bluuebunny/crossref_metadata_2025 huggingface dataset:
The effect of support group on the mental health and quality of life for mothers with autistic children
Authors: Shu B.‐C. , Lung F.‐W.
Published: December 2004

Background  The purpose of this study was to explore the effect of support groups on the mental health and quality of life for mothers with autistic children.

Method  A quasi‐experimental pre‐post control group design was used in this study. The mothers had children without chronic diseases diagnosed as autistic based on Diagnostic and Statistical Manual of Mental Disorder‐Fourth Edition (DSM‐IV) and were their primary caregivers. The mothers were assigned to control or intervention groups based on their willingness. The 10 weeks of the support group programme served as an intervention. There were three waves of data collection: pre‐test, post‐test, and after 1 month follow‐up. A total of 27 mothers with autistic children were assigned into experimental and control groups. In total, there were eight mothers in the experimental group and 19 mothers served as the control group.

Results  The results of this study showed that mental health had not significantly improved in the intervention group compared to those in the control group at the end of the first month during follow‐up. However, the subjective well‐being and employment status had an effect on their mental health. Only the subjective well‐being had an effect on their quality of life.

Conclusion  According to the results of this study, recommendations included (1) regularly promulgating a caregiver support group, (2) the advancement of training skills within the group, and (3) upgrading and promoting nurses’ ability to lead support groups in the future.
Associations of social support and hardiness with mental health among mothers of adult children with intellectual disability
Authors: Ben‐Zur H. , Duvdevany I. , Lury L.
Published: December 2004

Background The study was conducted with mothers of adult children with developmental disabilities and had two aims: (1) to examine the mental health, resources and stress among mothers who keep their adult child at home vs. those who choose placement in a community arrangement; and (2) to assess the associations of mothers’ resources and stress with mental health.

Method A sample of 100 mothers (mean age 60.67 years) of adult children with intellectual disability, 50% of whom had been placed in a community arrangement were asked to complete questionnaires measuring mental health, stress, hardiness and social support.

Results The comparisons between mothers who had placed their adult child in a community arrangement and those who had kept their child at home showed no significant differences for most indicators. Mental health, stress, hardiness and social support were highly intercorrelated, as expected, and hierarchical regression analyses indicated independent effects of stress, hardiness and out‐of‐home placement on mental health.

Conclusions The findings suggest that strengthening personal and social resources of mothers of adult children with developmental disabilities may be beneficial for their mental health.
Treatment and management of challenging behaviours in congregate and noncongregate community‐based supported accommodation
Authors: Robertson J. , Emerson E. , Pinkney L. , Caesar E. , Felce D. , Meek A. , Carr D. , Lowe K. , Knapp M. , Hallam A.
Published: December 2004

Objectives  To compare the nature and prevalence of use of procedures employed to treat and manage challenging behaviours across two approaches to providing community‐based supported accommodation for people with intellectual disabilities (ID) and severe challenging behaviour: noncongregate settings where the minority of residents have challenging behaviour, and congregate settings where the majority of residents have challenging behaviour.

Setting  Community‐based supported accommodation for people with ID and challenging behaviour.

Design  Longitudinal matched groups design.

Main outcome measures  The nature and prevalence of use of procedures employed to treat and manage challenging behaviours. Observed and reported severity of challenging behaviours.

Results  Both types of settings were associated with low prevalence of use of behavioural technologies for the reduction of challenging behaviour (less than 15% of participants). In contrast, high proportions of participant received antipsychotic medication in both noncongregate (56%) and congregate (80%) settings. Congregate settings were associated with the increased use of physical restraint as a reactive management strategy, with over half of participants being in receipt of physical restraint by two or more members of staff.

Discussion  Changes in reported and observed challenging behaviour over a 10‐month period were slight. The use of evidence‐based behavioural technologies for the reduction of challenging behaviour may have led to better outcomes.
Staff judgements of responsibility for the challenging behaviour of adults with intellectual disabilities
Authors: Dagnan D. , Cairns M.
Published: December 2004

Background  This study examines the importance of staff judgements of responsibility for challenging behaviour in predicting their emotional and intended helping responses.

Methods  Sixty‐two carers completed questionnaires rating attributions of internality, stability and controllability, emotions of sympathy and anger, judgements of responsibility for the development of challenging behaviour and for its resolution and intended effort in helping in response to a scenario describing an aggressive behaviour.

Results  Results showed significant correlations between judgements of responsibility and attributions, emotions and intended effort in helping. Regression analysis showed that the best predictor of intended helping is the emotion of sympathy and that sympathy is best predicted by the attribution of internality, the judgement that people are not responsible for the development of challenging behaviour and the judgement that they are responsible for the resolution of the behaviour.

Conclusions  Judgements of responsibility predict emotional and intended behavioural responses of carers of people with intellectual disabilities and challenging behaviour. The results are discussed in relation to previous work on carer attributions in response to challenging behaviour. Implications for clinical work with carers are considered.
Effect of light intensity on linear shrinkage of photo‐activated composite resins during setting
Authors: INOUE K. , HOWASHI G. , KANETOU T. , MASUMI S. , UENO O. , FUJII K.
Published: December 2004

summary  The purpose of this investigation was to examine the effects of light intensity on linear shrinkage of photo‐activated composite resins during setting. The materials used were four commercially available photo‐activated composite resins. Three light‐irradiation instruments were selected and prepared so as to obtain four light intensities (200, 480, 800 and 1600 mW cm−2). The linear shrinkage during setting was examined 10 min after light irradiation using a trial balance plastometer, and the specimen thickness was 2·0 mm for all materials. The depth of cure was examined according to the test method described in the International Organization for Standardization (ISO/FDIS 4049: 2000(E)). In measuring the linear shrinkage 60 s from the start of light irradiation for 10 s, there was a significant correlation (r = 0·89–0·94) between the amount of linear shrinkage and the light intensity: an increase in light intensity produced a greater linear shrinkage. Furthermore, there was a significant correlation (r = 0·92–1·0) between the linear shrinkage and the irradiation time: an increase in irradiation time resulted in a greater linear shrinkage. Values of the depth of cure ranged from 1·69 to 3·75 mm.
Some flexural properties of a nylon denture base polymer
Authors: YUNUS N. , RASHID A. A. , AZMI L. L. , ABU ‐ HASSAN M. I.
Published: December 2004

summary  Nylon denture base material could be a useful alternative to poly (methyl methacrylate) (PMMA) in special circumstances such as patient allergy to the monomer. The aim of this study was to evaluate the flexural properties of a nylon denture base material (Lucitone FRS), a conventional compression‐moulded heat‐polymerized (Meliodent), a compression‐moulded microwave‐polymerized (Acron MC) and an injection‐moulded microwave‐polymerized (Lucitone 199) PMMA polymers. The effect of aldehyde‐free, oxygen releasing disinfectant solution (Perform®) on these properties was also investigated. The flexural modulus and the flexural strength were assessed with a three‐point bending test. Specimens were stored in water at a temperature of 37 °C for 30 days. For each material, half of the prepared specimens were randomly selected and immersed in the disinfectant 24 h prior to testing. Results were compared statistically at a confidence level of 95%. The result showed that in both the control and disinfected groups, the flexural modulus of nylon was significantly lower than the three PMMA polymers. The flexural strength of nylon was significantly lower than those of Meliodent and Acron MC but was comparable with Lucitone 199. A 24‐h immersion in the disinfecting solution increased the rigidity of nylon denture base material.
Effect of preparation convergence on retention and seating discrepancy of complete veneer crowns
Authors: CHAN D. C. N. , WILSON A. H. , BARBE P. , CRONIN R. J. , CHUNG C. , CHUNG K.
Published: December 2004

summary  The purpose of this study was to evaluate the retention and marginal seating discrepancy of complete veneer crown cemented with zinc phosphate cement using various preparation convergence designs. The method employed was that of cementing cast metal crowns onto 46 metal dies fabricated as complete veneer crown preparations with various convergence angles ranging from 0° to 70° and a shoulder finish line. The marginal discrepancy was calculated by measuring the change in crown height before and after cementation using an optical measuring microscope. The force required to remove the crowns from the dies in a vertical direction was determined using an Instron universal testing machine and recorded as retention. The increase in preparation convergence exhibited a wide variation of seating discrepancy between specimens, ranging from 4·58 ± 1·13 to 73·13 ± 78·32 μm. Significant tilting of crown was observed as the convergence angle of preparation increased. The retention values ranged from 4·03 ± 0·61 to 12·12 ± 0·33 MPa. The Pearson analysis revealed statistically significant correlations between preparation convergence and marginal discrepancy (r = +0·62), and retention (r = −0·91). Crown retention and marginal discrepancy were influenced by the preparation convergence design.
Roles of the CBF2 and ZAT12 transcription factors in configuring the low temperature transcriptome of Arabidopsis
Authors: Vogel Jonathan T. , Zarka Daniel G. , Van Buskirk Heather A. , Fowler Sarah G. , Thomashow Michael F.
Published: December 2004

The CBF cold response pathway has a prominent role in cold acclimation. The pathway includes action of three transcription factors, CBF1, 2 and 3 (also known as DREB1b, c and a, respectively), that are rapidly induced in response to low temperature followed by expression of the CBF‐targeted genes (the CBF regulon) that act in concert to increase plant‐freezing tolerance. The results of transcriptome profiling and mutagenesis experiments, however, indicate that additional cold response pathways exist and may have important roles in life at low temperature. To further understand the roles that the CBF proteins play in configuring the low temperature transcriptome and to identify additional transcription factors with roles in cold acclimation, we used the Affymetrix GeneChip® containing probe sets for approximately 24 000 Arabidopsis genes to define a core set of cold‐responsive genes and to determine which genes were targets of CBF2 and 6 other transcription factors that appeared to be coordinately regulated with CBF2. A total of 514 genes were placed in the core set of cold‐responsive genes, 302 of which were upregulated and 212 downregulated. Hierarchical clustering and bioinformatic analysis indicated that the 514 cold‐responsive transcripts could be assigned to one of seven distinct expression classes and identified multiple potential novel cis‐acting cold‐regulatory elements. Eighty‐five cold‐induced genes and eight cold‐repressed genes were assigned to the CBF2 regulon. An additional nine cold‐induced genes and 15 cold‐repressed genes were assigned to a regulon controlled by ZAT12. Of the 25 core cold‐induced genes that were most highly upregulated (induced over 15‐fold), 19 genes (84%) were induced by CBF2 and another two genes (8%) were regulated by both CBF2 and ZAT12. Thus, the large majority (92%) of the most highly induced genes belong to the CBF and ZAT12 regulons. Constitutive expression of ZAT12 in Arabidopsis caused a small, but reproducible, increase in freezing tolerance, indicating a role for the ZAT12 regulon in cold acclimation. In addition, ZAT12 downregulated the expression of the CBF genes indicating a role for ZAT12 in a negative regulatory circuit that dampens expression of the CBF cold response pathway.
Expression of GFP‐fusions in Arabidopsis companion cells reveals non‐specific protein trafficking into sieve elements and identifies a novel post‐phloem domain in roots
Authors: Stadler Ruth , Wright Kathryn M. , Lauterbach Christian , Amon Gabi , Gahrtz Manfred , Feuerstein Andrea , Oparka Karl J. , Sauer Norbert
Published: December 2004

Transgenic Arabidopsis plants were constructed to express a range of GFP‐fusion proteins (36–67 kDa) under the companion cell (CC)‐specific AtSUC2 promoter. These plants were used to monitor the trafficking of these GFP‐fusion proteins from the CCs into the sieve elements (SEs) and their subsequent translocation within and out of the phloem. The results revealed a large size exclusion limit (SEL) (>67 kDa) for the plasmodesmata connecting SEs and CCs in the loading phloem. Membrane‐anchored GFP‐fusions and a GFP variant targeted to the endoplasmic reticulum (ER) remained inside the CCs and were used as ‘zero trafficking’ controls. In contrast, free GFP and all soluble GFP‐fusions, moved from the CCs into the SEs and were subsequently translocated through the phloem. Phloem unloading and post‐phloem transport of these mobile GFP‐fusions were studied in root tips, where post‐phloem transport occurred only for the free form of GFP. All of the other soluble GFP‐fusion variants were unloaded and restricted to a narrow zone of cells immediately adjacent to the mature protophloem. It appears that this domain of cells, which has a peripheral SEL of about 27–36 kDa, allows protein exchange between protophloem SEs and surrounding cells, but restricts general access of large proteins into the root tip. The presented data provide additional information on phloem development in Arabidopsis in relation to the formation of symplasmic domains.
The Mainz Severity Score Index: a new instrument for quantifying the Anderson–Fabry disease phenotype, and the response of patients to enzyme replacement therapy
Authors: Whybra C , Kampmann C , Krummenauer F , Ries M , Mengel E , Miebach E , Baehner F , Kim K , Bajbouj M , Schwarting A , Gal A , Beck M
Published: December 2004

Anderson–Fabry disease (AFD) is an X‐linked disorder caused by deficient activity of the lysosomal enzyme α‐galactosidase A. The availability of enzyme replacement therapy (ERT) for this debilitating condition has led to the need for a convenient and sensitive instrument to monitor clinical effects in an individual patient. This study aimed to develop a scoring system – the Mainz Severity Score Index (MSSI) – to measure the severity of AFD and to monitor the clinical course of the disease in response to ERT. Thirty‐nine patients (24 males and 15 females) with AFD were assessed using the MSSI immediately before and 1 year after commencing agalsidase alfa ERT. Control data were obtained from 23 patients in whom AFD was excluded. The MSSI of patients with AFD was significantly higher than that of patients with other severe debilitating diseases. The MSSI indicated that, although more men than women had symptoms classified as severe, overall, the median total severity scores were not significantly different between male and female patients. One year of ERT with agalsidase alfa led, in all patients, to a significant (p < 0.001) reduction in MSSI score (by a median of nine points). This study has shown that the MSSI score may be a useful, specific measure for objectively assessing the severity of AFD and for monitoring ERT‐related treatment effects.
Current clinical selection strategies for identification of hereditary non‐polyposis colorectal cancer families are inadequate: a meta‐analysis
Authors: Kievit W , De Bruin JHFM , Adang EMM , Ligtenberg MJL , Nagengast FM , Van Krieken JHJM , Hoogerbrugge N
Published: December 2004

Present guidelines to identify hereditary non‐polyposis colorectal cancer (HNPCC) families are criticized for limitations in accuracy. The Amsterdam criteria I and II (AC I and AC II) are used to predict a germline mutation in one of the mismatch repair genes. In families not fulfilling the AC I and AC II criteria, individual indications to test cancer specimens for microsatellite instability (MSI) are guided by the Bethesda Guidelines (BG). Germline mutation testing is then performed in patients who conform to the BG and show MSI. We investigated the sensitivity and specificity of AC I, AC II, and BG. A meta‐analysis of studies on the value of the AC I and AC II criteria for predicting germline mutation, as well as a meta‐analysis of BG for the detection of MSI was performed. For the AC I, sensitivity varied from 54 to 91% and specificity varied from 62 to 84%. For the AC II, the pooled sensitivity was 78% and specificity ranged between 46 and 68%. Post‐test probabilities of a positive test result were 0.61 and 0.46 for the AC I and AC II, respectively. Post‐test probabilities of a negative test result were 0.17 and 0.21 for the AC I and AC II, respectively. For the BG, the pooled sensitivity was 89% and pooled specificity was 53%. Post‐test probability of a positive test result was 41%, and post‐test probability of a negative test result was 9%. The sensitivity and specificity of the Amsterdam criteria for predicting a germline mutation that causes HNPCC is not sufficient. The BG are useful for the detection of MSI in a group of patients suspected of having familial colorectal cancer (CRC), but sensitivity is very low in the total group of newly diagnosed CRC patients. Therefore, a new strategy is needed for the identification of HNPCC.
Utilization of prenatal genetic testing by Israeli Moslem women: a national survey
Authors: Sher C , Romano‐Zelekha O , Green Ms , Shohat T
Published: December 2004

The number of prenatal genetic tests that are being offered to women is constantly increasing. However, there is little national data as to who is performing the tests and the reasons for doing or not doing so. The aim of the study was to evaluate the proportion of Israeli Moslem women who perform the various prenatal genetic tests and the factors affecting the performance of these tests. A 2‐day survey was conducted in all the maternity departments in Israel based on a structured interview. Of the 242 women interviewed, 74.2% underwent the triple test (TT), 15.8% of the women older than 35 years who were eligible to take the test free of charge underwent amniocentesis and only 1.5% underwent fragile‐X (FX) carrier testing which costs $100 privately. In the stepwise regression analysis, having fewer children and having had a higher education affected the performance of the TT. None of the sociodemographic factors were associated with the performance of amniocentesis or FX carrier testing, but the sample sizes were small. The main reason reported by the women for not performing the TT and FX carrier testing was not being referred for testing. The main reason for not performing amniocentesis was that they chose not to perform the test (the majority of these women were religious or ultrareligious). Consideration should be given to providing first‐trimester prenatal diagnosis when termination of pregnancy in the Moslem population is more acceptable. In addition, consideration should be given to including state‐subsidized FX testing as there is a low rate of FX testing partly due to the cost. Primary care givers should be educated about the importance of prenatal genetic testing.
Cascade genetic screening for familial hypercholesterolemia
Authors: Leren TP
Published: December 2004

Familial hypercholesterolemia (FH) is caused by a mutation in the low‐density lipoprotein receptor gene and is characterized by hypercholesterolemia, xanthomas, and premature coronary heart disease. Heterozygotes typically have values for total serum cholesterol in the range of 7–15 mmol/l and efficient lipid‐lowering drug therapy is available. However, only approximately 20% of patients are diagnosed and less than 10% are being adequately treated. The most cost‐effective strategy to diagnose patients with FH is to screen close relatives of patients already diagnosed with FH. This is referred to as cascade genetic screening. This review focuses on organization of a cascade genetic screening program for FH as well as cost‐efficiency assessments, health benefits, possible adverse effects, and the screening of children. The author concludes that cascade genetic screening for FH leads to health benefits and is cost‐effective without causing psychological or social damage. Accordingly, national cascade genetic screening programs for FH should be part of ordinary health care.
Low prevalence of Connexin 26 (GJB2) variants in Pakistani families with autosomal recessive non‐syndromic hearing impairment
Authors: Santos RLP , Wajid M , Pham TL , Hussan J , Ali G , Ahmad W , Leal SM
Published: December 2004

The Pakistani population has become an important resource for research on autosomal recessive non‐syndromic hearing impairment (ARNSHI) due to the availability of large extended and highly consanguineous pedigrees. Here is presented the first report on the prevalence of gap junction beta‐2 (GJB2) variants in Pakistan. One hundred and ninety‐six unrelated Pakistani families with ARNSHI were recruited for a study on the genetics of NSHI. DNA sequencing of the GJB2 coding region was done on two affected individuals per family. Evolutionary conservation and predicted effect on the protein product were studied in order to hypothesize whether or not a variant was potentially deleterious. Homozygous putatively functional GJB2 variants were identified in 6.1% of families. None of the putatively functional GJB2 variants were observed in the compound heterozygous state. The six putatively causative variants noted were 231G > A(W77X), 71G > A(W24X), 167delT, 95G > A(R32H), 358–360delGAG(delE120), and 269T > C(L90P), with 231G > A(W77X) and 71G > A(W24X) being the most common. In addition, five benign polymorphisms, 380G > A(R127H), 457G > A(V153I), 493C > T(R165W), 79G > A(V27I), and 341 A > G(E114G), were identified within this population. In a few individuals, benign polymorphisms were observed to occur on the same haplotype, namely [457G > A(V153I); 493C > T(R165W)] and [79G > A(V27I); 341 A > G(E114G)]. The spectrum of GJB2 sequence variants in Pakistan may reflect shared origins of hearing impairment alleles within the Indian subcontinent. The high degree of consanguinity within Pakistan may have maintained the GJB2 prevalence at a much lower rate than within India and other populations.
Massive gastrointestinal hemorrhage due to rupture of a donor pancreatic artery pseudoaneurysm in a pancreas transplant patient
Authors: Green Bryan T , Tuttle‐Newhall Janet , Suhocki Paul , Smith Stephen R , Barry O'Connor J
Published: December 2004

Abstract: Enteric drainage of secretions by anastomosing the donor duodenum to the recipient's small bowel has become common in pancreatic transplantation. While it eliminates many problems, endoscopic access to the transplanted duodenum and pancreas is made difficult. After a pancreas kidney transplant, the patient presented with massive hematochezia. Upper and lower endoscopy revealed large amounts of red blood in the colon but no specific bleeding site. Mesenteric angiography was normal but pelvic angiography showed rapid extravasation of contrast from a pseudoaneurysm of the pancreatic transplant artery. This was successfully embolized with coils. To the best of our knowledge, this is the first case of massive gastrointestinal hemorrhage because of rupture of a pseudoaneurysm of the donor pancreatic artery in a pancreas transplant patient. We report this case and review our institution's experience with all forms of gastrointestinal bleeding in pancreas transplant patients.
The impact of pancreas transplantation on patient employment opportunities
Authors: Knight Richard J , Daly Lisa
Published: December 2004

Abstract: Background: A questionnaire study was designed to determine whether the quality of life benefit provided by receiving a kidney–pancreas (KP) transplant increased the incentive to return to work, compared with patients receiving a kidney alone (KA) transplant.

Methods: A confidential questionnaire was sent to 58 patients with insulin‐dependent diabetes mellitus who had functioning grafts at least 6 months post‐transplant. Thirty‐eight patients had received a KP transplant and 20 patients, a KA transplant.

Results: Eighty‐three percent of patients returned the questionnaire (87% of KP and 75% of KA patients [p: non‐significant (ns)]. The pre‐transplant demographic variables of age, gender, educational status, and need for dialysis, were equivalent between the KP and KA groups, respectively (p: ns). At the time of transplantation 39% of KP patients and 33% of KA patients were employed at least 20 h/wk (p: ns). However, post‐transplant significantly more KP patients (73%) were working compared with KA patients (27%, p: 0.04). Additionally, pre‐transplant employment was independently associated with post‐transplant work status (p: 0.01).

Conclusion: These data suggest that the improvement in quality of life associated with pancreas transplantation provide an added incentive for diabetics with renal insufficiency to seek employment.
Long‐term outcome of liver transplantation for autoimmune hepatitis
Authors: Vogel Arndt , Heinrich Eyk , Bahr Matthias J , Rifai Kinan , Flemming Peer , Melter Michael , Klempnauer Juergen , Nashan Björn , Manns Michael P , Strassburg Christian P
Published: December 2004

Abstract:  Background:  Liver transplantation is the final therapeutic option for about 10% of patients with autoimmune hepatitis (AIH) who do not respond to medical therapy. The aim of this study was to evaluate the long‐term outcome in serologically defined subgroups of AIH after transplantation.

Methods:  Pre‐ and post‐transplantation data of 28 patients with AIH transplanted between 1987 and 1999 were retrospectively analyzed and compared with 24 patients, who underwent liver transplantation because of Wilson's disease and glycogen storage disease type 1.

Results:  Serological analyses identified patients with AIH type 1 (n = 13), type 2 (n = 5), and type 3 (n = 10). The 5‐yr patient survival rate after liver transplantation was 78.2%, which was not significantly different from the control group. Six AIH patients and four control patients required re‐transplantation because of initial non‐function, chronic rejection or AIH recurrence. Patients transplanted for AIH (88%) had more episodes of acute rejection when compared with patients transplanted for genetic liver diseases (50%). Clinical and histological features of chronic rejection were present in four patients, which did not differ significantly from the controls. Recurrence of AIH was diagnosed in nine patients (32%) based upon the presence of autoantibodies, increased γ‐globulins, steroid dependency, and histological evidence of chronic hepatitis. These combined features were not found in any of the controls.

Conclusions:  Our data do not suggest that AIH subtypes influence prognosis after liver transplantation. Despite a high frequency of acute cellular rejection episodes and disease recurrence, transplantation for AIH has a 5‐yr survival rate, which does not differ from that observed in patients transplanted for genetic liver diseases.
Solution Conformation of the Pseudomonas Syringae Pv. Syringae Phytotoxic Lipodepsipeptide Syringopeptin 25‐A
Authors: Ballio Alessandro , Bossa Francesco , Di Giorgio Domenico , Di Nola Alfredo , Manetti Cesare , Paci Maurizio , Scaloni Andrea , Segre Anna Laura
Published: December 2004

Syringopeptin 25‐A is a phytotoxic amphiphilic lipodepsipeptide containing 25 amino acid residues, produced by some isolates of the plant pathogenic bacterium Pseudomonas syringae pv. syringae. Previous papers have reported its covalent structure and some of its biological properties. Attention has now been directed to define its conformation in solution, a structural feature regarded as important for understanding its possible role in the bacterial colonization of host plants, and its toxic action on the plant cell.

Here we report the stereochemistry of its amino acid components, the complete interpretation of the two‐dimensional NMR spectra and NOE data, and finally the structure obtained by computer simulations applying distance geometry and molecular dynamics procedures.

The conformation of syringopeptin 25‐A in aqueous solution includes three different structured regions interrupted by rigid 2,3‐dehydro‐2‐aminobutyric acid residues: a loop from residue 2 to 6, a helicoidal zone from 8 to 15, and the lactone ring from 18 to 25. The three‐dimensional structure of the lactone moiety is very similar to that of two previously studied bioactive lipodepsinonapeptides. Preliminary circular dichroism evidence of conformational variations in solution of trifluoroethanol, which simulates a membrane‐like environment, are also reported.
Effect of Inhibitors on the Coordination Geometries of Cadmium at the Metal Sites in Angiotensin‐I‐Converting Enzyme
Authors: Carvalho Eugenia , Göthe Per Olof , Bauer Rogert , Danielsen Eva , Hemmingsen Lars
Published: December 2004

Perturbed angular correlation of γ‐rays (PAC) spectroscopy has been used to investigate the angiotensin‐I‐converting enzyme (ACE) of rabbit lung. By substituting the zinc ions in ACE with excited 111mCd2+ ions, analysis of PAC spectra gave directly the percentage of cadmium ions bound to ACE. The result of the analysis was a dissociation constant of about 1 μM for the cadmium‐ACE complex, and a stoichiometry of two moles cadmium/mole enzyme. Cadmium binding is thus about two orders of magnitude weaker than zinc binding to ACE but two orders of magnitude stronger than cobalt binding. PAC spectra monitor the nuclear quadrupole interaction (NQI) for 111mCd. The NQI for ACE exhibits very low frequencies in the PAC spectra with a rather large spectral broadening. In the presence of the inhibitor ramiprilat, the frequencies increase but the spectral broadening is about the same as for ACE without inhibitor. When the inhibitor captopril is added, very high frequencies are obtained consistent with sulfur binding, but now with a narrower distribution of NQI's. A simple molecular orbital analysis of the obtained NQI's has been performed, using a coordination sphere of two His, one Glu residue and a solvent ligand, equivalent to the zinc ligands in thermolysin and carboxypeptidase. The calculated spectral parameters could be modelled with the measured parameters if the solvent ligand is H2O in free ACE, carboxylate from ramiprilat in the ACE‐ramiprilat complex and a mercapto group in the ACE‐captopril complex. The coordination geometry for cadmium carboxypeptidase obtained by X‐ray diffraction gives a calculated set of NQI parameters consistent with the measured parameters for cadmium in the captopril‐ACE complex using a mercapto group as the solvent ligand. However, for ACE and its complex with ramiprilat, a significant distortion of the cadmium geometry for carboxypeptidase A had to be adopted in order to calculate NQI's close to the experimental values.
Isolation and Structural Characterization of N‐Acetyl‐ and N‐Glycolylneuraminic‐Acid‐Containing GalNAc‐GD1a Isomers, IV4GalNAcIV3Neu5AcII3Neu5GcGgOse4Cer and IV4GalNAcIV3Neu5AcII3Neu5AcGgOse4Cer, from Bovine Brain
Authors: Casellato Riccardo , Brocca Paola , Li Su‐Chen , Li Yu‐Teh , Sonnino Sandro
Published: December 2004

A ganglioside preparation containing two structurally related minor gangliosides (Gg 1+2) was isolated from bovine brain ganglioside mixture and characterized. Treatment of 50 g ganglioside mixture with Clostridium perfrigens sialidase, followed by chromatography on DEAE‐Sepharose and silica gel columns, yielded 20 mg Gg 1+2.

By chemical analyses, 1H‐ and 13C‐NMR spectroscopy, enzymic hydrolyses using human β‐hexosaminidase A and clostridial sialidase, and TLC overlay with the conjugated cholera toxin B subunit, the two novel gangliosides Gg 1 and Gg 2 were identified to be: Gg 1, GalNAc‐GD1a(Neu5Ac/Neu5Gc), β‐GalNAc‐(1–4)‐[α‐Neu5Ac‐(2–3)]‐β‐Gal‐(1–3)‐β‐GalNAc‐(1–4)‐[α‐Neu5Gc‐(2–3)]‐β‐Gal‐(1–4)‐β‐Glc‐(1–1)‐Cer: Gg 2, GalNAc‐GD1a(Neu5Gc/Neu5Ac), β‐GalNAc‐(1–4)‐[α‐Neu5Gc‐(2–3)]‐β‐Gal‐(1–3)‐β‐GalNAc‐(1–4)‐[α‐Neu5Ac‐(2–3) ‐β‐Gal‐(1–4)‐β‐Glc‐(1–1)‐Cer. These two gangliosides contain the identical pentasaccharide backbone except that the substitution of the two sialic acids, Neu5Ac and Neu5Gc, are in the reversed position of the external and the internal Gal residues. Our analyses showed that the content of Gg 1 and Gg 2 were approximately 0.12% and 0.08%1, respectively, of the total brain ganglioside mixture.
Modulation of the Platelet‐Derived‐Growth‐Factor‐Induced Calcium Signal by Extracellular/Intracellular pH in Syrian Hamster Embryo Cells
Authors: Isfort Robert J. , Stuard Sharon B. , Cody David B. , Ridder Gregg M. , Leboeuf Robert A.
Published: December 2004

Studies have been performed to understand the interactions and the role which intracellular calcium and intracellular pH have in mediating mitogen‐stimulated cellular proliferation. Stimulation of Syrian hamster embryo (SHE) cells with the mitogen platelet‐derived growth factor A/B (PDGF) results in intracellular acidification and capacitative calcium entry involving the intracellular release of calcium via the inositol trisphosphate γ receptor calcium channel, followed by an extracellular influx of calcium through a dihydropyridine‐sensitive plasma membrane calcium channel. Chronic extracellular/intracellular acidification results in the inactivation of both these calcium channels due to slowly reversible protein alterations. Paradoxically, transient intracellular acidification, like that following PDGF stimulation, could not stimulate the activation of either calcium channel. In addition, even though intracellular calcium fluxes by themselves could intiate intracellular acidification, loss of the PDGF‐induced calcium signal did not result in the loss of the PDGF‐induced transient intracellular acidification. Importantly with regard to the role intracellular calcium and pH have in mediating the mitogenic signal leading lo cellular proliferation, chronic extracellular/intracellular acidification, which leads to a complete loss of the PDGF‐induced calcium signal, did not result in the loss of PDGF‐induced mitogenesis. These results indicate that the PDGF‐induced calcium signal is not essential for PDGF‐stimulated mitogenesis in Syrian hamster embryo cells. In contrast, blocking the PDGF‐induced transient intracellular acidification completely blocks PDGF‐induced mitogenesis, indicating that the mitogen‐induced transient intracellular acidification, unlike the intracellular calcium ion signal, is indispensible for cellular proliferation in Syrian hamster embryo cells.
Isolation and Characterisation of a Recombinant, Precursor form of Lysosomal Acid α‐Glucosidase
Authors: Fuller Maria , Van Der Ploeg Ans , Reuser Arnold J. J. , Anson Donald S. , Hopwood John J.
Published: December 2004

Glycogenosis type II (GSD II, Pompe disease) is an autosomal recessive lysosomal storage disease that results from a deficiency of acid α‐glucosidase (GAA). Patients with this disorder are unable to break down lysosomal glycogen, which consequently accumulates in the lysosome, To evaluate enzyme replacement therapy for GSD II patients, we have expressed human GAA cDNA in Chinese hamster ovary‐K1 cells utilising a vector that places the cDNA under the transcriptional control of the human polypeptide chain elongation factor 1α gene promoter. A clonal cell line that secreted precursor recomibinant GAA at approximately 18 mg · 1−1 day−1 was identified. The precursor recombinant GAA was purified to homogeneity, had a molecular mass of 110 kDa as measured by SDS/PAGE, and was shown to have pH optima and kinetic parameters similar to those of GAA purified from human tissues. The partial N‐terminal amino acid sequence of recombinant GAA conformed to that derived from the nucleotide sequence of the cloned cDNA. The recombinant enzyme was taken up by cultured fibroblasts and skeletal muscle cells from GSD II patients, and was shown to correct the storage phenotype. Endocytosed GAA was localised to the lysosome and showed evidence of intracellular processing to a more mature form. Activity levels increased up to twice the normal value and uptake was prevented if cells were cultured in the presence of mannose 6‐phosphate.
Structure of Glycopeptides Isolated from Bovine Milk Component PP3
Authors: Girardet Jean‐Michel , Coddeville Bernadette , Plancke Yves , Strecker Gérard , Campagna Sylvie , Spik Geneviève , Linden Guy
Published: December 2004

The heat‐stable acid‐soluble phosphoglycoprotein component PP3 was isolated from the bovine milk proteose peptone fraction by concanavalin A affinity chromatography. Glycopeptides were released by pronase digestion of the milk component PP3 and were subsequently separated by high‐pH anion‐ex‐change chromatography on Carbopac PA‐1. The primary structures of the glycan and peptide moieties of eight N‐glycopeptides have been established by combining methylation analysis, mass spectrometry, 400‐MHz 1H‐NMR spectroscopy, and peptide sequence analysis. All the analyzed fractions contained biantennary N‐acetyllactosamine‐type carbohydrate chains, some of them with a GalNAc(β1–4)GlcNAc or a NeuAc(α2–6)GalNAc(β1–4)GlcNAc group. This particular sequence did or did not replace the Gal(β–4)GlcNAc group usually found in most N‐linked glycans. Moreover, the sialylated Gal and GalNAc residues were only found on the Man(α1–3) antenna.
Lipid Interaction of the 37‐kDa and 58‐kDa Fragments of the Helicobacter Pylori Cytotoxin
Authors: Moll Gert , Papini Emanuele , Colonna Raffaele , Burroni Daniela , Telford John , Rappuoli Rino , Montecucco Cesare
Published: December 2004

Helicobacter pylori cytotoxin vacA (95 kDa) causes a vacuolar degeneration of epithelial cells. There is evidence that this protein toxin acts inside cells, and hence has to cross a cell membrane. This cytotoxin is frequently obtained as two fragments of 58 kDa (p58) and 37 kDa (p37) and it is available only in minute amounts. Here, its membrane interaction was studied with the two fragments, produced in Escherichia coli. Light scattering and energy transfer experiments show that p37 and p38 cause aggregation and fusion of small unilamellar lipid vesicles: only a reversible aggregation is induced at neutral pH, whereas at acid pH fusion also takes place. p58, but not p37, causes potassium efflux from liposomes and this occurs only at acid pH. Hydrophobic photolabelling with photoactivatable phosphatidylcholines inserted into liposomes shows that both fragments are labelled at neutral pH. The amount of labelling of the two fragments is much higher at acid pH, consistent with a further penetration into the hydrophobic core of the lipid bilayer. Tryptophan fluorescence measurements indicate that the two fragments undergo a pH‐driven conformational change. These data are consistent with cytotoxin entry in the cell cytosol via an intracellular acidic compartment.
High level cell‐free expression and specific labeling of integral membrane proteins
Authors: Klammt Christian , Löhr Frank , Schäfer Birgit , Haase Winfried , Dötsch Volker , Rüterjans Heinz , Glaubitz Clemens , Bernhard Frank
Published: December 2004

We demonstrate the high level expression of integral membrane proteins (IMPs) in a cell‐free coupled transcription/translation system using a modified Escherichia coli S30 extract preparation and an optimized protocol. The expression of the E. coli small multidrug transporters EmrE and SugE containing four transmembrane segments (TMS), the multidrug transporter TehA with 10 putative TMS, and the cysteine transporter YfiK with six putative TMS, were analysed. All IMPs were produced at high levels yielding up to 2.7 mg of protein per mL of reaction volume. Whilst the vast majority of the synthesized IMPs were precipitated in the reaction mixture, the expression of a fluorescent EmrE‐sgGFP fusion construct showed evidence that a small part of the synthesized protein ‘remained soluble and this amount could be significantly increased by the addition of E. coli lipids into the cell‐free reaction. Alternatively, the majority of the precipitated IMPs could be solubilized in detergent micelles, and modifications to the solubilization procedures yielded proteins that were almost pure. The folding induced by formation of the proposed α‐helical secondary structures of the IMPs after solubilization in various micelles was monitored by CD spectroscopy. Furthermore, the reconstitution of EmrE, SugE and TehA into proteoliposomes was demonstrated by freeze‐fracture electron microscopy, and the function of EmrE was additionally analysed by the specific transport of ethidium. The cell‐free expression technique allowed efficient amino acid specific labeling of the IMPs with 15N isotopes, and the recording of solution NMR spectra of the solubilized EmrE, SugE and YfiK proteins further indicated a correctly folded conformation of the proteins.
Tungsten‐containing aldehyde oxidoreductase of Eubacterium acidaminophilum
Authors: Rauh David , Graentzdoerffer Andrea , Granderath Katrin , Andreesen Jan R. , Pich Andreas
Published: December 2004

Aldehyde oxidoreductase of Eubacterium acidaminophilum was purified to homogeneity under strict anaerobic conditions using a four‐step procedure. The purified enzyme was present as a monomer with an apparent molecular mass of 67 kDa and contained 6.0 ± 0.1 iron, 1.1 ± 0.2 tungsten, about 0.6 mol pterin cofactor and zinc, but no molybdenum. The enzyme activity was induced if a molar excess of electron donors, such as serine and/or formate, were supplied in the growth medium compared to readily available electron acceptors such as glycine betaine. Many aldehydes served as good substrates, thus enzyme activity obtained with acetaldehyde, propionaldehyde, butyraldehyde, isovaleraldehyde and benzaldehyde differed by a factor of less than two. Kinetic parameters were determined for all substrates tested. Oligonucleotides deduced from the N‐terminal amino acid sequence were used to isolate the encoding aorA gene and adjacent DNA regions. The deduced amino acid sequence of the aldehyde oxidoreductase exhibited high similarities to other tungsten‐containing aldehyde oxidoreductases from archaea. Transcription of the aorA gene was monocistronic and started from a σ54‐dependent promoter. Upstream of aorA, the gene aorR is localized whose product is similar to σ54‐dependent transcriptional activator proteins and, thus, AorR is probably involved in the regulation of aorA expression.
Anti‐oxidant status in relation to lipoproteins, leptin and pro‐inflammatory cytokines in children with steroid‐sensitive nephrotic syndrome
Authors: ECE AYDIN , ATAMER YILDIZ , GÜRKAN FUAT , BILICI MEKI , KOÇYIĞIT YÜKSEL
Published: December 2004

Background:  Reactive oxygen species and cytokines are reported to play a role in the proteinuria of nephrotic syndrome. The aim of this study was to investigate indirect evidence of oxidant activity together with leptin, lipoproteins and pro‐inflammatory cytokines in children with steroid‐sensitive nephrotic syndrome.

Methods:  A total of 40 children with steroid‐sensitive nephrotic syndrome (20 with newly onset or relapse comprised group I and 20 in remission while receiving steroids comprised group II) and 20 sex and age matched healthy control children were included. The following indirect parameters of oxidant activity were determined: serum malondialdehyde, erythrocyte superoxide dismutase, catalase and whole‐blood‐reduced glutathione. Serum leptin, lipids and lipoproteins were also determined.

Results:  Similar glutathione, increased malondialdehyde levels and decreased superoxide dismutase and catalase activity were observed in group I patients compared with controls. There was no significant difference in these variables between group I and group II (P > 0.05). Tumour necrosis factor‐alpha and interleukin‐6 concentrations were similar in patients and controls. Concentrations of interleukin‐1beta and interleukin‐8 were higher in the active phase of nephrotics compared with controls (P < 0.05). Significant positive correlations were found between malondialdehyde and interleukin‐1beta, interleukin‐6, leptin and lipoprotein (a) (P < 0.05). There were significant negative correlations between anti‐oxidants and leptin, lipoprotein (a) and several cytokines (P < 0.05).

Conclusions:  Changes in the concentrations of malondialdehyde, superoxide dismutase, catalase and glutathione are compatible with increased amounts of oxidation in steroid‐sensitive nephrotic syndrome. Leptin and pro‐inflammatory cytokines may be related to excessive protein permeability in nephrotic syndrome.
Caveolins in the repair phase of acute renal failure after oxidative stress
Authors: PERCY CHRISTINE , WATERS MICHAEL J , GOBÉ GLENDA
Published: December 2004

SUMMARY:  Ischaemia‐reperfusion and toxic injury are leading causes of acute renal failure (ARF). Both of these injury initiators use secondary mediators of damage in oxygen‐derived free radicals. Several recent publications about ischaemia‐reperfusion and toxin‐induced ARF have indicated that plasma membrane structures called caveolae, and their proteins, the caveolins, are potential participants in protecting or repairing renal tissues. Caveolae and caveolins have previously been ascribed many functions, a number of which may mediate cell death or survival of injured renal cells. This review proposes possible pathophysiological mechanisms by which altered caveolin‐1 expression and localization may affect renal cell survival following oxidative stress.
Process for improving the integration of care across the primary and acute care settings in rural South Australia: asthma as a case study
Authors: Laurence Caroline Olivia Mary , Beilby Justin , Campbell Sharon , Campbell John , Ponte Lorenzo , Woodward Glenda
Published: December 2004

Objective:  To develop a process for improving the integration of care across the rural acute and primary care settings using asthma as a case study.

Methods:  Development of the process on the analysis of case note audit, survey, interviews and a workshop.

Setting:  A rural region of South Australia.

Results:  A work plan for improving general practitioner (GP)–hospital integration was developed that resulted from analysis of the defined problem, GPs and stakeholder involvement, communication between all stakeholders, provision of an incentive to bring all the stakeholders together, and identification of evidence‐based solutions.

Conclusions:  Managing chronic disease in a community requires the integration of care across the primary and acute care setting. To be successful, GP–hospital integration initiatives require stakeholder involvement, locally developed solutions, engagement of GPs, communication and a well‐developed plan. This project provides a process for achieving this.

What this paper adds:  The present study outlines a process to improve the management of asthma for a specific community group in a rural region. It also provides a process to improve GP–hospital integration, which can be applied to a range of settings and health conditions to improve patient care.