Redefining high-risk patients with stage II colon cancer by risk index and microRNA-21: results from a population-based cohort.

BACKGROUND: The aim of the present study was to analyse the prognostic value of microRNA-21 (miRNA-21) in patients with stage II colon cancer aiming at a risk index for this group of patients. METHODS: A population-based cohort of 554 patients was included. MicroRNA-21 was analysed by qPCR based on tumour tissue. An index was created using the coefficients obtained from a collective multiple Cox regression. The entire procedure was cross-validated (10-fold). The performance of the index was quantified by time-dependent receiver operating characteristics curves. RESULTS: High miRNA-21 expression was associated with an unfavourable recurrence-free cancer-specific survival (RF-CSS), hazard ratio 1.35 (95% confidence interval, 1.03-1.76) (P=0.028). The generated RF-CSS index divided the traditional high-risk patients into subgroups with 5-year RF-CSS rates of 87% and 73%, respectively (P<0.001). The overall survival (OS) index identified three different subgroups (P<0.001). Cross-validated 5-year OS rates were 88%, 68%, and 50%, respectively. CONCLUSIONS: This population-based study supports miRNA-21 as an additional prognostic biomarker in patients with stage II colon cancer. Furthermore, the introduction of a risk index may guide the use of postoperative adjuvant treatment in a more appropriate way compared with current practice.

Obesity and nocturnal gastro-oesophageal reflux are related to onset of asthma and respiratory symptoms.

Several studies have identified obesity as a risk factor for asthma in both children and adults. An increased prevalence of asthma in subjects with gastro-oesophageal reflux (GOR) and obstructive sleep apnoea syndrome has also been reported. The aim of this investigation was to study obesity, nocturnal GOR and snoring as independent risk factors for onset of asthma and respiratory symptoms in a Nordic population. In a 5-10 yr follow-up study of the European Community Respiratory Health Survey in Iceland, Norway, Denmark, Sweden and Estonia, a postal questionnaire was sent to previous respondents. A total of 16,191 participants responded to the questionnaire. Reported onset of asthma, wheeze and night-time symptoms as well as nocturnal GOR and habitual snoring increased in prevalence along with the increase in body mass index (BMI). After adjusting for nocturnal GOR, habitual snoring and other confounders, obesity (BMI >30) remained significantly related to the onset of asthma, wheeze and night-time symptoms. Nocturnal GOR was independently related to the onset of asthma and in addition, both nocturnal GOR and habitual snoring were independently related to onset of wheeze and night-time symptoms. This study adds evidence to an independent relationship between obesity, nocturnal gastro-oesophageal reflux and habitual snoring and the onset of asthma and respiratory symptoms in adults.

Cloning, sequence analysis and functional characterization of DNA polymerase I from the thermophilic eubacterium Rhodothermus marinus.

A gene encoding a DNA polymerase I from the thermophilic eubacterium Rhodothermus marinus was identified. The gene was cloned, sequenced and expressed in Escherichia coli. The gene is 2772 bp long and encodes a protein of 924 amino acids with a calculated molecular mass of 104.8 kDa. Sequence analysis showed that a generally conserved Phe residue in the O-helix is substituted by a Tyr (position 756) in the R. marinus enzyme. A Tyr in this position decreases the discrimination against dideoxynucleotides which is a major advantage in DNA sequencing. The protein was purified, characterized and showed to contain specific DNA-polymerization activity of 3100 units/mg of protein, 5'-->3' exonuclease activity and a 3'-->5' proofreading activity. Its optimum activity was at 55 degrees C and it had a half-life of 2 min at 90 degrees C. A truncated form of the enzyme lacking the 5'-->3' exonuclease domain was also expressed in E. coli. It had a specific DNA-polymerization activity of 5000 units/mg of protein and lacked the 5'-->3' exonuclease activity. Its optimum activity was at 65 degrees C and it had a half-life of 11 min at 90 degrees C. It was usable for DNA sequencing. This is the first thermostable DNA polymerase described with the O-helix Phe-->Tyr substitution.

Long PCR detection of the C4A null allele in B8-C4AQ0-C4B1-DR3.

The genes coding for the two components of complement 4 (C4), C4A and C4B, are located within the major histocompatibility complex (MHC) on the short arm of chromosome 6. Several studies have shown that deficiency of C4A is associated with systemic lupus erythematosus (SLE), rheumatoid arthritis and scleroderma. A large deletion covering most of the C4A gene and the 21-hydroxylase-A (21-OHA) pseudogene found on the extended haplotype B8-C4AQ0-C4B1-DR3 is estimated to account for approximately two-thirds of C4A deficiency in Caucasian SLE patients. Detection of this C4A null allele has been technically difficult due to the high degree of homology between C4A and C4B, with protein analysis and restriction fragment length polymorphism (RFLP) analysis using Southern blotting being the only approaches available. In this study, a long PCR strategy was used to rapidly genotype for the C4A deletion through specific primer design. The methodology makes use of the unique sequence of the G11 gene upstream of C4A and the sequence of a 6.4 kb retrotransposon, the human endogenous retrovirus HERV-K(C4), which is present in intron 9 of C4A but absent in the case of the deletion.

Cloning, sequence analysis and overexpression of a rhodothermus marinus gene encoding a thermostable thymidine kinase.

Thymidine kinase type II is an important part of the pyrimidine salvage pathway. The thymidine kinase gene from the thermophilic eubacterium Rhodothermus marinus was cloned, sequenced and overexpressed. The gene is 639 bp and encodes a protein of 213 amino acids with a calculated molecular mass of 23.6 kDa. It shows homology to other thymidine kinase proteins from eukaryotic and prokaryotic organisms. The recombinant protein is inhibited by dNTPs but not by dNDPs. It is a tetramer in its native state. Its optimum temperature of activity is 65 degrees C and it has a half life of 15 min at 90 degrees C. This is the first thymidine kinase to be described from a thermophilic bacterium.

Effects of nicotine nasal spray on atherogenic and thrombogenic factors during smoking cessation.

OBJECTIVES: To investigate changes in cardiovascular risk factor parameters when stopping smoking and to identify any impact of nicotine nasal spray on these factors. DESIGN AND SUBJECTS: In a placebo-controlled, double-blind 3-month prospective study, nicotine nasal spray (NNS) or a placebo was given to 157 subjects attending a smoking cessation programme. Blood samples from 46 subjects who remained abstinent for 3 months were analysed. Nasal spray use was given on an ad libitum basis. RESULTS: The haemoglobin (Hb) decreased from 149.0 to 143.2 g L(-1) (P<0.001). The haematocrit (Hct) decreased from 44.6 to 42.4% (P<0.001). The mean corpuscular volume (MCV) decreased from 93.4 to 92.3 fl (P<0.001). The mean corpuscular haemoglobin concentration (MCHC) increased from 333.9 to 338.1 g L(-1) (P = 0.029). The white blood cell count (WBC) decreased from 8.4 to 6.6x10(9) L(-1) (P<0.001). The total cholesterol decreased from 5.92 to 5.65 mmol L(-1) (P = 0.015). The high-density lipoprotein cholesterol (HDL) increased from 1.29 to 1.44 mmol L(-1) (P = 0.48) and low-density lipoprotein cholesterol (LDL) decreased from 4.00 to 3.54 mmol L(-1) (P = 0.004). The HDL/LDL ratio increased from 0.36 to 0.46 (P = 0.011). CONCLUSION: Stopping smoking resulted in positive effects on cardiovascular risk factors. Nicotine treatment for as long as 3 months did not have any significant effect on these 'stopping smoking changes'. In smoking cessation, nicotine substitution up to 3 months seems to be safe.

Nicotine nasal spray with nicotine patch for smoking cessation: randomised trial with six year follow up.

OBJECTIVE: To evaluate the efficacy of using a nicotine patch for 5 months with a nicotine nasal spray for 1 year. DESIGN: Placebo controlled, double blind trial. SETTING: Reykjavik health centre. SUBJECTS: 237 smokers aged 22-66 years living in or around Reykjavik. INTERVENTIONS: Nicotine patch for 5 months with nicotine nasal spray for 1 year (n=118) or nicotine patch with placebo spray (n=119). Treatment with patches included 15 mg of nicotine for 3 months, 10 mg for the fourth month, and 5 mg for the fifth month, whereas nicotine in the nasal spray was available for up to 1 year. Both groups received supportive treatment. MAIN OUTCOME MEASURE: Sustained abstinence from smoking. RESULTS: Sustained abstinence rates for the patch and nasal spray group and patch only group were 51% v 35% after 6 weeks (odds ratio 1.97, 95% confidence interval 1.17% to 3.32; P=0.011(chi2), 37% v 25% after 3 months (1.76, 1.01 to 3.08; P=0.045), 31% v 16% after 6 months (2.40, 1.27 to 4.50; P=0.005), 27% v 11% after 12 months (3.03, 1.50 to 6.14; P=0.001), and 16% v 9% after 6 years (2.09, 0.93 to 4.72; P=0.08) [corrected]. CONCLUSIONS: Short and long term abstinence rates show that the combination of using a nicotine patch for 5 months with a nicotine nasal spray for 1 year is a more effective method of stopping smoking than using a patch only. The low percentage of participants using the nasal spray at 1 year, and the few relapses during the second year, suggest that it is not cost effective to use a nasal spray for longer than 7 months after stopping a patch.

The effects of fluoxetine combined with nicotine inhalers in smoking cessation--a randomized trial.

AIMS: Nicotine replacement therapy (NRT) is an established aid in stopping smoking, while the role of antidepressants remains uncertain. Antidepressants added to NRT might improve abstinence rates. Our aim was to determine the efficacy of nicotine inhaler and fluoxetine vs. nicotine inhaler and placebo in attempts to quit smoking. DESIGN: A randomized, double-blind, placebo-controlled trial. SETTING: A smoker's cessation clinic. PARTICIPANTS: One hundred volunteers smoking 10 cigarettes/day or more. INTERVENTIONS: Subjects were instructed to start taking a daily dose of 10 mg of fluoxetine or placebo 16 days before stopping smoking, then 20 mg 10 days before quitting, continuing for up to at least 3 months. Subjects were instructed to use 6-12 units per day of nicotine inhalers after stopping smoking for up to 6 months. MEASUREMENTS: Continuous abstinence rates recorded at various time points up to 12 months from the quit date. FINDINGS: The sustained abstinence rate for the inhaler-fluoxetine group was 54%, 40%, 29% and 21% after 1.5, 3, 6 and 12 months, respectively, compared to 48%, 40%, 32% and 23% for the inhaler-placebo group. The differences were not significant at any time point. Abstinence up to 3 months was more likely in older smokers, those with a lower Beck Depression Inventory Score (BDI), lower Fagerström Test of Nicotine Dependence (FTND) score and no history of alcoholism. Fluoxetine appeared to increase abstinence rates among high BDI smokers compared to high BDI smokers assigned placebo. Serum levels of nicotine during treatment in the inhaler-fluoxetine group were lower than in the inhaler-placebo group so that fluoxetine may have reduced inhaler use through a common site of action. CONCLUSIONS: We found no evidence that fluoxetine treatment when used as an adjunct to NRT in unselected smokers is effective, but there may be an advantage to using it in depressed smokers.

A double-blind randomized trial of nicotine nasal spray as an aid in smoking cessation.

The objective of the study was to evaluate the therapeutic efficacy of nicotine nasal solution (NNS) for smoking cessation from the stopping day up to 3 months. We also followed the participants for 2 yrs after ceasing smoking to assess what happens after stopping using NNS. In a placebo-controlled, double-blind, 2 yr prospective study, 157 smokers were given either NNS, one dose containing 1 mg of nicotine per 100 microL (n=79), or placebo (n=78). Treatment was continued for up to 1 yr. One day after quitting smoking, the average number of daily doses was 11 in the group assigned NNS and 14 in the group assigned the placebo, and after 6 weeks, 14 and 6 doses, respectively, among abstinent participants still using spray. After 3 months, 65% of the abstainers in the nicotine group were still using the NNS. The abstinence rates were 51, 39 and 29% after 6 weeks, 3 and 6 months, respectively, as compared to 24, 19 and 18% in the placebo group (p=0.0003; p=0.003; p=0.050). The proportion abstinent at the 1 yr (25 vs 17%) and 2 yr follow-ups (19 vs 14%) was higher among those assigned to the nicotine than to the placebo group, but not significantly so for the numbers used in the study. In conclusion, the use of nicotine nasal spray significantly increased the abstinence rate during the first 6 months following the quitting day.

[Tuberculous infection and tuberculosis in the foreign born in Iceland.].

OBJECTIVE: The purpose of the study was to find the incidence and prevalence of tuberculosis among the foreign-born in Iceland. MATERIAL AND METHODS: The study material was obtained from (a) the National TB register and (b) the files of the Immigration Office on recidency applicants in 1995. RESULTS: In 1975-1996 there were 32 cases of TB in the foreign-born out of a total of 468. The proportion of cases among the foreign-born rose significantly during the period (p<0.001). In 22 years the incidence of TB among the foreign-born was 18.0 but 8.4 among those born in Iceland (p<0.001). The incidence of TB in Asian-born was 173.7, or 21 times that among those born in Iceland (p<0.001). The second highest incidence (18.8) was in those born in North and South America. Tuberculosis usually appeared within five years of immigration. During 1995 the 559 applicants for recidence permit provided health certificates. Of these 363 had a tuberculin skin test (TST) and 42% were positive. The corresponding figure for those born in Africa was 68%, in Asia 58% and in East Europa 50%. Most of those with positive TST had a chest x-ray but also 23% of the others. 26.2% had neither a TST nor a chest x-ray but still received a health certificate. Only 33% of those positive received isoniazide to eliminate infection and 88% of these completed at least six months of isoniazide treatment. CONCLUSION: A TST is an indispensable part of health screening for immigrants and also a chest x-ray when appropriate. Treatment of TB infection should be used more often.

[Bronchial asthma and respiratory symptoms among Icelanders 20-44 years of age.].

OBJECTIVE: To estimate the prevalence of asthma and respiratory symptoms in an urban population. MATERIAL: Eight hundred men and women aged 20-44 years, living in the capital Reykjavik and suburbs. METHODS: Participants answered a questionnaire, underwent skin prick testing for atopy, spirometry and a test for bronchial hyperresponsiveness (BHR) by methacholine challenge. RESULTS: There was 77% attendance. Altogether 16.6% reported wheezing or whistling at any time in the last 12 months. Altogether 32 (5.6%)answered yes to "Have you ever had asthma?" and the diagnosis had been confirmed by a doctor in all but four. Fourteen (2.5%) had suffered from an attack of asthma in the last 12 months wheras only 0.9% were currently using anti asthmatic drugs. BHR was found among 8.7% and atopy on skin testing among 20.5%. BHR was more common among those with airflow obstruction and three times more common among the atopic participants (18% vs. 6%, p%lt;0.01). By using a history of wheezing during the last 12 months together with BHR and/or a history of doctor confirmed asthma the prevalence of current asthma was found to be 5% in our sample. The main predictive factors for asthma were a history of breathlessness and nighttime breathing symptoms, but also atopy, airflow obstruction and a maternal history of asthma. CONCLUSION: Even by using a conservative defination, asthma is a common disorder among 20-44 years old Icelanders whereas the use of asthma medication is rather uncommon in this population.

[Incidence and prevalence of positive tuberculin skin test reactions among schoolchildren.].

OBJECTIVES: To investigate the incidence and prevalence of positive tuberculin skin test reactions in schoolchildren from six to 16 years of age. MATERIAL AND METHODS: Data from tuberculin test school surveys in Reykjavik during the years 1958 to 1991 are available for almost all individuals of each age cohort. During 1958 to 1991 schoolchildren from six to 16 years of age were tested annually. In Iceland regular BCG vaccination in children or aldults has never been applied. RESULTS: Incidence of positive tests in all ages fell from 2.5 (per 1000 tested per year) from 1958 down to 0.5 in the mid seventies and after that the incidence remained low. Prevalence of positive tests for the same age group in different age cohorts showed that for each age group from seven to 16 years positive tests were most common at the beginning of the study period but decreased successively to the beginning of the seventies. The prevalence of positive tests was low and almost unchanged during 1976 to 1991 and varied from 0 -1.6 (per 1000 tested per year) among children seven to eight years of age, 0-2.9 among children 11 to 12 years of age and from 0-3.8 among those aged 15 to 16 years. CONCLUSION: During the last decade little has been gained by systematic testing for tuberculosis as a tool for finding newly infected persons or carriers of M. Tuberculosis. It seems to us that the results of this study do not justify systematic testing for tuberculous infection in all schoolchildren. Increased emphasis should be placed on testing among risk groups such as immigrants from countries where tuberculosis is endemic. Recent infection due to M. Tuberculosis is a high risk factor for tuberculosis. Search for newly infected persons in close contact with infectious patients with tuberculosis should have priority next to the diagnosis and treatment of the patients. Tests for tuberculosis among those who want to immigrate to Iceland circumscribe another risk group where containment of tuberculosis is possible.

[Tuberculosis control in Scandinavia]. / Tuberkulosbekämpning i Norden.

Compared with the rest of the world, the situation regarding tuberculosis is favourable in the Nordic countries. In 1993, the incidence for persons born in respective country (per 100,000 of the population) was 4.1 in Denmark, 3.2 in Iceland and Sweden, 3.8 in Norway and 10.0 in Finland. The respective average figures for foreign-born residents were 6-14 times higher. Programmes for contending with tuberculosis have been uniform in the Nordic countries, with the exception of BCG (bacillus Calmette-Guérin) vaccination policy, though childhood tuberculosis is rare in the area. However, tuberculosis represents a manifest problem among the foreign-born population, first and foremost among the increasing number of refugees.

[Atopic allergy in an Icelandic urban population 20-44 years of age.].

This is a part of a second stage of EC Respiratory Health Survey and the aim of this study is to estimate the prevalence of atopic allergy among urban population of men and women aged 2CM4 years, living in the capital Reykjavik and suburbs. Altogether 800 individuals were invited of whom 570 participated (77%). All participants answered questionnaire and 540 individuals were skin prick tested by 12 allergens. 20.5% had positive skin tests, defined as at least one allergen positive by a weal reaction of 5=3 mm, 20.1% male and 20.9% female. The highest prevalence of allergy was in the agegroup 20-24 years. Results of single allergens: timothy grass 8.5%, cat 7.6%, dog 6.3%, D. pteronyssinus 6.1%, Lepidoglyphus destructor 3.2%, birch 3.0% and Cladosporium 1.1%. Other allergens reacted positive in less than 1% each; altogether 11.2% were positive to one or more animals and 10.6% to timothy and/or birch. RAST tests were done by five allergens. RAST results =50.35 ku/1 (RAST class 5=1) to timothy was 11.9%, to cat 7.5%, to D pteronyssinus 9.2%, to birch 5.9% and to cladosporium 6.5%. The only statistically significant risk factor for allergy was mothers history of allergy symptoms. Compared to already published data from other EC Respiratory Health Survey countries, the prevalence of atopic allergy appear to below in Iceland.

Controlled trial of nicotine polacrilex gum with supportive measures.

In a placebo-controlled, double-blind, 2-year prospective study, 182 smokers were given either nicotine polacrilex gum containing 4 mg of nicotine (n = 92) or a placebo (n = 90). The number of participants abstinent at 2-year follow-up was 41 (44.6%) of 92 in the nicotine group vs 28 (31.1%) of 90 in the placebo group. Abstinence rates for daily nicotine gum users (n = 64) at 12 months and again at 24 months remained 48.4%, as compared with 26.1% and 31.9% for the daily placebo gum users (n = 69). Of participants with a high nicotine-dependence score, those allotted to the nicotine group rather than to the placebo group were 13 times more likely to be abstaining at the 2-year follow-up. Use of nicotine polacrilex gum, therefore, can substantially aid in stopping smoking, particularly among highly dependent smokers.

Nuclear DNA in carcinoid tumours of the lung.

Fluorescence cytophotometric DNA measurements on 15 carcinoid tumours generally yielded histograms with a prominent G1-phase peak and only low frequencies of S- and G2-phase nuclei. In 13 tumours the modal DNA value was near diploid (1.75-2.25). Three of these 15 tumours showed a much greater number of S- and G2-phase values than the others and in two of them there were also G1-phase peaks below the near-diploid range. Among 22 samples of small cell carcinoma of the lung (SCCL), the modal DNA value was hypodiploid in two, near diploid in seven and hyperdiploid in 13. The proportion of S-phase cells was higher in the SCCL samples than in the carcinoid tumours. It is suggested that the nuclear DNA distribution profile may facilitate morphological discrimination of a classical pulmonary carcinoid from small cell carcinoma. In a borderline case, the finding of a hyperdiploid DNA stem line and high proliferative activity will favour the diagnosis of SCCL.

An unnecessary case of paraffin oil pneumonia.

A 64-year-old woman died after 12 years of progressive pulmonary disease which was initially diagnosed as sarcoidosis but later correctly identified as mineral oil pneumonia due to insufflation of paraffin instilled as nasal drops. In view of the potential damaging effect of liquid paraffin on the lungs, the current indications for its use must be strongly questioned.

DNA ploidy in small cell carcinoma of the lung.

Fluorometric single cell DNA analysis was performed on 123 tumour samples from 33 patients who had died of small cell carcinoma of the lung (SCCL). In 36% (12 patients) the modal DNA profile was hypodiploid or near diploid and the median survival time was 16.3 months, as opposed to 9.5 months for the 21 patients with hyperdiploid DNA profile, who also seemed to have a lower response rate to chemotherapy. It is suggested that an estimation of the modal DNA ploidy before the start of the treatment in SCCL might be used as a cellular parameter to obtain more comparable treatment study groups and for the prediction of survival.