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Idiopathic osteoporosis (IOP) is a rare form of early-onset osteoporosis diagnosed in patients with no known metabolic or hormonal cause of bone loss and unknown pathogenesis. Patients with IOP commonly report both childhood fractures and family history of osteoporosis, raising the possibility of genetic etiologies of IOP. Whole-exome sequencing analyses of different IOP cohorts identified multiple variants in melatonin receptor 1A ( MTNR1A ) with a potential pathogenic outcome. A rare MTNR1A variant (rs374152717) was found in members of an Ashkenazi Jewish family with IOP, and an MTNR1A variant (rs28383653) was found in a nonrelated female IOP cohort (4%). Both variants occur at a substantially higher frequency in Ashkenazi Jewish individuals than in the general population. We investigated consequences of the heterozygous (rs374152717) variant [ MTNR1A c.184+1G>T ( MTNR1A c. 184+1G>T )] on bone physiology. A mouse model of the human rs374152717 variant reproduced the low bone mass (BM) phenotype of young-adult patients with IOP. Low BM occurred because of induction of senescence in mutant osteoblasts followed by compromised differentiation and function. In human cells, introduction of rs374152717 led to translation of a nonfunctional protein and subsequent dysregulation of melatonin signaling. These studies provide evidence that MTNR1A mutations entail a genetic etiology of IOP and establish the rs374152717 variant as a loss-of-function allele that impairs bone turnover by inducing senescence in osteoblasts. The higher prevalence of the MTNR1A variants identified in IOP cohorts versus the general population indicates a greater risk of IOP in those carrying these variants, especially Ashkenazi Jewish individuals bearing the rs374152717 variant.

Purpose of Review: Along with a strong impact on skeletal integrity, bone marrow adipose tissue (BMAT) is an important modulator of the adult hematopoietic system. This review will summarize the current knowledge on the causal relationship between bone marrow (BM) adipogenesis and the development and progression of hematologic malignancies., Recent Findings: BM adipocytes (BMAds) support a number of processes promoting oncogenesis, including the evolution of clonal hematopoiesis, malignant cell survival, proliferation, angiogenesis, and chemoresistance. In addition, leukemic cells manipulate surrounding BMAds by promoting lipolysis and release of free fatty acids, which are then utilized by leukemic cells via β-oxidation. Therefore, limiting BM adipogenesis, blocking BMAd-derived adipokines, or lipid metabolism obstruction have been considered as potential treatment options for hematological malignancies. Leukemic stem cells rely heavily on BMAds within the structural BM microenvironment for necessary signals which foster disease progression. Further development of 3D constructs resembling BMAT at different skeletal regions are critical to better understand these relationships in geometric space and may provide essential insight into the development of hematologic malignancies within the BM niche. In turn, these mechanisms provide promising potential as novel approaches to targeting the microenvironment with new therapeutic strategies., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Insect larval frass has been proposed as a fertilizer and amendment, but methods for testing its effects on plants are poorly developed and need standardization. We obtained different types of black soldier fly ( Hermetia illucens) frass via the factorial combination of (a) two insect diets, as follows: G (Gainesville = 50% wheat bran, 30% alfalfa meal, 20% maize meal) and W (43% sheep whey + 57% seeds); (b) two frass thermal treatments: NT = untreated and T = treated at 70 °C for 1 h. We tested the effects on the germination of cress ( Lepidium sativum L.) and wheat ( Triticum durum Desf.) by applying 1:2 w:w water extracts at 0, 25, 50, 75 and 100% concentration. Standardizing frass water content before extraction affected chemical composition. Frass extracts showed high electrical conductivity (8.88 to 13.78 mS cm -1 ). The W diet was suppressive towards Escherichia coli and showed a lower content of nitrates (e.g., WNT 40% lower than GNT) and a concentration-dependent phytotoxic effect on germinating plants. At 25% concentration, germination indices of G were 4.5 to 40-fold those at 100%. Root and shoot length and root hair area were affected by diet and concentration of frass extracts (e.g., root and shoot length in cress at 25% were, respectively, 4.53 and 2 times higher than at 100%), whereas the effects of the thermal treatment were few or inconclusive. On barley (Hordeum vulgare L.) grown in micropots on a silty loam soil, root mass was reduced by 37% at high extract concentration. A quick procedure for root hair surface area was developed based on the geographic information system (GIS) and may provide a fast method for incorporating root hair phenotyping in frass evaluation. The results indicate that below-ground structures need to be addressed in research on frass effects. For this, phyotoxicity tests should encompass different extract dilutions, and frass water content should be standardized before extraction in the direction of canonical procedures to allow comparisons.

Purpose: Cancer patients with advanced-stage disease have poor prognosis, typically having limited options for efficacious treatment, and genomics-based therapy guidance continues to benefit only a fraction of patients. Next-generation ex vivo approaches, such as cell mass-based response testing (MRT), offer an alternative precision medicine approach for a broader population of patients with cancer, but validation of clinical feasibility and potential impact remain necessary., Materials and Methods: We evaluated the clinical feasibility and accuracy of using live-cell MRT to predict patient drug sensitivity. Using a unified measurement workflow with a 48-hour result turnaround time, samples were subjected to MRT after treatment with a panel of drugs in vitro. After completion of therapeutic course, clinical response data were correlated with MRT-based predictions of outcome. Specimens were collected from 104 patients with solid (n = 69) and hematologic (n = 35) malignancies, using tissue formats including needle biopsies, malignant fluids, bone marrow aspirates, and blood samples. Of the 81 (78%) specimens qualified for MRT, 41 (51%) patients receiving physician-selected therapies had treatments matched to MRT., Results: MRT demonstrated high concordance with clinical responses with an odds ratio (OR) of 14.80 ( P = .0003 [95% CI, 2.83 to 102.9]). This performance held for both solid and hematologic malignances with ORs of 20.67 ( P = .0128 [95% CI, 1.45 to 1,375.57]) and 8.20 ( P = .045 [95% CI, 0.77 to 133.56]), respectively. Overall, these results had a predictive accuracy of 80% ( P = .0026 [95% CI, 65 to 91])., Conclusion: MRT showed highly significant correlation with clinical response to therapy. Routine clinical use is technically feasible and broadly applicable to a wide range of samples and malignancy types, supporting the need for future validation studies.

The chromosome 9p21 locus comprises several tumor suppressor genes including MTAP, CDKN2A, and CDKN2B, and its homo- or heterozygous deletion is associated with reduced survival in multiple cancer types. We report that mice with germ line monoallelic deletion or induced biallelic deletion of the 9p21-syntenic locus (9p21s) developed a fatal myelodysplastic syndrome/myeloproliferative neoplasm (MDS/MPN)-like disease associated with aberrant trabecular bone formation and/or fibrosis in the bone marrow (BM). Reciprocal BM transfers and conditional targeting of 9p21s suggested that the disease originates in the BM stroma. Single-cell analysis of 9p21s-deficient BM stroma revealed the expansion of chondrocyte and osteogenic precursors, reflected in increased osteogenic differentiation in vitro. It also showed reduced expression of factors maintaining hematopoietic stem/progenitor cells, including Cxcl12. Accordingly, 9p21s-deficient mice showed reduced levels of circulating Cxcl12 and concomitant upregulation of the profibrotic chemokine Cxcl13 and the osteogenesis- and fibrosis-related multifunctional glycoprotein osteopontin/Spp1. Our study highlights the potential of mutations in the BM microenvironment to drive MDS/MPN-like disease., (© 2023 by The American Society of Hematology.)

Bone marrow adipose tissue (BMAT) creates a specific microniche within multifunctional bone marrow (BM) ecosystem which imposes changes in surrounding cells and at systemic level. Moreover, BMAT contributes to spatial and temporal separation and metabolic compartmentalization of BM, thus regulating BM homeostasis and diseases. Recent findings have identified novel progenitor subsets of bone marrow adipocytes (BMAd)s recruited during the BM adipogenesis within different skeletal and hematopoietic stem cell niches. Potential of certain mesenchymal BM cells to differentiate into both osteogenic and adipogenic lineages, contributes to the complex interplay of BMAT with endosteal (osteoblastic) niche compartments as an important cellular player in bone tissue homeostasis. Targeting and ablation of BMAT cells at certain states might be an optional and promising strategy for improvement of bone health. Additionally, recent findings demonstrated spatial distribution of BMAds related to hematopoietic cells and pointed out important functional roles in the vital processes such as long-term hematopoiesis. BM adipogenesis appears to be an emergency phenomenon that follows the production of hematopoietic stem and progenitor cell niche factors, thus regulating physiological, stressed, and malignant hematopoiesis. Lipolytic and secretory activity of BMAds can influence survival and proliferation of hematopoietic cells at different maturation stages. Due to their different lipid status, constitutive and regulated BMAds are important determinants of normal and malignant hematopoietic cells. Further elucidation of cellular and molecular players involved in BMAT expansion and crosstalk with malignant cells is of paramount importance for conceiving the new therapies for improvement of BM health., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

In dual-purpose cereal systems, the co-production of fodder and grain can increase farm profitability and reduce farming risks. Our work evaluated shoot and root growth in durum wheat ( Triticum durum Desf.) under dual-purpose management in a medium-high rainfall area of southern Italy. We compared a modern variety (Core) with a tall ancient variety (Saragolle lucana) under traditional (NDP) and dual-purpose (DP) management and tested the hypothesis that clipping plants during the vegetative stage would reduce root growth and dewatering before anthesis, which is advantageous in drought-prone environments. Experiments were conducted in Bella (PZ), Basilicata region, southern Italy (40°42' N, 15°32' E) on a clay loam soil in 2021 in a split-plot design on 2 × 2 main plots and 1 × 2 split-plots with 6 replicates. The DP treatment consisted of simulated grazing by clipping plants at 5 cm from the ground 3 months after sowing (at first hollow stem). Forage Biomass was not different at p = 0.05 between varieties, with an average of 0.58 t ha -1 DM. Grain yield was not penalized by clipping ( p = 0.05) and did not differ significantly between varieties. SPAD was always lower in the Saragolle variety and lowered by clipping. Defoliation delayed phenology in both cultivars but did not reduce the final number of spikes per square meter. Stomatal conductance was correlated to temperature, did not differ between cultivars, and was not influenced by clipping. Soil water depletion was monitored in modern wheat from the booting stage to the beginning of grain filling. Clipping did not result in a reduction in pre-anthesis water depletion, possibly due to evaporative losses. Root density was markedly reduced by clipping in core variety between 0.20 and 0.60 m and much less in Saragolle. Unclipped Saragolle produced thicker roots and higher root masses compared to clipped plants. Defoliated Saragolle shifted to finer roots, reducing root mass more than length. This may have reduced the metabolic cost of soil exploration, thereby increasing root foraging efficiency.

Ancient wheats may be a source of traits that are useful for the tolerance of climate change foreseen conditions of raising temperatures and low water availability. Previous research has shown a fine root system and a high mass of rhizosheath per unit root mass in the italian durum wheat ( Triticum durum Desf) landrace Saragolle Lucana, and this may be relevant for successfully facing adverse conditions during seedling establishment. We investigated the effect of soil water shortage in Saragolle seedlings on root architecture, rhizosheath formation and biomass allocation. Pot experiments were conducted by comparing two levels of soil available water content (AWC): WW (100% of AWC) and DS (50% of AWC). Phenology was delayed by eight days in DS and above and belowground traits were measured at Zadoks 1.3 for each treatment. Biometric data collected at the same phenological stage show that DS plants did not reach the levels of biomass, surface area and space occupation of WW even after attaining the same developmental stage. Namely, plant dimensions were lower at low soil water availability, with the exception of rhizosheath production: DS yielded a 50% increase in rhizosheath mass and 32% increase in rhizosheath mass per unit root mass. The proportion of plant mass reduction in DS was 29.7% for aboveground parts and 34.7% for roots, while reductions in leaf and root surface areas exceeded 43%. The root/shoot mass and area ratios were not significantly different between treatments, and a higher impact on aboveground than on belowground traits at reduced available water was shown only by a lower ratio of shoot height to root depth in DS than in WW. Increases in rhizosheath in absolute and relative terms, which were observed in our experiment in spite of smaller root systems in the ancient durum wheat variety Saragolle lucana at DS, may provide an interesting trait for plant performance in conditions of low soil water availability both for water-related issue and for other effects on plant nutrition and relations with the rhizosphere.

Functional precision medicine offers a promising complement to genomics-based cancer therapy guidance by testing drug efficacy directly on a patient's tumor cells. Here, we describe a workflow that utilizes single-cell mass measurements with inline brightfield imaging and machine-learning based image classification to broaden the clinical utility of such functional testing for cancer. Using these image-curated mass measurements, we characterize mass response signals for 60 different drugs with various mechanisms of action across twelve different cell types, demonstrating an improved ability to detect response for several slow acting drugs as compared with standard cell viability assays. Furthermore, we use this workflow to assess drug responses for various primary tumor specimen formats including blood, bone marrow, fine needle aspirates (FNA), and malignant fluids, all with reports generated within two days and with results consistent with patient clinical responses. The combination of high-resolution measurement, broad drug and malignancy applicability, and rapid return of results offered by this workflow suggests that it is well-suited to performing clinically relevant functional assessment of cancer drug response., (© 2022. The Author(s).)

The Gsα/cAMP signaling pathway mediates the effect of a variety of hormones and factors that regulate the homeostasis of the post-natal skeleton. Hence, the dysregulated activity of Gsα due to gain-of-function mutations (R201C/R201H) results in severe architectural and functional derangements of the entire bone/bone marrow organ. While the consequences of gain-of-function mutations of Gsα have been extensively investigated in osteoblasts and in bone marrow osteoprogenitor cells at various differentiation stages, their effect in adipogenically-committed bone marrow stromal cells has remained unaddressed. We generated a mouse model with expression of Gsα R201C driven by the Adiponectin (Adq) promoter. Adq-Gsα R201C mice developed a complex combination of metaphyseal, diaphyseal and cortical bone changes. In the metaphysis, Gsα R201C caused an early phase of bone resorption followed by bone deposition. Metaphyseal bone formation was sustained by cells that were traced by Adq-Cre and eventually resulted in a high trabecular bone mass phenotype. In the diaphysis, Gsα R201C , in combination with estrogen, triggered the osteogenic activity of Adq-Cre-targeted perivascular bone marrow stromal cells leading to intramedullary bone formation. Finally, consistent with the previously unnoticed presence of Adq-Cre-marked pericytes in intraosseous blood vessels, Gsα R201C caused the development of a lytic phenotype that affected both cortical (increased porosity) and trabecular (tunneling resorption) bone. These results provide the first evidence that the Adq-cell network in the skeleton not only regulates bone resorption but also contributes to bone formation, and that the Gsα/cAMP pathway is a major modulator of both functions., (© 2022. The Author(s).)

The first International Summer School on Bone Marrow Adiposity was organized by members of Bone Marrow Adiposity Society and held virtually on September 6-8 2021. The goal of this meeting was to bring together young scientists interested in learning about bone marrow adipose tissue biology and pathology. Fifty-two researchers from different backgrounds and fields, ranging from bone physiopathology to adipose tissue biology and hematology, participated in the summer school. The meeting featured three keynote lectures on the fundamentals of bone marrow adiposity, three scientific workshops on technical considerations in studying bone marrow adiposity, and six motivational and career development lectures, spanning from scientific writing to academic career progression. Moreover, twenty-one participants presented their work in the form of posters. In this report we highlight key moments and lessons learned from the event., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Labella, Little-Letsinger, Avilkina, Sarkis, Tencerova, Vlug and Palmisano.)

Remodeling of the microenvironment by tumor cells can activate pathways that favor cancer growth. Molecular delineation and targeting of such malignant-cell nonautonomous pathways may help overcome resistance to targeted therapies. Herein we leverage genetic mouse models, patient-derived xenografts, and patient samples to show that acute myeloid leukemia (AML) exploits peripheral serotonin signaling to remodel the endosteal niche to its advantage. AML progression requires the presence of serotonin receptor 1B (HTR1B) in osteoblasts and is driven by AML-secreted kynurenine, which acts as an oncometabolite and HTR1B ligand. AML cells utilize kynurenine to induce a proinflammatory state in osteoblasts that, through the acute-phase protein serum amyloid A (SAA), acts in a positive feedback loop on leukemia cells by increasing expression of IDO1-the rate-limiting enzyme for kynurenine synthesis-thereby enabling AML progression. This leukemia-osteoblast cross-talk, conferred by the kynurenine-HTR1B-SAA-IDO1 axis, could be exploited as a niche-focused therapeutic approach against AML, opening new avenues for cancer treatment., Significance: AML remains recalcitrant to treatments due to the emergence of resistant clones. We show a leukemia-cell nonautonomous progression mechanism that involves activation of a kynurenine-HTR1B-SAA-IDO1 axis between AML cells and osteoblasts. Targeting the niche by interrupting this axis can be pharmacologically harnessed to hamper AML progression and overcome therapy resistance. This article is highlighted in the In This Issue feature, p. 873., (©2022 The Authors; Published by the American Association for Cancer Research.)

Early root traits and allometrics of wheat are important for competition and use of resources. They are under-utilized in research and un-explored in many ancient wheats. This is especially true for the rhizosheath emerging from root-soil interactions. We investigated root morphology, root/shoot relations and the amount of rhizosheath of four tetrapoid wheat seedlings (30 days after emergence): the italian landrace Saragolle Lucana and modern varieties Creso, Simeto and Ciclope, and tested the hypothesis that inoculation with Trichoderma harzianum T-22 (T-22) enhances rhizosheath formation and affects wheat varieties differently. Overall growth of non-inoculated plants showed different patterns in wheat varieties, with Saragolle and Ciclope at the two extremes: Saragolle invests in shoot rather than root mass, and in the occupation of space with highest ( p < 0.05) shoot height to the uppermost internode (5.02 cm) and length-to-mass shoot (97.8 cm g -1 ) and root (more than 140 m g -1 ) ratios. This may be interpreted as maximizing competition for light but also as a compensation for low shoot efficiency due to the lowest ( p < 0.05) recorded values of optically-measured chlorophyll content index (22.8). Ciclope invests in biomass with highest shoot (0.06 g) and root (0.04 g) mass and a thicker root system (average diameter 0.34 mm vs. 0.29 in Saragolle) as well as a highest root/shoot ratio (0.95 g g -1 vs. 0.54 in Saragolle). Rhizosheath mass ranged between 22.14 times that of shoot mass in Ciclope and 43.40 in Saragolle (different for p < 0.05). Inoculation with Trichoderma increased the amount of rhizosheath from 9.4% in Ciclope to 36.1% in Simeto and modified root architecture in this variety more than in others. Ours are the first data on roots and seedling shoot traits of Saragolle Lucana and of Trichoderma inoculation effects on rhizosheath. This opens to new unreported interpretations of effects of Trichoderma inoculation on improving plant growth.

The quality control of probiotic products is the focus of numerous organizations worldwide. Several studies have highlighted the poor microbiological quality of many commercial probiotic formulations in terms of the identity of the contained microorganisms, viability, and purity, thus precluding the expected health benefits and representing a potential health risk for consumers. In this paper, we analyzed the contents of two probiotic formulations, one composed of an encapsulated mixture of lactobacilli and bifidobacteria, and one by a lyophilized yeast. The microorganisms contained in the products were quantified and identified using up-to-date methodologies, such as MALDI-TOF MS and metagenomic analysis. Moreover, as acid and bile tolerance is included among the criteria used to select probiotic microorganisms, in vitro tests were performed to evaluate the behavior of the formulations in conditions mimicking the harsh gastric environment and the intestinal fluids. Our results indicate the high quality of the formulations in terms of the enumeration and identification of the contained organisms, as well as the absence of contaminants. Moreover, both products tolerated the acidic conditions well, with encapsulation providing further protection for the microorganisms. A good tolerance to the simulated artificial intestinal conditions was also evidenced for both preparations.

Blood vessels provide supportive microenvironments for maintaining tissue functions. Age-associated vascular changes and their relation to tissue aging and pathology are poorly understood. Here, we perform 3D imaging of young and aging vascular beds. Multiple organs in mice and humans demonstrate an age-dependent decline in vessel density and pericyte numbers, while highly remodeling tissues such as skin preserve the vasculature. Vascular attrition precedes the appearance of cellular hallmarks of aging such as senescence. Endothelial VEGFR2 loss-of-function mice demonstrate that vascular perturbations are sufficient to stimulate cellular changes coupled with aging. Age-associated tissue-specific molecular changes in the endothelium drive vascular loss and dictate pericyte to fibroblast differentiation. Lineage tracing of perivascular cells with inducible PDGFRβ and NG2 Cre mouse lines demonstrated that increased pericyte to fibroblast differentiation distinguishes injury-induced organ fibrosis and zymosan-induced arthritis. To spur further discoveries, we provide a freely available resource with 3D vascular and tissue maps., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY).)

Age-associated alterations of the hormone-secreting endocrine system cause organ dysfunction and disease states. However, the cell biology of endocrine tissue ageing remains poorly understood. Here, we perform comparative 3D imaging to understand age-related perturbations of the endothelial cell (EC) compartment in endocrine glands. Datasets of a wide range of markers highlight a decline in capillary and artery numbers, but not of perivascular cells in pancreas, testis and thyroid gland, with age in mice and humans. Further, angiogenesis and β-cell expansion in the pancreas are coupled by a distinct age-dependent subset of ECs. While this EC subpopulation supports pancreatic β cells, it declines during ageing concomitant with increased expression of the gap junction protein Gja1. EC-specific ablation of Gja1 restores β-cell expansion in the aged pancreas. These results provide a proof of concept for understanding age-related vascular changes and imply that therapeutic targeting of blood vessels may restore aged endocrine tissue function. This comprehensive data atlas offers over > 1,000 multicolour volumes for exploration and research in endocrinology, ageing, matrix and vascular biology., (© 2020 The Authors. Published under the terms of the CC BY 4.0 license.)

We compared the effects of a nitrogen-containing bisphosphonate (N-BP), zoledronic acid (ZA), and an anti-mouse RANKL antibody (anti-mRANKL Ab) on the bone tissue pathology of a transgenic mouse model of human fibrous dysplasia (FD). For comparison, we also reviewed the histological samples of a child with McCune-Albright syndrome (MAS) treated with Pamidronate for 3 years. EF1α-Gsα R201C mice with FD-like lesions in the tail vertebrae were treated with either 0.2 mg/kg of ZA at day 0, 7, and 14 or with 300 μg/mouse of anti-mRANKL Ab at day 0 and 21. All mice were monitored by Faxitron and histological analysis was performed at day 42. ZA did not affect the progression of the radiographic phenotype in EF1α-Gsα R201C mice. FD-like lesions in the ZA group showed the persistence of osteoclasts, easily detectable osteoclast apoptotic activity and numerous "giant osteoclasts". In contrast, in the anti-mRANKL Ab-treated mice, osteoclasts were markedly reduced/absent, the radiographic phenotype reverted and the FD-like lesions were extensively replaced by newly formed bone. Numerous "giant osteoclasts" were also detected in the samples of the child with MAS. This study supports the hypothesis that osteoclasts per se, independently of their resorptive activity, are essential for development and expansion of FD lesions.

The interest in bone marrow adiposity (BMA) has increased over the last decade due to its association with, and potential role, in a range of diseases (osteoporosis, diabetes, anorexia, cancer) as well as treatments (corticosteroid, radiation, chemotherapy, thiazolidinediones). However, to advance the field of BMA research, standardization of methods is desirable to increase comparability of study outcomes and foster collaboration. Therefore, at the 2017 annual BMA meeting, the International Bone Marrow Adiposity Society (BMAS) founded a working group to evaluate methodologies in BMA research. All BMAS members could volunteer to participate. The working group members, who are all active preclinical or clinical BMA researchers, searched the literature for articles investigating BMA and discussed the results during personal and telephone conferences. According to the consensus opinion, both based on the review of the literature and on expert opinion, we describe existing methodologies and discuss the challenges and future directions for (1) histomorphometry of bone marrow adipocytes, (2) ex vivo BMA imaging, (3) in vivo BMA imaging, (4) cell isolation, culture, differentiation and in vitro modulation of primary bone marrow adipocytes and bone marrow stromal cell precursors, (5) lineage tracing and in vivo BMA modulation, and (6) BMA biobanking. We identify as accepted standards in BMA research: manual histomorphometry and osmium tetroxide 3D contrast-enhanced μCT for ex vivo quantification, specific MRI sequences (WFI and H-MRS) for in vivo studies, and RT-qPCR with a minimal four gene panel or lipid-based assays for in vitro quantification of bone marrow adipogenesis. Emerging techniques are described which may soon come to complement or substitute these gold standards. Known confounding factors and minimal reporting standards are presented, and their use is encouraged to facilitate comparison across studies. In conclusion, specific BMA methodologies have been developed. However, important challenges remain. In particular, we advocate for the harmonization of methodologies, the precise reporting of known confounding factors, and the identification of methods to modulate BMA independently from other tissues. Wider use of existing animal models with impaired BMA production (e.g., Pfrt -/- , Kit W/W-v ) and development of specific BMA deletion models would be highly desirable for this purpose., (Copyright © 2020 Tratwal, Labella, Bravenboer, Kerckhofs, Douni, Scheller, Badr, Karampinos, Beck-Cormier, Palmisano, Poloni, Moreno-Aliaga, Fretz, Rodeheffer, Boroumand, Rosen, Horowitz, van der Eerden, Veldhuis-Vlug and Naveiras.)

Fibrous dysplasia of bone/McCune-Albright syndrome (Polyostotic FD/MAS; OMIM#174800) is a crippling skeletal disease caused by gain-of-function mutations of G s α. Enhanced bone resorption is a recurrent histological feature of FD and a major cause of fragility of affected bones. Previous work suggests that increased bone resorption in FD is driven by RANKL and some studies have shown that the anti-RANKL monoclonal antibody, denosumab, reduces bone turnover and bone pain in FD patients. However, the effect of RANKL inhibition on the histopathology of FD and its impact on the natural history of the disease remain to be assessed. In this study, we treated the EF1α-G s α R201C mice, which develop an FD-like phenotype, with an anti-mouse RANKL monoclonal antibody. We found that the treatment induced marked radiographic and microscopic changes at affected skeletal sites in 2-month-old mice. The involved skeletal segments became sclerotic due to the deposition of new, highly mineralized bone within developing FD lesions and showed a higher mechanical resistance compared to affected segments from untreated transgenic mice. Similar changes were also detected in older mice with a full-blown skeletal phenotype. The administration of anti-mouse RANKL antibody arrested the growth of established lesions and, in young mice, prevented the appearance of new ones. However, after drug withdrawal, the newly formed bone was remodelled into FD tissue and the disease progression resumed in young mice. Taken together, our results show that the anti-RANKL antibody significantly affected the bone pathology and natural history of FD in the mouse. Pending further work on the prevention and management of relapse after treatment discontinuation, our preclinical study suggests that RANKL inhibition may be an effective therapeutic option for FD patients. © 2019 American Society for Bone and Mineral Research., (© 2019 American Society for Bone and Mineral Research.)

Aim: This 4-week, single-centre, randomized, examiner-blind, controlled study investigated the efficacy and safety of 0.15% ethyl lauroyl arginate (LAE)-containing mouthrinse in adults with mild-to-moderate gingivitis., Material and Methods: Subjects were randomized to use 0.15% LAE-containing mouthrinse or 5% hydroalcohol-negative control twice daily after brushing with standard fluoride toothpaste. Plaque, gingivitis and bleeding were assessed at baseline and Weeks 2 and 4. The oral microflora was analysed at baseline and Week 4., Results: Eighty-seven subjects were randomized to treatment. The 0.15% LAE-containing mouthrinse was associated with statistically significantly (p < 0.001) greater reductions in mean plaque and gingivitis scores versus the negative control at Week 2 (difference [95% confidence interval]: plaque 0.83 [0.64, 1.02], 29.1%; gingivitis 0.11 [0.07, 0.14], 4.8%) and Week 4 (co-primary endpoints: plaque 1.23 [1.07, 1.39], 42.6%; gingivitis 0.23 [0.19, 0.28], 10.7%). Bleeding-index scores were significantly (p < 0.001) reduced versus the control at Weeks 2 (by 0.04 [0.03, 0.06], 36.3%) and 4 (by 0.06 [0.04, 0.08], 50.9%). No shifts were detected in the oral microflora. There were no treatment-related adverse events., Conclusions: The 0.15% LAE-containing mouthrinse was well tolerated and significantly reduced plaque, gingivitis and bleeding when used as an adjunct to tooth brushing for 4 weeks., (© 2015 The Authors. Journal of Clinical Periodontology Published by John Wiley & Sons Ltd.)

Objective: To evaluate the effect of desensitising agents on human dentine morphology and composition., Methods: Randomly assigned human coronal-dentine specimens were subjected to: (a) no treatment (smear-layer control, n=4); (b) acid etching with 6% citric acid (demineralised control, n=4); (c) treatment with desensitising agents (12 cycles of 60 s treatment with 60 s between-treatment rinsing, n=6 per agent); and (d) exposure to acidic challenge (pH 5.0 for 90 s, n=6 per agent). The tested products were: Listerine® Advanced Defence Sensitive (LADS; 1.4% potassium oxalate) mouthrinse, Colgate® Sensitive Pro-Relief™ mouthrinse, and toothpaste slurries (paste/water 1:2 wt/wt ratio) of Colgate® Sensitive Pro-Relief™ paste, Crest® Sensitive paste and Sensodyne® Repair and Protect paste. All dentine surfaces were studied by attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, Raman microscopy and high vacuum scanning electron microscopy with energy-dispersive X-ray microanalysis (HV-SEM/EDX)., Results: Desensitising slurry treatments occluded tubule orifices of acid-etched dentine, creating a randomly distributed surface pattern of particle aggregates. The greatest intratubular penetration of occluding particles was found in dentine treated with LADS. The atomic ratios of Ca/N and Ca/P, and the mineral/matrix ratios increased after toothpaste-slurry treatments compared with the acid-etched dentine. However, the acidic challenge removed most surface precipitates and further demineralised these substrates. Before the acidic challenge, the surface features were least affected in specimens treated with Sensodyne® Repair and Protect. After the acidic challenge, the sub-surface occlusion features were least affected in specimens treated with LADS., Clinical Significance: Although most tested products achieved occlusion of dentinal tubules and provided evidence of mineral deposits, the deposit formed by LADS demonstrated the greatest resistance to acidic challenge, which simulates intra-oral demineralisation phases., (Copyright © 2013 Elsevier Ltd. All rights reserved.)