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Metabolism impacts all cellular functions and plays a fundamental role in physiology. Metabolic regulation of hematopoiesis is dynamically regulated under steady-state and stress conditions. It is clear that hematopoietic stem cells (HSCs) impose different energy demands and flexibility during maintenance compared with stressed conditions. However, the cellular and molecular mechanisms underlying metabolic regulation in HSCs remain poorly understood. In this review, we focus on defining the role of fatty acid oxidation (FAO) in HSCs. We first review the existing literature describing FAO in HSCs under steady-state hematopoiesis. Next, we describe the models used to examine HSCs under stress conditions, and, finally, we describe how infection causes a shift toward FAO in HSCs and the impact of using this pathway on emergency hematopoiesis.

Western Ghats are designated as world heritage sites and global biodiversity hotspots. Microbial diversity in this forest area has been largely neglected and is getting attention since the end of the last millennium. In this review, we have studied and organized various microorganisms from Western Ghats and their diversity, important characteristics and potential biotechnological applications. Microorganisms from Western Ghats have been explored individually for potential bioactive molecules. While most of the microorganisms were analyzed for antimicrobial activities, there have been studies on microbial promotion of plant growth. The microbes analyzed included from aquatics, soil, rhizosphere, phyllosphere and even as endophytes. There have been microorganisms which have shown antioxidant and anti-cancerous activities. There are microorganisms capable of degrading plant wastes and also xenobiotics. Microorganisms capable of producing industrially important enzymes have also been reported. The present review explores largely neglected microbial diversity with respect to their ability to produce potential bioactive biomolecules.

A methodology is presented to implement robotic actuators into diaphragm forming of composite laminates. This Hybrid Vacuum-Robotic (HyVR) process is specifically targeted to prevent ‘bridging’ type defects. Examples are presented of successfully forming laminates with deep concave features that would otherwise be impossible by diaphragm forming. The robotically controlled end effectors apply localised pressure to concave regions during the diaphragm forming process. This force application is analogous to the incremental sheet forming process. A generalised methodology is presented which can be applied to develop a bespoke HyVR process for a specific mould. It can then inform what type of end effector should be used and how to apply it within the HyVR process. A key development included in this methodology is the use of the robot to ‘pin’ the laminate to the mould, preventing any unwanted movement. This process could enable automated production of more complex components using diaphragm forming, taking advantage of its lower tooling and equipment costs.

In adult acute myeloid leukemia (AML), the acquisition of driver somatic mutations may be preceded by a benign state termed clonal hematopoiesis (CH). To develop therapeutic strategies to prevent leukemia development from CH, it is important to understand the mechanisms by which CH-driving and AML-driving mutations cooperate. Here, we use mice with inducible mutant alleles common in human CH (DNMT3AR882; mouse Dnmt3aR878H) and AML (NPM1c; mouse Npm1cA). We find that Dnmt3aR878H/+ hematopoietic stem cells (HSCs), but not multipotent progenitor cell (MPP) subsets, have reduced cytokine expression and proinflammatory transcriptional signatures and a functional competitive advantage over their wild-type counterparts. Dnmt3aR878H/+ HSCs are the most potent cell type transformed by Npm1cA, generating myeloid malignancies in which few additional cooperating somatic mutation events were detected. At a molecular level, Npm1cA, in cooperation with Dnmt3aR878H, acutely increased the accessibility of a distinct set of promoters in HSCs compared with MPP cells. These promoters were enriched for cell cycling, PI3K/AKT/mTOR signaling, stem cell signatures, and targets of transcription factors, including NFAT and the chromatin binding factor HMGB1, which have been implicated in human AML. These results demonstrate cooperativity between preexisting Dnmt3aR878H and Npm1cA at the chromatin level, where specific loci altered in accessibility by Npm1cA are dependent on cell context as well as Dnmt3a mutation status. These findings have implications for biological understanding and therapeutic intervention in the transformation from CH to AML.

Clonal hematopoiesis resulting from the enhanced fitness of mutant hematopoietic stem cells (HSC) associates with both favorable and unfavorable health outcomes related to the types of mature mutant blood cells produced, but how this lineage output is regulated is unclear. Using a mouse model of a clonal hematopoiesis–associated mutation, DNMT3AR882/+ (Dnmt3aR878H/+), we found that aging-induced TNFα signaling promoted the selective advantage of mutant HSCs and stimulated the production of mutant B lymphoid cells. The genetic loss of the TNFα receptor TNFR1 ablated the selective advantage of mutant HSCs without altering their lineage output, whereas the loss of TNFR2 resulted in the overproduction of mutant myeloid cells without altering HSC fitness. These results nominate TNFR1 as a target to reduce clonal hematopoiesis and the risk of associated diseases and support a model in which clone size and mature blood lineage production can be independently controlled to modulate favorable and unfavorable clonal hematopoiesis outcomes.Significance:Through the identification and dissection of TNFα signaling as a key driver of murine Dnmt3a-mutant hematopoiesis, we report the discovery that clone size and production of specific mature blood cell types can be independently regulated.See related commentary by Niño and Pietras, p. 2724.This article is highlighted in the In This Issue feature, p. 2711

Supplementary Table from Distinct Tumor Necrosis Factor Alpha Receptors Dictate Stem Cell Fitness versus Lineage Output in Dnmt3a-Mutant Clonal Hematopoiesis

Supplementary Figure from Distinct Tumor Necrosis Factor Alpha Receptors Dictate Stem Cell Fitness versus Lineage Output in Dnmt3a-Mutant Clonal Hematopoiesis

Supplementary methods. Contains sections on generation of rho0 stromal cells, co-culture of primary BMSC and MM cells and extended Seahorse extracellular flux assay methods

Metabolic adjustments are necessary for the initiation, proliferation, and spread of cancer cells. Although mitochondria have been shown to move to cancer cells from their microenvironment, the metabolic consequences of this phenomenon have yet to be fully elucidated. Here, we report that multiple myeloma cells use mitochondrial-based metabolism as well as glycolysis when located within the bone marrow microenvironment. The reliance of multiple myeloma cells on oxidative phosphorylation was caused by intercellular mitochondrial transfer to multiple myeloma cells from neighboring nonmalignant bone marrow stromal cells. This mitochondrial transfer occurred through tumor-derived tunneling nanotubes (TNT). Moreover, shRNA-mediated knockdown of CD38 inhibits mitochondrial transfer and TNT formation in vitro and blocks mitochondrial transfer and improves animal survival in vivo. This study describes a potential treatment strategy to inhibit mitochondrial transfer for clinical benefit and scientifically expands the understanding of the functional effects of mitochondrial transfer on tumor metabolism.Significance:Multiple myeloma relies on both oxidative phosphorylation and glycolysis following acquisition of mitochondria from its bone marrow microenvironment.See related commentary by Boise and Shanmugam, p. 2102

Supplementary figures. Supplementary Fig 1A shows mitochondrial function in BMSC compared to MM cells. Supplementary Fig 1B shows mitochondrial respiration of MM cells cultured with succinate compared to control. Supplementary Fig 2A shows percent of MM cell engraftment in NSG mice. Supplementary Fig 2B shows percent sorted human cells from NSG bone marrow. Supplementary Fig 3A shows quantification of in vitro mitochondrial transfer from BMSC to MM cells. Supplementary Fig 3B shows detection of mtDNA in control and rho0 BMSC. Supplementary Fig 4 shows detection of mitochondrial transfer from BMSC to MM in various culture conditions. Supplementary Fig 5A-C shows quantification of TNT formation between MM cells and BMSC. Supplementary Fig 6 shows TNT anchor points located on BMSC after coculture with MM cell lines. Supplementary Fig 7 shows TNT anchor points located on BMSC after direct coculture with MM cells. Supplementary Fig 8 shows CD38 protein expression in CD38 shRNA targeted cells. Supplementary Fig 9 shows percent engraftment of control KD and CD38 KD MM1S-luc cells in NSG mice.

Supplementary Figure Legend, Methods and Materials from Retinoblastoma Function Is Essential for Establishing Lung Epithelial Quiescence after Injury

Supplementary Figure 1 from Retinoblastoma Function Is Essential for Establishing Lung Epithelial Quiescence after Injury

The retinoblastoma gene product (RB) regulates cell cycle, quiescence, and survival in a cell type–dependent and environment-dependent manner. RB function is critical in the pulmonary epithelium, as evidenced by nearly universal RB inactivation in lung cancer and increased lung cancer risk in persons with germline RB gene mutations. Lung carcinomas occur in the context of epithelial remodeling induced by cytotoxic damage. Whereas the role of RB in development and normal organ homeostasis has been extensively studied, RB function in the context of cellular injury and repair has remained largely unexplored. In the current studies, the RB gene was selectively deleted in the respiratory epithelium of the mouse. Although RB was not required for establishing or maintaining quiescence during lung homeostasis, RB was essential for establishing quiescence during epithelial repair after injury. Notably, aberrant cell cycle progression was sustained for 9 months after injury in RB-deficient lungs. Prenatal and postnatal RB ablation had similar effects, providing evidence that timing of RB loss was not critical to the outcome and that the injury-induced phenotype was not secondary to compensatory alterations occurring during development. These data show that RB is essential for repair of the respiratory epithelium after cytotoxic damage and support a critical unique role for RB in the context of epithelial remodeling after injury. Because human cancers are associated with chronic cellular damage, these findings have important new implications for RB-mediated tumor suppression. [Cancer Res 2008;68(11):4068–76]

With aging, hematopoietic stem cells (HSCs) have an impaired ability to regenerate, differentiate, and produce the entire repertoire of mature blood and immune cells. Due to dysfunctional hematopoiesis, the incidence of hematologic malignancies increases in elderly individuals. Here, we provide an update on HSC-intrinsic and HSC-extrinsic factors and processes recently discovered to contribute to functional decline of HSCs during aging. In addition, we discuss targets and timing of intervention approaches to maintain HSC function during aging and the extent to which these same targets may prevent or delay transformation to hematologic malignancies.

Introduction Trochleoplasty is a surgical procedure used to treat patellar instability by modifying the trochlear groove. Analysis of the groove with a handheld scanner would enable accurate real-time planning and facilitate tailormade correction. We aimed to measure trochlear depth, sulcus angle, trochlear facet ratio, trochlear angle and lateral trochlear inclination angle and to establish inter- and intra-rater reliability for knee models to determine reliability and repeatability. Methods The trochlear grooves of three knee models were scanned by two investigators. Three-dimensional reference models were created and surface-matched. Custom software was used to determine the desired parameters. The intraclass correlation coefficient (ICC) was used to determine test–retest reliability and the parameter results for each model that showed best reproducibility. Results There was good interobserver reliability (trochlear depth, 1.0mm; sulcus angle, 2.7°; trochlear angle, 4.0°; lateral trochlear inclination angle, 4.0°), except in the trochlear facet ratio (32.0%) of one knee model. With outliers removed, the ICC was moderate to excellent in 73.34% of measurements, with trochlear depth showing the best reproducibility. Discussion This feasibility study showed a handheld scanner in conjunction with supporting software can measure trochlear parameters with good to excellent inter- and intra-observer reliability.

The nuances of swimming make the application of traditional return-to-play progressions following concussion challenging. Our purpose was to describe athletic training services and concussion management protocols among National Collegiate Athletic Association swim programs and compare them between the National Collegiate Athletic Association divisions. We surveyed 228 athletic trainers assigned to or with knowledge of their institution’s swim programs from a convenience sample of 539 (response rate = 42.3%) athletic trainers. Athletic training services were provided to 98.6% (214/217) of the programs. Nearly 80% (164/207) of the programs administered baseline concussion testing to swimmers, with differences observed between divisions (p p = .108) in the number of concussions sustained by swimmers in the past academic year were noted between divisions. Specific return-to-swim protocols were reported by 51.8% (115/222) of respondents with no differences in the presence of a specific return-to-swim protocol between divisions (p = .790). While concussions are reported less frequently in swimming than other sports, appropriate medical care, including a specific return-to-swim protocol, is warranted and provided for future use.

Clonal hematopoiesis resulting from the enhanced fitness of mutant hematopoietic stem cells (HSC) associates with both favorable and unfavorable health outcomes related to the types of mature mutant blood cells produced, but how this lineage output is regulated is unclear. Using a mouse model of a clonal hematopoiesis–associated mutation, DNMT3AR882/+ (Dnmt3aR878H/+), we found that aging-induced TNFα signaling promoted the selective advantage of mutant HSCs and stimulated the production of mutant B lymphoid cells. The genetic loss of the TNFα receptor TNFR1 ablated the selective advantage of mutant HSCs without altering their lineage output, whereas the loss of TNFR2 resulted in the overproduction of mutant myeloid cells without altering HSC fitness. These results nominate TNFR1 as a target to reduce clonal hematopoiesis and the risk of associated diseases and support a model in which clone size and mature blood lineage production can be independently controlled to modulate favorable and unfavorable clonal hematopoiesis outcomes. Significance: Through the identification and dissection of TNFα signaling as a key driver of murine Dnmt3a-mutant hematopoiesis, we report the discovery that clone size and production of specific mature blood cell types can be independently regulated. See related commentary by Niño and Pietras, p. 2724. This article is highlighted in the In This Issue feature, p. 2711

Microrobots are motile microsystems constructed using physical, chemical and biological components for operations with respect to definite application. In the present review, we have discussed the various aspects of bio-microrobots, their history and design. While designing a microrobot, two important parameters (and their varities)- actuation and sensing affect the different micromanipulation techniques to be employed (Magnetic, Optical, Electric, fluidic or acoustic). The controlling and actuation system (Vision-based or Force-sensing) selected for the specific application can dictate the fabrication type to be used for manufacture of the microrobot to be used. The controlling parameters are further influenced by the type of propulsion systems, Powering system, and mobility in complex environment and applicability of the microrobot. Presently, the microrobotics has applications in biomedical and environmental engineering. In this review, we have analyzed various aspects of bio-microrobot design, fabrication and applications which can help the future works in nanosciences and bio-based microrobotics.

Background Students with gluten-related disorders (GRD), a spectrum of conditions including celiac disease (CeD) and non-celiac wheat sensitivity (NCWS), often experience challenges when accessing gluten-free (GF) foods. Aims To identify barriers perceived by students with GRD to access GF products on-campus of universities and colleges across Canada. Methods We conducted a cross sectional survey using the RedCap platform and distributed it to the Canadian Celiac Association community. We included students who reported adopting a GFD for various reasons including CeD and other GRD. We collected data on adherence to the GFD using a validated questionnaire (CDAT), presence of perceived barriers to follow a GFD while dining on campus, persistent symptoms, and altered quality of life. Continuous data are expressed as median (IQR), and categorical data as proportions of patients. Mann-Whitney U and Chi2 with Fisher correction were used to assess differences between groups. Results Seventy nine students responded to the survey (5% male and median age = 25 yrs) and 78 had complete data for analysis. Of the 78 students, 52 (66.6%) reported a diagnosis of CeD, while 26 were adopting a GFD for other reasons (non-CeD). The majority were enrolled in university programs (72/78) and 18% were living on-campus. Almost 90% reported difficulties maintaining a GFD while dining on-campus. Similar proportion of CeD and non-CeD reported eating gluten accidentally (75% vs 80%), while 15% reported eating gluten intentionally on-campus at least a few times per week. This was observed more frequently in non-CeD compared with students with CeD (61% vs 17%; p=0.04). Barriers identified in CeD versus non-CeD groups were related to a reduced GF-food variety (48% vs 69%), lack of availability of GF food (21% vs 46%) and increased cost (46% vs 81%) compared with gluten-containing counterparts. The majority of participants were concerned whether the food available on-campus was truly GF (80% vs 54%) as they reported foods not properly labelled. The majority of participants considered their overall health (79%) and quality of life (65%) was fair to terrible while dining on campus. During the pandemic, 76% of them perceived that it was easier to stick to a GF diet. Conclusions Students from various universities and colleges across Canada experience barriers to access GF food on-campus. This has a significant impact on their overall health and quality of life. Proper food labeling, GF certification and improving the variety of GF food on-campus are options for improvement. Funding Agencies None

A simple cost effective and eco-friendly method has been developed to synthesize silver nanoparticles using edible mushroom extract of Oyster mushroom. 0.1Mm silver nitrate (AgNO3) substrate concentration, reducing agent (2.5- and 5.0-ml mushroom extract) and dark incubation (12, 18, 24, 72 hrs.) were assessed for their ability to precise stabilized silver nanoparticles synthesis. Mushroom extract components serve as both reducing and capping agent, while dark incubation acts as a catalyst in the synthesis process. The biosynthesized silver nanoparticles were characterized by dynamic light scattering (DLS) for size, FTIR and UV-Visible spectroscopy. The intensity-based size of biosynthesized silver nanoparticles was 99.30nm which indicated that mushroom extract acted as the reducing and capping agents on the surface of silver nitrate nanoparticles. In vitro antifungal inhibitory activity of biosynthesized silver nanoparticles on colony formation of Rhizoctonia solani were tested by poisoned food technique to observe the inhibitory effect of these green synthesized AgNPs on the mycelium growth of R. solani. Seven different concentrations (150,250, 350,450,550,650 and 750 µl) of mushroom nanoparticles were used. Among, all the concentrations, it is evident that as the concentration of AgNPs increased there was a corresponding increase in growth inhibition of pathogen. The highest growth inhibition (49.73%) was found in the 750 µl which was followed by 650 µl concentrations which recorded 29.59% growth inhibition.