Deactylation by SIRT1 enables liquid-liquid phase separation of IRF3/IRF7 in innate antiviral immunity.

Innate antiviral immunity deteriorates with aging but how this occurs is not entirely clear. Here we identified SIRT1-mediated DNA-binding domain (DBD) deacetylation as a critical step for IRF3/7 activation that is inhibited during aging. Viral-stimulated IRF3 underwent liquid-liquid phase separation (LLPS) with interferon (IFN)-stimulated response element DNA and compartmentalized IRF7 in the nucleus, thereby stimulating type I IFN (IFN-I) expression. SIRT1 deficiency resulted in IRF3/IRF7 hyperacetylation in the DBD, which inhibited LLPS and innate immunity, resulting in increased viral load and mortality in mice. By developing a genetic code expansion orthogonal system, we demonstrated the presence of an acetyl moiety at specific IRF3/IRF7 DBD site/s abolish IRF3/IRF7 LLPS and IFN-I induction. SIRT1 agonists rescued SIRT1 activity in aged mice, restored IFN signaling and thus antagonized viral replication. These findings not only identify a mechanism by which SIRT1 regulates IFN production by affecting IRF3/IRF7 LLPS, but also provide information on the drivers of innate immunosenescence.

Tuning the Mechanical and Geometric Properties of Electrochemically Aligned Collagen Threads Toward Applications in Biohybrid Robotics.

Electrochemically aligned collagen (ELAC) threads fabricated by the isoelectric focusing (IF) of collagen have previously shown potential in tissue engineering and more recently in the fabrication of biohybrid robot structures. For applications in biohybrid robotics, ELAC structures are needed that are both robust and compliant enough to facilitate muscle actuation. However, studies on the effects of IF parameters, and the interactions of such fabrication parameters, on the mechanical and geometric properties of resulting ELAC threads have not been previously found in literature. Understanding the impact of these manufacturing parameters on the material properties is critical to facilitate biohybrid robot design. In this study, the effects of IF duration, IF voltage, and collagen solution concentration were investigated and showed statistically significant effects on adjusting ELAC properties via single-factor experiments. The interactions between parameters exhibited significant joint effects on ELAC property tuning through two-factor experiments. Scanning electron microscopy and 2,4,6-trinitrobenzenesulfonic (TNBS) assays revealed the correlation between high mechanical properties and a combination of low porosity and high degree of crosslinking. By simply tuning IF parameters without changing other fabrication steps, such as crosslinker concentration, ELAC threads with a wide range of mechanical and geometric properties were fabricated. The average tensile modulus of the resulting ELAC threads ranged from 198 ± 90 to 758 ± 138 MPa. The average cross-sectional area ranged from 7756 ± 1000 to 1775 ± 457 µm2. The resultant mapping between IF parameters and ELAC thread properties enabled the production of strong and flexible threads with customizable properties.

High thermal conductivity in soft elastomers with elongated liquid metal inclusions.

Soft dielectric materials typically exhibit poor heat transfer properties due to the dynamics of phonon transport, which constrain thermal conductivity (k) to decrease monotonically with decreasing elastic modulus (E). This thermal-mechanical trade-off is limiting for wearable computing, soft robotics, and other emerging applications that require materials with both high thermal conductivity and low mechanical stiffness. Here, we overcome this constraint with an electrically insulating composite that exhibits an unprecedented combination of metal-like thermal conductivity, an elastic compliance similar to soft biological tissue (Young's modulus < 100 kPa), and the capability to undergo extreme deformations (>600% strain). By incorporating liquid metal (LM) microdroplets into a soft elastomer, we achieve a ∼25× increase in thermal conductivity (4.7 ± 0.2 W⋅m-1⋅K-1) over the base polymer (0.20 ± 0.01 W⋅m-1·K-1) under stress-free conditions and a ∼50× increase (9.8 ± 0.8 W⋅m-1·K-1) when strained. This exceptional combination of thermal and mechanical properties is enabled by a unique thermal-mechanical coupling that exploits the deformability of the LM inclusions to create thermally conductive pathways in situ. Moreover, these materials offer possibilities for passive heat exchange in stretchable electronics and bioinspired robotics, which we demonstrate through the rapid heat dissipation of an elastomer-mounted extreme high-power LED lamp and a swimming soft robot.

A meta-analysis of the effect of CYP2D6 polymorphism on the pharmacokinetics and pharmacodynamics of metoprolol
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OBJECTIVE: To conduct a meta-analysis on the effect of CYP2D6 polymorphism on the pharmacokinetics and pharmacodynamics of metoprolol. METHODS: A systematic review and meta-analysis of studies on the effect of CYP2D6 polymorphism on metoprolol pharmacokinetics and pharmacodynamics was performed by using the China national knowledge infrastructure (CNKI), database for Chinese technical periodicals (VIP), Wanfang, and PubMed databases up to the end of January 2015. Review Manager 5.3 (the coherence collaboration, www.gradepro.org) and comprehensive Meta-Analysis Software v2 (CMA) Biostat, Englewood, NJ, USA) were used for meta-analysis. RESULTS: A total of 567 cases from 7 studies were included in the present study. Meta-analysis results showed that the area under the curve (AUC)0-∞ (RR = -6.75, 95% CI (-9.18, -4.31), p < 0.00001); Cmax (RR = -2.40, 95% CI (-3.25, -1.54), p < 0.00001); T1/2 (RR = -4.81, 95% CI (-6.86, -2.76), p < 0.00001); CL/F (RR = 1.60, 95% CI (1.03,2.17), p < 0.00001); heart rate (RR = 1.48, 95% CI (0.03, 2.92), p = 0.05), systolic blood pressure (RR = -0.69, 95% CI (-1.85,0.47), p = 0.24); and diastolic blood pressure (RR = -1.95, 95% CI (-3.14, -0.76), p = 0.001). Begg's funnel plot test showed that the pharmacokinetic parameters (AUC0-∞, Cmax, T1/2, and CL/F) and pharmacodynamic parameters (HR, DBP, and SBP) were symmetric. Egger's test showed that the pharmacokinetic parameters were asymmetrical, and its intercept was statistically significant (p < 0.05), which was indicative of publication bias. The pharmacodynamic parameter intercept was not statistically significant (p > 0.05), indicating that no publication bias existed. CONCLUSION: CYP2D6 polymorphism significantly influenced the pharmacokinetic parameters of metoprolol. It also affected heart rate and diastolic blood pressure, whereas systolic pressure was not affected.
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The presence of Y chromosome leads to the difference of cancer in patients with different sexes.

(A) In the case of LOY, the proportion of T cells and suppressor macrophages rises and T cells enhance the expression of TOX, eventually leading to the metastasis of the tumor. (B) KRASmut enhances the gene expression of KDM5D, leading to a decreased expression of the AMOT and TAP1/2 downstream of KDM5D. Thereby, the cell-cell junction and antigen presentation are affected. All elements in Figure 1 are original .

Exploring the role of innate immunity in Cholangiocarcinoma: implications for prognosis, immune infiltration, and tumor metastasis.

The innate immune system serves as the body's primary physiological defense against the intrusion of pathogenic microorganisms, playing a pivotal role in restricting viral infections. However, current research on the interplay between innate immune pathways and cancer is limited, with reported effects often inconsistent. Therefore, we aimed to elucidate the relationship between innate immune pathways and tumors through an amalgamation of bioinformatics and extensive data analysis. Conducting a pan-cancer analysis encompassing expression, genomic alterations, and clinical prognosis, we identified a close association between the innate immune pathway and cholangiocarcinoma. Subsequently, our focus shifted to unraveling the role of innate immune pathway proteins in cholangiocarcinoma. TIMER database analysis showed that the innate immune pathway predominantly influences the infiltration of macrophages and B cells in cholangiocarcinoma. Additionally, gene ontology (GO) and pathway analyses were performed for significantly differentially expressed genes correlated with the innate immune pathway in cholangiocarcinoma. Single-cell transcriptome analysis in cholangiocarcinoma demonstrated that genes in the innate immune pathway are primarily expressed in malignant cells, endothelial cells, monocytes and macrophages. To further validate the expression of proteins in the innate immune pathway in the tumor tissues of patients with cholangiocarcinoma, tumor tissue slices from patients with liver intrahepatic cholangiocarcinoma and normal tissue slices from the HPA database were analyzed. These results indicated pronounced activation of the innate immune pathway in the tumor tissues of patients with cholangiocarcinoma. Finally, proteomic data from patients with or without intrahepatic cholangiocarcinoma metastasis were analyzed. The results revealed a significant correlation between the expression and phosphorylation of IKKε and the occurrence of intrahepatic cholangiocarcinoma metastasis. These findings not only demonstrate the significance of the innate immune pathway in cholangiocarcinoma but also its potential as a prospective prognostic biomarker and therapeutic target for this malignancy.

A Preliminary Study on Factors That Drive Patient Variability in Human Subcutaneous Adipose Tissues.

Adipose tissue is a dynamic regulatory organ that has profound effects on the overall health of patients. Unfortunately, inconsistencies in human adipose tissues are extensive and multifactorial, including large variability in cellular sizes, lipid content, inflammation, extracellular matrix components, mechanics, and cytokines secreted. Given the high human variability, and since much of what is known about adipose tissue is from animal models, we sought to establish correlations and patterns between biological, mechanical, and epidemiological properties of human adipose tissues. To do this, twenty-six independent variables were cataloged for twenty patients, which included patient demographics and factors that drive health, obesity, and fibrosis. A factorial analysis for mixed data (FAMD) was used to analyze patterns in the dataset (with BMI > 25), and a correlation matrix was used to identify interactions between quantitative variables. Vascular endothelial growth factor A (VEGFA) and actin alpha 2, smooth muscle (ACTA2) gene expression were the highest loadings in the first two dimensions of the FAMD. The number of adipocytes was also a key driver of patient-related differences, where a decrease in the density of adipocytes was associated with aging. Aging was also correlated with a decrease in overall lipid percentage of subcutaneous tissue, with lipid deposition being favored extracellularly, an increase in transforming growth factor-ß1 (TGFß1), and an increase in M1 macrophage polarization. An important finding was that self-identified race contributed to variance between patients in this study, where Black patients had significantly lower gene expression levels of TGFß1 and ACTA2. This finding supports the urgent need to account for patient ancestry in biomedical research to develop better therapeutic strategies for all patients. Another important finding was that TGFß induced factor homeobox 1 (TGIF1), an understudied signaling molecule, which is highly correlated with leptin signaling, was correlated with metabolic inflammation. Furthermore, this study draws attention to what we define as "extracellular lipid droplets", which were consistently found in collagen-rich regions of the obese adipose tissues evaluated here. Reduced levels of TGIF1 were correlated with higher numbers of extracellular lipid droplets and an inability to suppress fibrotic changes in adipose tissue. Finally, this study indicated that M1 and M2 macrophage markers were correlated with each other and leptin in patients with a BMI > 25. This finding supports growing evidence that macrophage polarization in obesity involves a complex, interconnecting network system rather than a full switch in activation patterns from M2 to M1 with increasing body mass. Overall, this study reinforces key findings in animal studies and identifies important areas for future research, where human and animal studies are divergent. Understanding key drivers of human patient variability is required to unravel the complex metabolic health of unique patients.

Continuous fiber extruder for desktop 3D printers toward long fiber embedded hydrogel 3D printing.

Recent advances in Freeform Reversible Embedding of Suspended Hydrogels (FRESH), a technique that is compatible with most open-source desktop 3D printers, has enabled the fabrication of complex 3D structures using a wide range of natural and synthetic hydrogels, whose mechanical properties can be modified by embedding long fibers into printed hydrogels. However, fiber extruders dedicated for this application are not commercially available or previously reported. To address this, we have designed a continuous fiber extruder (CFE) that is compatible with low-cost, open-source desktop 3D printers, and demonstrated its performance using a Flashforge Creator-pro printer with a Replistruder-2.0 print-head. Key characteristics of the CFE include: (1) it is affordable, accessible and user-friendly to the 3D printing community due to its low fabrication cost and compatibility with open-source hardware and software, (2) it can embed user-defined 2D and 3D features using long fibers into different types of hydrogels, (3) it works with fibers of different mechanical properties and sizes, (4) it can modify mechanical properties of FRESH printed hydrogels via long fiber embedding.

Association Between Neddylation and Immune Response.

Neddylation is a ubiquitin-like post-translational protein modification. It occurs via the activation of the neural precursor cell expressed, developmentally downregulated protein 8 (NEDD8) by three enzymes: activating enzyme, conjugating enzyme, and ligase. NEDD8 was first isolated from the mouse brain in 1992 and was initially considered important for the development and differentiation of the central nervous system. Previously, the downregulation of neddylation was associated with some human diseases, such as neurodegenerative disorders and cancers. In recent years, neddylation has also been proven to be pivotal in various processes of the human immune system, including the regulation of inflammation, bacterial infection, viral infection, and T cell function. Additionally, NEDD8 was found to act on proteins that can affect viral transcription, leading to impaired infectivity. Here, we focused on the influence of neddylation on the innate and adaptive immune responses.

Long-Fiber Embedded Hydrogel 3D Printing for Structural Reinforcement.

Hydrogels are candidate building blocks in a wide range of biomaterial applications including soft and biohybrid robotics, microfluidics, and tissue engineering. Recent advances in embedded 3D printing have broadened the design space accessible with hydrogel additive manufacturing. Specifically, the Freeform Reversible Embedding of Suspended Hydrogels (FRESH) technique has enabled the fabrication of complex 3D structures using extremely soft hydrogels, e.g., alginate and collagen, by assembling hydrogels within a fugitive support bath. However, the low structural rigidity of FRESH printed hydrogels limits their applications, especially those that require operation in nonaqueous environments. In this study, we demonstrated long-fiber embedded hydrogel 3D printing using a multihead printing platform consisting of a custom-built fiber extruder and an open-source FRESH bioprinter with high embedding fidelity. Using this process, fibers were embedded in 3D printed hydrogel components to achieve significant structural reinforcement (e.g., tensile modulus improved from 56.78 ± 8.76 to 382.55 ± 25.29 kPa and tensile strength improved from 9.44 ± 2.28 to 45.05 ± 5.53 kPa). In addition, we demonstrated the versatility of this technique by using fibers of a wide range of sizes and material types and implementing different 2D and 3D embedding patterns, such as embedding a conical helix using electrochemically aligned collagen fiber via nonplanar printing. Moreover, the technique was implemented using low-cost material and is compatible with open-source software and hardware, which facilitates its adoption and modification for new research applications.

LATS1 is a central signal transmitter for achieving full type-I interferon activity.

Interferons (IFNs) have broad-spectrum antiviral activity to resist virus epidemic. However, IFN antiviral efficacy needs to be greatly improved. Here, we reveal that LATS1 is a vital signal transmitter governing full type-I IFN (IFN-I) signaling activity. LATS1 constitutively binds with the IFN-I receptor IFNAR2 and is rapidly tyro-phosphorylated by Tyk2 upon IFN-I engagement. Tyro-phosphorylation of LATS1 promotes LATS1 activation and YAP degradation, thereby promoting IFN-mediated antiproliferation activity. Moreover, activated LATS1 translocates into the nucleus and induces CDK8-Ser62 phosphorylation, which in turn phosphorylates STAT1 at Ser727 and induces full IFN-I antiviral activity. LATS1 deficiency restricts in vivo IFN-I signaling and attenuates host antiviral immune response. Our study identifies IFN-I as a previously unidentified extracellular diffusible ligand signal for activation of the Hippo core LATS1 pathway and reveals Tyk2-LATS1-CDK8 as a complete signaling cascade controlling full IFN-I activity.

Function of Protein S-Palmitoylation in Immunity and Immune-Related Diseases.

Protein S-palmitoylation is a covalent and reversible lipid modification that specifically targets cysteine residues within many eukaryotic proteins. In mammalian cells, the ubiquitous palmitoyltransferases (PATs) and serine hydrolases, including acyl protein thioesterases (APTs), catalyze the addition and removal of palmitate, respectively. The attachment of palmitoyl groups alters the membrane affinity of the substrate protein changing its subcellular localization, stability, and protein-protein interactions. Forty years of research has led to the understanding of the role of protein palmitoylation in significantly regulating protein function in a variety of biological processes. Recent global profiling of immune cells has identified a large body of S-palmitoylated immunity-associated proteins. Localization of many immune molecules to the cellular membrane is required for the proper activation of innate and adaptive immune signaling. Emerging evidence has unveiled the crucial roles that palmitoylation plays to immune function, especially in partitioning immune signaling proteins to the membrane as well as to lipid rafts. More importantly, aberrant PAT activity and fluctuations in palmitoylation levels are strongly correlated with human immunologic diseases, such as sensory incompetence or over-response to pathogens. Therefore, targeting palmitoylation is a novel therapeutic approach for treating human immunologic diseases. In this review, we discuss the role that palmitoylation plays in both immunity and immunologic diseases as well as the significant potential of targeting palmitoylation in disease treatment.

3D Printing Hydrogel-Based Soft and Biohybrid Actuators: A Mini-Review on Fabrication Techniques, Applications, and Challenges.

Stimuli-responsive hydrogels are candidate building blocks for soft robotic applications due to many of their unique properties, including tunable mechanical properties and biocompatibility. Over the past decade, there has been significant progress in developing soft and biohybrid actuators using naturally occurring and synthetic hydrogels to address the increasing demands for machines capable of interacting with fragile biological systems. Recent advancements in three-dimensional (3D) printing technology, either as a standalone manufacturing process or integrated with traditional fabrication techniques, have enabled the development of hydrogel-based actuators with on-demand geometry and actuation modalities. This mini-review surveys existing research efforts to inspire the development of novel fabrication techniques using hydrogel building blocks and identify potential future directions. In this article, existing 3D fabrication techniques for hydrogel actuators are first examined. Next, existing actuation mechanisms, including pneumatic, hydraulic, ionic, dehydration-rehydration, and cell-powered actuation, are reviewed with their benefits and limitations discussed. Subsequently, the applications of hydrogel-based actuators, including compliant handling of fragile items, micro-swimmers, wearable devices, and origami structures, are described. Finally, challenges in fabricating functional actuators using existing techniques are discussed.

Protein N-myristoylation: functions and mechanisms in control of innate immunity.

Protein N-myristoylation is an important fatty acylation catalyzed by N-myristoyltransferases (NMTs), which are ubiquitous enzymes in eukaryotes. Specifically, attachment of a myristoyl group is vital for proteins participating in various biological functions, including signal transduction, cellular localization, and oncogenesis. Recent studies have revealed unexpected mechanisms indicating that protein N-myristoylation is involved in host defense against microbial and viral infections. In this review, we describe the current understanding of protein N-myristoylation (mainly focusing on myristoyl switches) and summarize its crucial roles in regulating innate immune responses, including TLR4-dependent inflammatory responses and demyristoylation-induced innate immunosuppression during Shigella flexneri infection. Furthermore, we examine the role of myristoylation in viral assembly, intracellular host interactions, and viral spread during human immunodeficiency virus-1 (HIV-1) infection. Deeper insight into the relationship between protein N-myristoylation and innate immunity might enable us to clarify the pathogenesis of certain infectious diseases and better harness protein N-myristoylation for new therapeutics.

A Dual Role of Type I Interferons in Antitumor Immunity.

Type I interferons (IFN-Is) are a family of cytokines that exert direct antiviral effects and regulate innate and adaptive immune responses through direct and indirect mechanisms. It is generally believed that IFN-Is repress tumor development via restricting tumor proliferation and inducing antitumor immune responses. However, recent emerging evidence suggests that IFN-Is play a dual role in antitumor immunity. That is, in the early stage of tumorigenesis, IFN-Is promote the antitumor immune response by enhancing antigen presentation in antigen-presenting cells and activating CD8+ T cells. However, in the late stage of tumor progression, persistent expression of IFN-Is induces the expression of immunosuppressive factors (PD-L1, IDO, and IL-10) on the surface of dendritic cells and other bone marrow cells and inhibits their antitumor immunity. This review outlines these dual functions of IFN-Is in antitumor immunity and elucidates the involved mechanisms, as well as their applications in tumor therapy.

Measuring the transverse Young's modulus of maize rind and pith tissues.

Wind-induced bending loads frequently cause failure of maize (corn) stalks. When failure occurs, it usually manifests as transverse buckling. Because this failure mode is closely related to transverse tissue stiffness, the purpose of this study was to develop a method for measuring the transverse Young's modulus of maize stalk rind and pith tissues. Short, disc-shaped stalk segments were used for this purpose. X-ray computed tomography was used to obtain the geometry of each specimen prior to testing. Each specimen was tested in two different configurations. Computed tomography data was used to create a specimen-specific finite element model of each test specimen. Data from the first testing configuration was used in conjunction with the finite element model to determine the Young's Modulus values for each specimen. The specimen-specific finite element models provided estimates of the stress states in the stem under transverse loading, and these stress states accurately predicted the location of failure in transverse test specimens. The entire testing method was validated using data from one test configuration to predict the structural response of each specimen during the second test configuration.

DARLING: a device for assessing resistance to lodging in grain crops.

BACKGROUND: Stalk lodging (breakage of plant stems prior to harvest) is a major problem for both farmers and plant breeders. A limiting factor in addressing this problem is the lack of a reliable method for phenotyping stalk strength. Previous methods of phenotyping stalk strength induce failure patterns different from those observed in natural lodging events. This paper describes a new device for field-based phenotyping of stalk strength called "DARLING" (device for assessing resistance to lodging in grains). The DARLING apparatus consists of a vertical arm which is connected to a horizontal footplate by a hinge. The operator places the device next to a stalk, aligns the stalk with a force sensor, steps on the footplate, and then pushes the vertical arm forward until the stalk breaks. Force and rotation are continuously recorded during the test and these quantities are used to calculate two quantities: stalk flexural stiffness and stalk bending strength. RESULTS: Field testing of DARLING was performed at multiple sites. Validation was based upon several factors. First, the device induces the characteristic "crease" or Brazier buckling failure patterns observed in naturally lodged stalks. Second, in agreement with prior research, flexural rigidity values attained using the DARLING apparatus are strongly correlated with bending strength measurements. Third, flexural stiffness and bending strength values obtained with DARLING are in agreement with laboratory-based stiffness and strength values for corn stalks. Finally, a paired specimen experimental design was used to determine that the flexural data obtained with DARLING is in agreement with laboratory-based flexural testing results of the same specimens. DARLING was also deployed in the field to assess phenotyping throughput (# of stalks phenotyped per hour). Over approximately 5000 tests, the average testing rate was found to be 210 stalks/h. CONCLUSIONS: The DARLING apparatus provides a quantitative assessment of stalk strength in a field setting. It induces the same failure patterns observed in natural lodging events. DARLING can also be used to perform non-destructive flexural tests. This technology has many applications, including breeding, genetic studies on stalk strength, longitudinal studies of stalk flexural stiffness, and risk assessment of lodging propensity.

The elastic modulus for maize stems.

BACKGROUND: Stalk lodging is a serious challenge in the production of maize and sorghum. A comprehensive understanding of lodging will likely require accurate characterizations of the mechanical properties of such plants. One of the most important mechanical properties for structural analysis of bending is the modulus of elasticity. The purpose of this study was to measure the modulus of elasticity of dry, mature maize rind tissues using three different loading modes (bending, compression and tensile), and to determine the accuracy and reliability of each test method. RESULTS: The three testing modes produced comparable elastic modulus values. For the sample in this study, modulus values ranged between 6 and 16 GPa. All three testing modes exhibited relatively favorable repeatability (i.e. test-to-test variation of < 5%). Modulus values of internodal specimens were significantly higher than specimens consisting of both nodal and internodal tissues, indicating spatial variation in the modulus of elasticity between the nodal and internodal regions. CONCLUSIONS: Bending tests were found to be the least labor intensive method and also demonstrated the best test-to-test repeatability. This test provides a single aggregate stiffness value for an entire stalk. Compression tests were able to determine more localized (i.e., spatially dependent) modulus of elasticity values, but required additional sample preparation and test time. Finally, tensile tests provided the most focused measurements of the modulus of elasticity, but required the longest sample preparation time.

Measuring the compressive modulus of elasticity of pith-filled plant stems.

BACKGROUND: The compressional modulus of elasticity is an important mechanical property for understanding stalk lodging, but this property is rarely available for thin-walled plant stems such as maize and sorghum because excised tissue samples from these plants are highly susceptible to buckling. The purpose of this study was to develop a testing protocol that provides accurate and reliable measurements of the compressive modulus of elasticity of the rind of pith-filled plant stems. The general approach was to relying upon standard methods and practices as much as possible, while developing new techniques as necessary. RESULTS: Two methods were developed for measuring the compressional modulus of elasticity of pith-filled node-node specimens. Both methods had an average repeatability of ± 4%. The use of natural plant morphology and architecture was used to avoid buckling failure. Both methods relied up on spherical compression platens to accommodate inaccuracies in sample preparation. The effect of sample position within the test fixture was quantified to ensure that sample placement did not introduce systematic errors. CONCLUSIONS: Reliable measurements of the compressive modulus of elasticity of pith-filled plant stems can be performed using the testing protocols presented in this study. Recommendations for future studies were also provided.