Category: Publication Spotlight

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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The Eagle Bioscience’s Mouse Rat 25-OH Vitamin D ELISA was recently highlighted about how the MicroRNA-122 contributes to lipopolysaccharide-induced acute kidney injury via down-regulating the vitamin D receptor in the kidney.

Abstract

Background
Our previous studies showed that vitamin D receptor (VDR) depletion promotes lipopolysaccharide (LPS)-induced acute kidney injury (AKI) in mice, and renal VDR is down-regulated in AKI, but the mechanism of VDR down-regulation is unclear.

Methods
Nutritional vitamin D deficiency was induced by feeding mice a vitamin D-deficient (VD-D) diet. Mice were injected intraperitoneally with LPS (20 mg/kg) to establish LPS-induced AKI. Levels of VDR and miR-122 were measured both in vivo and in vitro. The associations between VDR and miR-122 were analysed by dual-luciferase reporter assays.

Results
Compared with vitamin D-sufficient (VD-S) mice, VD-D mice developed more severe renal injury following LPS challenge. LPS induced a dramatic decrease in VDR expression and marked induction of miR-122 both in vivo and in vitro. Furthermore, miR-122 hairpin inhibitor alleviated LPS-induced VDR down-regulation whereas miR-122 mimic directly suppressed VDR expression in HK-2 cells. In luciferase reporter assays, miR-122 mimic was able to suppress luciferase activity in 293T cells co-transfected with a luciferase reporter that contains a putative miR-122 target site from 3′UTR of the VDR transcript, but not when this site was mutated. Moreover, miR-122 mimic significantly blocked paricalcitol-induced luciferase activity in 293T cells co-transfected with a VDRE-driven luciferase reporter, whereas miR-122 hairpin inhibitor enhanced paricalcitol’s activity to suppress PUMA and caspase 3 activation induced by LPS in HK-2 cells.

Conclusions
Collectively, these studies provide evidence that miR-122 directly targets VDR in renal tubular cells, which strongly suggest that miR-122 up-regulation in the kidney under LPS challenge contributes to kidney injury by down-regulating VDR expression.

He, J., Du, J., Yi, B., et al. MicroRNA-122 contributes to lipopolysaccharide-induced acute kidney injury via down-regulating the vitamin D receptor in the kidney. European Journal of Clinical Investigation. (2021) Full Text Here.

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The Eagle Bioscience’s Coronavirus COVID-19 IgM ELISA Assay Kit was recently highlighted in a publication about a review on current diagnostic techniques for COVID-19. This review includes techniques such as RT-PCR, ddPCR, LAMP, CRISPR, and immunoassay techniques. To learn more about this publication, read below:

Abstract

Introduction
SARS-Cov-2 first appeared in Wuhan, China, in December 2019 and spread all over the world soon after that. Given the infectious nature of SARS-CoV-2, fast and accurate diagnosis tools are important to detect the virus. In this review, we discuss the different diagnostic tests that are currently being implemented in laboratories and provide a description of various COVID-19 kits.

Areas Covered
We summarize molecular techniques that target the viral load, serological methods used for SARS-CoV-2 specific antibodies detection as well as newly developed faster assays for the detection of SARS-CoV-2 in various biological samples.

Expert Opinion
In the light of the widespread pandemic, the massive diagnosis of COVID-19, using various detection techniques, appears to be the most effective strategy for monitoring and containing its propagation.

Jaddaoui, IE., Allali, M., Raoui, S., et al. A review on current diagnostic techniques for COVID-19. Expert Review of Molecular Diagnostics. (2021) 21:2, 141-160.

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The Eagle Bioscience’s TNF-Alpha ELISA Assay Kit was recently utilized in a publication about the detection rate and genotyping of Cryptosporidium spp. and its relation to corporate TNF-Alpha in elderly Egyptians.

Abstract

Background
Elderly individuals are considered an at-risk population, susceptible to enteric infections; and Cryptosporidium spp. is an apicomplexan protozoan considered to be one of the most common protozoa causing diarrhea. Cryptosporidiosis causes elevation of many pro-inflammatory cytokines like tumor necrosis factor-alpha (TNF-α) which may play a role in pathogenesis of the disease.

Objectives
This study was designed for detection and genotyping of Cryptosporidium spp. in elderly patients and the relationship of infection with copro TNF-α. Diagnosis was by evaluation of permanent acid-fast cold Kinyoun’s (AF) staining, immunochromatographic detection (ICT), and ELISA in comparison to molecular diagnosis as gold standard diagnostic method.

Subjects and Methods
Stool samples were collected from 270 elderly patients aged above 60 years old attending outpatient clinics of Internal Medicine Hospital, Cairo University. Sample were examined microscopically by direct wet mount, and AF straining, and then subjected to ICT ELISA and nested PCR (nPCR) assays. Positive samples by nPCR were then subjected to Restriction fragment length polymorphism (RELP) to detect Cryptosporidium genotypes. Corpo-levels of TNF-Alpha were measured to asses their relationship with cryptosporidiosis.

Results
Cryptosporidiosis detection rates of 3.7%, 6.3%, 6.7%, 3.7% were determined by microscopic examination after AF staining, ICT, ELISA and nPCR, respectively. When RFLP was performed on nPCR positive samples, eight and two samples were assigned as genotype 1 and 2, respectively. Moreover, TNF-α was significantly correlated with cryptosporidiosis.

Conclusion
Conclusion: The elderly are highly vulnerable to cryptosporidiosis. Immunodiagnosis and molecular techniques are fundamental for the diagnosis of cryptosporidiosis. Cryptosporidiosis significantly affects copro TNF-α.

Amin, NM., Raafat, A., Morsy, SM. Detection rate and genotyping of Cryptosporidium spp. and its relation to copro TNF-α in elderly Egyptians attending outpatient clinics of Cairo University Hospitals. Parasitologists. (2021)14:77-85

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The Eagle Bioscience’s Anti-Infliximab ELISA Assay Kit was recently highlighted in a publication about factors that are associated with reduced infliximab exposure in the treatment of pediatric autoimmune disorders.

Abstract

Background
Inadequate systemic exposure to infliximab (IFX) is associated with treatment failure. This work evaluated factors associated with reduced IFX exposure in children with autoimmune disorders requiring IFX therapy.

Methods
In this single-center cross-sectional prospective study IFX trough concentrations and anti-drug antibodies (ADAs) were measured in serum from children diagnosed with inflammatory bowel disease (IBD) (n = 73), juvenile idiopathic arthritis (JIA) (n = 16), or uveitis (n = 8) receiving maintenance IFX infusions at an outpatient infusion clinic in a tertiary academic pediatric hospital. IFX concentrations in combination with population pharmacokinetic modeling were used to estimate IFX clearance. Patient demographic and clinical data were collected by chart review and evaluated for their relationship with IFX clearance.

Results
IFX trough concentrations ranged from 0 to > 40 μg/mL and were 3-fold lower in children with IBD compared to children with JIA (p = 0.0002) or uveitis (p = 0.001). Children with IBD were found to receive lower IFX doses with longer dosing intervals, resulting in dose intensities (mg/kg/day) that were 2-fold lower compared to children with JIA (p = 0.0002) or uveitis (p = 0.02). Use of population pharmacokinetic analysis to normalize for variation in dosing practices demonstrated that increased IFX clearance was associated with ADA positivity (p = 0.004), male gender (p = 0.02), elevated erythrocyte sedimentation rate (ESR) (p = 0.02), elevated c-reactive protein (CRP) (p = 0.001), reduced serum albumin concentrations (p = 0.0005), and increased disease activity in JIA (p = 0.009) and IBD (p ≤ 0.08). No significant relationship between diagnosis and underlying differences in IFX clearance was observed. Multivariable analysis by covariate population pharmacokinetic modeling confirmed increased IFX clearance to be associated with anti-IFX antibody positivity, increased ESR, and reduced serum albumin concentrations.

Conclusions
Enhanced IFX clearance is associated with immunogenicity and inflammatory burden across autoimmune disorders. Higher systemic IFX exposures observed in children with rheumatologic disorders are driven primarily by provider drug dose and interval selection, rather than differences in IFX pharmacokinetics across diagnoses. Despite maintenance IFX dosing at or above the standard recommended range for IBD (i.e., 5 mg/kg every 8 weeks), the dosing intensity used in the treatment of IBD is notably lower than dosing intensities used to treat JIA and uveitis, and may place some children with IBD at risk for suboptimal maintenance IFX exposures necessary for treatment response.

Funk, RS., Shakhnovich, V., Cho, YK., et al. Factors associated with reduced infliximab exposure in the treatment of pediatric autoimmun disorders: a cross-sectional prospective convenience sampling study. Pediatric Rheumatology. (2021) 19:62

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The Eagle Bioscience’s Calprotectin ELISA Assay Kit was highlighted in a recent study! In this publication scientists studied the dose effect of bovine lactoferrin (bLF) fortification on diarrhea and respiratory tract infections in weaned infants with anemia.

Abstract

Objective
The aim of this study was to explore the dose effect of bovine lactoferrin (bLF) fortification on the morbidity of diarrhea and respiratory tract infections in weaned infants with anemia.

Methods
A total of 108 infants with anemia, who were exclusively breast fed at 4 to 6 months and weaned and formula fed at 6 to 9 months, were recruited. The eligible infants were randomly assigned to fortified group 0 (FG0), fortified group 1 (FG1), or fortified group 2 (FG2) and were given formula fortified with 0 mg/100 g, 38 mg/100 g, and 76 mg/100 g of bLF, respectively, for 3 mo. The morbidity of diarrhea and respiratory tract infections (RTIs), the duration of respiratory and diarrhea-related illnesses, and the levels of fecal human beta-defensin 2 (HBD-2), cathelicidin LL-37 (LL-37), secretory IgA (sIgA), butyrate, and calprotectin were assessed.

Results
After the exclusion of 12 dropouts, the primary outcome measures, including episodes and duration of diarrhea and RTIs during the intervention, were obtained from 96 infants (35, 33, and 28 in FG0, FG1, and FG2, respectively). Compared with infants in FG0, there was a lower morbidity of rhinorrhea, wheezing, and skin rash among infants in FG1 (P < 0.05) and a lower morbidity of respiratory-related illness and wheezing among infants in FG2 (P < 0.05). Furthermore, a lower morbidity of diarrhea-related illness, diarrhea, vomiting, and nausea was observed among infants in FG2 than those in the other two groups (P < 0.05). In addition, the FG1 infants had a lower morbidity of vomiting and nausea than the FG0 infants (P < 0.05). The HBD-2, LL-37, sIgA, and calprotectin levels were significantly higher whereas the butyrate level was significantly lower in the FG2 infants than in infants in the other two groups after 3 mo of intervention (P < 0.05).

Conclusions
The bLF-fortified formula was effective in reducing the morbidity of diarrhea and RTIs in infants with anemia, with the 76 mg/100 g bLF-fortified formula exhibiting a stronger effect. The bLF fortification could be a new strategy for the prevention of diarrhea and RTIs in infants with anemia.

Chen, K., Jin, S., Chen, H.,et al. Dose effect of bovine lactoferrin fortification on diarrhea and respiratory tract infections in weaned infants with anemia: A randomized, controlled trial. Nutrition. (2021)90:111288

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The Eagle Bioscience’s ACTH ELISA Assay Kit was recently highlighted in a publication about the cortisol and adrenocorticotrophic hormone (ACTH) in infants after receiving corticosteroids following cataract surgery.

Abstract

Purpose
Cushingoid features are occasionally encountered in infants after pediatric cataract surgery. The aim of this study is to evaluate whether the use of topical glucocorticoids (GCs) following congenital cataract surgery can result in endogenous adrenal suppression and/or systemic side effects similar to those seen with systemic steroids.

Methods
A prospective study was performed on 20 infants with bilateral congenital cataract. All infants received a single subconjunctival betamethasone injection of 1 mg at the end of surgery in addition to topical dexamethasone eye drops 1 mg/ml for 6 weeks. All infants had anthropometric measurements and blood pressure measurements, serum cortisol, and ACTH level measurements before surgery and 2 months after. In addition, the total administered glucocorticoid adjusted per weight was calculated.

Results
The mean age of the infants was 4.93 ± 2.58 months. Thirteen were males (65%). The total administered glucocorticoid dose was 18.7 mg and the mean cumulative dexamethasone equivalent dose administered was 2.75 ± 1.31 mg/kg. There was a statistically significant increase in the adjusted weight percentile for age (P = 0.009). Both the systolic and diastolic blood pressure were significantly elevated (P = 0.005 and P = 0.025 respectively). There was a statistically significant reduction in both the morning and afternoon serum ACTH levels (P = 0.023 and P = 0.014). The reduction in serum cortisol levels was statistically non-significant.

Conclusions
Topical steroids following pediatric cataract surgery can result in both subclinical and clinical changes in the hypothalamic–pituitary–adrenal axis that can be easily overlooked and need careful attention and follow-up.

Aly, A., Gouda, J., Awadein, A. et al. Serum cortisol and adrenocorticotrophic hormone (ACTH) in infants receiving topical and subconjunctival corticosteroids following cataract surgery. Graefes Arch Clin Exp Ophthalmol (2021). https://doi.org/10.1007/s00417-021-05221-0

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The Eagle Biosciences’ Dopamine ELISA Assay Kit was used recently in a Parkinson study. This study aimed to explore the neuroprotective effect that tiron could have against MPTP-induced Parkinsonism. Read more about this study below.

Abstract

Parkinsonism is a neurodegenerative disease that is common all over the world. This study aimed at exploring the neuroprotective effect of tiron against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinsonism. MPTP (30 mg/kg, intraperitoneally [ip]) was injected in mice daily for 5 consecutive days. Mice were treated with tiron (140 and 280 mg/kg, ip) or levodopa (8.4 mg/kg, orally) for 10 consecutive days starting 5 days before MPTP injection. At the end of the experiment, behavioral tests were conducted to assess the neuroprotective effect of tiron. Moreover, oxidative stress was assessed via measuring antioxidant enzyme, such as catalase, and lipid peroxidation was evaluated as malondialdehyde. Neuronal damage was also detected by histopathological examination and via estimating hippocampal levels of dopamine, γ-aminobutyric acid, and nuclear factor erythroid-derived 2-like 2. In addition, the expression of Kelch-like ECH-associated protein 1 and heme oxygenase-1 was assessed by immunohistochemistry. Compared with the blank control group and the positive control group, the inhibitory effect of tiron on MPTP-induced neurodegenerative injury was statistically significant.

Mohamed SA, El-Kashef DH, Nader MA. Tiron Alleviates MPTP-Inceded Parkisonism in Mice Via Activation of Keap-1/Nrf2 Pathway. J Biochem Mol Toxicol. 2020;35:e22685.

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