Author: Eagle Biosciences

Clostridium Difficile Toxin AB Qualitative ELISA Assay Kit was utilized in a recent study! Researchers demonstrated that treatment with the microbial metabolite urolithin A (UroA) attenuates Clostridioides difficile infection (CDI) induced adverse effects on the colon epithelium in a preclinical model of CDI-induced colitis. Check out the abstract and full text below.

Abstract

Clostridioides difficile is a Gram-positive, anaerobic, spore-forming bacterium responsible for antibiotic-associated pseudomembranous colitis. Clostridioides difficile infection (CDI) symptoms can range from diarrhea to life-threatening colon damage. Toxins produced by C. difficile (TcdA and TcdB) cause intestinal epithelial injury and lead to severe gut barrier dysfunction, stem cell damage, and impaired regeneration of the gut epithelium. Current treatment options for intestinal repair are limited. In this study, we demonstrate that treatment with the microbial metabolite urolithin A (UroA) attenuates CDI-induced adverse effects on the colon epithelium in a preclinical model of CDI-induced colitis. Moreover, our analysis suggests that UroA treatment protects against C. difficile-induced inflammation, disruption of gut barrier integrity, and intestinal tight junction proteins in the colon of CDI mice. Importantly, UroA treatment significantly reduced the expression and release of toxins from C. difficile without inducing bacterial cell death. These results indicate the direct regulatory effects of UroA on bacterial gene regulation. Overall, our findings reveal a novel aspect of UroA activity, as it appears to act at both the bacterial and host levels to protect against CDI-induced colitis pathogenesis. This research sheds light on a promising avenue for the development of novel treatments for C. difficile infection.

Ghosh S, Erickson D, Chua MJ, Collins J, Jala VR. 2024. The microbial metabolite urolithin A reduces Clostridioides difficile toxin expression and toxin-induced epithelial damage. mSystems 9:e01255-23. https://doi.org/10.1128/msystems.01255-23

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Check out this recent publication that utilized our Calprotectin ELISA Assay Kit! This study examined whether patient-derived materials could predict individual clinical responsiveness to the Janus kinase (JAK) inhibitory, tofacitinib, prior to treatment initiation. Find the abstract and full text below.

Abstract

Background & Aims: Despite increasing therapeutic options in the treatment of ulcerative colitis (UC), achieving disease remission remains a major clinical challenge. Nonresponse to therapy is common and clinicians have little guidance in selecting the optimal therapy for an individual patient. This study examined whether patient-derived materials could predict individual clinical responsiveness to the Janus kinase (JAK) inhibitor, tofacitinib, prior to treatment initiation.

Method: In 48 patients with UC initiating tofacitinib, we longitudinally collected clinical covariates, stool, and colonic biopsies to analyze the microbiota, transcriptome, and exome variations associated with clinical responsiveness at week 24. We established patient-derived organoids (n = 23) to determine how their viability upon stimulation with proinflammatory cytokines in the presence of tofacitinib related to drug responsiveness in patients. We performed additional biochemical analyses of organoids and primary tissues to identify the mechanism underlying differential tofacitinib sensitivity.

Results: The composition of the gut microbiota, rectal transcriptome, inflammatory biomarkers, and exome variations were indistinguishable among UC patients prior to tofacitinib treatment. However, a subset of patient-derived organoids displayed reduced sensitivity to tofacitinib as determined by the ability of the drug to inhibit STAT1 phosphorylation and loss of viability upon cytokine stimulation. Remarkably, sensitivity of organoids to tofacitinib predicted individual clinical patient responsiveness. Reduced responsiveness to tofacitinib was associated with decreased levels of the cationic transporter MATE1, which mediates tofacitinib uptake.

Conclusions: Patient-derived intestinal organoids predict and identify mechanisms of individual tofacitinib responsiveness in UC. Specifically, MATE1 expression predicted clinical response to tofacitinib.

Jang KK, Ercelen D, Cen Feng JYC, et al. Tofacitinib uptake by patient-derived intestinal organoids predicts individual clinical responsiveness. bioRxiv [Preprint]. 2024 Mar 6:2024. doi: 10.1101/2024.03.02.583137

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Check out this recent article that utilized our H. pylori Qualitative ELISA! This study aimed to determine the association of H. Pylori infection among acne vulgaris (AV) patients and correlate it with disease activity. Find the abstract and full text below.

Abstract

Background: Helicobacter pylori (H. pylori) is a gastric Gram-negative, spiral-shaped microaerophilic pathogen. H. pylori may play a potential pathogenic role in extra-intestinal diseases such as hepatobiliary, respiratory, and dermatological disorders. The latter included chronic urticaria, psoriasis and rosacea. The first report in literature on the relationship between H. pylori and acne vulgaris (AV), found association between severe AV and H. pylori infection. There are very limited data in AV patients addressing the impact of H. pylori infection on various severities. In this context, the aim of the present work was to determine the association of H. Pylori infection among AV patients and correlate it with the disease severity.

Methods: This case-control study included 45 Patients with AV and 45 age and sex matched healthy volunteers as a control group. H. pylori antigen in stool and serum H. pylori antibody IgG using commercially available ELISA kits was tested in all included subjects.

Results: The percentage of participants with a positive H. pylori antigen in stool and positive H. pylori antibody in serum in the whole study population was 35/90 (38. 9%) and 41/90 (45. 6%). On comparing between the percentagesof positive H. pylori antigen in stool and positive H. pylori antibody in serum between the patients with AV and healthy controls, a highly statistically significant difference was found between the two groups (P<0.001, P=0.006). On comparing between the percentages of positive H. pylori antigen in stool and positive H. pylori antibody in serum in the patients with different grades of acne severity and healthy controls, the rate of positive H. pylori antigen in stool and positive H. pylori Ab in serum was significantly associated with severity of acne comparing with healthy controls (p<0. 001).

Conclusion: The rate of H. pylori infection in patients with AV is high so it may influence the pathogenesis of this skin disease. Patients with severe AV had higher rates of H. pylori antigen in stool and H. pylori antibody in serum as compared to the patients with mild AV and healthy controls.

ahmed afify, Hanan Mohamed Ahmed Saleh, Abeer Farrag et al. Helicobacter pylori and acne vulgaris: is there a relationship?, 08 January 2024, PREPRINT (Version 1) available at Research Square https://doi.org/10.21203/rs.3.rs-3835029/v1

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Check out this recent study that utilized our Noradrenaline (Norepinephrine) Sensitive ELISA! The researchers aimed to develop a method to innervate human pluripotent stem cell (hPSC)-based 3D organoids, particularly focusing on the autonomic sympathetic nervous system’s regulation of the heart. Check out the abstract and full text below.

Abstract

The technology of human pluripotent stem cell (hPSC)-based 3D organoid/assembloid cultures has become a powerful tool for the study of human embryonic development, disease modeling and drug discovery in recent years. The autonomic sympathetic nervous system innervates and regulates almost all organs in the body, including the heart. Yet, most reported organoids to date are not innervated, thus lacking proper neural regulation, and hindering reciprocal tissue maturation. Here, we developed a simple and versatile sympathetic neuron (symN)-innervated cardiac assembloid without the need for bioengineering. Our human sympathetic cardiac assembloids (hSCAs) showed mature muscle structures, atrial to ventricular patterning, and spontaneous beating. hSCA-innervating symNs displayed neurotransmitter synthesis and functional regulation of the cardiac beating rate, which could be manipulated pharmacologically or optogenetically. We modeled symN-mediated cardiac development and myocardial infarction. This hSCAs provides a tool for future neurocardiotoxicity screening approaches and is highly versatile and modular, where the types of neuron (symN or parasympathetic or sensory neuron) and organoid (heart, lung, kidney) to be innervated may be interchanged.

Nadja Zeltner, Hsueh-Fu Wu, Kenyi Saito-Diaz et al. A modular platform to generate functional sympathetic neuron-innervated heart assembloids, 21 March 2024, PREPRINT (Version 1) available at Research Square https://doi.org/10.21203/rs.3.rs-3894397/v1

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The StressMarq Biosciences, Inc. Alpha B Crystallin ELISA was utilized in a recent publication! The study aimed to investigate the potential peripheral myelin protein 2 (P2) and Alpha B-crystallin (aBC) as predictive biomarkers in Guillain-Barré syndrome (GBS). Check out the abstract and full text below!

Abstract

Objectives: The study aimed to investigate the potential of peripheral myelin protein 2 (P2) and Alpha B-crystallin (αBC) as predictive biomarkers in Guillain-Barré syndrome (GBS). Given the unpredictability of GBS prognosis, the need for specific and reliable biomarkers for disease development and intensity assessment is crucial.

Material and Methods: A prospective observational study was conducted on a cohort of 220 individuals diagnosed with GBS at a tertiary general hospital in South India. P2 and αBC levels in cerebrospinal fluid (CSF) were quantified using ELISA assay kits. The study spanned from March 2021 to April 2023, with participants aged 18–60 years. The study protocol adhered to ethical standards, and the Brighton criteria were employed for GBS diagnosis. CSF samples were collected at admission and two weeks post-onset. Data analysis utilised SPSS, and statistical significance was set at p < 0.05.

Results: Upon admission, mean P2 levels were 2.2 ± 0.5 ng/mL, and αBC levels were 9.8 ± 2.3 ng/mL. After two weeks, P2 increased to 4.8 ± 0.8 ng/mL, and αBC increased to 15.1 ± 2.3 ng/mL. A positive correlation was observed between the rise in P2 and αBC levels and enhanced muscle strength at 4 weeks and 6 months.

Conclusion: The study suggests a significant increase in P2 and αBC levels in GBS patients, correlating with improved muscle strength. P2 and αBC ratios in CSF between the second and first weeks may serve as prognostic markers for GBS. Limitations include a small sample size and the absence of a control group, necessitating caution in generalizability.

Amalakanti S, Arepalli KVR, Jillella JP. Utility of cerebrospinal fluid protein biomarkers in predicting the outcome of Guillain-Barre syndrome. South Asian J Health Sci. 2024;1:27–30. doi: 10.25259/SAJHS_18_2023

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The Biomedica Sclerostin ELISA Assay Kit was utilized in a recent publication! This study assessed the associations between serum and bone sclerostin levels and biomarkers of bone turnover and bone histomorphometry. Check out the abstract and full text below.

Abstract

The pathophysiology of chronic kidney disease-mineral and bone disorder (CKD-MBD) is not well understood. Specific factors secreted by osteocytes are elevated in the serum of adults and pediatric patients with CKD-MBD, including FGF-23 and sclerostin, a known inhibitor of the Wnt signaling pathway. The molecular mechanisms that promote bone disease during the progression of CKD are incompletely understood. In this study, we performed a cross-sectional analysis of 87 pediatric patients with pre-dialysis CKD and post-dialysis (CKD 5D). We assessed the associations between serum and bone sclerostin levels and biomarkers of bone turnover and bone histomorphometry. We report that serum sclerostin levels were elevated in both early and late CKD. Higher circulating and bone sclerostin levels were associated with histomorphometric parameters of bone turnover and mineralization. Immunofluorescence analyses of bone biopsies evaluated osteocyte staining of antibodies towards the canonical Wnt target, β-catenin, in the phosphorylated (inhibited) or unphosphorylated (active) forms. Bone sclerostin was found to be colocalized with phosphorylated β-catenin, which suggests that Wnt signaling was inhibited. In patients with low serum sclerostin levels, increased unphosphorylated “active” β-catenin staining was observed in osteocytes. These data provide new mechanistic insight into the pathogenesis of CKD-MBD and suggest that sclerostin may offer a potential biomarker or therapeutic target in pediatric renal osteodystrophy.

Laster M. et al. Sclerostin, Osteocytes, and Wnt Signaling in Pediatric Renal Osteodystrophy. Nutrients. 2023 Sep 25;15(19):4127. doi: 10.3390/nu15194127

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Our Eagle Bioscience’s NGAL Stool ELISA Assay Kit was utilized in a recent study! This study evaluated fecal inflammatory biomarkers to identify bacterial diarrhea episodes. Check out the abstract and full text below.

Abstract

Background: The measurement of fecal inflammatory biomarkers among individuals presenting to care with diarrhea could improve the identification of bacterial diarrheal episodes that would benefit from antibiotic therapy. We reviewed prior literature in this area and describe our proposed methods to evaluate 4 biomarkers in the Enterics for Global Health (EFGH) Shigella surveillance study.

Methods: We systematically reviewed studies since 1970 from PubMed and Embase that assessed the diagnostic characteristics of inflammatory biomarkers to identify bacterial diarrhea episodes. We extracted sensitivity and specificity and summarized the evidence by biomarker and diarrhea etiology. In EFGH, we propose using commercial enzyme-linked immunosorbent assays to test for myeloperoxidase, calprotectin, lipocalin-2, and hemoglobin in stored whole stool samples collected within 24 hours of enrollment from participants in the Bangladesh, Kenya, Malawi, Pakistan, Peru, and The Gambia sites. We will develop clinical prediction scores that incorporate the inflammatory biomarkers and evaluate their ability to identify Shigella and other bacterial etiologies of diarrhea as determined by quantitative polymerase chain reaction (qPCR).

Results: Forty-nine studies that assessed fecal leukocytes (n = 39), red blood cells (n = 26), lactoferrin (n = 13), calprotectin (n = 8), and myeloperoxidase (n = 1) were included in the systematic review. Sensitivities were high for identifying Shigella, moderate for identifying any bacteria, and comparable across biomarkers. Specificities varied depending on the outcomes assessed. Prior studies were generally small, identified red and white blood cells by microscopy, and used insensitive gold standard diagnostics, such as conventional bacteriological culture for pathogen detection.

Conclusions: Our evaluation of inflammatory biomarkers to distinguish diarrhea etiologies as determined by qPCR will provide an important addition to the prior literature, which was likely biased by the limited sensitivity of the gold standard diagnostics used. We will determine whether point-of-care biomarker tests could be a viable strategy to inform treatment decision making and increase appropriate targeting of antibiotic treatment to bacterial diarrhea episodes

Babb, C et al. Evaluation of Fecal Inflammatory Biomarkers to Identify Bacterial Diarrhea Episodes: Systematic Review and Protocol for the Enterics for Global Health Shigella Surveillance Study, Open Forum Infectious Diseases, Volume 11, Issue Supplement_1, March 2024, Pages S65–S75, https://doi.org/10.1093/ofid/ofad652

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Check out this recent study that highlighted our Serotonin Sensitive ELISA Kit! This study tested whether the perturbed prenatal and postnatal dietary exposures affect the developing offspring predisposing to neurobehavioral disorders in the adult. Check out the abstract and full text below.

Abstract

Introduction: Serotonin (5-HT) is critical for neurodevelopment and the serotonin transporter (SERT) modulates serotonin levels. Perturbed prenatal and postnatal dietary exposures affect the developing offspring predisposing to neurobehavioral disorders in the adult. We hypothesized that the postnatal brain 5-HT-SERT imbalance associated with gut dysbiosis forms the contributing gut-brain axis dependent mechanism responsible for such ultimate phenotypes.

Methods: Employing maternal diet restricted (IUGR, n=8) and high fat+high fructose (HFhf, n=6) dietary modifications, rodent brain serotonin was assessed temporally by ELISA and SERT by quantitative Western blot analysis. Simultaneously, colonic microbiome studies were performed.

Results: At early postnatal (P) day 2 no changes in the IUGR, but a ~24% reduction in serotonin (p = 0.00005) in the HFhf group occurred, particularly in the males (p = 0.000007) revealing a male versus female difference (p = 0.006). No such changes in SERT concentrations emerged. At late P21 the IUGR group reared on HFhf (IUGR/HFhf, (n = 4) diet revealed increased serotonin by ~53% in males (p = 0.0001) and 36% in females (p = 0.023). While only females demonstrated a ~40% decrease in serotonin (p = 0.010), the males only trended lower without a significant change within the HFhf group (p = 0.146). SERT on the other hand was no different in HFhf or IUGR/RC, with only the female IUGR/HFhf revealing a 28% decrease (p = 0.036). In colonic microbiome studies, serotonin-producing Bacteriodes increased with decreased Lactobacillus at P2, while the serotonin-producing Streptococcus species increased in IUGR/HFhf at P21. Sex-specific changes emerged in association with brain serotonin or SERT in the case of Alistipase, Anaeroplasma, Blautia, Doria, Lactococcus, Proteus, and Roseburia genera.

Discussion: We conclude that an imbalanced 5-HT-SERT axis during postnatal brain development is sex-specific and induced by maternal dietary modifications related to postnatal gut dysbiosis. We speculate that these early changes albeit transient may permanently alter critical neural maturational processes affecting circuitry formation, thereby perturbing the neuropsychiatric equipoise.

Ye X, et al. (2024) Brain serotonin and serotonin transporter expression in male and female postnatal rat offspring in response to perturbed early life dietary exposures. Front. Neurosci. 18:1363094. doi: 10.3389/fnins.2024.1363094

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