Elucidating the mechano-molecular dynamics of TRAP activity using CRISPR/Cas9 mediated fluorescent reporter mice.

Osteoclasts are essential for bone remodeling by adapting their resorptive activity in response to their mechanical in vivo environment. However, the molecular mechanisms underlying this process remain unclear. Here, we demonstrated the role of tartrate-resistant acid phosphatase (TRAP, Acp5), a key enzyme secreted by osteoclasts, in bone remodeling and mechanosensitivity. Using CRISPR/Cas9 reporter mice, we demonstrated bone cell reporter (BCRIbsp/Acp5) mice feature fluorescent TRAP-deficient osteoclasts and examined their activity during mechanically driven trabecular bone remodeling. Although BCRIbsp/Acp5 mice exhibited trabecular bone impairments and reduced resorption capacity in vitro, RNA sequencing revealed unchanged levels of key osteoclast-associated genes such as Ctsk, Mmp9, and Calcr. These findings, in conjunction with serum carboxy-terminal collagen crosslinks (CTX) and in vivo mechanical loading outcomes collectively indicated an unaltered bone resorption capacity of osteoclasts in vivo. Furthermore, we demonstrated similar mechanoregulation during trabecular bone remodeling in BCRIbsp/Acp5 and wild-type (WT) mice. Hence, this study provides valuable insights into the dynamics of TRAP activity in the context of bone remodeling and mechanosensation.

Mechanisms of programmed cell death associated to severe dengue in human renal lesions.

Dengue virus (DENV) is a global health problem. Severe dengue can manifest with hemorrhage and signs of organ dysfunction, including the kidneys. The innate immune system is an important barrier against arbovirus infection and, specifically in dengue, the cytokines IL1ß and IL18 and caspase-1 activation make up a set of host immune strategies. Cell death mechanisms include pyroptosis, necroptosis and autophagy, each with peculiar markers: gasdermin, RIPK3/MLKL, LC3, respectively. In DENV infection, necrosis and apoptosis are involved and, when infecting monocytes and macrophages in vitro, DENV is capable of inducing pyroptosis. Our objective was to explore the presence of markers of necroptosis, pyroptosis and autophagy in renal lesions caused by DENV. MATERIAL AND METHODS: twenty specimens of lesions from patients who died due to DENV infection, from the pathology department of Hospital Guilherme Álvaro, Santos, SP, were subjected to histological and immunohistochemical studies. Histological sections were stained with hematoxylin-eosin to evaluate tissue changes or collected for research with antibodies: anti-DENV (Instituto Evandro Chagas-PA), RIPK3 (NBP2-45592), MLKL (ab184718), gasdermin D (#36425 ), LC3 (14600-AP), caspase 1 (#98033), IL1ß (AF201-NA) and IL18 (SC6178). Semi-quantitative analysis was performed on 20 glomeruli and evaluation on tubules and mononuclear cells. This study was approved by the ethics committee of the USP Faculty of Medicine. RESULTS: histological analysis demonstrated glomerular congestion, glomerulitis (medium to severe), acute kidney injury and hyalinization of the glomeruli. Viral antigens were visualized on mononuclear cells. LC3 (autophagy) expression ranged from moderate to intense (++/+++) in glomeruli, tubules and mononuclear cells. The expression of gasdermin (pyroptosis) was mild (+) in most cases in the glomeruli and moderate (++) in the tubules. RIPK3 and MLKL (necroptosis) mild in tubules and mononuclear cells (+). The expression of the cytokines IL1ß and IL18 and caspase 1 was moderate (++). Statistical analysis showed greater expression of LC3 over the others. CONCLUSIONS: Our results contribute to the understanding of the pathogenesis of renal involvement in severe dengue, considering the likely anti-viral mechanism of autophagy. To a lesser extent, pyroptosis is also present, corroborating previous data.

Phase 1/2 AAV5-hRKp.RPGR (Botaretigene Sparoparvovec) Gene Therapy: Safety and Efficacy in RPGR-Associated X-Linked Retinitis Pigmentosa.

PURPOSE: To assess the safety and efficacy of AAV5-hRKp.RPGR in participants with retinitis pigmentosa GTPase regulator (RPGR)-associated X-linked retinitis pigmentosa (XLRP). DESIGN: Open-label, phase 1/2 dose escalation/expansion study (ClinicalTrials.gov Identifier: NCT03252847). METHODS: Males (≥5 years old) with XLRP-RPGR were evaluated. In the dose escalation phase, subretinal AAV5-hRKp.RPGR (low: 1.0 × 1011 vg/ml; intermediate: 2.0 × 1011 vg/ml; high: 4.0 × 1011 vg/ml) was administered to the poorer-seeing eye (n = 10). Dose confirmation (intermediate dose) was carried out in 3 pediatric participants. In the dose expansion phase, 36 participants were randomized 1:1:1 to immediate (low or intermediate dose) or deferred (control) treatment. The primary outcome was safety. Secondary efficacy outcomes included static perimetry, microperimetry, vision-guided mobility, best corrected visual acuity, and contrast sensitivity. Safety and efficacy outcomes were assessed for 52 weeks for immediate treatment participants and 26 weeks for control participants. RESULTS: AAV5-hRKp.RPGR was safe and well tolerated, with no reported dose-limiting events. Most adverse events (AEs) were transient and related to the surgical procedure, resolving without intervention. Two serious AEs were reported with immediate treatment (retinal detachment, uveitis). A third serious AE (increased intraocular pressure) was reported outside the reporting period. All ocular inflammation-related AEs responded to corticosteroids. Treatment with AAV5-hRKp.RPGR resulted in improvements in retinal sensitivity and functional vision compared with the deferred group at Week 26; similar trends were observed at Week 52. CONCLUSIONS: AAV5-hRKp.RPGR demonstrated an anticipated and manageable AE profile through 52 weeks. Safety and efficacy findings support investigation in a phase 3 trial.

Our Ways, Your Ways, Both Ways - a multi-disciplinary collaboration to develop, embed and evaluate a model of social and emotional wellbeing care for Aboriginal and Torres Strait Islander young people who experience detention - Phase 1.

The National Strategic Framework for Aboriginal and Torres Strait Islander Peoples' Mental Health and Social and Emotional Wellbeing identifies building a strong Aboriginal and Torres Strait Islander led evidence-base to inform care as a key priority. Aboriginal and/or Torres Strait Islander adolescents in contact with the criminal justice system are a highly vulnerable group of Australians, with substantial unmet needs. There is limited evidence to inform culturally appropriate models of care that meet the social and emotional wellbeing needs of justice-involved Aboriginal and/or Torres Strait Islander adolescents. This project aims to develop, implement and evaluate an in-reach and community transitional model of social and emotional wellbeing care for Aboriginal and/or Torres Strait Islander adolescents (10-17 years old) who experience detention through close engagement with Aboriginal and/or Torres Strait Islander youth, Elders, researchers, practitioners and community members, and by drawing on culturally informed practice and knowledge systems. The project is based on a multi-level mixed methods design, with a strong focus on ongoing project evaluation (based on the Ngaa-bi-nya framework) and co-design. Co-design is facilitated through culturally safe and trauma informed participatory processes based on development of strong partnerships from project initiative, design, implementation and evaluation. Application of the landscape domain of the Ngaa-bi-nya framework for Aboriginal and Torres Strait Islander program evaluation will be explored in Phase one. Aboriginal and Torres Strait Islander adolescents with experience in detention will be engaged through one-on-one interviews with data collection through the Growth and Empowerment Measure (GEM) Youth (which will be adapted from the adult version and validated as part of this study), the Kessler Psychological Distress Scale (K-10), questions around alcohol and drug use, and narrative interviews exploring experience. Qualitative data will be analyzed using an inductive thematic approach, structured within the framework of the Ngaa-bi-nya landscape prompts. Quantitative data will be analyzed using descriptive statistics to provide a profile of the cohort. Findings from Phase one will be used to inform the development of a model of social and emotional wellbeing care that will be implemented and evaluated in Phase two.

A multi-stack registration technique to improve measurement accuracy and precision across longitudinal HR-pQCT scans.

BACKGROUND: Recent applications of high-resolution peripheral quantitative computed tomography (HR-pQCT) have demonstrated that changes in local bone remodelling can be quantified in vivo using longitudinal three-dimensional image registration. However, certain emerging applications, such as fracture healing and joint analysis, require larger multi-stack scan regions that can result in stack shift image artifacts. These artifacts can be detrimental to the accurate alignment of the bone structure across multiple timepoints. The purpose of this study was to establish a multi-stack registration protocol for evaluating longitudinal HR-pQCT images and to assess the accuracy and precision error in comparison with measures obtained using previously established three-dimensional longitudinal registration. METHODS: Three same day multi-stack HR-pQCT scans of the radius (2 stacks in length) and tibia (3 stacks in length) were obtained from 39 healthy individuals who participated in a previous reproducibility study. A fully automated multi-stack registration algorithm was developed to re-align stacks within a scan by leveraging slight offsets between longitudinal scans. Stack shift severity before and after registration was quantified using a newly proposed stack-shift severity score. The false discovery rate for bone remodelling events and precision error of bone morphology and micro-finite element analysis parameters were compared between longitudinally registered scans with and without the addition of multi-stack registration. RESULTS: Most scans (82 %) improved in stack alignment or maintained the lowest stack shift severity score when multi-stack registration was implemented. The false discovery rate of bone remodelling events significantly decreased after multi-stack registration, resulting in median false detection of bone formation and resorption fractions between 3.2 to 7.5 % at the radius and 3.4 to 5.3 % at the tibia. Further, precision error was significantly reduced or remained unchanged in all standard bone morphology and micro-finite element analysis parameters, except for total and trabecular cross-sectional areas. CONCLUSION: Multi-stack registration is an effective strategy for accurately aligning multi-stack HR-pQCT scans without modification of the image acquisition protocol. The algorithm presented here is a viable approach for performing accurate morphological analysis on multi-stack HR-pQCT scans, particularly for advanced application investigating local bone remodelling in vivo.

A Correspondence-Based Network Approach for Groupwise Analysis of Patient-Specific Spatiotemporal Data.

Methods for statistically analyzing patient-specific data that vary both spatially and over time are currently either limited to summary statistics or require elaborate surface registration. We propose a new method, called correspondence-based network analysis, which leverages particle-based shape modeling to establish correspondence across a population and preserve patient-specific measurements and predictions through statistical analysis. Herein, we evaluated this method using three published datasets of the hip describing cortical bone thickness of the proximal femur, cartilage contact stress, and dynamic joint space between control and patient cohorts to evaluate activity- and group-based differences, as applicable, using traditional statistical parametric mapping (SPM) and our proposed spatially considerate correspondence-based network analysis approach. The network approach was insensitive to correspondence density, while the traditional application of SPM showed decreasing area of the region of significance with increasing correspondence density. In comparison to SPM, the network approach identified broader and more connected regions of significance for all three datasets. The correspondence-based network analysis approach identified differences between groups and activities without loss of subject and spatial specificity which could improve clinical interpretation of results.

Precision of bone mechanoregulation assessment in humans using longitudinal high-resolution peripheral quantitative computed tomography in vivo.

Local mechanical stimuli in the bone microenvironment are essential for the homeostasis and adaptation of the skeleton, with evidence suggesting that disruption of the mechanically-driven bone remodelling process may lead to bone loss. Longitudinal clinical studies have shown the combined use of high-resolution peripheral quantitative computed tomography (HR-pQCT) and micro-finite element analysis can be used to measure load-driven bone remodelling in vivo; however, quantitative markers of bone mechanoregulation and the precision of these analyses methods have not been validated in human subjects. Therefore, this study utilised participants from two cohorts. A same-day cohort (n = 33) was used to develop a filtering strategy to minimise false detections of bone remodelling sites caused by noise and motion artefacts present in HR-pQCT scans. A longitudinal cohort (n = 19) was used to develop bone imaging markers of trabecular bone mechanoregulation and characterise the precision for detecting longitudinal changes in subjects. Specifically, we described local load-driven formation and resorption sites independently using patient-specific odds ratios (OR) and 99 % confidence intervals. Conditional probability curves were computed to link the mechanical environment to the remodelling events detected on the bone surface. To quantify overall mechanoregulation, we calculated a correct classification rate measuring the fraction of remodelling events correctly identified by the mechanical signal. Precision was calculated as root-mean-squared averages of the coefficient of variation (RMS-SD) of repeated measurements using scan-rescan pairs at baseline combined with a one-year follow-up scan. We found no significant mean difference (p < 0.01) between scan-rescan conditional probabilities. RMS-SD was 10.5 % for resorption odds, 6.3 % for formation odds, and 1.3 % for correct classification rates. Bone was most likely to be formed in high-strain and resorbed in low-strain regions for all participants, indicating a consistent, regulated response to mechanical stimuli. For each percent increase in strain, the likelihood of bone resorption decreased by 2.0 ± 0.2 %, and the likelihood of bone formation increased by 1.9 ± 0.2 %, totalling 38.3 ± 1.1 % of strain-driven remodelling events across the entire trabecular compartment. This work provides novel robust bone mechanoregulation markers and their precision for designing future clinical studies.

Guidance on the use of complex systems models for economic evaluations of public health interventions.

To help health economic modelers respond to demands for greater use of complex systems models in public health. To propose identifiable features of such models and support researchers to plan public health modeling projects using these models. A working group of experts in complex systems modeling and economic evaluation was brought together to develop and jointly write guidance for the use of complex systems models for health economic analysis. The content of workshops was informed by a scoping review. A public health complex systems model for economic evaluation is defined as a quantitative, dynamic, non-linear model that incorporates feedback and interactions among model elements, in order to capture emergent outcomes and estimate health, economic and potentially other consequences to inform public policies. The guidance covers: when complex systems modeling is needed; principles for designing a complex systems model; and how to choose an appropriate modeling technique. This paper provides a definition to identify and characterize complex systems models for economic evaluations and proposes guidance on key aspects of the process for health economics analysis. This document will support the development of complex systems models, with impact on public health systems policy and decision making.

"If you were a cow, what would you want?" Findings from participatory workshops with dairy farmers.

When looking to improve animal welfare, it is essential to understand the viewpoints of stakeholders in the industry. Previous research has engaged stakeholders such as farmers, veterinarians, and the public to better understand future dairy industry directions. However, the use of perspective-taking as a tool to overcome farmer's focus on current barriers to the industry has not been explored. Therefore, the aims of this study were to have farmers take the perspective of the cow, to elicit: (1) farm elements that are essential to create a cow-centric dairy system, and (2) how this system can be achieved in the next 50 years. To do this, we employed participatory methods (photo elicitation, timelining) to generate discussion with 12 New Zealand dairy farmers. Pasture access is frequently cited as an integral component for promoting good welfare in dairy cows, thus, this cohort of farmers was considered a good case study as they all had experience managing cows in grazing systems. Participants were asked to create a cow-centric farm design, and a pathway to implementation. Following thematic analysis of the multiple data sources, the results are presented under the following three themes: (1) cow-centric farm designs (environmental considerations, cow handling and care), (2) timeline to achieve and sequence of implementation ("low hanging fruit" and long-term investments), and (3) assumptions and resources needed (including financial considerations, technology, farmer buy-in, regulations and enforcement and other stakeholder involvement). By better understanding how current industry practice aligns, and also differs, from what the cow would want, we can work towards future management systems that incorporate the requirements of all stakeholders, including the cow.

Statistical multi-level shape models for scalable modeling of multi-organ anatomies.

Statistical shape modeling is an indispensable tool in the quantitative analysis of anatomies. Particle-based shape modeling (PSM) is a state-of-the-art approach that enables the learning of population-level shape representation from medical imaging data (e.g., CT, MRI) and the associated 3D models of anatomy generated from them. PSM optimizes the placement of a dense set of landmarks (i.e., correspondence points) on a given shape cohort. PSM supports multi-organ modeling as a particular case of the conventional single-organ framework via a global statistical model, where multi-structure anatomy is considered as a single structure. However, global multi-organ models are not scalable for many organs, induce anatomical inconsistencies, and result in entangled shape statistics where modes of shape variation reflect both within- and between-organ variations. Hence, there is a need for an efficient modeling approach that can capture the inter-organ relations (i.e., pose variations) of the complex anatomy while simultaneously optimizing the morphological changes of each organ and capturing the population-level statistics. This paper leverages the PSM approach and proposes a new approach for correspondence-point optimization of multiple organs that overcomes these limitations. The central idea of multilevel component analysis, is that the shape statistics consists of two mutually orthogonal subspaces: the within-organ subspace and the between-organ subspace. We formulate the correspondence optimization objective using this generative model. We evaluate the proposed method using synthetic shape data and clinical data for articulated joint structures of the spine, foot and ankle, and hip joint.

Acetabular labrum and cartilage contact mechanics during pivoting and walking tasks in individuals with cam femoroacetabular impingement syndrome.

Femoroacetabular impingement syndrome (FAIS) is a motion-related pathology of the hip characterized by pain, morphological abnormalities of the proximal femur, and an elevated risk of joint deterioration and hip osteoarthritis. Activities that require deep flexion are understood to induce impingement in cam FAIS patients, however, less demanding activities such as walking and pivoting may induce pain as well as alterations in kinematics and joint stability. Still, the paucity of quantitative descriptions of cam FAIS has hindered understanding underlying hip joint mechanics during such activities. Previous in silico studies have employed generalized model geometry or kinematics to simulate impingement between the femur and acetabulum, which may not accurately capture the interplay between morphology and motion. In this study, we utilized models with participant-specific bone and articular soft tissue anatomy and kinematics measured by dual-fluoroscopy to compare hip contact mechanics of cam FAIS patients to controls during four activities of daily living (internal/external pivoting and level/incline walking). Averaged across the gait cycle during incline walking, patients displayed increased strain in the anterior joint (labrum strain: p-value = 0.038, patients: 11.7 ± 6.7 %, controls: 5.0 ± 3.6 %; cartilage strain: p-value = 0.029, patients: 9.1 ± 3.3 %, controls: 4.2 ± 2.3). Patients also exhibited increased average anterior cartilage strains during external pivoting (p-value = 0.039; patients: 13.0 ± 9.2 %, controls: 3.9 ± 3.2 %]). No significant differences between patient and control contact area and strain were found for level walking and internal pivoting. Our study provides new insights into the biomechanics of cam FAIS, including spatiotemporal hip joint contact mechanics during activities of daily living.

Patients with cam-type femoroacetabular impingement demonstrate increased change in bone-to-bone distance during walking: A dual fluoroscopy study.

Cam-type femoroacetabular impingement (FAI) syndrome is a painful, structural hip disorder. Herein, we investigated hip joint mechanics through in vivo, dynamic measurement of the bone-to-bone distance between the femoral head and acetabulum in patients with cam FAI syndrome and morphologically screened controls. We hypothesized that individuals with cam FAI syndrome would have larger changes in bone-to-bone distance compared to the control group, which we would interpret as altered joint mechanics as signified by greater movement of the femoral head as it articulates within the acetabulum. Seven patients with cam FAI syndrome and 11 asymptomatic individuals with typical morphology underwent dual fluoroscopy imaging during level and inclined walking (upward slope). The change in bone-to-bone distance between femoral and acetabular bone surfaces was evaluated for five anatomical regions of the acetabulum at each timepoint of gait. Linear regression analysis of the bone-to-bone distance considered two within-subject factors (activity and region) and one between-subjects factor (group). Across activities, the change in minimum bone-to-bone distance was 1.38-2.54 mm for the cam FAI group and 1.16-1.84 mm for controls. In all regions except the anterior-superior region, the change in bone-to-bone distance was larger in the cam group than the control group (p ≤ 0.024). An effect of activity was detected only in the posterior-superior region where larger changes were noted during level walking than incline walking. Statement of clinical significance: Patients with cam FAI syndrome exhibit altered hip joint mechanics during the low-demand activity of walking; these alterations could affect load transmission, and contribute to pain, tissue damage, and osteoarthritis.

Motion grading of high-resolution quantitative computed tomography supported by deep convolutional neural networks.

Image quality degradation due to subject motion confounds the precision and reproducibility of measurements of bone density, morphology and mechanical properties from high-resolution peripheral quantitative computed tomography (HR-pQCT). Time-consuming operator-based scoring of motion artefacts remains the gold standard to determine the degree of acceptable motion. However, due to the subjectiveness of manual grading, HR-pQCT scans of poor quality, which cannot be used for analysis, may be accepted upon initial review, leaving patients with incomplete or inaccurate imaging results. Convolutional Neural Networks (CNNs) enable fast image analysis with relatively few pre-processing requirements in an operator-independent and fully automated way for image classification tasks. This study aimed to develop a CNN that can predict motion scores from HR-pQCT images, while also being aware of uncertain predictions that require manual confirmation. The CNN calculated motion scores within seconds and achieved a high F1-score (86.8 ± 2.8 %), with good precision (87.5 ± 2.7 %), recall (86.7 ± 2.9 %) and a substantial agreement with the ground truth measured by Cohen's kappa (κ = 68.6 ± 6.2 %); motion scores of the test dataset were predicted by the algorithm with comparable accuracy, precision, sensitivity and agreement as by the operators (p > 0.05). This post-processing approach may be used to assess the effect of motion scores on microstructural analysis and can be immediately implemented into clinical protocols, significantly reducing the time for quality assessment and control of HR-pQCT scans.

Clinical observation of diminished bone quality and quantity through longitudinal HR-pQCT-derived remodeling and mechanoregulation.

High resolution peripheral quantitative computed tomography (HR-pQCT) provides methods for quantifying volumetric bone mineral density and microarchitecture necessary for early diagnosis of bone disease. When combined with a longitudinal imaging protocol and finite element analysis, HR-pQCT can be used to assess bone formation and resorption (i.e., remodeling) and the relationship between this remodeling and mechanical loading (i.e., mechanoregulation) at the tissue level. Herein, 25 patients with a contralateral distal radius fracture were imaged with HR-pQCT at baseline and 9-12 months follow-up: 16 patients were prescribed vitamin D3 with/without calcium supplement based on a blood biomarker measures of bone metabolism and dual-energy X-ray absorptiometry image-based measures of normative bone quantity which indicated diminishing (n = 9) or poor (n = 7) bone quantity and 9 were not. To evaluate the sensitivity of this imaging protocol to microstructural changes, HR-pQCT images were registered for quantification of bone remodeling and image-based micro-finite element analysis was then used to predict local bone strains and derive rules for mechanoregulation. Remodeling volume fractions were predicted by both average values of trabecular and cortical thickness and bone mineral density (R2 > 0.8), whereas mechanoregulation was affected by dominance of the arm and group classification (p < 0.05). Overall, longitudinal, extended HR-pQCT analysis enabled the identification of changes in bone quantity and quality too subtle for traditional measures.

BMI trajectories, morbidity, and mortality in England: a two-step approach to estimating consequences of changes in BMI.

OBJECTIVE: BMI is known to have an association with morbidities and mortality. Many studies have argued that identifying health risks using single BMI measures has limitations, particularly in older adults, and that changes in BMI can help to identify risks. This study identifies distinct BMI trajectories and their association with the risks of a range of morbidities and mortality. METHODS: The English Longitudinal Study of Aging provides data on BMI, mortality, and morbidities between 1998 and 2015, sampled from adults over 50 years of age. This study uses a growth-mixture model and discrete-time survival analysis, combined using a two-step approach, which is novel in this setting, to the authors' knowledge. RESULTS: This study identified four trajectories: "stable overweight," "elevated BMI," "increasing BMI," and "decreasing BMI." No differences in mortality, cancer, or stroke risk were found between these trajectories. BMI trajectories were significantly associated with the risks of diabetes, asthma, arthritis, and heart problems. CONCLUSIONS: These results emphasize the importance of looking at change in BMI alongside most recent BMI; BMI trajectories should be considered where possible when assessing health risks. The results suggest that established BMI thresholds should not be used in isolation to identify health risks, particularly in older adults.

Development of a Web-Based, Guided Self-help, Acceptance and Commitment Therapy-Based Intervention for Weight Loss Maintenance: Evidence-, Theory-, and Person-Based Approach.

BACKGROUND: The long-term impact and cost-effectiveness of weight management programs depend on posttreatment weight maintenance. There is growing evidence that interventions based on third-wave cognitive behavioral therapy, particularly acceptance and commitment therapy (ACT), could improve long-term weight management; however, these interventions are typically delivered face-to-face by psychologists, which limits the scalability of these types of intervention. OBJECTIVE: The aim of this study is to use an evidence-, theory-, and person-based approach to develop an ACT-based intervention for weight loss maintenance that uses digital technology and nonspecialist guidance to minimize the resources needed for delivery at scale. METHODS: Intervention development was guided by the Medical Research Council framework for the development of complex interventions in health care, Intervention Mapping Protocol, and a person-based approach for enhancing the acceptability and feasibility of interventions. Work was conducted in two phases: phase 1 consisted of collating and analyzing existing and new primary evidence and phase 2 consisted of theoretical modeling and intervention development. Phase 1 included a synthesis of existing evidence on weight loss maintenance from previous research, a systematic review and network meta-analysis of third-wave cognitive behavioral therapy interventions for weight management, a qualitative interview study of experiences of weight loss maintenance, and the modeling of a justifiable cost for a weight loss maintenance program. Phase 2 included the iterative development of guiding principles, a logic model, and the intervention design and content. Target user and stakeholder panels were established to inform each phase of development, and user testing of successive iterations of the prototype intervention was conducted. RESULTS: This process resulted in a guided self-help ACT-based intervention called SWiM (Supporting Weight Management). SWiM is a 4-month program consisting of weekly web-based sessions for 13 consecutive weeks followed by a 4-week break for participants to reflect and practice their new skills and a final session at week 18. Each session consists of psychoeducational content, reflective exercises, and behavioral experiments. SWiM includes specific sessions on key determinants of weight loss maintenance, including developing skills to manage high-risk situations for lapses, creating new helpful habits, breaking old unhelpful habits, and learning to manage interpersonal relationships and their impact on weight management. A trained, nonspecialist coach provides guidance for the participants through the program with 4 scheduled 30-minute telephone calls and 3 further optional calls. CONCLUSIONS: This comprehensive approach facilitated the development of an intervention that is based on scientific theory and evidence for supporting people with weight loss maintenance and is grounded in the experiences of the target users and the context in which it is intended to be delivered. The intervention will be refined based on the findings of a planned pilot randomized controlled trial.

Prediction of femoral head coverage from articulated statistical shape models of patients with developmental dysplasia of the hip.

Developmental dysplasia of the hip (DDH) is commonly described as reduced femoral head coverage due to anterolateral acetabular deficiency. Although reduced coverage is the defining trait of DDH, more subtle and localized anatomic features of the joint are also thought to contribute to symptom development and degeneration. These features are challenging to identify using conventional approaches. Herein, we assessed the morphology of the full femur and hemi-pelvis using an articulated statistical shape model (SSM). The model determined the morphological and pose-based variations associated with DDH in a population of Japanese females and established which of these variations predict coverage. Computed tomography (CT) images of 83 hips from 47 patients were segmented for input into a correspondence-based SSM. The dominant modes of variation in the model initially represented scale and pose. After removal of these factors through individual bone alignment, femoral version and neck-shaft angle, pelvic curvature, and acetabular version dominated the observed variation. Femoral head oblateness and prominence of the acetabular rim and various muscle attachment sites of the femur and hemi-pelvis were found to predict 3D CT-based coverage measurements (R2 = 0.5-0.7 for the full bones, R2 = 0.9 for the joint). Statement of Clinical Significance: Currently, clinical measurements of DDH only consider the morphology of the acetabulum. However, the results of this study demonstrated that variability in femoral head shape and several muscle attachment sites were predictive of femoral head coverage. These morphological differences may provide insight into improved clinical diagnosis and surgical planning based on functional adaptations of patients with DDH.

Land Management Legacy Affects Abundance and Function of the acdS Gene in Wheat Root Associated Pseudomonads.

Land management practices can vastly influence belowground plant traits due to chemical, physical, and biological alteration of soil properties. Beneficial Pseudomonas spp. are agriculturally relevant bacteria with a plethora of plant growth promoting (PGP) qualities, including the potential to alter plant physiology by modulating plant produced ethylene via the action of the bacterial enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase (acdS). This study evaluated the impact of land management legacy on the selection and function of wheat root associated culturable pseudomonad isolates. Three distinct previous land uses prior to wheat culture (grassland, arable, and bare fallow) were tested and culturable pseudomonad abundance, phylogeny (gyrB and acdS genes), function (ACC deaminase activity), and the co-selection of acdS with other PGP genes examined. The pseudomonad community could to some extent be discriminated based on previous land use. The isolates from rhizosphere and root compartments of wheat had a higher acdS gene frequency than the bulk soil, particularly in plants grown in soil from the bare fallow treatment which is known to have degraded soil properties such as low nutrient availability. Additionally, other genes of interest to agriculture encoding anti-fungal metabolites, siderophores, and genes involved in nitrogen metabolism were highly positively associated with the presence of the acdS gene in the long-term arable treatment in the genomes of these isolates. In contrast, genes involved in antibiotic resistance and type VI secretion systems along with nitrogen cycling genes were highly positively correlated with the acdS gene in bare fallow isolated pseudomonad. This highlights that the three land managements prior to wheat culture present different selection pressures that can shape culturable pseudomonad community structure and function either directly or indirectly via the influence of wheat roots.

Metagenomic approaches reveal differences in genetic diversity and relative abundance of nitrifying bacteria and archaea in contrasting soils.

The abundance and phylogenetic diversity of functional genes involved in nitrification were assessed in Rothamsted field plots under contrasting management regimes-permanent bare fallow, grassland, and arable (wheat) cultivation maintained for more than 50 years. Metagenome and metatranscriptome analysis indicated nitrite oxidizing bacteria (NOB) were more abundant than ammonia oxidizing archaea (AOA) and bacteria (AOB) in all soils. The most abundant AOA and AOB in the metagenomes were, respectively, Nitrososphaera and Ca. Nitrososcosmicus (family Nitrososphaeraceae) and Nitrosospira and Nitrosomonas (family Nitrosomonadaceae). The most abundant NOB were Nitrospira including the comammox species Nitrospira inopinata, Ca. N. nitrificans and Ca. N. nitrosa. Anammox bacteria were also detected. Nitrospira and the AOA Nitrososphaeraceae showed most transcriptional activity in arable soil. Similar numbers of sequences were assigned to the amoA genes of AOA and AOB, highest in the arable soil metagenome and metatranscriptome; AOB amoA reads included those from comammox Nitrospira clades A and B, in addition to Nitrosomonadaceae. Nitrification potential assessed in soil from the experimental sites (microcosms amended or not with DCD at concentrations inhibitory to AOB but not AOA), was highest in arable samples and lower in all assays containing DCD, indicating AOB were responsible for oxidizing ammonium fertilizer added to these soils.

In Vivo Quantification of Hip Arthrokinematics during Dynamic Weight-bearing Activities using Dual Fluoroscopy.

Several hip pathologies have been attributed to abnormal morphology with an underlying assumption of aberrant biomechanics. However, structure-function relationships at the joint level remain challenging to quantify due to difficulties in accurately measuring dynamic joint motion. The soft tissue artifact errors inherent in optical skin marker motion capture are exacerbated by the depth of the hip joint within the body and the large mass of soft tissue surrounding the joint. Thus, the complex relationship between bone shape and hip joint kinematics is more difficult to study accurately than in other joints. Herein, a protocol incorporating computed tomography (CT) arthrography, three-dimensional (3D) reconstruction of volumetric images, dual fluoroscopy, and optical motion capture to accurately measure the dynamic motion of the hip joint is presented. The technical and clinical studies that have applied dual fluoroscopy to study form-function relationships of the hip using this protocol are summarized, and the specific steps and future considerations for data acquisition, processing, and analysis are described.