#MYD88 Mutation Detection Kit

[blackbiobiotech]

Understanding ROS1 and MYD88 Mutations: Key Diagnostic Tools for Oncologists

Cancer research and treatment are rapidly evolving, with molecular diagnostics playing a pivotal role in shaping personalized treatment plans. With advances in genetic testing, clinicians can identify specific mutations that drive the development of various cancers, allowing for more targeted therapies. Among the most important genetic markers in cancer diagnostics are ROS1 and MYD88 mutations. These mutations are linked to specific cancers, including non-small cell lung cancer (NSCLC) and certain lymphomas. 3B BlackBio Biotech offers cutting-edge solutions for detecting these mutations, including the ROS1 PCR Kit and MYD88 Mutation Detection Kit. These kits help clinicians make informed decisions based on precise genetic data.

ROS1 PCR Kit: Unveiling Key Insights for Lung Cancer Treatment

ROS1 gene fusions are known to occur in a subset of non-small cell lung cancers (NSCLC). ROS1 mutations are associated with a high potential for targeted therapy, as they make tumors sensitive to certain types of tyrosine kinase inhibitors (TKIs). However, accurately detecting ROS1 gene rearrangements is critical for determining the suitability of these therapies.

3B BlackBio Biotech’s ROS1 Gene Fusions Detection Kit offers a highly sensitive and reliable method for identifying ROS1 mutations. By detecting gene fusions, this kit provides valuable diagnostic information that can directly influence treatment choices. ROS1 mutations are relatively rare, but their detection is essential for selecting the most effective therapy. This PCR-based kit uses advanced techniques to ensure accurate results, making it an essential tool for oncologists looking to personalize cancer treatments.

The ROS1 mutations kit is particularly important for NSCLC patients who may benefit from therapies like crizotinib, an FDA-approved drug that targets ROS1 gene rearrangements. The ability to identify these mutations early on ensures that patients receive the best possible care, improving their prognosis and quality of life.

MYD88 PCR Kit: A Key Player in Lymphoma Diagnostics

MYD88 mutations are commonly found in various types of lymphoma, particularly in Waldenström macroglobulinemia (WM), a rare and slow-growing cancer of the blood. MYD88 mutations are crucial for diagnosing and predicting the course of the disease, as well as determining the most effective treatment options. The detection of MYD88 mutations allows for a more precise diagnosis and a tailored approach to treatment.

The MYD88 Mutation Detection Kit provides a powerful tool for identifying MYD88 mutations in patients with suspected lymphoma. This PCR-based test detects mutations in the MYD88 gene, which are linked to abnormal activation of the NF-κB pathway—a key factor in the development of lymphoma. Accurate detection of MYD88 mutations helps clinicians understand the molecular profile of the tumor, allowing them to choose the most appropriate treatment for the patient.

The MYD88 PCR kit is not only useful for diagnosing Waldenström macroglobulinemia but also for assessing other lymphoma subtypes. With its high sensitivity and precision, this kit plays a crucial role in the clinical management of lymphoma patients, improving diagnostic accuracy and treatment outcomes.

The Role of 3B BlackBio Biotech in Advancing Cancer Diagnostics

3B BlackBio Biotech has been at the forefront of providing high-quality diagnostic kits for genetic mutations associated with various cancers. Their ROS1 PCR Kit and MYD88 Mutation Detection Kit are prime examples of the company’s commitment to enhancing precision medicine. These kits enable clinicians to detect key mutations, ensuring that patients receive the most effective treatments based on their genetic profiles.

By offering advanced tools like the ROS1 Gene Fusions Detection Kit and the MYD88 Mutation Detection Kit, 3B BlackBio Biotech is contributing significantly to the personalization of cancer care. These diagnostic solutions help oncologists make data-driven decisions, ultimately improving patient outcomes and advancing the fight against cancer.

For more information on 3B BlackBio Biotech's advanced PCR kits, visit at https://3bblackbio.com/.

[lupine-publishers-lojpcr]

Lupine Publishers | Effects of Kav001 (A Kavain Analogue) on LPS-Induced Cytokines and Neutrophil Infiltration Lupine Publishers | LOJ Pharmacology & Clinical Research

Abstract

Background: TNF-α, a cytokine, is known to be involved in LPS-induced inflammation. Our recent data indicate that treatment of Kavain in mouse or human cells significantly suppresses TNF-α production in response to LPS. In order to completely identify one Kavain analogue (Kav001) reported in our previous data, we further analyzed its biological function in this paper.

Methods and Results: Commercial Kavain or Kav001 as inhibitor was used. ELISA was performed for detection of TNF-α, IL-1β, IL-6, IL-4, or Caspase 1 after treatment of THP-1 cells with Kav001. Wild type mice (C57BL/6) were used for neutrophil infiltration assay. A kinase/protein array was performed to analyze if Kav001 affects any kinases/protein in response to LPS. Conclusions: It is clear that Kav001 not only inhibits TNF-α, but also IL-1β, IL-6, caspase 1 and neutrophil infiltration in response to LPS compared to control.

Introduction

Kavain is known as a treatment for certain diseases [1-3]. Amar’s group recently reported that, in vivo, Kavain mediated alveolar bone loss and inflammatory infiltrate in a mnouse model [4]. Kavain is found to affect TNF-α production through regulation of NF-kB [5] and to be inhibited by ERK2 dephosphorylation in vitro or in vivo [6]. Our previous data indicate that Kavain inhibits P. gingivalis/LPS-induced inflammation/inflammatory disease [7,8]. Recently, we found that Kavain grants small amounts of toxicity to mammalian cells, most likely due to its insoluble chemical structure with hydrophobicity. To address this issue, we designed and synthesized compounds with the basic chemical structure of Kavain with a different number of hydrophilic bonds. One soluble compound (named Kav001) from the Kavain analogue was selected and further confirmed to have a stronger biological function than Kavain by in vitro/in vivo experiments such as CAIA mouse models or endotoxic shock [9]. In this paper, we further found that Kav001 not only inhibits TNF-α, but also IL-1β, IL-6, caspase 1 and neutrophil infiltration. However, this phenomenon cannot be observed when macrophages are treated with LPS plus Kavain. We believe that Kav001 may mediate a novel link between Kav001 and LPS-induced inflammation and may be used as a key inhibitor to LPS-induced inflammation/inflammatory disease.

Materials and Methods

Animals and cells

Wild type mice (8-10 week-old C57BL/6, Jackson Labs) were maintained under strict specific pathogen-free (SPF) conditions. All protocols were approved by the Boston University Institutional Animal Care and Use Committee and were performed in compliance with the relevant animal care and use laws and institutional guidelines. THP-1 cells (TIB-202, ATCC) were cultured in a RPMI- 1640 media (Cat#: 11875–093, Life Technologies, NY) with 10% FBS at 37 °C in a 5% CO2 atmosphere.

Compounds

Commercial Kavain (Cat#59780, Sigma‐Aldrich Co., Natick, MA) and custom synthesized analogue Kav001 (Lot#KB180125, KareBay Biochem, Inc.) were obtained. The analysis of each compound shows that ≥95% purity. The stock concentration of each compound was prepared with 10μg/μl. The compound was diluted to 100μg/ml in 25% DMSO for Kavain or in water for Kav001 just before the experiment.

Elisa

The treated media from THP-1 cells were subjected to ELISA for TNF-α detection with the kit (Cat#: ab181421, Abcam), IL-1β detection with the kit (Cat#: ab100562, Abcam), IL-6 detection with the kit (Cat#: KHC0061, Life technologies), IL-4 detection with the kit (Cat#: BMS225-2, Thermo Fisher), or Caspase 1 (Cat#: ab219633, Abcam). ELISA immunoreactivity was measured by a reader (Model 680, Bio-Rad). The data were graphed.

Neutrophil Infiltration Assay

Mice (n=3) were untreated or treated by i.p. injection of E.coli LPS (1mg/kg body weight) and/or oral gavage of Kav001 (30mg/ kg) for 24hrs. Skin tissues from each group were taken and fixed in 4% paraformaldehyde, then embedded in O.C.T. compound [10,11]. Skin sections (5μm) were made and stained with H&E (hematoxylin and eosin). Results were analyzed and imaged.

Protein Array

1 × 105 THP-1 cells were treated with 100μg/ml of E. coli LPS (Sigma) or plus Kav001 (0.1mg/ml) for 16 hrs at 37 °C. The medium from each test was centrifuged at 1,500 × g and then treated with a specific kit for human protein cytokine array (RayBiotech, Norcross, GA) based on the manufacturer’s instructions. The data were analyzed using a Verrado Imaging System (Bio-Rad) and graphed.

Statistical Analysis

All tests were done in triplicate and statistical analyses were done with the SAS software package. All data were normally distributed. For multiple mean comparisons, we conducted ANOVA analysis by the Student’s t-test for single mean comparison. P values less than 0.05 were considered significant.

Results

In order to examine whether Kav001 also inhibits other cytokines besides TNF-α, we made a relevant experiment via ELISA. As shown in Figure 1A-1C, co-treatment of LPS plus Kav001 (A-C, #4) significantly reduces LPS-induced TNF-α, IL-1β, or IL-6 compared to the positive controls (A-C, #2). This phenomenon for detection of TNF-α was also observed while co-treatment of LPS plus Kavain but was not detectable for IL-1β or IL-6 production (1B&C, #3) As shown in Figure 1D,1B. Additionally, neither Kavain nor Kav001 was found to affect IL-4. Because our preliminary data indicate that Kav001 affects some cytokines in response to LPS, we were therefore interested in whether Kav001 is involved in other stimuli-mediated cytokines. As shown in Figure 1E & 1G, Kav001 strongly reduced all three stimuli-induced cytokine productions, TNF-α (E, No.4), IL-1β (F, No.4), and IL-6 (G, No.4) compared to their positive control (No.2 of E, F, or G). This phenomenon for reduction of IL-1β (F, No.3) or IL-6 (G, No.3) was not observed with co-treatment of stimuli plus Kavain. Since it is known that caspase 1 expression is activated with TNF-α, IL-1 beta, or IL-6 [12-14], we wanted to further test whether Kav001 inhibits caspase 1 caused by TNF-α, IL-1 beta, or IL-6. As shown in Figure 1F & 1H, Kav001 (H, No.6) reduced LPS-induced caspase 1 compared to the positive control (H, No.2).

As a control, Kavain did not reduce LPS-induced caspase 1, but instead increased its value (H, No.5). In order to analyze whether the treatment of Kav001 affects any kinases/protein upstream of caspase 1 in response to LPS, a kinase/protein array was performed. As shown in Figure 2B & 2C Kav001 significantly reduced LPSinduced kinases such as Akt (Protein kinase B) at 17%, ATM (Ataxia telangiectasia mutated) at 13%, caspase 3 at 11.5%, caspase 7 at 8.6%, CHK1 (Checkpoint kinase 1) at 26.4%, CHK2 (Checkpoint kinase 2) at 15.3%, and ERK1/2 at 12.7%/14.8%, but does not affect p27, p53, or TAK1 (Transforming growth factor beta-activated kinase 1 (Figure 2A & 2D). As it is known that subcutaneous injection of LPS in mice induces neutrophil infiltration of tissues [15,16], we were therefore interested in whether Kav001 treatment is able to suppress LPS-induced neutrophil infiltration. As shown in Figure 3A & 3D, indeed, subcutaneous treatment of 30mg/kg of Kav001 significantly reduced LPS-induced Neutrophil infiltration of tissues in mouse (DI,II, III) compared to the positive control (CI,II, III).

Figure 2:Kinase/Protein array. (A) The stamped identification number on the array. (B) THP-1 cells were untreated or treated with LPS alone or LPS+ Kav001 over night. The conditioned medium from each test group was assessed by protein assay and quantified by a Versace Imaging System. (C) Analysis of the intensity on the array filter. The cell intensity treated by LPS was assigned to 100% as base value and compared to the others. There ratio was calculated in this table.

Discussion

Kavain was shown to affect TNF-α transcriptional regulation [6- 8]. Compared with Kavain, Kav001 not only inhibits LPS-induced TNF-α, , but also IL-1β, IL-6, caspase 1 or Neutrophil infiltration. DMSO is known to be widely used as reagent solvent but sometimes causes a little toxicity to mammalian cells [17]. It is known that Kavain is dissolved in the higher concentration of DMSO (≥25%) while Kav001 is completely dissolved in water. This water-soluble property may be the reason why Kav001 prevent DMSO-mediated toxicity to cells with a longer period of efficacy. Our most recent data support this potential hypothesis [9]. Additionally, neutrophils are the innate immune response and thus, neutrophilic infiltration of tissues is involved in many inflammatory diseases [18,19]. In order to inhibit the neutrophilic infiltration from tissue injury (I/R injury, brain injury, ethanol feeding-induced liver injury), several inhibitors such as MgIG (Magnesium isoglycyrrhizinate), Clenbuterol, Phillyrin, or PAI-1 (Plasminogen activator inhibitor-1) have been widely studied [20,21]. Researcher groups have also suggested that these inhibitors suppress infiltration of neutrophils via regulation of factors such as MyD88 or p38 MAPK [22,23]. However, these studies of certain inhibitors leave several questions, such as whether there is a common key point between their pathways. Our data here also show that the treatment of Kav001 significantly reduces LPS-induced neutrophilic infiltration. Thus, Kav001 may be used for suppression of neutrophil infiltration induced by other stimuli or injuries. Since TNF-α, IL-1β, or IL-6 was found here to be inhibited by Kav001, we assume that Kav001 may have treatment for inflammatory diseases caused by these cytokines (TNF-α, IL-1β, or IL-6). We will try to address this hypothesis for future research.

https://lupinepublishers.com/pharmacology-clinical-research-journal/pdf/LOJPCR.MS.ID.000122.pdf

https://lupinepublishers.com/pharmacology-clinical-research-journal/fulltext/effects-of-kav001-(a-kavain-analogue)-on-lps-induced-cytokines-and-neutrophil-infiltration.ID.000122.php

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Advanced Insights into ROS1 and MYD88 Mutation Detection Kits for Precision Oncology

In the evolving field of molecular diagnostics, early and accurate detection of genetic mutations is pivotal in determining targeted treatments for various cancers. The ROS1 PCR Kit and MYD88 Mutation Detection Kit are cutting-edge tools that exemplify advancements in personalized medicine. These kits cater to oncologists and researchers aiming to refine therapeutic approaches and improve patient outcomes. Let’s explore the key features, benefits, and applications of these innovative diagnostic solutions.

The Importance of ROS1 Mutations in Cancer

ROS1 gene rearrangements are associated with several cancers, including non-small cell lung cancer (NSCLC). These mutations lead to abnormal protein fusion that drives tumorigenesis. Detecting ROS1 mutations is critical for identifying patients who may benefit from targeted therapies, such as ROS1 inhibitors.

The ROS1 PCR Kit offers a highly sensitive and specific method to identify ROS1 gene fusions. By utilizing advanced molecular techniques, the kit provides rapid and accurate results, making it indispensable for oncologists and researchers. This kit not only enhances diagnostic accuracy but also plays a significant role in the selection of precision therapies, thereby improving patient outcomes.

ROS1 PCR Kit: Key Features

High Sensitivity and Specificity: Detects ROS1 gene fusions with unparalleled precision.

Rapid Turnaround Time: Ensures timely diagnosis for initiating appropriate treatment plans.

User-Friendly Protocols: Designed for ease of use in clinical and research settings.

Comprehensive Coverage: Covers a wide range of ROS1 rearrangements to cater to diverse clinical needs.

By integrating the ROS1 Mutations Kit into diagnostic workflows, healthcare professionals can significantly enhance their ability to tailor treatments to individual patient profiles.

MYD88 Mutations: A Key Player in Hematologic Cancers

The MYD88 gene is implicated in various hematologic malignancies, including Waldenström’s Macroglobulinemia and diffuse large B-cell lymphoma (DLBCL). Mutations in MYD88, particularly the L265P mutation, are recognized as critical biomarkers for these conditions. Identifying these mutations can help in diagnostic stratification, prognosis determination, and therapy selection.

The MYD88 PCR Kit is a robust solution for detecting MYD88 L265P mutations. This kit leverages advanced PCR technology to deliver reliable and reproducible results, empowering clinicians and researchers to make informed decisions.

MYD88 Mutation Detection Kit: Key Features

Precision Targeting: Accurately identifies MYD88 L265P mutations.

Streamlined Workflow: Facilitates seamless integration into diagnostic protocols.

High Reproducibility: Ensures consistent results across multiple tests.

Versatile Applications: Suitable for both clinical and research environments.

Applications and Benefits

Both the ROS1 PCR Kit and MYD88 Mutation Detection Kit are integral to the paradigm of precision oncology. Here’s how they contribute:

Enhanced Diagnostic Accuracy: These kits reduce the risk of false positives or negatives, enabling precise identification of genetic alterations.

Personalized Treatment Plans: By detecting actionable mutations, these kits aid in tailoring therapies to individual patients.

Accelerated Research: Researchers can utilize these kits to uncover new insights into cancer biology and treatment.

Improved Patient Outcomes: Early and accurate detection translates to timely intervention and better prognoses.

Conclusion

The ROS1 and MYD88 Mutation Detection Kits are transformative tools in the realm of molecular diagnostics. By enabling precise detection of critical mutations, these kits pave the way for personalized medicine and improved cancer management. Whether in clinical diagnostics or cutting-edge research, these solutions underscore the importance of innovation in combating cancer.