Assessing amyotrophic lateral sclerosis prevalence in Norway from 2009 to 2015 from compulsory nationwide health registers

<p><i>Objective</i>: In Norway, diagnoses from specialist health care visits, drug prescriptions, and causes of deaths are registered in compulsory health registers. We aimed to determine amyotrophic lateral sclerosis (ALS) prevalence from 2009 to 2015 by combining these registers. <i>Methods</i>: We validated the Norwegian Patient Registry (NPR) through hospital files, and linked it with the Norwegian Cause of Death Registry and the Norwegian Prescription Database. Poisson regression models were fitted for estimating gender ratios, time trends and possible interactions. Similar models were used for mortality data subtracted from the dataset. <i>Results</i>: Eleven percent of patients with at least one ALS-related entry in NPR did not have ALS. ALS prevalence could nevertheless be reliably estimated through ascertaining cases identified in two separate registers, or with at least two entries in NPR with first entry within four years prior to prevalence date. ALS prevalence remained stable, and was 7.6/100,000 (95% CI 6.9–8.4) at 31st December 2015. Mean male:female ratio was higher for prevalence (1.8; 95% CI 1.6–2.0) than for mortality (1.5; 95% CI 1.2–1.8) (<i>p</i> = 0.04). There were also significant regional differences in prevalence (<i>p</i> < 0.01) but not in mortality. <i>Conclusions</i>: Norwegian compulsory health registers provide reliable tools for ALS surveillance, and suggest gender and regional differences in survival after diagnosis.</p

A One Year Follow-Up Study of Natural Killer and Dendritic Cells Activities in Multiple Sclerosis Patients Receiving Glatiramer Acetate (GA)

<div><p>Background</p><p>Multiple sclerosis (MS) is a chronic inflammatory, demyelinating and neurodegenerative disease. It is thought to be mediated by CD4⁺ Th1/Th17 cells. More recently, cells of the innate immune system such as dendritic cells (DCs) and natural killer (NK) cells have been in focus. Glatiramer acetate (GA) is an approved drug for treating MS patients.</p><p>Methodology/Principal Findings</p><p>In the current study we examined the activities of NK and DCs in nine relapsing remitting MS patients for up to one year after initiation of GA treatment. We observed that NK cells isolated from most of these patients have increased cytotoxic activity against K562 cells. Further analysis showed that the same NK cells lysed both autologous immature (i) and mature (m) DCs. In most patients this increased activity was correlated with increased NK cell activating cytotoxicity receptors such as NKp30, NKp44, NKp46 and NKG2D, and reduced expression of the inhibitory molecule CD158 on the surface of these NK cells. The expression of HLA-DR was increased on iDCs and mDCs in the majority of the patients, but no consistency was observed for the expression of HLA-I or HLA-E. Also, the co-stimulatory receptors CD80, CD83 or CD86 expression was down-regulated on iDCs and mDCs in most cases. Further, the expression of CCR6 was increased on mDCs at later time points of therapy (between 32–48 weeks).</p><p>Conclusions/Significance</p><p>Our results are the first showing the effects of GA treatment on NK cells in MS patients, which may impact future use of this and other drugs to treat this disease.</p></div

Clinical and demographic data of the relapsing remitting multiple sclerosis patients included in the study.

1<p>Two or more oligoclonal bands in CSF at isoelectric focusing that were not present in serum.</p>2<p>Number of weeks after treatment starts when blood samples were taken.</p>3<p>Patient 2 had previously used interferon-β1a treatment, the others were treatment naive.</p><p>Abbreviations: GA = glatiramer acetate, M = Male, F = Female, ON = Optic Neuritis, EDSS = Expanded disability status scale, OCB = Oligoclonal Bands, ND = No data.</p

GA treatment of RRMS patients increases NK cell lysis of tumor target cells.

<p>NK cells were isolated from MS patients either pre-treatment (Pre) or after several weeks of treatment (ranging from 1–48 weeks). These cells were activated in vitro for five days and then examined for their ability to lyse the leukemic cell line K562. Percent cytotoxicity is shown as mean ± SEM of four parallel experiments done from cells isolated from the same patient. P values compared lysis of tumor cells by NK cells isolated from GA-treated patients before (Pre) and weeks after treatment.</p

Treatment with GA changes expression of surface molecules in iDCs from RRMS patients.

<p>The expression of HLA-DR, HLA-I, HLA-E as well as the co-stimulatory molecules CD80, CD83 or CD86 was analyzed by flow cytometry of iDCs from RRMS patients before (pre) and weeks after treatment-start. Mean fluorescence intensity (MFI) of 10000 analyzed cells is shown.</p

IL-2 activated NK cells from MS patients treated with GA show increased expression of activating cytotoxicity receptors.

<p>NK cells from MS patients were activated with IL-2 in vitro for five days and then examined for the expression of activating receptors NKp30, NKp44, NKp46 and NKG2D, as well as the inhibitory receptor CD158. Figures show comparisons of receptor expression before (Pre) and weeks after treatment start with GA. Mean fluorescence intensity (MFI) of 10,000 analyzed cells is shown.</p

Assessments of experimental variability of the cytotoxicity assays.

<p>NK cells were isolated from three healthy individuals. These cells were activated in vitro for five days and then examined for their ability to lyse iDCs and K562 tumor cells. Percent cytotoxicity is shown as mean ± SEM. A) and B) show assays done from two healthy donors with two different E:T ratios. C) and D) show the results from a third healthy donor with repeated samples at baseline, or 3 and 8 weeks later.</p

GA treatment of RRMS patients increases NK cell lysis of autologous monocyte-derived iDC and mDC target cells.

<p>NK cells were isolated from MS patients either pre-treatment (Pre) or weeks after treatment (ranging from 1–48 weeks). These cells were activated in vitro for five days and then examined for their ability to lyse iDCs. Percent cytotoxicity is shown as mean ± SEM of four parallel experiments done on cells isolated from the same patient. P values compared lysis of tumor cells by NK cells isolated from GA-treated patients before (Pre) and weeks after treatment.</p

GA treatment of RRMS patients increases NK cell lysis of autologous monocyte-derived mDC target cells.

<p>NK cells were isolated from MS patients either pre-treatment (Pre) or weeks after treatment (ranging from 1–48 weeks). These cells were activated in vitro for seven days and then examined for their ability to lyse mDCs. Percent cytotoxicity is shown as mean ± SEM of four parallel experiments done on cells isolated from the same patient. P values compared lysis of tumor cells by NK cells isolated from GA-treated patients before (Pre) and weeks after treatment.</p

Differences in urine NTX between patients and controls, without and with adjustment for possible confounders.

<p>β: unstandardized β coefficient, CI: β confidence interval. NTX (cross-linked N-terminal telopeptide of type 1 collagen) was analyzed as dependent and the following as independent variables (separately and simultaneously): recent childbirth, breastfeeding, systemic glucocorticoids, current smoking, alcohol use, began menopausal transition, sun tanning in northern latitudes, and phosphate. Recent childbirth and breastfeeding were not included simultaneously in the regression analysis due to their high correlation (r = 0.921; p<0.001).</p