Sarcopenia and sarcopenic obesity among older adults in the nordic countries: a scoping review.

BACKGROUND: Sarcopenia and sarcopenic obesity (SO) are age-related syndromes that may compromise physical and mental health among older adults. The Nordic countries differ from other regions on prevalence of disease, life-style behavior, and life expectancy, which may impact prevalence of sarcopenia and SO. Therefore, the aim of this study is to review the available evidence and gaps within this field in the Nordic countries. METHODS: PubMed, Embase, and Web of science (WOS) were searched up to February 2023. In addition, grey literature and reference lists of included studies were searched. Two independent researcher assessed papers and extracted data. RESULTS: Thirty-three studies out of 6,363 searched studies were included in this scoping review. Overall prevalence of sarcopenia varied from 0.9 to 58.5%. A wide prevalence range was still present for community-dwelling older adults when definition criteria and setting were considered. The prevalence of SO ranged from 4 to 11%, according to the only study on this field. Based on the included studies, potential risk factors for sarcopenia include malnutrition, low physical activity, specific diseases (e.g., diabetes), inflammation, polypharmacy, and aging, whereas increased levels of physical activity and improved dietary intake may reduce the risk of sarcopenia. The few available interventions for sarcopenia were mainly focused on resistance training with/without nutritional supplements (e.g., protein, vitamin D). CONCLUSION: The findings of our study revealed inadequate research on SO but an increasing trend in the number of studies on sarcopenia. However, most of the included studies had descriptive cross-sectional design, small sample size, and applied different diagnostic criteria. Therefore, larger well-designed cohort studies that adhere to uniform recent guidelines are required to capture a full picture of these two age-related medical conditions in Nordic countries, and plan for prevention/treatment accordingly.

Introducing Point-of-Care PCR technology in general practice: Ambiguities, experiences, and perceptions among health care professionals.

In this paper we present findings from a qualitative ethnographic study investigating the experiences and perceptions of general practitioners and other practice staff when introducing a new point of care diagnostic test technology (point of care polymerase chain reaction (POC PCR)) in general practice in Denmark. The ethnographic study was conducted in five general practice clinics, involving observations in four of the clinics and interviews with general practitioners and practice staff in all five clinics. Following an initial analytic phase in which barriers and facilitators in the implementation process of the Point-of-Care test were identified, we developed theoretically informed themes, drawing upon Hartmut Rosa's social theory of technological acceleration. These themes included ambiguous experiences and perceptions of: (i) diagnostic specification and inflation embedded in diagnostic practices; (ii) empowerment and erosion of professional judgment; (iii) strategies of security and insecurity in communication; (iv) the interdependence between professional autonomy and economic structures associated with organizational power; and (v) subjective and organizational time. We discuss how diagnostic technologies simultaneously contribute to and disrupt treatment safety, efficiency, and medical decision-making. Using Rosa's sociological concepts of alienation and resonance, this article furthermore explores how these ambiguous dynamics are experienced in general practice settings. It also examines the implications of navigating a heterogeneous socio-technical and medical landscape and what it means to be a health professional in a contemporary general practice environment that is increasingly shaped by diagnostic technologies.

Designing a tool ensuring older patients the right medication at the right time after discharge from hospital- the first step in a participatory design process.

BACKGROUND: On average, older patients use five or more medications daily, increasing the risk of adverse drug reactions, interactions, or medication errors. Healthcare sector transitions increase the risk of information loss, misunderstandings, unclear treatment responsibilities, and medication errors. Therefore, it is crucial to identify possible solutions to decrease these risks. Patients, relatives, and healthcare professionals were asked to design the solution they need. METHODS: We conducted a participatory design approach to collect information from patients, relatives, and healthcare professionals. The informants were asked to design their take on a tool ensuring that patients received the correct medication after discharge from the hospital. We included two patients using five or more medications daily, one relative, three general practitioners, four nurses from different healthcare sectors, two hospital physicians, and three pharmacists. RESULTS: The patients' solution was a physical location providing a medication overview, including side effects and interactions. Healthcare professionals suggested different solutions, including targeted and timely information that provided an overview of the patient's diagnoses, treatment and medication. The common themes identified across all sub-groups were: (1) Overview of medications, side effects, and diagnoses, (2) Sharing knowledge among healthcare professionals, (3) Timely discharge letters, (4) Does the shared medication record and existing communication platforms provide relevant information to the patient or healthcare professional? CONCLUSION: All study participants describe the need for a more concise, relevant overview of information. This study describes elements for further elaboration in future participatory design processes aimed at creating a tool to ensure older patients receive the correct medication at the correct time.

Increasing melanoma incidence with unchanged mortality: more sunshine, better treatment, increased diagnostic activity, overdiagnosis or lowered diagnostic threshold?

BACKGROUND: Increasing melanoma incidence with less increasing mortality is observed in several countries. This discrepancy is not well understood. OBJECTIVE: In this study, our aim was to discuss factors (UV exposure, melanoma treatment, diagnostic activity, overdiagnosis, pathologists' diagnostic threshold and clinicians' propensity to remove suspect skin lesions) that may influence melanoma incidence and mortality in Denmark. METHODS: This was a register study with the number of melanocyte-related lesions and melanoma mortality based on comprehensive national pathology and mortality databases for the period 1999-2019. We investigated melanocyte-related diagnoses and mortality in a population of 5.5 million with national health care system. Age adjusted melanoma mortality and age-adjusted incidence of benign nevi, atypical lesion, or melanoma-in-situ and of invasive melanoma were computed for data analysis. RESULTS: In total 1,434,798 biopsies were taken from 704,682 individuals (65% female). Mean age at biopsy was 39.8 years in men and 37.6 in women. In men and women, the incidence of invasive melanoma increased by 87% during the period 1999-2011. During the subsequent period it increased by 9% in men but remained unchanged in women. The incidence of melanoma in-situ increased by 476% in men and 357% in women during the study period, while the increases for atypical melanocytic lesions were 1928% and 1686%, respectively. Biopsy rates increased by 153% in men and 118% in women from 1999 through 2011 but fell by 20% in men and 22% in women during the subsequent period. Mortality varied slightly from year to year without any significant time trend for men or women.We identified no evidence of increased UV exposure over the latest 30 years in Denmark. Immunotherapy of advanced melanoma was introduced in Denmark in 2010 and came in general use in 2014. CONCLUSIONS: Comprehensive national data demonstrate increasing melanoma incidence correlated with increasing biopsy rates, but with no change in mortality. Previously suggested explanations for such a trend are lowered threshold of melanoma diagnosis among pathologists, increased diagnostic activity in the presence of overdiagnosis and improved melanoma treatment. Because the study is observational and because we have more explanatory factors than outcomes, the findings do not warrant conclusions about causal relationships.

Patients' views on usefulness and effects of a risk communication tool for cardiovascular disease: a qualitative analysis.

BACKGROUND: Failing to comprehend risk communication might contribute to poor treatment adherence. Using hypertension as a case, we investigated how a risk communication tool for patients with an elevated risk of cardiovascular disease was perceived. METHODS: As part of a large project featuring a randomised controlled trial in a general practice setting in the Region of Southern Denmark, we conducted a semi-structured individual interview study. The study included patients with hypertension who had used an intervention comprising a visual and dynamic cardiovascular risk communication tool, along with receiving recurring emails providing advice on a healthy lifestyle. The analyses were based on Malterud's Systematic Text Condensation. RESULTS: This article focuses solely on the results of the interview study, which comprised a total of 9 conducted and analysed interviews. The IT setup had a major impact on adherence to the intervention. A positive impact was found when the IT setup was perceived as easy to use and accessible, while a negative impact was noted when it malfunctioned. The intervention increased patients' self-reported insight into risk of cardiovascular disease. Patients reported the intervention and their risk of cardiovascular disease to become less important to them when they had more severe comorbidities. The involved health professional was very important for treatment adherence when communicating risk visually. Patients expressed trust in their general practitioners, and the general practitioners' attitudes toward the intervention affected patients' perceptions of its usefulness. While the informants reported an increased awareness of their risk of cardiovascular disease, none of them felt more concerned. CONCLUSIONS: Patients reported an increase in their perceived insight into the risk of cardiovascular disease but not an increased concern. Our findings align with previous studies emphasizing the importance of patients' motivation as well as risk perception for adherence. General practitioners have an important role when implementing new tools for patients.

An Albumin-Holliday Junction Biomolecular Modular Design for Programmable Multifunctionality and Prolonged Circulation.

Combinatorial properties such as long-circulation and site- and cell-specific engagement need to be built into the design of advanced drug delivery systems to maximize drug payload efficacy. This work introduces a four-stranded oligonucleotide Holliday Junction (HJ) motif bearing functional moieties covalently conjugated to recombinant human albumin (rHA) to give a "plug-and-play" rHA-HJ multifunctional biomolecular assembly with extended circulation. Electrophoretic gel-shift assays show successful functionalization and purity of the individual high-performance liquid chromatography-purified modules as well as efficient assembly of the rHA-HJ construct. Inclusion of an epidermal growth factor receptor (EGFR)-targeting nanobody module facilitates specific binding to EGFR-expressing cells resulting in approximately 150-fold increased fluorescence intensity determined by flow cytometric analysis compared to assemblies absent of nanobody inclusion. A cellular recycling assay demonstrated retained albumin-neonatal Fc receptor (FcRn) binding affinity and accompanying FcRn-driven cellular recycling. This translated to a 4-fold circulatory half-life extension (2.2 and 0.55 h, for the rHA-HJ and HJ, respectively) in a double transgenic humanized FcRn/albumin mouse. This work introduces a novel biomolecular albumin-nucleic acid construct with extended circulatory half-life and programmable multifunctionality due to its modular design.

Secondary exposure to heavy metal in genetically susceptible mice leads to acceleration of autoimmune response.

Exposure to mercury (Hg) and silver (Ag) has been shown to induce autoimmune diseases in genetically susceptible rodents. Here, A.SW mice were initially exposed to HgCl2, AgNO3 or tap water (control) for 3 weeks. After 13 weeks of stoppage, all mice had secondary exposure to 203HgCl2. After secondary exposure, higher and earlier ANoA titers were observed in mice initially exposed to Hg or Ag compared to control. Further, mice initially exposed to Ag showed higher total IgG1 and IgG2a, Whole Body Retention and lymph nodes and spleen accumulation of Hg compared to mice initially exposed to Hg and controls. These findings showed an earlier and stronger immunological response in A.SW mice compared with control, following re-exposure to heavy metals indicating an immunological memory. Additionally, secondary exposure to a different heavy metal may aggravate the effects of exposure of at least one of the metals indicating cross-reactivity.

The effect of psychological interventions targeting overweight and obesity in school-aged children: a systematic review and meta-analysis.

BACKGROUND: Multi-component psychological interventions may mitigate overweight and obesity in children and adolescents. Evidence is, however, scattered on the effectiveness of such interventions. This study aims to review the available evidence on the effectiveness of multi-component psychological interventions on anthropometric measures of school-aged children with overweight or obesity. METHODS: We systematically searched international databases/search engines including PubMed and NLM Gateway (for MEDLINE), Web of Science, SCOPUS, and Google Scholar up to November 2022 for relevant articles pertaining to psychological weight-loss interventions targeting school-aged children. Two reviewers screened and extracted pertinent data. The quality of included studies was assessed using the Cochrane Risk of Bias Tool for Randomized Trials. Random effect meta-analysis was used to calculate, and pool standardized mean differences (SMD). We distinguished between intervention and maintenance effects. Intervention effects were defined as the mean change in outcome measurement detected between baseline and post-treatment. Maintenance effects were defined as the mean change in outcome measurement between post-treatment and last follow-up. RESULTS: Of 3,196 studies initially identified, 54 and 30 studies were included in the qualitative and quantitative syntheses, respectively. Most studies reported on group-based interventions. The significant effects of intervention on BMI z-score (SMD -0.66, 95% CI: -1.15, -0.17) and WC (SMD -0.53, 95% CI: -1.03, -0.04) were observed for interventions that centered on motivational interviewing and cognitive behavioral therapy, respectively. Mean BMI and WC did not differ significantly between post-treatment and last follow-up measurement (maintenance effect), indicating that an initial weight loss obtained through the intervention period could be maintained over time. CONCLUSIONS: Findings indicate that motivational interviewing and cognitive behavioral therapy as interventions to reduce BMI z-score (generalized obesity) and waist circumference (abdominal obesity) are effective and durable. However, detailed analyses on individual components of the interventions are recommended in future effectiveness studies.

Handling polypharmacy -a qualitative study using focus group interviews with older patients, their relatives, and healthcare professionals.

BACKGROUND: On average, older patients use five or more medications daily. A consequence is an increased risk of adverse drug reactions, interactions, or medication errors. Therefore, it is important to understand the challenges experienced by the patients, relatives, and healthcare professionals pertinent to the concomitant use of many drugs. METHODS: We conducted a qualitative study using focus group interviews to collect information from patients, relatives, and healthcare professionals regarding older patients' management of prescribed medicine. We interviewed seven patients using five or more medications daily, three relatives, three general practitioners, nine nurses from different healthcare sectors, one home care assistant, two hospital physicians, and four pharmacists. RESULTS: The following themes were identified: (1) Unintentional non-adherence, (2) Intentional non-adherence, (3) Generic substitution, (4) Medication lists, (5) Timing and medication schedule, (6) Medication reviews and (7) Dose dispensing/pill organizers. CONCLUSION: Medication is the subject of concern among patients and relatives. They become confused and insecure about information from different actors and the package leaflets. Therefore, patients often request a thorough medication review to provide an overview, knowledge of possible side effects and interactions, and a clarification of the medication's timing. In addition, patients, relatives and nurses all request an indication of when medicine should be taken, including allowable deviations from this timing. Therefore, prescribing physicians should prioritize communicating information regarding these matters when prescribing.

Evaluating the influence of anatomical accuracy and electrode positions on EEG forward solutions.

Generating realistic volume conductor models for forward calculations in electroencephalography (EEG) is not trivial and several factors contribute to the accuracy of such models, two of which are its anatomical accuracy and the accuracy with which electrode positions are known. Here, we investigate effects of anatomical accuracy by comparing forward solutions from SimNIBS, a tool which allows state-of-the-art anatomical modeling, with well-established pipelines in MNE-Python and FieldTrip. We also compare different ways of specifying electrode locations when digitized positions are not available such as transformation of measured positions from standard space and transformation of a manufacturer layout. Substantial effects of anatomical accuracy were seen throughout the entire brain both in terms of field topography and magnitude with SimNIBS generally being more accurate than the pipelines in MNE-Python and FieldTrip. Topographic and magnitude effects were particularly pronounced for MNE-Python which uses a three-layer boundary element method (BEM) model. We attribute these mainly to the coarse representation of the anatomy used in this model, in particular differences in skull and cerebrospinal fluid (CSF). Effects of electrode specification method were evident in occipital and posterior areas when using a transformed manufacturer layout whereas transforming measured positions from standard space generally resulted in smaller errors. We suggest modeling the anatomy of the volume conductor as accurately possible and we hope to facilitate this by making it easy to export simulations from SimNIBS to MNE-Python and FieldTrip for further analysis. Likewise, if digitized electrode positions are not available, a set of measured positions on a standard head template may be preferable to those specified by the manufacturer.

Subjective expectation of reaching age 85: agreement with population statistics and association with behavioral and psychological factors.

BACKGROUND: It is not clearly known how well Danes estimate their chances of reaching the average life expectancy and whether identifiable population subgroups misestimate their life expectancy, and potentially also investments and savings in health and pensions. Therefore, in this study, we examined on the individual level whether subjective life expectancy is in line with the statistically calculated chance of reaching age 85, and further explored the psychological and behavioral factors associated with under or overestimation. METHODS: We opted for a cross-sectional survey design based on a sample of 5,379 Danish citizens aged 50-70 years, returning a web-based questionnaire with socio-demographic data supplemented from a national registry. Average participant estimates of their chance of reaching age 85 for each age range and sex group were compared with actuarial data. We then performed multiple linear regression analyses to examine factors associated with the subjective expectancy of reaching age 85 years. RESULTS: We found that 32% of females and 23% of males reported 100% certainty of reaching age 85, and average expected survival chance exceeded the statistically predicted survival chance for 23% of males and 16% for females in age-ranges 50-60 and 61-70. Our multivariable analysis found that health literacy, internal health locus of control, willingness to take health risks, self-rated health, and health and life satisfaction all showed a significant positive association with expectation of reaching age 85. Moreover, those on daily medications, ex- or current smokers, and heavy drinkers were significantly less optimistic about reaching age 85. CONCLUSIONS: Particularly for the population groups with inaccurate life expectancies, the significant associations with psychological and behavioral factors open a way for initiatives based on behavior change theories to reach a better agreement between subjective and statistical life expectancy.

Antigen footprint governs activation of the B cell receptor.

Antigen binding by B cell receptors (BCR) on cognate B cells elicits a response that eventually leads to production of antibodies. However, it is unclear what the distribution of BCRs is on the naïve B cell and how antigen binding triggers the first step in BCR signaling. Using DNA-PAINT super-resolution microscopy, we find that most BCRs are present as monomers, dimers, or loosely associated clusters on resting B cells, with a nearest-neighbor inter-Fab distance of 20-30 nm. We leverage a Holliday junction nanoscaffold to engineer monodisperse model antigens with precision-controlled affinity and valency, and find that the antigen exerts agonistic effects on the BCR as a function of increasing affinity and avidity. Monovalent macromolecular antigens can activate the BCR at high concentrations, whereas micromolecular antigens cannot, demonstrating that antigen binding does not directly drive activation. Based on this, we propose a BCR activation model determined by the antigen footprint.

Readiness to Accept Genetic Testing for Personalized Medicine: Survey Findings on the Role of Socio-Demographic Characteristics, Health Vulnerabilities, Perceived Genetic Risk and Personality Factors.

Studies from various countries have shown that majorities would accept genetic testing for personalization of treatment, but little is known about differences among population subgroups. The present study investigated whether readiness to accept a hypothetical cost-free offer of genetic testing to personalize treatment depends on socio-demographic characteristics, health-related vulnerabilities, personal dispositions, and prior awareness about personalized medicine. The study was based on a cross-sectional survey design. Out of a representative initial sample of 50-80-year-old Danish citizens (n = 15,072), n = 6807 returned a fully answered web-based questionnaire. Socio-demographic data were added from a national registry. Data were analyzed by multivariable logistic regression. A large majority of respondents (78.3%) expressed their readiness to be tested. Rates were higher in men, younger persons, and those with higher income. Additionally, ex-smokers and obese persons as well as those less satisfied with their health and respondents who perceived a personal genetic risk were more interested, as were those with higher internal health control, higher extraversion, higher emotional stability, and those who had not heard about this option before. Further research should investigate the specific concerns among population subgroups which need being addressed by systematic communication efforts in a clinical but also a broader public health context.

Long-term weight loss in a 24-month primary care-anchored telehealth lifestyle coaching program: Randomized controlled trial.

Long-term weight loss can reduce the risk of type 2 diabetes for people living with obesity and reduce complications for patients diagnosed with type 2 diabetes. We investigated whether a telehealth lifestyle-coaching program (Liva) leads to long-term (24 months) weight loss compared to usual care. In a randomized controlled trial, n = 340 participants living with obesity with or without type 2 diabetes were enrolled and randomized via an automated computer algorithm to an intervention group (n = 200) or to a control group (n = 140). The telehealth lifestyle-coaching program comprised of an initial one-hour face-to-face motivational interview followed by asynchronous telehealth coaching. The behavioural change techniques used were enabled by individual live monitoring. The primary outcome was a change in body weight from baseline to 24 months. Data were assessed for n = 136 participants (40%), n = 81 from the intervention group and n = 55 from the control group, who completed the 24-month follow-up. After 24 months mean body weight and body mass index were reduced significantly for completers in both groups, but almost twice as much was registered for those in the intervention group which was not significant between groups -4.4 (CI -6.1; -2.8) kg versus -2.5 (CI -3.9; -1.1) kg, P = 0.101. Haemoglobin A1c was significantly reduced in the intervention group -3.1 (CI -5.0; -1.2) mmol/mol, but not in the control group -0.2 (CI -2.4; -2.0) mmol/mol without a significant between group difference (P = 0.223). Low completion was partly due to coronavirus disease 2019. Telehealth lifestyle coaching improve long-term weight loss (> 24 months) for obese people with and without type 2 diabetes compared to usual care.

Long-term Weight Loss in a Primary Care-Anchored eHealth Lifestyle Coaching Program: Randomized Controlled Trial.

BACKGROUND: Long-term weight loss in people living with obesity can reduce the risk and progression of noncommunicable diseases. Observational studies suggest that digital coaching can lead to long-term weight loss. OBJECTIVE: We investigated whether an eHealth lifestyle coaching program for people living with obesity with or without type 2 diabetes led to significant, long-term (12-month) weight loss compared to usual care. METHODS: In a randomized controlled trial that took place in 50 municipalities in Denmark, 340 people living with obesity with or without type 2 diabetes were enrolled from April 16, 2018, to April 1, 2019, and randomized via an automated computer algorithm to an intervention (n=200) or a control (n=140) group. Patients were recruited via their general practitioners, the Danish diabetes organization, and social media. The digital coaching intervention consisted of an initial 1-hour face-to-face motivational interview followed by digital coaching using behavioral change techniques enabled by individual live monitoring. The primary outcome was change in body weight from baseline to 12 months. RESULTS: Data were assessed for 200 participants, including 127 from the intervention group and 73 from the control group, who completed 12 months of follow-up. After 12 months, mean body weight and BMI were significantly reduced in both groups but significantly more so in the intervention group than the control group (-4.5 kg, 95% CI -5.6 to -3.4 vs -1.5 kg, 95% CI -2.7 to -0.2, respectively; P<.001; and -1.5 kg/m2, 95% CI -1.9 to -1.2 vs -0.5 kg/m2, 95% CI -0.9 to -0.1, respectively; P<.001). Hemoglobin A1c was significantly reduced in both the intervention (-6.0 mmol/mol, 95% CI -7.7 to -4.3) and control (-4.9 mmol/mol, 95% CI -7.4 to -2.4) groups, without a significant group difference (all P>.46). CONCLUSIONS: Compared to usual care, digital lifestyle coaching can induce significant weight loss for people living with obesity, both with and without type 2 diabetes, after 12 months. TRIAL REGISTRATION: ClinicalTrials.gov NCT03788915; https://clinicaltrials.gov/ct2/show/NCT03788915.

[Treatment of peritoneal metastases from colorectal cancer].

Ten per cent of patients with colorectal cancer will develop peritoneal metastases. These metastases are known to have a lesser response to systemic chemotherapy than liver- and lung metastases. Randomized studies have shown that adding cytoreductive surgery and heated intraperitoneal chemotherapy (HIPEC) to this treatment gives a better chance of survival. Recently, a randomized study failed to show a difference in survival between cytoreductive surgery plus HIPEC versus cytoreductive surgery alone. These facts are summarized in this review.

Reversing Type 2 Diabetes in a Primary Care-Anchored eHealth Lifestyle Coaching Programme in Denmark: A Randomised Controlled Trial.

The goal of this trial was to investigate whether an eHealth lifestyle coaching programme led to significant weight loss and decreased Haemoglobin A1c (HbA1c) in patients with type 2 diabetes. In an RCT, 170 patients were enrolled from 2018 to 2019 for intervention or control. Inclusion criteria were diagnosed with type 2 diabetes, BMI 30−45 kg/m2, and aged 18−70 years. Exclusion criteria were lacks internet access, pregnant or planning a pregnancy, or has a serious disease. Primary and secondary outcomes were a reduction in body weight and HbA1c. At six months, 75 (75%) patients in the intervention group and 53 (76%) patients in the control group remained in the trial. The mean body weight loss was 4.2 kg (95% CI, −5.49; −2.98) in the intervention group and 1.5 kg (95% CI, −2.57; −0.48) in the control group (p = 0.005). In the intervention group, 24 out of 62 patients with elevated HbA1c at baseline (39%) had a normalized HbA1c < 6.5% at six months, compared to 8 out of 40 patients with elevated HbA1c at baseline (20%) in the control group (p = 0.047). The eHealth lifestyle coaching programme can lead to significant weight loss and decreased HbA1c among patients with type 2 diabetes, compared to standard care.

Site-specific nanobody-oligonucleotide conjugation for super-resolution imaging.

Camelid single-domain antibody fragments, also called nanobodies, constitute a class of binders that are small in size (~15 kDa) and possess antigen-binding properties similar to their antibody counterparts. Facile production of recombinant nanobodies in several microorganisms has made this class of binders attractive within the field of molecular imaging. Particularly, their use in super-resolution microscopy has improved the spatial resolution of molecular targets due to a smaller linkage error. In single-molecule localization microscopy techniques, the effective spatial resolution can be further enhanced by site-specific fluorescent labeling of nanobodies owing to a more homogeneous protein-to-fluorophore stoichiometry, reduced background staining and a known distance between dye and epitope. Here, we present a protocol for site-specific bioconjugation of DNA oligonucleotides to three distinct nanobodies expressed with an N- or C-terminal unnatural amino acid, 4-azido-L-phenylalanine (pAzF). Using copper-free click chemistry, the nanobody-oligonucleotide conjugation reactions were efficient and yielded highly pure bioconjugates. Target binding was retained in the bioconjugates, as demonstrated by bio-layer interferometry binding assays and the super-resolution microscopy technique, DNA points accumulation for imaging in nanoscale topography (PAINT). This method for site-specific protein-oligonucleotide conjugation can be further extended for applications within drug delivery and molecular targeting where site-specificity and stoichiometric control are required.

Reasons for participation and non-participation in colorectal cancer screening.

OBJECTIVES: The aim of the present analysis is to identify the reasons for accepting or rejecting the invitation to be screened by the Faecal Immunochemical Test as part of the free Danish screening programme for colorectal cancer (CRC). STUDY DESIGN: A cross-sectional representative survey of 15,072 Danish citizens aged 50-80 years was collected in 2019 via a Web-based questionnaire administered by Statistics Denmark. Among the net sample of 6807 respondents (45%), 177 were excluded because of current treatment for colorectal disease. METHODS: To determine the reasons for accepting or refusing the invitation to be screened for CRC, a latent class analysis was conducted, which allowed participants to provide several reasons for acceptance or rejection of screening. RESULTS: The most important reason for participating in CRC screening was the active public programme. A further reason for participation was the perceived risk for CRC, mainly in combination with the public programme. The reasons for participation did not differ between individuals who had participated and those who intended to participate when offered. Among participants who declined screening, the most frequent reasons were that they forgot to participate or that they were concerned about the unpleasant test procedure. Among individuals who intended to decline screening, a perceived low risk for CRC was the most frequently cited reason. CONCLUSIONS: Recommendation from a general practitioner (GP) was not given as a frequent reason for CRC screening participation which is discussed as a challenge to participation rates in population based screening program The main reasons reported for non-participation in CRC screening (i.e. forgot to participate or the unpleasant test procedure) might be addressed by a stronger endorsement from GPs.

Functionalized Acyclic (l)-Threoninol Nucleic Acid Four-Way Junction with High Stability In Vitro and In Vivo.

Oligonucleotides are increasingly being used as a programmable connection material to assemble molecules and proteins in well-defined structures. For the application of such assemblies for in vivo diagnostics or therapeutics it is crucial that the oligonucleotides form highly stable, non-toxic, and non-immunogenic structures. Only few oligonucleotide derivatives fulfil all of these requirements. Here we report on the application of acyclic l-threoninol nucleic acid (aTNA) to form a four-way junction (4WJ) that is highly stable and enables facile assembly of components for in vivo treatment and imaging. The aTNA 4WJ is serum-stable, shows no non-targeted uptake or cytotoxicity, and invokes no innate immune response. As a proof of concept, we modify the 4WJ with a cancer-targeting and a serum half-life extension moiety and show the effect of these functionalized 4WJs in vitro and in vivo, respectively.