bluuebunny/crossref_metadata_2025 huggingface dataset:
Intra‐ and inter‐laboratory variability in human T‐cell leukemia virus type‐1 proviral load quantification using real‐time polymerase chain reaction assays: A multi‐center study
Authors: Kamihira Shimeru , Yamano Yoshihisa , Iwanaga Masako , Sasaki Daisuke , Satake Masahiro , Okayama Akihiko , Umeki Kazumi , Kubota Ryuji , Izumo Shuji , Yamaguchi Kazunari , Watanabe Toshiki
Published: August 2010

Human T‐cell leukemia virus type‐1 (HTLV‐1) proviral load (VL) is an important determinant of viral pathogenesis and malignant evolution. Although VL has been quantified by in‐house real‐time quantifiable polymerase chain reaction (qPCR) technology, little is known about the harmonization among different VL assay systems. We evaluated intra‐ and inter‐laboratory variability of VL measured at six laboratories using the same DNA samples seropositive for HTLV‐1 in a two‐step manner. The first study measured 60 samples by original in‐house assays, finding that the median intra‐ and inter‐laboratory coefficient of variation (CV) was 44.9% (range, 25.4–71.8%) and 59.9% (34.2–93.4%), respectively. The inter‐laboratory correlation coefficients ranged from 0.760 to 0.875, indicating that VL were measured with good precision in each laboratory, but inter‐laboratory regression slopes differed from 0.399 to 2.206, indicating that VL were measured with a wide variation between laboratories. To examine the effect of standardization of reference materials (RM) on the VL variability, we performed a second study using only 20 samples by substituting RM for plasmid including the HTLV‐1 pX region. The median inter‐laboratory CV for raw pX copy number was reduced significantly from 66.9% to 35.7%, whereas the median CV for the internal control remained almost unchanged, resulting in no improvement in inter‐laboratory CV for VL. This indicates that each in‐house assay system worked well with good precision, but standardizing RM alone was insufficient for harmonization. The relevant choice of not only RM, but also internal control genes for data normalization is expected to be realistic to standardize HTLV‐1 VL measurement. (Cancer Sci 2010; 101: 2361–2367)
Clinical trial: once‐daily mesalamine granules for maintenance of remission of ulcerative colitis – a 6‐month placebo‐controlled trial
Authors: Lichtenstein G. R. , Gordon G. L. , Zakko S. , Murthy U. , Sedghi S. , Pruitt R. , Merchant K. , Shaw A. , Bortey E. , Forbes W. P.
Published: August 2010

Aliment Pharmacol Ther 2010; 32: 990–999

Background  Ulcerative colitis (UC) is a chronic relapsing and remitting idiopathic inflammatory bowel disorder.

Aim  To evaluate once‐daily mesalamine (mesalazine) granules (MG) for maintenance of remission of UC.

Methods  Randomized, double‐blind, placebo‐controlled trial of patients (n = 209 MG, n = 96 placebo) with UC in remission [revised Sutherland Disease Activity Index (SDAI) rectal bleeding = 0, mucosal appearance <2] who took MG 1.5 g or placebo once‐daily for up to 6 months. Primary efficacy endpoint: the percentage of patients who remained relapse‐free at month 6/end of treatment. Relapse was defined as SDAI rectal bleeding score ≥1 and a mucosal appearance score ≥2, a UC flare, or initiation of medication to treat a UC flare.

Results  The percentage of relapse‐free patients at month 6/end of treatment was higher with MG than placebo (78.9% vs. 58.3%, P < 0.001) in the intent‐to‐treat analysis. Significant differences (P ≤ 0.025) favouring MG were observed for most secondary endpoints including improvement in rectal bleeding, physician’s disease activity rating, stool frequency, the SDAI at month 6/end of treatment, patients classified as a treatment success and relapse‐free duration. The incidence of adverse events was similar between groups.

Conclusions  Once‐daily mesalamine (mesalazine) was effective in maintaining remission of UC for 6 months.
Increased diagnostic accuracy of atypical glandular cells in cervical liquid‐based cytology using cell blocks
Authors: Risse E. K. J. , Holierhoek J. P. , Meijer‐Marres E. M. , Ouwerkerk‐Noordam E. , Boon M. E.
Published: August 2010

E. K. J. Risse, J. P. Holierhoek, E. M. Meijer‐Marres, E. Ouwerkerk‐Noordam and M. E. Boon

Increased diagnostic accuracy of atypical glandular cells in cervical liquid‐based cytology using cell blocks

Objective:  The purpose of this study was to reduce the number of diagnoses of atypical glandular cells (AGC). Residual material from the cervical ThinPrep® samples (Hologic, Marlboruogh, MA, USA) was used for cell blocks (CB) and immunohistochemistry (IHC).

Methods:  In 2007 there were 87 patients (0.12% of tests) with AGC on liquid‐based cytology (LBC) in the Leiden Cytology and Pathology Laboratory (LCPL) using the Bethesda System 2001 (TBS). CB with IHC was used for 26 of these cases. The vials still containing the brush (Cervex‐Brush® Combi) were placed in a shaker for 10 minutes to dislodge the material trapped between the bristles. The residual sampling fluid was used to prepare paraffin sections (Shandon Cytoblock®) stained with Papanicolaou and immunostaining.

Results:  Four of five cases with AGC not otherwise specified (NOS) were diagnosed with CB/IHC as benign mimics (endometrium, tubal metaplasia, follicular cervicitis, microglandular hyperplasia) and one of four with AGC‐favour neoplasia (FN) (endocervical polyp). In one of five cases with AGC‐NOS and in two of seven with AGC‐FN, CIN3 was found on subsequent histological biopsy. Of six cases diagnosed as adenocarcinoma in situ (AIS) on LBC with CB/IHC the diagnosis was confirmed in four; one was adenocarcinoma and one glandular atypia. Of eight cases diagnosed as adenocarcinoma on cytology and CB/IHC, the diagnosis was confirmed in three. The other five cases were found to be one each of AIS, squamous cell carcinoma, CIN3, CIN2 with glandular atypia, and cervical endometriosis.

Conclusions:  By reducing the number of benign mimics of AGC, we achieved a high proportion (16/26; 61.5%) of neoplastic or preneoplastic lesions (glandular or squamous) on histological outcome potentially avoiding colposcopy. Histological biopsy verification by the gynaecologist is needed for final diagnosis of AGC‐FN, AIS and adenocarcinoma.
In situmeasurements confirm the seasonal dominance of benthic algae over phytoplankton in nearshore primary production of a large lake
Authors: MALKIN SAIRAH Y. , BOCANIOV SERGHEI A. , SMITH RALPH E. , GUILDFORD STEPHANIE J. , HECKY ROBERT E.
Published: August 2010

1. Despite the recognition of its importance, benthic primary production is seldom reported, especially for large lakes. We measuredin situbenthic net primary production by monitoring flux in dissolved inorganic carbon (DIC) concentration in benthic incubation chambers, based on continuous measurements of CO2(aq)flux, alkalinity, and the temperature‐dependent dissociation constants of carbonic acid (K1andK2). This methodology has the advantages of monitoring net primary production directly as change in carbon, maintaining continuous water recirculation, and having sufficient precision to detect change in DIC over short (i.e. 15 min) incubations, even in alkaline waters.

2. Benthic primary production onCladophora‐dominated rocky substrata in western Lake Ontario was measured biweekly. Maximum biomass‐specific net photosynthetic rates were highest in the spring (2.39 mgC g Dry Mass−1 h−1), decreased to negative rates by early summer (−0.76 mgC g DM−1 h−1), and exhibited a regrowth in late summer (1.98 mgC g DM−1 h−1).

3. ACladophoragrowth model (CGM), previously validated to predictCladophorabiomass accrual in Lake Ontario, successfully simulated the seasonality and magnitude of biomass‐specific primary production during the first cohort ofCladophoragrowth. Averaged over this growing season (May–Aug), mean areal net benthic production at the estimated depth of peak biomass (2 m) was 405 mg C m−2 d−1.

4. We measured planktonic primary production in proximity to the benthic study and constructed a depth‐resolved model of planktonic production. Using the CGM, benthic primary production was compared with planktonic primary production for the period May–Aug. Net benthic production from the shoreline to the 12 m contour (1–2 km offshore) equalled planktonic production. Closer to shore, benthic primary production exceeded planktonic primary production. Failure to account for benthic primary production, at least during abundantCladophoragrowth, will lead to large underestimates in carbon and nutrient flows in the nearshore zone of this Great Lake.
Complex controls on ostracod palaeoecology in a shallow coastal brackish‐water lake: implications for palaeosalinity reconstruction
Authors: HOLMES JONATHAN , SAYER CARL D. , LIPTROT ELLIE , HOARE DANIEL J.
Published: August 2010

1. The low‐Mg calcite shells of Ostracoda (Crustacea) are often well preserved in the sediments of alkaline lakes. In coastal waters that have undergone large temporal changes in salinity, ostracod assemblages preserved in the sediment record have been used to reconstruct palaeosalinity, often assuming that salinity is the only significant control on the faunas.

2. We evaluate the performance of ostracods as palaeosalinity indicators in Hickling Broad, a shallow brackish coastal lake in Norfolk, U.K., by comparing fossil ostracod assemblages covering two centuries with geochemical inferences and instrumental records of past salinity and water composition along with other palaeolimnological indicators.

3. Despite large changes in the salinity of the lake and the supposed salinity sensitivity of ostracods, the fossil ostracod assemblages do not clearly reflect the salinity trends inferred from the other independent data. Rather, a complex series of changes has occurred in the lake over the past 200 years and factors other than salinity, including eutrophication, toxicity and associated complex alterations in habitat availability have probably influenced ostracod assemblages. In contrast, there is a good broad agreement between inferred or measured salinity and the trace‐element chemistry of ostracod shells.

4. We conclude that ostracod faunas may not always provide unambiguous palaeosalinity records and should therefore not be used to reconstruct salinity changes except as part of a multi‐proxy investigation that includes other palaeoecological and/or geochemical indicators.
Disentangling stocking introgression and natural migration in brown trout: survival success and recruitment failure in populations with semi‐supportive breeding
Authors: WOLLEBÆK JENS , HEGGENES JAN , RØED KNUT H
Published: August 2010

1. Introgression into natural salmonid populations from stocked conspecifics has been widely studied. Outcomes vary from no effect even after decades of stocking, to population replacement after only a couple of generations. Potential introgression caused by semi‐supportive breeding (i.e. using a mixture of local strains as brood stock) is, however, less well studied.

2. We investigated population structure of brown trout (Salmo trutta) in a regulated alpine lake with three natural, environmentally contrasting tributaries used as spawning and rearing habitat. Massive semi‐supportive breeding of admixed local strains has been implemented for decades. Stocked trout represented c. 17% of the total lake population, and a substantial post‐release survival reflects a considerable potential for introgression. However, the mark‐recapture studies indicate no spawning runs of stocked fish.

3. Using 13 polymorphic microsatellite loci, we found natural straying and non‐native reproduction, especially among wild populations inhabiting environmentally unstable habitat. Retained genetic structure across tributaries indicated low reproductive success of wild‐born non‐natives. Moreover, the genetic structure among tributaries has probably not been influenced by semi‐supportive breeding, because of recruitment failure of stocked trout.
Chemical, physical and mechanical properties of a novel calcium aluminate endodontic cement
Authors: Oliveira I. R. , Pandolfelli V. C. , Jacobovitz M.
Published: August 2010

Oliveira IR, Pandolfelli VC, Jacobovitz M. Chemical, physical and mechanical properties of a novel calcium aluminate endodontic cement. International Endodontic Journal.

Aim  To evaluate the influence of additives on several physical and chemical properties of a novel endodontic cement based on calcium aluminate in comparison with mineral trioxide aggregate (MTA).

Methodology  Manipulation tests, measurements of temperature, pH, ionic conductivity, compressive strength, apparent porosity, and pore size distribution were carried out on Gray‐MTA (Angelus) and calcium aluminate cement (Secar‐71, Kerneos) with and without various additives: a polymeric dispersant, CaCl2 as plasticizer, and ZnO as radiopacifier.

Results  The calcium aluminate cement without additives had a setting time of approximately 60 min, and when combined with Li2CO3 it decreased to 10 min. The material also released Ca2+ ions and alkalinized the medium. Moreover, the addition of additives (dispersant, plasticizer, and radiopacifier) improved its properties resulting in a material with a viscosity of 57 mPa s, enhanced handling properties, a mechanical strength of 81 MPa, a porosity of 4% and pores with small diameter (0.25 μm). MTA had no temperature increase (that indicates setting) up to 400 min, a mechanical strength of 34 MPa, and porosity of 28% with pores 2.5 μm in diameter.

Conclusion  The novel cement set more rapidly, had better fluidity, improved handling properties, higher mechanical strength, and reduced porosity with lower pore size compared to Gray‐MTA Angelus.
Is a more active attitude warranted in patients with subclinical thyrotoxicosis?
Authors: TENERZ Å. , FORBERG R. , JANSSON R.
Published: August 2010

Abstract. In 1985 one of the new sensitive thyroid stimulating hormone (TSH) assays was introduced as part of our laboratory routine for thyroid function testing. Consequently, we now routinely identify a small but not insignificant group of patients with ‘subclinical thyrotoxicosis’, i.e. a low serum TSH in conjunction with a normal serum free T4. We here present the results of a 2‐year follow‐up investigation, which includes 40 patients with subclinical thyrotoxicosis and 40 euthyroid control patients. The group with subclinical thyrotoxicosis was characterized by a mean age of 65 years and a high prevalence of nodular goitre. Twelve (30%) of the patients but none of the individuals in the control group were treated during the follow‐up period because of clinical thyroid disease. Atrial fibrillation was found in 11 (28%) patients compared to four (10%) of the controls. Therapy should be considered more often than previously in patients with nodular goitre and subclinical thyrotoxicosis, particularly in conjunction with atrial fibrillation.
Associations between pituitary–adrenocortical function and abdominal obesity, hyperinsulinaemia and dyslipidaemia in normotensive males
Authors: HAUTANEN A. , RÄIKKÖNEN K. , CREUTZ H. ADLER
Published: August 2010

Abstract. Hautanen A, Räikkönen K, Adlercreutz H. (University of Helsinki, Helsinki, Finland) Associations between pituitary‐adrenocortical function and abdominal obesity, hyperinsulinaemia and dyslipidaemia in normotensive males.

Objectives: To examine the relationships between pituitary‐adrenal cortical (PA) function, abdominal obesity, hyperinsulinaemia, and dyslipidaemia.

Design: A prospective study.

Setting: Helsinki University Central Hospital, Finland.

Subjects: Seventy‐one healthy males aged 30–55 years.

Main outcome measures: Insulin sensitivity was assessed by the oral glucose tolerance test (OGTT). Basal PA activity was examined by measuring urinary and serum concentrations of hormones, followed by dexamethasone suppression and corticotrophin (ACTH) stimulation tests to determine functional PA activity.

Results: The means of waist‐to‐hip ratio (WHR), body‐mass index (BMI), HDL‐cholesterol and triglyceride levels, and insulin and C‐peptide measurements during the OGTT were significantly different across the tertiles for insulin: glucose ratio. The ratio of 12‐h urinary cortisol excretion to BMI, preceding the OGTT, and the mean basal cortisol level during the OGTT were decreased, while the net increments of cortisol and 17‐hydroxyprogesterone (17‐OHP) from 0 to 60 min, as well as the ratio of net 17‐OHP to 11‐deoxycortisol increments, after ACTH, were elevated in the upper compared with the lower tertile. The mean cortisol during the OGTT, and the ratio of urinary cortisol to BMI were negatively related, while absolute DHEA and cortisol responses to ACTH were positively related to fasting and mean insulin levels. Hormonal variables, WHR. insulin, and triglycerides were successfully integrated into a tentative mathematical model by the use of covariance structure (path) analyses.

Conclusions: Several alterations in the PA function, suggestive of decreased 21‐hydroxylase activity, mild cortisol deficiency and slight adrenal hyperplasia, are associated with abdominal obesity which, in turn, appears to be an important prelude to insulin resistance and dyslipidaemia.
Cancer and diabetes ‐ a follow‐up study of two populationbased cohorts of diabetic patients
Authors: HJALGRIM H. , FRISCH M. , EKBOM A. , KYVIK K. O. , MELBYE M. , GREEN A.
Published: August 2010

Abstract. Hjalgrim H, Frisch M, Ekbom A, Kyvik KO, Melbye M, Green A (Statens Serum Institut, Copenhagen; Odense University, Odense; Denmark; and University Hospital, Uppsala, Sweden). Cancer and diabetes ‐ a follow‐up study of two population‐ based cohorts of diabetic patients.

Objectives: To study the occurrence of cancer amongst patients with diabetes mellitus (DM).

Design: Population based cohort study.

Setting: Denmark.

Subjects: Two cohorts of patients with DM were identifled. One cohort comprised 1659 conscripts diagnosed with type I DM before the age of 20 years. Another cohort comprised 1499 men and women with insulin treated DM identified by means of medical prescriptions on 1 July 1973. Both cohorts were followed until the end of 1992. Main outcome measures. The relative risk of cancer in the two cohorts was estimated as the ratio of observed to expected number of cancers in the cohort (SIR).

Results: No unusual risk of cancer was observed amongst the conscripts (SIR 0.9, n = 13) or amongst patients with onset of DM before the age of 30 years in the prescription cohort (SIR 0.9, n = 32). Amongst those aged 30 years or more at DM onset in the prescription cohort, the overall risk of cancer did not depart from normal (SIR 1.0, n = 103), however, pancreatic cancer occurred in excess both immediately (< 1 year) (SIR 190, n = l), and during 1–9 years after DM onset (SIR 9.0, n = 4). Similarly, the risk of non‐Hodgkin's lymphoma was increased significantly (SIR 3.3, n = 6), all cases occurring more than 10 years after DM onset.

Conclusions: Our data suggest that there is no unusual risk of cancer associated with type I DM. Type II DM may be the first symptom of pancreatic cancer and may be associated with an increased risk of non‐Hodgkin's lymphoma.
Association of BsmI vitamin‐D receptor gene polymorphism with combined bone mass in spine and proximal femur in Icelandic women
Authors: SIGURDSSON G. , MAGNUSDOTTIR D. N. , KRISTINSSON J. Ö. , KRISTJANSSON K. , OLAFSSON I.
Published: August 2010

Abstract. Sigurdsson G, Magnusdottir DN, Kristiisson J0, Kristjansson K, Olafsson I (Reykjavik Hospital, University of Iceland, Reyjavik, Iceland). Association of BsmI vitamin‐D receptor gene polymorphism with combined bone mass in spine and proximal femur in Icelandic women.

Objective: To evaluate whether there is an association between BsmI‐vitamin‐D receptor (VDR) gene polymorphism and combined bone mass in the spine and proximal femur in a group of adult Icelandic women with high and low bone mineral density (BMD).

Design: Comparison of distribution of VDR genotypes (BB, Bb and bb) and allele frequency (B and b) in two groups of women: a group with ‘strong bones’ with high BMD in both the spine and proximal femur (< 1 standard deviation [SD]) above the age‐matched mean (n = 3 5) and a group with ‘weak bones’ with BMD < 1.5 SD below the age‐matched mean at both sites using dual energy X‐ray absorptiometry.

Setting: Iceland, a population with a mean calcium intake > 1000 mg day‐'. The calcium intake in the study group was however not evaluated.

Subjects: Eighty‐three Icelandic women, aged 22–65, free of diseases affecting bone and not taking drugs affecting calcium or bone metabolism, recruited from women undergoing bone densitometry at the Reykjavik Hospital.

Main outcome measures: Frequency of VDR genotypes and alleles in the two groups.

Results: The distribution of VDR genotypes was women with high and low bone mineral density significantly different in the two groups (P < 0.01): the b allele frequency was 70% in the group with high BMD compared to 48.5 % in the group with low BMD.

Conclusions: In this selected group of adult Icelandic women the b allele in the vitamin‐D receptor gene seems to be associated with high bone mass in the spine and proximal femur.
Thyrotoxic period paralysis: report of three cases and review of the literature.
Authors: PAPADOPOULUS K. I. , DIEP T. , CLELAND B. , LUNN N. W.
Published: August 2010

Abstract. Papadopoulos KI, Diep T, Cleland B. Lunn N W (Fairfield Hospital, Wetherill Park, Australia) Thyrotoxic periodic paralysis: report of three cases and review of the literature (Case report.

Thyrotoxic periodic paralysis (TPP) is a neuromuscular disoder that, in a global perspective, only affects a small percentage of patients with thyrotoxicosis of any aetiology, but is a fairly common phenomenon in Asian populations where it predominantly ouucrs in males. Three typical cases are presented here. All three patients were Asian males, one being the first Laotian patient reported to have this disorder. In one case prodromal symptoms of TPP occurred during a relapse of the thyrotoxicosis. The epidemiology, clinical manifestations, aetiology and treatment of this relatively rare condition are discussed. As oriental races are now frequently ecnountered in the Western world, it is imperative that TPP is included in the differential diagnosis of muscle weakness in order to avoid the life‐threatening complications of hypokalaemia and thyrotoxicosis.
Cisapride increases peak plasma and saliva ethanol levels under fasting conditions
Authors: DZIEKAN G. , CONTESSE J. , WERTH B. , SCHWARZER G. , REINHART W. H.
Published: August 2010

Abstract. Dziekan G, Contesse J, Werth B, Schwarzer G. Keinhart WH (Kantonsspital, Chur, Switzerland and University of Freiburg, Freiburg, Germany). Cisapride increases peak plasma and saliva ethanol levels under fasting conditions. J Intern Med 1997; 242: 479‐482.

Objectives: Alcohol absorption is influenced by gastric first‐pass metabolism through an alcohol dehydrogenase and the gastric emptying time. Whilst an influence of antisecretory drugs and aspirin on gastric alcohol metabolism has been described, the role of prokinetic drugs has not been determined.

Design: A randomized, placebo‐controlled doubleblind cross‐over study was performed.

Setting: Out‐patient facilities of a referral hospital.

Subjects: Eight male volunteers (age range 25–46 years).

Intervention: Treatment with two doses of either placebo or cisapride 150 pgkg 7 h and 20 min before drinking 0.5 g/kg alcohol either in a fasting state or during a standardized meal (12 kcalkg).Main outcome measures. Plasma and saliva ethanol concentrations were measured during 4 h. Results. Cisapride increased peak plasma ethanol levels in fasting subjects from 15.6 (SD 1.4, 95′%‐KI 14.7i16.6) to 17.8 (SD 2.7, 9SYo‐KI 15.9i19.7)mmol/L and saliva ethanol 30 min after alcohol ingestion from 11.4 (SD 2.2, 95%‐KI 9.9;12.9) to 15.9 (SD 4.3, 95Yo‐KI 12.9;18.8) mmol/L. A significant interaction between fasting state and drug intake was found for the 30 min saliva ethanol values (P < 0.05, ANOVA for repeated measurements).

Conclusions: Cisapride may increase ethanol levels under fasting conditions. Patients treated with prokinetic drugs should be informed about this possibility.
Colitis as the initital presentation of Wegener's granulomatosis.
Authors: SCHNEIDER A. , MENZEL J. , GAUBITZ M. , KELLER R. , LÜGERING N. , DOMSCHKE W.
Published: August 2010

Abstract. Schneider A, Menzel J, Gaubitz M, Keller R, Domschke W (University of Munster, Munster, Germany). Colitis as the initial presentation of Wegener's granulomatosis (Case Report).

Wegener's granulomatosis is a vasculitis which commonly presents with symptoms referable to the upper airway system, lungs, joints, eyes and kidneys. We present a case in which colitis combined with sinusitis could retrospectively be attributed as the initial manifestations of Wegener's disease.
What else adds to hypertension in predicting stroke? An incident case–referent study
Authors: Öhgren B. , Weinehall L. , Stegmayr B. , Boman K. , Hallmans G. , Wall S.
Published: August 2010

Abstract. Öhgren B, Weinehall L, Stegmayr B, Boman K, Hallmans G, Wall S (Umeå University and Skellefteå County Hospital, Umeå, Sweden). What else adds to hypertension in predicting stroke? An incident case–referent study. J Intern Med 2000; 248: 475–482.

Objectives. To examine to what extent traditional biomedical risk factors and social factors can predict, separately or jointly, first‐ever stroke event, and to explore to what extent other risk factors might add to hypertension/elevated blood pressure as a stroke risk factor.

Design. An incident case–referent study.

Setting. The study was nested within the Västerbotten Intervention Program (VIP) and the Northern Sweden MONICA cohorts.

Subjects. The study involves 129 stroke cases with two randomly selected referents per case from the same study cohorts.

Results. History of diabetes, daily smoking, obesity (body mass index ≥ 30 kg m−2), hypertension and living as an unmarried person were associated with an increased risk in univariate analysis. However, in the multivariate logistic regression analysis, only hypertension, diabetes and single marital status remained significant. There was a statistically significant interaction between hypertension and smoking and stroke. Interaction of possible clinical relevance was also observed between hypertension, history of diabetes and stroke.

Conclusions. The study both emphasizes the multifactorial nature of stroke and illustrates that knowledge of interactions offers different strategic approaches when preventing stroke in clinical work. It is important, but not enough, to focus on blood pressure control. Diabetes prevention as well as prevention of smoking must also be considered as factors of major importance.
Molecular epidemiology
Authors: Boffetta P.
Published: August 2010

Abstract. Boffetta P (Unit of Environmental Cancer Epidemiology, International Agency for Research on Cancer, Lyon, France). Molecular epidemiology (Internal Medicine in the 21st Century). J Intern Med 2000; 248: 447–454.

The use of biomarkers in epidemiology is not new, but recent developments in molecular biology and genetics have increased the opportunities for their use. However, epidemiological studies based on biomarkers, which belong to the discipline defined as ‘molecular epidemiology’ are subject to the same problems of design and analysis as ‘traditional’ epidemiological studies. If biomarkers offer new opportunities to overcome some of the limitations of epidemiology, their added value over traditional approaches should be systematically assessed. Biomarkers should be validated though transitional studies; consideration to sources of bias and confounding in molecular epidemiology studies should be no less stringent than in traditional studies.
Hormone replacement therapy in healthy postmenopausal women: a randomized, placebo‐controlled study of effects on coagulation and fibrinolytic factors
Authors: Gottsäter A. , Rendell M. , Hulthén U. L. , Berntorp E. , Mattiasson I.
Published: August 2010

Abstract. Gottsäter A, Rendell M, Hulthén UL, Berntorp E, Mattiasson I (University of Lund, University Hospital, Malmö, Sweden). Hormone replacement therapy in healthy postmenopausal women: a randomized, placebo‐controlled study of effects on coagulation and fibrinolytic factors. J Intern Med 2001; 249: 237–246.

Objectives. To evaluate effects of postmenopausal hormone replacement therapy (HRT) on von Willebrand factor, factor (F)VIII, factor (F)VII, fibrinogen, antithrombin (AT) III, prothrombin fragments 1 and 2, protein C, total and free protein S, plasminogen activator inhibitor‐1 (PAI‐1), tissue plasminogen activator (tPA) and resistance to activated protein C.

Design. Part 1: double blind randomized trial for 3 months. Part 2: open study for 9 months.

Setting. Department of Endocrinology, University Hospital, Malmö, Sweden.

Subjects. Fifty‐one postmenopausal women with a history of amenorrhoea of at least 6 months and body mass index ≥ 24 kg m−2 participated in part 1 and 46 participated in part 2.

Intervention. Randomization for placebo (n=24) or HRT (n=27). HRT was given as 2 mg oestradiol valerate for the first 3 months, with the addition of 10 mg medroxyprogesterone for 10 days every third month thereafter.

Measurements. At baseline and after 3 and 12 months.

Results. During 0–3 months in the HRT group, FVII increased (P < 0.01), whereas fibrinogen, AT III and total protein S all decreased (P < 0.001 for all). Changes in variables were expressed as Δ‐values. After 3 months Δ‐values differed between groups for fibrinogen (P < 0.05), AT III (P < 0.001), total protein S (P < 0.001), and PAI‐1 (P < 0.001). During 0–12 months, fibrinogen, total protein S, tPA (P < 0.01 for all) and AT III (P < 0.05) decreased. In the control group, all variables were unchanged during the study, except for increases (P < 0.05) in total protein S after 3 and 12 months, and a decrease (P < 0.01) in FVIII after 12 months. After 12 months Δ‐values differed for fibrinogen (P < 0.05), AT III (P < 0.05) and total protein S (P < 0.001).

Conclusions. Unopposed oestrogen substitution was associated with both potentially beneficial effects, such as decreases in fibrinogen, and potentially thrombogenic effects such as decreasing AT III and protein S and increasing FVII. During prolonged follow‐up and addition of progesterone, differences between groups concerning FVII were attenuated. These data suggest that effects of HRT upon coagulation are most pronounced early after institution of unopposed treatment.
Gender specific associations of the Trp64Arg mutation in the β3‐adrenergic receptor gene with obesity‐related phenotypes in a Mediterranean population: interaction with a common lipoprotein lipase gene variation
Authors: Corella D. , Guillén M. , Portolés O. , Sorlí J. V. , Alonso V. , Folch J. , Sáiz C.
Published: August 2010

Abstract. Corella D, Guillén M, Portolés O, Sorlí JV, Alonso V, Folch J, Sáiz C (School of Medicine, University of Valencia, Valencia, Spain). Gender specific associations of the Trp64Arg mutation in the β3‐adrenergic receptor gene with obesity‐related phenotypes in a Mediterranean population: interaction with a common lipoprotein lipase gene variation. J Intern Med 2001; 250: 348–360.

Objective. To investigate the association between the Trp64Arg β3‐adrenergic receptor (ADRB3) mutation and obesity‐related phenotypes in a Mediterranean Spanish population considering the effect of other genetic and environmental factors.

Design and subject. Cross‐sectional study in 1063 (476 men and 587 women) randomly selected from this population (aged: 18–68 years).

Measurements. Anthropometric (weight, height and waist‐to‐hip ratio), blood pressure, biochemical (lipids, fasting glucose, and uric acid), life‐style variables, and the Trp64Arg, HindIII‐Lipoprotein lipase (LPL) and apolipoprotein E polymorphism.

Results. Frequency of the Arg64 allele was low (0.051; 95% CI: 0.042–0.060). We found gender‐specific associations between the Trp64Arg mutation and obesity related phenotypes. In men, carriers of the Arg64 variant had higher body mass index (BMI) (27.63 ± 3.81 vs. 26.34 ± 3.57 kg m−2, P=0.049) and total cholesterol (5.85 ± 1.45 vs. 5.28 ± 1.06 mmol L−1; P=0.011) compared with wild‐type individuals. Logistic regression analysis, revealed that the risk of overweight was two times higher in male carriers of the Arg64 allele. In women, the Arg64 variant was only associated with higher fasting glucose (P=0.031). These genotype effects persisted after adjustment for age, genetic and life‐style variables. For the LPL polymorphism, the H−/H− genotype was associated with lower BMI and with lower risk of overweight (OR: 0.49; 95% CI: 0.30−0.81) in both men and women. However, after adjustment for covariates, these associations only remained statistically significant (P < 0.02) in women. Moreover, in women, a statistically significant interaction (P=0.026) between the LPL and the ADRB3 gene loci in determining BMI was found. Thus, the Arg64 allele was associated with a higher BMI only in H+/H+ women.

Conclusions. The Trp64Arg mutation was associated with BMI and lipids in men. In women, an additional gene–gene interaction with the LPL‐HindIII polymorphism may explain the results.
Tumour staging, morphology and p53 overexpression concur in predicting survival in hepatocellular carcinoma
Authors: GIANNI S. , CECCHETTO A. , ALTAVILLA G. , RAGAZZI R. , BERTAZZO M. , DE GIORGIO M. , BALDAN A. , FAGIUOLI S. , FARINATI F.
Published: August 2010

Background/aims.  The prognosis of hepatocellular carcinoma (HCC) on cirrhosis is hard to predict as it depends on tumour stage, underlying liver disease, type of treatment and, possibly, biological factors of the tumour itself.

Methods.  We prospectively evaluated the survival of 91 consecutive patients with HCC on cirrhosis, diagnosed between January 1998 and December 1999. Clinical features and histological/biological aspects, including histotype, grade, p53 overexpression, cytoproliferation and apoptotic markers were analysed.

Results.  Child‐Pugh (P = 0.01), Okuda (P < 0.0001), Cancer of the Liver Italian Program (CLIP) staging (P < 0.0001) and type of treatment (P = 0.0001) were significantly related to survival. In the Cox model, CLIP staging was included as independent predictor of survival at step 1 (P < 0.0001) with Okuda at step 2 (P = 0.013). Amongst the biological factors, p53 overexpression and histotype were significantly related with survival (P = 0.0044 and 0.017 respectively). When clinical and biological variables were examined together in the Cox model, CLIP and Okuda were confirmed as being statistically related with survival (P < 0.0001 and =0.012) followed by histotype and p53 overexpression (P = 0.019 and 0.02).

Conclusions.  CLIP, Okuda, histotype and p53 overexpression are the strongest predictors of survival in this series of patients. These data confirm that staging of the tumour and underlying liver disease are strictly related to prognosis but support the concurrent role of clinical and biological factors in upgrading our capacity of predicting the fate of HCC patients.
Zoonotic transmission of Cryptococcus neoformans from a magpie to an immunocompetent patient
Authors: LAGROU K. , VAN ELDERE J. , KEULEERS S. , HAGEN F. , MERCKX R. , VERHAEGEN J. , PEETERMANS W. E. , BOEKHOUT T.
Published: August 2010

Abstract.  Lagrou K, Van Eldere J, Keuleers S, Hagen F, Merckx R, Verhaegen J, Peetermans WE, Boekhout T (University Hospital Leuven, Leuven, Belgium; and Centraalbureau voor Schimmelcultures, CT Utrecht, the Netherlands) Zoonotic transmission of Cryptococcus neoformans from a magpie to an immunocompetent patient (Case report). J Intern Med 2005; 257: 385–388.

Abstract. 
We report a case of cryptococcal meningitis in an immunocompetent female patient with exposure to a pet magpie (Pica pica). Genetically indistinguishable isolates were cultured from the cerebrospinal fluid of the patient and excreta of the bird. Our data strongly suggest zoonotic transmission of Cryptococcus neoformans var. grubii from a magpie to this patient.
Fracture risk associated with systemic and topical corticosteroids
Authors: VESTERGAARD P. , REJNMARK L. , MOSEKILDE L.
Published: August 2010

Objectives.  To study the fracture risk associated with the use of corticosteroids in any formulation and administration.

Design.  Case–control study.

Setting.  Community‐based study in Denmark.

Subjects.  Cases were all subjects with any fracture sustained during the year 2000 (n = 124 655). For each case, three controls (n = 373 962) matched on age and gender were randomly drawn from the background population. Adjustments were made for concurrent diseases (lung diseases, rheumatic disorders), use of other drugs (inhaled bronchodilators), contacts to hospitals and general practitioners, and social variables.

Results.  An increased risk of any fracture, hip, spine, and forearm fractures was present with use of more than 2.5 mg of prednisolone or equivalent orally per day. For inhaled corticosteroids a limited increase in the risk of any fracture was present in users of more than 7.5 mg prednisolone equivalents per day (equivalent to 1875 μg of budesonide per day). However, no increase in the risk of hip, spine or forearm fractures was present in users of inhaled corticosteroids. For other topical corticosteroids (dermal, rectal, nasal, local application in the mouth, the eyes or the ears) no increase in fracture risk could be demonstrated even at high doses after adjustment for confounders.

Conclusions.  Ingestion of more than 2.5 mg of oral prednisolone equivalents per day is associated with an increase in fracture risk. No increase is associated with inhaled corticosteroids except at daily dosages above 7.5 mg of prednisolone equivalents. No increase in fracture risk is associated with other forms of topical corticosteroids.
Neutrophil serine proteases: potential key regulators of cell signalling during inflammation
Authors: WIEDOW O. , MEYER‐HOFFERT U.
Published: August 2010

Abstract.  Wiedow O, Meyer‐Hoffert U (University Kiel, Kiel, Germany; and Karolinska Institute, Stockholm, Sweden). Neutrophil serine proteases: potential key regulators of cell signalling during inflammation (Review). J Intern Med 2005; 257: 319–328.

The serine proteases cathepsin G, human leucocyte elastase and proteinase 3 are major contents of neutrophils and are released at sites of inflammation. The common picture of their function was that they do not degrade extracellular proteins specifically. Recent studies provided evidence that these proteases are able to activate specifically pro‐inflammatory cytokines and lead to the activation of different receptors. Neutrophil serine proteases might therefore be important regulators of inflammatory processes and are interesting targets for new therapeutic approaches against inflammatory disorders. This review summarizes the current knowledge on the regulation of cell signalling by neutrophil serine proteases.
Separating cause from effect: how does insulin/IGF signalling control lifespan in worms, flies and mice?
Authors: Piper M. D. W. , Selman C. , McElwee J. J. , Partridge L.
Published: August 2010

Ageing research has been revolutionized by the use of model organisms to discover genetic alterations that can extend lifespan. In the last 5 years alone, it has become apparent that single gene mutations in the insulin and insulin‐like growth‐factor signalling pathways can lengthen lifespan in worms, flies and mice, implying evolutionary conservation of mechanisms. Importantly, this research has also shown that these mutations can keep the animals healthy and disease‐free for longer and can alleviate specific ageing‐related pathologies. These findings are striking in view of the negative effects that disruption of these signalling pathways can also produce. Here, we summarize the body of work that has lead to these discoveries and point out areas of interest for future work in characterizing the genetic, molecular and biochemical details of the mechanisms to achieving a longer and healthier life.
SecY and integral membrane components of the Escherichia coli protein translocation system
Authors: Ito Koreaki
Published: August 2010

Genetic approaches can address the question of how integral membrane Sec factors interact with each other and facilitate protein translocation across the cytoplasmic membrane of E. coli. This review summarizes genetic analyses of SecY, SecE and some other protein translocation factors, utilizing ‘prl’ mutations,‘sec’ mutations, ‘suppressor‐directed inactivation’, ‘Sec titration’, dominant negative mutations and their suppressors. Evidence suggests that coordinate participation of SecY, SecE, SecD, SecF, and probably some other factors, is crucial for the process.
Evidence for global regulatory control of pilus expression in Escherichia coli by Lrp and DNA methylation: model building based on analysis of pap
Authors: van der Woude M. W. , Braaten B. A. , Low D. A.
Published: August 2010

Pyelonephritis‐associated pilus (Pap) expression is regulated by a phase variation control mechanism involving PapB, Papl, catabolite activator protein (CAP), leucine‐responsive regulatory protein (Lrp) and deoxyadenosine methylase (Dam). Lrp and Papl bind to a specific non‐methylated pap regulatory DNA region containing the sequence ‘GATC’ and facilitate the formation of an active transcriptional complex. Evidence indicates that binding of Lrp and Papl to this region inhibits methylation of the GATC site by Dam. However, if this GATC site is first methylated by Dam, binding of Lrp and Papl is inhibited. These events lead to the formation of two different pap methylation states characteristic of active (ON) and inactive (OFF) pap transcription states. The fae (K88), daa (F1845) and sfa (S) pilus operons share conserved ‘GATC‐box’ domains with pap and may be subject to a similar regulatory control mechanism involving Lrp and DNA methylation.
The DNA supercoiling‐sensitive expression of the Salmonella typhimurium his operon requires the his attenuator and is modulated by anaerobiosis and by osmolarity
Authors: O'Byrne Conor P. , Bhriain Niamh Ní , Dorman Charles J.
Published: August 2010

Bacterial cells possess a subset of genes whose expression correlates with changes in DNA supercoiling brought about by anaerobic growth and by growth at high osmolarity. It has been shown previously that expression of the histidine biosynthetic operon of Salmonella typhimurium is derepressed by relaxation of supercoiled DNA. Here, we confirm that a his:: MudJ operon fusion in S. typhimurium can be induced by treatment with the DNA gyrase inhibitor novobiocin in a dose‐dependent manner, and show that the level of derepression is higher in stationary phase than in mid‐exponential phase cultures. Furthermore, expression of his is repressed by anaerobiosis and by osmolarity, two environmental parameters which increase the negative supercoiling of bacterial DNA. Novobiocin induction of his is also repressed by growing the cells either at high osmolarity or anaerobically. Both environmental repression and novobiocin induction of his require the his attenuator. In addition, derepression of his expression by novobiocin and its repression by anaerobiosis or osmolarity are independent of the stringent response gene, relA.
Molecular dissection of structure and function in the lipopolysaccharide of Rhizobium leguminosarum strain 3841 using monoclonal antibodies and genetic analysis
Authors: Kannenberg E. L. , Rathbun E. A. , Brewin N. J.
Published: August 2010

Following treatment with nitrosoguanidine, mutant derivatives of Rhizobium leguminosarum strain 3841 were isolated which failed to react with AFRC MAC 203. This monoclonal antibody normally recognizes a strain‐specific lipopolysaccharide epitope which is developmentally regulated during legume nodule differentiation. Structural modification of lipopolysaccharide (LPS) was analysed by examining reactivity with a range of monoclonal antibodies with different epitope specificities, and also by analysis of LPS mobility changes after electrophoresis on polyacrylamide gels. One class of these LPS‐defective mutants induced normal nitrogen‐fixing (Fix+) nodules on peas (Pisum sativum), while another two classes of Fix‐ mutants were also identified, suggesting that a component of the LPS antigen that is part of the MAC 203 epitope is essential for normal nodule development leading to symbiotic nitrogen fixation. When grown under low‐oxygen or low‐pH culture conditions, one class of Fix‐ mutants completely lacked LPS‐1 (the species that carries O antigen) and a second class showed a modified and truncated form of LPS‐1. Mutants with defective LPS structure were also obtained after Tn5 mutagenesis of R leguminosarum 3841 and all nine Fix‐ mutants were also found to lack the MAC 203 epitope. Three of these transposon‐induced mutants synthesized a truncated form of LPS‐1 that was structurally similar to that of the class of the NTG‐induced mutants described above. These transposon‐induced mutations, and the nitrosoguanidine‐induced Fix‐ mutations, were closely linked and could be suppressed by the same cloned fragment of chromosomal DNA. The data presented here suggest that a precondition for normal nodule development of R. leguminosarum 3841 within pea nodules is the ability to synthesize relatively long‐chain LPS–1 macromolecules under the physiological conditions encountered within the nodule. All mutants that lacked the ability to elongate LPS‐1 macromolecules also failed to express the MAC 203 epitope.
The class 1 outer membrane protein of Neisseria meningitidis produced in Bacillus subtilis can give rise to protective immunity
Authors: Nurminen Marjatta , Butcher Sarah , Idänpään‐Heikkilä IIona , Wahlström Eva , Muttilainen Susanna , Runeberg‐Nyman Kate , Sarvas Matti , Mäkelä P. Helena
Published: August 2010

The class 1 outer membrane protein of Neisseria meningitidis B:15:P1.7, 16 was expressed in Bacillus subtilis in high yield as intracellular aggregates. These were easy to isolate and the protein (called BacP1) could be solubilized under denaturing conditions. Sera of mice immunized with thus‐solubilized BacP1 contained high titres of antibodies that reacted with the class 1 protein of the meningococcal envelope in immunoblots but did not react with native meningococcal envelope in enzyme immunoassays (EIA) or with intact meningococci in bactericidal assays. However, when the BacP1 protein was complexed with heterologous (Salmonella) lipopolysaccharide, the ensuing sera reacted with meningococcal envelope preparations in both EIA and immunoblots, showed subtype‐specific bactericidal activity, and were protective in an infant rat meningitis model.
Sensing of 5′ monophosphate by Escherichia coli RNase G can significantly enhance association with RNA and stimulate the decay of functional mRNA transcripts in vivo
Authors: Jourdan Stefanie S. , McDowall Kenneth J.
Published: August 2010

The RNase E/G family of endoribonucleases has a central role in RNA degradation and processing. Previous work has shown that their cleavage of substrates in vitro can be stimulated by the presence of a 5′ monophosphate group. It has not however, established the importance of this activation for any natural RNA processing or decay pathway in vivo. Here we provide for Escherichia coli RNase G the first evidence that the sensing of a 5′ monophosphate is required in vivo for the normal rapid decay of functional mRNAs; moreover, we show in vitro that, in contrast to a previous study, the presence of a 5′ monophosphate can enhance the affinity of RNase G binding to RNA. The implications of these results along with our finding that the maturation of 16S rRNA is unaffected in cells containing an RNase G mutant impaired in 5′ end sensing are discussed with regard to current models of RNA processing and decay and the molecular mechanism that underlies RNA cleavage by the RNase E/G family.