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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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