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The Eagle Bioscience’s Folic Acid ELISA Assay Kit Highlighted in a recent study! The study was conducted to assess the disruption in nutritional attributes of Capsicum annuum by five different microplastic (MP) types. Check out the abstract and full text below.


Abstract

This study was executed to assess the disruption in nutritional attributes of Capsicum annuum by five different microplastic (MP) types i.e. polyvinylchloride (PVC), polystyrene (PS), high-density polyethylene (HDPE), low-density polyethylene (LDPE), and polyethylene terephthalate (PET). Significant (P ≤ 0.05) differences were recorded among traits of C. annuum fruit concerning the type of MPs. PVC was more hazardous than the other MPs and depleted maximum protein content. Likewise, HDPE exerted a 51.62% reduction in carbohydrates compared to the control. Vitamin-A (-32.09%) and vitamin-B6 (-37%) were severely influenced by the PVC in C. annuum fruit. HDPE and PVC both declined the oleic acid contents. PVC caused a 47.6% reduction in linoleic acid while the least damaging was the PET (4.71%) for this attribute. Palmitic acid and stearic acid were however more negatively affected by the HDPE. The PVC and HDPE severely degraded the total flavonoid contents and phenolics. Macro and micronutrients of C. annuum fruit were also negatively influenced by all the five MPs used in this study. HDPE, PVC and LDPE, respectively caused significant repression in Ca, K, Mg and Zn (P ≤ 0.05). Overall, HDPE and PVC caused significant damage and decreased nutritional contents. This is, so far, the premier study unraveling the changes in the nutrition of C. annuum fruits due to MPs. Accordingly, further research is highly recommended on the changes caused by MPs in the nutritional value of food crops.

Khadiga Alharbi, Muhammad Aqeel, Noreen Khalid, Atia Nazir, Muhammad Kashif Irshad, Fahad Mohammed Alzuaibr, Haifa AbdulAziz Sakit AlHaithloul, Noreen Akhter, Omar Mahmoud Al-Zoubi, Muhammad Qasim, Khalid M.Al Syaad, Manal Abdullah AlShaqhaa, Ali Noman. Microplastics in soil differentially interfere with nutritional aspects of chilli peppers. South African Journal of Botany. Volume 160. 2023. Pages 402-413. ISSN 0254-6299, https://doi.org/10.1016/j.sajb.2023.07.027.


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What is it SpheroTribe intended for?

SpheroTribe provides a simple toolkit to generate consistent and robust 3D cell structures. Simply dilute the SpheroTribe solution into your culture medium of choice, watch your cells turn into uniformly sized 3D spheroids and collect them for your downstream assays.

Once diluted in your culture medium of choice, our concentrated polymer-based solution increases the medium viscosity favoring cell-cell contacts. SpheroTribe offers a simple method to generate homogeneous 3D cell structures with increased control over their size and shape, which can be easily handled and washed for downstream experiments.

SpheroTribe is particularly useful to boost aggregation when working with challenging cells, minimize variability between samples and improve the consistency of your migration/invasion assays, immunostaining, drug screening or in vivo implantation experiments.

SpheroTribe improves cardiac organoid formation by boosting hiPSC aggregation

U-87 glioblastoma cells were seeded at 1,000 cells per well in round-bottom wells molded in agarose using a Stampwell U-shape (Idylle) in full culture medium without (right) or added with SpheroTribe (left). After 3 days, pictures were taken and number of cell aggregates per well were quantified from 64 independent wells and associated standard deviation (SD) values were calculated.


Kit Description:

You can purchase just the methylcellulose solution or choose among 2 different size kits:

25mL kit contents:

  • 25mL of 5X methylcellulose solution
  • 10x U-bottom 96-well plates
  • 2x racks of 96 pipette tips (200µL) with a large opening
2.5mL kit contents:

    • 2.5mL of 5X methylcellulose solution
    • 1x U-bottom 96-well plate
    • 20 pipette tips (200µL) with a large opening

Applications

SpheroTribe has been successfully used for spheroid/organoid formation with the following cell types:
Patient-derived stem-like glioblastoma cells (GB P3 and BL13), human glioblastoma cell lines (U87 & T98G), HeLa, human vaginal mucosal melanoma (HMV-II), human primary colorectal cancer cells, human breast cancer cells (MDA-MB 231), human induced pluripotent stem cells, monkey kidney fibroblast-like cell line (COS-7), primary neurons from rat embryos (E18) & murine melanoma cells (B16F10).

Experimental assays:
Once spheroids have grown to your desired size, you can use them for any kind of assay according to your regular workflow. The SpheroTribe solution can be readily washed off, leaving a spheroid available for other tests at any stage of your protocol.

Immune infiltration of B16F10 spheroids after immune checkpoint blockade

A. 10,000 B16F10 cells were grown for 6 days as spheroids using SpheroTribe. B. 100,000 PBMC from murine spleen were activated with IL-15 (40 ng/mL) [1], incubated with anti-PD1 (10 µg/mL) for 1h and added on B16F10 spheroids for 3 days.
Graph shows flow cytometry quantification of differential lymphocyte infiltration after spheroid dissociation according to treatment. N=4. Mann-Whitney U Test, p-value<0.05. [1] https://doi.org/10.3389/fonc.2022.898732.


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The Eagle Bioscience’s Dopamine ELISA Assay Kit was utilized in a recent publications! The aim of the study was to investigate the potential of iron oxide nanoparticles produced using ascorbic acid (AA-IONPs) against Parkinson’s Disease. Check out the abstract and full text below!


Abstract

One of the most prevalent neurological movement diseases affecting the geriatric population globally is Parkinson’s disease (PD). Recent studies have highlighted the potency of biomolecules in the generation of nanomaterials and also over their impact on neuroprotection. The objective of this research was to investigate the potential of iron oxide nanoparticles produced using ascorbic acid (AA-IONPs) against PD. Numerous analytical methods including UV–Vis analysis, Fourier-Transform Infrared Spectroscopy (FTIR), dynamic light scattering (DLS), and electron microscopy (SEM, TEM), were used to analyze the produced AA-IONPs. Nitric oxide, prostaglandin E2, and inflammatory cytokines analyses such as IL-6 and IL-1 were employed to assess the neuroprotective effect of synthesized AA-IONPs on inflammatory agent lipopolysaccharides driven murine microglial BV2 cells. And also Parkinson-induced C57BL/6 mice were given the nanoparticle treatment to confirm the in vivo effects of the produced nanoparticles. Our characterization findings had demonstrated that AA-IONPs have a significant role in acting as an ideal nano drug and may have the ability to reduce inflammation in in vitro murine microglial BV2. The outcomes of in vivo tests conclusively show that AA-IONPs had reduced neuroinflammation and enhanced motor coordination in Parkinson’s disease-induced rats.

Li, L., Luo, P., Wu, S. et al. Deciphering the neuroprotective effect of ascorbic acid mediated synthesis of iron oxide nanoparticles against Parkinson’s disease: an in vitro and in vivo approach. Macromol. Res. 31, 949–960 (2023). https://doi.org/10.1007/s13233-023-00186-x


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The Biomedica Human IL-6 High Sensitive ELISA Assay Kit was highlighted in a recent publication that explored factors associated with incident vertebral fractures in glucocorticoid-treated Duchenne muscular dystrophy. Check out the abstract and full text below!


Abstract

Purpose: Prevention of fractures is an unmet need in glucocorticoid (GC)-treated Duchenne muscular dystrophy. This study explored factors associated with incident vertebral fractures (VFs) to inform future fracture prevention efforts.

Methods: VFs were evaluated prospectively at study baseline and 12 months on lateral spine radiographs in participants aged 4 to 25 years with Duchenne muscular dystrophy. Clinical factors were analyzed for their association with the change in Spinal Deformity Index (sum of the Genant-defined VF grades from T4 to L4) between baseline and 12 months.

Results: Thirty-eight males were evaluated (mean ± SD age at baseline 11.0 ± 3.6 years; mean ± SD GC duration at baseline 4.1 ± 3.1 years; 74% ambulatory). Nine of 38 participants (24%) had 17 incident VFs, of which 3/17 VFs (18%) were moderate/severe. Participants with 12-month incident VF had lower mean ± SD baseline lumbar spine areal bone mineral density Z-scores (-2.9 ± 1.0 vs -1.9 ± 1.1; P = .049) and lower total body less head areal bone mineral density Z-scores (-3.1 ± 1.2 vs -1.6 ± 1.7; P = .036). Multivariable linear regression showed that at least 1 VF at baseline (P < .001), a higher number of antecedent non-VF (P < .001), and greater bone age delay at baseline (P = .027) were significant predictors of an increase in the Spinal Deformity Index from baseline to 12 months.

Conclusion: The observation that ≥ 1 prevalent VF and/or non-VF were the strongest predictors of incident VFs at 12 months supports the need for prevention of first fractures in this high-risk setting. Bone age delay, a marker of GC exposure, may assist in the prioritization of patients in efforts to prevent first fractures.

Keywords: Duchenne muscular dystrophy; bone fragility; glucocorticoids; incident fractures; osteoporosis; vertebral fractures.

Risk Factors Associated with Incident Vertebral Fractures in Steroid-treated Males with Duchenne Muscular Dystrophy . Phung K, McAdam L, Ma J, McMillan HJ, Jackowski S, Scharke M, Matzinger MA, Shenouda N, Koujok K, Jaremko JL, Wilson N, Walker S, Hartigan C, Khan N, Page M, Robinson ME, Saleh DS, Smit K, Rauch F, Siminoski K, Ward LMJ Clin Endocrinol Metab. 2023 Aug 23:dgad435. doi: 10.1210/clinem/dgad435. Epub ahead of print.PMID: 37610420


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Dopamine Sensitive ELISA Assay Utilized in Recent Publication

The Eagle Bioscience’s Noradrenaline ( Norepinephrine) High Sensitive ELISA was utilized in a recent publication that focused on how β1-adrenergic receptor links sympathetic nerves to T cell exhaustion. Check out the full text and abstract below!


Abstract

CD8+ T cells are essential components of the immune response against viral infections and tumors, and are capable of eliminating infected and cancerous cells. However, when the antigen cannot be cleared, T cells enter a state known as exhaustion1. Although it is clear that chronic antigen contributes to CD8+ T cell exhaustion, less is known about how stress responses in tissues regulate T cell function. Here we show a new link between the stress-associated catecholamines and the progression of T cell exhaustion through the β1-adrenergic receptor ADRB1. We identify that exhausted CD8+ T cells increase ADRB1 expression and that exposure of ADRB1+ T cells to catecholamines suppresses their cytokine production and proliferation. Exhausted CD8+ T cells cluster around sympathetic nerves in an ADRB1-dependent manner. Ablation of β1-adrenergic signaling limits the progression of T cells towards the exhausted state in chronic infection and improves effector functions when combined with immune checkpoint blockade (ICB) in melanoma. In a pancreatic cancer model resistant to ICB, β-blockers and ICB synergize to boost CD8+ T cell responses and induce the development of tissue-resident memory-like T cells. Malignant disease is associated with increased catecholamine levels in patients2,3, and our results establish a connection between the sympathetic stress response, tissue innervation and T cell exhaustion. Here, we uncover a new mechanism by which blocking β-adrenergic signaling in CD8+ T cells rejuvenates anti-tumor functions.

Globig, AM., Zhao, S., Roginsky, J. et al. The β1-adrenergic receptor links sympathetic nerves to T cell exhaustion. Nature 622, 383-392 (2023).


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

Dopamine Sensitive ELISA Assay Utilized in Recent Publication

The Eagle Bioscience’s easYmer H2-Db MHC Tetramer was highlighted in a recent publication that focused on killing tumor-associated bacteria with a liposomal antibiotic and how it generates neoantigens that induce anti-tumor immune responses. Check out the full text and abstract below!


Abstract

Increasing evidence implicates the tumor microbiota as a factor that can influence cancer progression. In patients with colorectal cancer (CRC), we found that pre-resection antibiotics targeting anaerobic bacteria substantially improved disease-free survival by 25.5%. For mouse studies, we designed an antibiotic silver-tinidazole complex encapsulated in liposomes (LipoAgTNZ) to eliminate tumor-associated bacteria in the primary tumor and liver metastases without causing gut microbiome dysbiosis. Mouse CRC models colonized by tumor-promoting bacteria (Fusobacterium nucleatum spp.) or probiotics (Escherichia coliNissle spp.) responded to LipoAgTNZ therapy, which enabled more than 70% long-term survival in two F. nucleatum-infected CRC models. The antibiotic treatment generated microbial neoantigens that elicited anti-tumor CD8+ T cells. Heterologous and homologous bacterial epitopes contributed to the immunogenicity, priming T cells to recognize both infected and uninfected tumors. Our strategy targets tumor-associated bacteria to elicit anti-tumoral immunity, paving the way for microbiome–immunotherapy interventions.

Wang, M., Rousseau, B., Qiu, K. et al. Killing tumor-associated bacteria with a liposomal antibiotic generates neoantigens that induce anti-tumor immune responses. Nat Biotechnol (2023).


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Dopamine Sensitive ELISA Assay Utilized in Recent Publication
The Eagle Bioscience’s Calprotectin ELISA Assay Kit was utilized in a recent publication that explored intestinal microbiome and metabolome signatures in patients with chronic granulomatous disease. Check out the full text and abstract below!


Background

Chronic granulomatous disease (CGD) is caused by defects in any 1 of the 6 subunits forming the nicotinamide adenine dinucleotide phosphate oxidase complex 2 (NOX2), leading to severely reduced or absent phagocyte-derived reactive oxygen species production. Almost 50% of patients with CGD have inflammatory bowel disease (CGD-IBD). While conventional IBD therapies can treat CGD-IBD, their benefits must be weighed against the risk of infection. Understanding the impact of NOX2 defects on the intestinal microbiota may lead to the identification of novel CGD-IBD treatments.

Objective

We sought to identify microbiome and metabolome signatures that can distinguish individuals with CGD and CGD-IBD.

Methods

We conducted a cross-sectional observational study of 79 patients with CGD, 8 pathogenic variant carriers, and 19 healthy controls followed at the National Institutes of Health Clinical Center. We profiled the intestinal microbiome (amplicon sequencing) and stool metabolome, and validated our findings in a second cohort of 36 patients with CGD recruited through the Primary Immune Deficiency Treatment Consortium.

Results

We identified distinct intestinal microbiome and metabolome profiles in patients with CGD compared to healthy individuals. We observed enrichment for Erysipelatoclostridium spp, Sellimonas spp, and Lachnoclostridium spp in CGD stool samples. Despite differences in bacterial alpha and beta diversity between the 2 cohorts, several taxa correlated significantly between both cohorts. We further demonstrated that patients with CGD-IBD have a distinct microbiome and metabolome profile compared to patients without CGD-IBD.

Conclusion

Intestinal microbiome and metabolome signatures distinguished patients with CGD and CGD-IBD, and identified potential biomarkers and therapeutic targets.

Chandrasekaran, Prabha, et al. “Intestinal microbiome and metabolome signatures in patients with chronic granulomatous disease.” Journal of Allergy and Clinical Immunology, 2023.


If you have any questions about the Calprotectin ELISA or our other offerings, contact us here.

Dopamine Sensitive ELISA Assay Utilized in Recent Publication

The Eagle Bioscience’s Serotonin ELISA Assay was utilized in a recent publication that explored the modification of the serotonergic systems and phenotypes by gestational micronutrients. Check out the full text and abstract below.


Abstract

Micronutrients consumed in excess or imbalanced amounts during pregnancy may increase the risk of metabolic diseases in offspring, but the mechanisms underlying these effects are unknown. Serotonin (5-hydroxytryptamine, 5-HT), a multifunctional indoleamine in the brain and the gut, may have key roles in regulating metabolism. We investigated the effects of gestational micronutrient intakes on the central and peripheral serotonergic systems as modulators of the offspring’s metabolic phenotypes. Pregnant Wistar rats were fed an AIN-93G diet with 1-fold recommended vitamins (RV), high 10-fold multivitamins (HV), high 10-fold folic acid with recommended choline (HFolRC), or high 10-fold folic acid with no choline (HFolNC). Male and female offspring were weaned to a high-fat RV diet for 12 weeks. We assessed the central function using the 5-HT2C receptor agonist, 1-(3-chlorophenyl)piperazine (mCPP), and found that male offspring from the HV- or HFolRC-fed dams were less responsive (P < 0.05) whereas female HFolRC offspring were more responsive to mCPP (P < 0.01) at 6 weeks post-weaning. Male and female offspring from the HV and HFolNC groups, and male HFolRC offspring had greater food intake (males P < 0.001; females P < 0.001) and weight gain (males P < 0.0001; females P < 0.0001), elevated colon 5-HT (males P< 0.01; females P < 0.001) and fasting glucose concentrations (males P < 0.01; females P < 0.01), as well as body composition toward obesity (males P < 0.01; females P < 0.01) at 12 weeks post-weaning. Colon 5-HT was correlated with fasting glucose concentrations (males R2=0.78, P < 0.0001; females R2=0.71, P < 0.0001). Overall, the serotonergic systems are sensitive to the composition of gestational micronutrients, with alterations consistent with metabolic disturbances in offspring.

Chen, Vicki, et al. “Modification of the Serotonergic Systems and Phenotypes by Gestational Micronutrients.” Journal of Endocrinology, vol. 257, no. 2, 2023, https://doi.org/10.1530/joe-22-0305.


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

FluoBolt- KLOTHO Fluorescence Immunoassay

High Sensitivity; Single Step Immunoassay for α-KLOTHO in Human Serum and Plasma

Eagle Biosciences is excited to highlight the FloBolt- KLOTHO Fluorescence Immunoassay from Fianostics! This assay detects only α-KLOTHO and does not cross-react with β-KLOTHO. No interference of recombinant FGF-23 with the assay’s signal up to a 100-fold molar excess was monitored. Check out the assay background and highlights below!


Background

α-KLOTHO is expressed in kidney, small intestine, placenta and prostate. The soluble peptide can be found in serum and cerebrospinal fluid. It may play a role in the calcium/ phosphorus homeostasis regulation by e.g. inhibiting active vitamin D synthesis. Further, it is also known as an antiaging-hormone by extending life span by inhibiting insulin/ IGF1 signaling pathway, as experiments in mice showed. KLOTHO is a co-receptor of fibroblast growth factor 23 (FGF-23). Research has investigated association of altered serum KLOTHO levels with chronic kidney disease and failure, renal and hepatocellular carcinomas, osteoporosis or cardiovascular diseases.

α-KLOTHO can be found either as a membrane bound or a secreted form. The membrane bound form consists of 1012 amino acids (aa), starting with a 56 aa long signaling sequence and followed by two glycosyl hydrolase 1 regions (position 57-506 and 515-953). Both glycosyl hydrolase 1 regions lack one essential Glu active site residue. Thus, it is inactive in vivo as a glycosidase although it belongs to the glycosyl hydrolase 1 family. KLOTHO’s secreted isoform, which predominates over the membrane bound form, consists of 549 amino acids (aa). It is produced by alternative splicing and differs from the membrane bound form by aa 550 to 1012 missing.


FluoBolt- KLOTHO Fluorescence Immunoassay Highlights

  • High Sensitivity
  • Single Step Procedure
  • No Washing Steps
  • No Enzyme Substrate required
  • Long Term Stable Signal

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Dopamine Sensitive ELISA Assay Utilized in Recent Publication

The Eagle Bioscience’s Cortisol Saliva ELISA was highlighted in a recent publication that focused on peripheral brain-derived neurotrophic factor (BDNF) and salivary cortisol levels in college students with different levels of academic stress. Check out the full text and abstract below!


Abstract

Introduction

Brain-derived neurotrophic factor (BDNF) is essential for brain physiological processes influencing memory and learning. BDNF levels can be affected by many factors, including stress. Stress increase serum and salivary cortisol levels. Academic stress is of the chronic type. BDNF levels can be measure from serum, plasma or platelets, and there is still no standard methodology, which is relevant to ensure reproducibility and comparability between studies.

Hypothesis

(i) BDNF concentrations in serum show greater variability than in plasma. (ii) In college students with academic stress, peripheral BDNF decreases and salivary cortisol increases.

General Objective

To standardize plasma and serum collection for BDNF levels and to determine whether academic stress affects peripheral BDNF and salivary cortisol levels.

Castillo-Navarrete, Juan-Luis, et al. “Peripheral Brain-Derived Neurotrophic Factor (BDNF) and Salivary Cortisol Levels in College Students with Different Levels of Academic Stress. Study Protocol.” PLOS ONE, vol. 18, no. 2, 2023.


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