bluuebunny/crossref_metadata_2025 huggingface dataset:
Children with Down Syndrome: oral development and morphology after use of palatal plates between 6 and 18 months of age
Authors: Bäckman B. , Grevér‐Sjölander A‐C. , Holm A‐K. , Johansson I.
Published: August 2003

Objectives. The aim of this study was to describe oral development and morphology in 18‐month‐old children with Down syndrome (DS) treated with palatal plates in combination with structured communication and speech training. The aim is further to describe the design of the palatal plates, compliance in their use and to give a brief report of their effect on oral motor function and speech.

Sample and methods. Forty‐two children with DS were followed from ≤ 6 months of age until 18 ± 3 months old. In addition to language intervention, and oral motor and sensory stimulation provided by speech therapists for all children with DS in Sweden, palatal plates provided by dentists are included in the training programme. In the evaluation, the children in the project were compared with two control groups of children matched for age; one group of children with DS who had not been treated with palatal plates, and one group of children with normal development.

Results. Compared to the children with normal development, both groups of children with DS had fewer teeth erupted and a lower prevalence of sucking habits. Deviant morphology of the tongue in the form of diastase, lingua plicata or a sulcus in the anterior third of the tongue was only seen in children with DS. All children with normal development had positive values for overjet compared to 53% of the children with DS. The palatal plates were used 2–3 times daily for a total mean time of 15 min. Compliance in use of the plates decreased with age, mainly due to eruption of teeth and subsequent loss of retention. Evaluation of oral motor function and speech show that the children with DS in the project had better motor prerequisites for articulation than the control children with DS.

Conclusion. Palatal plate therapy did not affect oral parameters, i.e., eruption of teeth, types and prevalence of sucking habits, tongue morphology and symptoms of hypotonia. In combination with oral motor and sensory stimulation, palatal plate therapy had a positive effect on oral motor performance and prerequisites for articulation.
Perceived information needs in respect of orthodontics amongst 11–12‐year‐old girls: a study through health visitor sessions in schools
Authors: Habibian M. , Gelbier S. , Munday B. A.
Published: August 2003

Aim. The aims of this study were: to explore: (i) the knowledge and views regarding orthodontics of a group of 11–12‐year‐old girls attending a school in South‐east London and (ii) the terms that they used to obtain the information.

Methods and subjects. The study used Dental Health Education sessions to investigate the aims of the study. Eight DHE sessions at a secondary school for girls were tape recorded. In order to raise the issue of orthodontics and trigger the formation of questions during health education session, a worksheet containing true/false questions, a crossword puzzle regarding orthodontics and some open ended questions was designed and sent to students. They were required to read and complete the worksheet before each session. They were not required to return the completed worksheets to the investigators but did return them to their teachers. The sessions were tape recorded and supplemented by notes taken at the sessions by the investigator. A total of eight DHE sessions, attended by 14 girls each, were tape‐recorded. Each tape recording was immediately transcribed verbatim. The next stage was to organize the data and to single out the orthodontic questions and discussions and categorize them.

Results. A total of 117 girls aged 11–12‐year‐old comprised the study group: 77% were white and 23% black children. After reading the transcripts several times, certain themes on orthodontics emerged. The results showed that children questioned different aspects of orthodontics. Nine themes emerged from their questions and discussions. They wanted to know why orthodontic treatment was carried out and when was the right time to start treatment. They were very keen to find out the differences between different orthodontic appliances. The psychosocial impacts of wearing an orthodontic appliance, i.e., experience of pain as well as the need for extraction of some permanent teeth as part of the treatment were of concern. They asked some questions on the need for repair, adjustment and taking care of appliances. The aetiology of malocclusion was another theme that emerged. The students tended to ask questions and describe problems in their own lay terms.

Conclusion. The methodology used in this study provided an opportunity to assess the information needs with regards to orthodontics of a group of children attending a school in Southwark, London, UK. It was successful in discovering the views and concerns, and to some extent, their knowledge regarding orthodontics and the terms pupils used in asking questions and making comments.
The progression of tooth erosion in a cohort of adolescents of mixed ethnicity
Authors: Dugmore C. R. , Rock W. P.
Published: August 2003

Objectives. To establish the prevalence of tooth erosion in a sample of 12‐year‐old children and to monitor changes over the subsequent 2 years.

Methods. A random sample of 1753 children aged 12 years was drawn from all 62 state maintained schools in Leicestershire. A total of 1308 were re‐examined 2 years later. Erosion was recorded on incisors and first molars using an erosion index based upon that from the Children's Dental Health in the United Kingdom 1993 survey. A score was also allocated to each subject according to the most advanced lesion in the mouth.

Results. Erosion was present in 56·3% of subjects at age 12 and 64·1% at age 14. Deep enamel or dentine was eroded in 4·9% and 13·1% of subjects, respectively, at the same ages. One hundred and sixty‐one (12·3%) children who were erosion‐free at 12 years of age developed erosion over the subsequent 2 years. Boys had more erosion than girls, as did white compared to Asian children. Associations were found between erosion experience and social deprivation.

Conclusion. New erosive lesions developed in 12·3% of the subjects between the ages of 12 and 14 years. New or more advanced lesions were seen in 27% of the children over the 2 years of the study. Males, white children and social deprivation were significantly associated with erosion experience.
Bone metabolism in ochronotic patients
Authors: Aliberti G. , Pulignano I. , Schiappoli A. , Minisola S. , Romagnoli E. , Proietta M.
Published: August 2003

Abstract. Aliberti G, Pulignano I, Schiappoli A, Minisola S, Romagnoli E, Proietta M (Università di Roma ‘La Sapienza’, Rome, Italy). Bone metabolism in ochronotic patients (Case report). J Intern Med 2003; 254: 296–300.

We investigated skeletal involvement in five male and two female patients with ochronosis, aged 26–82 years. The main parameters of mineral metabolism, together with biochemical markers of bone resorption (urinary N‐telopeptides of type I collagen) and formation (serum bone isoenzyme of alkaline phosphatase and serum osteocalcin) were evaluated. In the same subjects lumbar spine and femoral bone mineral density (BMD) were also measured by dual energy X‐ray absorptiometry. All patients but the younger 26‐year‐old patient had lower than normal bone mass at femoral neck and total hip, showing marked osteopenia in three cases and osteoporosis in the remaining three cases. However, at lumbar spine BMD measurement provided spuriously overestimated results, because of intervertebral disc calcification and osteophyte formation. As far as biochemical markers of bone turnover are concerned, the most relevant finding was the increased N‐telopeptides of type I collagen urinary excretion. Our results suggest that ochronosis may be associated with increased bone resorption rate leading to an accelerated bone loss. A role of the homogentisic acid polymer deposit in bone matrix and cells, possibly with osteocyte damage and interference in collagen metabolism, might be hypothesized.
Hereditary transthyretin amyloidosis from a Scandinavian perspective
Authors: Suhr O. B. , Svendsen I. Hastrup , Andersson R. , Danielsson Å. , Holmgren G. , Ranløv P. J.
Published: August 2003

Abstract. Suhr OB, Svendsen IH, Andersson R, Danielsson Å, Holmgren G, Ranløv PJ (Umeå University Hospital, Umeå, Sweden; Rigshospitalet, København Ø, Denmark; Örnsköldsviks Hospital, Örnsköldsvik, Sweden; and Hørsholm Hospital, Hørsholm, Denmark). Hereditary transthyretin amyloidosis from a Scandinavian perspective (Review). J Intern Med 2003; 254: 225–235.

Hereditary transthyretin (TTR) amyloidosis is a rare often fatal form of systemic amyloidosis, that until recently was considered intractable, with the patients dying from the disease 5–15 years after onset. The phenotype of the disease varies according to the type of mutation, but generally the heart and/or the nervous system is affected. Liver and in some cases heart transplantation has now been shown to stop the progress of the disease, but the outcome depends on the patients’ status at the time of operation, as no substantial improvement of the patients’ symptoms has been noted after the procedure. Thus an early diagnosis is of importance for the outcome.

In the following, we summarize our knowledge of the amyloidogenic TTR mutations found in the Scandinavian countries, their symptoms, how to settle the diagnosis and the outcome of transplantation. Besides, the problems arising from our capability to genetically test asymptomatic members of affected families for the trait will be discussed.
Morbidity in cardiovascular diseases in immigrants in Sweden
Authors: Gadd M. , Johansson S.‐E. , Sundquist J. , Wändell P.
Published: August 2003

Abstract. Gadd M, Johansson S‐E, Sundquist J, Wändell P (Family Medicine Stockholm, Karolinska Institutet, Huddinge, Sweden). Morbidity in cardiovascular diseases in immigrants in Sweden. J Intern Med 2003; 254: 236–243.

Introduction. Although immigration to Sweden has increased in the last few decades, the incidence rates of cardiovascular disease and coronary heart disease in immigrants are unknown. The aim of the present study is to estimate whether place of birth affects the incidence rates of cardiovascular disease and coronary heart disease.

Material and methods. The study was designed as a follow‐up study on morbidity in cardiovascular disease and coronary heart disease between 1 January 1997 and 31 December 1998, including three and a half million persons with age range 35–64 years, of whom 550 000 were born abroad, from the database MigMed consisting of the whole Swedish population. Incidence rates and relative risks were estimated by indirect standardization and a proportional hazard model.

Results. The age‐adjusted risk of coronary heart disease was higher in most foreign‐born groups than in Swedes. For example, in nine of 12 male groups, the relative risks varied between 1.1 and 2.2, and in seven of 12 female groups, the relative risks varied between 1.4 and 2.5. When also adjusting for level of education and employment status, the risks were still high, but on a lower level.

Conclusions.  Foreign‐born people possess an over‐risk of cardiovascular or coronary heart disease(CVD/CHD) compared with Swedish‐born persons, also when level of education and employment status are taken into account.
Prognostic value of plasma C‐reactive protein and fibrinogen determinations in patients with acute myocardial infarction treated with thrombolysis
Authors: Bennermo M. , Held C. , Hamsten A. , Strandberg L.‐E. , Ericsson C.‐G. , Hansson L.‐O. , Tornvall P.
Published: August 2003

Abstract. Bennermo M, Held C, Hamsten A, Strandberg L‐E, Ericsson C‐G, Hansson L‐O, Tornvall P (Danderyd Hospital; The Atherosclerosis Research Unit at King Gustaf V Research Institute, Karolinska Hospital; and Norrtälje Hospital, Karolinska Institute, Stockholm, Sweden). Prognostic value of plasma C‐reactive protein and fibrinogen determinations in patients with acute myocardial infarction treated with thrombolysis. J Intern Med 2003; 254: 244–250.

Objectives. To investigate the prognostic value of plasma C‐reactive protein (CRP) and fibrinogen determinations in patients with acute myocardial infarction treated with thrombolysis.

Design. Longitudinal study of morbidity and mortality.

Setting. Coronary care unit at Danderyd Hospital, Stockholm, Sweden.

Subjects. A total of 222 patients aged 75 years or below, treated with thrombolysis because of typical symptoms of myocardial infarction and electrocardiogram showing ST‐segment elevation or bundle branch block were included in the study. The patients were followed for 24–60 months (mean 40 ± 16 months).

Main outcome measures. Cardiovascular death or new myocardial infarction.

Results. Concentrations of CRP were significantly higher at 48 h than at 3 months, whilst the levels of fibrinogen were similar. CRP and fibrinogen concentrations measured during the acute phase of myocardial infarction were associated with cardiovascular death or a new myocardial infarction during follow‐up in univariate analysis. CRP levels measured 3 months after the acute event were not associated with subsequent events whereas fibrinogen concentrations showed a borderline prognostic significance (P = 0.05). When CRP and fibrinogen were entered into multivariate analysis together with the previously established prognostic factors in the patient group (age, diabetes mellitus and left ventricular function), these markers of inflammation did not add further prognostic information.

Conclusion. C‐reactive protein and fibrinogen do not carry the same independent prognostic information after acute myocardial infarction treated with thrombolysis as in studies previously reported for patients with unstable angina or non‐Q‐wave myocardial infarction.
Increasing body mass index at diagnosis of diabetes in young adult people during 1983–1999 in the Diabetes Incidence Study in Sweden (DISS)
Authors: Littorin B. , Nyström L. , Gullberg B. , Råstam L. , Östman J. , Arnqvist H. J. , Björk E. , Blohmé G. , Bolinder J. , Eriksson J. W. , Scherstén B. , Sundkvist G.
Published: August 2003

Abstract.  Littorin B, Nyström L, Gullberg B, Råstam L, Östman J, Arnqvist HJ, Björk E, Blohmé G, Bolinder J, Eriksson JW, Scherstén B, Sundkvist G (University of Malmö/Lund, Malmö/Lund; Umeå University, Umeå; University Hospital, Uppsala; Söder Hospital, Stockholm; Huddinge Hospital, Huddinge; Faculty of Health Sciences, University of Linköping, Linköping and University Hospital, Umeå; Malmö University Hospital, Malmö, Sweden). Increasing body mass index at diagnosis of diabetes in young adult people during 1983–1999 in the Diabetes Incidence Study in Sweden (DISS). J Intern Med 2003; 254: 251–256.

Objective.  To study trends in body mass index (BMI) at diagnosis of diabetes in all young Swedish adults in the age range of 15–34 years registered in a nation‐based registry.

Design.  The BMI was assessed at diagnosis in diabetic patients 15–34 years of age at diagnosis, for a period of 17 years (1983–1999). Islet cell antibodies (ICA) were measured during three periods (1987–1988, 1992–1993 and 1998–1999).

Setting.  A nationwide study (Diabetes Incidence Study in Sweden).

Subjects.  A total of 4727 type 1 and 1083 type 2 diabetic patients.

Main outcome measures.  Incidence‐year specific BMI adjusted for age, gender and time of diagnosis (month).

Results.  Body mass index at diagnosis increased significantly both in type 1 (21.4 ± 3.6 to 22.5 ± 4.0; P < 0.0001) and in type 2 (27.4 ± 6.8 to 32.0 ± 6.0; P < 0.0001) diabetic patients, also when adjusted for age, gender and month of diagnosis. A similar significant increase in BMI was found in type 1 diabetic patients and in type 2 diabetic patients in the periods 1987–1988, 1992–1993 and 1998–1999; years when ICA were assessed and considered in the classification of diabetes. Despite this increase in BMI, there was no increase in the incidence of diabetes in young‐adult people in Sweden.

Conclusion.  Body mass index at diagnosis of diabetes in subjects 15–34 years of age has substantially increased during 1983–1999 in Sweden when adjusted for age, gender and month of diagnosis.
Impact of clinical experience on quantification of clinical signs at physical examination
Authors: Drager L. F. , Abe J. M. , Martins M. A. , Lotufo P. A. , Benseñor I. J. M.
Published: August 2003

Abstract.  Drager LF, Abe JM, Martins MA, Lotufo PA, Benseñor IJM (School of Medicine, University of São Paulo, São Paulo, Brazil). Impact of clinical experience on quantification of clinical signs at physical examination. J Intern Med 2003; 254: 257–263.

Purpose.  Although physical examination is a fundamental component of medical decision making, relatively few studies have evaluated how physicians quantify clinical signs and whether different methods of assessment have different effects on clinical practice.

Objectives.  To evaluate a possible impact of clinical experience when attending physicians, medical residents and medical students quantify qualitative signs of physical examination in a teaching hospital.

Setting.  Hospital das Clínicas, University of São Paulo, Brazil.

Subjects.  A total of 244 randomly selected physicians and medical students completed a reliable and consistent eight‐item questionnaire.

Main outcome measures.  To compare how they quantified clinical signs of cyanosis, anaemia, jaundice, oedema and dehydration, why they used the method(s) they described, and whether the method used could affect diagnosis, further testing or patient management. A chi‐square test was used to calculate differences between the groups.

Results.  Whilst the majority of those surveyed tended to use a four‐level evaluation for these clinical signs, attending staff physicians were more likely to employ two‐level evaluations than were residents or medical students. For all five signs, attending physicians’ use of dichotomous evaluations was significantly higher than that of residents or medical students: anaemia (P = 0.004), cyanosis (P < 0.001), oedema (P = 0.005), dehydration (P < 0.001) and jaundice (P = 0.002).

Conclusion.  Although medical students and residents are routinely taught to use a four‐level evaluation for these clinical signs, many of those surveyed tend to abandon this experience for a dichotomous approach. Given that the clinicians in this survey tended not to change their initial approach to a patient based on the intensity of this semi‐quantitative method, increased emphasis on teaching dichotomous approach evaluations in medical school should be encouraged.
Effect of metabolic control on homocysteine levels in type 2 diabetic patients: a 3‐year follow‐up
Authors: Passaro A. , Calzoni F. , Volpato S. , Nora E. Dalla , Pareschi P. L. , Zamboni P. F. , Fellin R. , Solini A.
Published: August 2003

Abstract.  Passaro A, Calzoni F, Volpato S, Dalla Nora E, Pareschi PL, Zamboni PF, Fellin R, Solini A (University of Ferrara; Diabetes Division Arcispedale S. Anna, Ferrara; and University of Pisa, Italy). Effect of metabolic control on homocysteine levels in type 2 diabetic patients: a 3‐year follow‐up. J Intern Med 2003; 254: 264–271.

Objectives.  Hyperhomocysteinaemia has emerged as a novel risk factor for cardiovascular disease. The determinants of total homocysteine (tHcy) levels in type 2 diabetic patients (D2p) have not been studied in detail. We examined prospectively the effect of different degrees of metabolic control on plasma tHcy in D2p with preserved kidney function.

Subjects and main outcome measurements.  Ninety‐five D2p were studied. Clinical parameters, fasting plasma glucose, HbA1c, serum lipids, blood urea nitrogen (BUN) and creatinine, vitamin B12 and folate and tHcy were measured at the baseline and after 36 months. The methylentetrahydrofolate reductase (MTHFR) C677T polymorphism was also determined. Subjects were categorized according to δHbA1c into group A (±1 point), B (>1 point increase) or C (>1 point decrease).

Results.  Total homocysteine was reduced in subjects whose HbA1c decreased with time, whilst patients showing a worsened metabolic control had an increased tHcy in respect to baseline. A larger response to the improved metabolic control in terms of tHcy reduction was noted in wild type patients versus those homozygous for the mutation. A multivariate analysis revealed MTHFR polymorphism and HbA1c as strong determinants of changes in tHcy with time.

Conclusions.  The findings suggest that in D2p tHcy decreases even with modest improvement of glycaemic control; moreover patients homozygous for the MTHFR C677T mutation show the largest changes in tHcy levels with concomitant changing of HbA1c. These results define a further mechanism through which hyperglycaemia might promote cardiovascular damage in diabetic patients.
A dietary and behavioural programme for the treatment of obesity. A 4‐year clinical trial and a long‐term posttreatment follow‐up
Authors: Lantz Helén , Peltonen Markku , Ågren Liselotte , Torgerson Jarl S
Published: August 2003

Abstract.  Lantz H, Peltonen M, Ågren L, Torgerson JS (Sahlgrenska University Hospital, Göteborg, Sweden). A dietary and behavioural programme for the treatment of obesity. A 4‐year clinical trial and a long‐term posttreatment follow‐up. J Intern Med 2003; 254: 272–279.

Objectives.  To evaluate weight loss maintenance after 4 years of nonpharmacological, nonsurgical obesity treatment, including a very low calorie diet (VLCD), diet and behavioural support. Furthermore, to assess weight development amongst completers and noncompleters beyond the active 4‐year treatment period.

Design.  Clinical trial.

Setting.  Two Swedish county hospitals.

Subjects.  A total of 113 patients were randomized to a 2‐year treatment programme with or without an initial VLCD period. The 87 patients who completed the 2‐year programme were offered the chance to continue a support programme for another 2 years. A total of 55 patients completed the entire 4‐year programme.

Interventions.  All the patients took part in a comprehensive support programme, including a hypocaloric diet and behavioural support, either as single treatment (non‐VLCD group) or following the VLCD period (VLCD group).

Results. Significant 4‐year weight losses were found in both groups, 7.6 ± 12.2 kg (VLCD group) and 6.3 ± 8.5 kg (non‐VLCD group), (P < 0.01, n.s. between groups). The completers (n = 55) had maintained a weight loss of 3.3 ± 10.7 kg (P < 0.05) 8 years after randomization. After 6 years, the noncompleters (n = 58) had gained 3.2 ± 9.7 kg compared with baseline (P < 0.05). The difference in weight change between completers and non‐completers was highly significant (P < 0.01).

Conclusions.  Highly significant weight losses can be maintained after a 4‐year comprehensive treatment programme, including a hypocaloric diet and behavioural support. An initial VLCD period did not significantly affect the long‐term weight loss. The posttreatment long‐term weight loss was larger amongst completers than amongst patients who did not complete the treatment.
Community‐acquired febrile urinary tract infection in diabetics could deserve a different management: a case–control study
Authors: Horcajada J. P. , Moreno I. , Velasco M. , Martínez J. A. , Moreno‐Martínez A. , Barranco M. , Vila J. , Mensa J.
Published: August 2003

Abstract.  Horcajada JP, Moreno I, Velasco M, Martínez JA, Moreno‐Martínez A, Barranco M, Vila J, Mensa J (Hospital Clínic Universitari‐IDIBAPS, Barcelona, Spain) Community‐acquired febrile urinary tract infection in diabetics could deserve a different management: a case–control study. J Intern Med 2003; 254: 280–286.

Objective.  To investigate if there are relevant differences in clinical, microbiological and outcome characteristics of community‐acquired febrile urinary tract infection (UTI) between diabetic and nondiabetic patients.

Design.  A prospectively matched case–control study.

Setting.  An 800‐bed tertiary care university‐affiliated hospital.

Subjects.  A total of 108 patients (54 diabetic and 54 nondiabetic patients matched by age and gender) admitted between January 1996 and September 1999 with febrile UTI.

Methods.  Clinical, analytical, microbiological and outcome variables were analysed by means of McNemar test (categorical) or Wilcoxon matched pairs signed rank test (continuous).

Results.  Mean age (SD) in both groups was 67.9 (14.4) years. In comparison with controls, diabetic patients were more likely to have fever without localizing symptoms (27% vs. 9%, P ≤ 0.0001), diminished consciousness level at admission (25% vs. 10%, P = 0.03), aetiological microorganism different from Escherichia coli (17% vs. 0, P = 0.0004), and quinolone‐resistant bacteria (17% vs. 3.7%, P = 0.07). Duration of fever after the onset of treatment was 1.75 (1) days in diabetics and 1.5 (1.1) days in nondiabetics (P = 0.17). However, diabetic patients had a longer hospitalization [5.2 (3.3) days] than nondiabetics [3.9 (2.6) days, P = 0.006].

Conclusions.  In diabetic patients, febrile UTIs have clinical and microbiological peculiarities that may have diagnostic and therapeutic implications.
Serum γ‐glutamyltransferase and development of impaired fasting glucose or type 2 diabetes in middle‐aged Japanese men
Authors: Nakanishi N. , Nishina K. , Li W. , Sato M. , Suzuki K. , Tatara K.
Published: August 2003

Abstract.  Nakanishi N, Nishina K, Li W, Sato M, Suzuki K, Tatara K (Osaka University Graduate School of Medicine, Osaka; and Japan Labor and Welfare Association, Tokyo; Japan). Serum γ‐glutamyltransferase and development of impaired fasting glucose or type 2 diabetes in middle‐aged Japanese men. J Intern Med 2003; 254: 287–295.

Objective.  To investigate the association between serum γ‐glutamyltransferase (GGT) and risk for development of diabetes.

Design.  Longitudinal study (followed from 1994 to 2001).

Setting.  A work site in Japan.

Subjects.  A total of 2918 Japanese male office workers aged 35–59 years who did not have impaired fasting glucose (IFG) (a fasting plasma glucose concentration of 6.1–6.9 mmol L−1), type 2 diabetes (a fasting plasma glucose concentration of 7.0 mmol L−1 or receipt of hypoglycaemic medication), medication for hypertension or hepatitis, alanine aminotransferase concentrations higher than three times the upper limit of the reference range or a history of cardiovascular disease at study entry.

Main outcome measure.  Incidence of IFG or type 2 diabetes over a 7‐year period.

Results.  With adjustment for potential risk factors for diabetes, the relative risk for IFG compared with serum GGT <16 U L−1 was 1.23 (95% CI, 0.79–1.90), 1.50 (CI, 0.97–2.32) and 1.70 (CI, 1.07–2.71) with serum GGT of 16–24, 25–43 and 44 U L−1, respectively (P for trend = 0.014). The respective relative risks for type 2 diabetes compared with serum GGT <16 U L−1 were 2.54 (CI, 1.29–5.01), 2.64 (CI, 1.33–5.23) and 3.44 (CI, 1.69–6.70) (P for trend = 0.002). From stratified analyses by body mass index (BMI) and alcohol intake, a stronger linear association between serum GGT and development of IFG or type 2 diabetes was found in men with a BMI 23.2 kg m−2 in both those who drank <46 and 46 g day−1 of ethanol.

Conclusions.  The risk for development of IFG or type 2 diabetes increased in a dose‐dependent manner as serum GGT increased in middle‐aged Japanese men. The increased relative risk for IFG or type 2 diabetes associated with serum GGT was more pronounced in obese men.
Causes and consequences of proteinuria: the kidney filtration barrier and progressive renal failure
Authors: Tryggvason K. , Pettersson E.
Published: August 2003

Abstract.  Tryggvason K, Pettersson E (Karolinska Institute, Stockholm, Sweden). Causes and consequences of proteinuria: the kidney filtration barrier and progressive renal failure (Review). J Intern Med 2003; 254: 216–224.

The past few years have witnessed a major breakthrough in the understanding of the molecular mechanisms and ultrastructural changes behind the development of proteinuria. The discovery of several proteins in the glomerular podocyte and slit diaphragm, where mutations lead to disease, has revealed the importance of this cell with its diaphragm as the major filtration barrier as opposed to the glomerular basement membrane (GBM) previously ascribed this function. Furthermore, accumulating clinical as well as experimental evidence points to the harmful effects of proteinuria, irrespective of the original damage. The purpose of this review is to shed light on what we know today about the two sides of this ‘coin’, the causes and the consequences of proteinuria.
Antibacterial peptides: basic facts and emerging concepts
Authors: Boman H. G.
Published: August 2003

Abstract.  Boman HG (Microbiology and Tumor Biology Center, Karolinska Institutet, Stockholm, Sweden). Antibacterial peptides: basic facts and emerging concepts (Review). J Intern Med 2003; 254: 197–215.

Antibacterial peptides are the effector molecules of innate immunity. Generally they contain 15–45 amino acid residues and the net charge is positive. The cecropin type of linear peptides without cysteine were found first in insects, whilst the defensin type with three disulphide bridges were found in rabbit granulocytes. Now a database stores more than 800 sequences of antibacterial peptides and proteins from the animal and plant kingdoms. Generally, each species has 15–40 peptides made from genes, which code for only one precursor. The dominating targets are bacterial membranes and the killing reaction must be faster than the growth rate of the bacteria. Some antibacterial peptides are clearly multifunctional and an attempt to predict this property from the hydrophobicity of all amino acid side chains are given. Gene structures and biosynthesis are known both in the fruit fly Drosophila and several mammals. Humans need two classes of defensins and the cathelicidin‐derived linear peptide LL‐37. Clinical cases show that deficiencies in these peptides give severe symptoms. Examples given are morbus Kostmann and atopic allergy. Several antibacterial peptides are being developed as drugs.
Population structure of North American beluga whales (Delphinapterus leucas) based on nuclear DNA microsatellite variation and contrasted with the population structure revealed by mitochondrial DNA variation
Authors: BROWN GLADDEN J. G. , FERGUSON M. M. , FRIESEN M. K. , CLAYTON J. W.
Published: August 2003

Beluga whales (Delphinapterus leucas) in North American waters migrate seasonally between wintering areas in broken pack ice and summering locations in estuaries and other open water areas in the Arctic and sub‐Arctic. Results from our previous investigation of beluga whale mitochondrial DNA (mtDNA) revealed genetic heterogeneity among beluga from different summering locations that was interpreted as representing a high degree of summering site philopatry. However, mtDNA is maternally inherited and does not reflect mating that may occur among beluga from different summering locations in wintering areas or during annual migrations. To test the possibility that breeding occurs among beluga from different summering locations, genetic variability at five nuclear DNA (nDNA) microsatellite loci was examined in the same animals tested in the mtDNA study. Beluga samples (n= 640) were collected between 1984 and 1994 from 24 sites across North America, mostly during the summer. Whales from the various sites were categorized into eight summering locations as identified by mtDNA analysis, as well as four hypothesized wintering areas: Bering Sea, Hudson Strait (Hudson Strait, Labrador Sea, southwest Davis Strait), Baffin Bay (North Water, east Davis Strait), and St Lawrence River. Microsatellite allele frequencies indicated genetic homogeneity among animals from summering sites believed to winter together but differentiation among whales from some of the wintering areas. In particular, beluga from western North America (Bering Sea) were clearly distinguished from beluga from eastern North America (Hudson Strait, Baffin Bay, and St Lawrence River). Based upon the combined data set, the population of North American beluga whales was divided into two evolutionarily significant units. However, the population may be further subdivided into management units to reflect distinct groups of beluga at summering locations.
Spread of violets in polluted pine forests: morphological and molecular evidence for the ecological importance of interspecific hybridization
Authors: NEUFFER B. , AUGE H. , MESCH H. , AMARELL U. , BRANDL R.
Published: August 2003

Hybridization between plant species occurs frequently but hybrids are often restricted to ecotones or disturbed habitats. In this study we show that introgressive hybrids between the tetraploidViola rivinianaand the diploidV. reichenbachianainvaded pine forests of the Dübener Heide (central Germany), an area affected by calcareous pollutants. The spread of these violet populations was correlated with the impact of pollution on habitat conditions. We compared morphology, cytology and random amplified polymorphic DNA (RAPD) bands among sixViolapopulations from the Dübener Heide and three populations of each pure species. RAPD analysis using 12 primers revealed 141 scorable bands. We considered bands as species specific if they occurred in at least 75% of the plants in one pure violet species but in none of the other. Seven bands were specific toV. rivinianaand 11 bands were specific toV. reichenbachiana. Two plants of aV. reichenbachianapopulation were identified as hybrids. Of theViolapopulations from the Dübener Heide, one was diploid and could be classified asV. reichenbachianaby morphology and RAPD markers. However, the majority of the Dübener Heide populations were tetraploid, and of a more variable morphology thanV. rivinianaandV. reichenbachiana, showing different combinations of intermediate characters, characters of the pure species and extreme characters. Despite their overall genetic similarity toV. riviniana, these plants could be identified as introgressive hybrids betweenV. rivinianaandV. reichenbachianaby species‐specific RAPD bands. Therefore, we propose that recurrent hybridization and backcrossing resulted in novel genotypes adapted to the changed environment of polluted pine forests.
A modular esterase from Penicillium funiculosum which releases ferulic acid from plant cell walls and binds crystalline cellulose contains a carbohydrate binding module
Authors: Kroon Paul A. , Williamson Gary , Fish Neville M. , Archer David B. , Belshaw Nigel J.
Published: August 2003

An esterase was isolated from cultures of the filamentous fungus Penicillium funiculosum grown on sugar beet pulp as the sole carbon source. The enzyme (ferulic acid esterase B, FAEB) was shown to be a cinnamoyl esterase (CE), efficiently releasing hydroxycinnamic acids from synthetic ester substrates and plant cell walls, and bound strongly to microcrystalline cellulose. A gene fragment was obtained by PCR using partial amino‐acid sequences obtained from the pure enzyme and used to a probe a P. funiculosum genomic DNA library. A clone containing a 1120‐bp ORF, faeB, was obtained which encoded a putative 353‐residue preprotein including an 18‐residue signal peptide, which when expressed in Eschericia coli produced CE activity. Northern analysis showed that transcription of faeB was tightly regulated, being stimulated by growth of the fungus on sugar beet pulp but inhibited by free glucose. The faeB promoter sequence contains putative motifs for binding an activator protein, XLNR, and a carbon catabolite repressor protein, CREA. FAEB was comprised of two distinct domains separated by a 20 residue Thr/Ser/Pro linker region. The N‐terminal domain comprised 276 amino acids, contained a G‐X‐S‐X‐G motif typical of serine esterases, and was shown to be a member of a family comprising serine esterases, including microbial acetyl xylan esterases, poly (3‐hydroxyalkanoate) depolymerases and CEs, and proteins of unknown function from Mycobacterium spp. and plants. The C‐terminal domain comprised 39 amino acids and closely resembled the family 1 cellulose binding carbohydrate‐binding modules (CBM) of fungal glycosyl hydrolases. This is the first report of a fungal CE with a CBM.
Interplay between repressing and activating domains defines the transcriptional activity of IRF‐1
Authors: Kirchhoff Sabine , Oumard André , Nourbakhsh Mahtab , Levi Ben‐Zion , Hauser Hansjörg
Published: August 2003

Interferon regulatory factor‐1 (IRF‐1) is a transcriptional activator with weak activation capacity. By defining the transcriptional activation domain of IRF‐1 we identified two activator fragments located between amino acids 185 and 256 functioning in an additive manner. Another fragment of IRF‐1, which has no activator function alone, acts as a strong enhancer element of these activator sequences. This enhancer element resides between the activator domains and the C‐terminus. In addition, we identified a novel type of inhibitory domain in the N‐terminal 60 amino acids of IRF‐1 which strongly inhibits its transcriptional activity. Because this fragment is conserved in all interferon regulatory factors, we found similar repression effects in the corresponding fragments in IRF‐2, IRF‐3 and interferon consensus sequence binding protein (ICSBP/IRF‐8). Interestingly, the corresponding sequence in p48/IRF‐9 is divergent, so that it does not show this inhibitory activity. A five‐amino‐acid sequence distinguishes the p48/IRF‐9 N‐terminus from the homologous parts in other interferon regulatory factors containing the repressing function. Replacing the diverged amino acids in IRF‐1 with the corresponding sequence of p48/IRF‐9 resulted in a loss of inhibitory activity within IRF‐1. The opposing activities within interferon regulatory factors may contribute to balanced or tuned regulation of gene activation, depending on the promoter context.
Nitration and inactivation of cytochrome P450BM‐3 by peroxynitrite
Authors: Daiber Andreas , Herold Susanna , Schöneich Christian , Namgaladze Dmitry , Peterson Julian A. , Ullrich Volker
Published: August 2003

Peroxynitrite (PN) is likely to be generated in vivo from nitric oxide and superoxide. We have previously shown that prostacyclin synthase, a heme‐thiolate enzyme essential for regulation of vascular tone, is nitrated and inactivated by submicromolar concentrations of PN [Zou, M.‐H. & Ullrich, V. (1996) FEBS Lett.382, 101–104] and we have studied the effect of heme proteins on the PN‐mediated nitration of phenolic compounds in model systems [Mehl, M., Daiber, A. & Ullrich, V. (1999) Nitric Oxide: Biol. Chem.2, 259–269]. In the present work we show that bolus additions of PN or PN‐generating systems, such as SIN‐1, can induce the nitration of P450BM‐3 (wild‐type and F87Y variant), for which we suggest an autocatalytic mechanism. HPLC and MS‐analysis revealed that the wild‐type protein is selectively nitrated at Y334, which was found at the entrance of a water channel connected to the active site iron center. In the F87Y variant, Y87, which is directly located at the active site, was nitrated in addition to Y334. According to Western blots stained with a nitrotyrosine antibody, this nitration started at 0.5 µm of PN and was half‐maximal between 100 and 150 µm of PN. Furthermore, PN caused inactivation of the P450BM‐3 monooxygenase as well as the reductase activity with an IC50 value of 2–3 µm. As two thiol residues/protein molecule were oxidized by PN and the inactivation was prevented by GSH or dithiothreitol, but not by uric acid (a powerful inhibitor of the nitration), our data strongly indicate that the inactivation is due to thiol oxidation at the reductase domain rather then to nitration of Y residues. Stopped‐flow data presented here support our previous hypothesis that ferryl‐species are involved as intermediates during the reactions of P450 enzymes with PN.
Mechanisms of differential transferrin receptor expression in normal hematopoiesis
Authors: Sposi Nadia M. , Cianetti Luciano , Tritarelli Elena , Pelosi Elvira , Militi Stefania , Barberi Tiziano , Gabbianelli Marco , Saulle Ernestina , Kühn Lukas , Peschle Cesare , Testa Ugo
Published: August 2003

We have investigated the expression of transferrin receptor (TfR) iron regulatory protein‐1 (IRP‐1) and iron regulatory protein‐2 (IRP‐2) in liquid suspension culture of purified hematopoietic progenitor cells (HPCs) induced by a growth factor stimulus to proliferation and unilineage differentiation/maturation through the erythroid, granulocytic, monocytic and megakaryocytic lineages.

In initial HPC differentiation, TfR expression is induced in both erythroid and granulopoietic cultures. In late HPC differentiation (i.e. starting from day 5 of culture) and then differentiated precursor maturation, the TfR gene is highly expressed in the erythroid lineage, whereas it is sharply downmodulated in the granulopoietic, monocytopoietic and megakaryocytic series. The elevated TfR expression in erythroid cells is: (a) mediated through a high rate of TfR gene transcription; (b) modulated by intracellular iron levels; (c) mediated by TfR mRNA stabilization through the iron regulatory protein (IRP), in that IRP‐1 activity is high in erythroid lineage as compared to the levels observed in other hemopoietic lineages; and (d) dependent on exogenous erythropoietin (Epo) (this is indicated by the marked TfR and IRP‐1/IRP‐2 downmodulation after Epo starvation).

Interestingly, analysis of IRP‐1 and IRP‐2 expression during hemopoietic differentiation showed that: (a) IRP‐1 expression was maintained during all steps of erythroid differentiation, while it was lost in the other hemopoietic lineages; (b) IRP‐2 expression was observed during all stages of hemopoietic differentiation in all four lineages. However, IRP‐1 and IRP‐2 expression and activity are induced when monocytes, which express only low levels of IRP‐1 and IRP‐2, are induced to maturation to macrophages.

These studies indicate that: (a) in normal erythropoiesis, the hyperexpression of TfR, starting from early erythroid HPC differentiation, is Epo‐dependent and mediated via transcriptional and post‐transcriptional mechanisms; (b) in the granulopoietic, monocytopoietic and megakaryocytic pathways, the TfR is first induced and then downmodulated (the latter phenomenon is mediated via transcriptional suppression of the TfR gene and IRP inactivation).
Isolation and characterization of recombinant antibody fragments against CDC2a from Arabidopsis thaliana
Authors: Eeckhout Dominique , Fiers Esbjörn , Sienaert Rebecca , Snoeck Veerle , Depicker Ann , De Jaeger Geert
Published: August 2003

In order to obtain recombinant antibody fragments that bind the cell‐cycle protein CDC2a from Arabidopsis thaliana (CDC2aAt), two phage display libraries of single‐chain variable (scFv) fragments were constructed. One library was derived from mice immunized with recombinant CDC2aAt N‐terminally fused to a His6‐tag (His‐CDC2aAt) and the other was made out of an anti‐PSTAIRE hybridoma cell line. Six specific His‐CDC2aAt‐binding phage clones (3D1, 3D2, 3D10, 3D25, 4D21 and 4D47) were isolated by panning. The isolated monoclonal phage clones, as well as the soluble scFv fragments produced in the periplasm of Escherichia coli, bind His‐CDC2aAt in ELISA and on Western blots. Moreover, four clones (3D1, 3D2, 3D10 and 4D21) detect specifically CDC2aAt from Arabidopsis cell suspensions on Western blots. Clone 4D21 binds the PSTAIRE epitope, whereas the 3D1, 3D2 and 3D10 clones bind, as yet unidentified, epitopes of CDC2aAt. Furthermore, the accumulation and antigen‐binding activity of these scFv fragments in a reducing environment were assessed. No interaction could be shown between the scFv fragments and CDC2aAt in a yeast two‐hybrid assay. However, after transient expression of the scFv fragments in the cytosol of tobacco leaves, three of six scFv fragments (3D1, 3D2 and 3D10) accumulated in the plant cytosol and ELISA results indicate that these scFv fragments retained antigen‐binding activity.
Conformational studies on a unique bis‐sulfated glycolipid using NMR spectroscopy and molecular dynamics simulations
Authors: Iida‐Tanaka Naoko , Fukase Koichi , Utsumi Hiroaki , Ishizuka Ineo
Published: August 2003

The time‐averaged solution conformation of a unique bis‐sulfated glycolipid (HSO3)2‐2,6Manα‐2Glcα‐1‐sn‐2,3‐O‐alkylglycerol, was studied in terms of the torsional angles of two glycosidic linkages, φ (H1‐C1‐O‐Cx) and ψ (C1‐O‐Cx‐Hx), derived from heteronuclear three‐bond coupling constants (3JC,H), and inter‐residual proton–proton distances from J‐HMBC 2D and ROESY experiments, respectively. The dihedral angles of Glcα1Gro in glycolipids were determined for the first time. The C1‐C4 diagonal line of the α‐glucose ring makes an angle of ≈ 120 ° with the glycerol backbone, suggesting that the α‐glucose ring is almost parallel to the membrane surface in contrast with the perpendicular orientation of the β‐isomer. Furthermore, minimum‐energy states around the conformation were estimated by Monte Carlo/stochastic dynamics (MCSD) mixed‐mode simulations and the energy minimization with assisted model building and energy refinement (AMBER) force field. The Glcα1Gro linkage has a single minimum‐energy structure. On the other hand, three conformers were observed for the Manα2Glc linkage. The flexibility of Manα2Glc was further confirmed by the absence of inter‐residual hydrogen bonds which were judged from the temperature coefficients of the chemical shifts, dδ/dT (−10−3 p.p.m.·°C−1), of hydroxy protons. The conformational flexibility may facilitate interaction of extracellular substances with both sulfate groups.
Muscarinic toxin‐like proteins from cobra venom
Authors: Kukhtina Viktoria V. , Weise Christoph , Muranova Tatyana A. , Starkov Vladislav G. , Franke Peter , Hucho Ferdinand , Wnendt Stephan , Gillen Clemens , Tsetlin Victor I. , Utkin Yuri N.
Published: August 2003

Three new polypeptides were isolated from the venom of the Thailand cobra Naja kaouthia and their amino‐acid sequences determined. They consist of 65‐amino‐acid residues and have four disulfide bridges. A comparison of the amino‐acid sequences of the new polypeptides with those of snake toxins shows that two of them (MTLP‐1 and MTLP‐2) share a high degree of similarity (55–74% sequence identity) with muscarinic toxins from the mamba. The third polypeptide (MTLP‐3) is similar to muscarinic toxins with respect to the position of cysteine residues and the size of the disulfide‐confined loops, but shows less similarity to these toxins (30–34% sequence identity). It is almost identical with a neurotoxin‐like protein from Bungarus multicinctus (TrEMBL accession number Q9W727), the sequence of which has been deduced from cloned cDNA only.

The binding affinities of the isolated muscarinic toxin‐like proteins towards the different muscarinic acetylcholine receptor (mAChR) subtypes (m1–m5) was determined in competition experiments with N‐[3H]methylscopolamine using membrane preparations from CHO‐K1 cells, which express these receptors. We found that MTLP‐1 competed weakly with radioactive ligand for binding to all mAChR subtypes. The most pronounced effect was observed for the m3 subtype; here an IC50 value of about 3 µm was determined. MTLP‐2 had no effect on ligand binding to any of the mAChR subtypes at concentrations up to 1 µm. MTLP‐1 showed no inhibitory effect on α‐cobratoxin binding to the nicotinic acetylcholine receptor from Torpedo californica at concentrations up to 20 µm.