Author: Eagle Biosciences

The Eagle Bioscience’s ASCA IgG ELISA Kit was highlighted in a recent publication. The aim of this study was to validate an enzyme-linked immunosorbent assay (ELISA) for detection of anti-Saccharomyces cerevisiae antibodies (ASCA) in diabetic patients with foot ulcers, after treatment. Check out the abstract and full article below.

Abstract

This work describes the validation of an enzyme-linked immunosorbent assay (ELISA) for detection of anti-Saccharomyces cerevisiae antibodies (ASCA) in diabetic patients with foot ulcers, after the treatment with Heberprot-P®. Validation followed regulatory guidelines of US FDA and European Medicine Agency. Minimum required dilution of samples and quality controls were defined using pools of sera from diabetic patients and from healthy donors. Parameters such as cut point, specificity, precision, selectivity, robustness and sample stability were analyzed. The repeatability and intermediate precision percent ranged between 7.93-10.61% and 7.93-11.43 %, respectively, indicating low intra- and inter-assay variation. The specificity was proved by background noise suppression, reaching 100% of inhibition as strong criterion for the specificity of the immunoassay. The validated ELISA is a reliable tool for ASCA detection in human serum after the administration of Heberprot-P®, in order to find immunological reactions associated with latent contamination by host cell proteins from Saccharomyces cerevisiae.

Perez-Bernal M., Hernandez C., Delgado M., et al. ELISA validation approach for the detection of anti-saccharomyces cerevisiae antibodies in patients treated with biopharmaceutical heberprot-P. J. Anal. Pharm. Res. (2021) 10(2):50-56.

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The Eagle Bioscience’s Serotonin ELISA Assay Kit was highlighted in a recent publication on the selective, ultra-sensitive, and rapid detection of serotonin by an optimized ZnO Nanorod FET biosensor. Check out the abstract and full article below.

Abstract

Background: Fluctuation in serotonin (5-HT) level is an essential manifestation of several neurological disorders. In view of such importance, it is necessary to monitor the levels of 5-HT with good sensitivity, selectivity, affordability and low response time. Zinc oxide (ZnO) based field effect transistors (FET) with attributes like minimized noise levels and large on-off ratio are regarded as emerging high performance biosensor platforms. However, their response is significantly non-linear and there has been no appreciable endeavor for improving the non-linearity. Method: In this paper, we have introduced embedded gate electrode encompassing the channel of the FET which improves the uniformity in electric field line distribution through the electrolyte and proportionately enhances the capture of target biomolecule at ultra-low concentrations, thereby increasing the linearity. Further, we have incorporated the optimized parameters of ZnO nanorods reported previously, for rapid and selective detection of 5-HT. Results: It has been observed that the fabricated ZnO FET biosensor lowers the detection limit down to 0.1fM which is at least one order of magnitude lower than the existing reports. The sensor also has wide linear range from 0.1fM to 1nM with a detection time of about 20 minutes. Conclusion: The proposed zinc oxide nanorod-based sensor can be used as an excellent tool for future diagnosis of neurological disorders.

Sinha K., Chakraborty B., Chaudhury S.S., et al. Selective, Ultra-sensitive and Rapid Detection of Serotonin by Optimized ZnO Nanorod FET Biosensor. IEEE Transactions on NanoBioscience.

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The Eagle Bioscience’s Noradrenaline Sensitive ELISA was highlighted in a recent publication on how brain injuries instruct bone marrow cellular lineage destination to reduce neuroinflammation. Check out the abstract and full article below.

Abstract

Acute brain injury mobilizes circulating leukocytes to transmigrate into the perivascular space and brain parenchyma. This process amplifies neural injury. Bone marrow hematopoiesis replenishes the exhausted peripheral leukocyte pools. However, it is not known whether brain injury influences the development of bone marrow lineages and how altered hematopoietic cell lineages affect neurological outcome. Here, we showed that bone marrow hematopoietic stem cells (HSCs) can be swiftly skewed toward the myeloid lineage in patients with intracerebral hemorrhage (ICH) and experimental ICH models. Lineage tracing revealed a predominantly augmented hematopoiesis of Ly6Clow monocytes infiltrating the ICH brain, where they generated alternatively activated macrophages and suppressed neuroinflammation and brain injury. The ICH brain uses β3-adrenergic innervation that involves cell division cycle 42 to promote bone marrow hematopoiesis of Ly6Clow monocytes, which could be further potentiated by the U.S. Food and Drug Administration-approved β3-adrenergic agonist mirabegron. Our results suggest that brain injury modulates HSC lineage development to curb distal brain inflammation, implicating the bone marrow as a unique niche for self-protective neuroimmune interaction that might be exploited to obtain therapeutic effects.

Shi, SX., Shi, K., Liu, Q. Brain injury instructs bone marrow cellular lineage destination to reduce neuroinflammation. Science Translational Medicine. (2021)13:589.

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The Eagle Bioscience’s Glutathione Total Assay Kit was highlighted in a recent publication on the protective effects of perindopril against indomethacin-induced gastric mucosal damage through modulation of DDAH-1/ADMA and ACE-2/ANG-(1-7) signaling pathways. Check out the abstract and full article below.

Abstract

Indomethacin is a widely used nonsteroidal anti-inflammatory drug; however, its clinical utility is accompanied by serious adverse reactions including peptic ulcers. The current study aims to investigate the protective potential of perindopril against indomethacin-induced gastric injury in rats. Perindopril (4 mg/kg) was administered orally for 7 days and indomethacin (60 mg/kg, single oral dose) was administered on the 7th day, 1 h after perindopril administration. Pantoprazole was used as a standard agent. Ulcer index (UI), preventive index ratio (PI), histopathological examination, oxidative stress, and inflammatory biomarkers were investigated. Perindopril significantly decreased UI while increased PI and counteracted histopathological aberrations induced by indomethacin. It alleviated indomethacin-induced oxidative stress by lowering NO while increasing GSH content and superoxide dismutase activity. Perindopril significantly downregulated TNF-α and asymmetric dimethylarginine (ADMA), while significantly upregulated COX-2, PGE-2, dimethylarginine dimethylaminohydrolase-1 (DDAH-1), ANG-(1-7), and ACE-2 expression. Together, these findings suggest the gastroprotective effects of perindopril through modulation of DDAH-1/ADMA and ACE-2/ANG-(1-7) signaling.

HIGHLIGHTS

• Perindopril attenuated gastric histopathological damage.
• It increased GSH content and SOD activity while decreased NO content.
• It modulated gastric ADMA and DDAH-1 activity.
• It reduced TNF-α, while increased COX-2 and PGE-2 expression.
• It upregulated ACE-2 activity and ANG-(1-7) protein expression.

Mohamed Y.T., Naguib I.A., Abo-Saif A.A., Mohamed W.R. Protective effects of perindopril against indomethacin-induced gastric mucosal damage through modulation of DDAH-1/ADMA and ACE-2/ANG-(1-7) signaling pathways. Drug Chem. Toxicol. (2021).

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The Eagle Bioscience’s Coronavirus COVID-19 IgG ELISA Assay was recently highlighted in a publication on the decline of Sars-CoV-2 antibodies over 6-month follow-up in obstetrical healthcare workers (HCW). Researchers investigated the longitudinal presence of serum Sars-CoV-2 specific antibodies for both Immunoglobulin G (IgG) and Immunoglobulin M (IgM) in obstetrical HCWs at a tertiary hospital. Check out the full article below.

Abstract

The Problem

Limited data exists on the temporal trend of the Sars-CoV-2 immunologic response and duration of protection following natural infection. We sought to investigate the presence and duration of Sars-CoV-2 serum antibodies in obstetrical healthcare workers (HCW) on serial assessments over a 6-month period, and to assess rates of vaccine acceptance and reported vaccine side effects among this cohort.

Method of Study

A prospective cohort study of a convenience sample of obstetrical HCWs at a tertiary hospital. Serum Sars-CoV-2 antibodies for Immunoglobulin G (IgG) and Immunoglobulin M (IgM) were measured longitudinally at four intervals: baseline, 4 weeks, 12 weeks, and 6 months. Participants completed voluntary surveys on COVID19 testing, high-risk exposures, vaccine acceptance, and vaccine side effects.

Results

One hundred twenty-six of 150 (84%) HCWs who volunteered for participation completed all four blood draws. Prevalence of seropositive HCWs based on positive Sars-CoV-2 IgG antibodies increased from 2% at baseline to 31% at 12 weeks but declined to 21% by 6 months. Forty-two percent (19/43) of the participants considered seropositive for Sars-CoV-2 IgG antibodies at any of the initial three blood draws converted to seronegative status at the 6-month follow-up. Eighty-seven percent (72/83) of participants who responded to a follow-up survey were willing to accept the COVID19 vaccine. Rates of acceptance did not differ by participant antibody status. Those that experienced symptoms with the first injection were more likely to have positive Sars-CoV-2 IgG antibodies (36.8% vs. 9.6%, p = .01).

Conclusion

Sars-CoV-2 IgG antibodies wane over time and may not provide prolonged and robust immune protection. This underscores the importance of vaccination and continued research in this area while the COVID19 pandemic continues.

Kiefer M.K., Allen K.D., Russo J.R., et al. Decline in Sars-CoV-2 antibodies over 6-month follow-up in obstetrical healthcare workers. Am. J. Reprod. Immunol. (2021)

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The Eagle Bioscience’s MedFrontier Intact FGF23 Assay was utilized in a recent publication to determine intestinal phosphorus absorption in patients with moderate chronic kidney disease (CDK) and healthy adults. Find the abstract and full text below.

Abstract

Reducing intestinal phosphorus absorption is a cornerstone in CKD-MBD management. Yet, knowledge gaps include how CKD pathophysiology affects intestinal phosphorus absorption. In vivo rodent studies suggest that intestinal phosphorus absorption remains inappropriately normal in early-moderate CKD, despite declining 1,25-dihydroxyvitamin D (1,25D). We measured intestinal phosphorus absorption in patients with moderate CKD versus healthy adults using a direct radiotracer method. Patients with CKD and healthy adults matched for age, sex, and race were enrolled in this 8-day controlled diet study: the first 6 days outpatient and the final 2 days inpatient. Oral and intravenous doses of 33P and serial blood and urine sampling determined intestinal phosphorus absorption during the final 2 days. Secondary outcomes included fasting biochemistries and 24-hour urine phosphorus (uP). In total, n=8 patients with CKD (eGFR=29-55 ml/min per 1.73 m2) and n=8 matched healthy controls completed the study. On a controlled diet, no difference in fractional intestinal phosphorus absorption was detected between patients with CKD and healthy adults (0.69 versus 0.62, respectively; P=0.52), and this was similar for 24-hour uP (884 versus 935 mg/d, respectively; P=0.70). Fractional intestinal phosphorus absorption was not significantly related to 24-hour uP. Patients with CKD had higher serum intact PTH and intact FGF23 and lower 1,25D. The relationship between 1,25D and fractional intestinal phosphorus absorption was not statistically significant. Intestinal phosphorus absorption with typical dietary intake did not differ in patients with moderate CKD compared with controls, despite lower serum 1,25D levels. In this setting, a relationship between 24-hour uP and fractional or absolute intestinal absorption was not evident. Further investigation is needed to determine what factors influence intestinal phosphorus absorption in CKD and the apparent lack of compensation by the intestine to limit phosphorus absorption in the face of declining kidney function and reduced 1,25D. Whether this is evident across a range of dietary phosphorus intakes, as well as CKD severity, also needs to be determined.

Stremke E.R, Wises G.N., Moe S.M., et al. Intestinal Phosphorus Absorbtion in Moderate CKD and Healthy Adults Determined Using a Radioisotopic Tracer. J. Am. Soc. Nephrol. (2021) 32(8):2057-2069

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The Eagle Bioscience’s Dopamine Sensitive ELISA Assay was utilized in a recent publication on how Hexachloronaphthalene (HxCN) impairs the dopamine pathway in an in vitro model of PC12 cells. This is the first study to demonstrate that HxCN administered in vitro inhibits dopamine biosynthesis and secretion through the regulation of tyrosine hydroxylase and VMAT1 expression. Check out the abstract and full article below.

Abstract

Among polychlorinated naphthalenes (PCNs), listed by the Stockholm convention as Persistent Organic Pollutants (POPs), hexachloronaphthalenes are considered the most toxic and raise the highest concern. Of these, 1,2,3,5,6,7-hexachloronaphthalanene (PCN67) is considered the main congener affecting human health due to its hepatotoxicity and its ability to disturb the reproductive, endocrine, and hematological systems. It is also prevalent in human serum/plasma, milk, and adipose tissue. However, little is known about its neurotoxicity, despite the fact that anorectic effects have been observed in workers occupationally exposed to PCNs and in animal research on PCN67. Since dopamine is involved in many aspects of food intake, the aim of this study was to confirm whether PCN67 affects dopamine synthesis in differentiated PC12 cells, a widely used model of neurosecretion. Our results show that exposure to PCN67 resulted in diminished dopamine content and release. Moreover, PCN67 also affected the expression of tyrosine hydroxylase and lowered the expression of vesicular monoamine transporter 1 (VMAT1). In addition, significantly lower expression of antioxidant enzymes, including catalase, glutathione peroxidase and copper/zinc superoxide dismutase, was observed in comparison to the vehicle. In conclusion, PCN67 appears to disturb dopaminergic transmission by altering tyrosine hydroxylation, reducing VMAT1 expression and impairing antioxidant protection. Our study provides a potential mechanism for how PCN67 may cause dopamine deficiency and contribute to neuronal death by affecting cellular antioxidant potency; however, this conclusion requires further research.

Lisek M., Boczek T., Stragierowicz J., et al. Hexachloronaphthalene (HxCN) impairs the dopamine pathway in an in vitro model of PC12 cells. Chemosphere. (2021) 287:3.

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The Eagle Bioscience’s Anti-Beta2 Glycoprotein 1 IgG ELISA was recently highlighted in a publication on the effects of hydroxychloroquine onantiphospholipid antibodies-inhibited endometrial angiogenesis. Check out the abstract and full-text article below.

Abstract

Antiphospholipid syndrome (APS) is an autoimmune disease characterized by thrombotic events and/or pregnancy morbidity (≥3 recurrent early miscarriage or fetal death or a prematurity <34 weeks of gestation) with persistently positive antiphospholipid antibodies (aPLs). It is reported that aPLs damage the placental tissue by binding to β2-glycoprotein I (β2GPI) on the surface of trophoblast and endothelial cells. Hydroxychloroquine (HCQ) is considered to be beneficial in the treatment of obstetrical APS and shown to restore the aPL-inhibited invasion and differentiation of trophoblast. However, not enough evidence exists regarding the effect of HCQ on endometrial angiogenesis. The aim of our study was to assess whether HCQ has an effect on aPL-inhibited endothelial angiogenesis. In this research, to explore the effect of HCQ for angiogenesis, we investigated: Human umbilical vein endothelial cells (HUVECs) viability by CCK-8; HUVECs migration by wound healing; HUVEC angiogenesis by Matrigel assay in vitro; mRNA expression of MMP-2 and VEGF by real-time quantitative Polymerase Chain Reaction (RT-PCR); protein expression of VEGF, MMP-2 by western blot. We found that HCQ treatment significantly restored the expression of aPL-inhibited VEGF and MMP-2. HCQ restored aPL-inhibited HUVEC proliferation, migration, and angiogenesis in vitro. In conclusion, aPLs inhibit HUVECs angiogenesis, however, HCQ can restore the effect of aPL-inhibited HUVECs migration and angiogenesis in vitro, demonstrating its beneficial therapeutic role in obstetrical APS.

Dong Y., Lu Y., Xia Y., Wang X. Effect of hydroxychloroquine on antiphospholipid antibodies inhibited endometrial angiogenesis. J. Matern. -Fetal Neonatal Med. (2021).

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The Eagle Bioscience’s 25-OH Vitamin D ELISA Assay was highlighted in a recent publication by the Department of Oncology at Gerogetown Univeristy in DC. Researchers determined how the vitamin D receptor is linked to disease outcome in estrogen receptor-positive (ER+) breast cancer patients treated with tamoxifen (TAM). Check out the abstract and full text article below.

Abstract

We determined how vitamin D receptor (VDR) is linked to disease outcome in estrogen receptor-positive (ER+) breast cancer patients treated with tamoxifen (TAM). Breast cancer patients (n = 581) in four different datasets were divided into those expressing higher (above median) and lower levels of VDR in pretreatment ER+ tumors. Across all datasets, TAM-treated patients with higher pretreatment tumor VDR expression exhibited significantly longer recurrence-free survival. Ingenuity pathway analysis identified autophagy and unfolded protein response (UPR) as top differentially expressed pathways between high and low VDR-expressing ER+ cancers. Activation of VDR with vitamin D (VitD), either calcitriol or its synthetic analog EB1089, sensitized MCF-7-derived, antiestrogen-resistant LCC9 human breast cancer cells to TAM, and attenuated increased UPR and pro-survival autophagy. Silencing of VDR blocked these effects through the IRE1α-JNK pathway. Further, silencing of VDR impaired sensitivity to TAM in antiestrogen-responsive LCC1 cells, and prevented the effects of calcitriol and EB1089 on UPR and autophagy. In a preclinical mouse model, dietary VitD supplementation induced VDR activation and reduced carcinogen-induced ER+ mammary tumor incidence. In addition, IRE1α-JNK signaling was downregulated and survival autophagy was inhibited in mammary tumors of VitD-supplemented mice. Thus, activation of VDR is predictive of reduced risk of breast cancer recurrence in ER+ patients, possibly by inhibiting antiestrogen-promoted pro-survival autophagy.

Li, Y., Cook, KL., Yu, W., et al. Inhibition of Antiestrogen-Promoted Pro-Survival Autophagy and Tamoxifen Resistance in Breast Cancer through Vitamin D Receptor. Nutrients. (2021) 13(5):1715

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