CD38-Specific Biparatopic Heavy Chain Antibodies Display Potent Complement-Dependent Cytotoxicity Against Multiple Myeloma Cells

CD38 is overexpressed by multiple myeloma cells and has emerged as a target for therapeutic antibodies. Nanobodies are soluble single domain antibody fragments derived from the VHH variable domain of heavy chain antibodies naturally occurring in camelids. We previously identified distinct llama nanobodies that recognize three non-overlapping epitopes of the extracellular domain of CD38. Here, we fused these VHH domains to the hinge, CH2, and CH3 domains of human IgG1, yielding highly soluble chimeric llama/human heavy chain antibodies (hcAbs). We analyzed the capacity of these hcAbs to mediate complement-dependent cytotoxicity (CDC) to CD38-expressing human multiple myeloma and Burkitt lymphoma cell lines. Combinations of two hcAbs that recognize distinct, non-overlapping epitopes of CD38 mediated potent CDC, in contrast to the hcAb monotherapy with only weak CDC capacity. Similarly, combining daratumumab with a hcAb that recognizes a non-overlapping epitope resulted in dramatically enhanced CDC. Further, introducing the E345R HexaBody mutation into the CH3 domain strongly enhanced the CDC potency of hcAbs to CD38-expressing cells. Exploiting their high solubility, we genetically fused two distinct nanobodies into heteromeric dimers via a flexible peptide linker and then fused these nanobody dimers to the hinge, CH2 and CH3 domains of human IgG1, yielding highly soluble, biparatopic hcAbs. These biparatopic hcAbs elicited CDC toward CD38-expressing myeloma cells more effectively than daratumumab. Our results underscore the advantage of nanobodies vs. pairs of VH and VL domains for constructing bispecific antibodies. Moreover, the CD38-specific biparatopic heavy chain antibodies described here represent potential new powerful therapeutics for treatment of multiple myeloma

Sex- and age-specific normative values for handgrip strength and components of the Senior Fitness Test in community-dwelling older adults aged 65–75 years in Germany: results from the OUTDOOR ACTIVE study

Background!#!Physical fitness is a key component of independent living and healthy ageing. For the measurement of physical fitness in older adults, the Senior Fitness Test is a commonly used tool. The objective of this study is to calculate sex- and age-specific normative values for handgrip strength and components of the Senior Fitness Test for older adults (65-75 years) in Germany.!##!Methods!#!Cross-sectional data of 1657 community-dwelling older adults residing in Bremen, Germany (53% female) were included in this study. Physical fitness was assessed using the following measurements of the Senior Fitness Test battery: 30s-chair stand test, 2 min-step test, sit-and-reach test, and back scratch test. In addition, handgrip strength was measured using a Saehan DHD-3 digital hand dynamometer SH1003. Sex- and age specific normative values were calculated for the 1st, 3rd, 10th, 25th, 50th, 75th, 90th, 97th, and 99th percentile using the GAMLSS method.!##!Results!#!The normative values show differences dependent on sex and age. For handgrip strength, the 30s-chair stand test and the 2 min-step test, normative values were higher for men, while women reached higher values in the sit-and-reach test and the back scratch test. For both, men and women, normative values declined with age.!##!Conclusions!#!This study provides sex- and age-specific normative values for handgrip strength and components of the Senior Fitness Test for older adults in Germany. They might be useful for future research and for the application in practice

Men’s sheds as community-based health promotion for men aged 50 plus: protocol for a mixed-methods systematic review

Background!#!Men are less likely to participate in health promotion. One approach to reach men is the concept of men's sheds. This community-based health promotion concept brings older men together to engage in joint activities. Prior research revealed various health-related effects of men's sheds, such as benefits for well-being and mental health. To strengthen the current evidence base of men's sheds, a mixed-methods systematic review analysing the association between participation and self-rated health, subjective well-being, and social isolation will be conducted. Moreover, information on how to successfully implement men's sheds will be gathered.!##!Methods!#!This mixed-methods systematic review will follow the guidelines of the Joanna Briggs Institute (JBI). The databases MEDLINE (via PubMed), Scopus, Web of Science, and OpenGrey and the websites of men's sheds associations will be searched for publications. Additionally, a hand search in the reference lists of the included publications will be conducted. Qualitative and quantitative studies published in English, German, or French will be considered for inclusion. The quality of the selected studies will be assessed using the JBI critical appraisal checklists. Following the convergent integrated approach, quantitative data will be transformed into textual descriptions, and subsequently combined with data from qualitative studies as well as from the qualitative components of mixed-methods studies in a simultaneous data synthesis.!##!Discussion!#!The results of this systematic review will lead to a comprehensive understanding of the current evidence base regarding the effectiveness of men's sheds. Furthermore, they will provide useful implications for the implementation of men's sheds.!##!Systematic review registration!#!PROSPERO CRD42020219390

Time allocation to active domains, physical activity, and health indicators in older adults: cross-sectional results from the OUTDOOR ACTIVE study

Background!#!Physical activity (PA) is one of the key determinants of healthy ageing. Research showed that time allocation plays an important role in PA. Therefore, an understanding of the time use of older adults is crucial for developing PA programs. The aim of this study was to examine the associations of time allocation and objectively measured PA, and several health indicators in older adults.!##!Methods!#!In this cross-sectional study all 915 participants of the OUTDOOR ACTIVE study were included. The participants were 65 to 75 years old and resided in a subdistrict of Bremen, Germany (50.9% female). The active domains were derived from the SLOTH model (leisure activities, occupation, active transport, home-based activities). PA was objectively measured with accelerometers over seven consecutive days. Binary logistic regressions were used to test the associations of total PA and time spent in the domains with several health indicators (self-rated health, overweight, obesity, activities of daily living (ADL)).!##!Results!#!Participants over the age of 70 years were significantly less physically active than those under 70 years and women were significantly more physically active than men. Regardless of age and sex, most time was spent on home-based activities (women: 118.5 ± 87.8 min/day; men: 80.2 ± 69.4 min/day). Both PA and time spent on leisure activities were associated with a lower risk of bad self-rated health (0.36; 95%-CL: 0.20, 0.65 for PA; 0.93; 95%-CL: 0.87, 0.99 for leisure activities) and less limitations in ADL. PA and active transport seemed to lower the risk of overweight (0.39; 95%-CL: 0.25, 0.62 for PA; 0.80; 95%-CL: 0.69, 0.93 for active transport) and obesity (0.36; 95%-CL: 0.21, 0.60 for PA; 0.77; 95%-CL: 0.64, 0.92 for active transport). Having an occupation was associated with a lower risk of bad self-rated health (0.60; 95%-CL: 0.40, 0.92).!##!Conclusions!#!The results of this study provide insights in the time allocation to active domains and total PA of older adults, as well as the associations with health indicators. These findings have important implications for the development of PA programs and guidelines. Future research should examine the associations further in longitudinal studies

Accelerometer-assessed outdoor physical activity is associated with meteorological conditions among older adults: Cross-sectional results from the OUTDOOR ACTIVE study.

BACKGROUND:Meteorological conditions are potential determinants of physical activity (PA). A profound understanding of the determinants of PA behaviour is required for PA promotion. This study examined the association between accelerometer-assessed PA and meteorological conditions among older adults. METHODS:This cross-sectional study included data of 577 adults aged 65-75 years living in Bremen, Germany (52% female; 3278 days). PA was measured with accelerometers for seven consecutive days (10/15-08/16). A threshold of 240 lx was used to differentiate between outdoor physical activity (OPA) and indoor physical activity (IPA). Linear mixed models estimated the association between PA (daily accelerometer counts per minute (CPM)) and meteorological factors (temperature, cloud cover, wind, and no precipitation) derived by principal component analysis. RESULTS:The analyses showed associations between PA in CPM and the meteorological factors temperature (93.7; 95%-CL: 64.9, 122.5) and no precipitation (48.4; 95%-CL: 19.8, 77.0) in women and wind (-40.3; 95%-CL: -59.7, -20.8) and no precipitation (30.1; 95%-CL: 5.6, 54.6) in men. After distinguishing in OPA and IPA for a subsample of 128 participants (473 days), the sex differences were no longer present. OPA in CPM was associated with temperature (women: 174.5; 95%-CL: 81.3, 267.6; men: 183.3; 95%-CL: 81.2, 285.4), cloud cover (women: -153.0; 95%-CL: -200.3, -105.7; men: -123.2; 95%-CL: -174.7, -71.7), and wind (women: -118.6; 95%-CL: -189.6; -47.7; men: -96.9; 95%-CL: -177.0, -16.7). No association between OPA and no precipitation was found (women: 2.9; 95%-CL: -89.0, 94.8; men: -17.1; 95%-CL: -116.7, 82.4). CONCLUSIONS:The results of this study emphasize the importance of meteorological conditions as environmental determinants of PA among older adults. Meteorological conditions should be accounted for in the unbiased assessment of habitual PA and the development of PA promotion programs. Future research should focus on the associations of OPA and IPA with meteorological conditions in different climatic regions

Communicating prognosis to women with early breast cancer – overview of prediction tools and the development and pilot testing of a decision aid

Abstract Background Shared decision-making in oncology requires information on individual prognosis. This comprises cancer prognosis as well as competing risks of dying due to age and comorbidities. Decision aids usually do not provide such information on competing risks. We conducted an overview on clinical prediction tools for early breast cancer and developed and pilot-tested a decision aid (DA) addressing individual prognosis using additional chemotherapy in early, hormone receptor-positive breast cancer as an example. Methods Systematic literature search on clinical prediction tools for the effects of drug treatment on survival of breast cancer. The DA was developed following criteria for evidence-based patient information and International Patient Decision Aids Standards. We included data on the influence of age and comorbidities on overall prognosis. The DA was pilot-tested in focus groups. Comprehension was additionally evaluated through an online survey with women in breast cancer self-help groups. Results We identified three prediction tools: Adjuvant!Online, PREDICT and CancerMath. All tools consider age and tumor characteristics. Adjuvant!Online considers comorbidities, CancerMath displays age-dependent non-cancer mortality. Harm due to therapy is not reported. Twenty women participated in focus groups piloting the DA until data saturation was achieved. A total of 102 women consented to participate in the online survey, of which 86 completed the survey. The rate of correct responses was 90.5% and ranged between 84 and 95% for individual questions. Conclusions None of the clinical prediction tools fulfilled the requirements to provide women with all the necessary information for informed decision-making. Information on individual prognosis was well understood and can be included in patient decision aids

Associations between older adults’ physical fitness level and their engagement in different types of physical activity: cross-sectional results from the OUTDOOR ACTIVE study

Objectives To describe the prevalence of different types of physical activities and to explore the association between engagement in these physical activities and performance in the physical fitness dimensions among older adults living in Bremen, Germany.Design Cross-sectional study.Setting Twelve subdistricts in Bremen, Germany.Participants 1583 non-institutionalised adults aged 65–75 years residing in one of the 12 subdistricts in Bremen, Germany (53.1% female).Primary outcome measures Level of physical fitness in five dimensions: handgrip strength (hand dynamometry), lower muscle strength (30 s-chair stand test), aerobic endurance (2 min-step test), lower body flexibility (sit-and-reach test), and upper body flexibility (back scratch test); classified using normative values.Results In this study population, home-based activities (housework, gardening) and activities for transport (walking, cycling) were performed by nearly all the participants, while leisure activities were less prevalent. Logistic regression revealed a positive association between being in or over norm in handgrip strength with cycling (OR: 1.56, 95% CI: 1.13 to 2.15), hiking/running (OR: 1.50, 95% CI: 1.05 to 2.16) and other sports (OR: 3.22, 95% CI: 1.37 to 7.56). Lower muscle strength was positively related to cycling (OR: 1.91, 95% CI: 1.37 to 2.65), gym training (OR: 1.62, 95% CI: 1.16 to 2.26) and dancing (OR: 2.15, 95% CI: 1.00 to 4.61). Aerobic endurance was positively associated with cycling (OR: 1.90, 95% CI: 1.37 to 2.65), gym training (OR: 1.68, 95% CI: 1.20 to 2.36), aerobics (OR: 1.64, 95% CI: 1.19 to 2.26), dancing (OR: 2.62, 95% CI: 1.10 to 6.22) and ball sports (OR: 2.07, 95% CI: 1.30 to 3.29). Apart from housework and upper body flexibility (OR: 0.39, 95% CI: 0.19 to 0.78), flexibility dimensions did not show any significant associations.Conclusions While muscle strength dimensions and aerobic endurance were associated with several physical activities, flexibility dimensions were associated with none of the investigated activities apart from housework. Especially cycling and leisure activities (eg, hiking/ running, gym training, aerobics, dancing) showed great potential for sustaining and increasing physical fitness in older age

Adherence to Lifestyle Recommendations for Bone Health in Older Adults with and without Osteoporosis: Cross-Sectional Results of the OUTDOOR ACTIVE Study

Sustaining good bone health into older age is key for preventing osteoporosis. Bone health is associated with several lifestyle factors. This study investigates older adults’ adherence to bone health-promoting lifestyle recommendations dependent on osteoporosis status. Cross-sectional data of 1610 community-dwelling older adults (65–75 years) residing in Bremen, Germany (53.4% female) were included. The Osteoporosis Self-Assessment Tool and self-reported osteoporosis diagnosis were used to classify participants by osteoporosis status (low risk, high risk, diagnosis). Adherence to bone health recommendations regarding calcium and vitamin D intake, sun exposure, alcohol consumption, resistance/weight-bearing exercise, and physical activity were assessed. Descriptive statistics were applied, stratified by sex and osteoporosis status. A total of 91 women (10.6%) and 15 men (2.0%) reported an osteoporosis diagnosis, 457 women (43.2%) and 311 men (41.4%) were classified as having a high risk, and 311 women (36.2%) and 425 men (56.6%) as having a low risk. Adherence to bone health recommendations was high for calcium intake (93.3–100.0%), vitamin D intake (77.8–93.3%), and sun exposure (86.7–97.7%). Lower adherence was observed regarding resistance/weight-bearing exercise (36.3–54.4%), physical activity (14.3–57.7%), and alcohol consumption (40.0–72.4%). In conclusion, tailored prevention strategies are needed that focus on older adults with an osteoporosis diagnosis or who are at high risk

Generation and validation of ActiGraph GT3X+ accelerometer cut-points for assessing physical activity intensity in older adults. The OUTDOOR ACTIVE validation study.

The study of physical activity in older adults is becoming more and more relevant. For evaluation of physical activity recommendations, intensity-specific accelerometer cut-points are utilized. However, research on accelerometer cut-points for older adults is still scarce. The aim of the study was to generate placement-specific cut-points of ActiGraph GT3X+ activity counts and raw measures of acceleration to determine physical activity intensity in older adults. A further aim was to compare the validity of the generated cut-points for a range of different physical activities. The study was a single experimental trial using a convenience sample. Study participants were 20 adults aged 59 to 73 years. Accelerometers were worn at six different placements (one on each wrist, one on each ankle, and two at the hip) and breath-by-breath indirect calorimetry was used as the reference for energy. The experiment comprised of two parts; a) The first required participants to walk on a treadmill at incremental speeds (3.0–5.0 km·h(-1)), and b) Five different everyday activities (reading, cleaning, shopping, cycling, aerobics) were staged in the laboratory setting. Accelerometer cut-points (activity counts, raw data) were derived for each of the investigated placements by linear regression using the treadmill part. Performance of the cut-points was assessed by applying the cut-points to the everyday activities. We provide cut-points for six placements and two accelerometer metrics in the specific age group. However, the derived cut-points did not outperform published ones. More research and innovative approaches are needed for improving internal and external validity of research results across populations and age groups

Your personalized protein structure: Andrei N. Lupas fused to GCN4 adaptors

AbstractThis work presents a protein structure that has been designed purely for aesthetic reasons, symbolizing decades of coiled-coil research and praising its most fundamental model system, the GCN4 leucine zipper. The GCN4 leucine zipper is a highly stable coiled coil which can be tuned to adopt different oligomeric states via mutation of its core residues. For these reasons it is used in structural studies as a stabilizing fusion adaptor. On the occasion of the 50th birthday of Andrei N. Lupas, we used it to create the first personalized protein structure: we fused the sequence ANDREI-N-LVPAS in heptad register to trimeric GCN4 adaptors and determined its structure by X-ray crystallography. The structure demonstrates the robustness and versatility of GCN4 as a fusion adaptor. We learn how proline can be accommodated in trimeric coiled coils, and put the structure into the context of the other GCN4-fusion structures known to date