Burden of gastrointestinal illness in Sweden-SMS as a tool for collecting self-reported gastrointestinal illness.

We collected monthly reports on gastrointestinal illness (GII) episodes among 2348 adults in a 1-year cohort in South West Sweden. The GII episodes were collected by SMS (Short Message System) and validated by telephone interviews among the cohort participants and nationwide. The annual incidence was 0.64 and 0.43 cases per person-year for 28-day self-defined GII (any symptom) and acute GII (vomiting and/or ≥3 episodes of diarrhoea), respectively. The incidence was about 20% higher for the 14-day recall, compared with 28-day recall. The duration of illness was on average 2.3 days. We observed a unimodal seasonal distribution of GII, with the highest prevalence during winter. Responses collected by SMS highly correlated with responses collected by telephone. SMS survey was an efficient tool for the collection of repeated estimates of GII.

Lessons Learnt from the Fukushima Accident-A Swedish Medical Preparedness Perspective.

The triple disaster in March 2011 tragically and severely affected the Japanese society, in spite of its well-developed infrastructure and good access to resources. A multitude of Japanese and international reports have since described the chain of events and actions taken in connection with the earthquake, the tsunami and the power plant failure in Fukushima. In order to further evaluate Japanese experiences of the disaster, and to bring home 'lessons-learnt' of relevance for continued emergency preparedness planning, a group from the National Board of Health and Welfare and other Swedish agencies performed an observer visit to Japan in 2012. A report from the group was recently published. Its main conclusions, and implications focusing on a strengthened national medical preparedness for radionuclear events in Sweden (and possibly elsewhere), are presented here.

Effects of affinity on binding of HER2-targeting Affibody molecules: model experiments in breast cancer spheroids.

Binding of a targeting agent in tumor tissue is influenced by many factors such as molecular weight, charge and affinity of the targeting agent and vascularization of the tumor. In this study, we analyzed tumor cell binding of three HER2-specific and radiolabeled Affibody molecules with different affinities. The Affibody molecules had affinities in the range of 0.12-3.8 nM. Cellular binding was analyzed, after 2 h of incubation, in tumor spheroids composed of BT474 breast cancer cells, which highly express HER2. Binding was, due to the binding-site barrier, limited to the outer 15 ± 5 µm rim of the spheroids, independent of affinity when the concentration of the substances was low. When the concentration was high, the binding site barrier was overcome and the binding occurred approximately 35 ± 5 µm into the spheroids for the two high affinity substances and 50 ± 5 µm for the low affinity substance. The lower affinity might allow for penetration into deeper regions due to less firm binding. We conclude that there is a binding site barrier within tumor spheroids which can be overcome by increased concentration of substance and modified by affinity.

Prevalence and predictive factors of comorbidity in rheumatoid arthritis patients monitored prospectively from disease onset up to 20 years: lack of association between inflammation and cardiovascular disease.

OBJECTIVES: To study the prevalence of comorbid conditions at diagnosis and during follow-up in a cohort of patients with early rheumatoid arthritis (RA) followed prospectively over 20 years, and to identify possible early predictive factors for future comorbidities. METHODS: A community-based cohort of 183 patients (mean age 52 years, 63% female) with early RA was recruited between 1985 and 1989. The presence of comorbidity at RA diagnosis and the occurrence of additional comorbidities were recorded continuously. Possible predictors of future comorbidities were analysed using the Cox proportional hazards model. RESULTS: At RA diagnosis, at least one comorbid condition was present in 43% of the patients. Cardiovascular diseases (CVDs), including hypertension (16% of patients) and malignancy (6% of patients), were most common. In total, 82% of patients developed additional comorbidities during follow-up. CVD and malignancies remained the most frequent comorbidities. Higher age [p < 0.001, odds ratio (OR) 1.03, 95% confidence interval (CI) 1.01­1.15] and the presence of any comorbidity at diagnosis (p = 0.02; OR 1.64, 95% CI 1.08­2.52) predicted future comorbidity. Measures of inflammation at diagnosis or during follow-up were not predictive for development of CVD. CONCLUSION: Comorbidity was present in a considerable proportion of patients in this cohort. More than 40% of patients had another disease at inclusion and during follow-up and > 80% developed additional conditions. The pattern of comorbidity remained unchanged, with CVD and malignancy being most common. Older age and the presence of comorbidity at RA diagnosis predicted the development of comorbidities. The degree of inflammation at any time point was not predictive of future CVD.

Reduced DNAM-1 expression on bone marrow NK cells associated with impaired killing of CD34+ blasts in myelodysplastic syndrome.

Myelodysplastic syndromes (MDS) comprise a heterogeneous group of clonal stem-cell disorders characterized by ineffective hematopoiesis and risk of progression to acute myeloid leukemia. Increased apoptosis and suppressed functions of peripheral blood natural killer (NK) cells have been described in MDS patients, but only limited information is available on the phenotypic and functional integrity of NK cells in the bone marrow. In a cohort of 41 patients with distinct clinical subtypes of MDS, we here show that NK cells in the bone marrow show decreased surface expression of the activating receptors DNAM-1 and NKG2D. Notably, decreased receptor expression correlated with elevated bone marrow blast counts and was associated with impaired NK-cell responsiveness to stimulation with the K562 cell line, or co-activation by NKG2D or DNAM-1 in combination with the 2B4 receptor. Furthermore, antibody-masking experiments revealed a central role for DNAM-1 in NK cell-mediated killing of freshly isolated MDS blasts. Thus, given the emerging evidence for NK cell-mediated immune surveillance of neoplastic cells, we speculate that reduced expression of DNAM-1 on bone marrow NK cells may facilitate disease progression in patients with MDS.

gammaH2AX and cleaved PARP-1 as apoptotic markers in irradiated breast cancer BT474 cellular spheroids.

Chemo- and radiotherapy induce apoptosis in tumours and surrounding tissues. In a search for robust and reliable apoptosis markers, we have evaluated immunostaining patterns of gammaH2AX and cleaved PARP-1 in paraffin-embedded cellular spheroids. Breast cancer BT474 cells were grown as cell spheroids to diameters of 700-800 microm. The spheroids contained an outer cell layer with proliferative cells, a deeper region with quiescent cells and a central area with necrosis. They were irradiated with 5 Gy and the frequency of apoptotic cells was determined at several time points (0-144 h) and distances (0-150 microm) from the spheroids surface. gammaH2AX and cleaved PARP-1 were quantified independently. Apoptotic frequencies for the two markers agreed both temporally and spatially in the proliferative regions of the spheroids. The gammaH2AX signal was stronger and had lower background compared to cleaved PARP-1. The central necrotic region was intensely stained with cleaved PARP-1, whereas no gammaH2AX could be detected. The apoptotic frequency increased with distance from surface for all time points. However, apoptotic frequencies, above unirradiated control levels, could only be detected for the last time point, 144 h after irradiation. We have shown that the spheroid model is a practical system for evaluation of staining patterns and specificities of apoptosis markers. Also, the radial gradient provides the opportunity to study apoptosis under a range of physiological conditions within the same system. We have further shown that gammaH2AX and cleaved PARP-1 are applicable markers for apoptosis in the proliferative regions of the spheroids. However, the more intense and clear staining patterns of gammaH2AX suggests that this marker is preferable for quantification of apoptosis in spheroids and similar paraffin-embedded materials.

Life style habits such as alcohol consumption and physical activity in relation to serum apoB / apoA-I ratio amongst 64-year-old women with varying degrees of glucose tolerance.

OBJECTIVES: To investigate how life style factors such as alcohol consumption and physical activity relate to the serum apoB / apoA-I ratio in a cohort of middle-aged women with varying degrees of glucose tolerance. DESIGN: Observational, cross-sectional cohort study. SETTING: Research laboratory at a University Hospital. SUBJECTS: A screened cohort of 64-year-old postmenopausal women with varying degrees of glucose tolerance, ranging from diabetes (n = 232), impaired (n = 212) and normal (n = 191) glucose tolerance. MAIN OUTCOME MEASURE: ApoB / apoA-I ratio in relation to alcohol consumption and physical activity as assessed by questionnaires. RESULTS: Alcohol consumption and regular physical activity at high levels were inversely associated with the serum apoB / apoA-I ratio independently of confounding factors such as obesity, lipid-lowering treatment, degree of glucose tolerance and hormone replacement therapy. Alcohol seemed related to the apoB / apoA-I ratio mainly through increasing apoA-I, whereas physical activity seemed mainly related to lowering of apoB. Alcohol consumption above a daily intake of 8.9 g, i.e. less than a glass of wine was accompanied by a decrease in apoB / apoA-I ratio. CONCLUSIONS: Amongst these 64-year-old women with varying degrees of glucose tolerance, a moderate alcohol intake and regular physical exercise leading to sweating were associated with lower apoB / apoA-I ratio and these effects seem to be additive.

A randomised placebo controlled 12 week trial of budesonide and prednisolone in rheumatoid arthritis.

OBJECTIVES: To compare budesonide, a locally acting glucocorticoid with minimal systemic exposure, with conventional glucocorticoid treatment and placebo in rheumatoid arthritis. METHODS: A double blind, randomised, controlled trial over 12 weeks in 143 patients with active rheumatoid arthritis, comparing budesonide 3 mg daily, budesonide 9 mg daily, prednisolone 7.5 mg daily, and placebo. Particular attention was paid to the pattern of clinical response and to changes in the four week period following discontinuation of treatment. RESULTS: There were improvements in tender joint count and swollen joint count on budesonide 9 mg compared with placebo (28% for tender and 34% for swollen joint counts, p<0.05). Prednisolone 7.5 mg gave similar results, while budesonide 3 mg was less effective. ACR20 response criteria were met by 25% of patients on placebo, 22% on budesonide 3 mg, 42% on budesonide 9 mg, and 56% on prednisolone 7.5 mg. A rapid and significant reduction in symptoms and signs in response to budesonide 9 mg and prednisolone 7.5 mg was evident by two weeks and maximal at eight weeks. There was no evidence that budesonide provided a different pattern of symptom control from prednisolone, or that symptoms became worse than placebo treatment levels after discontinuation of glucocorticoid treatment. Adverse effects attributable to glucocorticoids were equally common in all groups. CONCLUSIONS: The symptomatic benefits of budesonide 9 mg and prednisolone 7.5 mg are achieved within a short time of initiating treatment, are maintained for three months, and are not associated with any rebound in symptoms after stopping treatment.

Effect of an oral contraceptive on the plasma levels of budesonide and prednisolone and the influence on plasma cortisol.

OBJECTIVE: To investigate whether the use of an oral contraceptive would influence plasma levels of budesonide (Entocort capsules) or prednisolone (plain tablets) during repeated oral administration of these glucocorticosteroids. Plasma concentrations of cortisol and ethinyl estradiol (INN, ethinylestradiol) were also compared. METHODS: Forty healthy women took part in this single-blind, randomized placebo-controlled study with two parallel groups, where a three-way crossover design was applied within groups. One group was taking an oral contraceptive (150 microg desogestrel and 30 microg ethinyl estradiol); the other group (control) was not. On seven consecutive mornings, oral doses of 4.5 mg budesonide, 20 mg prednisolone, or placebo were administered. There was a washout period of at least one menstrual cycle between administration periods. RESULTS: In the oral contraceptive users, the average plasma concentration of prednisolone was 131% higher than in the control group (P < .001), whereas the average plasma concentration of budesonide was only 22% higher (not significant). Mean plasma cortisol levels were suppressed by 90% and 82% with prednisolone and by 22% and 28% with budesonide in oral contraceptive users and the control subjects, respectively. The group difference was significant with prednisolone (P < .001) but not with budesonide. Ethinyl estradiol levels in plasma were not affected by administration of either glucocorticosteroid. CONCLUSION: No difference was found in plasma levels of budesonide or in cortisol suppression after administration of budesonide capsules in women taking the oral contraceptive and those who were not. The oral contraceptive users had much higher plasma levels of prednisolone and greater cortisol suppression. This result suggests that oral budesonide can be used with maintained safety in women using oral contraceptives.

The prevalence of rheumatoid arthritis in Sweden.

The aim of this study was to ascertain the prevalence of rheumatoid arthritis (RA) in a Swedish general adult population. A questionnaire about chronic pain was mailed to a total of 3928 subjects who were chosen as a random sample of the population in two communities in the county of Halland. All persons answering affirmatively to questions intended to identify patients with RA were invited to a clinical examination. X-rays of hands and feet, and analyses of rheumatoid factor and C reactive protein were performed provided that the patients fulfilled two or more of the five clinical items of the 1987 ARA criteria. Furthermore, non-participants were searched for in a patient register and in medical records from the local rheumatology unit in an attempt to identify further cases. Using the modified 1987 ARA criteria for population studies the prevalence rate of RA was calculated to 0.51% (95%, CI = 0.31-0.79).

Recovery from Marie-Bamberger's syndrome and diabetes insipidus after removal of a lung adenocarcinoma with neuroendocrine features.

A 44-year-old woman with Marie-Bamberger's syndrome and diabetes insipidus had a lung tumour with mediastinal metastases, but no signs of metastases to the hypothalamus or pituitary gland. A week after removal of the tumour, the joint pain, polyuria and polydipsia disappeared. The tumour was diagnosed histopathologically as a moderately differentiated adenocarcinoma with focal neuroendocrine cell differentiation and dispersed cells reacting with antisera against neurone-specific enolase, S-100 protein, neuropeptide Y, follicle-stimulating hormone, substance P, vasoactive polypeptide (VIP), adrenocorticotropic hormone and pancreatic polypeptide (PP) as well as to one of three tested antisera raised against antidiuretic hormone (ADH). It was suggested that Marie-Bamberger's syndrome might be caused by one of these immunoreactive substances or by a substance that shares an amino acid sequence with one of these neuroendocrine peptides. It was also suggested that the tumour might produce an ADH-like substance which might have an ADH-antagonist effect.

Segregation properties of bovine papillomaviral plasmid DNA.

Essentially complete segregation of replication-competent BPV-1 plasmid DNA species was observed in daughter subclones derived from primary co-transformed C127 cell lines. Thus, whereas primary co-transformants retained both of two distinguishable co-transfected plasmid species, subcloning experiments revealed that morphologically transformed daughter subclones derived from such co-transformed cell lines contained only one species of viral plasmid DNA. Similar results were obtained with each of two conveniently marked replication and transformation-competent mutants: one with a linker-insertion in the viral upstream regulatory region, and one with a 260 base-pair deletion within the L2 (late) gene, which has no recognized role in plasmid replication or stability. Morphological revertant cell clones that contained no detectable viral plasmid DNA genomes were also isolated at a surprisingly high frequency from clonal wild-type BPV-1 transformed cell lines and from cell lines transformed by various BPV-1 mutants. Further co-transfection experiments were done with a combination of transformation-competent and transformation-defective BPV-1 genomes to investigate a possible role for a viral oncogene in plasmid persistence. In this case, elimination of the transformation-defective mutant was observed after the initial establishment of both input genomes as replicating plasmids in cell clones morphologically transformed by the transformation-competent viral mutant with an intact E5 oncogene. No cell subclones were isolated that contained only the transformation-defective mutant, implying that it was defective in long-term plasmid persistence. Our results indicate that there is significant randomization in the processes of replication and/or partitioning of the BPV-1 genome in mouse C127 cells, and, in combination with previous observations, also suggest that BPV plasmid persistence in C127 cell lines may be the result of a selective proliferative advantage conferred on virus-infected cells by viral oncogene-induced cell growth transformation.

Rolling-circle replication of a high-copy BPV-1 plasmid.

We investigated the replicating form of a bovine papillomavirus type 1 (BPV-1) deletion mutant by direct electron-microscopic analysis of low molecular weight cellular DNA fractions. The detection of viral plasmid DNA replication intermediates was facilitated by the isolation of a spontaneously transformed mouse cell subclone containing an unusually high viral genome copy number (approx. 1000 per cell), and by employing a slight modification of the Hirt fractionation procedure to reduce the level of contaminating linear chromosomal DNA fragments. We observed exclusively rolling-circle-type viral DNA replication intermediates, at a frequency of detection of approximately one replication intermediate per 200 monomeric circular viral DNA molecules. The demonstration of rolling-circles with longer-than-genome-length tails indicated that this high-copy viral plasmid was not subject to a strict once-per-cell-cycle mode of DNA replication. Our observations provide further evidence in favour of an alternative replication mode of the BPV-1 genome, and may help to explain earlier conflicting findings concerning the mechanism of stable BPV-1 plasmid copy-number-control.

Characterization of bovine papillomavirus E1 region deletion mutants associated with neoplastic transformation in a murine cell line.

Spontaneous focus formation in the contact-inhibited C127 cell line, cl.2, harbouring multiple copies of a bovine papillomavirus type 1 deletion mutant, was associated with the evolution of further viral genomic deletions in addition to an amplification of the viral genome copy number. Three simple frameshift deletions of 308, 605 and 1291 bp, associated with separate transformation events, were mapped within the E1 open reading frame, implying a common mechanism of spontaneous transformation in this cell line. Furthermore, each transformed cell line also retained multiple copies of the intact E1 gene, suggesting that these novel deletion mutants might function by a dominant-negative mechanism to disrupt the normal control of viral DNA replication or viral transcription. These mutants had the potential to encode truncated E1 polypeptides with a common N-terminal region encoded by the 5' end of E1, i.e. overlapping the previously described E1 modulator gene. A possible role for these mutants in diverting a lethal type of virus-cell interaction is discussed.

Carcinoid granule extract contains antigenic determinants common to peptide hormone-producing cells and endocrine tumours.

Some soluble proteins, such as the chromogranins, are found in nearly all peptide hormone-producing cells. Little is known about their functional role, although they may act as enzymes or represent structural proteins. In the present study we have isolated granules from an ileal carcinoid tumour and raised antibodies to protein constituents within them. The antiserum proved to be useful for the immunohistochemical demonstration of peptide hormone-producing endocrine tumours in general and for the demonstration of most peptide hormone-producing cells.

Histology of gastric biopsies from peptic ulcer patients before and after short-term treatment with omeprazole or H2-receptor antagonists.

Sixty patients with duodenal or prepyloric ulcers were given omeprazole (30 or 40 mg o.m.; average period of treatment: 2.9 weeks) or histamine H2-receptor antagonists (cimetidine 400 mg b.i.d. or ranitidine 150 mg b.i.d.; average period of treatment; 3.5 weeks) for a single period ranging between 2 and 6 weeks. At the end of the treatment period fasting plasma gastrin levels were moderately increased in both groups in comparison with the pretreatment values. Endoscopic biopsies were taken from the oxyntic mucosa at the beginning and at the end of the treatment period. Light microscopy of the biopsies was aimed particularly at determining the number of endocrine cells. In addition, the mucosal thickness and the volume densities of the parietal cells, the lamina propria and the gland lumina were measured. There were no significant differences in the endocrine or parietal cell populations, between biopsies taken from the patients before and after treatment with omeprazole or histamine H2-receptor antagonists. The mucosal thickness and the densities of the lamina propria and of the gland lumina remained unaffected by the treatment.

Endocrine cells in the human oxyntic mucosa. A histochemical study.

The oxyntic mucosa of the human stomach harbors at least five different endocrine cell types (ECL cells, A-like or X cells, somatostatin cells (D), enterochromaffin (EC) cells, and D1 or P cells). Little is known about their functional roles, and of the hormones they produce only somatostatin has been identified. The relative frequency and regional distribution of the different endocrine cell populations were studied in 13 adults with no manifest gastrointestinal disease. From each of them at least three biopsy specimens were taken at seven fixed locations within the oxyntic mucosa. The specimens were examined for the different endocrine cell types by means of immunocytochemistry (staining with antisera against chromogranin A,5-hydroxytryptamine, and somatostatin) and silver staining techniques (demonstration of argyrophil cells by the methods of Grimelius or Sevier-Munger). Chromogranin-positive cells included all endocrine cells identified by the other staining techniques. Grimelius-positive cells included all endocrine cells except the somatostatin cells. Sevier-Munger-positive cells, finally, included the ECL cells and the EC cells. The frequency of ECL cells could be calculated by subtracting the number of EC cells from the number of Sevier-Munger-positive cells. The ECL cells represented 35% of the total endocrine number, somatostatin cells 26%, and EC cells 25%. The remaining 14% consisted of A-like cells, D1 cells, and P cells. Generally, the endocrine cells predominated in the basal portion of the glands, but the various populations of endocrine cells were not uniformly distributed in the various regions of the oxyntic mucosa. However, representative specimens could be obtained from the main body of the stomach, and the results indicate that the examination of a fairly small number of specimens from the main body of the stomach may be sufficient for assessing the frequency of endocrine cells in the oxyntic mucosa of individual patients.