“Take My Bone Away?” Hypoxia and bone: A narrative review

To maintain normal cellular and physiological function, sufficient oxygen is required. Recently, evidence has suggested that hypoxia, either pathological or environmental, may influence bone health. It appears that bone cells are distinctly responsive to hypoxic stimuli; for better or worse, this is still yet to be elucidated. Hypoxia has been shown to offer potentially therapeutic effects for bone by inducing an osteogenic–angiogenic response, although, others have noted excessive osteoclastic bone resorption instead. Much evidence suggests that the hypoxic‐inducible pathway is integral in mediating the changes in bone metabolism. Furthermore, many factors associated with hypoxia including changes in energy metabolism, acid–base balance and the increased generation of reactive oxygen species, are known to influence bone metabolism. This review aims to examine some of the putative mechanisms responsible for hypoxic‐induced alterations of bone metabolism, with regard to osteoclasts and osteoblasts, both positive and negative.     

Lesser Prairie-Chicken Survival in Varying Densities of Energy Development

Anthropogenic features increasingly affect ecological processes with increasing human demand for natural resources. Such effects also have the potential to vary depending on the sex and age of an individual because of inherent behavioral and life experience differences. For the lesser prairie-chicken (Tympanuchus pallidicinctus), studies on male survival are limited because most previous research has been focused on females. To better understand patterns of lesser prairie-chicken survival in habitat with varying levels of anthropogenic infrastructure associated with oil and natural gas development, we monitored survival of 178 radio-tagged male and female lesser prairie-chickens in eastern New Mexico, USA, from 2013 to 2015. We examined the relationships of shrub cover, proximity to and density of anthropogenic features (i.e., utility poles), displacement of natural vegetation by anthropogenic features (i.e., area of roads and well pads), and individual demographics (i.e., sex, age) with lesser prairie-chicken survival. Furthermore, we categorized the probable cause of mortality and examined its relationship with oil and gas development intensity (indexed by utility pole density) within 1,425 m of an individual's mortality site or final observed location. We predicted that survival would be lower for individuals exposed to greater levels of anthropogenic features, and that males and subadults would be more negatively affected than females and adults because of increased exposure to predators during the lekking season and naiveté. Relationships between survival and utility pole density, sex, and age were supported in our top-ranked models, whereas models including other anthropogenic and natural features (i.e., roads, well pads, shrub cover) received little support. We predicted a substantial decrease in adult and subadult male survival with increasing densities of utility poles. The relationship between survival and utility pole density for females was weaker and not as clearly supported as for males. We did not find a detectable difference in utility pole counts among probable mortality causes. Our findings highlight the importance of including male lesser prairie-chickens in research and conservation planning, and the negative effect that high densities of anthropogenic features can have on lesser prairie-chicken survival. © 2021 The Wildlife Society.     

Spatially Explicit Assessment of Sandhill Crane Exposure to Potential Transmission Line Collision Risk

Infrastructure development can affect avian populations through direct collision mortality. Estimating the exposure of local bird populations to the risk of direct mortality from infrastructure development requires site- and species-specific data, which managers may find difficult to obtain at the scale over which management decisions are made. We quantify the potential exposure of sandhill cranes (Antigone canadensis) to collision with horizontal structures (e.g., transmission lines) within vital wintering grounds of the Middle Rio Grande Valley (MRGV), New Mexico, USA, 2014–2020. Limited maneuverability and visual acuity make sandhill cranes vulnerable to collisions with infrastructure bisecting their flight paths. We used data from 81 global positioning system (GPS)-tagged cranes to estimate the spatially explicit flight height distribution along the MRGV, the passage rate across hypothetical transmission lines, and the resulting exposure rate (exposed passes/crane/day). The exposure rate ranged from 0–0.28 exposed passes/crane/day (median = 0.015) assuming an exposure zone of 7–60 m above ground level, and identified hotspots of potential exposure within the MRGV. Mapped exposure rates can assist in the siting of proposed high-voltage transmission lines, or other infrastructure, to limit effects on sandhill cranes and other avian species at risk of collision. Our approach can be replicated and applied in similar situations where birds are exposed to possible collision with power lines. © 2021 The Authors. The Journal of Wildlife Management published by Wiley Periodicals LLC on behalf of The Wildlife Society.     

Role of HRTPT in kidney proximal epithelial cell regeneration: Integrative differential expression and pathway analyses using microarray and scRNA-seq

Damage to proximal tubules due to exposure to toxicants can lead to conditions such as acute kidney injury (AKI), chronic kidney disease (CKD) and ultimately end-stage renal failure (ESRF). Studies have shown that kidney proximal epithelial cells can regenerate particularly after acute injury. In the previous study, we utilized an immortalized in vitro model of human renal proximal tubule epithelial cells, RPTEC/TERT1, to isolate HRTPT cell line that co-expresses stem cell markers CD133 and CD24, and HREC24T cell line that expresses only CD24. HRTPT cells showed most of the key characteristics of stem/progenitor cells; however, HREC24T cells did not show any of these characteristics. The goal of this study was to further characterize and understand the global gene expression differences, upregulated pathways and gene interaction using scRNA-seq in HRTPT cells. Affymetrix microarray analysis identified common gene sets and pathways specific to HRTPT and HREC24T cells analysed using DAVID, Reactome and Ingenuity software. Gene sets of HRTPT cells, in comparison with publicly available data set for CD133+ infant kidney, urine-derived renal progenitor cells and human kidney-derived epithelial proximal tubule cells showed substantial similarity in organization and interactions of the apical membrane. Single-cell analysis of HRTPT cells identified unique gene clusters associated with CD133 and the 92 common gene sets from three data sets. In conclusion, the gene expression analysis identified a unique gene set for HRTPT cells and narrowed the co-expressed gene set compared with other human renal–derived cell lines expressing CD133, which may provide deeper understanding in their role as progenitor/stem cells that participate in renal repair.     

Critical considerations for the development of potency tests for therapeutic applications of mesenchymal stromal cell-derived small extracellular vesicles

Mesenchymal stromal/stem cells (MSCs) have been widely tested against many diseases, with more than 1000 registered clinical trials worldwide. Despite many setbacks, MSCs have been approved for the treatment of graft-versus-host disease and Crohn disease. However, it is increasingly clear that MSCs exert their therapeutic functions in a paracrine manner through the secretion of small extracellular vesicles (sEVs) of 50–200 nm in diameter. Unlike living cells that can persist long-term, sEVs are non-living and non-replicative and have a transient presence in the body. Their small size also renders sEV preparations highly amenable to sterilization by filtration. Together, acellular MSC-sEV preparations are potentially safer and easier to translate into the clinic than cellular MSC products. Nevertheless, there are inherent challenges in the development of MSC-sEV drug products. MSC-sEVs are products of living cells, and living cells are sensitive to changes in the external microenvironment. Consequently, quality control metrics to measure key identity and potency features of MSC-sEV preparations have to be specified during development of MSC-sEV therapeutics. The authors have previously described quantifiable assays to define the identity of MSC-sEVs. Here the authors discuss requirements for prospective potency assays to predict the therapeutic effectiveness of the drug substance in accordance with International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use guidelines. Although potency assays should ideally reflect the mechanism of action (MoA), this is challenging because the MoA for the reported efficacy of MSC-sEV preparations against multiple diseases of diverse underlying pathology is likely to be complex and different for each disease and difficult to fully elucidate. Nevertheless, robust potency assays could be developed by identifying the EV attribute most relevant to the intended biological activity in EV-mediated therapy and quantifying the EV attribute. Specifically, the authors highlight challenges and mitigation measures to enhance the manufacture of consistent and reproducibly potent sEV preparations, to identify and select the appropriate EV attribute for potency assays despite a complex “work-in-progress” MoA and to develop assays likely to be compliant with regulatory guidance for assay validation.     

MAPK signaling regulates c‐MYC for melanoma cell adaptation to asparagine restriction

Amino acid restriction is among promising potential cancer treatment strategies. However, cancer cells employ a multitude of mechanisms to mount resistance to amino acid restriction, which impede the latter’s clinical development. Here we show that MAPK signaling activation in asparagine‐restricted melanoma cells impairs GSK3‐β‐mediated c‐MYC degradation. In turn, elevated c‐MYC supports ATF4 translational induction by enhancing the expression of the amino acid transporter SLC7A5, increasing the uptake of essential amino acids, and the subsequent maintenance of mTORC1 activity in asparagine‐restricted melanoma cells. Blocking the MAPK‐c‐MYC‐SLC7A5 signaling axis cooperates with asparagine restriction to effectively suppress melanoma cell proliferation. This work reveals a previously unknown axis of cancer cell adaptation to asparagine restriction and informs mechanisms that may be targeted for enhanced therapeutic efficacy of asparagine limiting strategies.     

Effect of Non-Volatile Constituents of Elsholtzia ciliata (Thunb.) Hyl. from Southern Vietnam on Reactive Oxygen Species and Nitric Oxide Release in Macrophages

The extract of Elsholtzia ciliata aerial parts was subjected to bio-guided isolation using the intercellular ROS reduction in J774A.1 macrophages to monitor the anti-oxidative activity. Fifteen compounds were isolated from the active fractions including eleven flavonoids (vitexin, pedalin, luteolin-7-O-β-d -glucopyranoside, apigenin-5-O-β-d -glucopyranoside, apigenin-7-O-β-d -glucopyranoside, chrysoeriol-7-O-β-d -glucopyranoside, 7,3′-dimethoxyluteolin-6-O-β-d -glucopyranoside, luteolin, 5,6,4′-trihydroxy-7,3′-dimethoxyflavone, 5-hydroxy-6,7-dimethoxyflavone (compound 13 ), 5-hydroxy-7,8-dimethoxyflavone); three hydroxycinnamic acid derivatives (caffeic acid, 4-(E)-caffeoyl-l -threonic acid, 4-O-(E)-p-coumaroyl-l -threonic acid) and one fatty acid (α-linolenic acid). The biological evaluation of these compounds (10–2.5 μm ) indicated that all of them exerted good antioxidant and anti-inflammatory activities, in particular compound 13 .