Feeder-free culture of human pluripotent stem cells drives MDM4-mediated gain of chromosome 1q.

Culture-acquired variants in human pluripotent stem cells (hPSCs) hinder their applications in research and clinic. However, the mechanisms that underpin selection of variants remain unclear. Here, through analysis of comprehensive karyotyping datasets from over 23,000 hPSC cultures of more than 1,500 lines, we explored how culture conditions shape variant selection. Strikingly, we identified an association of chromosome 1q gains with feeder-free cultures and noted a rise in its prevalence in recent years, coinciding with increased usage of feeder-free regimens. Competition experiments of multiple isogenic lines with and without a chromosome 1q gain confirmed that 1q variants have an advantage in feeder-free (E8/vitronectin), but not feeder-based, culture. Mechanistically, we show that overexpression of MDM4, located on chromosome 1q, drives variants' advantage in E8/vitronectin by alleviating genome damage-induced apoptosis, which is lower in feeder-based conditions. Our study explains condition-dependent patterns of hPSC aberrations and offers insights into the mechanisms of variant selection.

Bone-specific physical activity questionnaire-derived skeletal loading score predicts bone geometry, density, and strength indices: a cross-sectional study.

INTRODUCTION: The bone-specific physical activity questionnaire (BPAQ) provides a bone-relevant index of physical activity participation according to the mechanical loads experienced across the life span. MATERIALS AND METHODS: We aimed to examine relationships between historical bone-relevant physical activity and pQCT-derived parameters of bone strength. We recruited 532 healthy volunteers (277 males, 255 females) across a broad age range (4-97 years). Peripheral quantitative computed tomography (XCT-3000, Stratec, Germany) was used to examine volumetric bone density, area, and strength indices of the non-dominant tibia and radius. Exercise loading history from birth was determined using the past BPAQ (pBPAQ) score. Pearson correlation analysis was used to examine relationships between pBPAQ scores and pQCT parameters. RESULTS: Independent of sex, pBPAQ scores were associated with total density at the 38% and 66% tibial sites and the 66% radial site (r = 0.145-0.261, p ˂ 0.05), total area at the 38% tibial site and 4% and 66% radial sites (r = 0.129-0.156, p ˂ 0.05), and strength indices at all measured sites (r = 0.123-0.234, p < 0.05). CONCLUSION: We conclude that, independent of sex, historical bone-relevant physical activity is associated with pQCT-derived indices of bone strength, indicating that pBPAQ captures the characteristics of bone loading history that are likely to be relevant adaptive stimuli. A larger sample is required to examine the influence of age on this relationship.

c-Src activation as a potential marker of chemical-induced skin irritation using tissue-engineered skin equivalents.

Skin irritancy to topically applied chemicals is a significant problem that affects millions of people worldwide. New or modified chemical entities must be tested for potential skin irritancy by industry as part of the safety and toxicity profiling process. Many of these tests have now moved to a non-animal-based format to reduce experiments on animals. However, these tests for irritancy potential often rely on monolayer cultures of keratinocytes that are not representative of the skin architecture or tissue-engineered human skin equivalents (HSE) using complex multi-gene expression panels that are often cumbersome and not amenable for high throughput. Here, we show that human skin equivalents increase abundance of several phosphorylated kinases (c-Src, c-Jun, p53, GSK3α/ß) in response to irritant chemical stimulation by phosphokinase array analysis. Specific phosphorylation of c-SrcY419 was confirmed by immunoblotting and was plasma membrane-associated in basal/spinous cells by phospho-specific immunohistochemistry. Moreover, c-SrcY419 phosphorylation in response to the irritants lactic acid and capsaicin was inhibited by the c-Src inhibitors KB-SRC and betaine trimethylglycine. These data provide the first evidence for c-Src specific activation in response to chemical irritants and point to the development of new modes of rapid testing by immunodetection for first-pass screening of potential irritants.

Geometry and bone mineral density determinants of femoral neck strength changes following exercise.

Physical exercise induces spatially heterogeneous adaptation in bone. However, it remains unclear where the changes in BMD and geometry have the greatest impact on femoral neck strength. The aim of this study was to determine the principal BMD-and-geometry changes induced by exercise that have the greatest effect on femoral neck strength. Pre- and post-exercise 3D-DXA images of the proximal femur were collected of male participants from the LIFTMOR-M exercise intervention trial. Meshes with element-by-element correspondence were generated by morphing a template mesh to each bone to calculate changes in BMD and geometry. Finite element (FE) models predicted femoral neck strength changes under single-leg stance and sideways fall load. Partial least squares regression (PLSR) models were developed with BMD-only, geometry-only, and BMD-and-geometry changes to determine the principal modes that explained the greatest variation in neck strength changes. The PLSR models explained over 90% of the strength variation with 3 PLS components using BMD-only (R2 > 0.92, RMSE < 0.06 N) and 8 PLS components with geometry-only (R2 > 0.93, RMSE < 0.06 N). Changes in the superior neck and distal cortex were most important during single-leg stance while the superior neck, medial head, and lateral trochanter were most important during a sideways fall. Local changes in femoral neck and head geometry could differentiate the exercise groups from the control group. Exercise interventions may target BMD changes in the superior neck, inferior neck, and greater trochanter for improved femoral neck strength in single-leg stance and sideways fall.

The SUMO protease SENP3 regulates mitochondrial autophagy mediated by Fis1.

Mitochondria are unavoidably subject to organellar stress resulting from exposure to a range of reactive molecular species. Consequently, cells operate a poorly understood quality control programme of mitophagy to facilitate elimination of dysfunctional mitochondria. Here, we used a model stressor, deferiprone (DFP), to investigate the molecular basis for stress-induced mitophagy. We show that mitochondrial fission 1 protein (Fis1) is required for DFP-induced mitophagy and that Fis1 is SUMOylated at K149, an amino acid residue critical for Fis1 mitochondrial localization. We find that DFP treatment leads to the stabilization of the SUMO protease SENP3, which is mediated by downregulation of the E3 ubiquitin (Ub) ligase CHIP. SENP3 is responsible for Fis1 deSUMOylation and depletion of SENP3 abolishes DFP-induced mitophagy. Furthermore, preventing Fis1 SUMOylation by conservative K149R mutation enhances Fis1 mitochondrial localization. Critically, expressing a Fis1 K149R mutant restores DFP-induced mitophagy in SENP3-depleted cells. Thus, we propose a model in which SENP3-mediated deSUMOylation facilitates Fis1 mitochondrial localization to underpin stress-induced mitophagy.

The effect of low-intensity whole-body vibration with or without high-intensity resistance and impact training on risk factors for proximal femur fragility fracture in postmenopausal women with low bone mass: study protocol for the VIBMOR randomized controlled trial.

BACKGROUND: The prevailing medical opinion is that medication is the primary (some might argue, only) effective intervention for osteoporosis. It is nevertheless recognized that osteoporosis medications are not universally effective, tolerated, or acceptable to patients. Mechanical loading, such as vibration and exercise, can also be osteogenic but the degree, relative efficacy, and combined effect is unknown. The purpose of the VIBMOR trial is to determine the efficacy of low-intensity whole-body vibration (LIV), bone-targeted, high-intensity resistance and impact training (HiRIT), or the combination of LIV and HiRIT on risk factors for hip fracture in postmenopausal women with osteopenia and osteoporosis. METHODS: Postmenopausal women with low areal bone mineral density (aBMD) at the proximal femur and/or lumbar spine, with or without a history of fragility fracture, and either on or off osteoporosis medications will be recruited. Eligible participants will be randomly allocated to one of four trial arms for 9 months: LIV, HiRIT, LIV + HiRIT, or control (low-intensity, home-based exercise). Allocation will be block-randomized, stratified by use of osteoporosis medications. Testing will be performed at three time points: baseline (T0), post-intervention (T1; 9 months), and 1 year thereafter (T2; 21 months) to examine detraining effects. The primary outcome measure will be total hip aBMD determined by dual-energy X-ray absorptiometry (DXA). Secondary outcomes will include aBMD at other regions, anthropometrics, and other indices of bone strength, body composition, physical function, kyphosis, muscle strength and power, balance, falls, and intervention compliance. Exploratory outcomes include bone turnover markers, pelvic floor health, quality of life, physical activity enjoyment, adverse events, and fracture. An economic evaluation will also be conducted. DISCUSSION: No previous studies have compared the effect of LIV alone or in combination with bone-targeted HiRIT (with or without osteoporosis medications) on risk factors for hip fracture in postmenopausal women with low bone mass. Should either, both, or combined mechanical interventions be safe and efficacious, alternative therapeutic avenues will be available to individuals at elevated risk of fragility fracture who are unresponsive to or unwilling or unable to take osteoporosis medications. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry (www. anzctr.org.au ) (Trial number ANZCTR12615000848505, https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id = 368962 ); date of registration 14/08/2015 (prospectively registered). Universal Trial Number: U1111-1172-3652.

Determination of Chemical Irritation Potential Using a Defined Gene Signature Set on Tissue-Engineered Human Skin Equivalents.

There are no physical or visual manifestations that define skin sensitivity or irritation; a subjective diagnosis is made on the basis of the evaluation of clinical presentations, including burning, prickling, erythema, and itching. Adverse skin reaction in response to topically applied products is common and can limit the use of dermatological or cosmetic products. The purpose of this study was to evaluate the use of human skin equivalents based on immortalized skin keratinocytes and evaluate the potential of a 22-gene panel in combination with multivariate analysis to discriminate between chemicals known to act as irritants and those that do not. Test compounds were applied topically to full-thickness human skin equivalent or human ex vivo skin and gene signatures determined for known irritants and nonirritants. Principle component analysis showed the discriminatory potential of the 22-gene panel. Linear discrimination analysis, performed to further refine the gene set for a more high-throughput analysis, identified a putative seven-gene panel (IL-6, PTGS2, ATF3, TRPV3, MAP3K8, HMGB2, and matrix metalloproteinase gene MMP-3) that could distinguish potential irritants from nonirritants. These data offer promise as an in vitro prediction tool, although analysis of a large chemical test set is required to further evaluate the system.

A Nurse Educator's Perspective About Institutional Racism and White Supremacy in Nursing Education.

In the United States, nursing education programs at mostly white institutions are led by faculty that are 80% white. This absence of diversity is a symptom of systemic racism and white supremacy, reinforced through built systems of inequity and economic constraints that influence accessibility of nursing education programs. White cultural norms drive standards of professionalism and assimilation within nursing education programs. These standards are formulated from white cultural supremacy and contribute to the unconscious biases of nursing faculty. It is necessary to examine these biases to reduce potential and realized inequities for students of color in current nursing education programs. Challenging and changing these cultural norms can contribute to the dismantling of systemic racism and white supremacy in nursing education and the profession of nursing, thereby increasing the diversity of the professional workforce.

Assessment of femoral neck strength and bone mineral density changes following exercise using 3D-DXA images.

Physical exercise induces spatially heterogeneous bone changes in the proximal femur. Recent advances have enabled 3D dual-energy X-ray Absorptiometry (DXA)-based finite element (FE) models to estimate bone strength. However, its ability to detect exercise-induced BMD and strength changes is unclear. The aim of this study was to quantify the repeatability of vBMD and femoral neck strength obtained from 3D-DXA images and determine the changes due an exercise intervention. The DXA scans included pairs of same-day repeated scans from ten healthy females and pre- and post-exercise intervention scans of 26 males. FE models with element-by-element correspondence were generated by morphing a template mesh to each bone. BMD and femoral strength under single-leg-stance and sideways fall loading configurations were obtained for both groups and compared. In the repeated images, the total hip vBMD difference was 0.5 ± 2.5%. Element-by-element BMD differences reached 30 ± 50%. The strength difference in single-leg stance was 2.8 ± 13% and in sideways fall was 4.5% ± 19%. In the exercise group, strength changes were 6 ± 19% under single-leg stance and 1 ± 18% under sideways fall. vBMD parameters were weakly correlated to strength (R2 < 0.31). The exercise group had a mean bone accrual exceeding repeatability values in the femoral head and cortical regions. The case with the highest vBMD change (6.4%) caused 18% and -7% strength changes under single-leg stance and sideways fall. 3D-DXA technology can assess the effect of exercise interventions in large cohorts but its validity in individual cases should be interpreted with caution.

Factors that predict glycaemic response to sodium-glucose linked transporter (SGLT) inhibitors.

AIM: To determine the clinical and biochemical variables associated with change in HbA1c in patients with type 2 diabetes who start sodium-glucose linked transporter (SGLT) inhibitor therapy. METHODS: We performed a prospective cohort study (ACTRN12616000833460) of 48 adults (30 male, 18 female) with type 2 diabetes who attended a tertiary hospital diabetes clinic. Fasting serum and urine samples, collected during clinic visits prior to and at 1, 12 and 24 weeks after commencing SGLT inhibitor treatment, were analysed for HbA1c, electrolytes, urea, creatinine and glucose. RESULTS: After 12 weeks, SGLT inhibitor therapy was associated with respective median (97% CI) decreases in weight, blood pressure, HbA1c and urine albumin/creatinine ratio of 3.0 (1.7-3.4) kg, 8 (2-16)/4 (3-9) mmHg, 6 (3-14) mmol/mol and 0.69 (0.18-1.8) mg/mmol. These effects persisted to 24 weeks. Urinary frequency and genitourinary infection were common adverse effects. Baseline HbA1c and eGFR independently predicted ΔHbA1c at 12 weeks whereas only baseline HbA1c independently predicted ΔHbA1c at 24 weeks. Urinary fractional glucose excretion and change in fasting glucose 1 week after starting SGLT inhibitor did not contribute to prediction of glycaemic response. CONCLUSIONS: SGLT inhibitor therapy in a hospital clinic setting was associated with clinical improvements comparable to those observed in clinical trials but with higher incidence of genitourinary side-effects. Baseline HbA1c and eGFR, but not urine fractional glucose excretion, predicted glycaemic response.

Effects of supervised high-intensity resistance and impact training or machine-based isometric training on regional bone geometry and strength in middle-aged and older men with low bone mass: The LIFTMOR-M semi-randomised controlled trial.

INTRODUCTION: Few data exist on the effects of bone-targeted exercise on geometric and biomechanical indices of bone strength in men. The Lifting Intervention For Training Muscle and Osteoporosis Rehabilitation for Men (LIFTMOR-M) trial was designed to compare the efficacy and safety of two novel, supervised, twice-weekly, high-intensity exercise programs in middle-aged and older men with osteopenia and osteoporosis on musculoskeletal health and risk factors related to falls and fractures. The current report includes secondary outcomes of the LIFTMOR-M exercise intervention trial. PURPOSE: Our goal was to determine the effects of two supervised, twice-weekly, high-intensity exercise programs on bone geometry and strength of the proximal femur, and distal and proximal sites of the tibia and radius in middle-aged and older men with osteopenia and osteoporosis. METHODS: Generally-healthy men (≥45 years), with low lumbar spine (LS) and/or proximal femur areal bone mineral density (aBMD), were recruited from the community. Eligible participants were randomised to either eight months of twice-weekly supervised high-intensity progressive resistance and impact training (HiRIT) or supervised machine-based isometric axial compression (IAC) exercise training. Intervention group outcomes were compared at baseline and eight months with a matched but non-randomised control group (CON) who self-selected to usual activities. DXA scans (Medix DR, Medilink, France) of the skeletally non-dominant proximal femur were analysed using 3D hip software (DMS Group, France) to derive femoral neck (FN) and total hip (TH) bone mineral content (BMC), volume, and volumetric bone mineral density (vBMD) for total, trabecular and cortical bone compartments. Total FN cortical thickness was determined as well as anterior, posterior, lateral and medial subregions. pQCT scans (XCT-3000, Stratec, Germany) of the 4 and 38% sites of the tibia, and 4 and 66% sites of the radius were conducted to determine a range of geometric and bone structural strength indices. Intervention effects were examined using univariate ANCOVA of percent change, and repeated measures ANCOVA of raw baseline and follow-up data, controlling for initial values, using intention-to-treat and per-protocol approaches. RESULTS: Ninety-three men (67.1 ± 7.5 yrs, 175.2 ± 6.7 cm, 82.1 ± 11.6 kg, 26.7 ± 3.5 kg/m2) with lower than average aBMD (LS T-score -0.06 ± 1.04, FN T-score -1.58 ± 0.58, TH T-score -1.00 ± 0.58) were recruited, and designated CON (n = 26) or randomised to HiRIT (n = 34) or IAC (n = 33). Compliance to the supervised exercise programs did not differ (HiRIT 77.8 ± 16.6% versus IAC 78.5 ± 14.8%, p = 0.872). HiRIT improved medial FN cortical thickness compared with CON (5.6 ± 1.7% versus -0.1 ± 1.9%, p = 0.028) and IAC (5.6 ± 1.7% versus 0.7 ± 1.7%, p = 0.044). Distal tibia total BMC, vBMD, area and bone strength index, and trabecular BMC and bone strength index all declined for CON compared with maintenance for both HiRIT and IAC (all p < 0.05). HiRIT maintained distal tibia trabecular area compared with a loss in CON (0.2 ± 0.5% versus -1.6 ± 0.5%, p = 0.013). HiRIT and IAC maintained distal radius total BMC compared with loss in CON (-0.1 ± 0.7% versus -3.7 ± 0.8%, p = 0.001; 1.3 ± 0.7% versus -3.7 ± 0.8%, p < 0.001, respectively). HiRIT and IAC maintained distal radius total bone strength index compared with loss in CON (1.4 ± 1.4% versus -6.0 ± 1.6%, p = 0.001; 0.2 ± 1.3% versus -6.0 ± 1.6%, p = 0.004, respectively). HiRIT reduced proximal radius cortical area compared with CON (-3.1 ± 1.0% versus 1.1 ± 1.2%, p = 0.011) and IAC (-3.1 ± 1.0% versus -0.2 ± 1.0%, p = 0.042). No between-group differences were detected in any pQCT-derived bone outcome at the diaphyseal tibia 38% site. CONCLUSION: Findings indicate that supervised HiRIT provides a positive stimulus to cortical bone at the medial FN compared with supervised IAC exercise, and both HiRIT and IAC preserve bone strength at the distal tibia and distal radius. These effects may translate into a reduced risk of lower and upper extremity fracture in middle-aged and older men with low bone mass.

Multiscale modelling of drug transport and metabolism in liver spheroids.

In early preclinical drug development, potential candidates are tested in the laboratory using isolated cells. These in vitro experiments traditionally involve cells cultured in a two-dimensional monolayer environment. However, cells cultured in three-dimensional spheroid systems have been shown to more closely resemble the functionality and morphology of cells in vivo. While the increasing usage of hepatic spheroid cultures allows for more relevant experimentation in a more realistic biological environment, the underlying physical processes of drug transport, uptake and metabolism contributing to the spatial distribution of drugs in these spheroids remain poorly understood. The development of a multiscale mathematical modelling framework describing the spatio-temporal dynamics of drugs in multicellular environments enables mechanistic insight into the behaviour of these systems. Here, our analysis of cell membrane permeation and porosity throughout the spheroid reveals the impact of these properties on drug penetration, with maximal disparity between zonal metabolism rates occurring for drugs of intermediate lipophilicity. Our research shows how mathematical models can be used to simulate the activity and transport of drugs in hepatic spheroids and in principle any organoid, with the ultimate aim of better informing experimentalists on how to regulate dosing and culture conditions to more effectively optimize drug delivery.

A Comparison of Bone-Targeted Exercise Strategies to Reduce Fracture Risk in Middle-Aged and Older Men with Osteopenia and Osteoporosis: LIFTMOR-M Semi-Randomized Controlled Trial.

The Lifting Intervention For Training Muscle and Osteoporosis Rehabilitation for Men (LIFTMOR-M) trial examined efficacy and safety of two novel exercise programs in older men with low BMD. Men with low hip and/or LS BMD were randomized to high-intensity progressive resistance and impact training (HiRIT) or machine-based isometric axial compression (IAC) and compared to a nonrandomized matched control (CON). Outcomes included: hip and LS BMD; calcaneal ultrasound parameters; anthropometry; body composition; function (timed up-and-go [TUG], five-times sit-to-stand [FTSTS]); back extensor strength (BES); leg extensor strength (LES); compliance and adverse events. Ninety-three men (67.1 ± 7.5 years; 82.1 ± 11.6 kg; 175.2 ± 6.7 cm; FN T-score -1.6 ± 0.6) were randomized to HiRIT (n = 34) or IAC (n = 33), or allocated to CON (n = 26). HiRIT improved trochanteric BMD (2.8 ± 0.8%; -0.1 ± 0.9%, p = .024), LS BMD (4.1 ± 0.7%; 0.9 ± 0.8%, p = .003), BUA (2.2 ± 0.7%; -0.8 ± 0.9%, p = .009), stiffness index (1.6 ± 0.9%; -2.0 ± 1.1%, p = .011), lean mass (1.5 ± 0.8%; -2.4 ± 0.9%, p = .002), TUG, FTSTS, BES, and LES (p < .05) compared with CON. IAC improved lean mass (0.8 ± 0.8%; -2.4 ± 0.9%, p = .013) and FTSTS (-4.5 ± 1.6%; 7.5 ± 2.0%, p < .001) compared with CON. HiRIT improved LS BMD (4.1 ± 0.7%; 2.0 ± 0.7%, p = .039), stiffness index (1.6 ± 0.9%; -1.3 ± 0.9%, p = .025), and FTSTS (-10.7 ± 1.6%; -4.5 ± 1.7%, p = .010) compared with IAC. Exercise compliance was high (HiRIT 77.8 ± 16.6%; IAC 78.5 ± 14.8%, p = .872). There were five minor adverse events (HiRIT, 2; IAC, 3). HiRIT was well-tolerated and improved bone, function and fracture risk more than CON or IAC. © 2020 American Society for Bone and Mineral Research.

Enjoyment and acceptability of different exercise modalities to improve bone health in young adult women.

ISSUE ADDRESSED: Osteoporosis presents a serious public health issue and physical activity is recognised as the most effective modifiable risk factor for the condition. The reasons behind physical activity participation, however, are complex. We therefore aimed to explore the experiences related to a bone-targeted exercise intervention, determine enjoyment and acceptability of each exercise mode, and identify barriers and facilitators to osteogenic exercise for young adult women. METHODS: The present study was conducted within the Osteoporosis Prevention Through Impact and Muscle-loading Approaches to Exercise (OPTIMA-Ex) trial, a three-arm RCT comparing musculoskeletal outcomes from two supervised, high-intensity, exercise programs (impact and resistance training) with an unsupervised low-intensity exercise control. A mixed-methods approach was used, including quality of life and physical activity enjoyment questionnaires and qualitative analysis of semi-structured interviews. RESULTS: All groups had improvements in the 'mental health' domain of the quality of life measure; however, the two supervised exercise groups had greater levels of physical activity enjoyment. The qualitative analysis revealed that overall the trial activities were positively, yet the two supervised groups had 'richer' exercise experiences. Motivations for participation, barriers to physical activity and desired continuation of participation differed between all three groups. CONCLUSIONS: Findings suggest that bone-targeted exercise interventions for young adult women must address perceived time demands and environmental barriers to participation in order to maximise compliance and adherence. SO WHAT?: As physical activity is the most effective lifestyle strategy to improve bone health and young adulthood an important window for its augmentation, increasing convenience, accessibility and understanding of osteoporosis preventative behaviours in this demographic is vital.

Regional changes in indices of bone strength of upper and lower limbs in response to high-intensity impact loading or high-intensity resistance training.

It is well known that the bone response to physical activity is highly dependent on the nature of the loads imposed. Despite this, few direct comparisons of the effect of impact-style loading and resistance training on bone have been made. We therefore aimed to compare the effects of 10-month, twice-weekly, high-impact loading and 10-month, twice-weekly, high-intensity resistance training on indices of bone strength of both the upper and lower limbs of young adult women. Physically inactive, otherwise healthy, young adult women (18-30 years) with below average bone mass (T-score ≤ 0) were recruited as part of the OPTIMA-Ex trial. Testing included DXA- and pQCT-derived measures of bone mass and indices of bone strength and QUS-derived measures of bone quality of the dominant (D) and non-dominant (ND) upper (radius) and lower limbs (femoral neck, tibia, calcaneus). The present study examined those participants who completed the impact training (IT; n = 10) and resistance training (RT; n = 12) arms of the trial. Age differed between groups at baseline (IT = 23.2 ± 3.8 years, RT = 20.5 ± 1.8 years; p = 0.042). Compliance with the training programs did not differ (IT = 61.4 ± 15.1%, RT = 66.4 ± 11.2%, p = 0.381). Age and baseline differences in bone outcomes served as covariates for repeated measures and univariate ANCOVA conducted for dependent variables and percent change respectively. IT improved distal pQCT-derived bone mineral density (BMD) of the upper limb (ND radius: total BMD = 8.55 ± 2.26% versus 1.50 ± 2.04%, p = 0.040 and trabecular BMD = 1.86 ± 0.90% versus -1.30 ± 0.81%, p = 0.029) and lower limb (ND tibia trabecular BMD = 1.22 ± 0.55% versus -0.82 ± 0.50%, p = 0.017), more than RT. IT also improved upper limb bone strength index (BSI) (ND radius total BSI = 15.35 ± 2.83% versus 2.67 ± 2.55, p = 0.005) and lower limb BSI (D tibia total BSI = 5.16 ± 1.13% versus 0.37 ± 1.02%, p = 0.008; D tibia trabecular BSI = 3.93 ± 1.76% versus -2.84 ± 1.59, p = 0.014, ND tibia trabecular BSI = 3.57 ± 1.63% versus -3.15 ± 1.48%, p = 0.009) more than RT. Conversely, RT improved DXA-derived cortical volumetric BMD at the femoral neck more than IT (3.68 ± 1.99% versus -4.14 ± 2.20%, p = 0.021). Results suggest that IT and RT provide differing site-specific effects in both the upper and lower limbs, with superior bone responses observed at the distal segment from IT, while RT appeared to have greater effect on the shaft of the bone, on indices of bone-strength in young adult women.

Haptoglobin expression in human colorectal cancer.

Aims and experimental design. The acute-phase protein haptoglobin (Hp) has been recently detected in colorectal cancer (CRC) tissue, where its expression correlates with metastasis. Recently, we identified Hp as a CDw75 antigen-expressing protein in colorectal tissue. To deepen the knowledge of this protein in CRC, we studied the expression of Hp in healthy and tumour tissue specimens from 62 CRC patients by immunohistochemistry and Western blotting, as well as in the Caco-2 and HT-29 CRC cell lines by quantitative PCR, immunofluorescence microscopy and flow cytometry. Results and discussion. Hp immuno-positive staining was absent in the 18 healthy colorectal specimens analysed, whereas it was observed in 24% (15/62) of the tumour specimens as cytoplasmic granules within cancer cells. Furthermore, Hp expression in CRC was associated with Dukes' stage and the presence of metastasis in our population of study. In vitro cultured Caco-2 and HT-29 cells expressed mRNA for Hp and the protein was detected at the cell surface. Conclusions. This study confirms the expression of Hp in CRC, both in vivo and in vitro, and provides further evidence of its association with disease progression and metastasis.

Characterisation of a functional rat hepatocyte spheroid model.

Many in vitro liver cell models, such as 2D systems, that are used to assess the hepatotoxic potential of xenobiotics suffer major limitations arising from a lack of preservation of physiological phenotype and metabolic competence. To circumvent some of these limitations there has been increased focus on producing more representative 3D models. Here we have used a novel approach to construct a size-controllable 3D hepatic spheroid model using freshly isolated primary rat hepatocytes (PRH) utilising the liquid-overlay technique whereby PRH spontaneously self-assemble in to 3D microtissues. This system produces viable spheroids with a compact in vivo-like structure for up to 21 days with sustained albumin production for the duration of the culture period. F-actin was seen throughout the spheroid body and P-glycoprotein (P-gp) and multidrug resistance-associated protein 2 (MRP2) transporters had polarised expression on the canalicular membrane of hepatocytes within the spheroids upon formation (day 3). The MRP2 transporter was able to functionally transport 5 µM 5-chloromethylfluorescein diacetate (CMFDA) substrates into these canalicular structures. These PRH spheroids display in vivo characteristics including direct cell-cell contacts, cellular polarisation, 3D cellular morphology, and formation of functional secondary structures throughout the spheroid. Such a well-characterised system could be readily exploited for pre-clinical and non-clinical repeat-dose investigations and could make a significant contribution to replace, reduce and refine the use of animals for applied research.

High-Intensity Resistance and Impact Training Improves Bone Mineral Density and Physical Function in Postmenopausal Women With Osteopenia and Osteoporosis: The LIFTMOR Randomized Controlled Trial.

Optimal osteogenic mechanical loading requires the application of high-magnitude strains at high rates. High-intensity resistance and impact training (HiRIT) applies such loads but is not traditionally recommended for individuals with osteoporosis because of a perceived high risk of fracture. The purpose of the LIFTMOR trial was to determine the efficacy and to monitor adverse events of HiRIT to reduce parameters of risk for fracture in postmenopausal women with low bone mass. Postmenopausal women with low bone mass (T-score < -1.0, screened for conditions and medications that influence bone and physical function) were recruited and randomized to either 8 months of twice-weekly, 30-minute, supervised HiRIT (5 sets of 5 repetitions, >85% 1 repetition maximum) or a home-based, low-intensity exercise program (CON). Pre- and post-intervention testing included lumbar spine and proximal femur bone mineral density (BMD) and measures of functional performance (timed up-and-go, functional reach, 5 times sit-to-stand, back and leg strength). A total of 101 women (aged 65 ± 5 years, 161.8 ± 5.9 cm, 63.1 ± 10.4 kg) participated in the trial. HiRIT (n = 49) effects were superior to CON (n = 52) for lumbar spine (LS) BMD (2.9 ± 2.8% versus -1.2 ± 2.8%, p < 0.001), femoral neck (FN) BMD (0.3 ± 2.6% versus -1.9 ± 2.6%, p = 0.004), FN cortical thickness (13.6 ± 16.6% versus 6.3 ± 16.6%, p = 0.014), height (0.2 ± 0.5 cm versus -0.2 ± 0.5 cm, p = 0.004), and all functional performance measures (p < 0.001). Compliance was high (HiRIT 92 ± 11%; CON 85 ± 24%) in both groups, with only one adverse event reported (HiRIT: minor lower back spasm, 2/70 missed training sessions). Our novel, brief HiRIT program enhances indices of bone strength and functional performance in postmenopausal women with low bone mass. Contrary to current opinion, HiRIT was efficacious and induced no adverse events under highly supervised conditions for our sample of otherwise healthy postmenopausal women with low to very low bone mass. © 2017 American Society for Bone and Mineral Research.