Prospects for in vitro diagnosis (IVD)

In vitro diagnosis and the industry

In vitro diagnosis refers to products and services that obtain clinical diagnostic information by detecting human samples (blood, body fluids, tissues, etc.) outside the human body to determine disease or body function.

In vitro diagnostic products are mainly composed of diagnostic equipment (instrument) and diagnostic reagents. According to the National Food and Drug Administration (SFDA) “Medical Device Classification Catalogue” standard, in vitro diagnostic equipment belongs to the category of clinical laboratory analysis instruments.

The in vitro diagnostic industry and laboratory medicine constitute an organic whole that is both distinct and closely related. The in vitro diagnostic industry is the “tool” and “arms” of laboratory medicine, while the laboratory medicine is the “user” and “market” of the in vitro diagnostic industry. The common purpose of both is to perform in vitro diagnostics. About 80% of clinical diagnostic information comes from in vitro diagnosis, and its cost accounts for less than 20% of medical expenses. In vitro diagnosis has become an increasingly important part of human disease prevention, diagnosis and treatment, and an increasingly important part of safeguarding human health and building a harmonious society.

In vitro diagnostic reagents

In vitro diagnostic reagents refer to in vitro diagnostic reagents administered by medical devices, including alone or in combination with instruments, instruments, devices or systems, in disease prevention, diagnosis, treatment monitoring, prognosis observation, health status evaluation, and hereditary diseases. In the process of predicting, reagents, kits, calibrators (objects), and quality control products (objects) used for in vitro detection of human samples (various body fluids, cells, tissue samples, etc.).

Classification of in vitro diagnostic reagents

According to the level of product risk, in vitro diagnostic reagents are divided into Class I, Class II and Class III.

Class III: 1. Reagents related to the detection of pathogenic pathogen antigens, antibodies and nucleic acids; Reagents related to blood type and tissue matching; 3. Reagents related to human genetic testing; 4. Reagents associated with hereditary diseases; 5. Reagents related to the detection of narcotic drugs, psychotropic drugs, and medical toxic drugs; Reagents related to target detection of therapeutic drugs; 7. Reagents associated with tumor marker detection; Reagents associated with allergies (allergens).

Class II: in addition to the products that have been identified as the third category and the first category, the others are the second category of products, including: 1. Reagents for protein detection; 2. Reagents for sugar detection; 3. Reagents for hormone detection; 4. Reagents for enzyme detection; Reagents for ester detection; Reagents for vitamin testing; 7. Reagents for inorganic ion detection; Reagents for the detection of drugs and drug metabolites; Reagent for autoantibody detection; 10. Reagents for microbial identification or susceptibility testing; A reagent used for the detection of other physiological, biochemical or immunological indicators.

Class I: 1. Microbial culture medium (not used for microbial identification and susceptibility testing); 2. Sample processing products, such as hemolytic agents, diluents, staining solutions, etc.

 

Application prospects of in vitro diagnostics:

In vitro diagnostic technology is rapidly evolving, from gene-level gene sequencing, SNP screening, point mutation gene diagnosis, to protein levels of various biomarkers (eg, cancer biomarkers, breast cancer biomarkers, inflammatory biomarkers, lung cancer biomarkers, prostate cancer biomarkers, liver cancer biomarkers, nephropathy biomarkers, pancreatic cancer biomarkers) detection, to the cellular level of circulating tumor cell detection ( CTC), thin-layer liquid-based cytology (TCT), and PET/CT at the tissue level. In general, in vitro diagnostics are moving toward easier, faster, non-invasive, and more informative. The application prospects mainly include the following aspects:

 

  1. 1. Physical examination

Modern people are paying more and more attention to their own health. Many people have become a habit every year. In vitro diagnosis by detecting and testing blood, urine, feces, secretions, etc., disease prevention and early detection can be achieved.

  1. 2. Chronic disease management

Worldwide, the incidence of diabetes, high blood pressure, and chronic stomach diseases has increased year by year. The incidence of diseases such as hyperlipidemia and osteoporosis is also high. These chronic diseases require regular monitoring of changes in blood sugar, blood pressure, blood lipids, Helicobacter pylori, and bone calcium. The use of in vitro diagnostic equipment, especially various wearable devices, enables monitoring of blood glucose, blood pressure, heart rate, etc. anytime, anywhere.

  1. Critical illness monitoring

After many cancer patients undergo surgery and chemotherapy, doctors can use in vitro diagnostic techniques to regularly monitor tumor markers, predict cancer metastasis, and guide medication. For example, breast cancer patients should have regular chest X-ray, bone scan, liver B-ultrasound, blood routine, and tumor-associated antigen examination. Researchers at the British Columbia Cancer Research Center have found high expression of αB-crystallin gene in the brain through in vitro diagnostic techniques. Breast cancer patients who are positive (negative) face a three-fold or higher risk of brain metastasis of cancer cells than patients with low-expression (negative) breast cancer. This allows doctors to target patients at high risk to prevent cancer metastasis.

Introduction to the Amino Acid Modification

The amino acid substitution modification technique is a modification method in which one amino acid on a peptide chain of a protein molecule is replaced with another amino acid.

At a specific position of a protein molecule, the type and nature of the amino acid are specifically invariant. The amino acid substitution modification refers to changing a specific amino acid on the peptide chain to another amino acid, thereby changing the spatial configuration of the protein, thereby changing the enzyme. Certain biological properties of proteins, known as amino acid substitution modifications.

 

Enzyme protein amino acid substitution modification

The amino acid substitution modification mainly plays a role in increasing the enzyme activity of the enzyme protein and increasing the stability of the enzyme protein; for example, after the third isoleucine on the molecule of the T4-lysozyme enzyme protein is substituted with cysteine, Cysteine ​​can form a disulfide bond with the cysteine ​​at position 97, which plays an important role in maintaining the spatial configuration of the enzyme protein. The enzyme stability is increased by maintaining the enzyme activity. Times. Amino acid substitution modification methods are mainly carried out by means of genetic engineering.

 

Studies have shown that covalent binding of cofactors to enzymes can produce new enzyme activities. In recent years, more creative work has been to develop new coenzyme analogs and introduce them into the active site of the enzyme to achieve cofactor remodeling of the enzyme, which is to solve the problem of expensive cofactors in the catalytic reaction system. Important progress. Using the solid phase synthesis technique, the 8th residue phenylalanine in the c-peptide chain of ribonuclease s is replaced with a natural amino acid, pyridinium phosphate (vitamin B6), and the recombined enzyme catalyzes the reaction rate. Increased by 7 times.

 

Amino acid substitution modification methods

  1. Chemical modification method
    The most creative work of chemical modification is to make a catalyst with new catalytic properties by atomic displacement of the active site amino acid of the enzyme protein by a purely organic chemical reaction. The earliest example was the conversion of the serine active site of subtilisin to a cysteine ​​residue, resulting in the loss of its initial aminohydrolysis activity, but still retaining the ability to catalyze the esterification reaction. It can therefore be used for the synthesis of peptides. The selenocysteine ​​is introduced on the subtilisin to convert the aminohydrolase to an acyltransferase. They subsequently discovered that this selenium-based subtilisin has similar activity to glutathione peroxidase due to the presence of redox-active selenium atoms. Since then, other researchers have used this route for the modification of trypsin, which also has redox activity.

 

  1. Directed mutation technology
    Directed evolution has evolved into a fairly mature protein and peptide modification technology in its short span of more than a decade. Its application has greatly promoted the development of other related fields such as enzyme engineering, metabolic engineering and medicine. With the joint efforts of many researchers, the field of protein directed evolution has made outstanding achievements, and there have been many successful examples of directed evolution applied to industrial biocatalyst transformation.

 

Directed evolution can be applied not only to the significant improvement of biocatalyst activity, thermal stability and performance of the operating conditions, but also to the vaccine and pharmaceutical fields, to modify vaccines and protein drugs, and to achieve significant evolution in various applications. Directed evolution has provided a powerful tool for biocatalysts from laboratory research to industrial applications. At present, the gratifying results of some enzymes (or proteins), arsenate detoxification pathways, radiation resistance, biosynthetic pathways, enantioselectivity, antibody libraries, and directed site directed evolution have Encouraged scientists in many related fields.

 

About us

Creative Peptides is specialized in the process development and the manufacturing of bioactive peptides. We are dedicated to offering custom peptide synthesis, process development, GMP manufacturing as well as catalog products for customers in industry and research area. Here are some our products like: Peptide ModificationDe novo Peptide DesignLong Peptides SynthesisPeptide Drug Discovery, etc.

 

How to become a great medicinal chemist? (part two)

Mark A. Murcko combines his observations and experiences, summarizing the characteristics of many great medicinal chemists have in accordance with the two categories of “general” and “professional”, hoping to give aspirations to the field of medicinal chemistry.

  1. Managing the nature of the compound

A great medicinal chemist’s gaze is not limited to the efficacy of the drug, but rather to the overall characteristics of the drug. At the same time, they are not limited to various rules, through experimental data to improve the parameters of the compound, such as molecular weight, structural complexity, lipophilic and so on. If optimized in the right direction, active compounds that do not seem to be a drug may also surprise.

HCV encodes at least 10 mature proteins, and non-structural protein 5A (NS5A) is one of its important proteins, and is also an important target for anti-HCV drugs. The candidate compounds found by medicinal chemists are somewhat “anti-conventional”, with molecular weights of up to 750-900 and a logP of 8, which has made some pharmaceutical companies somewhat embarrassed. However, there are still some teams who are brave enough to face the challenge and successfully market their drugs, such as daclatasvir and ledipasvir, by optimizing the structure.

  1. Consider the three-dimensional structure

With the development of various technologies, drug designers can now easily obtain protein structures. But even if it is not, drug chemists have been contemplating the structure of compounds in three-dimensional space in their minds. How do they look, in water, and in lipids? Is there any difference between extracellular and intracellular? Does it change compared to the free state after binding to the target?

  1. Prepare “Plan B”

Unexpectedly, drug research and development is a systematic project, and problems in any part of pharmacokinetics, drug toxicity, compound synthesis, drug production, intellectual property, etc., will have serious consequences. There is a high-quality, new structure of alternative compounds that are undoubtedly the best contingency plan. For example, at the beginning of Relay Therapeutics, team members consciously set out to discover many different types of chemical structures for three reasons: to maximize access to molecules with good bioavailability; to protect their work more broadly; to verify that they are constructed a drug discovery platform. In the first year, Relay’s team discovered more than a dozen compounds with nanomolar bioactivity, enabling the team to quickly move to clinical development.

  1. Not worrying about intellectual property issues

Medicinal chemists must pay attention to patents. Sometimes, a number of companies aiming at “hot targets” in a bee-like project will lead to drug discovery around similar molecules, and patent homogenization competition is serious. But don’t forget that the “chemical space” is very large, and researchers are enough to jump out of existing patent restrictions and discover other drug molecules. There are sometimes many drug molecules targeting the same target, especially some drugs that follow the trend. Most of them have a common structure, but drug chemists can still retain their own characteristics and find unique “corners” in the “chemical space”. “By optimizing by target, each molecule has its own unique advantages.”

Disease Biomarkers for In Vitro Diagnostic (IVD) Development

In-Vitro Diagnostics (IVD) refers to the removal of blood, body fluids, tissues and other samples from the human body. The samples are tested and verified using in vitro detection reagents, kits, calibrators, and quality control materials. The process of prevention, diagnosis, treatment testing, post-observation, health evaluation, and genetic disease prediction.

Finding and discovering reliable biomarkers is critical for many common and major diseases. In recent years, with the development of immunology, molecular biology and genomics technology, scientists have achieved many important results. Relevant departments in various countries and regions have also introduced corresponding support policies, such as the Critical Path Initiative (CPI) issued by the US Food and Drug Administration (FDA) in 2004 and the European Medicines Agency Road Map to 2015 and the Organization for Economic Co-operation and Development (OECD) biomarkers in health applications and development policies clearly mention the key to biomarkers in new drug development and clinical treatment effect.

The development of basic and applied research on disease biomarkers, coupled with significant clinical needs and policy promotion, has also led to in vitro diagnostic(ivd) development, providing a powerful means for clinical testing.

  1. Progress in research oncancer biomarkers

1.1. Cancer biomarkers: researchers from the University of Würzburg, Germany, have used cancer molecules to diagnose cancer, and the study was published in the international journal ChemBioChem. Researcher Jurgen Seibel said that galectin-1 molecules help tumor cells hide. From the immune system attack, recent studies have suggested that when the function of galectin-1 is blocked, the immune cells will recognize the tumor and launch T cells to attack the tumor. Researcher Seibel and colleagues studied the specific part of the protein, the carbohydrate recognition domain, in which they designed a complex sugar molecule that would be well suited to the domain. They immobilized sugar molecules on a “parking site” and attached fluorescent dyes or drug molecules. In addition, the researchers described how the sugar molecule binds to galectin-1 with a high-resolution X-ray structure.

1.2. In a study published in the international journal Nature Medicine, researchers from Fudan University and Mayo Clinic in China identified a new mechanism of prostate cancer resistance to therapy through joint research. The development of new prostate cancer therapies provide ideas and hopes. In the article, the researchers explained that the mutations in the SPOP gene play a key role in the drug resistance of prostate cancer. The SPOP gene mutation is the most frequent genetic mutation in primary prostate cancer. These mutations are in the cancer pair. BET inhibitor drugs play an important role in the development of tolerance.

  1. Progress in research on cardiovascular biomarkers

Cardiovascular biomarkers: researchers from Michigan State University analyzed 240 patients in the emergency department to find out the relationship between cholesterol crystals and heart disease in patients. The study’s lead author, George Abela, chief cardiovascular specialist from MSU, analyzed the material composition of the heart arteries in patients with heart disease and found that 89% of them had excess cholesterol crystal deposits in the area.

  1. Progress in research on diabetes biomarkers

3.1. Special lymphocytes or new biomarkers that can be used as a form of type 1 diabetes. Researchers from the Necker Children’s Disease Hospital and the Cochin Institute have found that the onset of type 1 diabetes may be guided by the modification of MAIT lymphocytes, which recognize the components of the body’s microbial population. Thus as a novel biomarker for early diagnosis and prevention of diseases

3.2. The 3U Diabetes Alliance from Dublin City University, Eric West University and the Royal College of Surgeons of Ireland has a groundbreaking study that found biomarkers that might predict the development of type 1 diabetes. The researchers found that a substance called 12-HETE in the blood can be used to diagnose patients with type 1 diabetes. If the patient is not ill, the substance cannot be detected in the blood.

  1. Progress in research on biomarkers of nervous system diseases

4.1. Researchers from the Sanford Burnham Prebys Medical Discovery Institute identified a key peptide or helped early diagnosis of Alzheimer’s disease (AD), the results of which were published in the international journal Nature Communications. It can help researchers develop new ways to introduce drugs into the brain of patients to treat a variety of diseases, such as Alzheimer’s disease, Parkinson’s disease, stroke and glioblastoma.

4.2. In a study from Ohio State University, researchers seem to have found a diagnostic method that accurately predicts patients with Alzheimer’s disease. According to the author of the article, Mingjun Zhang, a professor of biomedical engineering at Ohio State University, first discovered biomarkers that could be accurately diagnosed: proteins that changed in the patient’s spinal fluid and blood. Specifically, as the disease of Alzheimer’s disease becomes severe, these proteins become longer, harder, and more susceptible to agglomeration.

Advantage

In vitro diagnostics provide significant clinical and economic benefits by providing targeted drug biomarkers to improve treatment efficiency while reducing complications. This approach can be based on the biomarker-based test results for drug development by correlating the efficacy of the drug with measurable biomarkers. In addition, this approach may increase the chances of success in key trials by centralizing the inclusion of patients with curative effects in clinical trials.

In recent years, the development of biomarker targeted therapies has been the focus of biomedical research. In 2016, 6 of the 22 new molecular entities (NMEs) and biologics approved by the FDA were targeted therapies, accounting for 27% of the total. Compared with the 14% in 2011 and the 5% in 2005, it can be seen that the approval rate of targeted therapeutics has increased significantly. Although the number of approved therapeutic drugs in the total number of approved NMEs continues to increase, it has been mainly concentrated in the field of cancer, and targeted therapy in other disease areas is very limited. There are many reasons for this phenomenon: first, even if there is some progress in the field of cancer treatment recently, there are still many unmet clinical needs in this field, especially the clinical efficacy is relatively poor, and the non-targeted treatment program is serious. Adverse events, etc. Second, our current scientific understanding of cancer and its genomic etiology is deeper than any other non-communicable disease. Therefore, many of the cellular mechanisms triggered by tumor-induced gene mutations can be targeted using appropriate drugs. Finally, the choice of patients with cancer treatment targets has a huge economic impact: cancer treatment is usually expensive and can significantly increase expenditure, but the economic benefits of targeted therapy for patients who may be effective are also obvious.

In general, biomarkers can be incorporated into clinical development programs in two ways. The first method is to use a genome-wide association study to determine the biomarker signature corresponding to a significant drug. Because this method is indirect (i.e., independent of any information related to known drug characteristics and mechanisms of action), it may be affected by a variety of factors (including biomarker distribution, population and geographic differences), so this method is less likely to bring value to the payer or the pharmaceutical company. The second method is more ideal, and the biomarkers are directly related to the method of action of the drug. This is a typical targeted tumor drug in which the biomarker itself may be a drug target. For example, epidermal growth factor receptor (EGFR) inhibitors (such as erlotinib) and human epidermal growth factor receptor 2 (HER2) inhibitors (such as pertuzumab). In other cases, biomarkers can identify another protein in the drug’s pathway of action, which can cause the drug to lose its effectiveness on the tumor (eg, RAS mutations can block the action of the HER2-targeted drug panitumumab). In any case, the correlation between biomarker detection and drug activity is first determined in a non-clinical (in vitro) study and then initially assessed in an early clinical study.

Any biomarker reagent for patient selection and enrollment in a stratified oncology trial will appear in the indication of the drug, so the reagent must be marketed as a companion diagnostic test available for sale. Companion diagnostics (CDx): means the use of in vitro testing equipment to provide certain disease information to help improve the safety and effectiveness of the corresponding drug during use. Concomitant diagnosis features: one is to screen patients who are suitable for treatment with specific drugs, and the other is to screen patients who are not suitable for specific drug treatment, that is, patients who may have serious side effects or have weak reactions to drugs, and thirdly, monitor the treatment effect of patients and guide the development of the most suitable treatment plan, and this is also the role of biomarkers. It helps to identify the population of patients most likely to respond to treatments, promotes the use of drugs in relatively limited markets, improves the effectiveness and safety of drugs, and reduces the waste of value due to weak patient response to drugs.

Biomarkers have become a key element in the current precise treatment or development of drugs. With the deeper understanding of the molecular mechanisms of disease, all disease treatment methods will eventually be able to develop biomarkers.

Reference

[1] Blackhall F, Peters S, Kerr K M, et al. Biomarkers[J]. Annals of Oncology, 2012, 23:ix73-ix94.

[2] Zhang Z, Jr B R, Yu Y, et al. Three biomarkers identified from serum proteomic analysis for the detection of early stage ovarian cancer[J]. Cancer Research, 2004, 64(16):5882.

[3] Li J, Zhang Z, Rosenzweig J, et al. Proteomics and bioinformatics approaches for identification of serum biomarkers to detect breast cancer.[J]. Clinical Chemistry, 2002, 48(8):1296.

[4] Zhang Z, Chan D W. The road from discovery to clinical diagnostics: lessons learned from the first FDA-cleared in vitro diagnostic multivariate index assay of proteomic biomarkers.[J]. Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology, 2010, 19(12):2995.

[5] Azzazy H M, Mansour M M. In vitro diagnostic prospects of nanoparticles[J]. Clinica Chimica Acta, 2009, 403(1):1-8.

[6] Lewandrowski K, Tamerius J, Menegus M, et al. Detection of Influenza A and B Viruses With the Sofia AnalyzerA Novel, Rapid Immunofluorescence-Based In Vitro Diagnostic Device[J]. American Journal of Clinical Pathology, 2013, 139(5):684-689.

[7] Listed N. Medical devices containing materials derived from animal sources (except for in vitro diagnostic devices), guidance for FDA reviewers and industry; availability–FDA. Notice[J]. Fed Regist, 1998, 63(215):60009-60010.

Biomarkers Promote Anti-tumor Drug Development

Biomarkers are the most direct and rapid diagnostic tools. Their screening and acquisition can play an important role in disease diagnosis, development, treatment, and efficacy monitoring. It is also an important target for drug development.

Biomarker refers to a biochemical indicator that can objectively measure and evaluate changes or possible changes in system, organ, tissue, cell and subcellular structure or function. It has a very wide range of uses and can be used for disease diagnosis and judgment. Disease staging or evaluation of the safety and efficacy of new drugs or new therapies in the target population can also help researchers to propose more effective treatments, especially in chronic diseases such as cancer, cardiovascular disease, diabetes, and neurological disorders. It has important value in the prevention and control of complex diseases.

Biomarkers in Cancer

  1. Nat Med: Chinese scientists discover new prostate cancer biomarkersthat are expected to improve individualized therapy for cancer

In a study published in the international journal Nature Medicine, researchers from Fudan University and Mayo Clinic in China identified a new mechanism of prostate cancer resistance to therapy through joint research. The development of new prostate cancer therapies provide ideas and hopes. In the article, the researchers explained that the mutations in the SPOP gene play a key role in the drug resistance of prostate cancer. The SPOP gene mutation is the most frequent genetic mutation in primary prostate cancer. These mutations are in the cancer pair. BET inhibitor drugs play an important role in the development of tolerance.

  1. Cell-metabolism:A team of scientists in Australia recently discovered a new type of lung cancer biomarker. These cancers have also been found to produce specific metabolites that may be identified in plasma samples, thereby increasing the hope that diagnostic blood test methods for such diseases can be developed in the future.

Kate Sutherland, co-director of the study, said, “One-fifth of the lung adenocarcinomas have changed in the KEAP1 / NRF2 pathway, indicating that it is a major cancer driver. These cancers are very aggressive and resistant to standard therapies. Force and poor prognosis, so new treatments are urgently needed.”

Up to 40% of lung cancers are lung adenocarcinomas, and one in five of these specific tumors are resistant to chemotherapy and radiation. The new study suggests that these specific cancers may respond well to a new generation of anti-PD-1 and anti-CTLA-4 immunotherapies. The ability to easily and quickly identify these specific tumors will enable physicians to better identify those who respond best to new immunotherapy treatments.

Another result of this new study was the discovery of unique metabolic markers that can be found in blood samples, pointing out potential early detection blood tests. “We worked with our colleague Dr. David De Souza and Prof. Malcolm McConville of Bio21 to identify a unique ‘breadcrumb’ trace of these cancers in the blood,” Sutherland said. “We hope that this test will identify patients who may respond to immunotherapy, and may also be a simple non-invasive blood test that can detect these lung cancers early.”

This study currently only finds this molecular pathway in animal models, so the next stage is to validate these results in human lung samples. Since then, clinical applications may take some time to become a reality, but this finding is critical to creating blood tests that both diagnose lung cancer and guide doctors to the most likely successful treatment.

  1. Nature Communications:Researchers from the National Cancer Research Center (CNIO) in Spain have successfully identified biomarkers for breast cancerthat can be classified for the first time. This is the first time that a curable patient can be distinguished from a patient who may relapse. Open. It also identified new pharmacological indicators and indicated that combination therapy with existing drugs may be effective in patients with these indicators.

Specifically, they identified six protein kinases whose functional status predicts the evolution of triple-negative breast cancer. In addition, researchers have found a way to detect these proteins in hospitals, so in the future it may develop into a routine clinical trial, just like the genetic analysis of any tumor today.

From genomics to proteomics

Research on cancer genomics for decades has revealed dominant gene mutations in many cancers that determine tumor progression and guide the design of individualized therapeutic approaches. This type of specific treatment for each type of tumor is more effective than traditional chemotherapy and is a major cause of advances in cancer treatment in recent years.

However, triple-negative breast cancer is caused by a variety of mutations that work together in a unique combination of each patient. To date, no dominant gene mutations have been identified that provide prognostic indicators or drug treatment responses.

To achieve this goal, CNIO researchers chose not to analyze the genes involved in triple-negative breast cancer, but to analyze their products: synthesize proteins sorted by these genes. Their hypothesis is that many of the patient’s genetic changes can be translated into an identifiable pattern of functional status of all tumor proteins and their proteomes – activation or not.

The experimental results were very successful. In a tumor sample of 34 patients, the researchers discovered biochemical markers of tumor protein activation. Although the initial number of candidates exceeds two million, with the help of sophisticated bioinformatics tools, they found that among all these signals, accurate combinations can only be found in patients who have relapsed. These proteins are activated by kinases, which in turn are proteins, so the next step is to find the kinase responsible for this signal. Finally, the study identified six kinases that led to the formation of a proliferating pattern of proteomes in relapsed patients.

Six kinases indicate possible recurrence: We currently know that these six kinases play a key role in triple negative breast cancer. In this study, the authors validated the results of 170 patients and confirmed the value of these six kinases as markers. Patients who do not have these proteins activated have a 95% chance of being cured, or at least not relapsed 12 years after treatment. However, even if one of the six kinases is active, the risk of recurrence increases tenfold.

New clinical trials targeting kinase activation: Analysis of the functional status of proteins is not currently routinely tested in hospitals, but the authors have translated the activation pattern of kinases into immunohistochemical indicators that can be easily analyzed in hospitals. The goal is to determine whether these six kinases can be routine clinical trials, just like the genetic profiling of any tumor today.

Currently, researchers are focusing on other markers of disease, including standardized kinase diagnostic tests, and clinical trials of patients with advanced disease using the therapeutic combinations described herein.

  1. Int J Cancer:Colorectal cancer biomarkers–application of fecal microbes in the detection of colorectal cancer.

Colorectal Cancer (CRC) is the second leading cause of cancer death in the Western world. CRC In most cases, genomic mutations, such as the APC/wnt signaling pathway, induce cell proliferation, which in turn leads to adenoma formation. The gradual accumulation of a driver mutation eventually leads to the formation of invasive tumors. If the CRC can be diagnosed at an early stage, the five-year survival rate can reach 80% or more. If the tumor cells have metastasized in the advanced stage, the five-year survival rate is less than 10%. Therefore, if early screening can be done in various countries, the mortality rate of CRC can be reduced.

The current fecal occult blood test (FOBT) is the most widely used colon cancer screening method, but its sensitivity and specificity for CRC detection is not high; colonoscopy is currently the most effective method for diagnosing CRC, but the operation is complicated and the patient experience is poor. Previous studies have found that changes in the intestinal flora are associated with CRC, but the potential of microbes as markers for colorectal cancer screening has not been clearly elucidated.

Therefore, researchers from Sweden used a nested case-control study to investigate the use of three microbial markers in fecal clBA + bacteria, afa-C+ diffuse E. coli, and Fusobacterium nucleatum in 238 subjects. The case of potential screening markers for CRC diagnosis.

The researchers found that clBA+ bacteria and individual markers of Fusobacterium nucleatum were more abundant in the stool of colorectal cancer patients and had higher specificity in predicting cancer (81.5% and 76.9%, respectively), and their sensitivity. They were 56.4% and 69.2%, respectively. In a combined trial of clBA+ bacteria and Fusobacterium nucleatum, the specificity of CRC diagnosis was 63.1% and the sensitivity was 84.6%.

The findings of this study support the ability of microbes in feces to serve as potential non-invasive biomarkers for CRC detection, and the method has potential value. The researchers suggest that, in the future, microbial markers may be an important screening strategy for patients with CRC who have “high risk” for other diagnostic methods such as colonoscopy.

Reference:

[1] Eklöf V, Löfgren-Burström A,et al.Cancer associated faecal microbial markers in colorectal cancer detection.Int J Cancer. 2017 Aug 22.

[2] Diamandis E P. Mass spectrometry as a diagnostic and a cancer biomarker discovery tool: opportunities and potential limitations.[J]. Molecular & Cellular Proteomics Mcp, 2004, 3(4):367.

[3] Sawyers C L. The cancer biomarker problem[J]. Nature, 2008, 452(7187):548-552.

[4] Zhang H D, Jiang L H, Sun D W, et al. CircRNA: a novel type of biomarker for cancer.[J]. Breast Cancer, 2018, 25(1):1-7.

[5] Costello E. A metabolomics-based biomarker signature discriminates pancreatic cancer from chronic pancreatitis[J]. Gut, 2018, 67(1):gutjnl-2016-313665.

How many kinds of signaling pathway do you know?

Signal pathway is the phenomenon that the signal transmits a kind of information from the outside of the cell to the inside of the cell when there is a certain reaction in the cell, according to which the cell has to react. The idea of Signal pathway dates back to 1972. But at that time, it was called signal transmission. The concept of signal transduction was widely used by M. Rodbell in a review in 1980.

Do you know Toll and Imd Signaling Pathway?

Inflammation induced by members of the TLR family (except TLR3) goes through a classic signaling pathway. This pathway originated from the Toll/IL-1 receptor homologous region (TIR), which is an intracellular conserved sequence of TLRs. TIR activates intracellular signaling mediators-interleukin-1 receptor-associated protein kinase (IL-1R associated kinase, IRAK) IRAK-1 and IRAK-4, TNFR-associated factor 6 (TRAF-6). Mitogen-activated protein kinase (mitogen activated protein kinase, MAPK and I κ B kinase, I κ K), NF-κ B) induces the expression of inflammatory factors.

 

The signal molecules involved in this signal transduction include:CD14,MD-2,TRAM,TRIF,TIRAP,MyD88,TLR1,TLR2,TLR3,TLR4,TLR5,TLR6,TLR7,TLR8,TLR9,IRAK-1,IRAK-2,IRAK-4,IRAK-M,TRAF6,TRIAD3A,ST2L,SOCS1,RIG-I,FADD,TOLLIP,RIP1,A20,UEV1A, Ubc13,ECSIT, MEKK-1, TAK1,TBK1, MKK3/6, p38, TAB1/2, MKK4/7, JNK, IKKα, IKKβ, IKKγ, IKKε,NEMO, IκBα, NF-κB, p65/RelA, Casp-8, IRF-3,IRF-7,MAVS and so on.

Do you know NOD-like Receptor Signaling Pathway?

By activating innate and adaptive immunity, microbes initiate a series of defensive processes. The congenital immune system consists of immune and non-immune cells expressed on the sensor or pattern recognition receptor (PRRS) and associated molecular patterns (PAMPS) from different pathogens. PAMPS recognizes inflammatory cytokines and type I interferons that trigger antimicrobial effects through induction. Toll like receptors, NOD receptors, RIG-I receptors and DNA sensors have been well studied. Let’s see what the signaling pathways of NOD receptors look like.

NOD like receptor is widely found in the cytoplasm of human cells whose corresponding ligand can activate NF-K B (NOD1 and NOD2) signaling pathway and activate Caspa se-1 to promote the production of IL-1B and IL-18, thus initiating innate and acquired immunity. NOD protein is the pattern recognition receptor in the plasma and its structure is as follows:

  1. 1.The central nucleotide binding oligodepolymerization region (NACHT) is a common structure of the NLRs family, which is very important for the oligomerization and activation of NLRs.
  2. 2.The N-terminal effect binding region is the domain of the interaction between N-terminal diWhite and protein.
  3. 3.The repeat sequence (LRRs), which is rich in leucine at the C-terminal, can recognize the receptor and the mutations of NOD2 and N-LRP3 in the NOD family can cause inflammation. So at present, the most studied is the NOD-like molecule. NOD1 and NOD2 can recognize different degradation products of bacterial peptidoglycan, and then activate two signal pathways, MAPK and NFBK, by RIP2 kinase (RICK), with the same CARD structure in its CARD oligoset.

Do you know RIG-I-like Receptor Signaling Pathway?

RIG-I is retinoic acid to induce gene protein I (RIG-I). RIG is a cellular receptor that recognizes viral double-stranded RNA and is a member of the RNA helicase family of DexD/H boxes. The C-terminal of RIG-I is a helical domain, which can be combined with synthetic double-stranded RNA and viral double-stranded RNA, and can be solved by ATP enzyme dependent way. The N-terminal is two series of cysteinase recruitment domain (CARD). RIG-I can recognize the RNA component of the virus and transmit signals by its own CARD interacting with the downstream signal molecule MAVS CARD. It activates the transcription factors IRF-3 and NF-κB into the nucleus and induces the expression of interferon B. In order to initiate the innate immune response and regulate the subsequent acquired immune response, it can enhance the ability of the body to resist the virus. In addition, in recent years, two viral RNA recognition proteins, MDA5 and LGP2, have been found to be similar in sequence and function to RIG-I too.

Monoclonal Antibodies—a Valuable Asset in Human History (Part Two)

Abstract: Monoclonal antibody (McAb) was produced by B cell hybridoma cell line. Single antibody monoclonal antibody technology targeting only one antigenic determinant was first initiated in 1975 by Koehler of Germany and Milstein of the United Kingdom. The basic principle of McAb technology is that mouse myeloma cells can proliferate indefinitely and secrete immunoglobulins without antibody activity in vitro and in vivo, while spleen cells of immunized mice have the ability to produce antibodies, but cannot proliferate indefinitely. These two cells are fused into hybridoma cells using a fusion agent (PEG, etc.). Such hybridoma cells have the major features of the parental cells. It can not only proliferate in artificial culture, but also produce specific antibodies. Clones grown from a single cell can be selected by limiting dilution. McAb has many advantages, such as: high unity and homogeneity; high titer; immune antigen does not need to be purified, but pure antibodies can be obtained; different specificities (groups, types, strains) can be obtained; high yield, continuous production, etc. Among these advantages, especially the uniform high specificity, McAb has a very wide practical value, and has formed a very popular emerging industry in the world. Monoclonal antibody technology based on its simplicity Easy, fast, specific, and sensitive Ubiquitous application for disease diagnosis and treatment, food hygiene, etc. The field has broad application prospects.

Keywords: monoclonal antibody, preparation, application, prospects

The development prospect of monoclonal antibodies

Fully humanized mAb

Since the advent of hybridoma technology, monoclonal antibodies have been widely used. However, most of the monoclonal antibodies are of murine origin, and anti-mouse antibodies are produced in vivo during repeated clinical administration, so that the clinical efficacy is weakened or disappeared. Therefore, the ideal monoclonal antibody for clinical application should be of human origin. Therefore, the genes for controlling spleen production in mouse fertilized eggs can be cut off by genetic engineering and gene processing technology, and the human spleen-producing genes in human cells can be extracted and transferred into mouse fertilized eggs to be fertilized. After adult mice, antibodies produced by the spleen in mice are fully humanized antibodies. This technology has good development and application prospects.

High targeting

Monoclonal antibody drug therapy mainly uses its targeting to intervene in various pathways of the development and progression of target cells, or to activate the host’s immunity to tumors. Therefore, the target is the key to study different kinds of antibody drugs. With the continuous development of biomedicine, there will be a more targeted monoclonal antibody and a more powerful “warhead”.

Clinical diagnosis and detection

Monoclonal antibodies are superior to existing antisera in the diagnosis of infectious diseases, immune diseases, endocrine diseases and early pregnancy diagnosis. For example, AIDS has become a major “killer” for human beings. It spreads very quickly and has a certain incubation period in the human body. The patient has no response, which delays the optimal treatment period. For example, HIV is extracted to make a monoclonal antibody, and the monoclonal antibody is made into a card of the same size as the pH test paper. So just put a small card in your mouth and you can detect if you have AIDS. This is a good way to prevent and detect AIDS and to enable patients to receive treatment earlier.

Lifetime immunity

The monoclonal antibody against a certain antigen is introduced into the DNA fragment which controls the synthesis of the corresponding monoclonal antibody by protein engineering technology, and then the corresponding DNA fragment is transferred into the human body by gene therapy technology, but the DNA fragment can be expressed in the body cell and replicated in the human body. Lifetime contains the corresponding antibodies against the corresponding pathogens.

Chimeric antibodies and humanized antibodies are also promising in the field of genetic engineering. If you are interested, please check them out!

References

[1] Hon-Ming Lam, Xun Xu, Xin Liu et al. Resequencng of 31 wild and cultivated soybean genomes identifies patterns of genetic diversity and selection [J]. Nature Genetics, 2010, 42(1038): 1053-1059

[2] Qingyou Xia, Yiran Guo, Ze Zhang et al. Complete Resequening of 40 Genomes Reveals Domestication Enents and Genes in Silkworm [J]. Science, 2009, 326(5951): 433-436

Math can predict how cancer cells evolve

Applied mathematics can be a powerful tool in helping predict the genesis and evolution of different types of cancers, a study from the University of Waterloo has found.

The study used a form of mathematical analysis called evolutionary dynamics to look at how malignant mutations evolve in both stem and non-stem cells in colorectal and intestinal cancers.

“Using applied math to map out the evolution of cancer has the potential to give oncologists a kind of road map to track the progression of a particular cancer and essentially captures crucial details of the evolution of the disease.” said Mohammad Kohandel, an associate professor of applied mathematics at Waterloo. “Combining the use of applied math with previous research advances in cancer biology, can contribute to a much deeper understanding of this disease on several fronts.”

The study found when cancer stem cells divide and replicate, the new cells that are created can be substantially different from the original cell. This characteristic can have a substantial impact on the progression of cancer in both positive and negative ways and the use of math can help better predict cell behaviour.

The study also concluded that this type of analysis may be useful in preventing the emergence of cancer cells, in addition to helping develop more intense and effective treatments.

“Being able to predict the evolution of cancer cells could be crucial to tailoring treatments that will target them effectively,” said Siv Sivaloganathan, a professor and chair of the department of applied mathematics, at Waterloo. “It may also help avoid the drug-induced resistance known to develop in many cancers.

“In addition to predicting the behaviour of cancer cells, this mathematical framework can also be applied more generally to other areas, including population genetics and ecology.”

see more at: https://www.cd-genomics.com/Bioinformatics-Services.html

How to Purchase Quality Pet Supplies

The trend in pet supplies, especially pet food, as of 2011 is organic pet food. While in the earlier days, organic food was carried only by health food stores, today even the local grocery stores carry them. Although more expensive than regular pet food, their advantages outweigh their price.

Pet Supplies and Accessories

Organic pet supplies now span even toys, pet beds and grooming products. For instance, you can now find organic cat litter, created from natural and recycled materials. These are good for the environment as well as for the cat. Similarly, there are environment-friendly doghouses that are UV protected, weather treated and ventilated well for your pet’s comfort.

Other poplar supplies include shampoos, flea medications, constraint cable, dog dryers and de-matting brush. While dryers remove water easily out of the dog’s coat, a de-matting brush, compared to an average pet comb or brush, makes grooming easier.

How to Find Quality Pet Supplies Wholesale

Here are some tips to find the best pet supplies:

    • Always buy products from companies that offer few, but high quality products. This is because small companies usually focus on customer satisfaction to retain a steady flow of customers.

 

    • Start shopping only after making a list of the supplies you need. You can also read customer reviews to take a decision.

 

    • Another key thing to consider is whether your pet will enjoy the things you buy. While earlier, a bone was a good enough toy, pet owners today splurge on all kinds of pet products. For instance, they largely splurge on fashionable clothing and sweaters. However, pet owners must remember that dogs do not need these, especially if they live in a tropical climate. In fact, these might harm your pet than do any good. This, along with the combined effect of fur overheats their bodies, leading to health problems. So, always consult a vet before purchasing clothing for our dog.

 

    • Ensure that you never spend more on your pet’s clothing than on its health care needs. Your dog’s health should always be your top priority. So, budget your pet expenses well.

 

  • As pet foods grow in popularity, so does its price. So, never fall for the temptation of buying the most advertised pet food. Instead, you must check the nutritional facts given on the pack to check the food’s worth. Veterinarians suggest that you replace packaged pet foods with organic food that you can cook at home. So, consult a vet before planning your pet’s new diet.

 

Wholesale Wedding Shoes Could Be the Answer to Finding Your Bridesmaid Shoes

If you are getting married this summer and have yet to buy shoes for your bridesmaids then why not look into buying a box of whole sale shoes. Buying your bridesmaids shoes off the high street can end up costing you a small fortune and you may struggle to find one pair that is available in everyone’s sizes, this could mean that you need to choose a different shoe or that you need to visit different stores to collect shoes which takes up time that you need to spend finalising your wedding details.

When you purchase wholesale shoes you will be able to purchase one box which has pairs in all sizes so you can be assured that all of your bridesmaids will be wearing the same shoe and no one will look out of place in a set of heels different to the others.

Most wholesale shoes come in boxes of 12 so any pairs that you do not use for your bridesmaids could be sold on an online auction website giving someone else the opportunity to find shoes they like in the sizes they need for their bridesmaids. The shoe sizes range from a 3 to 8 with the most popular sizes having multiple pairs, so you will hopefully have enough pairs in the right sizes for your bridesmaids.

There are many different styles of shoes available for your bridesmaids, you can go for a shoe with a low heel which they should be comfortable wearing all day or you may wish to choose two pairs shoes which will look fabulous on the photographs and shoes which they will be comfortable wearing all day. Don’t ever feel limited to the shoes available in the bridal section, always be sure to take a look at all other shoes as you may just find your perfect shoe for your bridesmaids hidden amongst the sandals. Most of the standard shoes will be available in a variety of colours so finding one to suit the colour of your bridesmaid’s dresses shouldn’t be difficult, the only difficulty you may have is getting all of your bridesmaids to decide on one shoe.

If you can’t find a suitable shoe for yourself then you may want to visit a wholesale shoe website because you can always sell the other bridal shoes in the box you purchase for your own shoes. There are bridal shoes with a low kitten heel which you should be comfortable wearing all day, or you may prefer to choose a higher heel if you are used to wearing them as these may be more comfortable for you.

Many brides and bridesmaids now have two dresses one for the ceremony and one for the reception; if you are having two dresses then you may find it beneficial to have two pairs of shoes. You should look for a pair of shoes which will be comfortable for you to wear during the ceremony and wedding breakfast and then a separate pair of shoes for dancing at the reception.

You should not feel restricted to shopping in bridal stores for your shoes, you need a pair shoes which you like and which you are comfortable in so if that means shopping online, wholesale or on the high street then that is what you should do. Just make sure that you have your shoes enough time before any alterations are made so that your dress is the correct length for the shoes you will be wearing. Your shoes need to be right for you if that means wearing gladiator sandals or stiletto heels, you need to be comfortable on your special day.

Article Source: https://www.import-express.com/apa/shoes.html