Agricultural nutrients nitrogen and phosphorus can subsequently be transported to waterways and are often managed through the adoption of best management practices (BMPs). However, we have a poor understanding of how the use of BMPs varies spatially and how BMP adoption might be related to nutrient yields in surface waters. To address this, we performed a survey of agricultural landowners across New York State and compared this with estimates of annual incremental nitrogen and phosphorus yields of agricultural origin from the Spatially Referenced Regressions On Watershed attributes (SPARROW) model. Using these socio-behavioral data and SPARROW predictions, we perform colocation analysis to identify areas where watersheds with high nutrient yield from agriculture are collocated with non-use of agricultural BMPs. This colocation analysis offers a novel methodology for identifying areas where monitoring of waterways and promotion of best management practices could be targeted to achieve the greatest benefits.

Cartilage, a flexible and smooth connective tissue that envelops the surfaces of synovial joints, relies on chondrocytes for extracellular matrix (ECM) production and the maintenance of its structural and functional integrity. Melatonin (MT), renowned for its anti-inflammatory and antioxidant properties, holds the potential to modulate cartilage regeneration and degradation. Therefore, the present study was devoted to elucidating the mechanism of MT on chondrocytes. The in vivo experiment consisted of three groups: Sham (only the skin tissue was incised), Model (using the anterior cruciate ligament transection (ACLT) method), and MT (30 mg/kg), with sample extraction following 12 weeks of administration. Pathological alterations in articular cartilage, synovium, and subchondral bone were evaluated using Safranin O-fast green staining. Immunohistochemistry (ICH) analysis was employed to assess the expression of matrix degradation-related markers. The levels of serum cytokines were quantified via Enzyme-linked immunosorbent assay (ELISA) assays. In in vitro experiments, primary chondrocytes were divided into Control, Model, MT, negative control, and inhibitor groups. Western blotting (WB) and Quantitative RT-PCR (q-PCR) were used to detect Silent information regulator transcript-1 (SIRT1)/Nuclear factor kappa-B (NF-κB)/Nuclear factor erythroid-2-related factor 2 (Nrf2)/Transforming growth factor-beta (TGF-β)/Bone morphogenetic proteins (BMPs)-related indicators. Immunofluorescence (IF) analysis was employed to examine the status of type II collagen (COL2A1), SIRT1, phosphorylated NF-κB p65 (p-p65), and phosphorylated mothers against decapentaplegic homolog 2 (p-Smad2). In vivo results revealed that the MT group exhibited a relatively smooth cartilage surface, modest chondrocyte loss, mild synovial hyperplasia, and increased subchondral bone thickness. ICH results showed that MT downregulated the expression of components related to matrix degradation. ELISA results showed that MT reduced serum inflammatory cytokine levels. In vitro experiments confirmed that MT upregulated the expression of SIRT1/Nrf2/TGF-β/BMPs while inhibiting the NF-κB pathway and matrix degradation-related components. The introduction of the SIRT1 inhibitor Selisistat (EX527) reversed the effects of MT. Together, these findings suggest that MT has the potential to ameliorate inflammation, inhibit the release of matrix-degrading enzymes, and improve the cartilage condition. This study provides a new theoretical basis for understanding the role of MT in decelerating cartilage degradation and promoting chondrocyte repair in in vivo and in vitro cultured chondrocytes.

Several differentiation protocols have enabled the generation of intermediate mesoderm (IM)-derived cells from human pluripotent stem cells (hPSC). However, the substantial variability between existing protocols for generating IM cells compromises their efficiency, reproducibility, and overall success, potentially hindering the utility of urogenital system organoids. Here, we examined the role of high levels of Nodal signaling and BMP activity, as well as WNT signaling in the specification of IM cells derived from a UCSD167i-99-1 human induced pluripotent stem cells (hiPSC) line. We demonstrate that precise modulation of WNT and BMP signaling significantly enhances IM differentiation efficiency. Treatment of hPSC with 3 μM CHIR99021 induced TBXT+/MIXL1+ mesoderm progenitor (MP) cells after 48 h of differentiation. Further treatment with a combination of 3 μM CHIR99021 and 4 ng/mL BMP4 resulted in the generation of OSR1+/GATA3+/PAX2+ IM cells within a subsequent 48 h period. Molecular characterization of differentiated cells was confirmed through immunofluorescence staining and RT-qPCR. Hence, this study establishes a consistent and reproducible protocol for differentiating hiPSC into IM cells that faithfully recapitulates the molecular signatures of IM development. This protocol holds promise for improving the success of protocols designed to generate urogenital system organoids in vitro, with potential applications in regenerative medicine, drug discovery, and disease modeling.

X-linked hypophosphatemia (XLH) is caused by mutations in PHEX, leading to rickets and osteomalacia. Adults affected with XLH develop a mineralization of the bone-tendon attachment site (enthesis), called enthesopathy, which causes significant pain and impaired movement. Entheses in mice with XLH (Hyp) have enhanced Bone Morphogenetic Protein (BMP) and Indian hedgehog (IHH) signaling. Treatment of Hyp mice with the BMP signaling blocker palovarotene attenuated BMP/IHH signaling in Hyp entheses, thus indicating that BMP signaling plays a pathogenic role in enthesopathy development and that IHH signaling is activated by BMP signaling in entheses. It was previously shown that mRNA expression of Gdf5 is enhanced in Hyp entheses at P14. Thus, to determine a role for GDF5 in enthesopathy development, Gdf5 was deleted globally in Hyp mice and conditionally in Scx + cells of Hyp mice. In both murine models, BMP/IHH signaling was similarly decreased in Hyp entheses, leading to decreased enthesopathy. BMP/IHH signaling remained unaffected in WT entheses with decreased Gdf5 expression. Moreover, deletion of Gdf5 in Hyp entheses starting at P30, after enthesopathy has developed, partially reversed enthesopathy. Taken together, these results demonstrate that while GDF5 is not essential for modulating BMP/IHH signaling in WT entheses, inappropriate GDF5 activity in Scx + cells contributes to XLH enthesopathy development. As such, inhibition of GDF5 signaling may be beneficial for the treatment of XLH enthesopathy.
X-linked hypophosphatemia (XLH) is a rare bone disorder that leads to short stature and poorly mineralized bones. As adults, patients with XLH often develop a mineralization of the bone-tendon attachment site, called enthesopathy, that results in significant pain. We previously showed that Achilles bone-tendon attachment sites (entheses) in mice with XLH (Hyp) have an enthesopathy characterized by increased Bone Morphogenetic Protein (BMP) signaling. In the current studies, we show that treating Hyp mice with the BMP signaling inhibitor palovarotene prevents enthesopathy, demonstrating that the increased BMP signaling in Hyp entheses leads to enthesopathy development. We also reported that expression of Gdf5, which activates BMP signaling, is enhanced in Hyp entheses. Therefore, to determine if the enhanced Gdf5 expression leads to the increased BMP signaling seen Hyp entheses, Gdf5 was deleted from Hyp mice and also deleted specifically in the entheses of Hyp mice. In both mouse models, enthesopathy development was attenuated, demonstrating that the increased Gdf5 expression in Hyp entheses plays a role in enthesopathy development. These data indicate that blocking GDF5 and BMP signaling may prevent enthesopathy in patients with XLH.

Identifying vulnerable areas to erosion within the watershed and implementing best management practices (BMPs) are crucial steps in mitigating watershed degradation by minimizing sediment yields. The present study evaluates and identifies the BMPs in the Seybouse basin, northeastern Algeria, using the Soil and Water Assessment Tool (SWAT) model. After successful calibration and validation, the model demonstrated a satisfactory ability to simulate monthly discharge and sediment. Then, the calibrated model was employed to evaluate the efficacy of diverse management practices in sediment control. In the SWAT, three soil and conservation practices, as well as vegetated filter strips (VFSs), grade stabilization structures (GSSs), and terracing were evaluated. The average annual sediment yield in the Seybouse watershed is determined to be 14.43 t/ha year, constituting 71% of the total soil loss. VFS demonstrated a sediment reduction of 37.30%, GSS 20.40%, and terracing 42.30%. Among these strategies, terracing results in the greatest reduction, followed by VFS. The results of this study area can be useful for informed decision-making regarding optimal watershed management strategies.

Urban stormwater runoff is a significant source of nutrient pollution that is very costly to treat. Water quality trading (WQT) is a market-based strategy that can be used to lower the costs associated with meeting stormwater quality regulations. While many WQT programs have experienced low participation, Virginia\'s program has seen high participation due to the inclusion of land developers and other regulated stormwater dischargers. However, the extent to which WQT is used as a compliance option by regulated stormwater dischargers is not well understood, particularly when compared with the adoption of traditional compliance options. To address this knowledge gap, we collated a novel dataset comprising site characteristics and stormwater compliance methods for all development projects in the City of Roanoke, Virginia from December 2015 to March 2022. We analyzed this dataset to characterize the adoption of nutrient offset credits and other compliance methods being used, including best management practices (BMPs) and improved land covers associated with reduced nutrient export. Results show that credits are the preferred compliance option in Roanoke and were used as the only treatment compliance method for 59% of projects with treatment requirements. Projects using credits corresponded with a lower median disturbed area (1.36 acres) and lower median nutrient load reduction requirement (0.69 pounds of total phosphorus per year) compared with other compliance methods. Furthermore, we found that 58% of the projects that used credits achieved stormwater quantity compliance using methods other than implementing stormwater control devices. By mapping buyers and sellers of credits, we found that all credit sellers are downstream of the development projects. We discuss how this downstream trading could be a cause for concern, as part of a larger discussion of the advantages of tracking stormwater compliance methods, drawing on Roanoke as a case study.

The detrimental effects of plastic and microplastic accumulation on ecosystems are widely recognized and indisputable. The emergence of biodegradable plastics (BPs) offers a practical solution to plastic pollution. Problematically, however, not all BPs can be fully degraded in the environment. On the contrary, the scientific community has demonstrated that BPs are more likely than conventional plastics (CPs) to degrade into micro/nanoplastics and release additives, which can have similar or even worse effects than microplastics. However, there is very limited information available on the environmental toxicity assessment of BMPs. The absence of a toxicity evaluation system and the uncertainty regarding combined toxicity with other pollutants also impede the environmental toxicity assessment of BMPs. Currently, research is focused on thoroughly exploring the toxic effects of biodegradable microplastics (BMPs). This paper reviews the pollution status of BMPs in the environment, the degradation behavior of BPs and the influencing factors. This paper comprehensively summarizes the ecotoxicological effects of BPs on ecosystems, considering animals, plants, and microorganisms in various environments such as water bodies, soil, and sediment. The focus is on distinguishing between BMPs and conventional microplastics (CMPs). In addition, the combined toxic effects of BMPs and other pollutants are also being investigated. The findings suggest that BMPs may have different or more severe impacts on ecosystems. The rougher and more intricate surface of BMPs increases the likelihood of causing mechanical damage to organisms and breaking down into smaller plastic particles, releasing additives that lead to a series of cascading negative effects on related organisms and ecosystems. In the case of knowledge gaps, future research is also proposed and anticipated to investigate the toxic effects of BMPs and their evaluation.

GREMLIN1 (GREM1) is a secreted protein that antagonizes bone morphogenetic proteins (BMPs). While abnormal GREM1 expression has been reported to cause behavioral defects in postpartum mice, the spatial and cellular distribution of GREM1 in the brain and the influence of the GREM1-secreting cells on brain function and behavior remain unclear. To address this, we designed a genetic cassette incorporating a 3×Flag-TeV-HA-T2A-tdTomato sequence, resulting in the creation of a novel Grem1Tag mouse model, expressing an epitope tag (3×Flag-TeV-HA-T2A) followed by a fluorescent reporter (tdTomato) under the control of the endogenous Grem1 promoter. This design facilitated precise tracking of the cell origin and distribution of GREM1 in the brain using tdTomato and Flag (or HA) markers, respectively. We confirmed that the Grem1Tag mouse exhibited normal motor, cognitive, and social behaviors at postnatal 60 days (P60), compared with C57BL/6J controls. Through immunofluorescence staining, we comprehensively mapped the distribution of GREM1-secreting cells across the central nervous system. Pervasive GREM1 expression was observed in the cerebral cortex (Cx), medulla, pons, and cerebellum, with the highest levels in the Cx region. Notably, within the Cx, GREM1 was predominantly secreted by excitatory neurons, particularly those expressing calcium/calmodulin-dependent protein kinase II alpha (Camk2a), while inhibitory neurons (parvalbumin-positive, PV+) and glial cells (oligodendrocytes, astrocytes, and microglia) showed little or no GREM1 expression. To delineate the functional significance of GREM1-secreting cells, a selective ablation at P42 using a diphtheria toxin A (DTA) system resulted in increased anxiety-like behavior and impaired memory in mice. Altogether, our study harnessing the Grem1Tag mouse model reveals the spatial and cellular localization of GREM1 in the mouse brain, shedding light on the involvement of GREM1-secreting cells in modulating brain function and behavior. Our Grem1Tag mouse serves as a valuable tool for further exploring the precise role of GREM1 in brain development and disease.

Osteoinductive BMPs require a suitable delivery system for treating various pathological conditions of the spine and segmental bone defects. INFUSE, the only commercially available BMP-based osteoinductive device, consisting of rhBMP2 on bovine absorbable collagen sponge (ACS) showed major disadvantages due to serious side effects. A novel osteoinductive device, OSTEOGROW, comprised of rhBMP6 dispersed within autologous blood coagulum (ABC) is a promising therapy for bone regeneration, subjected to several clinical trials for diaphysial bone repair and spinal fusion. In the present study, we have examined the release dynamics showing that the ABC carrier provided a slower, more steady BMP release in comparison to the ACS. Rat subcutaneous assay was employed to evaluate cellular events and the time course of ectopic osteogenesis. The host cellular response to osteoinductive implants was evaluated by flow cytometry, while dynamics of bone formation and maintenance in time were evaluated by histology, immunohistochemistry and micro CT analyses. Flow cytometry revealed that the recruitment of lymphoid cell populations was significantly higher in rhBMP6/ABC implants, while rhBMP2/ACS implants recruited more myeloid populations. Furthermore, rhBMP6/ABC implants more efficiently attracted early and committed progenitor cells. Dynamics of bone formation induced by rhBMP2/ACS was characterized by a delayed endochondral ossification process in comparison to rhBMP6/ABC implants. Besides, rhBMP6/ABC implants induced more ectopic bone volume in all observed time points in comparison to rhBMP2/ACS implants. These results indicate that OSTEOGROW was superior to INFUSE due to ABC\'s advantages as a carrier and rhBMP6 superior efficacy in inducing bone.

UNASSIGNED: To investigate whether the Anti-Müllerian Hormone (AMH), an ovarian hormone belonging to the Transforming Growth Factor β superfamily, may represent a possible candidate for use as a bone anabolic factor.
UNASSIGNED: We performed in vitro studies on Human Osteoblasts (HOb) to evaluate the expression and the functionality of AMHRII, the AMH receptor type-2, and investigate the effects of exogenous AMH exposure on osteogenic gene expression and osteoblast functions.
UNASSIGNED: We reported the first evidence for the expression and functionality of AMHRII in HOb cells, thus suggesting that osteoblasts may represent a specific target for exogenous AMH treatment. Furthermore, the exposure to AMH exerted a stimulatory effect on HOb cells leading to the activation of osteogenic genes, including the upregulation of osteoblastic transcription factors such as RUNX and OSX, along with increased deposition of mineralized nodules.
UNASSIGNED: Our findings proved interesting clues on the stimulatory effects of AMH on mature osteoblasts expressing its specific receptor, AMHRII. This study may therefore have translation value in opening the perspective that AMH may be an effective candidate to counteract the bone loss in osteoporotic patients by selectively targeting osteoblast with minimal off-target effect.