx5a, a heavily post‐translationally modified conotoxin, reveals that the glycan structure is α‐d‐Gal‐(1→3)‐α‐d‐GalNAc
Authors: Kang James , Low William , Norberg Thomas , Meisenhelder Jill , Hansson Karin , Stenflo Johan , Zhou Guo‐Ping , Imperial Julita , Olivera Baldomero M. , Rigby Alan C. , Craig A. Grey
Published: December 2004

The 13‐amino acid glycopeptide tx5a (Gla‐Cys‐Cys‐Gla‐Asp‐Gly‐Trp*‐Cys‐Cys‐Thr*‐Ala‐Ala‐Hyp‐OH, where Trp* = 6‐bromotryptophan and Thr* = Gal‐GalNAc‐threonine), isolated from Conus textile, causes hyperactivity and spasticity when injected intracerebral ventricularly into mice. It contains nine post‐translationally modified residues: four cysteine residues, two γ‐carboxyglutamic acid residues, and one residue each of 6‐bromotryptophan, 4‐trans‐hydroxyproline and glycosylated threonine. The chemical nature of each of these has been determined with the exception of the glycan linkage pattern on threonine and the stereochemistry of the 6‐bromotryptophan residue. Previous investigations have demonstrated that tx5a contains a disaccharide composed of N‐acetylgalactosamine (GalNAc) and galactose (Gal), but the interresidue linkage was not characterized. We hypothesized that tx5a contained the T‐antigen, β‐d‐Gal‐(1→3)‐α‐d‐GalNAc, one of the most common O‐linked glycan structures, identified previously in another Conus glycopeptide, contalukin‐G. We therefore utilized the peracetylated form of this glycan attached to Fmoc‐threonine in an attempted synthesis. While the result‐ing synthetic peptide (Gla‐Cys‐Cys‐Gla‐Asp‐Gly‐Trp*‐Cys‐Cys‐Thr*‐Ala‐Ala‐Hyp‐OH, where Trp* =6‐bromotryptophan and Thr* = β‐d‐Gal‐(1→3)‐α‐d‐GalNAc‐threonine) and the native peptide had almost identical mass spectra, a comparison of their RP‐HPLC chromatograms suggested that the two forms were not identical. Two‐dimensional 1H homonuclear and 13C‐1H heteronuclear NMR spectroscopy of native tx5a isolated from Conus textile was then used to determine that the glycan present on tx5a indeed is not the aforementioned T‐antigen, but rather α‐d‐Gal‐(1→3)‐α‐d‐GalNAc.
Solution structure of long neurotoxin NTX‐1 from the venom of Naja naja oxiana by 2D‐NMR spectroscopy
Authors: Talebzadeh‐Farooji Mehdi , Amininasab Mehriar , Elmi Maryam M. , Naderi‐Manesh Hossein , Sarbolouki Mohammad N.
Published: December 2004

The NMR solution structures of NTX‐1 (PDB code 1W6B and BMRB 6288), a long neurotoxin isolated from the venom of Naja naja oxiana, and the molecular dynamics simulation of these structures are reported. Calculations are based on 1114 NOEs, 19 hydrogen bonds, 19 dihedral angle restraints and secondary chemical shifts derived from 1H to 13C HSQC spectrum. Similar to other long neurotoxins, the three‐finger like structure shows a double and a triple stranded β‐sheet as well as some flexible regions, particularly at the tip of loop II and the C‐terminal tail. The solution NMR and molecular dynamics simulated structures are in good agreement with root mean square deviation values of 0.23 and 1 Å for residues involved in β‐sheet regions, respectively. The overall fold in the NMR structure is similar to that of the X‐ray crystallography, although some differences exist in loop I and the tip of loop II. The most functionally important residues are located at the tip of loop II and it appears that the mobility and the local structure in this region modulate the binding of NTX‐1 and other long neurotoxins to the nicotinic acetylcholine receptor.
Assessment for the complicated occurrence of nuisance odours from phytoplankton and environmental factors in a eutrophic lake
Authors: Sugiura Norio , Utsumi Motoo , Wei Bin , Iwami Norio , Okano Kunihiro , Kawauchi Yukio , Maekawa Takaaki
Published: December 2004

Odours in waters used for various purposes, especially musty odours, is a worldwide problem. To establish an early warning system, the nuisance odour in an aquatic environment, the causative microorganisms and compounds, and the mechanism of odour in Lake Kasumigaura, the second biggest lake in Japan, were examined using in situ studies. In this study, the mechanism for the occurrence of musty odours was determined, and the level of musty odour was discriminated, based on 32 months of phytoplankton data from three sampling stations. The results indicate that the cell numbers of Phormidium tenue, Synedra and Ankistrodesmus were significantly related to the occurrence of 2‐methlisoborneol (MIB) and geosmin. Furthermore, the odour occurrence for both MIB and geosmin was effectively predicted and validated using environmental factors as explanatory variables, using multiple linear regression and artificial neural network technology.
The implications of introducing a large piscivore (Lates calcarifer) into a regulated northern Australian river (Lake Kununurra, Western Australia)
Authors: Morgan David L. , Rowland Andrew J. , Gill Howard S. , Doupé Rob G.
Published: December 2004

The potential impacts of introducing barramundi (Lates calcarifer) for the purpose of recreational fishing into Lake Kununurra, a tropical impoundment in the Kimberley region of Western Australia, are predicted by dietary comparisons with the resident fishes of the lake. Classification of the pooled dietary data identified five major feeding groups based on similarities in food items consumed. There was no significant dietary overlap between L. calcarifer and all species within the lake. The current study demonstrates that adult L. calcarifer fed primarily on teleosts and decopods, and are known to prey on the majority of the fish species found in Lake Kununurra. Although the introduction of L. calcarifer to Lake Kununurra has the potential to influence the resident fish community through competition (for food and habitat) and predation, it is likely that its effects will be minor. However, the lack of any data that would allow estimation of the likely survival of stocked L. calcarifer fry and fingerlings in the reservoir needs to be addressed. Such data are mandatory if a successful fishery is to be developed in the reservoir.
The management lessons learned from sediment remediation in the Detroit River – western Lake Erie watershed
Authors: Hartig John H. , Heidtke Thomas M. , Zarull Michael A. , Yu Bonnie
Published: December 2004

During the 1970s−1990s, considerable emphasis was placed on minimizing the inputs of polychlorinated biphenyls (PCBs) from active sources. In addition, between 1993 and 2001, ≈ $US130 × 106 was spent for sediment remediation within the western Lake Erie – Detroit River basin. In general, although PCB contamination of the Detroit River and Lake Erie declined significantly between the 1970s and mid‐1990s, it has remained fairly stable over the past 10 years. Control of PCBs and other contaminants at their source remains a primary imperative for action. Remediation of contaminated sediments is growing in importance, however, as greater levels of source control are achieved. From a sediment management perspective, it is estimated that between 1993 and 2001 a substantially higher mass of PCBs (over two orders of magnitude higher) was removed as a result of contaminated sediment remediation, as compared to navigational dredging of shipping channels. In addition, there is a strong and compelling rationale for moving expeditiously to remediate severely contaminated sediment while it is still relatively contained in a small geographical area. The cost of not acting in a timely manner might be to exacerbate environmental problems including increased deformities and reproductive problems in wildlife, delayed ecosystem recovery and increased costs, or even preclusion of future sediment remediation. Based on discussions at a United States of America–Canada workshop held in 2002, key management advice includes continued emphasis to be placed on remediating contaminated sediment hot spots (including evaluating the effectiveness of projects), integrated monitoring efforts to be focused on beneficial use restoration and a high priority to be placed on sustaining and building upon modelling efforts, in order to be able to accurately predict and evaluate ecosystem responses to remedial and preventive actions.
Recent randomised trials in colorectal disease
Authors: Lawes D. , Taylor I.
Published: December 2004

Randomised trials represent the ‘gold standard’ for surgical research and have an important impact on clinical management. We provide an overview of the randomised trials, specifically related to the practice of colorectal surgery that were published between January and December 2003.
Laparoscopic colorectal surgery in Great Britain and Ireland – Where are we now?
Authors: Harinath G. , Shah P. R. , Haray P. N. , Foster M. E.
Published: December 2004

Objective  We aimed to gather information from the members of the Association of Coloproctology of Great Britain and Ireland (ACPGBI) to assess trends in the current practice of laparoscopic colorectal surgery.

Methods  A postal questionnaire survey of the members of ACPGBI.

Results  The response rate was 37% (200/540). Only 45 surgeons currently perform laparoscopic colorectal work in Great Britain and Ireland mainly right hemicolectomy and laparoscopic stoma formation, of these about one third practiced laparoscopy for benign colorectal conditions only. The majority (68%) of surgeons had enough resources at their place of work, but further training seemed to be a major issue. Nearly 22% of surgeons had not had any formal training. Only 50% of surgeons trained their specialist registrars. The incidence of conversion rate was not different for benign or malignant conditions and also did not appear to be related to the duration of experience. Only four surgeons had noted port a site recurrence during the past 10 years. Seventy‐five percent (150/200) felt that laparoscopic colorectal work could be carried out safely in a District General Hospital.

Conclusion  Laparoscopic colorectal surgery was being performed by a small minority of members of the ACPGBI although more surgeons had started to work in this field in recent years. The main areas of concern appeared to be a wide variation in the range of experience as indicated by the number of operations performed and limited formal training for consultants.
The use of a balloon catheter to improve evaluation in anorectal manometry
Authors: Bright T. , Kapoor R. , Voyvodich F. , Schloithe A. , Wattchow D.
Published: December 2004

The squeeze pressure in the anal canal reflects the contribution of the external anal sphincter and is normally assessed manometrically by asking patients to contract their anal muscles. However, this is an artificial situation as normally the external sphincter contracts to retain rectal content. Some patients with normal anal sphincter anatomy and innervation record low squeeze pressures suggesting that the concept of voluntary squeeze is foreign. The aim of this study was to examine whether squeezing to retain a balloon mimics the physiology of defaecation more accurately.

Patients undergoing routine anorectal manometry testing had in addition the inflation of a balloon catheter to the volume of the first and sustained sensation to simulate a faecal bolus within the rectum. The patient was asked to retain it when the balloon was subjected to gentle traction, thus contracting their anal sphincter to prevent passage of the balloon. Squeeze pressure was measured in response to voluntary contraction, the pressure generated to retain the balloon, then voluntary contraction again.

Eighteen women and 2 men were tested. The median maximal squeeze pressures with the routine assessment was 131.0cmH2O. This increased to 210.0cmH2O when the patients attempted to retain the balloon and fell to 165.4cmH2O when patients were reassessed with voluntary squeeze postintervention. 15 of the patients improved their squeeze pressures with traction on the balloon.

External anal sphincter contraction is difficult for some patients to perform on request. With traction on a balloon catheter anal squeeze pressures improved in most patients. This indicates that many patients perform maximal anal squeeze pressures better once that muscle group has been tested in a more normal physiological function. This simple technique could improve the accuracy of anorectal manometry results and evaluation in a larger popluation of symptomatic patients is warranted.
Colorectal foreign bodies
Authors: Clarke D. L. , Buccimazza I. , Anderson F. A. , Thomson S. R.
Published: December 2004

Objective  A pictorial review of colorectal foreign bodies and their extraction.

Methods  A prospective data‐base and photographic record of patients who presented with retained colorectal foreign bodies at our institution has been maintained since 1995. Information regarding the foreign body, clinical presentation and extraction technique were documented.

Results  All 13 patients were male: age range 2–66 years. Seven were Caucasian, 4 African and 1 Asian. The foreign bodies included a penknife, an aerosol deodorant spray can, a blue plastic tumbler, a plastic bag containing two bank‐notes and some marijuana, a plastic packet containing fish hooks, a penlight torch, a broomstick, a battery powered vibrator, a primus stove, a cap of an aerosol can, a piece of wire, a piece of hosepipe wrapped with wire and an iron bar. They entered the alimentary tract for a variety of reasons; anal autoeroticism (3), concealment (2), attention seeking behaviour (3), accidental (1), assault (2) and to alleviate constipation (2). Plain radiographs accurately demonstrated the site of the foreign body in 8 patients. Extraction was at laparotomy in 2 patients with peritonitis and in 3 who required extraction by colotomy. In 7 patients who had transanal extraction, four required general anaesthesia to facilitate extraction and extraction was possible under conscious sedation in the emergency room in three. The remaining patient extracted the foreign body himself and presented to hospital with a rectal perforation.

Conclusion  The emergency room physician must confirm the presence of a rectal foreign body. Extraction in the emergency room is usually not possible and patient's with retained rectal foreign bodies should be referred to a colorectal surgeon.
The future of coloproctology in the UK: The trainees view
Authors: Goodfellow P. B. , Buchanan G. N. ,   the DUKES club
Published: December 2004

Introduction  Major changes are imminent in the mode of surgical training and the manner both ‘‘general’’ and ‘‘specialist’’ surgeons provide services. This is the first interactive survey of UK Coloproctology trainees.

Methods  At the 2004 DUKES Club (colorectal trainees) meeting an interactive digital‐media voting system enabled blinded assessment of members regarding training and organisational issues in Coloproctology.

Results  78% of trainees intended to be colorectal specialists. 92% thought a specialist qualification necessary, 90% believed it should be administered at the time of certificate of completion of training (CCT). Overall, 40%, 40% and 20% would pay £1000<, £1000–3000 and >£5000 respectively per year for adequate training. Over 80% thought low anterior resection, APER, major lower GI‐bleeding, pelvic floor, IBD and rectal cancer surgery should only be performed by specialists, and many thought colonic cancer surgery (46%), diverticular surgery (52%), and perianal sepsis (38%) management was a specialist necessity.

Conclusion  UK Colorectal trainees believe a specialist exam necessary, colorectal specialists should treat a defined group of conditions/cases and would pay for adequate training.