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Bettina Ewers,1 Mette Rosenlund Sørensen,2 Sisse Fagt,2 Lars J Diaz,1 Tina Vilsbøll1,3 1Clinical Research, Steno Diabetes Center Copenhagen, Gentofte, Denmark; 2The National Food Institute, Technical University of Denmark, Søborg, Denmark; 3Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, DenmarkCorrespondence: Bettina EwersClinical Research, Steno Diabetes Center Copenhagen, Niels Steensens Vej 2, Gentofte, DK-2820, DenmarkTel +45 3091 2997Email bettina.ewers@regionh.dkPurpose: Intention and perceptions of healthy eating may affect diet-related behavior. We assessed the intention and perceptions of eating healthily in patients with type 1 (T1D) and type 2 diabetes (T2D) compared with the general population. Secondly, differences in diet quality were assessed in patients with diabetes perceiving their dietary habits as more or less healthy.Materials and Methods: This cross-sectional study included data on socioeconomic status, dietary intake, and questions on healthy eating from adults with T1D (n=426), T2D (n=348) and from the general population (n=2899).Results: Patients with T2D were less likely to perceive their dietary habits as healthy compared with T1D and the general population. Patients with T1D or T2D perceiving their dietary habits as healthy reported higher intake of vegetables, fruit, fish, fibre and protein. In addition, patients with T1D with perceived healthy versus less healthy dietary habits had lower sugar intake and higher alcohol intake. Overall, adherence to dietary guidelines in patients with T1D and T2D was too low both in self-perceived healthy and less healthy eaters. In comparison with T1D patients, patients with T2D were less likely and the general population was more likely to strive to eat a healthy diet.Conclusion: Patients with T2D had poorer self-perception of their dietary healthiness and less intention of eating healthily, compared with patients with T1D and the general population. Actual diet quality was higher amongst patients with T1D and T2D perceiving their dietary habits as healthy than those perceiving their dietary habits as less healthy. But inadequate intakes were found in all groups. Health care providers should address and explore the patient’s intention and perceptions of healthy eating when discussing dietary changes in diabetes to improve nutritional support.Keywords: diabetes, dietary adherence, dietary intake, nutrition

This case report describes a 29-year-old man, who was admitted to a psychiatric hospital due to a severe depressive episode without psychotic symptoms. After two weeks he developed acute retrograde autobiographical amnesia. No organic cause was identified, and the patient was therefore diagnosed with dissociative amnesia. The depressive symptoms ceased as the amnesia developed. After five months of follow-up in the outpatient clinic, his amnesia for the time preceding its outbreak remained unchanged. He patient managed to resume a functional daily life., (Published under Open Access CC-BY-NC-BD 4.0. https://creativecommons.org/licenses/by-nc-nd/4.0/.)

Aim: To evaluate whether both bolus insulin injection frequency and smart pen engagement were associated with changes in glycaemic control, using real-world data from adults with type 1 diabetes (T1D)., Materials and Methods: Adults using a smart pen (NovoPen 6) to administer bolus insulin (fast-acting insulin aspart or insulin aspart) alongside continuous glucose monitoring were eligible for inclusion. Smart pen engagement was characterized by number of days with pen data uploads over the previous 14 days. Glycaemic control was evaluated by analysing glucose metrics., Results: Overall, data from 1194 individuals were analysed. The number of daily bolus injections was significantly associated with time in range (TIR; 3.9-10.0 mmol/L [70-180 mg/dL]; P < 0.0001). Individuals administering, on average, three daily bolus insulin injections had an estimated 11% chance of achieving >70% TIR. The probability of achieving >70% TIR increased with the mean number of daily bolus injections. However, the percentage of TIR was lower on days when individuals administered higher-than-average numbers of injections. The observed mean number of daily bolus injections administered across the study population was lower than the optimal number required to reach glycaemic targets (4.8 injections vs. 6-8 injections). Smart pen engagement was significantly associated with improved TIR., Conclusions: Glycaemic control was associated with daily bolus insulin injection frequency and smart pen engagement. A treatment regimen combining an optimal bolus injection strategy, and effective smart pen engagement, may improve glycaemic control among adults with T1D., (© 2023 Novo Nordisk A/S and The Authors. Diabetes, Obesity and Metabolism published by John Wiley & Sons Ltd.)

Osteopontin (OPN) is a bioactive integrin-binding protein found in high concentrations in milk, where it is present both as a full-length protein and as several N-terminally derived fragments. OPN resists gastric digestion, and via interaction with receptors in the gut or by crossing the intestinal barrier into circulation, ingested milk OPN may influence physiological processes. The aim of this study was to investigate OPN interaction with intestinal cells and its transport across models of the intestinal barrier. Immunodetection of OPN incubated with Caco-2 cells at 4 °C and 37 °C showed that OPN binds to the intestinal cells, but it is not internalised. Transepithelial transport was studied using mono- and co-cultures of Caco-2 cells and mucus-producing HT29-MTX cells in transwell membranes. OPN was shown to cross the barrier models in a time-, temperature-, and energy-dependent process inhibited by wortmannin, indicating that the transport takes place via the transcytosis pathway. Analyses of the naturally occurring milk mixture of full-length and N-terminal fragments showed that the N-terminal fragments of OPN bound intestinal cells most effectively and that the fragments were transported across the intestinal membrane models. This suggests that proteolytic processing of OPN increases its biological activity after ingestion.

Background: Patient support apps have risen in popularity and provide novel opportunities for self-management of diabetes. Such apps offer patients to play an active role in monitoring their condition, thereby increasing their own treatment responsibility. Although many health apps require active user engagement to be effective, there is little evidence exploring engagement with mobile health (mHealth)., Objective: This study aims to analyze the extent to which users engage with mHealth for diabetes and identify patient characteristics that are associated with engagement., Methods: The analysis is based on real-world data obtained by Novo Nordisk's Cornerstones4Care Powered by Glooko diabetes support app. User engagement was assessed as the number of active days and using measures expressing the persistence, longevity, and regularity of interaction within the first 180 days of use. Beta regressions were estimated to assess the associations between user characteristics and engagement outcomes for each module of the app., Results: A total of 9051 individuals initiated use after registration and could be observed for 180 days. Among these, 55.39% (5013/9051) used the app for one specific purpose. The average user activity ratio varied from 0.05 (medication and food) to 0.55 (continuous glucose monitoring), depending on the module of the app. Average user engagement was lower if modules required manual data entries, although the initial uptake was higher for these modules. Regression analyses further revealed that although more women used the app (2075/3649, 56.86%), they engaged significantly less with it. Older people and users who were recently diagnosed tended to use the app more actively., Conclusions: Strategies to increase or sustain the use of apps and availability of health data may target the mode of data collection and content design and should take into account privacy concerns of the users at the same time. Users' engagement was determined by various user characteristics, indicating that particular patient groups should be targeted or assisted when integrating apps into the self-management of their disease., (©Anna-Katharina Böhm, Morten Lind Jensen, Mads Reinholdt Sørensen, Tom Stargardt. Originally published in JMIR mHealth and uHealth (http://mhealth.jmir.org), 06.11.2020.)

The aim of this study was to analyse concordance between Danish adults' recorded diet quality and their own assessment of the healthiness and to examine socio-demographic, health and behavioural characteristics associated with an optimistic or pessimistic self-assessment. Data were derived from The Danish National Survey of Diet and Physical Activity 2011-2013 and included a random sample of 3014 adults (18-75 y). Diet quality was evaluated on the basis of seven-day pre-coded food diaries and categorised 'unhealthy', 'somewhat healthy' and 'healthy'. Self-assessment of the healthiness of own diets was registered via personal interviews and categorised healthy enough 'to a high degree', 'to some degree' or 'not at all/only partly'. Highly and somewhat optimistic self-assessment, respectively, were defined as assessing own diets as healthy enough to a high degree or to some degree while having unhealthy diets. Highly and somewhat pessimistic self-assessment, respectively, were defined as assessing own diets as not healthy enough or healthy enough to some degree while having healthy diets. Multiple logistic regression models were used to examine characteristics associated with optimistic and pessimistic self-assessments, respectively. Among individuals with unhealthy diets, 13% were highly optimistic and 42% somewhat optimistic about the healthiness of their diets. Among individuals with healthy diets, 14% were highly pessimistic and 51% somewhat pessimistic about the healthiness of their diets. Highly optimistic self-assessment was associated with increasing age, excellent self-rated health, normal weight and a moderate activity level. Highly pessimistic self-assessment was associated with decreasing age, good self-rated health and being obese. The findings indicate that people seem to use personal health characteristics as important references when assessing the healthiness of their diets., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The need for typing of the swine leukocyte antigen (SLA) is increasing with the expanded use of pigs as models for human diseases and organ-transplantation experiments, their use in infection studies, and for design of veterinary vaccines. Knowledge of SLA sequences is furthermore a prerequisite for the prediction of epitope binding in pigs. The low number of known SLA class I alleles and the limited knowledge of their prevalence in different pig breeds emphasizes the need for efficient SLA typing methods. This study utilizes an SLA class I-typing method based on next-generation sequencing of barcoded PCR amplicons. The amplicons were generated with universal primers and predicted to resolve 68-88% of all known SLA class I alleles dependent on amplicon size. We analyzed the SLA profiles of 72 pigs from four different pig populations; Göttingen minipigs and Belgian, Kenyan, and Danish fattening pigs. We identified 67 alleles, nine previously described haplotypes and 15 novel haplotypes. The highest variation in SLA class I profiles was observed in the Danish pigs and the lowest among the Göttingen minipig population, which also have the highest percentage of homozygote individuals. Highlighting the fact that there are still numerous unknown SLA class I alleles to be discovered, a total of 12 novel SLA class I alleles were identified. Overall, we present new information about known and novel alleles and haplotypes and their prevalence in the tested pig populations.

The quantitative detection of viable pathogen load is an important tool in determining the degree of infection in animals and contamination of foodstuffs. Current conventional culture methods are limited in their ability to determine these levels in Mycobacterium avium subspecies paratuberculosis (MAP) due to slow growth, clumping and low recoverability issues. The principle goal of this study was to evaluate a novel culturing process (TiKa) with unique ability to stimulate MAP growth from low sample loads and dilutions. We demonstrate it was able to stimulate a mean 29-fold increase in recoverability and an improved sensitivity of up to three logs when compared with conventional culture. Using TiKa culture, MAP clumping was minimal and produced visible colonies in half the time required by standard culture methods. Parallel quantitative evaluation of the TiKa culture approach and qPCR on MAP loads in tissue and gut mucosal samples from a MAP vaccine-challenge study, showed good correlations between colony counts (cfu) and qPCR derived genome equivalents (Geq) over a large range of loads with a 30% greater sensitivity for TiKa culture approach at low loads (two logs). Furthermore, the relative fold changes in Geq and cfu from the TiKa culture approach suggests that non-mucosal tissue loads from MAP infected animals contained a reduced proportion of non-viable MAP (mean 19-fold) which was reduced significantly further (mean 190-fold) in vaccinated "reactor" calves. This study shows TiKa culture equates well with qPCR and provides important evidence that accuracy in estimating viable MAP load using DNA tests alone may vary significantly between samples of mucosal and lymphatic origin.

It has been suggested that optimistic self-assessments of unhealthy diets constitute a barrier to the promotion of healthier eating practices. In order to discuss possible reasons for such optimistic assessments, knowledge about the considerations underlying self-assessments of unhealthy diets is needed. The aim of this qualitative study is to explore this issue by comparing considerations underlying the assessments of people who overestimate the healthiness of their unhealthy diets with those of people who express more realistic assessments. Interviewees were recruited among those respondents to the Danish National Survey of Diet and Physical Activity 2011-2013, who had been accorded a low diet index score. A thematic analysis of qualitative interviews is undertaken (N = 16). When interviewees are asked to assess the healthiness of their diets, they draw upon their nutritional knowledge and their perceptions of healthy eating practices. However, these considerations tend to be overruled by more decisive criteria. Thus, diets are assessed as being not exactly healthy, but nevertheless healthy enough - so long as interviewees feel good. Moreover, a personal history of weight status and weight concerns emerge as decisive criteria in self-assessments. Those who experience problems in these areas tend to be realistic about the unhealthy character of their diets, while optimistic assessments appear to be linked to tendencies to perceive oneself as not being overweight, not having experienced weight gain or loss, or not being concerned about weight. This study concludes that decisive criteria in lay people's self-assessments of unhealthy diets - with regard to feeling and looking good - differ markedly from the criteria employed in food-based dietary guidelines. These broader criteria of assessment should be recognized by professionals engaged in planning health promotion strategies with reference to dietary health., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: An ankle-foot orthosis is suggested to actively assist dorsiflexion of the foot by induction of a positive proprioceptive impact to ankle dorsiflexor muscles. However, an objective proof is missing., Objectives: To assess the proprioceptive impact of an ankle-foot orthosis application by Hoffmann reflex recordings of the soleus muscle under static conditions. It was hypothesized that the use of an ankle-foot orthosis facilitated dorsiflexor motor function and thus a decreased the soleus Hoffmann reflex., Study Design: Experimental study in healthy volunteers, pre-post test design., Methods: In all, 20 healthy volunteers were enrolled in order to assess the proprioceptive impact of orthosis application. The Hoffmann reflex was recorded before, during, and after orthosis application., Results: Under orthosis application, the Hoffmann reflex significantly decreased as compared to before (p < 0.05) and after application (p < 0.05)., Conclusions: Findings indicate an inhibition of plantarflexors probably induced by facilitation of ankle dorsiflexors under static conditions. At first glance, it seems that foot orthoses primarily have a stabilizing effect on ankle joints in terms of simple mechanical bandages. However, the present results suggest an additional active impact on proprioceptive control., Clinical Relevance: The putative neuromodulatory effect on motor control may support the application of such ankle-foot orthoses in, for example, drop foot. Furthermore, the objective assessment of a neurophysiological mode of action of orthoses by Hoffmann reflex recordings might be an appropriate primary outcome parameter in clinical trials., (© The International Society for Prosthetics and Orthotics 2014.)

Immunotherapy has increased overall survival of metastatic cancer patients, and cancer antigens are promising vaccine targets. To fulfill the promise, appropriate tailoring of the vaccine formulations to mount in vivo cytotoxic T cell (CTL) responses toward co-delivered cancer antigens is essential. Previous development of therapeutic cancer vaccines has largely been based on studies in mice, and the majority of these candidate vaccines failed to induce therapeutic responses in the subsequent human clinical trials. Given that antigen dose and vaccine volume in pigs are translatable to humans and the porcine immunome is closer related to the human counterpart, we here introduce pigs as a supplementary large animal model for human cancer vaccine development. IDO and RhoC, both important in human cancer development and progression, were used as vaccine targets and 12 pigs were immunized with overlapping 20mer peptides spanning the entire porcine IDO and RhoC sequences formulated in CTL-inducing adjuvants: CAF09, CASAC, Montanide ISA 51 VG, or PBS. Taking advantage of recombinant swine MHC class I molecules (SLAs), the peptide-SLA complex stability was measured for 198 IDO- or RhoC-derived 9-11mer peptides predicted to bind to SLA-1(*)04:01, -1(*)07:02, -2(*)04:01, -2(*)05:02, and/or -3(*)04:01. This identified 89 stable (t½ ≥ 0.5 h) peptide-SLA complexes. By IFN-γ release in PBMC cultures we monitored the vaccine-induced peptide-specific CTL responses, and found responses to both IDO- and RhoC-derived peptides across all groups with no adjuvant being superior. These findings support the further use of pigs as a large animal model for vaccine development against human cancer.

Aims: To examine the development in pedometer-determined physical activity from 2007-2008 to 2011-2012 in the adult Danish population., Methods: The study population comprised two random samples of 18-75-year-old individuals who took part in cross-sectional studies in 2007-2008 (n=224) and 2011-2012 (n=1515). Pedometer data (sealed Yamax SW 200) were obtained for seven consecutive days. Data for 1624 participants (48.2% men) were included in the analysis. An overall step-defined activity level was examined based on a graduated step index (sedentary, low active, somewhat active, active, highly active). The pedometer-determined outcomes were analysed using regression models., Results: A borderline significant decline (p=0.077) from 8788 to 8341 steps/day (-446 (95% confidence intervals -50, 943)) was found between 2007-2008 and 2011-2012. Furthermore, a 23.7% (95% confidence intervals -41.7%, -0.1%) lower overall step-defined activity level was observed in 2011-2012 compared to 2007-2008. These changes were primarily due to a reduced level of activity among women. The proportion of individuals taking ⩾10,000 steps/day decreased non-significantly from 34.8% to 29.3%, whereas the proportion taking <5000 steps/day did not differ between survey periods., Conclusions: This nationally representative survey suggests an overall reduction in the physical activity level among Danish adults. The reduction was due to a shift in the population distribution from higher to lower levels of activity. If this shift is true, it is worrying from a public health perspective. Our study result needs, however, to be confirmed by other population studies., (© 2015 the Nordic Societies of Public Health.)

Aims: To examine social disparities in dietary-, alcohol- and leisure-time physical activity habits in Danish adults from 1995 to 2008 and to test the hypothesis of widening disparities., Methods: Four cross-sectional surveys of representative samples of the Danish population aged 20-75 years were analysed, a total of 7900 respondents. Dietary and alcohol habits were measured by a 7-day pre-coded food diary and physical activity and education by a face-to-face interview. Multivariate analyses were used to test associations between explanatory variables and response variables and the hypothesis of widening disparities., Results: In all analyses, both diet quality and physical inactivity differed systematically with educational group and with healthier habits for the group with long education. Only for alcohol habits was an opposite social gradient seen. Disparities persisted over all time periods. The analyses using "The Concentration Index" did not change any of the conclusions. Gender also influenced diet quality. Men had dietary and alcohol habits furthest away from the guidelines. A trend of healthier habits was also found over the time period., Conclusions: Social disparities in diet quality and leisure-time physical activity were seen for all examined time periods and with no significant change therefore, there is no support to the hypothesis of widening social disparities in all educational groups some improvement of dietary habits was found, especially for those foods where comprehensive initiatives had taken place however, social disparities still exist there is an ongoing need to address these disparities in order to reduce social inequality in health., (© 2014 the Nordic Societies of Public Health.)

Background/objectives: To identify and describe dietary patterns in Danish adults and to examine which demographic and health-related lifestyle factors are associated with dietary patterns., Subjects/methods: Data derived from the Danish national survey of diet and physical activity collected in 2003-2008 and included 1569 men and 1785 women. Diet was assessed by a 7-day pre-coded food diary. Information on age, gender, weight, height, physical activity, smoking habits, educational level and attitudes towards healthy eating habits was derived from face-to-face interviews. Principal component analysis was applied to explore dietary patterns. Associations with lifestyle factors were examined by means of multiple regression analyses., Results: Three major dietary patterns were identified: a 'traditional' pattern correlated with intake of rye bread, white bread, fat on bread, cheese, jam, cold meat, minced meat, potatoes and gravy, and cake and biscuits; a 'health-conscious' pattern correlated with coarse bread, fruit, vegetables, low-fat dairy, nuts, water and tea; and a 'fast food' pattern correlated with pizza, hamburger/spring rolls, crisps, rice and pasta, sugar-sweetened soft drinks and sweets. The 'traditional' pattern was positively associated with male gender and age, whereas the 'health-conscious' pattern was positively associated with being female, increasing age and educational level. The 'fast food' pattern was inversely associated with age and smoking., Conclusions: Three distinct dietary patterns were identified, and associated lifestyle and demographic factors were characterised. The findings are valuable in targeting future nutrition education and will enable more focused strategies in communicating food-based dietary guidelines.

It is generally accepted that CD8 T cells play a major role in tumor control, yet vaccination aimed at eliciting potent CD8 T cell responses are rarely efficient in clinical trials. To try and understand why this is so, we have generated potent adenoviral vectors encoding the endogenous tumor Ags (TA) tyrosinase-related protein-2 (TRP-2) and glycoprotein 100 (GP100) tethered to the invariant chain (Ii). Using these vectors, we sought to characterize the self-TA-specific CD8 T cell response and compare it to that induced against non-self-Ags expressed from a similar vector platform. Prophylactic vaccination with adenoviral vectors expressing either TRP-2 (Ad-Ii-TRP-2) or GP100 (Ad-Ii-GP100) had little or no effect on the growth of s.c. B16 melanomas, and only Ad-Ii-TRP-2 was able to induce a marginal reduction of B16 lung metastasis. In contrast, vaccination with a similar vector construct expressing a foreign (viral) TA induced efficient tumor control. Analyzing the self-TA-specific CD8 T cells, we observed that these could be activated to produce IFN-γ and TNF-α. In addition, surface expression of phenotypic markers and inhibitory receptors, as well as in vivo cytotoxicity and degranulation capacity matched that of non-self-Ag-specific CD8 T cells. However, the CD8 T cells specific for self-TAs had a lower functional avidity, and this impacted on their in vivo performance. On the basis of these results and a low expression of the targeted TA epitopes on the tumor cells, we suggest that low avidity of the self-TA-specific CD8 T cells may represent a major obstacle for efficient immunotherapy of cancer.

Adequate information about physical activity habits is essential for surveillance, implementing, and evaluating public health initiatives in this area. Previous studies have shown that question order and differences in wording result in systematic differences in people's responses to questionnaires; however, this has never been shown for physical activity questions. The aim was to study the influence of different formulations and question order on self-report physical activity in a population-based health interview survey. Four samples of each 1000 adults were drawn at random from the National Person Register. A new question about physical activity was included with minor differences in formulations in samples 1-3. Furthermore, the question in sample 2 was included in sample 4 but was placed in the end of the questionnaire. The mean time spent on moderate physical activity varied between the four samples from 57 to 100 min/day. Question order was associated with the reported number of minutes spent on moderate-intensity physical activity and with prevalence of meeting the recommendation, whereas physical inactivity was associated with the differences in formulation of the question. Questionnaire context influences the way people respond to questions about physical activity significantly and should be tested systematically in validation studies of physical activity questionnaires., (© 2012 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Background: Data on dietary intake and physical activity has been collected from a representative sample of the Danish population from 2003-2008., Objectives: The aim of the present study was to describe the habitual diet in Denmark and to evaluate the overall diet quality using a diet quality index based on the National Food-Based Dietary Guidelines (FBDG), which consists of seven guidelines regarding diet and one regarding physical activity., Design: Data from the Danish National Survey of Diet and Physical Activity 2003-2008 (n=3354) were included. The diet quality index was constructed based on five of the seven dietary guidelines. Individuals were categorised according to quartiles of the diet quality index, and food and nutrient intakes were estimated in each of the groups., Results: Macronutrient distribution did not meet recommendations in any of the groups, as energy from total fat and especially saturated fat was too high. A high intake of high-fat milk products, fat on bread and processed meat contributed to a high intake of total fat and saturated fat, and sugar-sweetened soft drinks contributed to a high intake of added sugars in the group below the lowest quartile of the diet quality index. Individuals above in the highest quartile had higher intakes of 'healthy foods' such as fish, fruit and vegetables, rye bread, and also a higher consumption of water and wine. Overall, intakes of micronutrients were sufficient in all groups., Conclusions: The diet quality index is a useful tool in assessing food and nutrient intake in individuals with high vs. low degree of compliance towards the dietary guidelines, and provides a valuable tool in future studies investigating variations in dietary intakes with respect to lifestyle, demographic and regional differences in Denmark.

Absorption of drugs from subcutaneous tissue depends on several factors, including tissue perfusion at the administration site. Tissue perfusion can be manipulated by e.g. application of local heat. This may subsequently alter the rate or amount of absorption of drugs from a subcutaneous depot. The aim of the present study was to investigate if increased tissue perfusion after controlled local heating can change the absorption of subcutaneously administered short-acting insulin (Actrapid®, 100IU/ml). Thirteen healthy Caucasian males participated in two randomized experimental sessions; one session with locally applied controlled heat at the injection site, and a control session without local heat application. Tissue perfusion (blood flow) was monitored with Laser Doppler Imaging, and blood samples were taken to assess the levels of glucose and insulin. Local heat application at the site of insulin injection significantly enhanced tissue perfusion by approximately 145%. However, no correlation was found between insulin absorption and tissue perfusion. Based on our findings, it was concluded that tissue perfusion is not the rate-limiting factor in the absorption of high-concentration short-acting insulin from a subcutaneous depot. It is suggested that dissociation of insulin hexamers into dimers and monomers is a major rate limiting factor to the absorption.

Intracerebral inoculation of immunocompetent mice with lymphocytic choriomeningitis virus (LCMV) normally results in fatal CD8+ T cell mediated meningoencephalitis. However, in CXCL10-deficient mice, the virus-induced CD8+ T cell accumulation in the neural parenchyma is impaired, and only 30-50% of the mice succumb to the infection. Similar results are obtained in mice deficient in the matching chemokine receptor, CXCR3. Together, these findings point to a key role for CXCL10 in regulating the severity of the LCMV-induced inflammatory process. For this reason, we now address the mechanisms regulating the expression of CXCL10 in the CNS of LCMV-infected mice. Using mice deficient in type I IFN receptor, type II IFN receptor, or type II IFN, as well as bone marrow chimeras expressing CXCL10 only in resident cells or only in bone marrow-derived cells, we analyzed the up-stream regulation as well as the cellular source of CXCL10. We found that expression of CXCL10 initially depends on signaling through the type I IFN receptor, while late expression and up-regulation requires type II IFN produced by the recruited CD8+ T cells. Throughout the infection, the producers of CXCL10 are exclusively resident cells of the CNS, and astrocytes are the dominant expressors in the neural parenchyma, not microglial cells or recruited bone marrow-derived cell types. These results are consistent with a model suggesting a bidirectional interplay between resident cells of the CNS and the recruited virus-specific T cells with astrocytes as active participants in the local antiviral host response.

We previously reported that the lack of serglycin proteoglycan affects secretory granule morphology and granzyme B (GrB) storage in in vitro generated CTLs. In this study, the role of serglycin during viral infection was studied by infecting wild-type (wt) mice and serglycin-deficient (SG(-/-)) mice with lymphocytic choriomeningitis virus (LCMV). Wt and SG(-/-) mice cleared 10(3) PFU of highly invasive LCMV with the same kinetics, and the CD8(+) T lymphocytes from wt and SG(-/-) animals did not differ in GrB, perforin, IFN-gamma, or TNF-alpha content. However, when a less invasive LCMV strain was used, SG(-/-) GrB(+) CD8(+) T cells contained approximately 30% less GrB than wt GrB(+) CD8(+) T cells. Interestingly, the contraction of the antiviral CD8(+) T cell response to highly invasive LCMV was markedly delayed in SG(-/-) mice, and a delayed contraction of the virus-specific CD8(+) T cell response was also seen after infection with vesicular stomatitis virus. BrdU labeling of cells in vivo revealed that the delayed contraction was associated with sustained proliferation of Ag-specific CD8(+) T cells in SG(-/-) mice. Moreover, wt LCMV-specific CD8(+) T cells from TCR318 transgenic mice expanded much more extensively in virus-infected SG(-/-) mice than in matched wt mice, indicating that the delayed contraction represents a T cell extrinsic phenomenon. In summary, the present report points to a novel, previously unrecognized role for serglycin proteoglycan in regulating the kinetics of antiviral CD8(+) T cell responses.

Raising an efficient and sustained immune response to a growing tumour is extremely challenging, as tumours not only lack the capacity to induce an environment optimal for induction of the relevant immune response, but also tend to promote the development of very efficient immunosuppressive mechanisms. This review aims to evaluate selected cancer vaccination approaches using virus-based cancer vaccines. These seem promising based on their capacity to mimic natural infection and hence to efficiently trigger the innate immune system and in turn a potent cellular immune response towards the tumours. However, even when a potent immune response has been induced, this is often not sufficient to eliminate the tumour completely before the cancer cells have had time to evolve new escape mechanisms as a result of the selection pressure from the initial immune response directed towards them. Therefore, it is very likely that it is necessary to combine a therapeutic tumour vaccine with immunomodulating strategies in order to accomplish effective tumour degradation or at least to hinder metastasis. Some of the immunosuppressive mechanisms worth trying to manipulate will be discussed in this review.

Intracerebral (i.c.) infection with lymphocytic choriomeningitis virus (LCMV) is one of the most studied models for virus-induced immunopathology, and based on results from perforin-deficient mice, it is currently assumed that fatal disease directly reflects perforin-mediated cell lysis. However, recent studies have revealed additional functional defects within the effector T cells of LCMV-infected perforin-deficient mice, raising the possibility that perforin may not be directly involved in mediating lethal disease. For this reason, we decided to reevaluate the role of perforin in determining the outcome of i.c. infection with LCMV. We confirmed that the expansion of virus-specific CD8(+) T cells is unimpaired in perforin-deficient mice. However, despite the fact that the virus-specific CD8(+) effector T cells in perforin-deficient mice are broadly impaired in their effector function, these mice invariably succumb to i.c. infection with LCMV strain Armstrong, although a few days later than matched wild-type mice. Upon further investigation, we found that this delay correlates with the delayed recruitment of inflammatory cells to the central nervous system (CNS). However, CD8(+) effector T cells were not kept from the CNS by sequestering in infected extraneural organ sites such as liver or lungs. Thus, the observed dysfunctionality regarding the production of proinflammatory mediators probably results in the delayed recruitment of effector cells to the CNS, and this appears to be the main explanation for the delayed onset of fatal disease in perforin-deficient mice. However, once accumulated in the CNS, virus-specific CD8(+) T cells can induce fatal CNS pathology despite the absence of perforin-mediated lysis and reduced capacity to produce several key cytokines.

CD1 molecules play an important role in the immune system, presenting lipid-containing antigens to T and NKT cells. CD1 genes have long been thought to be as ancient as MHC class I and II genes, based on various arguments, but thus far they have been described only in mammals. Here we describe two CD1 genes in chickens, demonstrating that the CD1 system was present in the last common ancestor of mammals and birds at least 300 million years ago. In phylogenetic analysis, these sequences cluster with CD1 sequences from other species but are not obviously like any particular CD1 isotype. Sequence analysis suggests that the expressed proteins bind hydrophobic molecules and are recycled through intracellular vesicles. RNA expression is strong in lymphoid tissues but weaker to undetectable in some nonlymphoid tissues. Flow cytometry confirms expression from one gene on B cells. Based on Southern blotting and cloning, only two such CD1 genes are detected, located approximately 800 nucleotides apart and in the same transcriptional orientation. The sequence of one gene is nearly identical in six chicken lines. By mapping with a backcross family, this gene could not be separated from the chicken MHC on chromosome 16. Mining the draft chicken genome sequence shows that chicken has only these two CD1 genes located approximately 50 kb from the classical class I genes. The unexpected location of these genes in the chicken MHC suggests the CD1 system was present in the primordial MHC and is thus approximately 600 million years old.

We present atomistic simulations of crystal growth where realistic experimental deposition rates are reproduced, without needing any a priori information on the relevant diffusion processes. Using the temperature accelerated dynamics method, we simulate the deposition of 4 monolayers (ML) of Ag/Ag(100) at the rate of 0.075 ML/s, thus obtaining a boost of several orders of magnitude with respect to ordinary molecular dynamics. In the temperature range analyzed (0-70 K), steering and activated mechanisms compete in determining the surface roughness.

A case of a large pyogenic liver abscess in a 16-year-old boy with appendicitis is presented. The clinical picture, bacteriology and treatment of the disease are discussed on the basis of 397 cases reported in the literature. It is concluded that ultrasound scanning of the liver is essential for early diagnosis and that the treatment of choice is percutaneous needle aspiration in combination with bactericidal antibiotics.