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Bisphosphonates limit resorption by inhibiting osteoclast formation and activation. They are removed during preparation of demineralized bone matrix (DBM) particles, but it is not known if osteogenesis and incorporation of mineralized bone allografts from patients treated with oral bisphosphonates are affected in vivo.Human block allografts from 3 bisphosphonate-treated donors and 3 age and sex-matched control donors who had not received bisphosphonates were obtained (Musculoskeletal Transplant Foundation); one-half from each donor was demineralized. In the first study, 3 × 2-mm mineralized and demineralized cylindrical grafts were implanted bilaterally in the femoral metaphysis of 56 rats. In the second study, samples from each group were pooled, prepared as particles, and implanted bilaterally in the femoral marrow canal of 24 rats. Osseointegration, defined as native bone in contact with allograft, was assessed at 10 weeks by micro-computed tomography (CT) and histomorphometry.Micro-CT showed greater bone volume in sites treated with demineralized samples compared with the control mineralized and bisphosphonate-exposed mineralized samples. More new bone was generated along the cortical-endosteal interface compared with mineralized samples. Histology showed significantly less new bone in contact with the mineralized bisphosphonate-exposed allograft (10.4%) compared with mineralized samples that did not receive bisphosphonates (22.8%) and demineralized samples (31.7% and 42.8%). A gap was observed between native bone and allograft in the bisphosphonate-exposed mineralized samples (0.50 mm 2 ). The gap area was significantly greater compared with mineralized samples that did not receive bisphosphonates (0.16 mm 2 ) and demineralized samples (0.10 and 0.03 mm 2 ).Mineralized allografts were osseointegrated, but not remodeled or replaced by living bone, preventing full regeneration of the bone defect. Prior treatment of the donor with bisphosphonates affected osteogenesis, preventing osteointegration and remodeling of the allograft into the regenerating bone.Clinical use of mineralized allografts from patients who had received bisphosphonate therapy needs to be evaluated; in this animal model, such grafts were not integrated into the host bone or remodeled, and full regeneration of the bone defects was prevented.

Supplemental Table 1: Gene expression primers used in quantitative real-time PCR. Supplemental Table 2: Antibodies used in western blots. Figure S1: The effect of 24R,25(OH)2D3 on MDA-MB-231 cells. Figure S2: The effect of 24R,25(OH)2D3 on T-47D cells. Figure S3: Silencing CAV1 in HCC38 cells. Figure S4: Whole western blot of silencing of caveolin-1 in HCC38 cells from Supplemental Figure S3. Figure S5: Overexpression of ESR1 in HCC38 changes ERα isoform expression. Figure S6: Whole western blot of transient overexpression of ESR1 in HCC38 cells from Supplemental Figure S5. Figure S7: Overexpression of ESR1 in C2C12. Figure S8: Whole western blot of stable overexpression of ESR1 in C2C12 cells from Supplemental Figure 7. Figure S9: ERα66 was stably knocked down in MCF7 using a recombinant plasmid with a puromycin antibiotic resistant selection marker. Figure S10: Whole western blot of stable silencing of ERα66 in MCF7 cells from Supplemental Figure 9. Figure S11: Silencing the ERα66 isoform reversed the pro-apoptotic effect of 24R,25(OH)2D3 on MCF7 cells.

Purpose:Advanced-stage mucinous ovarian carcinoma (MOC) has poor chemotherapy response and prognosis and lacks biomarkers to aid stage I adjuvant treatment. Differentiating primary MOC from gastrointestinal (GI) metastases to the ovary is also challenging due to phenotypic similarities. Clinicopathologic and gene-expression data were analyzed to identify prognostic and diagnostic features.Experimental Design:Discovery analyses selected 19 genes with prognostic/diagnostic potential. Validation was performed through the Ovarian Tumor Tissue Analysis consortium and GI cancer biobanks comprising 604 patients with MOC (n = 333), mucinous borderline ovarian tumors (MBOT, n = 151), and upper GI (n = 65) and lower GI tumors (n = 55).Results:Infiltrative pattern of invasion was associated with decreased overall survival (OS) within 2 years from diagnosis, compared with expansile pattern in stage I MOC [hazard ratio (HR), 2.77; 95% confidence interval (CI), 1.04–7.41, P = 0.042]. Increased expression of THBS2 and TAGLN was associated with shorter OS in MOC patients (HR, 1.25; 95% CI, 1.04–1.51, P = 0.016) and (HR, 1.21; 95% CI, 1.01–1.45, P = 0.043), respectively. ERBB2 (HER2) amplification or high mRNA expression was evident in 64 of 243 (26%) of MOCs, but only 8 of 243 (3%) were also infiltrative (4/39, 10%) or stage III/IV (4/31, 13%).Conclusions:An infiltrative growth pattern infers poor prognosis within 2 years from diagnosis and may help select stage I patients for adjuvant therapy. High expression of THBS2 and TAGLN in MOC confers an adverse prognosis and is upregulated in the infiltrative subtype, which warrants further investigation. Anti-HER2 therapy should be investigated in a subset of patients. MOC samples clustered with upper GI, yet markers to differentiate these entities remain elusive, suggesting similar underlying biology and shared treatment strategies.

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Takotsubo cardiomyopathy (TCM), or stress-induced cardiomyopathy, is associated with adverse prognosis. Limited data suggest that TCM occurring in orthotopic liver transplant (OLT) recipients is associated with elevated peri-operative risk.To characterize the predictors of TCM in OLT recipients, using a large, multi-center pooled electronic health database.A multi-institutional database (Explorys Inc, Cleveland, OH, USA), an aggregate of de-identified electronic health record data from 26 United States healthcare systems was surveyed. A cohort of patients with a Systematized Nomenclature of Medicine-Clinical Terms of "liver transplant" between 09/2015 and 09/2020 was identified. Subsequently, individuals who developed a new diagnosis of TCM following OLT were identified. Furthermore, the risk associations with TCM among this patient population were characterized using linear regression.Between 09/2015 and 09/2020, of 37718540 patients in the database, 38740 (0.10%) had a history of OLT (60.6% had an age between 18-65 years, 58.1% female). A new diagnosis of TCM was identified in 0.3% of OLT recipients (45.5% had an age between 18-65 years, 72.7% female), compared to 0.04% in non-OLT patients [odds ratio (OR): 7.98, 95% confidence intervals: 6.62-9.63, (TCM was significantly more likely to occur in LT recipients

Purpose: Ovarian cancer recurrence with the development of paclitaxel resistance is an obstacle to long-term survival. We showed that secretory leukocyte protease inhibitor (SLPI) is a survival factor for ovarian cancer. We hypothesize that SLPI may antagonize paclitaxel injury.Experimental Design: Differential SLPI induction in response to paclitaxel and in response to stable forced expression of SLPI was shown in A2780-1A9 cells and their paclitaxel-resistant sublines, PTX10 and PTX22, and confirmed with HEY-A8 cells. SLPI-mediated survival was reduced by the MAP/extracellular signal-regulated kinase (ERK) kinase inhibitor, U0126, and a humanized neutralizing monoclonal anti-SLPI antibody, CR012. OVCAR3 xenographs tested the role of CR012 in vivo.Results: SLPI expression was lower in A2780-1A9 ovarian cancer cells than in PTX10 and PTX22, and SLPI was induced by paclitaxel exposure. Stable SLPI expression yielded a proliferation advantage (P = 0.01); expression of and response to SLPI in OVCAR3 cells were abrogated by exposure to CR012. SLPI reduced the paclitaxel susceptibility of 1A9 and HEY-A8 cells (P ≤ 0.05), and SLPI expression did not increase the resistance of PTX10 and PTX22 cells. Both paclitaxel and SLPI overexpression induced ERK activation. Inhibition of MAP/ERK kinase with U0126 increased paclitaxel injury and overcame SLPI-mediated cell protection. It did not reinstate PTX10 sensitivity to paclitaxel, which was associated with AKT activation. Significant inhibition of OVCAR3 xenograft growth was observed with CR012 and paclitaxel, over single agents (P ≤ 0.001).Conclusions: A two-pronged approach confirmed that SLPI overcomes paclitaxel in part through activation of ERK1/2. These results credential SLPI as a molecular target for ovarian cancer and suggest CR012 as a tool for proof of concept. Clin Cancer Res; 16(2); 600–9

Reduced skeletal loading associated with many conditions, such as neuromuscular injuries, can lead to bone fragility and may threaten the success of implant therapy. Our group has developed a botulinum toxin A (botox) injection model to imitate disease-reduced skeletal loading and reported that botox dramatically impaired the bone formation and osseointegration of titanium implants. Semaphorin 3A (sema3A) is an osteoprotective factor that increases bone formation and inhibits bone resorption, indicating its potential therapeutic role in improving osseointegration in vivo. We first evaluated the sema3A effect on whole bone morphology following botox injections by delivering sema3A via injection. We then evaluated the sema3A effect on the osseointegration of titanium implants with two different surface topographies by delivering sema3A to cortical bone defect sites prepared for implant insertion and above the implants after insertion using a copper-free click hydrogel that polymerizes rapidly in situ. Implants had hydrophobic smooth surfaces (PT) or multiscale biomimetic micro/nano topography (SLAnano). Sema3A rescued the botox-impaired bone formation. Furthermore, biomimetic Ti implants improved the bone-to-implant contact (BIC) and mechanical properties of the integrated bone in the botox-treated rats, which sema3A enhanced. This study demonstrated the value of biomimetic approaches combining multiscale topography and biologics in improving the clinical outcomes of implant therapy.

This article describes a teaching plan for a discussion-driven introduction to moral reasoning and explains its philosophical and pedagogical rationale. The teaching plan consists of a sequence of ...

Emotional responses arise from limbic circuits including the hippocampus and amygdala. In the human brain, beta-frequency communication between these structures correlates with self-reported mood and anxiety. However, both the mechanism and significance of this biomarker as a readout vs. driver of emotional state remain unknown. Here we show that beta-frequency communication between the ventral hippocampus and basolateral amygdala also predicts anxiety-related behavior in mice on both long timescales (∼30 min) and immediately preceding behavioral choices. Genetically encoded voltage indicators reveal that this biomarker reflects synchronization between somatostatin interneurons across both structures. Indeed, synchrony between these neurons dynamically predicts approach vs. avoidance, and optogenetically shifting this synchronization by just 25 msec is sufficient to bidirectionally modulate anxiety-related behaviors. Thus, back-translation establishes a human biomarker as a causal determinant (not just predictor) of emotional state, revealing a novel mechanism whereby interregional synchronization that is frequency-, phase- and cell type-specific controls anxiety processing.

Purpose This study aims to discuss science governance in Europe and the network of associated nonprofit institutions. The authors posit that this network, which comprises both (partial) learning organizations and non-learning organizations, has been observed to postpone taking up “responsibility” as an issue in science governance and funding decisions. Design/methodology/approach This paper discusses the challenge of learning and policy implementation within the European science governance system. By exploring how learning on responsible innovation (RI) in this governance system can be provoked, it addresses the question how Senge’s insights in organizational learning can clarify discourses on and practices of RI and responsibility in research. This study explores the potential of a new organizational form, that of Social Labs, to support learning on Responsible Research and Innovation (RRI) in standing governance organizations. Findings This study concludes that Social Labs are a suitable format for enacting the five disciplines as identified by Senge, and a Social Lab may turn into a learning organization, be it a temporary one. Responsibility in research and innovation is conducive for learning in the setting of a Social Lab, and Social Labs act as intermediary organizations, which not merely pass on information among actors but also actively give substantive shape to what they convey from a practice-informed, normative orientation. Research limitations/implications This empirical work on RRI-oriented Social Labs therefore suggests that Social Lab–oriented temporary, intermediary learning organizations present a promising form for implementing complex normative policies in a networked, nonhierarchical governance setting. Practical implications Based on this research funding and governance organizations in research, policy-makers in other domains may take up and create such intermediary organizations to aid learning in (science) governance. Social implications This research suggests that RRI-oriented Social Labs present a promising form for implementing complex normative policies, thus integrate learning on and by responsible practices in various governance settings. Originality/value European science governance is characterized by a network of partial Learning Organization (LOs) and Non-Learning Organization (nLOs) who postpone decision-making on topics around “responsibility” and “solving societal challenges” or delegate authority to reviewers and individual actors, filtering possibilities for collaborative transformation toward RRI. social lab (SLs) are spaces that can address social problems or social challenges in an open, action-oriented and creative manner. As such, they may function as temporary, intermediary LOs bringing together diverse actors from a specific context to work on and learn about issues of science and society where standing organizations avoid doing so. Taken together, SLs may offer temporary organizational structures and spaces to move beyond top-down exercise of power or lack of real change to more open, deliberative and creative forms of sociopolitical coordination between multiple actors cutting across realms of state, practitioners of research and innovation and civil society. By taking the role of temporary LOs, they may support existing research and innovation organizations and research governance to become more flexible and adaptive.

Objective Modifications to implant surface properties, including topography, chemistry, and wettability, alter immune response, osteoblast differentiation of bone marrow stromal cells (MSCs), and implant integration in vivo. Dielectric barrier discharge (DBD) plasma treatment has been used to sterilize surfaces and remove adsorbed carbon, improving wettability. However, unless it is used immediately prior to placement, ambient atmospheric hydrocarbons rapidly adhere to the surface, thereby reducing its hydrophilicity. Moreover, this method is not practical in many clinical settings. The aim of this study was to evaluate the effectiveness of an on-site benchtop modification technique for implants at time of placement, consisting of a DBD plasma that is used to sterilize implants that are pre-packaged in a vacuum. Effects of the plasma-treatment on implant surface properties and cellular response of MSCs and osteoblasts were assessed in vitro. Methods Titanium-aluminum-vanadium implant surfaces were grit-blasted (GB) or grit-blasted and acid-etched (AE), and packaged under vacuum. AE surfaces were also plasma-treated using the benchtop device (GB + AE) and then removed from the vacuum. GB surface morphology was altered with AE but AE microroughness was not changed with the plasma-treatment. Plasma-treatment increased the surface wettability, but did not alter surface atomic concentrations of titanium, oxygen, or carbon. Results MSCs and osteoblast-like cells (MG63 s) produced increased concentrations of osteocalcin, osteopontin, and osteoprotegerin after plasma-treatment of AE surfaces compared to non-plasma-treated AE surfaces; production of IL6 was reduced and IL10 was. Aging GB + AE surfaces for 7 days after plasma-treatment but still in the vacuum environment reduced the effectiveness of plasma on cellular response. Significance Overall, these data suggest that application of benchtop plasma at the time of implant placement can alter the surface free energy of an implant surface without modifying surface chemical composition and enhance the differentiation and activity of MSCs and osteoblasts that are in contact with these implant surfaces.

Transfection of chondrocytes with microRNA-451(miR-451), present in growth zone cartilage of the growth plate, upregulates production of enzymes association with extracellular matrix degradation. miR-451 is also present in articular cartilage and exacerbates IL-1β effects in articular chondrocytes. Moreover, when osteoarthritis (OA) was induced in Sprague Dawley rats via bilateral anterior cruciate ligament transection (ACLT), miR-451 expression was increased in OA cartilage compared to control, suggesting its inhibition might be used to prevent or treat OA. To examine the prophylactic and therapeutic potential of inhibiting miR-451, we evaluated treatment with miR-451 power inhibitor (451-PI) at the onset of joint trauma and treatment after OA had developed. The prophylactic animal cohort received twice-weekly intra-articular injections of either 451-PI or a negative control (NC-PI) beginning on post-surgical day 3. OA was allowed to develop for 24 days in the therapeutic cohort before beginning injections. All rats were killed on day 45. Micro-CT, histomorphometrics, OARSI scoring, and muscle force testing were performed on samples. 451-PI mitigated OA progression compared to NC-PI limbs in the prophylactic cohort based on histomorphometric analysis and OARSI scoring, but no differences were detected by micro-CT. 451-PI treatment beginning 24 days post-surgery was not able to reduce OA severity. Prophylactic administration of 451-PI mitigates OA progression in a post-trauma ACLT rat model supporting its potential to prevent OA development following an ACLT injury clinically.

An aging global population is accelerating the need for better, longer-lasting orthopaedic and dental implants. Additive manufacturing can provide patient-specific, titanium-alloy-based implants with tailored, three-dimensional, bone-like architecture. Studies using two-dimensional substrates have demonstrated that osteoblastic differentiation of bone marrow stromal cells (MSCs) is enhanced on surfaces possessing hierarchical macro/micro/nano-scale roughness that mimics the topography of osteoclast resorption pits on the bone surface. Conventional machined implants with these surfaces exhibit successful osseointegration, but the complex architectures produced by 3D printing make consistent nanoscale surface texturing difficult to achieve, and current line-of-sight methods used to roughen titanium alloy surfaces cannot reach all internal surfaces. Here, we demonstrate a new, non-line-of-sight, gas/solid-reaction-based process capable of generating well-controlled nanotopographies on all open (gas-exposed) surfaces of titanium alloy implants. Dense 3D-printed titanium-aluminum-vanadium (TiAl6V4) substrates were used to evaluate the evolution of surface nanostructure for development of this process. Substrates were either polished to be smooth (for easier evaluation of surface nanostructure evolution) or grit-blasted and acid-etched to present a microrough biomimetic topography. An ultrathin (90 ± 16 nm) conformal, titania-based surface layer was first formed by thermal oxidation (600 °C, 6 h, air). A calciothermic reduction (CaR) reaction (700 °C, 1 h) was then used to convert the surface titania (TiO2) into thin layers of calcia (CaO, 77 ± 16 nm) and titanium (Ti, 51 ± 20 nm). Selective dissolution of the CaO layer (3 M acetic acid, 40 min) then yielded a thin nanoporous/nanorough Ti-based surface layer. The changes in surface nanostructure/chemistry after each step were confirmed by scanning and transmission electron microscopies with energy-dispersive X-ray analysis, X-ray diffraction, selected area electron diffraction, atomic force microscopy, and mass change analyses. In vitro studies indicated that human MSCs on CaR-modified microrough surfaces exhibited increased protein expression associated with osteoblast differentiation and promoted osteogenesis compared to unmodified microrough surfaces (increases of 387% in osteopontin, 210% in osteocalcin, 282% in bone morphogenic protein 2, 150% in bone morphogenic protein 4, 265% in osteoprotegerin, and 191% in vascular endothelial growth factor). This work suggests that this CaR-based technique can provide biomimetic topography on all biologically facing surfaces of complex, porous, additively manufactured TiAl6V4 implants.

Vitamin D3 and its metabolites have antitumorigenic properties in vitro and in vivo; however, clinical trials and retrospective studies on the effectiveness of vitamin D3 oral supplementation against cancer have been inconclusive. One reason for this may be that clinical trials ignore the complex vitamin D metabolome and the many active vitamin D3 metabolites present in the body. Recent work by our lab showed that 24R,25(OH)2D3, a vitamin D3 metabolite that is active in chondrocyte proliferation and differentiation, has antitumorigenic properties in estrogen receptor alpha-66 (ERα66)–positive (ER+) breast cancer, but not in ERα66-negative (ER−) breast cancer. Here we show that 24R,25(OH)2D3 is protumorigenic in an in vivo mouse model (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice) of ER− breast cancer, causing greater tumor growth than in mice treated with vehicle alone. In vitro results indicate that the effect of 24R,25(OH)2D3 is via a membrane-associated mechanism involving ERs and phospholipase D. 24R,25(OH)2D3 increased proliferation and reduced apoptosis in ERα66-negative HCC38 breast cancer cells, and stimulated expression of metastatic markers. Overexpressing ESRI, which encodes ERα66, ERα46, and ERα36, reduced the proapoptotic response of ERα66− cells to 24R,25(OH)2D3, possibly by upregulating ERα66. Silencing ESR1 in ERα66+ cells increased apoptosis. This suggests 24R,25(OH)2D3 is differentially tumorigenic in cancers with different ERα isoform profiles. Antiapoptotic actions of 24R,25(OH)2D3 require ERα36 and proapoptotic actions require ERα66. Implications: These results suggest that 24R,25(OH)2D3, which is a major circulating metabolite of vitamin D, is functionally active in breast cancer and that the regulatory properties of 24R,25(OH)2D3 are dependent upon the relative expression of ERα66 and ERα36.

This article considers the extent to which the Holocaust galvanized British antifascism in the 1960s. It explores whether the genocide surfaced in Jewish antifascists’ motivations and rhetoric but goes beyond this to assess the Holocaust’s political capital in wider antifascism and anti-racism. The article considers whether political coalitions were negotiated around Holocaust memory, for example, by analysing whether Jewish antifascism intersected with the black and Asian communities of Smethwick and Southall respectively who were targeted by the far right in 1964. Using archival materials and newly-collected oral histories, the article surveys organisations including the Jewish Board of Deputies, the 62 Group, Yellow Star Movement and Searchlight newspaper. It will argue that the Holocaust played a more important role in 1960s’antifascism than has been recognised. Jewish groups fragmented around the lessons of the genocide for their antifascism. The Holocaust influenced race relations legislation and became a metonym for extreme racist violence.

Objectives This study sought to investigate the association between nonalcoholic fatty liver disease (NAFLD) and arrhythmia recurrence following atrial fibrillation ablation; and to examine the impact of NAFLD stage on outcomes. Background Metabolic derangements, including obesity and diabetes, are associated with incident and recurrent atrial fibrillation (AF), in addition to the development of NAFLD. Methods This was a retrospective study of 267 consecutive patients undergoing AF ablation, 89 of whom were diagnosed with NAFLD prior to ablation and matched in a 2:1 manner based on age, sex, body mass index, ejection fraction, and AF type with 178 patients without NAFLD. Patients were monitored for arrhythmia recurrence during a mean follow-up of 29 months. Results Recurrent arrhythmia was observed in 50 (56%) patients with NAFLD compared with 37 (21%) without NAFLD. Epicardial fat volume was measured on computed tomography and was significantly higher among those with NAFLD (248 ± 125 ml vs. 223 ± 97 ml; p = 0.01). On multivariable models adjusting for sleep apnea, body mass index, heart failure, AF type, and left atrial size, NAFLD was independently associated with increased rates of arrhythmia recurrence (hazard ratio: 3.010; 95% confidence interval: 1.980 to 4.680; p Conclusions NAFLD is associated with significantly increased arrhythmia recurrence rates following AF ablation. Identification and reversal, where possible, may result in improved arrhythmia-free survival.

We evaluated the frequency of absence of the palmaris longus tendon in the heterogeneous Israeli population. Nine hundred and fifty wrists were evaluated using a modified Mishra/Schaeffer technique (thumb/little-finger opposition with resisted wrist flexion), which was validated by ultrasound scanning. The geographical and ethnic origin of volunteers was documented. When physical examination was equivocal, any vague, superficial structure was subsequently identified as the median nerve by ultrasound. Physical examination reliably identified palmaris longus only when a structure was clinically obvious (visually or by palpation). There was bilateral absence of the palmaris longus in 21% and unilateral absence in 15% of participants. Frequency of bilateral absence varied between 4.5% and 30%, depending on geographical origin ( p = 0.0007). The incidence of palmaris longus tendon varied significantly by geographical, but not by ethnic origin. Level of evidence: II

The analysis of cell-free DNA (cfDNA) from plasma offers great promise for the earlier detection of cancer. At present, changes in DNA sequence, methylation, or copy number are the most sensitive ways to detect the presence of cancer. To increase the sensitivity of such assays, it would be useful to be able evaluate the same template molecules for all these changes. Here we report an approach, called MethylSaferSeqS, that achieves this goal, and can be applied to any standard library preparation method suitable for massively parallel sequencing. The innovative step was to copy both strands of each DNA-barcoded molecule with a primer that allows the subsequent separation of the original strands (retaining their 5-methylcytosine residues) from the copied strands (in which the 5-methylcytosine residues are replaced with unmodified cytosine residues). The original and copied strands are queried for epigenetic and genetic alterations, respectively. We applied this approach to plasma from 265 individuals, including 198 with cancers of the pancreas, ovary, lung or colon, and found the expected patterns of mutations, copy number alterations, and methylation. Furthermore, we could determine which original template DNA molecules were methylated and/or mutated. MethylSaferSeqS should be useful for addressing a variety of questions relating genetics and epigenetics in the future. Citation Format: Yuxuan Wang, Christopher Douville, Joshua Cohen, Chetan Bettegowda, Nick Papadopoulos, Ken Kinzler, Bert Vogelstein. Detection of rare mutations, copy number variation, and DNA methylation in the same template DNA molecules. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6609.

BackgroundCoformulated sodium phenylbutyrate/taurursodiol (PB/TURSO) was shown to prolong survival and slow functional decline in amyotrophic lateral sclerosis (ALS).ObjectiveDetermine whether PB/TURSO prolonged tracheostomy/ventilation-free survival and/or reduced first hospitalisation in participants with ALS in the CENTAUR trial.MethodsAdults with El Escorial Definite ALS ≤18 months from symptom onset were randomised to PB/ TURSO or placebo for 6 months. Those completing randomised treatment could enrol in an open-label extension (OLE) phase and receive PB/TURSO for ≤30 months. Times to the following individual or combined key events were compared in the originally randomised treatment groups over a period spanning trial start through July 2020 (longest postrandomisation follow-up, 35 months): death, tracheostomy, permanent assisted ventilation (PAV) and first hospitalisation.ResultsRisk of any key event was 47% lower in those originally randomised to PB/TURSO (n=87) versus placebo (n=48, 71% of whom received delayed-start PB/TURSO in the OLE phase) (HR=0.53; 95% CI 0.35 to 0.81; p=0.003). Risks of death or tracheostomy/PAV (HR=0.51; 95% CI 0.32 to 0.84; p=0.007) and first hospitalisation (HR=0.56; 95% CI 0.34 to 0.95; p=0.03) were also decreased in those originally randomised to PB/TURSO.ConclusionsEarly PB/TURSO prolonged tracheostomy/PAV-free survival and delayed first hospitalisation in ALS.Trial registration numberNCT03127514;NCT03488524.

Optimal glycemic control is vital in decreasing the risk of congenital birth defects and perinatal complications in women with diabetes. Although frequent blood glucose (BG) monitoring is essential during pregnancy, studies have highlighted poor compliance and falsification of glucose readings. We designed this study to assess whether a web-based glucose monitor improves compliance, glycemic control, and patient satisfaction. This was a prospective study of 30 women with pre-gestational diabetes. After 4 weeks of using paper logs, patients were given a web-based glucose monitor. The primary outcome of interest was the average number of BG readings prior to and during web-based implementation. Secondary outcomes included glycemic control and patient satisfaction as determined by a pre- and post-study survey. The number of BG readings after 2 months using the web-based meter was similar to baseline. Hemoglobin A1c (HbA1c) significantly improved and there was a trend toward improved overall glycemic values. Survey results demonstrated satisfaction with the new system, although 20% of patients felt uncomfortable with glucose values being available to providers in real time. Compliance with BG monitoring was similar when comparing a web-based system with written logs. Since other studies have highlighted that some glucose data from written logs are falsified, actual compliance using the web-based monitor may be improved. This study demonstrates potential patient concerns using a web-based system. Further studies should explore patient reactions to providers having real-time access to online glycemic data given our survey results.

Osseointegration of titanium-based implants possessing complex macroscale/microscale/mesoscale/nanoscale (multiscale) topographies support a direct and functional connection with native bone tissue by promoting recruitment, attachment and osteoblastic differentiation of bone marrow stromal cells (MSCs). Recent studies show that the MSCs on these surfaces produce factors, including bone morphogenetic protein 2 (BMP2) that can cause MSCs not on the surface to undergo osteoblast differentiation, suggesting they may produce an osteogenic environment in vivo. This study examined if soluble factors produced by MSCs in contact with titanium–aluminum–vanadium (Ti6Al4V) implants possessing a complex multiscale biomimetic topography are able to induce osteogenesis ectopically. Ti6Al4V disks were grit-blasted and acid-etched to create surfaces possessing macroscale and microscale roughness (MM), micro/meso/nanoscale topography (MN), and macro/micro/meso/nanoscale topography (MMNTM). Polyether-ether-ketone (PEEK) disks were also fabricated by machining to medical-grade specifications. Surface properties were assessed by scanning electron microscopy, contact angle, optical profilometry, and x-ray photoelectron spectroscopy. MSCs were cultured in growth media (GM). Proteins and local factors in their conditioned media (CM) were measured on days 4, 8, 10 and 14: osteocalcin, osteopontin, osteoprotegerin, BMP2, BMP4, and cytokines interleukins 6, 4 and 10 (IL6, IL4, and IL10). CM was collected from D14 MSCs on MMNTM and tissue culture polystyrene (TCPS) and lyophilized. Gel capsules containing active demineralized bone matrix (DBM), heat-inactivated DBM (iDBM), and iDBM + MMN-GM were implanted bilaterally in the gastrocnemius of athymic nude mice (N = 8 capsules/group). Controls included iDBM + GM; iDBM + TCPS-CM from D5 to D10 MSCs; iDBM + MMN-CM from D5 to D10; and iDBM + rhBMP2 (R&D Systems) at a concentration similar to D5–D10 production of MSCs on MMNTM surfaces. Legs were harvested at 35D. Bone formation was assessed by micro computed tomography and histomorphometry (hematoxylin and eosin staining) with the histology scored according to ASTM 2529–13. DNA was greatest on PEEK at all time points; DNA was lowest on MN at early time points, but increased with time. Cells on PEEK exhibited small changes in differentiation with reduced production of BMP2. Osteoblast differentiation was greatest on the MN and MMNTM, reflecting increased production of BMP2 and BMP4. Pro-regenerative cytokines IL4 and IL10 were increased on Ti-based surfaces; IL6 was reduced compared to PEEK. None of the media from TCPS cultures was osteoinductive. However, MMN-CM exhibited increased bone formation compared to iDBM and iDBM + rhBMP2. Furthermore, exogenous rhBMP2 alone, at the concentration found in MMN-CM collected from D5 to D10 cultures, failed to induce new bone, indicating that other factors in the CM play a critical role in that osteoinductive microenvironment. MSCs cultured on MMNTM Ti6Al4V surfaces differentiate and produce an increase in local factors, including BMP2, and the CM from these cultures can induce ectopic bone formation compared to control groups, indicating that the increased bone formation arises from the local response by MSCs to a biomimetic, multiscale surface topography.

The end Pleistocene was a time of ecological turmoil, coincident with environmental change, extinctions, and anthropogenic impacts on the landscape. As one of the few persisting predators from the Pleistocene, La Brea’s exceptional record of coyotes (Canis latrans) provides a unique opportunity to clarify how a recently documented ecosystem state-shift impacted survivors. Through a multiproxy analysis of Rancho La Brea coyotes from the past 50,000 years to present, we analyzed over 100 individuals for radiocarbon chronologies, stable isotopes, dental microwear, and morphology to assess the consequences of megafaunal extirpation on these predators. Most notably, coyotes demonstrate a significant decline in δ15Nbone collagen values immediately after the extirpation of megafauna. While this decline is suggestive of a change in diet from more to less meat, stable isotopes of amino acids from a subset of samples instead provide evidence of a baseline shift in nitrogen—indicating large scale changes in the availability of nutritional resources. While coyotes do not demonstrate notable changes in diet across the extirpation boundary, as inferred from stable carbon isotopes in tooth enamel and dental microwear texture analysis, significant shifts in stable oxygen isotopes in δ18Oenamel and δ13Cbone collagen indicate more nuanced changes in potential prey-resources. Coyotes also demonstrate a linear decline in body size that begins prior to the local extirpation of megafauna (~20,000 years ago) and may be in response to competition with larger canids, the decline in large prey, and/or concurrent increases in aridity during this interval. A dramatic increase in scavenging of forested prey (e.g., deer) during the past century stands out as significantly distinct from the dietary niches occupied over the past 50,000 years—implying dramatic impacts of human behavior on coyotes, a recent shift in their ecological role, and the highly adaptable nature of these carnivores.