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The Eagle Bioscience’s Noradrenaline (Norepinephrine) Sensitive ELISA was utilized in two recent publications! On study explored how norepinephrine transporter defects lead to sympathetic hyperactivity in familial dysautonomia models and the other focused on the effects of a caffeine containing pre-workout supplement on β₂-adrenergic and MAPK signaling during resistance exercise. Check out the full text and abstracts below.

Norepinephrine transporter defects lead to sympathetic hyperactivity in Familial Dysautonomia models

Abstract

Familial dysautonomia (FD), a rare neurodevelopmental and neurodegenerative disorder affects the sympathetic and sensory nervous system. Although almost all patients harbor a mutation in ELP1, it remains unresolved exactly how function of sympathetic neurons (symNs) is affected; knowledge critical for understanding debilitating disease hallmarks, including cardiovascular instability or dysautonomic crises, that result from dysregulated sympathetic activity. Here, we employ the human pluripotent stem cell (hPSC) system to understand symN disease mechanisms and test candidate drugs. FD symNs are intrinsically hyperactive in vitro, in cardiomyocyte co-cultures, and in animal models. We report reduced norepinephrine transporter expression, decreased intracellular norepinephrine (NE), decreased NE re-uptake, and excessive extracellular NE in FD symNs. SymN hyperactivity is not a direct ELP1 mutation result, but may connect to NET via RAB proteins. We found that candidate drugs lowered hyperactivity independent of ELP1 modulation. Our findings may have implications for other symN disorders and may allow future drug testing and discovery.

Wu, HF., Yu, W., Saito-Diaz, K. et al. Norepinephrine transporter defects lead to sympathetic hyperactivity in Familial Dysautonomia models. Nat Commun 13, 7032 (2022). https://doi.org/10.1038/s41467-022-34811-7

The effects of a caffeine containing pre-workout supplement on β2-adrenergic and MAPK signaling during resistance exercise

Abstract

Aim: The acute myocellular responses of caffeine supplementation during resistance exercise (RE) have not been investigated. β₂-Adrenergic receptors (β₂AR) may be a target of the stimulatory effects of caffeine and stimulate bioenergetic pathways including protein kinase A (PKA), and mitogen-activated protein kinases (MAPK).
Purpose: Elucidate the effects of pre-workout supplementation on signaling responses to an acute RE bout.
Methods: In a randomized, counter-balanced, double-blind, placebo-controlled, within-subject crossover study, ten resistance-trained males (mean ± SD; age = 22 ± 2.4 years, height = 175 ± 7 cm, body mass = 84.1 ± 11.8 kg) consumed a caffeine containing multi-ingredient pre-workout supplement (SUPP) or color and flavor matched placebo (PL) 60 min prior to an acute RE bout of barbell back squats. Pre- and post-exercise muscle biopsies were analyzed for the phosphorylation (p-) of β₂AR, PKA, and MAPK (ERK, JNK, p38). Epinephrine was determined prior to supplementation (baseline; BL), after supplementation but prior to RE (PRE), and immediately after RE (POST).
Results: Epinephrine increased at PRE in SUPP (mean ± SE: 323 ± 34 vs 457 ± 68 pmol/l; p = 0.028), and was greatest at POST in the SUPP condition compared to PL (5140 ± 852 vs 2862 ± 498 pmol/l; p = 0.006). p-β₂AR and p-MAPK increased post-exercise (p < 0.05) with no differences between conditions (p > 0.05). Pearson correlations indicated there was a relationship between epinephrine and p-β₂AR in PL (r = − 0.810; p = 0.008), and p-β₂AR and ERK in SUPP (r = 0.941; p < 0.001).
Conclusion: Consumption of a caffeine containing pre-workout supplement improves performance, possibly through increases in pre-exercise catecholamines. However, the acute myocellular signaling responses were largely similar post-exercise.

Nicoll, J.X., Fry, A.C. & Mosier, E.M. The effects of a caffeine containing pre-workout supplement on β2-adrenergic and MAPK signaling during resistance exercise. Eur J Appl Physiol 123, 585–599 (2023). https://doi.org/10.1007/s00421-022-05085-0

If you have any questions about our Noradrenaline (Norepinephrine) Sensitive ELISA or our other offerings, please contact us here.

The Eagle Bioscience’s High Sensitive CRP ELISA was utilized in a recent publication exploring risk factors for cardiometabolic disease in professional firefighters. Check out the full text and abstract below.

Abstract

Objective: Firefighters are plagued with cardiometabolic disease (CMD). Obesity, poor cardiorespiratory and muscular fitness, and blood lipids (LDL-C, triglycerides, low HDL-C) are risk factors for CMD. However, markers of oxidative stress, inflammation and insulin resistance can provide further insight regarding CMD risk.

Methods: This study investigated the relationships between fitness metrics (cardiorespiratory and muscular fitness, percent body fat, waist circumference), blood lipids, blood pressure, and years of experience as a firefighter to blood markers of insulin resistance: (homeostatic model assessment for insulin resistance, HOMA-IR), oxidative stress: advanced oxidation protein products (AOPP) and inflammation: C-reactive protein (CRP).

Results: Waist circumference and blood concentrations of triglycerides were significantly related to AOPP and HOMA-IR. Cardiorespiratory fitness was inversely related to AOPP, HOMA-IR and CRP.

Conclusion: These findings demonstrate the importance of high cardiorespiratory fitness and low waist circumference to reduce markers of CMD.

McAllister, Matthew J., et al. “Risk Factors for Cardiometabolic Disease in Professional Firefighters.” Journal of Occupational & Environmental Medicine, Publish Ahead of Print, 2022, https://doi.org/10.1097/jom.0000000000002743.

If you have any questions about the High Sensitive CRP ELISA or our other offerings, please contact us here.

The Eagle Bioscience’s Ultra-Sensitive Streptavidin Coated Clear 96 Well Microplate was utilized in a recent publication that explored non-viral precision T cell receptor replacement for personalized cell therapy. Check out the full text and abstract below.

Abstract

T cell receptors (TCRs) enable T cells to specifically recognize mutations in cancer cells. Here we developed a clinical-grade approach based on CRISPR–Cas9 non-viral precision genome-editing to simultaneously knockout the two endogenous TCR genes TRAC (which encodes TCRα) and TRBC (which encodes TCRβ). We also inserted into the TRAC locus two chains of a neoantigen-specific TCR (neoTCR) isolated from circulating T cells of patients. The neoTCRs were isolated using a personalized library of soluble predicted neoantigen–HLA capture reagents. Sixteen patients with different refractory solid cancers received up to three distinct neoTCR transgenic cell products. Each product expressed a patient-specific neoTCR and was administered in a cell-dose-escalation, first-in-human phase I clinical trial (NCT03970382). One patient had grade 1 cytokine release syndrome and one patient had grade 3 encephalitis. All participants had the expected side effects from the lymphodepleting chemotherapy. Five patients had stable disease and the other eleven had disease progression as the best response on the therapy. neoTCR transgenic T cells were detected in tumor biopsy samples after infusion at frequencies higher than the native TCRs before infusion. This study demonstrates the feasibility of isolating and cloning multiple TCRs that recognize mutational neoantigens. Moreover, simultaneous knockout of the endogenous TCR and knock-in of neoTCRs using single-step, non-viral precision genome-editing are achieved. The manufacture of neoTCR engineered T cells at clinical grade, the safety of infusing up to three gene-edited neoTCR T cell products and the ability of the transgenic T cells to traffic to the tumors of patients are also demonstrated

Foy, Susan P., et al. “Non-Viral Precision T Cell Receptor Replacement for Personalized Cell Therapy.” Nature, 2022, https://doi.org/10.1038/s41586-022-05531-1.

If you have any questions about the Ultra-Sensitive Streptavidin Coated Clear 96 Well Microplate or our other offerings, please contact us here.

The Eagle Bioscience’s Pregnenolone ELISA Assay was utilized in a recent publication that explored the effects of pregnenolone on provoked alcohol cravings, anxiety, HPA axis and autonomic arousal in individuals with alcohol use disorder. Check out the full text and abstract below.

Abstract

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The Eagle Bioscience’s Histamine ELISA Assay was highlighted in a recent publication that explored how histamine deficiency deteriorates LPS-induced periodontal diseases in mice. Check out the full text and abstract below.

Abstract

Histamine is a versatile biogenic amine, generated by the unique enzyme histidine decarboxylase (Hdc). Accumulating evidence has proven that histamine plays important roles in numerous biological and pathophysiological processes. However, the role and mechanism of Hdc/Histamine signaling in periodontal diseases remain unclear. In our current study, the concentration of histamine increased in the serum, and Hdc gene expression was upregulated in the gingiva of WT mice with LPS-induced periodontal inflammation. With Hdc–GFP mice, we identified that Hdc/GFP in the periodontium was expressed in CD11b⁺ myeloid cells, rather than in tryptase-positive mast cells. Hdc-expressing CD11b⁺Gr-1⁺ neutrophils significantly increased in the peripheral blood of Hdc–GFP mice one day after LPS injection. Lack of histamine in Hdc^-/- mice not only promoted the activation and infiltration of more CD11b⁺ cells into the peripheral blood but also upregulated mRNA expression levels of IL-1β, IL-6, MCP-1and MMP9 in the gingiva compared to WT mice one day after LPS stimulation. 28 days after LPS treatment, we observed that Hdc^-/- mice exhibited more alveolar bone loss and more osteoclasts than WT mice, which was slightly ameliorated by the administration of exogenous histamine. In vivo and in vitro mechanistic studies revealed that the mRNA expression levels of proinflammatory cytokines and protein levels of NLRP3, Caspase-1, and cleaved-Caspase-1 were upregulated after blocking histamine receptor 1 and 2, especially histamine receptor 1. Taken together, CD11b⁺Gr-1⁺ neutrophils are the predominant Hdc-expressing sites in periodontal inflammation, and deficiency of endogenous histamine in Hdc^-/- mice exacerbates the destruction of the periodontium. Disruption of the histamine/H₁R/H₂R axis aggravates the inflammatory immune response via NLRP3/Casapse-1 pathway.

Song, Fujie., et al. “Histamine Deficiency Deteriorates LPS-Induced Periodontal Diseases in a Murine Model via NLRP3/Caspase-1 Pathway.” International Immunopharmacology, vol. 115, 2023, p. 109630., https://doi.org/10.1016/j.intimp.2022.109630.

If you have any questions about our Histamine ELISA Assay or our other offerings, please contact us here.

The Eagle Bioscience’s 25-OH Vitamin D ELISA Assay Kit has been used in a number of recent publications! These studies range from exploring the effects of vitamin D on cellular responses, molecular immunity, and mycobacterial killing in cattle, to the role of vitamin D, DKK1, hepcidin, and other oxidative stress biomarkers in type 2 diabetes mellitus patients. Check them all out below!

Flores Villalva, Susana. “The Effects of Vitamin D on the Cellular Responses, Molecular Immunity, and Mycobacterial Killing in Cattle.” University College Dublin. School of Agriculture and Food Science, 2022.

Kamel, Amira A., et al. “The Role of Vitamin D, DKK1, Hepcidin and Oxidative Stress Biomarkers in Type 2 Diabetes Mellitus Patients with and without Diabetic Nephropathy.” The Egyptian Journal of Hospital Medicine, vol. 89, no. 2, 2022, pp. 7137–7146.

Blakely, L.P., et al. “Effect of Vitamin D Source and Amount on Vitamin D Status and Response to Endotoxin Challenge.” Journal of Dairy Science, vol. 106, no. 2, 2023, pp. 912–926.

Fernández-Lázaro, Diego, et al. “25-Hydroxyvitamin D Serum Levels Linked to Single Nucleotide Polymorphisms (Snps) (RS2228570, RS2282679, rs10741657) in Skeletal Muscle Aging in Institutionalized Elderly Men Not Supplemented with Vitamin D.” International Journal of Molecular Sciences, vol. 23, no. 19, 2022, p. 11846.

Kumar, Abhai, et al. “Vitamin D and Inflammatory Cytokines Association in Mild Cognitive Impaired Subjects.” Neuroscience Letters, vol. 795, 2023, p. 137044.

Stenhouse, Claire, et al. “Uptake of Phosphate, Calcium, and Vitamin D by the Pregnant Uterus of Sheep in Late Gestation: Regulation by Chorionic Somatomammotropin Hormone.” International Journal of Molecular Sciences, vol. 23, no. 14, 2022, p. 7795.

If you have any questions about our 25-OH Vitamin D ELISA Kit or any of our other offerings, contact us here.

The Eagle Bioscience’s Plasma Metanephrine ELISA Assay was utilized in a recent publication that focused on how social isolation exacerbates acute ozone inhalation induced pulmonary and systemic health outcomes . Check out the full text and abstract below.

Abstract

Psychosocially-stressed individuals might have exacerbated responses to air pollution exposure. Acute ozone exposure activates the neuroendocrine stress response leading to systemic metabolic and lung inflammatory changes. We hypothesized chronic mild stress (CS) and/or social isolation (SI) would cause neuroendocrine, inflammatory, and metabolic phenotypes that would be exacerbated by an acute ozone exposure. Male 5-week-old Wistar-Kyoto rats were randomly assigned into 3 groups: no stress (NS) (pair-housed, regular-handling); SI (single-housed, minimal-handling); CS (single-housed, subjected to mild unpredicted-randomized stressors [restraint-1 h, tilted cage-1 h, shaking-1 h, intermittent noise-6 h, and predator odor-1 h], 1-stressor/day*5-days/week*8-weeks. All animals then 13-week-old were subsequently exposed to filtered-air or ozone (0.8-ppm) for 4 h and immediately necropsied. CS, but not SI animals had increased adrenal weights. However, relative to NS, both CS and SI had lower circulating luteinizing hormone, prolactin, and follicle-stimulating hormone regardless of exposure (SI > CS), and only CS demonstrated lower thyroid-stimulating hormone levels. SI caused more severe systemic inflammation than CS, as evidenced by higher circulating cytokines and cholesterol. Ozone exposure increased urine corticosterone and catecholamine metabolites with no significant stressor effect. Ozone-induced lung injury, and increases in lavage-fluid neutrophils and IL-6, were exacerbated by SI. Ozone severely lowered circulating thyroid-stimulating hormone, prolactin, and luteinizing hormone in all groups and exacerbated systemic inflammation in SI. Ozone-induced increases in serum glucose, leptin, and triglycerides were consistent across stressors; however, increases in cholesterol were exacerbated by SI. Collectively, psychosocial stressors, especially SI, affected the neuroendocrine system and induced adverse metabolic and inflammatory effects that were exacerbated by ozone exposure.

Henriquez, Andres R., et al. “Social Isolation Exacerbates Acute Ozone Inhalation Induced Pulmonary and Systemic Health Outcomes.” Toxicology and Applied Pharmacology, vol. 457, 2022, p. 116295., https://doi.org/10.1016/j.taap.2022.116295.

If you have any questions about the Plasma Metanephrine ELISA Assay or our other offerings, please contact us here.